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2202true 江洪,吴蔚 2201true 孙玲 2200true 刘平,蒋冰冰,张礼邦,唐民圭,洪嘉禾,阮望春,刘成,俞广声,徐列明,薛惠明,赵佩珠,王玉润 2199true 宋文霞,王锦兰 2198true 王嘉玺,孙苏华,郅玉宝 2197true 曹锡清,毕蕾,潘华珍 2196true 赵宝贞,文德成,黄芬 2195true 于志洁,王宗伍,韩玲,陈铁河,潘予沙,苏福强 2194true 李宝珪,吴大庆,仲金良,黄道培,曾庆镒,卢惠卿 2193true 董伟,李新建,陶国枢,张玲 2192true 翁其亮,万田郎,苏祖佑,曹容珍 2191true 张淑辉 2190true 傅乃武,全兰萍,叶树勇,张谨,司德义,赵淑芳,贾德强 2189true 陈永青 2188true 关连 2187true 李美芬 2186true 吴同乐,张鸿卿,宋平根,袁传照,张成 2185true 周启玲,邢国仁,郑德存,柳青梅,陈去恶 2184true 陈敏,徐迎辉,许华林 2183true 潘华珍,李彩华,靳艳 2182true 沈霞昌,石彪,H.Ti Tien 2181true 李益新,董元林 2180true 赵孟莲,牛敏英,方家椿,薛绍白,宋平根,汪堃仁 2179true 苏兆众,罗祖玉,胡蓓华 2178true 金长清 2177true 肖奕,陈润生 2176true 3变种的作用，观察到硒砷的毒性可相互抑制，其相互作用由于浓度因素的影响既可表现出协同效应也可表现为拮抗作用。硒砷两微量元素共存也存在类似Bertrand关于单个元素的最适营养浓度。此外，还进一步分析了硒砷相互作用可能存在的途径。]]> 周召银,苏嫦,孙杰 2175true 王维光,顾俭本 2174true 刘孝全,刘心明 2173true 615(原发性)和H22(OAAT诱发)肝癌组织中不依赖于cAMP的蛋白激酶进行了研究，发现当以内源性蛋白为底物时，H615和H22组织中的酶活力分别是正常肝脏的3.10和2.44倍。以脱磷卵黄高磷蛋白和鱼精蛋白为底物时，肿瘤组织的酶活力是正常肝脏的1.5—4.45倍。研究结果表明，不依赖于cAMP的蛋白激酶活力升高与癌变和癌恶性状态的维持有密切联系。]]> 林晞,孙芝琳,刘宗定,蓝天鹤 2172true 周慧,赵志壮,付学奇,林永齐 2171true 姚松年,边书民,石智捷,李美亚,曹连欣 2170true 王明锁,孙国器,蔡伟波,冯纪辛,易剑 2169true 沈持衡,欧阳培 2168true 李钦,汪宁梅 2167true -1，t1／2等于4.53min，推测DMAP的透过方式为易化扩散。]]> 叶玲,黄如衡 2166true 沈孟长,陈晓青,罗勤慧 2165true 杨文学,易禄康 2164true 王立乾 2163true 许彩民,靳艳,吴元德,潘华珍 2162true +梯度时底物仍可使内膜流动性增加，提示流动性的增加与底物氧化启动的电子沿呼吸链的传递过程密切相关。]]> 冯元怡,程伯基,林克椿 2161true M可致红细胞溶血，10～40μM可促进K+外逸。1—2%牛血清白蛋白可以对抗这些效应．5—10μM棉酚明显抑制红细胞膜(Na++K+)-ATP酶活性，Ki值6.0μM。]]> 符云峰,张世联,王薇 2160true 任家鸣,赵天睿,南国华 2159true 2胆固醇含量和肾上腺维生素C含量的作用。而150mT磁处理水上述三种升高作用更为显著，同时还有降低血清总胆固醇和HDL3胆固醇含量的作用。两种磁处理水都具有防止主动脉内皮细胞损伤的效应，其中150mT磁处理水的这种预防作用更为显著。]]> 潘文干,张万超,孙奎,罗卫国,吴德林,康力 2158true 苏兆众 2157true 3H]DNA的方法。用该方法制备λ[ 3H]DNA，每升培养物可得10mg以上的产物。比放射性为6.75×104cpm／μGλDNA，酸不溶的放射性产物达99%以上。用单链和天然的λ[ 3H]DNA作底物，能方便地检测出酶制剂中极微量的核酸外切酶Ⅰ与Ⅱ和Ⅲ的污染活性。]]> 刘佑国,刘中昌,宋桂云 2156true CV值增大。(4)EB为10—20μg/ml、PI为50μg/ml是最佳荧光染色浓度。]]> 左连富,刘洪祥,郭建文,林元珠 2155true 王建一,阎新文,张苏玲,张天泽 2154true +离子传感器,当膜两侧溶液存在着质子梯度时,念珠菌素起着一个钠离子载体的作用。]]> 邰子厚,H.Ti.Tien 2153true 王少华,范秀娣,鲁瑜 2152true 安德荣,魏宁生,R.Koenig 2151true 黄耀煊,程志英,邬光惠 2150true Dictyostelium discoideum)基因组克隆的核苷酸顺序。这段顺序编码259个氨基酸,并含有一基因5’端非编码区结构。非编码区结构特点及氨基酸顺序表明这是一个新的肌动蛋白基因。基因组中约有15个内切酶消化所得的片段含有与此克隆同源的顺序。]]> 周世宁 2149true 林植芳,王伟,林桂珠,陈绵达,赵成学 2148true 李继珩,翁元凯 2147true 曹凯鸣,李碧羽,詹树萱,孙崇荣 2146true 35S-α-dATP为标记化合物,经缺口转移体外标记探针DNA,用旋转柱层析法分离出标记探针,由膜上DNA-DNA杂交,将ArNPV的多角体蛋白基因定位,确定是在其EcoRⅠ-Ⅱ片段上。]]> 陈蔚梅,林栖凤,马延高,卢文钩 2145true 杨东丽,崔耀宗 2144true 杨苍珍,马金石,蒋丽金 2143true 王厚行,李青 2142true 王昌才,陈仕荣,吴承声,金福军,钟雄林 2141true 鲜学义,韩相田,关鲁波,庞诚 2140true 2-·)处理分离的红细胞膜,研究外源自由基对红细胞膜的过氧化作用。结果表明,邻苯三酚对红细胞膜结构和功能有明显的损伤作用,导致膜流动性下降,膜荧光物质增加,膜蛋白发生交联作用以及膜重新封闭能力下降。]]> 秦德安,何学民,王永江 2139true 魏重琴,艾建芳 2138true 臧伟庆 2137true 秦安慰,苏兆众,何曼君,王玮 2136true 10后的代谢变化。结果表明,线粒体中CoQ同系物总量约占心肌全匀浆的19.49％;外源性的CoQ10能够为线粒体所摄取。]]> 范崇竹,张殿清,于维汉,于波,张洪印,王毓梅 2135true KSM)和链霉素抗血清对二链霉胺的亲和力(KB)之比(KSM/KB)为指标,表征链霉素抗血清抗原结合部位的特异性;建立了快速分析该指标的ELISA(酶联免疫吸附法)法;并对链霉素抗体抗原结合部位的特异性进行了分类。]]> 胡昌勤,赵建西,金少鸿 2134true P＜0.05);同种作物不同品种的种子在蔗糖溶液中萌发时发光强度表现出差异;用蒸馏水和蔗糖分别处理不同抗旱性的品种,则呈现与抗性有关的发光现象。]]> 杨起简 2133true 60的生长及DNA合成均有剂量依赖的抑制作用。NBT反应证明DFMO能诱导HL60细胞分化。核酸分子杂交实验表明,DFMO引起c-myc基因表达的减少,而c-fos基因表达水平随DFMO浓度增加明显增加。c-myc与c-fos表达的相互变化可能与DFMO对HL60细胞生长、分化影响有一定关系。]]> 范慕贞,齐上乐,赵成龙,马宇玲,吴克斌 2132true 陈学民,陈开浚,李孝光 2131true 邰子厚,余宝元,朱德煦 2130true 刘敬忠,T.HARANA,T.H.J.HUISMAN 2129true 路雪雅 2128true 王敦瑞,陈季,汪肖钢,于丽莉,陈诗书 2127true 傅乃武,黄磊,许慧君 2126true 段有金,李立,马萍,张仲林,李秀英,李芳生 2125true 刘道鸣 2124true 区耀华,吕冬,周昕 2123true 鲍仕登,汪德耀 2122true 刘军,梁明理 2121true 唐建民,傅昌余,王礼兵,刘代玺,陈祖林 2120true 2-·自由基(碱性条件下,邻苯三酚自氧化产生)处理后有明显损伤作用。在此基础上,本文研究猪红细胞SOD对人红细胞膜氧化损伤的保护作用。结果表明,外源SOD对人红细胞膜有明显保护作用,表现为膜流动性和膜重封闭能力保持正常水平,膜蛋白交联作用有显著减小。]]> 秦德安,何学民,王永江 2119true 180、P388、H22)也具有杀伤作用。在胰RNA提取物中,对瘤细胞具有杀伤作用的活性物质不是外源性凝集素样多糖类或能透过半透膜的低分子化合物,而是具有一定二级结构的低分子RNA。]]> 郑益星,韩明爱,申忠健 2118true 125Ⅰ-胰RNA能参入体外培养的瘤细咆中抑制DNA合成因而导致瘤细胞死亡。在以上实验基础上,我们又在动物体内注射胰RNA,观察对三种(S180、EC、P388)带瘤小鼠的影响,其结果:(1)抑瘤率为68.3％。(2)连续观察90天,给药组与对照组比较,两组的平均存活日数差异非常显著(P＜0.01),生命延长率为372％。(3)T.B淋巴细胞百分率及病理组织学变化,给药组与对照组比较都有明显差异。]]> 韩明爱,郑益星,崔京子,宋顺根 2117true 2电极的硅橡胶气透膜上制成的赖氨酸脱羧酶酶电极,性能如下:(1)酶电极对赖氨酸的线性响应浓度范围为0.0025—0.1％;极差为50—55mV;CV值小于5％;(2)连续使用寿命超过30天;(3)高度专一;(4)用于测定发酵过程赖氨酸浓度变化,结果与瓦氏呼吸仪的数据平行,相关性良好。]]> 吕东津,周润琦,陈石根 2116true 罗广祥,张丽青,顾梓伟,吉永华,徐科 2115true 60Co照射后,用HPLC结合AAS和ELISA方法检测第1,5,15,27天小鼠肝脏中金属硫蛋白两种亚型——MT-Ⅰ和MT-Ⅱ的变化发现,照后第1,5天MT-Ⅰ增加10—20倍,MT-Ⅱ增加40—60倍;而且600Rad照射较400Rad照射增加更明显,第15天后趋于正常。]]> 潘爱华,张瑞钧,茹炳根 2114true 60Co in order to study the effects of radiation on superoxide anion radical (O2-·) generation and the inhibitory effects of α2-macroglobulin (α2M) blood preparation. It was found that one hour after irradiation, the amount of O2-· released from polymorphonuclear leukocytes (PMNs) was more than that of the non-irradiated group (P＜0.01) and the activities of superoxide dismutase (SOD) in red cells decreased (P＜0.01). When α2M (138.5 U/ml blood) were added one hour before irradiation, the amount of O2-· released from PMNs were reduced and the activities of SOD in red cells were preserved. These results suggest that one reason of α2M against radiation may be related to inhibition of O2-· released from PMNs.]]> Liu Li and Jin Weiqiao 2113true 1O2,H2O2, O2-· and so on. The higher the concentraction of Ca2+-channel blocker——Verapamil presented, the lower the intensity of human Ly-CL observed. Probably, human Ly-CL depended on increased intracellular [Ca2+] of lymphocyte stimulated with antigens or mitogens. These findings propose that human Ly-CL may be an early events of lymphocyte activation.]]> Zhang Xuejun,Liu Lian and Si Chanping 2112true Xu Youhai 2111true 200, P300 could be related to the increased metabolism rate of brain energy and the decreased biosynthesis of bioactive substances.]]> Shen Zheng,Lin Shuzhi and Xie Jianong 2110true 125I and its specific radioactivity was 8.6μci/μg. We have utilized the autoradiography combined with scanning electro microscope(SEM-ARG) to observe the lectin receptors on liver cells, kidney ceils and rabbit red cells. The developed silver grains could be seen clearly on these cell surfaces. The silver grains represent the lectin receptor sites on cell surfaces. We found that the grains on liver cells were more than those on cells from other tissues. This result is consistent with the result which has been performed by the experiment of the specificity of chicken lactose lectin. Therefore,it is suggested that the chicken embryo liver lectin has certain specificity.]]> Chen Qingxuan and Shi Yingxian 2109true s can be self-assembled on a freshly cleaved metal wire by a twostep procedure: 1. The tip of a Teflon-coated platinum wire, while immersed in a lipid solution, is cut off with a scalpel; 2. The new tip of the wire, having become coated with lipid solution, is transferred into 0.1 mol/l KCl. After a few minutes, a stable lipid bilayer forms spontaneously on the tip of wire, as verified by electrical measurements. This paper reports an application of such a supported BLM (S-BLM) in the detection of Pb2+ ions. The S-BLM is liquidcrystalline in structure, which makes it amenable to modification for basic studies, as well as for technological applications such as biobensors and molecular electronic devices.]]> H. Ti TIEN and Z. SALAMON 2108true Hu Kunsheng,Wang Dahui and Tan Manqi 2107true Guo Zuo,Chen Yizhang,Chen Qi and Xu Renbao 2106true p＜0.01). The positive rate was 90%. In contrast serum GST-π in 25 cases of chronic hepatitis was determined to be 1.74 + 1.16ng/ml, which was not significantly different from the normal value (p＞0.05). The pseudo-positive rate was 12.0%.]]> Lin Feng and Chen Hui-li 2105true 2L2Cl2 with S-methyldithiocarbazate ligand was synthesized and characterized by various physical methods. It has been found that the complex has an activity of inhibition against some moulds bacteria and yeasts. Redox reactions on bilayer lipid membrane (BLM) modified by Cu2L2Cl2 was investigated by cyclic voltammetry method and the action of electron carrier of this complex on BLM was reported.]]> Yu Baoyuan,Tai Zihou and Sun Bingyin 2104true 7AM/ml proved the fit concentration for trapping the signal of the hydroxyl radical from AM in this experiment. 4. There was a great influence of DETAPAC or EDTA on the intension of the signal.]]> Hu Lianping,Sun Cunpu,Cong Jianbo,Gong Yifen,Wu Ke and Wu Dechang 2103true 6 and rutin. The dissociation constants of medicaments with albumin were obtained. The distances between the number 214 tryptophane and binding sites of vitamin B6 and rutin on the protein are 23.4Å and 24.02Å respectively.]]> Ma Guibin and Yang Pin 2102true Liu Jingzhong and Tang Qizhi 2101true 3 content. GM3 isolated from compacted canine erythrocytes with a yield of 351.0μg per ml and the purity was 92.2％.]]> Tsui Zhaochun,Hou Weihong and Zhu Zhengmei 2100true 31P-NMR to study the high energy phosphates of mouse L7811 ascites cells with thymic ceils from 615 line mouse as control. The results showed there was clear differences between them.Even in the late stage of growth, L7811 ascites cells was not completely in an inactive state. For example, the ratio of phosphocreatine (PCr) and α-, β-, γ- adenosine triphosphate (α-, β-, γ-ATP) to phosphate (Pi) was greatly decreased but not completely inactive. The ratio of sugar phosphate (Sp) to phosphocholine (Pcho) plus phosphoethanolamine (PEtn) increased, while glycero(3) phosphocholine (GroPCho) plus glycero(3) phosphoelhano-lamine (Gro PEtn) decreased. Preliminary studies showed that 31P-NMR spectra are sensitive monitor to the progressive changes of the high energy phosphate metabolism of the L7811 ascites growth.]]> Zhang Naizhong,Wang Fengru,Ma Liying,Han Qinhong,Wang Junheng,Liu Xiuping,Wu Jiazhen and Zhou Shuhua 2099true 2+, no change by sodium channel blocker——Tetrodotoxin, but abolished by calcium channel blocker——Cd2+. [65Zn2+]i was decreased by Cd2+. It is possible, that Zn2+, instead of Ca2+, goes into cells through calcium channels in the mechanism of BD evoked by Zn2+.]]> Wu Benjie and Zhang Chuanli 2098true Ding Yan,Chai Zhenming and Chen Chuanjuan 2097true 2/g of resin, the attaching yields of peptide to macroporous resin were higher than that of the gel type of resin. Macroporous TEPA resin has been demonstrated to be qualified as a polymer support for solid-phase sequence analysis of peptide.]]> Guo Xiaoli,Liang Xun and He Binglin 2096true Ostrinia furnacalis(Guenèe). Two strains were insecticide-resistant and one was susceptible. It is suggested that there were distinct differences in the zymograms of the esterase isozymes among the three strains, especially carboxylesterase. The number of cartoxylesterase bands of both resistant strains was more than that of the susceptible one, but there was no difference in thir cholinesterase. The zymograms were taken as a criterion to differentiate the resistant strain from the susceptible one. The method is suitable for studying resistance of some other agricultural insects and some aspects of methodology were discussed.]]> Shang Zhizhen and Xu Wenna 2095true Cao Enhua,Xin Shumin,Wu Wei,Cao Shaoqi,Zhang Youling and Lin Bohai 2094true Zeng Yitao,Huang Shuzhen,Zhou Xiadi,Zhu Hao,Chen Meijue,Li Houjun,Li Huiwu,Li Li,Zhao Xianning,Xing Fali,Chang Li,Jiao Chuntang,Tang Zhigui and Chen Chongyuan 2093true 32P-dCTP. The results showed that the PvuⅡ and HindⅢ mappings of three cases among the seven had been remarkably altered and the multiple changes of PvuⅡ sites had been observed, as compared with the normal subjuct. Therefore, the authors predicted that Ph chromasomal rearrangement in this kind of chronic myeloid leukemia could involve the inside of c-abl oncogene and might be a multiple rearrangement as well.]]> Li Hong,Wu Jinxu and Sun Zhilin 2092true 2SO4) ionic strength condition. The inhibited and residual transcription in high-salt system and presence of α-amanitin represented the activities engaged by RNA Pol II and Poi III independently, ihe transcription in low-salt system containing α-amanitin was mainly responsible for RNA Pol I activities. In addition, some factors effecting on the nuclei transcription in vitro were also evaluated. It was concluded that the reported model had advantages being easy to operated, saving materials, and approching cell physiological conditions, it was suitable for studying transcriptional mechanism of cerebral cortex neurons.]]> Qiu Ping and Yi Lukang 2091true +mRNA by Oligo (dT)-cellulose chromatography were performed. The difference of Poly(A)+mRNA between control and heat-shocked rat liver was shown by urea agarose gel electrophoresis. In vitro translation of Poly(A)+mRNA in wheat germ system, the synthesis of four major heat shock polypeptides with approximate molecular weights of 71000, 90000, 98000 and 110000 Dalton was observed. The relative amount of major heat shock proteins inereased as the rectal temperature of the heat-exposed rats rose. These results indicated that HSP was atso induced when rats, as a whole, were exposed to a simulated hot enviroment.]]> Chen Yuqing,Yan Guozhen,Chen Xiong and Cheng Suqi 2090true LIN Zhifang 2089true E. coli alkaline phosphotase gene from pFOG405 plasmid has been disclosed and compared with the sequence reported by Kikuchi et al. The result shows that the PvuⅡ-HpaⅡ fragment (144bp) containing the promoter and signal peptide sequence from pYK190 plasmid is sufficient for expression and secretion of foreign genes in E. coli. The deletion of the dyad sequence (26bp) from 5' end of the promoter region doesn't affect expression level of foreign target genes. The experiment also provids the useful information for construction of efficient expression and secretion vector using the PvuⅡ-HpaⅡ fragment and studying regulation of gene expression.]]> Jing Guozhong,Liu Lijun and Jiang Meiyan 2088true 65) was coupled to ricin using three different cross-agents respectively. Immunotoxins (ITs) with greater cytotoxic potency were obtained through a proper scheme. The ITs reduced the 3H-leucine incorporation of Molt-4 cells by 50％ at a concentration (IC50) of 2.7×10-12mol/L. ITs prepared with disulfide bond is 5—10 fold more cytotoxic in vitro than those prepared with thioether bond. The speed of metabolism of the latter in vivo is two times slower than that of the former. Because these two kinds of ITs have different degradation products, their metabolism pathways in vivo may be different also.]]> Ji Yanshan,He Yonghuai,Chen Xing,Zhao Weiwei and Shen Beifen 2087true Zhang Chunling,Zhang Yingshan,Zhao Peizhen and Deng Chuolin 2086true Liu Shan,Huang Tao,Xiao Wen and Zhou Yaohe 2085true 3H-colochicine assay (expressed as colchicine binding activity). The results showed that the rate of tubulin synthesis was a peak value period in the brain developing critical period This is exactly a period in which thyroid organization and function are being perfected. Besides, in breeding period, the difference of tubulin content between male and female is significan (P<0.001). The tubulin synthesis is strictly sex dependent (only brain from sex maturity stage). We suggested that the sexual hormone might exert their effect in tubulin synthesis.]]> Guo Shouxiang,Cui Ruzhen,Lu Saojie and Li lange 2084true 60Co irradiation 6.0Gy and/or administration of selenium (1.5mg Se/Kg, Selenite), the activity of glutathion peroxidase (GSHPx), the rate of lipid peroxide (LPO) formation and the incorporation of 3H-TdR in the subcellular fractions of mouse liver and blood were studied. The results showed that the blood and liver mitochondrial GSHPx rose in the 1—4th days, dropped to subnormal level in the 5—12th days and then recovered in the 13—18th days. Se treatment had a markedly protecting effect to mouse on this change.It was also found that the LPO contents of blood and liver subcellular fractions were elevated significantly by 60Co irradiation and small doses of Se had an inhibiting action on such an elevation. These results were supported by the experiment of incorporation rate of 3H-TdR in liver cell. However attention should be paid to a dose-effect relationship between the protecting effect and tbe toxicity of Se. It was found that when the dose of Se was over 0.5mg/Kg, the more Se was given the more LPO was formed.]]> Xiang Rong and Wang Dingnian 2083true 2 was analyzed in order to clarify the mechanism of hemoglobin A2 phenomenon. The paper reports the hemoglobin composition of erythrocyte HbA2.2. The results of two-dimensional electrophoresis showed that erythrocyte HbA2 contains two Hb components: one corresponds to HbA and the other most probably is hemolysate HbA2.3. The results of one-dimensional re-electrophoresis showed definitly that erythrocyte HbA2 is composed of hemolysate HbA2 and HbA.4. Tentative conclusion: HbA2 may be combined with HbA in erythrocyte, i.e. there is probably interaction between the two hemoglobins. This is perhaps the possible cause of hemoglobin A2 phenomenon.]]> Qin Wenbin 2082true 2+/CaM-PK Ⅱ in the presence of Ca2+ and CaM, the maximal phosphate incorporation was 1mol/mol subunit of the 63kD PDE isozyme. 2. The phosphorylated 63kD PDE isozyme was dephosphorylated by calcineurin in the presence of Ni2+ and CaM. 3. The AC50 for Ca2+ of the phosphorylated form of the 63kD PDE isozyme was higher than that of the nonphosphorylated form of the 63kD PDE isozyme.]]> Zhang Guangyi 2081true P. falciparum hybrid peptide antigen gene has been synthesized by the solidphase phosphoramidite method with ABI381A DNA synthesizer. The gene is 216bp in length and divided into 10 fragments to synthesis. All synthetic fragments were annealed and ligated with T4 DNA ligase. The product of synthetic gene was recombinded with phage M13 mp18 vector and transfected to E. coli JM109. The recombinants were screened by spot hybridization with 32)P-label synthetic fragment. The sequence of synthetic gene was analysed by dideoxynucleotide chain termination method. The recults showed that synthesized human P. falciparum hybrid antigen gene was identical with the designed one.]]> Zhong Xionglin,Chen Shirong,Qiu Minyan,Wang Changcai,Min Yongjie and Wang Qisong 2080true 2TPP)in benzene have been investigated by spin trapping-ESR technique. The ESR spectrum with 6 lines was detected in the presence of spin trap PBN, it means that spin adduct is [HTPPPBN]·. The ESR signal with 6 lines was quenched by O2 and NaI, it can be concluded, therefore, that the photolysis process of H2TPP was accomplished via triplet excited state.]]> Zhou Jianwei,Chen Dewen and Xu Guaugzhi 2079true cis-platin. The results shows that after the extrinsic laminin combined with its receptors they can promote the assembly of F-actin, resulting in the increase of the amount of the F-actin, that cis-platin, one of the multiple-targeted antitumor drugs, inhibits the assembly of F-actin after linking with F-actin and that under the action of laminin and Cis-platin, the assembly tends to stability and the amount of the F-actin remains unchanged. This study gives a direct evidence for the opposite action of extrinsic laminin and cis-platin on the assembly of F-actin in ascitic liver cancer cells of mouse.]]> Luo Shirong,Tao Jiaping,He Qihua,Li Hua,Yang Fang and Su Yaxian 2078true 2-·) produced by pyrogallic acid, pH8.4, caused peroxidative damages to cell membranes, such as increase of fluorescent substance on membranes, formation of high molecular polymers (HMP) due to membrane protein cross-linking. However, these were almost not induced after adding a certain amount of Na2SeO3. It is suggested the anti-peroxidation effect of Na2SeO3 on erythrocyte membranes.]]> Qin Dean and He Xuemin 2077true t0.5=75 min (16α-BAP) and t0.5=480min (5α-DTB). When substrate progesterone or 5α-dihydrotestosterone was present in the incubation mixture, the speed of inactivation of the enzyme was slowed down. The labeled enzyme was hydrolyzed by hydrochloric acid, through amino acid analysis, the labeled histidine was identified. The histidine at active site may play important role in the oxidoreductive reaction.]]> Chen Qingxuan and Frederick Sweet 2076true 2+/CaM-dependent PDE activity rather than the PDE-binding activity of CaM in drug-treated cells of both types. However, both MGC-803 and mouse hepatoma cells exposed: to aminophylline (2mmol/L) for 3 days or shorter time showed decrease in PDE-binding activity of CaM. The possible mechanisms concerning TFP effects and their role in regulating cell proliferation of MGC-803 cells were discussed.]]> Zhao Yali,Meng Songniang,Liu Shuren and Lin Zhongxiang 2075true Sun Runguang,Xu Fengxun,Zhang Jing and Tian Suikang 2074true Ab initio calculations on O3, FeO2, Fe(Ⅰ)O2, Fe(Ⅱ)O2, CoO2 and Co(Ⅱ)O2 were carried out by the spin unrestricted Hartree-Fock method. The results show that the Mulliken population of Fe(Ⅱ), central Oc and terminal Or are 24.18, 8.19 and 7.64 respectively for π-system in oxyhemoglobin. This is contrary to the suggestion by Weiss that the Fe-O2 bond is similar to Fe3+O2-. The result appears to fit the experimental data very well and provides good mechanistic evidence for explaining the oxygenation-deoxygenation of oxyhemoglobin in the life of living organisms.]]> Tan Zaiyou and Wu Jian 2073true B/F was developed. 125I-labeled apoE-deficient human HDL3 was prepared as the ligand. 12.5％ of the final PEG concentration could effeciently sediment the 125I-labeled HDL3-membrane complex, and more than 94％ of the total binding was specific (non-specific binding was determined at the presence of 25fold excess HDL3). A saturable and high affinity HDL-binding site on liver membranes with kd 14.5+0.86μg/ml (16.6×10-8mol/L) and Bmax 15.6+6.8μgg/mg membrane protein was found. The Bmax of this HDL-binding appeared to increase as the reactiontemperature rose. The binding activity of this HDL receptor was not inhibited by 0—30 m mol/L EDTA and not activated by 0—5.0 mmol/L Ca2+, as well as insensitive to trypsin. Human HDL3 and rat HDL, but not human VLDL, LDL and albumin, could effectively inhibit the 125I-labeled HDL3 binding to the liver membranes. The 125I-labeled HDL3 binding activity (μg/mg membrane protein) increased with the increasing of the liver membrane purity. These results suggest that there was a saturable and highly specific HDL-receptor on liver plasma membranes, and its biologic properties are different from LDL-and apoE-receptors.]]> Zhang Linhua,Liu Bingwen and Lan Tianhe 2072true Yuan Jingming and Zhai Guotang 2071true Kang Jian,An Tianyi,Chen Xiaolin and Chu Junjie 2070true Liu Yiwei and Liu Bingzheng 2069true 2) in different rates between amino groups of SOD and PEGm2. The purified PEGm2-SOD that remained 80％ residual activity with 30％ modified amino groups was obtained. The fluorescence spectrum and circular dichroism of modified SOD were changed; the results of denaturation by GuHCl and of hydrolysis by pepsin showed that PEGm2-SOD had higher stability than SOD; the results of immunology showed that the immunogenicity and antigenicity of PEGm2-SOD decreased greatly.]]> Wang Jihua,Cao Shugui and Cheng Yuhua 2068true Dong Jianying and Yang Duan 2067true Zhong Ling,Li Wenzhe and Liu Liangshi 2066true bglS gene from B. subtilis. The synthesis of the gene product BglS in E. coli is affected by various hosts, different vectors and orientation of the gene fragment.By protein localization analysis and zymogram analysis of active enzyme molecules, we have found that the enzyme has two active molecules (32 kD and 27 kD) in E. coli and the amount of the enzyme in the periplasmic space is very small. The presence of different active products and the specificity of secretion pattern in E.coli may be due to the difference between E.coli and B.subtilis in the protein processing and translocation.]]> Jiang Hong,Chen Yongqing,Song Daxin and Yang Fan 2065true Li Jun,Zhou Mei and Chen Yuan 2064true E. coli with molecular cloning technique. A 191 bp fragment containing most of the E7 gene was cut out from the HPV 16 genome with the restriction enzyme Dde Ⅰ. The plasmid pATH10 which contains the promoter of the tryptophane operon was chosen as the expression vector. The 191 bp fragment and the vector plasmid were ligated together later and the recombinant DNA formed was used to transform E. coli DH5α. Restriction enzyme mapping and SDS-PAGE protein analysis were done on four transformants. The molecular weight of the newly expressed protein of the transformant was estimated and found to correspond well with that expected theoretically. We concluded that the expressed protein was a fusion protein of TrpE/E732-95aa.]]> Qi Fengju and Xu Qian 2063true in vitro amplification of target sequences via polymerase chain reaction (PCR)and selective oligonucleotide hybridization. 13 samples (26%) contained Ki-ras mutation. 3 carcinomas exhibited 2 distint Ki-ras mutation. The Ki-ras mutation was present in adenocarcinoma, squamous cell carcinoma, adenosquamous carcinoma, well differentiated, moderately differentiated, poorly differentiated human lung cancers.]]> Qin Yang,Sun Zhilin,Zhao Jiansheng,Sun Ning,Bu Hong and Liu Kaifeng 2062true Streptomyces ahygroscopicus were examined by means of different methods, such as ultraviolet spectrum (UV), infrared spectrum (IR), amino acid composition analysis and polyacrylamide gel electrophoresis (PAGE), sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE),0.2mm thin-layer electrophoresis (TLE).The Streptomyces ahygroscopicus glucose isomerase was found to consist of one subunit by SDS-PAGE. The molecular weight of the enzyme determined by polyacrylamide gel electrophoresis in a continuous molecular sieve gradient method was 49700. The pI of the enzyme determined by 0.2mm thin-layer electrophoresis method was 5.2. Large differences were found in the amino acid composition compared with the enzymes from other sources.]]> Li Xiaoping,Huang Youmei,Zhang Feng and Su Pu 2061true IC50 was about 300μmol/L. Lineweaver-Burk plots revealed that IATPD was competitively with ATP and noncompetitively with PGAT to inhibit the kinase activity.]]> Zhang Guangyi,Feng Zhimin,Wei Yong,Zhao Shenghao,Yu Liangli and Wang Lichen 2060true Xu Mingbo and Yao Zhijian 2059true 32P-labelled apo AⅠ, CⅡ and CⅢ recombinant DNA as probes, we determined the mRNA amounts of apo AⅠ, CⅡ and CⅢ in the liver and intestine of rat injected with insulin (1.5U/Kg weight of rat) by dot blot hybridization method. After injecting insulin, the mRNA amounts of apo AⅠ in liver and intestine were obviously decreased, to a lowest level (42%) at 6 h interval and restored to that of control group at 24h interval. Also, after injecting insulin, the mRNA amounts of apo CII in liver and intestine were declined remarkably, to a lowest level (57%) at 12h interval, still lower than that of the control group (79%) at 24h interval. But the mRNA amounts of apo CIII showed no significant changes after rejecting insulha both in liver and intestine when compared with the control group (P＞0.05). The experiment was repeated.12h after injecting insulin, the mRNA amounts of apo Al, CII and CIII in liver and apo A1 and CII in intestine were obviously decreased (45%, 51% and 54%, 35% and 37%, respectively, P＜0.01, n=5) and the mRNA amounts of apo CIII in intestine remained unchange (P＞0.05, n= 5). These resulls revealed that insulin could evidently affect the gene expression of apolipoproteins.]]> Peng Teng,Liu Bingwen and Lan Tianhe 2058true RPL promoter in E.coli following induction at 42℃ for 2h, 4h and 6h, respectively. The proteins in the lysate of bacteria were analysed by using 12.5% SDS-PAGE. The predicted protein 13kD expressed from the cloned gene in E.coli was observed and harvested. Up analysing the antigenic domain of HCMV glycoprotein 52 kD, a polypeptide in length of 25 amino acids was synthesized, of which sequence is Phe-Asp-Leu-Glu-Glu-Ile-Met- Arg-Glu-Phe-Asn-Ser-Tyr-Lys-Gln-Arg-Val-Lys-Tyr-Val-Glu-Asp-Lys-Val-Val. The expressed protein and synthetic polypeptide were used to immunize rabbits respectively, and two kinds of antisera reacted with the expressed antigen and HCMV AD 169 antigen detected by using 125I-labelled protein A. The results of Dot-blot and Western transfer show that the protein expressed from the cloned gene has the same band in size and specificity of antigen as those of the synthetic poly-peptide. By using horse radish peroxidase (HRP) labelled IgG instead of 125I-protein A, preliminary result indicated the diagnostic value of the expressed protein and its antiserum for the HCMV infection.]]> Wu Jun,Chen Junjie,Tang Zeyuan,Gu Jianren,Wan Dafang,Ma Anqing,Qu Shumin and Li Hongnian 2057true 2+, Mn2+, Co2+, Cr3+,Eu3+,Ni2+ may bind to human serum albumin and that the fluorescence of human serum albumin may be quenched by nonradiative energy transfer. In terms of Förster type dipoledipole nonradiative energy transfer the distance between the Trp-214 and the bound Cu2+ in the first copper-binding site was calculated and found to be about 1.20nm.]]> Yang Binsheng and Yang Pin 2056true in vitro in this paper. The results indicated that the tag phase and growth phase were 15 and 120 min respectively, gelatinization rate in growth phase was 0.0060, gelatin reversibility was 13.58%, and collagen fibrils showed typically banding pattern staggered by 64nm, especially for the native fibrils, but the diameters of these fibrils were larger than those in vivo. These data are of importance for studying the formation and regulation mechanism of collagen fibrils in vivo.]]> Fu Zhihou and Zhang Shiyuan 2055true Pseudomona cocovenenans showed that toxoflavin was nonessential activator for the enzyme, and the activation in condition of hypoxanthine as substrate was more marked than xanthine. The activation obeyed the partial activation type. These results provide us a clue for searching toxic mechanism of toxoflavin for human.]]> Ren Weidong,Zhao Naixin,Yun Yunghua,Liu Jinsheng and Liu Dianzhen 2054true Zhang Xinbo,Cui Zhaochun,Tang Naimei and Zhu Zhengmei 2053true Wu Tongle 2052true XU Alian,Zhou Xiaoyin and Donald A. Harn 2051true +-K+- and Ca2+-Mg2+-ATPase were determined in 28 hypercholesterolemia patients and 16 control.The Na+-K+- and Ca2+-Mg2+-ATPase activities were reduced in these patients. Negative correlations were shown between two ATPase activities and plasma TC and LDL-C levels, but positive correlation was shown between Na+-K+-ATPase activity and plasma HDL-C level in all groups. These studies may be important for probing into mechanism of atherosclerosis formation and prevention and cure of this disease. This problem is discussed in the paper.]]> Yang Shaomin,Chen Yuhua and Zhang Xiaoxian 2050true Yu Xiaobo,Wang Ling,Jing Xiaotang and Wang Zhong 2049true Sun Runguang,Wang Dehua,Wang Mingzheng,Wan Xiuqing and Zhang Jing 2048true 2-cytochrome c reductase.No any inhibitory effect of this two compounds on the terminal oxidase had been observed.The hydroxy groups in the rings of both compounds may be involved in the inhibitory effect through their disturbance on the hydration and dehydration of ubiquinone.]]> Guo Dahai,Xu Jianxing and Gu Lianquan 2047true E.coli was coupled chemically with activated polyethylene glycol(PEG).The modified rhCu/Zn SOD,with apparent molecular weight about 60000 daltons,has longer half-life(15 hours), and the remaining enzyme activity is more than 80 per cent. Its stabilities against acid, alkali and heat were increased significantly. The relation between the amount of PEG used and the remained enzyme activity was also reported.]]> Luo Xunyi,Wang Jingyi,Xie Bangtie,Li Zanqing,Liu Xiaoling and Chen Xiaosui 2046true Fusarium tricinctum,which was similar in structure with that from Fusarium species found in grains from Kaschin-Beck disease area.When the concentration of the T-2 toxin rose to 0.01 ppm,the collagen microfibrils in extracellular matrix of chicken embryo chondrocytes were significantly decreased,and the activities of cytochrome c oxidase and H+-ATPase of mitochondria were obviously lower than that of the control. The results showed that 0.01 ppm the T-2 toxin induced the changes of structure and function of chicken embryo chondrocytes. The methods reported provided an effective measure to examine if the factor which induced the Kaschin-Beck disease attacked on the target tissue——chondrocytes directly.]]> Li Shengguang,Sun Shan,Wu Lianying,Ji Huifen,Hong Jie and Lin Zhihuan 2045true 3H-TdR into DNA of 3T3 cell. The apparent molecular masses of aFGF is 13.2 kD and bFGF 15.2—15.8kD.The aFGF has an isoelectric point range of 5.0—6.0 and the amino acid composition consistent with that of standard aFGF.]]> Xue Yanning,Wang Huixin,Zhou Tingchong,Shao Ningsheng and Wang Fang 2044true Carassius auratus induced by exposure to cadmium chloride and characterized.It shows very similar to mammalian metallothioneins in all of its properties.The MT of the fish contains high cysteine contents(30%)and with an absence of aromatic acid residues.The thiol groups of the cysteine residues combine with the cadmium in SH/Cd molar ratio of 3∶1 and produce a characteristic absorption maxium at 250 nm,minimum at 280 nm.The metal analyses reveal only traces of zine and copper in addition to cadmium. The molecular weight is determined to be about 10 000 by gel filtration on HPLC and SDS-PAGE method. The isoelectric point of fish MT is demonstrated to be about 5.6.]]> Ren Hongwei,Wang Wenqing,Ru Binggen,Li Lingyuan,Pan Aihua and Wang Yingyan 2043true Ashbya gossypii and two kinds of its mutant strains CMG and JMG,which were obtained by γ-ray and Helium-Neon laser irradiation,respectively.The experiments resulted in the conclusions that AG contains five chromosome DNAs;and that no double-stranded breaks or loss of large DNA fragments but alkaline labile sites existed in both the mutant strains.]]> Yang Suhong and Ge Zhongliang 2042true Sun Jishan,Zhang Li and Yue Jiachang 2041true Mycobacterium tuberculosis is amplified by polymerase chain reaction and we get a specific 158 bp DNA fragment of Mycobacterium tuberculosis.The DNA of Mycobacterium bovis is also amplified,but other thirteen mycobacteria are not.The DNA fragment is recovered and used as a DNA probe,It can hybridizes with DNA of Mycobacterium tuberculosis and Mycobacterium bovis,but not with the DNA of Staphylococcus aureus,Bacillus pyocyaneus and other mycobacteria.This result suggests that the Mycobacterium tuberculosis and Mycobacterium bovis can be detected by PCR and the amplified 158bp DNA fragment can be utilized as a molecular probe. This probe can examine the Mycobacterium tuberculosis and Mycobacterium bovis and distinguish them from other mycobacteria.]]> Ding Xiaohua,Yang Ping,Liu Weiqian,Tan Yun,Zhao Wenxian,Cai Jianbin,Mei Guohua and Pan Huike 2040true 125Ⅰ-labeled apoCⅢ to human hepatic plasma membranes revealed saturation binding to membranes with a Kd of 0.31μmol/L(3.1×10-7mol/L) and binding maximum of 1.74μg/mg of membrane protein.In displacement studies using unlabeled apoCⅢ and isolated lipoproteins HDL,LDL and VLDL,only apoCⅢ and VLDL effectively competed with 125Ⅰ-apoCⅢ for membrane binding sites. The binding of 125Ⅰ-apoC l to human liver plasma membranes was Ca2+-independent and was abolished when plasma membranes were treated with trypsin. The characteristics of apoC Ⅲ-binding sites of mouse liver plasma membranes was similar to that of human liver plasma membranes with an exception of binding maximum of 1.52μg/mg of membrane protein.]]> Fang Dingzhi and Liu Bingwen 2039true Wu Yuzhang and Zhu Xihua 2038true Escherichia coli with a recombinant plasmid prepared previously in our laboratory.The E7 fusion protein produced and purified by gel electrophoresis was used as antigen to immunize rabbits for preparing anti-E7 protein antisera.The antisera were used to stain the tissue sections often cases of uterine cervical cancer with an immunohistochemical technique(colloidal gold labeled stain).The E7 antigen was seen in the cells of 6 cases of cervical carcinoma as black granules under light microscope. The E7 protein was found in the nucleus of positive cells, mainly on the nuclear membrane, arranged in a circle. Only a few silver stained granules were seen in the cytoplasm. We believe that the intracellular distribution of the E7 protein indicates that there is high expression of the E7 gene in the HPV infected cells of the cervical cancer; and the fact that E7 antigen is a nuclear protein suggests that E7 gene is an oncogene of HPV 16.]]> Xu Qian and Qi Fengju 2037true Phormidium Iuridum.Monomerization mostly results in the disappearance of characteristic absorption peak 650nm,and in the changes in relative intensities and positions of Raman bands of phycocyanobilin chromophores.As a result,in the monomeric allophycocyanin,the chromophores adopt cyclohelical conformations,which close to these of free chromophores in vitro; and in the trimeric allophycocyanin, fully extended conformations of the chromophores can be marked by the main Raman peak around 1645 cm-1, the behaviour of which is in Rood agreement with the Avis/Auv, absorption ratio.]]> Xia Andong,Zhu Jinchang,Jiang Lijin,Zhang Xinyi and Kong Xianggui 2036true Lei Dianliang,N.Sigimoto and M.Matsuda 2035true Km of 9.8×10-5mol/L with butyryl-thiocholine iodide was determined. SDS and polyacrylamide gel electrophoresis showed that the choline esterase existed in the different polymers composed of the same subunit. The molecular weight of subunit was 78000, and the subunit had the activity of the whole choline esterase.]]> Guo Shengqing,Cao Shugui and Cheng Yuhua 2034true 2+-induced oxidative modification of LDL.As evidencedby the lipid peroxide content,lipofusion and the elec trophoretic mobility,vitamine C could inhibit the oxidative modification of LDL for 24h. The extent of the inhibition depended on vitamine C concentration. These findings indicate that vitamine C is a potentantio-xidant in vivo, it can protect LDL from oxidative modification. Hence, vitamine C can prevent and treat atherosclerosis.]]> Wang Dongxiao,Zhou Mei and Chen Yuan 2033true 20 exibites stronger inhibition on CaM-activatedphosphodiesterase than TFP.IC50=5μmol/L.Flourescence analysis indicats that the asso-ciation constant (Ka) for the interaction between CaM and compound D20 is 2.64(μmol/L)-1 and compound D20 molecule possesses 2 binding sites for CaM.Binding style is discussed.]]> Zhang Yuan,Hu Zhuoyi,Zhao Ye and Cai Huimin 2032true I of 4.2 and an apparent Mr ofabout 92kD,as judged by polyacrylamide gel electrophoresis,which consists of two non-identical glucopeptides with Mr of 64 kD and 27 kD, respectively. The purified GGTPs were used to produce antibodies in rabbits. The antisera with titers of 1:16 were observed to be capable of being absorbed by the constituents of the novel GGTP, Ⅱ and Ⅱ', in sera of patients with hepatoma. Furthermore, they could form in the double diffusion test a smoothly fused precipitating line with two kinds of purified antigens. These results show that GGTPs from human hepatoma and fetal liver tissues are of immunological identity.]]> Guan Saifang,Xu Kaili and Chen Fuhua 2031true Xu Di,Jiang Chunlei,Zheng Zhongcheng,Fan Peifang,Sun Lanying,Liu Xinyuan,Song Chaoyou,You Zhendong,Wang Chenghai and Lu Changlin 2030true Cai Guiying and Zeng Wenjie 2029true Li Yuanxun and Ye Qingling 2028true Lu Bin,Yie Ning and Wei Yu 2027true 2＋ in every FP molecule and Ca2＋ ions are located near the Trp.When Tb (Ⅲ)-FP was excited by 222nm and 233nm, it is possible that energy transfer occurs are as follows: Trp→Tb3＋,Trp→Tb3＋→Phe,Tb3＋→Phe,Tb3＋→Trp.These results indicate that Phe may combine with Tb (Ⅲ).The results of calculations indicate that the distance between Trp and Tb (Ⅲ) is atout 5.56Å.]]> Xia Wensheng,Lu Jingfen,Liu Hongyu and Liu Qingliang 2026true Wu Meiying and Mcintyre D 2025true ＋/H＋ exchange upon the measurements of proton transport are studied.]]> Zhang Guojiang,Wang Jia,Yao Junlan and Liu Xiaoqi 2024true Xiu Bo,Yu Xin,Li Huifang and Zhou Airu 2023true Sun Congmei,Wang Dong and Zhou Airu 2022true E.coli cytoplasm. The IBs were isolated by centrifugation and sonication. CH925 were refolded and reoxidateo in a glutathione redox system following denaturation of the IBs in 7mol/L guanidine hydrochloride. The specificactivities of IL2 and IL6 assayed by CTLL-2 and 7TDl cell line were 2.2×106U/mg and 2.3×108U/mg, respectively,following renaturation. The renatured CH925 was chromatographed through DEAE-Sepharose 6B column and Sephacryl and column. The active fractions were pooled and applied to HPLC with reversed-phase C-18 column, while CH925 was eluted through 10%-70% acetonitrile gradient. It showed only one pratein peak and SDS-PAGE confirmed that the purified CH925 was almost homogeneous.]]> Zhao Chunhua,Ling Shigan,Mao Ning and Tang Peihsien 2021true Sun Naien,Wang Yan,Xie Wei and Zhu Dexu 2020true e. implying that MT can interact strongly with membrane. The experiments carried out in vitro demonstrated that MT could be absorbed on the surface of erythrocytes. In addition, rab-hits were in Jected s. c. with CdCl2 to induce the biosynthesis of MT, the presence of MT mainly in the erythrocyte is first suggested after chromatographic separation of plasma and blood haemolysates from Cd-loaded rabbits. It is claimed that a dynamic equilibrium could be established between MT absorbed on the surface of erythrocytes and presence in plasma. The significance of the above findings is disscussed in brief.]]> Zhang Baolin,Lu Jingfen,Wang Wenqing,Ru Binggen and Tang Wenxia 2019true ＋-ATPase has been successfully expressed in E.coli. The fragment of 58kD subunit gene was obtained by polymerase chain reaction (PCR). A clone encoding 58kD subunit was obtained by directly joining PCR product into the plasmid for expression by T7 RNA polymerase (PET). Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blot analysis of cultured transformantsdemonstrated high expression of 58kD subunit gene. The product of 58kD subunit accounted for 50% of cytoplasmic proteins.]]> Zhang Ying,Peng Shengbin,Stone D. K. and Xie Xiaosong 2018true E.coli ompA signal peptide and mature sGH was constructed and confirmed by restriction enzyme analysis and Southern blot hybridization with the oligonucleotide probe. Upon induction of IPTG, the sGH was expressed in E.coli cells and was secreted into the E.coli periplasma.]]> Zhang Tonghai,Song Shiduo,Zhao Weicheng,Chen Kunming,Qi Wei,Hu Wenzhi,Wang Peifu and Fang Peihua 2017true E. Coli. The fragment of the gene of 70kD subunit was obtained by polymerase chain reaction (PCR) from an isolated cDNA encoding the 70kD subunit of the bovine brain vacuolar proton pump. The fragment of the gene of 33kD subunit was also obtained by PCR from the bovine brain cDNA library. Two clones encoding 70kD and 33kD subunits were individually obtained by directly joining the PCR products into PET vector. Sodium dodecyl sulfate (SDS)-poly-acrylamide gel electrophoresis (PAGE)and Western blot analysis of cultured transformants demonstrated high expression.]]> Zhang Ying,Peng Shengbin,Stone D. K. and Xie Xiaosong 2016true 2+, Mg2+-ATPase). These aterations perhaps related to the increase of lipids hyperoxidation induced by free radical and exchange of lipids.]]> Dong Chuanren,Yu Xuegang and Wang Xuejun 2015true Chen Suzhen and Han Jisheng 2014true E. Coli with the novel TNF expression plasmid produced high yield of novel TNF, whose cytotoxic activity to L929 cell was 103 higher than recombinant human TNF.]]> He Xiaolong,Chang Jinli,Cai Wucheng,Yu Hong,Lu Qun,Zhao Shouyuan,Wang Chenghai,Lin Baocheng and Zhu Henian 2013true Chen Changhu,Yao Kaitai and C.C.LING 2012true E.coli was covalently linked with an amphipathic molecule poly-(styrene-co-maleic acid) butyl ester (SMA) via amide linkage. When 42% free amino groups of the enzyme were modified. 88% remained enzyme activity was obtained.The results of circular dichroism of rhCu/Zn SOD and SMA-rhCu/Zn SOD indicated that the structure of the modified rhCu/Zn SOD was scarcely changed. Its biological half-life in blood was prolonged 22 times.and its abilities to resist pepsin and trypsin were increased significantly.]]> Luo Xunyi,Wang Jingyi,Xie Bangtie,Li Zhanqing,Liu Xiaolin and Chen Xiaosui 2011true Bacillus sp.O74.The enzyme was purified 27.9 fold by sephadex G-100 gel filtration,ion-exchange chromatography and hydrophobic interaction chromatography.The enzyme was characterized by demonstration of optimum activity at 50℃ and pH 7.0.and its molecular weight of 52 500 determined by gel filtration.The pH range of the enzyme showing the activity is from pH 4 to 12,and at pH 9 and 10,it can keep 80% and 70% of the maximum.activity respectively.The enzyme was stable in the presence of the most metal ions, surface active agents and auxiliaries.]]> Wang Dong,Song Guijing and Gao Peiji 2010true Zhang Jianzhong,Zhu Yuanxiao,Wang Jianan and Zheng Jianqiang 2009true 1 and P2. The recovered PCR product was flushed by klenow fragment and inserted into pGEM-3Zf (+) vector pretreated with SmaⅠand calf intestinal alkaline phophatase. The recombinant plasmid was used to trans-form competent cell JM 109. The positive colonies were directly screened on indicator plates. The recombinant plasmid DNA and insert fragment isolated from four positive colonies were analyzed by digestion with EcoR Ⅰ，Kpn Ⅰ and Taq Ⅰ.The different coding sequences including MBP exon Ⅰ—Ⅶ, Ⅰ—Ⅲ，Ⅲ—Ⅶ and Ⅰ—Ⅴ were amplified from these clones with their corresponding nested sets of primers respectively. These results show that these cDNA clones contain full-length coding sequence for 21.5kD human MBP.]]> Chen Junjie,Wang Ruohan,Cheng Hanhua,Chen Pu,Li Changlong and Yang Luchuan 2008true cis-trans isomerase (PPI) in vivo is a limited procedure in protein folding. In order to study the catalyzing activity of PPI on the refolding of recombinant proteins in vitro,PPI is purified from pig kidney.and is investigated the effects of the enzyme on catalyzing the refolding process. Results indicate that PPI increases the folding rate without increasing the correct folding ratio and specific activity.and PPI has a high catalyzing activity even at very low concentration.]]> Xu Mingbo,Meng Wenhua and Ma Xiankai 2007true Zhu Ping,D.MANN and R.GREENBERG 2006true Zhang Jingxia,Huang Ping,Xu Shiwen,An Lizhi,Yao Huiying,Xiao Yanan,Pan Juxiang,Chao Zhiyu,Zhu Jieqing and Wu Xiankang 2005true Hu Kunsheng,Shi Hua,Huang Ying and Yu Bi 2004true Yan Jiaqi and Xie Wenzheng 2003true 1F0-ATPase activity assay. 31P-NMR spectra measurement. fluorescent probe NBDPE detection. packing and fluidity of membrane lipids and intrinsic fluorescence measurements can be summarized as follows: ADM induces the phase transition of mitochondrial membrane lipids at first. as a consequence affecting on the lipid packing and fluidity of the lipid molecules and then influencing the conformation of the F1F0-ATPase and finally resuiting in the decreasing of the enzymatic activity. And Mg2+ can protect all the effects induced by ADM thus reducing the harmful effect of ADM.]]> Lin Zhihuan,Li Shengguang,Cao Maosun,Chen Yunjun,Feng Chaoyang and Deng Junpeng 2002true 2+ and Mg2+ concentration(c (Ca2+)i and c(Mg2+)i) of alveolar macrophages in rabbit lung after incubation with sodium selenite were determined by SPEX AR-CM-MIC system.The results showed that high concentration sodium selenite(≥10-4mol/L) had the cytotoxicity.The poisonous effect of selenite on macrophage led to the increase of c(Ca2+)i and c(Mg2+)i.High concentration selenite had some suppressive effect on the Ca2+-ATPase activity of erythrocyte membrane.]]> Wen Herui,Wang Jinxi,Zhu Weihua,Li Ruiyan and Chen Rongsan 2001true Wang Zhaohui,Shan Weisong and Wu Xiru 2000true 32P] ATP bv T4 polynucleotide kinase.When this modified thymidine located in the middle of oligonucleotide,50% activity of T4 polynucleotide kinase was inhibited.When this modified thymidine located at the 5'-terminus,only 2% activity remained. Comparatively,those modifications did not affect the activity of T4 RNA ligase.]]> Zhang Xiaolan,Lu Changde and Qi Guorong 1999true Lu Bin,Liu Cuihua,Shang Heyong and Xu Jianxing 1998true Ziziphi spinosae (ZS) could markedly and dose-dependently decrease MDA content，elevate SOD. activity and increase membrane fluidity. It proved the effects of ZS against lipid peroxidation induced by anoxia-reoxygenation.]]> Wan Huayin,Ding Li,Kong Xiangping and Chen Xingjian 1997true RPL promoter of the expression plasmid pBV220 by using of DNA recombinant technique.This recombinant plasmid can express hIFN-γ stably with high efficiency(up to 44.4% of the total bacterial soluble protein) in E. coli DH5α through thermal induction.The bioactivity assay showed that 0.45×107～2.34×107U rhIFN-γ were expressed in every liter of bacteria culture after preliminary purification and refolding procedures.]]> Qu Chengkui,Wei Handong,Yu Yongtao,He Fuchu and Wu Zuze 1996true P<0.01),and the falling of relative percentages of GM1 and GD1b of which were magnificent (GM1:P<0.05,GD1b:P<0.01);②Among monoamine neurotransmitters，the level of NE and DOPA descended and that of DA and HIAA increased (P<0.01).The results point out:after repetitive hypoxia，the cell membrane in centrical nerve tissues is injured in some degree and thus may affect the whole process of synthesizing，releasing，reabsorbing and storing of monoamines. There is association relation between changes of gangliosides and monoamines when hypoxia occurs.]]> Yang Dianer,Pan Ying,Li Aihua,Huang Rubin and Xiong Ying 1995true Chu Jiayou,Wu Min,Shen Ziwei and Zhao Nanming 1994true Yan Guangtao,Hao Xiuhua and Li Zhenjia 1993true Wang Zhiyong,Zheng Zhongcheng,Sun Lanying and Liu Xinyuan 1992true Jin Guichang,Zhang Jingzhi,Zhou Guirong and Zheng Zhuying 1991true cis-elements were compared with those in pSV2-dhfr.]]> Long Jianyin,Zhang Hongquan and Wang Huixin 1990true Hu Meiru,Ji Yanshan,Shu Cuiling,Chen Lijun and Shen Beifen 1989true c were significantly higher in hypoxic group than that in control(P<0.01).The increase in the ratio of h0/h-1 was well correlated with the enhance in hypoxia group. It was also found that MT could inhibit the lipidation in cerebral cortex cells.]]> Zhang Qian,Wu Benjie,Yu Guifen and Lu Jingfen 1988true Mr 5000),activated by cyanuric chloride and nitrophenol carbonate respectively,was used to modify subtilisin Carlsberg. The enzyme after PEG-modification maintained its catalytic activity both in aqueous solution and in 0rganic solvents,and an enhancement in activity and stability in aqueous solution was achieved:the kcat increased accompanied with a lower activation energy,while Km was not changed;the stability of the enzyme against both temperature and pH was greatly enhanced，but the optimal reaction temperature was unchanged. The pH dependence of the modified and unmodified subtilisin has also been compared.]]> Yang Zhen 1987true 2+-ATPase from rabbit skeletal muscle sarcoplasmic reticulum vesicles was solubilized by nonionic detergent C12E8 and purified in a reactive red-120 agrose affinity column.Determined by 7.5% SDS-PAGE,the purity of Ca2+-ATPase was increased from 65% to 99% and the purified Ca2+-ATPase exhibited higher ATP hydrolysis activity.]]> Tu Yaping and Xu Hong 1986true Viscum coloratum lectin (VCL) was first separated from extracts of the stems and leaves of Viscum coloratum Chinese,an ordinary traditional medicine beijisheng,by affinity chromatography on acid-treated Sepharose 6B and gel filtration on Sephacryl S-200,with Mr=52 000,pI=10.5.The LD50 was 4.4μg/kg for mice by intravenous injection,It's a toxic protein obtained from the plant with higher toxicity to mice. The crude extracts of the poant also have strong toxicity,because it's LD50 was 200 mg/kg for mice by intraperitoneal injection and to be expressed as the amount of dry materials.]]> Li Gee,Liu Qing and Yuan Xiaomei 1985true Wan Fusheng and Nan Guohua 1984true 3H]-TdR incorporation and the determination of mortality of cells,the effect of CAIF on the DNA synthesis and cytotoxicity of all cell lines were investigated and the inhibitory effect of CAIF on angiogenesis was measured by the experiment of chorioallantoic membrane of chicken embryos. The results showed that CAIF inhibited the endothelial cell strongly and Hela cell slightly;but not QGY7703 cell，human skin fibroblast，and mice myeloid cell. It also inhibited the angiogenesis in the chorioallantoic membrane of chicken embryos significantly. All these results suggest that CAIF has the inhibitory effect on angiogenesis. It is a specific angiogenesis inhibiting factor，and may be a useful drug on tumor therapy in some day.]]> Shen Xianrong,Jia Fuxing,Yu Zhijie,Xu Hui,Wang Ling and Chen Qi 1983true Zhou He and Yang Qijian 1982true Chen Youiun,Zhu Feng,Ma Helin and Kang Yufan 1981true 3 on the metabolism of phosphoinositides of human monocytoid leukemia J6-2 cells has been studied using the incorporation of[ 32P]Pi and [ 3H]inositol into them. The results show that GM3 inhibited the incorporation of[ 32P] Pi and [ 3H] inositol into phosphatidylinositol (PI),stimulated the incoporation of those into phosphatidylinosito1-4,5-bisphosphate (PIP2) and [ 32P] Pi into phosphatidylcholine (PC), inhibited the incorporation of [ 3H] inositol into inositol phosphates (IP3,IP2, IP )，and of [ 32P] Pi into phosphatidic acid (PA) in a dose-dependent manner. These results suggest that GM3 inhibits the PI turnover of J6-2 cells.]]> Ma Keli,Liu Yan and Cui Zhaochun 1980true Huang Rubin,Yang Dianer,Li Aihua,Pan Ying,Jin Youyu and He Jianyu 1979true Lu Baisong and Huang Peitang 1978true o-diphenols to o-quinones in organic solvents has been investigated with addition of different amounts of water and at various temperatures. The results have shown that tyrosinase possesses high activity in organic media,the enzyme activity is mainly controlled by the water activity in the reaction system,and it can increase with the increase of reaction temperature as long as the amount of water bound to the enzyme molecule is properly controlled. Salt hydrates are effective for such control in nearly non-aqueous conditions.]]> Yang Zhen and D. A. ROBB 1977true 4 and HI-6. in reactivating sarin-,soman-,tabun- and VX- phosphorylated acetylcholinesterase (AChE) imply that there are factors which hinder phosphorylated enzymes from being reactivated by some oximes before aging. To investigate this phoblem,a comparison of in vitro reactivation of these phosphorylated enzymes by H series oximes,HI-6,HGG-42 and their 4-oxime isomers.was made. Results showed that,in tabun experiments,although there was no reactivation by the two 2-oximes (HI-6 and HGG-42)，28%-45% reactivations were observed by their 4-oxime isomers. In soman case the results were opposite. HI-6 and HGG-42 showed remarkable reativation. but their 4-oxime isomers were ineffective. For sarin- and VX- phosphorylated AChE, all oximes showed high effctiveness. These results indicated that there are conformational differences among the active centre of the above phosphorylated enzymes. The influences of AChE allosteric agents showed that C10 significantly increased the effects of TMB4 in sarin, soman and VX experiments, however. it reduced the effect of TMB4 significantly in the tabun case. Propidium has no influence on the reactivation of sarin, soman- and VX-phosphorylated AChE, but it significantly reduced the effect of TMB4 on reactivating tabun-phosphorylated enzyme. These results confirmed that the conformation in the active centre of tabun-phosphorylated enzyme is obviously different from that of sarin-，soman- and VX- phosphorylated enzymes.]]> Luo Chunyuan,Li Zhixiu,Xia Shuquan,Sun Manji and Yang Jinsheng 1976true 21 expression in non-cancer lesion and carcinomas of the stomach was quantitativaly studied by flow cytometry,and relation of ras P21 expression to DNA ploid and cellular proliferation index were inquired.The results showed that ras P21 expression was higher in carcinomas than in non-cancer lesion one’s of thee stomach.In non-cancer lesions,ras P21 expression in precancer lesion was higher than in chronic atrophic gastritis,and the raiser of fluorescence index in ras P21 expression was found with raiser of histological grade. DNA ploid was associated with ras P21 expression.]]> Zuo Lianfu,Hu Junlan,Liu Jianghui,Guo Jianwen,Gao Guodong and Qi Fengying 1975true Xiao Zhiqiang,Yao Kaitai,Liu Wei and Lu Guangxiu 1974true 3+-containing transport buffer was monitored fluorometrically. The technique is highly sensitive, reproducible and much convenient as well.]]> Feng Chang,Guan Zhiying and Tu Yaping 1973true E.coli with high level.The expression product was purified by Sephadex G-50 gel filtration and converted to the mutant of human insulin by trypsin and carboxypeptidase B.The insulin mutant,purified by DEAE-Sephadex A-25,has an expected amino acid composition.The receptor binding activity and biological activity of this mutant are the same with that of standard porcine insulin.]]> Chen Laitong,Tang Jianguo and Hu Meihao 1972true 17 against carcinoembryonic antigen (CEA) by using a synthesized chelate-N1-(p-isothiocyanatobenzyl)-diethylenetriamine-N1,N2,N3, N3-tetraacetic acid Eu is described.The Eu-labeled C17 was separated from excess reagent by gel filtration on a column of Sephadex G-50.The fluorescence and ultraviolet absorption for the fractions were monitored and the curves of fluorescence and the optical density were all the same.The specific activity for the two tubes of the labeled-C17 at the protein peak was 5.2 and 9.8 Eu3+/C17 molecule respectively and the recovery of the labeled-C17 was 64%. The labeled-C17 was stable for 6 months at least. The standard curves of time-resolved immunofluorometric assay using the labled-C17 showed excellent linearity and slope and had been used in the assay of CEA in serum.]]> Zhao Qiren,Zhang Fuhua,Liu Jie,Lin Han,Li Meijia,Zhuang Xianglian,Chen Ai and Hu Bi 1971true Li Peifeng and Fang Yunzhong 1970true Vm)and Michaelis constant(Km)increased nearly 6 and 3 times,respectively in microemulsion compared with that in aqueous solution.]]> Li Ganzuo,Ren Xuezhen,Liu yan,Wang Funing and Zheng Mongying 1969true Schizophyllum commune. Gas chromatography,periodate oxidation,methylation analysis and other experiments showed that the main chains are composed of (1-3 )linked glucose residues one third of which have side chains in position 6.The result of oxidation with chromium trioxide and IR suggested that all residues of Scl are βconfigurations. The conformation of Scl was studied by complex-formation with Congo Red,and by the measurements of optical rotatory and viscosity. Scl mainly a single-helical conformation in water solution at lower temperature. If the side chains are removed by periodate oxidation，the proportion of multiple-helical conformation will increase. The helical conformation changes into disorder coli in acid，base，and at slightly higher temperature.]]> Zhou Yifa,Zhang Liping,Yang Qingyao and Zhang Yishen 1968true 3 on the endogenous protein phosphorylation of human monocytoid leukemia cell line J6-2 cells was studied.Addition of 100 μmol/L GM3 to the particulate fraction of J6-2 cells enhanced the endogenous phosphorylation of several proteins whose relative molecular mass were 180,87,78,67,43 and 31kD while addition of 100 μmol/L GM3 to the cytosol fraction of J6-2 cells stimulated the phosphorylation of several proteins whose relative molecular mass were 87 and 56kD,but could inhibit the phosphorylation of 70 and 43kD proteins. Because GM3 has been reported to be able to induce J6-2 cells to differentiate along the monocyte/macrophage route by long term incubation with GM3 (up to six days)，so the authors postulated that protein phosphorylation induced by short term GM3 treatment might be the early signal of differentiation of J6-2 cells.]]> Dong Zheng,Ma Keli,Han Rui and Cui Zhaochun 1967true Tang Rong,Ma Congcong,Mao Yumin,Xu Yaou and Gong Rongci 1966true in vitro amplification by polymerase chain reaction of a relatively conserved portion of L1 ORF of HPV.A specific fragment of 394-552 bP and 139-15O bp was amplified by PCR from clinical specimens(cervical carcinoma, genital condylomata,ovarian adenocarcinoma etc.)containing DNA from various HPV types. The digestion pattern of restriction endonuclease Rsa Ⅰ analysis of PCR products by PAGE can be used for typing. This method is simple，sensitive，specific and is suitable for detection and typing for genital HPV types in clinical samples.]]> Chen Song and Lu Yijuan 1965true -4 to 1.3×10-2mol/L.]]> Liu Haiying and Deng Jiaqi 1964true Chen Yuhua,Song Jindan and Li Dawei 1963true Mr of 41.3 ku and pI of 4.9. The optimum temperature is 37～40℃. The optimum pH is 7.0. Using adenosine or 2-deoxyadenosine as substrate the apparent Km of the enzyme is 83 μmol/ L and 61 μmol/ L respectively. The enzyme activity can be inhibited by p-chloromercuric benzoic acid while partially restored by dithiothreitol. ADA activity was decreased by anti-calf ADA IgG and anti-thymus ADA IgG.]]> Luo Shiwen,Li Jinsheng,Shen Jingxian and Zou Guolin 1962true 3 and 106 after modification respectively.]]> Li Weizhong and Miao Fangming 1961true Feng Dan,Yuan Yong,Zhang Yingmei,FengLan,Fan Hui,Di Chunhui,Song Quansheng and Ma Dalong 1960true 3H-TdR incorporation into DNA of SMC (P>0.05),(2) LDL and VLDL showed the obvious stimulating effects (P<0.05);(3) OX-LDL, OX-VLDL and OX-HDL increased siginificantly 3H-TdR incorporation into SMC DNA, respectively (P<0.01). These results suggest that the atherogenic roles of LDL, VLDL, OX-LDL, OX-VLDL, OX-HDL are closely related to their stimulating effects on DNA synthesis and the proliferation of the arterial SMCs.]]> Wang Haochuan,Liu Bingwen and Fu Mingde 1959true He Jinan,Jiang Lijin,Zhao Jinquan,Bi Zhichu and Jiang Long 1958true Zheng Xiguang,Wang Hezhou,Yu Zhenxin,Gao Zhaolan,Zhu Jinchang and Jiang Lijin 1957true sis-inducible element)，but seems to have no effect on any DNA-binding factor that recognizes GAS. Thus, IL-2, IL-4 and γ-IFN induce their signals to nucleus by different path-ways in CTLL-2. The data also show that treatment of Hut-102 cell with IL-2 or IL-4 can weakly activate a DNA-binding factor specifically to recognize GAS.]]> Zhu Jinfang,Zheng Zhongcheng,Sun Lanying,Guo Liying,J.THèZE and Liu Xinyuan 1956true 2+ signaling in single intact living cells are described using laser scanning con focal microscopy (LSCM) and the highly fluorescent Ca2+ -sensitive dye, fluo-3/AM. The duration, dye concentration, and requirement for Pluronic are determined empirically for the loading conditions. It is especially important for the proper intracellular dye concentration to provide adequate signal strength for detection while not to disturb normal intracellular physiology. For example, in C57BL/6J macrophages, it was found that cells cultured on glass cover slips were incubated for 1 h at 37℃ with 6 μmol/L fluo-3/AM, resulting in excellent LSCM imaging of intracellular Ca2+.The similar results have been successfully obtained by this method for other types of cells such as vascular smooth muscle cells, Chinese hamstar oocytes. It may provide an available, visual experimental approach for investigation into the dynamic changes of Ca2+ signal and their relationship to transmembrane Ca2+ gradient in living cells under the physiological and pathological conditions.]]> Yang Xiaoyi,Yan Kun and Huang Youguo 1955true 2+ channels were recorded on cultured ventricular myocardiocytes of neonatal Wistar rats by cell-attached patch clamp technique. T-2 toxin at dose of 10 mg/L significantly inhibited the activities of the three types of Ca2+ channels. The open time of Ca2+ channels was shortened and the openstate probability was decreased, whereas there were no significant variations in the amplitude of ionic current flowing through the Ca2+ channels. Compared with verapamil and Bay K 8644, the effects of T-2 toxin on the Ca2+ channels were similar to the blockade effect of verapamil. It was suggested that the cytotoxic action of T-2 toxin may be due to the damages of cell membrane .]]> Peng Shuangqing,Zhang Wenjie and Yang Jinsheng 1954true t-test. From the point of view of cloveleaf structure, the method of randorrT stacking is the best, the second are the optimal stacking and Zuker's algo-rithms and the worst is the maximum base pairing. Finally, the difference between two methods of minimum free energy was analysed.]]> Li Wuju and Wu Jiajin 1953true Chen Hong,Hua Ling,Wang Yue,Zhao Kun,Yan Maogong,Dong Yicheng,Yu Mingkun and Yao Qizhi 1952true 2-cytochrome c reductase has been prepared by a column of calcium phosphate.The activity of this enzyme is lower and the cytochromes contained in this enzyme is partially (about 53 % of cytochrome b and 83 % of cytochrome c1) reduced. The reconstitution of the lipid-depleted QH2-cytochrome c reductase with lipids can not only recover the activity but also induce the reduced cytochromes retain to its oxidized state. This result indicates that a special interaction between the lipid and protein is important for keeping the enzyme in its native conformation.]]> Zhang Yixin,Shang Heyong and Xu Jianxing 1951true Comamonas testosteroni was purified to homogeneity upon SDS-PAGE by using sonic crushing, ammonium sulfate fractionation, molecular sieve chromatography, calcium phosphate gel chromatography and ion exchange chromatography. MOB4-HOase has been purified about 21 fold in specific activity with about 30% yield. This enzyme is a FAD-monooxygenase to catalyze m-hydroxybenzoate to protocatechuic acid.]]> Chen Rui and Keiichi Hosokawa 1950true Xiao Zhijian,Yang Xizhen,Jiang Mengjun and Huang Xuquan 1949true Astragalus (TFA), one of the natural anti-oxide agent isolated from Astragalus monghlious, on DNA strand breaks induced by the hydrogen peroxide and gamma rays has been investigated with the fluorometry assay of DNA unwinding (FADU). The results showed that the TFA has different degree of protective effect on DNA strand breaks caused by hydrogen peroxide and gamma rays respectively. It also showed some protective effect on DNA strand breaks induced by these two damage factors,when the concentration of TFA were 0.4 g/L and 0.6 g/L, respectively. When its concentra-lions were increased to 0.8 g/ L and 1.2 g/L, TFA could effectively protect DNA from hydrogen peroxide and gamma rays damages. Compared the protective effect of TFA on DNA strand breaks induced by hydrogen peroxide and gamma rays, the former was better. The protec-live mechanism of TFA on DNA strand break could be related to savenging free radicals.]]> Wang Deqing,Shen Wenmei,Tian Yaping and Jiang Cien 1948true 2+ supplementation in the bacterial growth media resulted in about 3-fold increasing of SOD activity. The special protein expressed accounts for 14% of the total protein of the bacteria.]]> Luo Xunyi,Zhou Ljun,Chen Xlaosul,WangYuxiao,Xie Bangtie,Wang Jingyi,Chen Yuping and He ling 1947true 7-MT and Cd7-MT were prepared respectively. The scavenging effects of Zn7-MT and Cd7-MT on hydroxyl radical were compared at various pH values, and at pH 6 the effects of Zn7-MT were compared with related proteins and inorganic zinc salt. From these results it is concluded that the ability of Zn7-MT to scavenge hydroxyl radical is far stronger than that of Cd7MT and that the ability of MT to scavenge hydroxyl radical derives from reduced -SH group in MT.]]> Yue Xinping,Zhou Jiehao and Cheng Shi 1946true E．Coli J5 strain were enriched and screened by panning and filter blot with E．Coli J5 strain from a human antibody library. Four positive clones binding to E．Coli J5 strain were determined by ELISA. The results of inhibition test showed that the binding of positive clones to antigens could be respectively inhibited by E．Coli J5 strain, E．Coli Rc-LPS and anti-CGL MAbs to E．Coli J5 strain. PCR amplification showed that all of the four positive clones contained the expected heavy and light chain genes of human Fab fragment. SDS-PAGE and Western blot analysis confirmed that positive clones were able to express about 50 ku proteins after induced with IPTG.These data indicate that four positive clones express the human Fab fragment with antigen specificities.]]> Yu Xiaofeng,Wang Haitao and Huang Ce 1945true Chen Sheng,Huang Zhiyue and Liu Yanru 1944true Hu Yingqing,Zhang Weisu,Zhu Aiping and Xia Zhiyin 1943true Trichosanthes Kirilowii Maxim has a highuracil-specific activity. In the absence of urea and under conditions of PH3.5, 50℃, it cleaves almost exclusively and uniformly at -NP ↓ U-. Inconjunction with RNase T1, U2 and limited alkaline hydrolysis, the RNase TCS is useful in direct enzymatic RNA sequence analysis.]]> Zhao Kun and Hua Ling 1942true Astragalus (TFA), total saponins of Astragalus (TSA) and total polysaccharide of Astragalus (TPA) on superoxide anion radical and hydroxyl radical have been studied by ESR method. The results showed that all the three principles have stronger scavenging effect on superoxide anion radical than on hydroxyl radical. The intensity of scavenging effect of the three principles was TFA＞TSA＞TPA. It is suggested that the TFA and TSA are the main component in Astragalus mongholicus Bunge for protection against free radical damages.]]> Wang Deqing,Shen Wenmei,Tian Yaping,Sun Zhongyi,Cong Jianbo and Wu Ke 1941true 2 against serum antigen of human breast cancer was reported.The results demonstrated that the antigen recognized by McAbGB2 is a complex protein which is composed of carbohydrate and protein and it is susceptible to heat. Competitive ELISA test showed that the binding of McAbGB2 to its antigen could not be inhibited by ferritin and CEA. Western blot revealed that McAbGB2 recognized antigen with molecular weight of 116 and 45 ku. The antigen recognized by McAbGB2 exists in breast cancer tissue and serum. ELISA test with McAbGB2 showed the rate of accord with positivity is 98 %(50/51) in breast cancer patient sera and the rates of false positivity are 6 % (3/50) in normal serum samples tend 6.7% (1/5)in benign breast disease sera respectively. The antigen recognized by McAbGB2 may be a new tumor-associated antigen.]]> Cai Guiying,Chen Hong and Wei Ling 1940true Qu Chengkui,Wei Handong,He Fuchu,Wang Lisheng and Wu Zuze 1939true Yang Tangbin and Mei Shangyun 1938true E.coli has been constructed. The vector contains glutathione S-transferase (GST) gene as the first cistron, followed successively with translational enhancer, SD sequence, stop codon, start codon and multiple restriction enzyme sites for cloning(MCS).3'-terminal framgents of human bone morphogenetic protein (hBMP)gene 2A and 3 were inserted into the MCS respectively. After induction, un fused hBMP2A and hBMP3 expressed and occupied 10% and 15% of the total cell protein respectively. The GST gene in the plasmids were further shortened from 660 by to 206 bp. The expression level of hBMP2A and hBMP3 were double by the plasmids containing short GST gene as compared to that of the corresponding plasmids with large GST gene.]]> Liu Xinping,Chen Sumin,Chen Nanchun,Zhao Zhongliang,Chai Yubo,Cui Youhong and Xue Yongtao 1937true IC50) are within 10-6～10-8mol·L-1. The rate constants (kq) of catalytic dismutation of superoxide radicals (O-·2) are in the range of 106～108mol-1·L·s-1.Simultaneously it has been observed that a several mimetic compounds of then have antineoplastic activities and increasing antifreezing effect of rices.]]> Liao Zhanru,Fu Huihua,Tian Tingliang,Cai Hanlin and Liu Wanqiao 1936true 125I-C1q showed a specific, dose-dependent,saturable and reversible binding involving specific membrane receptors on Jurkat cells, Goldstein analysis and Hill plot of C1q binding showed 1.1×106 binding sites per cell with an average affinity of 1.5×107mol-1 and a Hill number of 0.9643 at normal ionic strength(I=0 .15) and temperature(37℃). The experiment with the anti-Clq monoclonal antibody A4，which recognizes the collagen-like region (CLR) of C1q, established that it is via its CLR that C1q binds to Jurkat cell receptors. Western blotting with the anti-C1qR antibody 112 revealed that the C1qR on Jurkat cells is a membrane protein of 70 ku.]]> Chen Zhengliang and Xie Peirong 1935true Zhu Wuming,Jiang Linhua,Chen Shude,Sun Shufeng and Chen Jiasen 1934true Hong An,Yao Zhibin,Gu Yaoming and Chen Yici 1933true 229 induced by retinoic acid (RA) and 1, 25-dihydroxyvitamin D3 (1, 25 (OH)2VD3) both for two days and phorbol 12-myristate 13-acetate (PMA ) for six hours. The results showed that total PKC activity was markedly increased after treatment with three inducers (P< 0. 05 ). The cytosolic PKC activities were significantly increased in RA and 1, 25 (OH)2VD3 treated CCL229 cells (P< 0. 05 ). In PMA-treated cells，the membrane ratio(membrane activity/total activity) was significantly increased (P<0. 01).The levels of PKC inhibitor in cytosolic membrane supernatants after treatment with three inducers were all decreased. Among these three inducers，1，25 (OH)2VD3 caused significantly difference compared with control. The results implied that PMA translocates PKC from the cytosolic to membrane fraction，PKC and its inhibitor in RA，1，25 (OH)2VD3 and PMA-induced cells are in different relatively parralel relationship.]]> Wang Hongmei,Yu Bingzhi,Wang Yunqing and Song Jindan 1932true Liu Dailan,Qiang Boqin,Yuan Jiarigang,Li Yuan and Liang Zhiquan 1931true 2O2－Fe3+ system.It was shown that there was a significant dose-dependent relationship between concentration of H2O2－Fe3+ and the content.of DNAdsbs occuring in cells. Moreover, the DNA damage in terms of dsbs became heavier as the time of incubating cells with H2O2－Fe3+ was extended but could be inhibited by catalase considerably.The sensitivity of PFGE was estimated by the concentration of H2O2－Fe3+ which were 0.3 mmol/L and 6 pmol/L, respectively.]]> Sui Jianli,Lu Xing,Zhou Pingkun and Fang Yunzhong 1930true Ye Jianxin,Xiao Chengzu and Zhang Zhengguang 1929true 10 secreted monoclonal antibody against human D-dimer as a template, the cDNA fragment encoding the Vk domain was amplified by reverse transcription and polymerase chain reaction (RT-PCR) with a set of universe primers that were on the FR1 region and FR4 region of Vk domain respectively.The RT-PCR product was ligated with pUC18. The result obtained from the restriction enzyme fragments and DNA sequencing showed that the length of the Vk was 330 bp, and the distinctive structures of the immunoglobin CDR1，CDR2 and CDR3 bf Vk gene appeared.]]> Guan Xiaoqun,Zhang Yaming,Li Yong,Ding Hao,Zhu Yunsong and Song Houyan 1928true 1-AgTx), a presynaptic neurotoxin obtained from the venom of snake Agkistrodon blomhoffii brevicaudus (Jiangshu and Zhejiang Province, China) was further purified by the method of preparative electrophoresis. The polymerism was found in the native component of β1-AgTx and the purified β1-AgTx-OH- which was solubilized in the slight basic solution (pH 7.5) by SDS-PAGE analysis, however, the amount of polymers seems to be stable observed in the different days. Moreover, by two-dimensional electrophoresis analysis, it was found that both components of β1-AgTx and β1-AgTx-OH- were composed of three isoforms (pI 7.0, 5.8 and 5.4) with different contents, and the rate of content of isoforms in the component of β1-AgTx-OH- was changeable observed in the different days, but not in the β1-AgTx.]]> REN Huimin and JI Yonghua 1927true RPL promoters regulated by temperature sensitive repressors. SDS-PAGE analysis demon-strated that the enzyme was produced to the extent of as much as 38% of the total cellular protein. The soluble recombinant enzyme can be released from bacterial cells and enriched to purity of more than 85% by the Johnson's freeze/thaw method. Mouse anti-rabbit muscle adenylate kinase monoclonal antibody can strongly crossreact with the recombinant enzyme.]]> JING Enxuan,ZHOU Bo,LUO Jie and ZHANG Hongjie 1926true 3H］ DPDPE to opiate receptor. The enkephalin-αⅠ interferon fusion protein was shown to have more antiproliferative and antitumor activities against opiate receptor-bearing tumors, both in vivo and in vitro.These results suggest enkephalin-αⅠ interferon fusion protein may be able to target opiate receptors on tumor cells.]]> ZHU Qianzheng,WANG Jing,ZHANG Xiaobing,LI Mengfeng,ZHANG Ruli,CHEN Wanqiu and HOU Yunde 1925true in vitro site-directed mutagenesis using the double primer method and then expressed in E.coli K38. The characteristic analysis of the enzymes showed that thermostability of GIK253RA198C was significantly lower as compared with wild-type GI in 80℃. Moreover, its optimum reaction temperature decreased from 75℃ to 70℃. The results are explained by the kinetic parameters. Previous experiment indicated that the same mutation of K253R produced different effects on the thermostablity of SM33 GI and A.missouriensis GI. It is clarified here, based on structure and mechanism, that the result was due to the minor difference between Lys253's positions in the two GI structures.]]> WU Chuanjin,WANG Chen,TENG Maikun,WANG Chun,XIAO Yazhong,WANG Yuzhen,NIU Liwen and CUI Tao 1924true 3H］DL-glutamate binding to rat cortical synaptosomal membranes were studied by radiolabeled receptor assay. O -·2 results in a marked increase of specific glutamate binding. EBSELEN (1 μmol/L ) shows an inhibition effect on glutamate binding influenced by O -·2.]]> YI Yong,YANG Xiangliang,ZHAO Xilong,ZHANG Hengshan,QIN Yuhui and XU Huibi 1923true C3 has never been revealed in the Han population before. The Gene frequencies are as follows: TfC1=0.7420, TfC2=0.2420, TfC3=0.0027, TfDchi=0.0133. There is a good agreement between the observed and expected values corresponding to the Hardy-Weinberg equilibrium. The allele frequencies for TfC1, TfC2 and TfDchi are quite similar to those previously reported.]]> GUO Yaojun,LI Xinhui,ZHU Haizhen and SHEN Ziwei 1922true Candida utilis, a major structural cell wall protein with molecular mass of 33 ku was found.The 33 ku protein was quite different from most of the other cell wall proteins in Candida utilis. In cell wall it was not hydrolysed by trypsin, but was sensitive to subtilisin. The 33 ku protein was found to exist in cell wall of all logarithmic growth phases. Especially in early logarithmic growth phase, it was proved to be the only protein that was not sensitive to trypsin. The result demonstrated that this protein was important for connection of cell wall skeleton components-glucan, and for the intact structure of cell wall. It was an important integral cell wall protein.]]> ZHANG Xiping,ZHANG Chuyu and WANG Esheng 1921true Rana pipiens. β-Galactosidase in retina neurons was still identified in two weeks after transfection. Gene transfection to retina neurons is non-selectivity for any neuron. The transfected neurons are around the injection area. Comparing with the normal, the neurons transfected in retina of frog optic nerve transected distribute in a slightly wider area.]]> XIA Lingchao,LI Jinzhao,CHEN Yan,QIU Rong,ZHANG Ying and DENG Wei 1920true YAO Baoli,XU Dalun,HOU Xun,HU Kunsheng and WANG Aojin 1919true WU Xiaohua,LIU Wangyi,XU Lei and LI Minqian 1918true ZHAO Jiquan,ZHAO Jingquan,YANG Zixuan,ZHANG Jianping and JIANG Lijin 1917true in vitro, The inhibitory effect of L-NNA on the synthesis nitric oxide (NO) was investigated, which was induced by IFN-γ in macrophages. It was found that L-NNA could inhibit the synthesis of NO, the inhibition was related to L-NNA dose, and L-arginine could reverse the inhibition. The results suggest that L-NNA likely competitively combined with the activate site of inducible nitric oxide synthase (iNOS), that inhibited the synthesis of NO from L-arginine pathway in macrophage.]]> NIU Yuxin,CHU Yi,YANG Dianer and LI Huizhu 1916true GU Shanqing,LIANG Yunyan,WANG Yuemin,LI Baoyuan,WANG Jun,MA Yan and WANG Daishu 1915true 4 were used as template protein to predict the 3D-structure of the functional domain (residue 106～322) of hIL-6R. The characteristics of the conservative structure-conformation in CBD of hIL-6R were described. The model may provide for the interpretation of the results obtained with introducing site-directed mutation into soluble IL-6R and the 3D-quantitative analysis on the structure-function relationship of IL-6R functional domain.]]> SUN Tao,REN Yunfang,LU Zhongzheng,LI Song,JIAO Kefang and SHEN Beifen 1914true LIAO Lifu,HE Yuyuan,LIU Chuanxiang,YUAN Yali and LI Guirong 1913true E.coli and Bacillus subtilis. The recombinant plasmids were used to transform B.subtilis DB104，a mutant strain deficient in alkaline and neutral protease，then they were expressed. They were (M222A，E156S) and (M222A，E156S，V165I). The property analysis of these enzymes revealed that the Subtilisin E E156S substitution enhenced the hydrolysis Kcat/Km by 90% while keeping thermal stability and oxidation-resistance unchanged，however the V165I mutation reduced the Kcat/Km value.]]> CHEN Weidong,MA Jianhua and ZHU Liuqin 1912true YIN Yinliang,WANG Keqin and CHEN Baosheng 1911true LI Jinying,ZHAO Faji,GUO Junsheng,YE Xuting,HU Xiaojian,ZHANG Zhihong and ZHOU Hanqing 1910true in vitro, human umbilical vein endothelial cells were cultured and divided into four groups: control, HDL+LDL, apoAⅡ+LDL and LDL group, which were observed the morphological changes with phase-contrast and transmission electron microscope and measured the release of lacate dehydrogenase (LDH) and level of 6-keto-prostaglandin F1α (PGF1α). The endothelial cells injured by LDL showed cell contraction, increased release of LDH and decreased level of 6-keto-PGF1α. However, normal morphology and LDH release as well as PGI2 synthesis in endothelial cells were found when HDL or apoAⅡ was added to culture media before LDL injury. The results indicated that both HDL and apoAⅡ could resist the injurious effect of LDL on cultured endothelial cells.]]> LI Jian,JIANG Lei and LIU Qinghua 1909true in situ PCR in cultured human small intestine cancer metastastic ascites cell line cells. The results show that in situ PCR is much more sensitive than in situ hybridization.]]> MA Qi,ZHANG Xiyuan,LIU Ting,YANG Jianqi and XU Yaoxian 1908true QIU Zhaohua and HE Fuchu 1907true 2+-ATPase proteoliposomes were reconstituted with ganglioside GM3 and soybean phospholipid by addition of detergent, sonication and chromatography. It was observed that GM3 may enhance the activity of Ca2+-ATPase in proteoliposomes and increase the diameter of uniform protein particles in sealed-well proteoliposomes vesicles which was evidenced by the negative-staining microscopy and freeze-fracture replica.]]> WANG Lihua and YANG Xiaoyi 1906true 2+. The results suggested that the RF15 may be a new regulatory sequence in the PSA promoter region. The regulatory protein, which binds to the RF15 sequence, may enhance the androgen induction via interaction with androgen receptor.]]> ZHANG Jianye,PANG Weiqiu,WANG Xin,ZHANG Lianying and RU Binggen 1905true YANG Ming,ZHANG Zhiwen and HOU Weimin 1904true 60-phosphatidylcholine liposome (C60-concentration as 20 mg/L) and HeLa cells and illumination with the Tungsten-Halogen lamp (4 000 lx, 30 min), the most of cells were killed by the evaluation of MTT assay (3-(4,5-dimethythiazol-2-yl)-2,5-diphenyl tetrazolium bromide). Biochemical test showed that photoexcited C60 led to the decrease of the sulfhydryl content of the membrane protein and the increase of the malonidaldehyde (MDA) with the peroxidation of the membrane lipid. SDS-PAGE showed the conjugation of the membrane protein. Fluorescence polarization (FP) indicated that the fluidity of the cell membrane increase.]]> QIAN Kaixian,YAN Qingfeng,HUANG Wendong and LI Wenzhu 1903true LI Aihua,HUANG Rubin,YANG Dianer and JIN Youyu 1902true -6 mol/L and 4×10-5 mol/L, and cocultured for 24 hours. Ladders of DNA electrophoresis and hypodiploid peaks were discovered in the two treated groups. Compared with the control groups, the cell cycle percentage of S in treated groups by SS was much increased, but the percentage of G1 was significantly decreased (P＜0.05), and the percentage of G2 in ST treated groups was much increased, but both the percentage of S and G1 were much decreased (P＜0.01). The well-distributed chromatin DNA was seen in untreated cells, but fragmentary DNA in treated cells. The granuliform microfilament regularly lined along and formed the intact cell morphology in untreated group cells. The microflament clustered in disorder and formed clusters in induced cells. The results showed that the cell cycle and cytoskeleton may contribute to the CNE-2Z cells apoptosis induced by the inhibitors of protein kinase C.]]> HE Zhiwei,CHEN Nanyue,LIAO Xinbo and CAI Kangrong 1901true Schistosoma japonicum 26K antigen(Sj26GST) into Escherichia coli-Mycobacteria shuttle plasmid pBCG-2000 and their expression efficiency were investigated in Mycobacterium smegmatis. The plasmid which contains promoter of human Mycobacterium tuberculosis heat shock protein 70(hsp70) was digested with NcoⅠ and modified with two different ways to lead to two kinds of SD squences, and then ligated with Sj26GST encoding gene. The DNA fragment contained hsp70 promoter and Sj26GST gene was cloned into pBCG-2000, and finally four recombinant mycobacterial expression vectors that are different in SD sequence, orientation and copy number were selected. The expressed native recombinant Sj26GST(rSj26GST) could be observed on SDS-PAGE about at the molecular weight of 26 ku obviously. Analysis with protein density scanning indicated that the expression efficiency that containing double-copy promoter-foreign gene vector was the highest and the expressed protein was about 1.6 folds than that of others. The cloning direction and SD sequence had no significant effect on expression efficiency.]]> CHENG Jizhong,HUANGFU Yongmu and HAI Tao 1900true 2+ and Cu2+) and different elution strategies (pH and imidazole gradient) were compared. The results showed that Cu2+ were more effective than Zn2+ in retaining the His tagged Fab protein. pH gradient showed better consistency in Fab recovery than imidazole gradient. Using Cu2+ IMAC more than 95% pure rh Fab could be obtained by one step purification.]]> ZHU Yingchun,WANG Yan,LIU Qunying,HUA Bing and GAO Rongkai 1899true LIU Shangxi,CHEN Yuan,ZHOU Mei and SUN Manji 1898true H(plate height) of two kinds of CCPC(centrifuge countercurrent partition chromatography) columns. The results of both theoretical deriving and experiments reveal that the diameter of column, capacity factor, mass transfer coefficient, the average diameter and velocity of mobile phase droplets are main factors that determines H. Mathematics analysis is given.]]> ZHAO Yonghong and KIM Lock Song 1897true Km and Vmax were determined. Km(NO)and Km(NADH) were 0.128 mmol/L and 0.208 mmol/L respectively. Vmax for N2O was 11 363 min-1. The spectroscopic study indicated that: rF·P-450nor has the typical characteristics of a heme protein, and has the maximum absorbance at 413 nm. After added Na2S2O4，the maximum absorption peak was moved to 405 nm. After added CO and Na2S2O4, the absorption peak was at 450 nm. When combined with NO, the absorption peak was moved to 430 nm. These spectrum characteristics were the same as those of the native F·P-450nor.]]> LIU Deli and Hirofumi Shoun 1896true 5-Ce(Ⅲ) ［BLMA5-Ce(Ⅲ)］. In additon, there were other products to the four mono-nucleotides (5-dAMP, 5-dTMP, 5-dCMP, 5-dGMP). It is suggested that BLMA5-Ce(Ⅲ) cleaves phosphooryl diester bonds along the direction of 5′～3′ at specific sides of double-chain DNA forming more exposed 3′-OH ends.]]> HUANG Deying,YANG Ming and WANG Kui 1895true SHEN Xianrong,JIA Fuxing,WANG Ling and LEI Chengxiang 1894true in vitro is reported. The tests showed that: the purified antibacterial peptide CM4 component can kill human myelogenous leukemia cancer (K562). The observation of ultrastructure of the cancer treated with the antibacterial peptides by using the scanning and transmission electron microscope and the microfault picture analyses of laser confocal microscope. The results showed that the pruified antibacterial peptides brought about a series of pathological changes on cancer cells. As a result, the cells were high swelling, plasmolysis and disorders in subcellular and membrane structrue occurred, outer coat of cell dissolved, thus appearing irregular holes opening in a utensil and serious broken of skeleton structure of the cells. The membrane structure of some areas was broken and injured and intracellular substances leaked out. As a result, the cells disintegrated into small fragmants and died.]]> ZHANG Shuangquan,JIA Hongwu and DAI Zhuying 1893true 25/16A3-F) and human monocytoid leukemic cells by oral administration at one dose of 100 mg/kg.The CGS and DGS were more effective than Ge-132 in enhancing MTC effect at the above dose,and CGS had the strongest effect. The Mφs activated in vivo by CGS, DGS and Ge-132 at one dose of 100 mg/kg showed increased incorporation of ［3H］ choline into phosphatidylcholine (PC), and the most significant increase was observed when Mφs were activated by DGS. Mφs activated in vivo by Ge-132 also showed increased incorporation of ［32P］Pi and ［3H］choline into PC, and decreased incorporation of ［32P］Pi and ［3H］inositol into PI when compared with resident peritoneal Mφs. No significant difference was observed on the incorporation of ［32P］Pi and ［3H］inositol into polyphosphoinositide (PIP and PIP2） between the Ge-132 activated Mφs and the resident peritoneal Mφs. The enhanced PC turnover of Mφs might be necessary for the expression of MTC in activated Mφs.]]> WANG Yin,LIU Yan and CUI Zhaochun 1892true GU Yongjun,MA Jiabao,Fumitaka Oyama and Yasuo Ihara 1891true SUN Juan,CHEN Yuan,ZHOU Mei,GE Zhongliang and LI Mingtao 1890true SHAN Weisong,ZHANG Guorong,ZHANG Yuehua,LIANG Yingwu,LI Chunying and WU Xiru 1889true in vitro. The results showed that along with the increasing of selenite concentration, α A-crystallin transcription decreased, then went up sharply at 5×10－5 mol/L Na2SeO3 . The results suggested that α A-crystallin might at least respond to high concentration of selenite and express as a stress protein. But β23-crystallin gene transcription showed increasing followed by decreasing with the increase of selenite concentration. It indicated that selenium might play a critical role in lens epithelial cell differentiation.]]> WANG Bo,HE Haiying,JIA Weihong,ZHANG Jiaping,LIANG Kang and ZHANG Changying 1888true ex 284 and 276, λem 390 and 386 nm respectively), but Trp, Pro, His are long (λex 290, 308 and 320 nm, respectively). The phosphorescence lifetime of Phe, Trp, are the longest (about 7 s), Tyr, Pro and His are short (2.84,1.31 and 0.49 s respectively). The phosphorescence spectra of amino acids in different alcohol (methanol, ethanol, n-propanol, n-butanol) are changed little, but the lifetime becomes shorter as the polarity of alcohol decreasing. The effect of pH on the spectra of amino acids were studied. The stokes energy loss and the excited pK*a were also measured.]]> HUANG Ruheng 1887true YU Min-Zhong and WU De-Zheng 1886true nHSA) and galactosyl poly-L-glutamic acid (GalnPLGA), were synthesized chemically. IC50 of synthetic ligands inhibiting the binding of 125I-asialofetuin (125I-ASF) to ASGPR of rat liver cell membrane were assayed. The results showed that Gal12HSA, Gal15HSA, Gal26HSA, Gal30HSA and Gal34-PLGA could all efficiently inhibit the binding of 125I-ASF to ASGPR. The affinities of GalnHSA increased along with the increase of the coupling number of galactosyl. Here, the synthesis and the affinity identification of Gal34PLGA with ASGPR were firstly reported. These synthetic ligands with abundant sources and simple preparations may be used as targeting ligands for the liver-targeting delivery of drugs or genes.]]> LI Chong-Hui,WEN Shou-Ming,CHI Mu-Gen and SUN Man-Ji 1885true E.coli, and visualized by SDS-PAGE analysis.]]> CHEN Yan,ZHANG Ying,LI Jin-Zhao,DENG Wei,XIA Ling-Chao and QIU Rong 1884true Candida cylindracea in isooctane/octanol system. The results demonstrated that the salt hydrate pair —— Na2SO4·10H2O/Na2SO4 had the capability of bufferring water. The sensibility of enzyme to water was decreased by adsorbing the lipase on celite —— an apolar carrier. In addition, Esterification could be improved effectively by adding molecular seive to remove the byproduct-water.]]> CUI Yu-Min,WEI Dong-Zhi and YU Jun-Tang 1883true LIU Chun-yu,ZHANG Chun-ling and XIA Jia-hui 1882true 2+ concentration(［Ca2+］i) after the action of verapamil was investigated under different extracellular calcium concentrations. And the research about the effect of verapamil on experimental silicosis rats were conducted. The results showed: the ［Ca2+］i rise induced by α-quartz was caused by enormous extracellular Ca2+ influx and the overloading of the intracellular Ca2+, which directly led to the death of alveolar macrophages and further the formation of silicosis. Verapamil could inhibit the ［Ca2+］i rise, decline the death of cells. The results of the research about experimental silicosis indicated: verapamil has apparent inhibitory effect on the starting of silicosis. It suggested that verapamil appears to be a prevention and cure drug of silicosis.]]> ZHU Wei-hua,WANG Jin-xi,LI Hai-shan and CHEN Yao 1881true TK gene under the control of CEA promoter was constructed (AdCEATK). Titer of the purified recombinant adenovirus is about 1012pfu/ml. HeLa cell (CEA-negative cell) infected with AdCMVTK became sensitive to GCV, while HeLa cell infected with AdCEATK was not. On the contrary, the LoVo cell infected with both AdCMVTK and AdCEATK were sensitive to GCV. It was shown that CEA promoter has a good tumor-specificity. Significant bystander effect was observed in the AdCEATK/GCV system too. This system should be useful for tumor-specificity suicide gene therapy of CEA-positive tumors.]]> XU De-hua,GE Kai,JIANG Qiong,ZHENG Zhong-cheng and LIU Xin-yuan 1880true 1,ribozymeC2) were designed and synthesized according to the secondary structure of HCV-RNA 5′-untranslated region and part of the neighbour C-region. Firstly, the cleavage of the four Rzs were tested in vitro, and only the ribozyme with GTA↓ motif at-11nt site of HCV-RNA showed cleavage activity. RzA-RNA and the combinated pCl-neo-luciferase in which a luciferase gene were ligated downstream the target sequence were then co-transfected into HepG2 cell lines with lipofectine. The cleavage of RzA-RNA was tested by determined the expression of luciferase gene. Therefore, the gene of RzA was ligated into expression vector pCl-neo. This pCl-neo-RzA and the vector pCl-neo-luciferase were co-transfected into HepG2 cell lines again with lipofectine. Since the pCl-neo-RzA was more stable than RzA-RNA in vivo and could produce RzA-RNA continously, it showed better cleavage activity.]]> LIU Li-zhong,WANG Sheng-qi,ZHU Bao-zhen and SUN Zhi-xian 1879true LI Qin,LI Li,WANG Zhao-jie and QIAN Shi-jun 1878true LI Li,LI Qin,WANG Zhao-jie,YE Jun and QIAN Shi-jun 1877true 2+ and protein kinase C in platelet aggregation, suspensions of aspirin-treated, 32P-prelabled, washed pig platelets containing ADP scavenger in the buffer were stimulated by Ca2+ ionophore A23187 and PMA,a stimulator of protein kinase C. The results indicated that: (1) 1～20 μmol/L A23187 induced platelet aggregation,as well as the phosphorylation of 40 ku and 20 ku proteins.There were dose-respone and time-respone effects of the protein phosphorylation in A23187-induced platelet activation. (2) A23187 and PMA were synergistic in platelet aggregation and protein phosphorylation. (3)Stauroporine, a protein kinase C inhibitor, in concentration of 1 μmol/L,largely suppressed platelet aggregation and completely suppressed phosphorylation of 40 ku and 20 ku proteins induced by 20 μmol/L A23187. The results imply that Ca2+ mobilization alone could activate protein kinase C in platelet, and Ca2+-induced platelet aggregation is largely dependent on activation of protein kinase C.]]> CHEN Ri-yan,JIANG Li-ming,QIN Yan-mei and LIANG Nian-ci 1876true TM, and then tested the expression level of CAT gene with DIG-labeled anti-CAT ELISA. The results indicate that the first intronic +544～+855 segment of COLⅠA1 gene has an enhancing effect on the expression of CAT gene driven by the promoter in the tranfected human fetal tendon fibroblasts and Tca 8113 tongue cancer cells. Interestingly, the first intronic +820～+1 093 segment has strongly stimulating effect on the expression of the marker gene in the tongue cancer cells, but inhibitory effect in the fibroblasts.]]> CHEN Jun-jie,YANG Shao-hua,WANG Ruo-han,PENG Wen-zhen,MA Ying-hong,LIU Zhi-min and LI Chang-long 1875true DAI Yan and CHEN Wei-Feng 1874true SUN Juan,CHEN Yuan and ZHOU Mei 1873true 3TG2TGT2G5TG2TGT(CP1) and core promoter(Cp) fragment of hepatitis B virus(HBV) had a good specificity and stability, which was demonstrated by electrophoretic mobility shift analysis and DNaseⅠ footprinting experiment. Gel retardation assay showed CP1 could inhibit a specific cellular factor binding to Cp fragment in rat liver nuclear extracts. No inhibition of factor binding was observed by oligodeoxyribonucleotide CP3(TGTG2TG5T2GTG2TG3), which could not form triplex with Cp fragment. These results indicate that specific repression of gene transcription of HBV DNA may be possible by triplex-formation.]]> LIU Ding-xie and WANG Chang-cai 1872true ZHANG Xiao-wei,WANG Fu-shan and TONG Tan-jun 1871true Pichia pastoris heterologous gene expression system was utilized to produce attractive levels of a variety of intracellular and extracellular proteins of interest. Lateolabrox japonicus growth hormone gene was cloned into the yeast integrative vector pHIL-D2, which was then transformed into his4 mutant yeast GS115. PCR fast detection methods was performed to screen the positive transformants and dot-blotting was used to screen the multiple-copy transformants. Via inducing AOX1 promoter by methanol, growth hormone could be expressed intracellularly. SDS-PAGE and Western blot were applied to confirm the product.]]> CHEN Dan,YANG Feng,WANG Wei and XU Xun 1870true in vitro as an internal standard, and then it was done RT-PCR procedure together with cellular cDNA. The two kinds of amplified cDNA fragments could be distinguished after agarose gel electrophoresis, because there were difference in their length. The concentration of cellular MDR1 mRNA was derided from the ratio between the intensities of the bands corresponding to the amplified products. The test for characterizing the MDR1 expression offers high sensitivity and specificity and is therefore of great clinical relevance.]]> LI Yong and WANG Yu-zhi 1869true 32P］ ATP labeled CREB and NF-κB consensus sequences was found to be increased 4 h after WBI with 75 mGy X-rays, being 7 and 5 times higher, respectively, than that of the sham-irradiated control. Competitive inhibition analysis with excessive amount of unlabeled consensus sequences completely blocked the binding in corresponding EMSA, demonstrating the specificity of the reaction. The results suggest that low dose WBI could selectively activate the transcription factors CREB and NF-κB, which would induce specific gene transcription in the splenic cells leading to their functional activation.]]> CHEN Sha-li and LIU Shu-zheng 1868true E.coli by using GST expression system. The results showed that the bacterially expressed CPP32 protein is auto-cleaved and capable of cleaving in vitro-translated PARP, thus is fully functional.]]> CHEN Ya-bing,YU Chun-dong,GUO Ben-chang,DING Mei,GUO Shu-zhen,Zeng Ding and WEN Long-ping 1867true 4, after which amyloglucosidase is attached to it. The optimal reaction conditions and the kinetics of the immobilized amyloglucosidase are determined and compared with those of the free enzyme. The activity of the immobilized amyloglucosidase shows no decay after 10 batches of starch hydrolyses, the total reaction time of which is more than 24 h at 55℃.]]> QU Hong-bo,CONG Wei,WEI Xin-gui,CHEN Jian-fei and OUYANG Fan 1866true o moiety of mitochondrial F1Fo can be quenched by the addition of Hypocrellin B. The determination of the Stern-Volmer plot at different temperatures is carried out. The result shows that the Ksv increased with the increase of temperature. The experimental result of the time-resolved fluorescence decay shows the decrease of lifetime of tryptophan fluorescence of Fo with the increase of concentration of HB. No shift in the absorbance spectra of F1Fo were occurrent at varying concentration of HB. The results support the dynamic quenching. In addition, HB possess the necessary characters to be used as a quencher in the hydrophobic phase as follows: the low concentration of effective quenching; no effect on the activity of F1Fo; the ratio of the partition coefficients between lipid-phase and water-phase as high as 16 560∶1. So HB can be used as a ideal fluorescence quencher for the study of conformational change of Fo moiety of membrane-bound F1Fo complex.]]> LI Sheng-guang,FENG Zhao-yang,YUE Jia-chang,XU Ting and LIN Zhi-huan 1865true HU Qing-liu,DING Zhen-hua,TAN Xiao-hua and WANG Hong 1864true E.coli. A highly expressed vector of MK was constructed.]]> XUE Yong-tao,HUANG Wei-jin,SHI Bin,CHEN Bin-fu and JU Gong 1863true kαβ gene transfer on the growth of the transferred mouse tumor cells in vivo, the α and β chain cDNA were synthesized, and inserted into a retroviral vector, to construct two Ⅰ-Ak expressing recombinant plasmids pLSXN-Akα and pLSXN-Akβ. By means of a liposome-mediated gene transfer procedure, these two recombinant plasmids together were introduced into mouse lymphoma EL4 cells and mastocytoma P815 cells. The expression of the Ⅰ-Akαβ protein on the surface of the transconducted tumor cells was tested by FACS using Anti-Ak-FITC. Afterwards these tumor cells were injected subcutaneously into autologous mice C57BL/6(H-2b) and DBA/2(H-2d) respectively. It was observed that tumors developed at the beginning within the mouse bodies, and disappeared after a few weeks with injection. While the nontransferred tumor cells grew continuously. These results show that the tumor immune response can be stimulated by the tumor cells transferred with allogenic MHC class Ⅱ gene, in the absence of B7 co-stimulatory signals.]]> DING Guang-zhi,TIAN Jun,LIU Yuan-lin,ZHANG Yang-pei and MAO Ning 1862true WANG Fang,ZHANG Shuang-quan and DAI Zhu-ying 1861true in situ hybridization method (ISHM) was set up with rat hippocampus slides as positive controls. The change of GAP-43 mRNA levels in the vestibular nucleus area were investigated by ISHM in the labyrinthectomy rats at 5、12、20 and 30 days after the operation. The results demonstrated that labyrinthectomy increased GAP-43 mRNA levels. The application of ISHM laid a foundation for the research of regenerative sprouting, synaptic remodeling and neuroplasticity in the vestibular compensation.]]> YU Rui-Yuan,MU Xiao-Dong,WANG Yan-Feng,SUN Jiu-Rong and XU Chang-Fa 1860true WANG Wen-Gong and TONG Tan-Jun 1859true n=2,P＞0.05).]]> LIAN Jian-Qi,ZHOU Yong-Xing and JIN You-Xin 1858true γ,was constructed by DNA recombination and transfected into C3H10T1/2 cells by gene transfection. The NCP4 cells stably expressing PKCγ was isolated successfully by Southern blot,Western blot and PKC activity assay.The NCP4 cells displayed an enhanced growth rate,especially under low serum conditions, apparently reduced dependence on serum. By using FCM, it was shown that NCP4 cells exhibited decreased percentage in G1 phase, increased percentage in S phase and G2+M phase. In contrast to control cells, NCP4 cells overexpressing PKCγ decreased anchorage dependence and formed small colonies in soft agar. Furthermore, it was observed that the expression of oncogene c-sis increased obviously in NCP4 cells, it may be one of the molecular mechanisms of the reduced dependence on serum in NCP4 cells. The results indicated that specific elevation of the PKCγ level directly affected the increase of growth rate and lead to some transforming phenotypes in C3H10T1/2 cells.]]> ZHAO Xia and LIU Hui-Tu 1857true 32P-labeled τ peptides were purified by a sequential FeCl3 micro affinity column and a C18 reverse phase high performance liquid chromatography. The results from a combined techniques of high voltage electrophoresis, manual Edman degration and auto gas phase amino acid sequence analysis demonstrated that the phosphorylation sites of τ (pretreated with PKA) by GSK-3 were Ser(serine)-195,Ser-198,Ser-199,Ser-202,Ser-235,Ser-262,Ser-356,Ser-404, Thr(threonine)-205 and Thr-231. Among them, Ser-198,Ser-199,Ser-202,Ser-235,Ser-262,Ser-404,Thr-205 and Thr-231 are abnormal phosphorylation sites of τ found in Alzheimer disease. As the above phosphorylation potently inhibited the biological activity of τ, it was concluded that the phosphorylation of τ at above mentioned Alzheimer sites is critical to the inhibition of its biofunction, and that the interaction of PKA and GSK-3 might be a potential system responsible for the neurofibrillary degeneration seen in Alzheimer disease.]]> WANG Jian-Zhi,GRUNDKE-IQBAL I,SMITH A and IQBAL K 1856true Bmax of LDL receptor was significantly lower in As rabbit than that in normal rabbit(P＜0.01), while the value of Kd showed no difference; in HDL-treated rabbits, the value of Bmax increased significantly compared with the rabbits without HDL treatment(P＜0.01), while the value of Kd showed no difference yet. The results suggested that human plasma HDL could enhance the activity of LDL receptors on liver plasma membranes of As rabbit.]]> WU Xin-Wei,FU Ming-De,LAN De-Bin,DENG Ping and ZHOU Jian-Jun 1855true in situ hybridization and sequencing of the recombinant plasmid were used to determine the characteristics of differentially displayed cDNA fragment. The results indicate that it contains 170 base pairs and the incidence of homology with known genes is less than 80%, it was located in the renal tubule of SHR and no positive signal was found in the kidney of WKY.]]> LI Xin-Bo,ZHU Yi-Chun and YAO Tai 1854true Pichia pastoris, the cDNA coding for mature HRP isozyme C was subcloned into the vector pPIC9. After transformation, the recombinant P.pastoris strain which secreted nonglycosylated HRP(about 30 ku) and hyperglycosylated HRP (about 100 ku) was obtained. Through optimization of the growth conditions, the hyperglycosylated HRP as the sole product was secreted into fermentation broth and reached a level of approximately 4～6 mg per ml of culture medium. Remarkably, a certain activity of 2.24 U/ml was determined in the fermentation broth and showed the maximum absorbance at 403 nm.]]> JIANG Tai-Jiao,JI Xin-Song,ZHANG Ru-An,HUANG Yan-Hong,WU Xiang-Fu and YUAN Zhong-Yi 1853true LI Jun-Feng and WU Qi-Jiu 1852true 32P isotype. The cDNA probes were hybrided to high density cDNA microarray GF200 with 5 184 genes and ESTs,then the signals were analyzed by Pathways software. The results showed that the low density signals were distributed in three types of tissues and the number of genes and ESTs with the density value over 200 was respectively 110,134 and 158 among NPC,LC and NP tissue. The results indicated that the different expression genes existed among NPC, LC and NP tissues and some new genes could play an important role in NPC. The high-density microarray was a rapid and effective method to select the different expression genes.]]> HE Zhi-Wei,XU Liang-Guo,REN Cai-Ping,XIE Lu,GAN Run-Liang and YAO Kai-Tai 1851true Lü Xing,XING Rui-Yun,SUN Zhi-Xian,PEI Xue-Tao and WU Zu-Ze 1850true XIAN Hai-Qing,FAN Ming,WU Yan and QIU Zong-Yin 1849true A615／A280) reaches to 14 at pH 7.0,and it has been demonstrated by PAGE as a single band.]]> WANG Yong,QIAN Kai-Xian and DONG Qiang 1848true Kd and Bmax of rabbit liver plasma membrane HDL receptors were (7.17±1.18)mg/L and (622.5±146.1)mg/L respectively (n=7).]]> JI Yan,FU Ming-De,WU Xin-Wei and LIU Bing-Wen 1847true JIANG Yong,HAN Jia-Huai and GU Jun 1846true H.molitrix and A.nobilis. Besides establishment of species-specific RAPD electrophoretogram, some kinds of primers which can produce molecular genetic markers of RAPD with individuality- or population-specificity of these fish were found. A modified method to extracting genomic DNA from peripheral blood cells of fish, and the application prospects of RAPD technique in genetic diversity and variation of fish, and in fishery management and the evaluation of fish resources were discussed.]]> ZHANG Xi-Yuan,YANG Jian-Qi,ZHANG De-Chun,DENG Feng-Jiao,YU Lai-Ning and FANG Yao-Lin 1845true Km value of PI kinase for the utilization of PtdIns is 6.6×10-7 mol/L and for ATP 7.9×10-7 mol/L. In addition adenosine was found to be a strong inhibitor, Enzymatic activity was found to be stimulated by Triton X-100.]]> YUE Jia-Chang and GAI Li-Yun 1844true 53 gene and Rb gene, a P53 gene recombinant adenovirus vector, AdCMV P53, and a Rb gene recombinant adenovirus vector, AdCMV Rb, were transfected into the cultured vascular smooth muscle cells derived from human umbilical artery. The level of c-fos、c-jun mRNA was quantified by reverse transcription-polymerase chain reaction (RT-PCR). The content of c-Fos and c-Jun protein were detected by immunochemical staining. The senescent cells were characterized by β-galactosidase staining. Agarose gel electrophoresis of DNA was used for analysis of apoptosis. The results showed that introduction of wild-type P53 could increase the level of c-fos、 c-jun mRNA and protein, inducing the apoptosis of vascular smooth muscle cells. The introduction of exogenous Rb gene could induce cell senescence and down regulate c-fos gene expression. However, c-jun expression in vascular smooth muscle cells remained constant following transfection with AdCMV Rb.]]> WANG Shu,TANG Wei-Qing and LI Jian 1843true 2.]]> HUANG Jian,LI Wen-Xin,GOU De-Ming and JIANG Da-He 1842true I-V in NCNs was more significant(r=0.96) and the rectification insignificant;when the polarization current larger than 1.0 nA, the rectification occurred on both ends of the curve and then I-V curve showed S-type. The rectification of somatic NCNs was much significant than non-nociceptive ones(NNCNs). Rm,τ and Cm of the NCNs were evidently larger than those of NNCNs(P＜0.01 or P＜0.05).The materials obtained indicated that morphology and structural of cell membrane,volume of the soma bettween NCNs and NNCNs might have meaningful differences. It also means that physiological functions were unlike. The specificity of the electrical parameters in cell membrane might provide experimental materials for specific theory of pain sensation. Reporting to investigate electrical properties(Rm,τ and Cm) of the membrane in primary cortial nociceptive neurons had not found in literatures.]]> ZHANG Ri-Hui and TENG Guo-Xi 1841true ZHANG Jing and LIU Ci-Quan 1840true 2+］i transients were induced in porcine polymorphonuclear leukocytes in response to ATP and ADP, while no evident change in ［Ca2+］i was observed after AMP addition. The cells showed varied dose dependency upon ATP and ADP. In the Ca2+-free bath solution, ATP and ADP could still cause ［Ca2+］i elevation in the cells. The results showed that the P2 receptors are present on porcine polymorphonuclear leukocytes and should belong to the P2Y subclass of purinoceptor family.]]> CHEN She,NIE Yu-Sheng and HU Kun-Sheng 1839true E.coli at similar level. Correction of any one of the V region genes could partially resume the antigen binding activity. This result indicated that PCR primers introduced Vκ and Fd N terminal changes may seriously affect the antigen binding activity.]]> LI Jing,WANG Yan,WANG Zhuo-Zhi,LIU Qun-Ying,HUA Bing,CHEN Yu-Ping,ZHU Ying-Chun and DONG Zhi-Wei 1838true WANG Jian-Zhi,WANG Qun,WU Qiong-Li,I.GRUNDKE-IQBAL and K.IQBAL 1837true XIA Zhen-Wei,LI Yun-Zhu,CHEN Shun-Nian,SHEN Qing-Xiang,SHAO Jie,WANG Jian and YU Shan-Chang 1836true LI Ping-Yang,WANG Jiang-Cheng,TAN Run-Chu,LI Yong-Long,HE Qi-Cai and HUANG Yao-Xiong 1835true 125I-EGF on plasma membrance EGF receptor and radio-receptor assay when 125I-EGF was incubated with human glioblastoma multiforme cells were carried out. The results showed that there is a high affinity binding site on BT325 cells plasma membrance, exogenous GM3 could not alter the affinity (Kd) of EGF binding with cell-surface receptors, but decrease the receptor number. GM3 could significantly prolong the endocytosis process of EGF-EGFR complex. The concentration of EGF in culture medium was unchanged by GM3 and BBG,but that of EGF in cytoplasm was increased by GM3. It suggested that GM3 might inhibit the secretion of EGF.]]> LI Ai-Hua,YANG Dian-Er,CHEN Hong and HUANG Ru-Bin 1834true 2O2 and their autofluorescence directly, in order to study the effect of H2O2 on the antigenicity and the production of fluorescent product of LPO of RBC. The results showed that the changes of both antigenicity and autofluorescence were dependent on the H2O2 concentration and the time of RBCs exposed to H2O2, and the change of antiginicity was more susceptible to H2O2 than that of autofluorescence. At 102 fold lower H2O2 concentration than that made autofluorescence significantly increase, IgG bond to RBC. Furthermore, it was found that the changes of antigenicity and autofluorescence were related to forward scatter (FSC).]]> FU Nai-Yang and ZHANG Er-Xian 1833true Candida cylindracea. The transesterification reaction was apparently affected by the hydrophobicity of reaction medium, the water activity of the reaction system and the structure of the substrates. n-Octane was found to be the best medium for the reaction and the optimum water activity for the reaction is about 0.55.]]> ZONG Min-Hua,CHENG Qian and LIN Ying 1832true LU Yi-Fan,TIAN Chai,DENG Ji-Xian and HUANG Pei-Tang 1831true 1 368～373,α2 381～388) structures in p53 protein C-terminal region based on its mRNA secondary structure template and Chou-Fasman's protein secondary structure principle of prediction. The result was conformed by the other four methods of protein secondary structure prediction that are based on the multiple sequence alignment (accuracy=73.20%). Combine with the 31 amino acids crystal structure of the oligomerization, the three dimensional conformation of p53 C-terminal 108 residues was built using the SGI INDIGO2 computer. This structure further expounds the relationship among those biological function domains of p53 C-terminus at three-dimensional level.]]> ZHANG Yan and LIU Ci-Quan 1830true E.coli system. Moreover, the expression products showed obvious cytotoxicity, just in agreement with the prediction of modeling.]]> PEI Wu-Hong,FENG Jian-Nan,LEI Hong-Xing,LI Yan and SHEN Bei-Fen 1829true －6～1.91×10－3 mol/L for triglyceride with a slope of 32.7 mV/pC in the pH 8.5 Tris-HCl buffer solution at 37℃. The response time is 5～10 min. The sensor is relatively stable for 25 days.The sensor has been applied to the determination of triglyceride in serum with satisfactory results.]]> WANG Yong-Sheng,LI Gui-Rong,Lü Chang-Yin,WAN Zai-Yang and LIU Chuan-Xiang 1828true Dunaliella salina under the special stress conditions.The results show:1.The strength of stress conditions is advantageous to accumulate trans-isomer, otherwise,decreasing the stress conditions advantage to accumulat cis-isomers. For example, under the conditions of 32℃ with light intensity 10 000 lx, salinity 18%, the amount of trans-isomeris most (2.593 mg/L), but the cis-isomers amount is most (0.630 mg/L) under 25℃, indoornature light and salinity 24%.2.The kind and amount of cis-ismoers are different when the stress conditions are changed.After being cultured 30 days,trans-isomer and two cis-isomers were obtained under 25℃,indoor nature light salinity 24%,but only trans-isomer and one cis-isomer were obtained,when 32℃ with 10 000 lx light intensity and salinity 18%.3.Eventhongh the stress conditions is the same, it is different of the isomers'kind and amount only when culturing time is changed.For example: under the conditions of 25℃ 10 000 lx,18% salinity, only trans-isomer was found before culturing,after 8 days and 19 days,trans-isomer and two cis-isomer, after 34 days,trans-isomer and one cis-isomer were found.]]> WANG Yong and QIAN Kai-Xian 1827true ZHONG Shu-Ping 1826true WANG Jian-Zhi,GONG Cheng-Xin,I.GRUNDKE-IQBAL and K.IQBAL 1825true + in dopaminergic neuronal cell line (MN9D) was investigated with repatched whole-cell method. It was found that the voltage dependent calcium current (ICa) was significantly suppressed in differentiated cells and ICa induced by high depolarizing pulse decreased earlier than that induced by low depolarizing pulse during MPP+ treating for up to 1 h. In undifferentiated cells, no apparent change happened under the same condition.]]> CAI Dong,WANG Xiao-Min,QU An-Lian,HAN Song-Ping,YU Ying-Xin,ZHOU Zhuan,KANG Hua-Guang and HAN Ji-Sheng 1824true -·2) from the respiratory burst while 10 g/L SPS inhibited the oxygen consumption of PMN markablely， and 1 g/L SPS scavenged 53.2% O-·2.]]> LI Yan,TIAN Xiao-Hua,CONG Jian-Bo,TENG Xia,SUN Cun-Pu and GU Jing-Fan 1823true 4 and transferred into, expressed in E.coli BD21(DE3) induced by IPTG. The expression level was up to 15% of total bacterial proteins. Mutated calponin was liberated form E.coli by freezing and thawing and purified by Sephadex G-50. The recombinant protein and the purified mutant protein was identified by Western blotting.]]> CAO Sheng,TANG Da-Chun and LUO Jian-Yun 1822true LIU Hong-Tao,WANG Yu-Gang,ZHANG Ying-Mei,SONG Quan-Sheng,JING Bao-Qian,YUAN Yong and MA Da-Long 1821true ΔAmax）of ACMA(9-anmino-6-chloro-2-meto-acridine) induced by ATP,NADH and succinate was significantly decreased (P＜0.05). The time of maximum fluorescent quenching（TΔAmax）and the time of half maximum fluorescent quenching（T1/2ΔAmax）were significantly prolonged (P＜0.01). It was suggested that the ability of proton translocation across the IMM in endotoxic shock rats was decreased. The membrane fluidity were significantly decreased at the deep level and hydrocarbon chain. The mitochondrial membrane-associated PLA2 activity, plasma MDA,and mitochondrial MDA contents were significantly increased in endotoxic shock rats. The results showed that PLA2, and lipid peroxide may be the causes of the changes of proton translocation in endotoxic shock rats.]]> YANG He-Ming,LU Song-Min,LIU Jian-Cang,LI Ping,ZHU Zheng-Kun and GUO Su-Qing 1820true HUANG Jian-Sheng,XIE Yong-Mei and REN Da-Ming 1819true Indocalamus tesselatus), a kind of bamboo leaves by alkaline extraction were studied. Their physicochemical properties and structural feature were elucidated by modern analytical technique such as IR, NMR, GC, partial acid hydrolysis, periodate oxidation and Smith degradation. The typical absorptions of these polysaccharides were shown in IR spectra. The main structure was examined: FⅢ-a, (1→6)-β-D-galactosyl branched (1→3)-α-D-Xylan and FⅣ-a, (1→6)-β-D-galactosyl (1→3)-α-D-Xylan with arabinose branches at C-6 position.]]> CHEN Chun-Ying,DING Yu-Qiang,E.A.ELMAHADI,ZHOU Jing-Yan,LI Yan and XU Hui-Bi 1818true QIU Guang-Liang,Deligeer,LI Shu-Yuan and GAO Yan-Chun 1817true NI Ju-Hua,LIU Yun-Hao and JIA Hong-Ti 1816true 2+ in A549 cells sensitive and A549/DDP cells resistant to the cis-dichlorodiammine platinum (cisplatin) were measured by Fura-2/AM, the proliferation ability and cell cycle were measured by propidium iodide(PI) labeling cellular nuclear DNA. The results indicated that the concentration of intracellular free calcium of the sensitive A549 cells was 2 times higher than that of the resistant A549/DDP cells; the proliferation ability of the latter increased significantly than that of the former, the cell cycle also shortened. The proliferation ability and cell cycle of the two cell lines also clearly showed difference by decreasing or increasing their intracellular free calcium concentration if the cells were treated with BAPTA-AM or EGTA and A23187 or Thapsigargin. All of the results demonstrated that the concentration decrease of intracellular free calcium in the A549/DDP cells resistant to cisplatin may affect the cellular proliferation, shorten the cellular cycle, which would be helpful to remain the multidrug resistance characteristics of A549/DDP cells by specially modulating the cellular decisive G1 of cell cycle.]]> LIANG Xing-Jie,HUANG Zhen-Hua,LU Yan-Meng,LI Yu-Mei and HUANG You-Guo 1815true In vivo, it can transfer the light energy into photosynthetic center, however, it can deliver the captured light energy captured to the surrounding oxygen in vitro and produce reactive oxygen species such as singlet oxygen, which is toxic to tumor cells. R-PE was added to the culture medium of tumor cells, subsequently with irradiation of 488 nm, Argon laser of 25.6 J/cm2. The result by MTT assay showed that the survival rate decreased with the increase of R-PE concentration from 1 to 100 mg/L. The result from 3H-TdR incorporation demonstrated that the synthesis of DNA reduced when the concentration of R-PE increased from 0.01 to 0.32 mg/L. Besides, pUC18 DNA showed a conversion from supercoiled into linear conformation. The conclusion comes that R-PE mediated PDT can influence the conformation of DNA, and it may be one of the mechanisms of R-PE mediated photodynamic therapy .]]> LI Guan-Wu,WANG Guang-Ce,LI Zhen-Gang and ZENG Cheng-Kui 1814true +-ATPase is one of the ATPases and play an important role in the growing period of the plant. Hypocrellin B and KI quench the intrinsic fluorescence of outside and inside membrane domain respectively. This two quench probes have been used to quench the protein’s intrinsic fluorescence under different pH and temperature. The relationship between hydrolysis activity and folding condition of V-ATPase has been preliminarily studied. The KSV of outside and inside membrane domain under different pH and temperature had been compared. It shows that the intrinsic fluorescence of the protein and KSV of outside and inside membrane domain all dropped with the deviation of pH and temperature from the optimum condition and the activity of the enzyme dropped too. This illustrates that the folding condition had been changed with the dropping of the enzyme’s activity. The changing of the folding condition of the protein plays an important role in the inactivation mechanism.]]> DONG Cai-Hua,WANG Zhi-Qiang and WANG Yan-Zhi 1813true 2O2 over a wide concentration range (0.05～50 mmol/L) to assess dynamically the effect of H2O2 on cardiomyocytes. The results showed that application of ＜0.1 mmol/L H2O2 to cardiomyocytes caused accumulation of lipid peroxide(MDA) at 24 h, and phase changes of cellular proliferation cycle at 2h, which represented early biochemical changes. Exposure of cardiomyocytes to the increasing concentrations of H2O2 (1～5 mmol/L) induced progressively biochemical injury; the levels of LDH and MDA were significantly higher in the cardiomyocytes exposed to H2O2 than in control, with concomitant morphologic changes. When exposed to high concentrations (＞10 mmol/L), a large number of cardiomyocytes were found dead by MTT assay, and morphologic examination by HE-staining showed that cardiomyocytes contracted extremely, forming a large dropping areas of cardiomyocytes, concurrently with the marked increase of membrane permeability which caused LDH substantial leakage from myocytes to H2O2. It is proposed that H2O2 accumulation can induce cardiomyocyte cytotoxicity in a dose-and time-dependent manner. Treatment with low concentrations of H2O2 causes cardiomyocytes early slight biochemical changes which represent pre-apototic injurious features. High concentrations of H2O2 can progressively induce lipid peroxidation, which cause the severe damage of the cell membrane. With exposure of cardiomyocytes to H2O2, the magnitude of the cytotoxicity is modulated by horseradish peroxidase (HRP). It is suggested that HRP may protect cardiomyocytes against reactive oxygen species.]]> CAO Chun-Zhang,BU Li-Sha,GAO Shen and Yang Tong-Shu 1812true E.coli, and the product of expression was purified.]]> CHENG Xi-Ping,XIAO Hua-Sheng,HUANG Wen-Jin,ZHANG Ping,JU Gong and ZHANG Xu 1811true Populus euphratica(PE) cells had been isolated and purified by two-phase partition composed of Dextran T-500 and PEG 3350. Effect of different concentrations of the polymer (5.5%，5.7%，5.9%，6.1%，6.3%，6.5%) and KCl (0，5，10，15 mmol/L) on the purification were examined. The results indicated that two-phase system composed of 5.9% polymer without salt resulted in plasma membrane of PE with higher H＋-ATPase activity. The activity of the membrane H＋-ATPase has been increased 8-fold (from 4.23 to 32.63 μmol/mg·h) with better orientation and most of the membrane H＋-ATPases are right side-out. These results may provide better basis and availability for further studies of the purified H＋-ATPase from PE plasma membrane.]]> ZHANG Ling,LIU Qun-Lu and HUANG You-Guo 1810true Candida rugosa lipase (CRL) is fractionated into four fractions containing three isoenzymes (CRL-1, CRL-2 and CRL-3). They have different enantioselectivity for the asymmetric hydrolysis of (R, S)-Naproxen methyl ester in the aqueous-organic solvent biphase system. As analyzed on SDS -polyacrylamide gel electrophoresis, isoelectric focusing and organic solvent treatment, slight structural difference among these isoenzymes has been found. CRL-1 and CRL-2 are associated noncovalently with low molecular mass acidic components in different degree, CRL-3 is disassociated with the low molecular mass acidic components. Because of the perhaps hamper of the low molecular mass acidic components, the open extent of hydrophobic pocket around active site on CRL-3 is bigger than on CRL-1 and CRL-2. According to the slight structural difference, isoenzymes with different enantioselectivity for hydrolysis of Naproxen ester have been immobilized selectively on different hydrophobic supports (GDX101 and YWG-NH2). Via a sample and easily performed selectivity adsorption step, the purification can be combined with the immobilization. This new method seems to be very suitable for an easily separation of such isoenzymes with slightly different structure.]]> XIN Jia-Ying,XU Yi,LI Shu-Ben and WANG Lai-Lai 1809true Agkistrodon acutus forms a 1∶1 complex with activated coagulation factor Ⅹ (FⅩa) in the presence of Ca2+ ions. The result that ACFⅡ fails to form a complex with FⅩa in the absence of Ca2+ ions indicates that the binding is in the Ca2+-dependent manner. ACFⅡ is a new member of the Ⅸ/Ⅹ-bp family, and has a similar amino acid composition to the other members of this family. It is composed of 251 amino acid residues with a relative molecular mass of 29 468.1, determined by MALDI-TOF-MS. ACFⅡ prolongs plasma prothrombin time (PPT) in the dose-dependent fashion and exhibits marked anticoagulant activity only at the concentration higher than the critical concentration (12 nmol/L) of ACFⅡ.]]> XU Xiao-Long,LIU Qing-Liang and WU Shuang-Ding 1808true 4)2SO4 precipitation. Protein gel was generated by addition of thrombin to the complex composed of plasma protein, bovine fetal serum and DMEM.Bovine aortic endothelial cell(BAEC) was plated on the gel matrix and the growth，vascular formation of endothelial cell in the gel were observed. The results demonstrated that BAEC can adhere to the gel surface and grow normally as grow on the culture plate, while under the induction of bFGF, BAEC can migrate into the gel and form vascular-like structure in the gel matrix. Many vascular-like structures can link to each other to form capilary net structure.]]> LI Fu-Yang,YAO Li-Bo and YANG Jing-Hua 1807true YANG Jian-Hua and QIAN Xia 1806true 5 CFU/ml) retrovirus supernatant, and by selection with G418 positive cell clones (T/pCD2) were obtained. PCR and RT-PCR showed that CD gene was transferred into the T-lymphocyte and expressed successfully. T-lymphocyte and T/pCD2 cells were exposed to different doses of 5-FCyt for various hours，and were then observed under light microscope and the viability of these cells was determined by MTT colorimetric cell proliferation assay. The results showed that retrovirus-mediated CD gene transferred to T/ pCD2 cells could confer them high sensitivity to 5-FCyt, and T/ pCD2 cells exposed to 5-FCyt (＞1 μmol/L) can be killed. But 5-FCyt was not harmful to normal T-lymphocyte cells. The survival time of T/ pCD2 cells in the presence of 5-FCyt(3～5 d) was significantly shorter than that of T-lymphocyte(＞14 d).]]> SONG Yan-Bin,WU Zhi-Jian,YIN Fang,MA Wen-Li,YANG Guang-Cai,WU Bo-Song and XU Qian 1805true 32P dCTP in the presence of 2′, 3′-dideoxy-cytidine-5′-triphosphate (ddCTP) by terminal deoxynucleotidyl transferase (TdT) was developed. The saturation labeling of 3′-ends of DNA fragments was performed by adding different concentrations of α-32P dCTP to a DNA sample, from which a maximal labeling (Lmax) and a kinetic parameter(Km) of the TdT reaction were calculated. Results were confirmed by agarose gel electrophoresis, fluorescein-dUTP and exogenous teminal deoxynucleotidyl transferase(TUNEL), flow cytometric analysis (FCA). The method mentioned above requires as little as 5 ng of DNA, increases in the sensitivity of DNA fragments detection by at least 200-fold relative to the widely used agarose gel electrophoresis, and the linearity of the assay is about 5～5 000 ng DNA. The application of the method in the apoptosis study showed that(1) a time- and dose-dependent increase in the number of DNA strand breaks in apoptotic Raji lymphoma lymphocytes induced by dexamethasone, and (2) age-dependent increase in the number of DNA strand breaks occurred in the cardiac tissues of spontaneously hypertensive rats (SHR) compared with that of normal control rats (WKY). Results of the assay were confirmed by the DNA ladder pattern exhibited after electrophoresis, fluorescein-dUTP and exogenous terminal deoxynucleotidyl transferase(TUNEL), flow cytometric analysis(FCA)(r＞0.98). It is a quantitative, simple, sensitive, specific and useful assay for assessing DNA degradation in molecular and cell biology especially in apoptosis research.]]> PENG Li-Ming,JIANG Hong,J.J.LIU and C.J.BRADLEY 1804true 8) on calmodulin activity in rat cerebral cortex，the cerebral cortex neurocytes was used as a model. CCK8 stimulated the activation of CaM in a time-dependent manner. After 15 minutes of treatment of 1 μmol/L CCK8，the CaM activity reached the maximum increase. CaM activity was increased in a dose-dependent manner by CCK8 (10－12～10－7 mol/L). The CCKB-specific receptor antagonist L-365，260 and with a weaker efficiency，the CCKA-specific receptor antagonist L-364，718，were able to block a maximal effect of CCK8-induced CaM activation，suggesting that CCKB receptor may regulate CaM activity in cerebral cortex.]]> XIANG Peng,CHEN Man-Ling and WU Zhao-Feng 1803true HUANG Xing-Xu,CHEN Lian-Bo,BAO Yong-Yao,PIAO Ying-Jie and HUANG You-Guo 1802true CUI Da-Xiang,YAN Xiao-Jun,WANG Feng,ZHAO Jin-Rong and SU Cheng-Zhi 1801true ZHANG Yan,SHI Xiu-Fan and LIU Ci-Quan 1800true in-situ in a fermentation process. The presence of microbial cells would affect the dielectrical properties of microbial suspensions. At radio-frequencies the dielectric permittivity of cell suspensions are monotonic function of the measuring frequency and cell density. Based on this understanding a new method for measuring biomass concentration is proposed. Using this method the biomass concentrations are measured on-line and in-situ without taking samples from the bioreactor, and only viable cells are detected. The electrode could be directly inserted into the fermentor and could be sterilized in place. The method has a wide use in the industrial field of pharmacy, brewery, sewage disposal.]]> WANG Yi-Jun,FAN Yu,L.OLSSON and J.NIELSEN 1799true Escherichia coli. The results showed that such protein was successfully prepared and its bispecific activities were testified by affinity blot and Western blot. The biotin binding and immunocytochemistry assay indicated that the expressed fusion protein was capable of binding biotin molecule and CEA antigen respectively. This protein has the potential application when combined with the corresponding tests.]]> REN Jun,JI Wan-Sheng,ZHOU Shao-Juan,CHEN Zheng,ZHANG Xue-Yong and FAN Dai-Ming 1798true MIF) of the three different hepatocytes were determined and calculated after simultaneously incubated with FITC-NGA at the same concentration. The concentration of FITC-NGA for saturating ASGPR on the surface of L-02 was 0.4 mg/L, at which the MIF of the three different hepatocytes were 228.7，5.81 and 1.13 respectively. The saturated combination can be completely inhibited by 50-fold NGA or 10mmol/L EDTA. FCM can display soundly the characteristic of the receptor-ligand combination between ASGPR and FITC-NGA. Compared with normal rat hepatocytes, there is no ASGPR on the surface of BEL-7402, and the quantities of ASGPR on the surface of injured rat hepatocytes decreases significantly.]]> LI Wen-Xin,ZHANG Rong-Jun,TAN Cheng,TAO Yong-Hui and JIN Jian 1797true Ki) is 3.86 mmol/L.The activity inhibition of PLAP also affected by the pH, its activity inhibition disappeared below pH 7.5,but gradually increasing between pH 7.5～10.5. The results suggest that the activity inhibition and the conformation changes of the enzyme were caused by PCP. The inhibition of PLAP activity correlates with the dissociation state of PCP.]]> GENG Fang-Song,WANG Xiu-Li,TONG Jia-Ming and LIU Hong-Ming 1796true 6-tag) at the N-terminus. Low bleeding of metal ion was achieved. It was proved that the properties of IMAC membranes were better than conventional chelating sepharose fast flow column. The fractionation of recombinant proteins that carry a polyhistidine tag is currently perhaps the most promising application of IMAC.]]> WEI Qi,YAO Ru-Hua and BAO Shi-Xiang 1795true BAI Hua,LU Jing-Fen,YAN Zhen-Ying,GULI Nuer and ZHANG Min 1794true Anthopleura qingdaoensis contains at least two cardiotonic polypeptides capable of enhancing cardiac contraction(a positive inotropic effect). An artificial gene encoding anthopleurin-QD2 (Ap-QD2), which consists of 49 anino acid residues, was cloned and transferred into pPICZαA, a secretory expression vector for Pichia pastoris. The construct was linearized and was integrated into the yeast chromosome by electroporation under the selection of zeocin. Approximately 20 mg/L of biologically active Ap-QD2 was produced from one of the KM71(Muts) transformants, and about 7 mg of pure Ap-QD2 was obtained after chromatography purification.]]> WANG Wei-Rong,LIU Ai-Min,QIAN Zhi-Kang and HUANG Wei-Da 1793true Pleurotus tuber-regium （Fr.）Sing. was prepared by the reaction of HNP with monochloroacetic acid. The antioxidative activities of CM-HNP were also studied. The results showed that CM-HNP can protect liver mitochondria from lipid peroxidation induced by Fe2+-Vit C including the increase of TBARS contents, the swelling of mitochondria and the decrease of membrane fluidity with a dosage-effect manner. In addition, CM-HNP can effectively scavenge O-·2 generated by the self-oxidation of pyrogallic acid.]]> WANG Yan,YANG Xiang-Liang,DENG Cheng-Hua,GU Xiao-Man,ZHOU Jing-Yan and XU Hui-Bi 1792true in vitro mutagenesis, then the interactions of IGF-1 and various mutants IGF-R1 were quantified to identify the gain-of-function mutation in the yeast two-hybrid system by assaying the β-galactosidase activity. The results confirmed the binding data of IGF-1 to IGF-R1 and suggested that the residues N237 and T238 on IGF-R1 play a crucial role in binding to IGF-1.]]> DU Qing-You,LIU Bao-Ying,XUE Yan-Ning,WANG Hui-Xin and DING Hong-Mei 1791true FENG Jian-Nan,REN Yun-Fang,NI Chang-Yuan,WU Jia-Jin and SHEN Bei-Fen 1790true 2 (A.aBPLA2) from Agkistrodon acutus was inserted into a bacterial expression vector pBLMVL2 and effectively expressed in E.coli RR1. The protein was produced as insoluble inclusion bodies. After partial purification by washing the inclusion bodies with Triton X-100, denaturing and refolding, the renatured recombinant protein was purified by FPLC column superoseTM12. The enzymatic activity of the expressed A.aBPLA2 is close to those of denatured-refolded native acidic phospholipase A2 from Agkistrodon halys Pallas, A.aBPLA2 has the same hemolytic activity as denatured-refolded basic phospholipase A2 from Agkistrodon halys Pallas, but its inhibiting effect on platelet aggregation is negligible. The roles of various amino acid residues in the enzymatic activity and pharmacological activities of phospholipase A2 are discussed.]]> LIU Xiao-Long,ZHONG Xiao-Yan,WU Xiang-Fu and ZHOU Yuan-Cong 1789true Penicillium was immobilized on chitosan by glutaraldehyde.The results indicated that the immobilized-hemicellulase prepared by 0.5 g chitosan crosslinking with 4% glutaralhyde and then combining with 2.5 mg protein showed higher enzyme activity and better activity recoveries(45.6%).The optimum pH of soluble enzyme and immobilized enzyme were pH 4.6 and pH 3.6 respectively.The optimum temperature of soluble enzyme was 55℃, whereas immobilized showed high activity in 60～75℃.Thermal stability of immobilized enzyme was better than soluble enzyme at 65℃.The apparent Km′ of the immobilized enzyme was 3.58×10－2 g/L and the Km of soluble enzyme was 5.0×10－2 g/L with hemicellulose as the substrate.]]> ZHU Qi-Zhong 1788true ZHANG Xiao-Bo,XU Xun,LI Qing-Ge and XU Li-Mei 1787true 3O4 particles and soybean licithin which was only 20 nm in diameter. The heat effect was studied under 10 kW, 100 kHz magnetic field. The temperature change was measured by a glass thermometer (±0.1℃) in agarose gel and distilled water with different Fe3O4 content respectively. The effect of magnetic intensity was discussed. This was an experimental basis for hyperthermia treatment tumors. The results indicated that the tempereature raised quickly as the increase of Fe3O4 concentrations and the magnetic field intensity. The tempereature finally remained constant due to the balance of heat generation and heat transfer to the surroundings. When Fe3O4 concentration were 1 g/L and 2 g/L, the balance temperature were respectively 41.9℃ and 47.5℃ as the magnetic field was 2.78×104 A/m. In 1 g/L Fe3O4 concentration, the magnetic field intensity were 2.78×104 A/m and 1.11×104 A/m, the temperature plateaus were at 41.9℃ and 30.7℃ respectively. The heat transfer faster in water than in agarose gel.]]> SHI Feng and WU Min 1786true YANG Jie and LIU Ci-Quan 1785true Agkistrodon halys has been used to investigate its effects on MAPK and MAPKK of PC12 cells. The results showed that this kind of NGF increased MAPK and MAPKK expression and MAPK activity. The above increases were NGF concentration dependent in the range of 25～100 μg/L. The use of PKC inhibitor H-7 indicated that the increased expressions of MAPK and MAPKK expression were PKC-dependent.]]> QIN Li-Ya,LIU Yu-Xin,ZHANG Lei,SONG Ling-Li and CUI Zhao-Chun 1784true JIANG Guo-Chun,YUAN Li-Zhen,GUO Xue-Min,WEI Kang,ZHANG Xue-Feng and JIA Xiang-Xu 1783true SUN Run-Guang,ZHANG Jing and WANG Yong-Chang 1782true + RNA were isolated from cell lines of RCC and normal kidney respectively.Moreover, single-strand cDNAs and double-strand cDNAs were synthesized in turn. After enzyme restriction,cDNAs between 400～600 bp were obtained. RCC cDNAs then were divided into two groups and ligated to the specific adaptor l and adaptor 2 respectively .After RCC cDNAs hybridized with normal kidney cDNA twice and underwent two times of nested PCR,then with arms of T/A plasmid vectors to set up the subtractive library. Amplification of the library was carried out with the E.coli strain Top 10F′. Human RCC subtrctive library with high subtractive efficiency was set up sucessfully. The amplified library contains 6 500 positive clones.Random analysis of 350 clones with enzyme restriction shows that all plasmids in the clones contain 400～600 bp inserts. The constructed cDNA subtractive library of human RCC is a highly efficient one and lays solid foundation for screening and cloning new and specific oncogenes or tumor suppressor genes of RCC.]]> ZHANG Qiang,MAO Ze-Bin,ZHANG Zhi-Wen,XIN Dian-Qi and GUO Ying-Lu 1781true HAN Wen-Ling,LI Ying,YANG Gao-Yun,SONG Quan-Sheng,ZHANG Ying-Mei,DI Chun-Hui and MA Da-Long 1780true Q=1.8382－2.5186×10－2T(r=0.9913).Based upon the various assays used to measure the entrapment efficiency and diameter of pH-sensitive liposomes, it was concluded that the entrapment of antisense oligodeoxynucleotides by pH-sensitive preliposomes was effective.]]> WANG Hong,WANG Sheng-Qi,WANG Zhi-Qing and ZHU Bao-Zhen 1779true WAF1 gene expression decreased obviously and gradually,bcl-2 gene expression did not decrease until apoptosis happened, c-myc and c-fos genes expression enhanced, while p53 and HER-2 genes expression had no obvious change. To analyze the change of p21WAF1 promoter activity, 2BS-WP cells were used, which were stabely transfected with pDOR-WPs that containing different length of p21WAF1 promoter followed by green fluorescence protein (GFP) report gene. The results showed that the level of GFP expression decreased during NaBu induction, and the main regulating region was 0～－800 bp at upstream of TATA box of p21WAF1 promoter. It is suggested that down-regulation of p21WAF1 promoter activity was involved in the process of NaBu-induced apoptosis in 2BS cells, which was possibly p53-independent.]]> HUANG Ying,ZHANG Zong-Yu and TONG Tan-Jun 1778true LEI Hai-Xin,YU Lei,SUN Li-Jun and CHEN Chu-Chu 1777true LI Bin,LIU De-Pei,WANG Jing,DONG Wen-Ji,GUO Zhi-Chen,XIA Wei and LIANG Chich-Chuan 1776true Bacillus sp.2 by combined consecutive treatment consisting of ammonium sulfate fractionation, Q Sepharose FF, Sephadex G-100(Ⅰ), Hydroxyapatite and Sephadex G-100(Ⅱ) column chromatographies. The specific activity of purified 3-DG metabolizing enzyme was 63.75 U/mg. The molecular weight of the enzyme was about 32 ku. 2-Oxoaldehyde compounds were found to be specifically good substrate for this reductase. The optimum pH of the enzyme activity was 6.2. The enzyme was stable in the pH range from 5 to 8 and in the temperature range from 25℃ to 30℃. The Km for 3-DG was 2.3 mmol/L. Suitable amount of EDTA、β-mercaptoethanol and dithiothreitol enhanced the enzyme activity, but the activity of the enzyme was partially lost by adding iodoacetic acid or N-ethylmaleimide.]]> LIANG Zhi-Qun,SU Gui-Jiao,LI Xiang-Ping,MO Bo-Li and LIANG Jing-Juan 1775true YANG Xiang-Dong,YANG Yong-Zong and LI Jian 1774true FENG Jian-Nan,REN Yun-Fang,LI Song and SHEN Bei-Fen 1773true Saponaria officinalis.The molecular weights of SO3a and SO3b are about 22 500,19 400,respectively,and their pI values are about 8.4 and the amino acid components are analysed.The contents of the secondary structure of SO3a and SO3b are determined by laser Raman spectrum.In the experiment it was also found by reverse phase capillary chromatography that SO6 which was reported by stripe to be a purified protein,contain two components in fact.]]> YANG Yuan-De,ZHOU Kang-Jing,PAN Ke-Zhen,ZHANG Rong-Zhen,CHEN Ru-Ming and RAO Ping-Fan 1772true 5、β1 subunits. Further study on the content of integrin α5、β1 subunits——the major receptor of Fn on the cells’ surface showed that after the TPA acts on cell for 24 hours, the content of integrin α5、β1 subunits increased 52.3% and 51.6% respectively. To analyze the total content of N-oligosaccharides and its constituent propertion after TPA acts on cell, 3H-mannose incorporation into N-oligosaccharides and lectins chromatography were used. The result has no difference from that of control. It can be said that increasing adhesion is induced increasing the syntheses the content of α5 and β1 subunits of the cells. The PKC inhibitor Sphingosine was added during TPA acting on cell, it was found that both the α5 and β1 content and the adhesion of cells to Fn recovered to the control level. It proved that increasing α5β1 synthesis thus increasing adhesion of cell to Fn by TPA is mediated by PKC. Besides the inhibitor of tyrosine protein kinase also can block the action of TPA increasing the content of α5β1 integrin syntheses.]]> HE Jian-Yu,FANG Xin-Chu,CAO Li-Huan and ZHA Xi-Liang 1771true Halobacteria’s cell structure. It a simple but ingeniously contracted photoelectric transducer. The process of its synthesis is greatly affected by the compositions of culture medium and the culturing time. The results showed that optimum culture medium of the purple membrane synthesis is synthetic medium and optimum-culturing time is 7 days.]]> DILBAR Tohty,HUI Shou-Nian,XU Xiao-Jing and ZHOU Pei-Jin 1770true Pseudomonas pseudoalcaligenes insecticidal protein was isolated and purified sucessfully. Amino acid composition analysis showed that it is an acid protein. Laser Raman spectra of the insecticidal protein have been obtained and analysed.It is found that protein of the sample contains predominantly β-sheet content.The majority of tyrosyl residues are exposed and a few are buried in the protein of the sample.The sample has trans-gauche-trans configuration of C—C—S—S—C—C linkage.]]> LUO Xia,YANG　Ｚｈｉ-Rｏｎｇ,ZHU　Ｗｅｎ and ＬIU　Ｓｈｉ-Gｕｉ 1769true YU Min-Zhong 1768true XU Li-Yan,LI En-Min,XIONG Hua-Qi,CAI Wei-Jia and SHEN Zhong-Ying 1767true YAO Bao-Li,LI Bao-Fang,LI Xing-Chang and N.HAMPP 1766true HE Yun-Gang,TAN De-Yong,QIAN Wei,LAI Jian-Hua and XIE Yong-Fang 1765true Selenocosmia humena, which has analgesic activity. According to the RT-PCR result, an artificial gene encoding HWAP-Ⅰwas chemically synthesized. The gene was then inserted into pPIC9K,a secretory expression vector for Pichia pastoris.The constructed pPIC9K-HWAP-Ⅰ was transformed into his4 mutant yeast GS115 and a Muts His+ cell line was screened.The recombinant HWAP-Ⅰwas secreted into the culture supernatant induced by 0.5% methanol.Tricine SDS-PAGE analysis indicated efficient expression and secretion of HWAP-Ⅰ.The molecular mass of product was about 4 ku. Yield of product was estimated to be 80 mg/L. Purified protein was assayed for biological activity and the results demonstrated that HWAP-Ⅰ had 60% analgesic activity compared to native HWAP-Ⅰ.]]> HE Ning-Jia,NIE Dong-Song,XU Hui-Ming,LI Min and Liang Song-Ping 1764true Codium fragile, by Triton X-100 sucrose gradient centrifugation. Zone Ⅲ had a Chl a/b＞20, and designated as PSⅠ core complex CCⅠ because it created only CPⅠ band in mild PAGE. Zone Ⅳ and Ⅴ had absorption at 436 and 674 nm, 467 and 650 nm, and 540 nm, suggesting the presence of Chl a, Chl b, siphonaxanthin and siphonein, Chl a/b were 3.23 and 2.4, respectively. Both CPⅠ and CPⅠa bands were observed when they were subjected to mild PAGE. Therefore, Zone Ⅳ and Ⅴ were different forms of PSⅠ complexes that consisted of CCⅠ and different amount of light-harvesting complex LHCⅠ. Zone Ⅲ contained only 66 and 56 ku peptides in SDS-PAGE, while Zone Ⅳ and Ⅴ had 4 different LHCⅠ peptides of 25，26，26.2 and 27.5 ku in addition to 66, 56 ku peptides. Fluorescence emission spectra showed that efficient energy transfer were kept among pigments in isolated PSⅠ complexes. Excitation energy absorbed by Chl b, siphonaxanthin and siphonein can be transferred to Chl a.]]> CHEN Min,LI Ai-Fen and ZHOU Bai-Cheng 1763true E.coli. After denaturation and renaturation, the aim protein was purified to homogeneity by Zn2+ chelating chromatography and Sephacryl S200 chromatography. The apparent molecular mass was 35 ku shown by SDS-PAGE analysis. The special activity was 87 000 U/mg detected by fibrin plate determination. The enzyme had similar kinetic parameters to that of natural uPA-32k when was assayed with the chromognic substrate S2444. However Fβ(15～42)/scuPA-32k had higher fibrin affinity than that of natural scuPA-32k and had antifibrin polymerization. These results showed that the fusion protein had good respects.]]> JIAO Jian-Wei,LIU Ning,YU Mei-Min and RU Bing-Gen 1762true Ctenopharyngodon idellus）cell culture ZC7901 was used as an in vitro model system to study the effect of heavy metal ions on fish cells. The results showed that the cell culture chemically stressed by six heavy metal ions, Cd2+，Cr6+，Hg2+，Cu2+，As5+ and Pb2+, exhibited cell death accompanied by various morphological alterations such as nuclear condensation, chromatin abutted sharply against nuclear membrane and developed into distinct apoptotic-like bodies, which were characteristic features seen in apoptosis. By agarose gel electrophoresis, DNA ladder were clearly visualized, which indicated the specific degradation of DNA. Also terminal-deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL) analysis of the cells demonstrated DNA fragmentation localized in the dying cells as well as in the additional apoptotic-like bodies. All the results indicated that six heavy metal ions, Cd2+，Cr6+，Hg2+，Cu2+，As5+ and Pb2+, were all able to induce apoptosis in fish cells.]]> XIANG Li-Xin,SHAO Jian-Zhong and MENG Zhen 1761true 2+ and ionophore A23187 on erythrocyte deformation, the new ektacytometry is used. After erythrocytes were treated with different concentrations of intercellular Ca2+ and ionophore A23187 respectively, the orientation index (DI)or and small deformation index (DI)d of the erythrocytes were measured. It is suggested that the effect of ionophore A23187 on the rheology of erythrocytes is more notable than that of intercellular Ca2+. Furthermore, the (DI)or,max and (DI)d,max decrease as the concentration of intercellular Ca2+ or ionophore A23187 is increased. The deformability of erythrocytes was obviously reduced after treated with ionophore A23187.]]> WANG Chun-Yi,ZENG Yan-Jun,WEN Zong-Yao,KA Wei-Bo,YAO Wei-Juan and LIAO Dong-Hua 1760true 2 for 2 weeks were autopsied, the brain proteins were extracted and separated by gel filtration, and further analyzed by 2-D PAGE, More than one hundred water-soluble proteins of rabbit brain were visualized by Coomassie Brilliant Blue staining. The apparent molecular masses of most water-soluble proteins from rabbit brain were about 6～30 ku, pI was about 4～9. After matched with normal alive rabbit brain protein spots separated by 2D-PAGE, there obviously exists many differences displayed and unmatched protein spots in the 2-D PAGE maps of Pb2+ induced rabbit brain. 3 spots were extracted from the 2D-PAGE and detected by the MALDI-TOF-MS and ESI-MS, The experiments show that rabbit brain proteins would be affected by Pb2+ injection.]]> ZHAO Rui,JI Jian-Guo,HAO Shou-Jin,CHEN Qian,YUAN Hong-Sheng,GUAN Hua-Shi and RU Bing-Gen 1759true in vitro growth inhibitory activity on cultured neuron; but the underlying molecular mechanism remains unknown. To probe this mystery, the yeast two-hybrid system was employed to screen a human brain cDNA library for GIF-interacting proteins. From 4.1×106 transformants of library screening, a C-terminal fragment of small G-protein Rab3a was obtained, harboring 87 amino acids that interacted with GIF in yeast. The full-length Rab3a was amplified by polymerase chain reaction (PCR) using human total placenta cDNA as template and concomitantly cloned. In yeast two-hybrid test, full-length Rab3a also interacted with GIF. Through coimmunoprecipitation and Western blotting experiments, interaction between GIF and Rab3a was confirmed in mammalian cell. Furthermore, the result indicated that Rab3a interacted with GIF in a GTP-binding form.]]> KANG Qiao-Hua,CHEN Qiao-Lin,REN Hong-Wei and RU Bing-Gen 1758true 1FO-ATPase from pig heart mitochondria are investigated. It is found that Pi not only takes an active part in ATP synthesis as a substrate, but also has an inhibition effect on ATP synthesis of F1FO-ATPase. In the presence of 1 mmol/L ADP the inhibition increases with the increase of Pi concentration from 0.01 mmol/L to 10 mmol/L. The viewpoint is different from that azide only inhibites ATP hydrolysis but not affects the ATP synthesis at low concentration (＜1 mmol/L). The results indicate that 0.1 mmol/L azide apparently activates ATP synthesis of F1FO-ATPase. The activation is negatively related to the concentration of Pi in the reaction system. When the concentration of Pi is kept at 0.1 mmol/L, with the increase of azide concentration, the apparent activation degree of ATP synthesis by azide is changed. When the concentrations of azide and Pi are equal, the apparent activation by azide is the biggest. The apparent inhibition of ATP synthesis by azide would not begin until the concentration of azide reaches near 0.5 mmol/L. After the concentration of azide is higher than 1 mmol/L, the uncoupling of F1FO-ATPase appears. The mechanism of the roles of azide and Pi is discussed.]]> MENG Xi,LI Sheng-Guang and LIN Zhi-Huan 1757true 2O2-induced mitochondrial PTP opening. GTPs and its major components EGCG, ECG have inhibitory effects on it, while other kinds of catechins, EGC, EC and (+)-C can accelerate the process. The data provide an alternate interpretation of the potent protective function of GTPs, EGCG and ECG.]]> SHEN Sheng-Rong,JIN Chao-Fang,CHEN Zi-Yuan,CAO Yuan-Lin and ZHAO Bao-Lu 1756true E.coli, resulted in the production of a protein has a subunit molecular mass of 71 ku and is recognized by both anti-Mn-SOD and anti-tetanus toxin antibody. The Mn-SOD moiety retains substantial enzymatic activity，where the TTC moiety can delivers the fusion protein to central nervous system neurons. Such fusions should provide a powerful tool for investigating the protective and destructive roles of Mn-SOD in motor neurons.]]> ZHANG Yan-Hong,HE Hua-Jun,YUAN Qin-Sheng,YANG Wei-Dong and WU Xiang-Fu 1755true Saccharomyces cerevisiae by glucose transport, a set of S.cerevisiae strains with variable expression of only one glucose transporter, Hxt7, the most abundantly expressed high-affinity transporter, was constructed. The strains were constructed by partial deletion of the HXT7 promoter in vitro and integration of the gene at various copy numbers into the genome of an hxtΔ (hxt1-hxt7 gal2 deletion) strain. The 149 bp DNA region －495 to －346 in the HXT7 promoter plays an important role in HXT7 expression. In the mutant strains with promoter length of more than 495 bp, the expression of HXT7 at high glucose concentrations was much higher than that in the wildtype strain. The level was dependent on copy number and promoter length. Increased expression at low glucose was maintained in these mutants. Hxt7 in the hxt null strain displayed an incomplete glucose repression. The growth rate correlated with the level of HXT7 expression at high glucose concentrations.]]> YE Ling,A.L.KRUCKEBERG,J.A.BERDEN and K.van DAM 1754true LI Fang-Ting and QI An-Shen 1753true gp37 genes of Spodoptera litura NPV (SpltMNPV) was cloned and sequenced. Employing computer analysis, glycosilated sites and the promoter motif existed in the gp37 gene. and was shown as a late gene encoding a 37 ku glycoprotein. A phylogeny tree was designed by the comparison of their homologues from known gp37 genes of baculoviruses. It was shown that the phylogeny of baculoviruses on the basis of gp37 gene was different from that based on polyhedrin genes. For example, Bombyx mori NPV (BmNPV) and Autographa californica NPV(AcMNPV) were evolved in the same branch, otherwise, they were put in two distinct branches in the tree according to the analysis of polyhedrin. This result was consistent with the baculovirus phylogeny based on EGT published previously.]]> LI Chong-Bi,LI Zhao-Fei,YAN Qing-Sheng,HU Guo-Dong and PANG Yi 1752true I, molecular mass and volume of protein in three different experiments were 4.43%±0.25%, 8.76%±5.14%, 13.00%±4.22% and 10.84%±9.16%, respectively. Totally, 996 and 1 256 protein spots were obtained in adult and aged murine brain map, repectively, of which 8 spots increased and 20 spots decreased in quantity. Furthermore, 4 spots were missing and 14 new spots emerged in aged murine brain compared with adult mice. The differentially displayed proteins between adult and aged murine brain are useful for diagnosing brain degenerative disorders.]]> DING Qin-Xue,QUE Hai-Ping,GUO Yao-Jun,JIN Shu-Qian and LIU Shao-Jun 1751true PAN Jun,WANG Yuan-Liang,CAO Xue-Bo,SU Lan,QIN Jian,LU Xiao and CAI Shao-Ji 1750true DING Li-Li,WENG Yi,ZHANG Yang-Rong and NI Da-Wei 1749true SUN Guo-Jing,TONG Xin,MENG Xiang-Bing,DONG Yan and SUN Zhi-Xian 1748true + memory T cells, mathematical models were set up respectively and the dynamics of different T subpopulations was studied. It was found that when invading antigen with optimal dose of, both models can generate memory and fit well with experimental data. Further study found that CD8+ T cell memory relates strongly to the persistence of antigen. Thus the contribution of antigen to maintenance of T cell memory is reconfirmed. The effect of the life-span of memory cells on immune memory was also investigated. Reverse linear differentiation model is deemed to have advantage in generation of immune response and memory.]]> MAO Li-Kai and QI An-Shen 1747true ZHAO Xiao-Rong,WANG Cheng-Xing,LUO Fei-Jun,GU Huan-Hua,TANG Min,XIA Lin-Qing,DENG Lin,YI Wei,DENG Xi-Yun and CAO Ya 1746true ZHANG Xiao-Hui,LI Zhong-Hua,ZHANG Bi-Cheng,DONG Li,ZHOU Ming,CAO Li,TANG Ke,LI Wei-Fang and LI Gui-Yuan 1745true Trichosanthes kirilowii, possesses anti-tumor and anti-human immunodeficiency virus (HIV) activities. With both two-photon laser scanning microscopy and confocal laser scanning microscopy in combination with the fluorescent probe Hoechst 33342, 2′, 7′-dichlorofluorescin diacetate (DCFH-DA), Indo-1 and Fluo 3-AM, TCS-induced changes in nuclear morphology, reactive oxygen species (ROS) and intracellular calcium concentration (［Ca2+］i) during the apoptosis of choriocarcinoma cells (JAR) were simultaneously observed for the first time. The results indicated that TCS-induced increase in ［Ca2+］i and ROS formation were involved in apoptosis of JAR cells, and that TCS-induced ROS formation was related to TCS-evoked increase in ［Ca2+］i. Further studies with confocal laser scanning microscopy revealed that TCS-evoked increase ［Ca2+］i was not the main factor responsible for TCS-induced ROS formation, and that TCS might induce the production of ROS through its interaction with membrane-bound receptor.]]> ZHANG Chun-Yang,GONG Yi-Xuan,MA Hui,AN Cheng-Cai and CHEN Die-Yan 1744true i of cisplatin-sensitive A549 cells decreased significantly, whereas the intracellular free Ca2+ concentration increased greatly with the culture time. But the mitochondrial membrane potential and pHi remained at a relatively high level and the intracellular free Ca2+ concentration was reduced gradually with the culture time for cisplatin-resistant A549/DDP cells. It would be suggested that the anti-apoptotic character of A549/DDP cells is related with the relative intracellular basification, the persistence of mitochondrial membrane potential and decrease in intracellular free Ca2+ concentration which would be responsible for the resistance of A549/DDP cells to cisplatin.]]> HUANG Zhen-Hua and HUANG You-Guo 1743true E.coli. The recombinant protein was purified. The renaturation occurred in the presence of sodium tetrathionate system. In order to study the biological activity of the recombinant protein, the effects of rhGDNF on the dorsal root ganglion (DRG) of chicken embryo (8 d), motor neurons of rat embryo (14 d) spinal cord and glial cell were explored. The results show that rhGDNF promotes the growth of neurite of DRG, increases the number of survival motor neurons cultured for 3 d, 7 d, 14 d and 21 d, respectively, and promotes glial cell proliferation.]]> WANG Jin-Hui,WANG Jia-Xi,DING Ai-Shi and LIU Hong 1742true in vitro by an assay of a single round of GTP hydrolysis on recombinant ARF1, and that GAP activity of ARFGAP3 was stimulated by PIP2 and inhibited by PC.]]> LIU Xiao-Qin,ZHANG Cheng-Gang,XING Gui-Chun,CHEN Qing-Tang and HE Fu-Chu 1741true Actinia cari. Toxins isolation was accomplished by freezing-thawing extraction, acetone fractional precipitation, cation exchange chromagraphy on SP Sephadex C 25 and reverse phase HPLC on ODS C18.They were found to be homogenous by SDS-PAGE and MALDI-TOF-MS, and their relative molecular masses are 5 008 and 4 992, respectively. They are found to be active on sodium channel in Guinea pig cardiac cell, RSAPI, increasing inward Na(+) currents, opens the sodium channel; RSAPⅡ, decreasing inward Na(+) currents, inhibits the opening of the sodium channel. The circular dichroic spectra of these two polypeptides are 100% β turn.]]> ZHUO Ru-Peng,FU Hong-Zheng,ZHONG Chong-Bo,WANG Xiao-Liang,LIN Wen-Han and ZHANG Li-He 1740true YANG Xue-Hui,LI En and TONG Xiao-Xu 1739true trans-factors, to certain extent, are dependent on the structural features of EE and PE in the secondary structure. According to the secondary structure of mRNA, the prediction accuracy of EE and PE sites may be improved.]]> ZHANG Jing,SHI Xiu-Fan and LIU Ci-Quan 1738true in vitro.]]> LU Cheng-Rong,LI Yong,DONG Chun-Na,LI Ming,WANG Qing-Ming,ZHANG Cheng-Gang,WEI Yu-Lin,CHEN Hui-Peng and HE Fu-Chu 1737true MA Shu-Hua,WANG Dun-Cheng,ZOU Zong-Liang,SHEN Bei-Fen and WANG Sheng-Qi 1736true Salmonella. A live attenuated AraA- autotrophic mutant of Salmonella typhimurium (SL3261) was used as carrier for eukaryotic expression vectors EGFPN1, pCMVmIL-12, pCMVmGM-CSF and was administered orally to BALB/c and C57BL/6 mice. After 6 weeks, these mice were challenged with 4T1 or Lewis tumor cells respectively. Flow cytometer and laser scanning confocal microscopy were used to detect the expression of GFP in murine tissues. PCR and ELISA were used to detect the integration and expression of mIL-12, mGM-CSF gene. The survival time of mice was also investigated. GFP expression and mIL-12, mGM-CSF gene integration could be detected in murine liver, spleen, intestine, kidney and tumor. The serum level of mIFN-γ，mIL-12 increased significantly in the mIL-12 orally treated mice (P＜0.05); The serum level of mGM-CSF increased also in mGM-CSF orally treated mice (P＜0.05); which resulted in the prolongation of the survival time of those mice, compared with the control (P＜0.05). Oral gene therapy using live attenuated Salmonella has the potential to be a simple, effective and above all, safe way against tumor.]]> LI Yu-Hua,CHEN Hui,GUO Kun-Yuan,XIE Yong-Mei,DU Jiang,ZHANG Ming-Hui,HUANG Jian-Sheng and REN Da-Ming 1735true LIU Sen-Lin,ZONG Min-Hua,TU Ran and ZHOU Fan 1734true 31P NMR) spectroscopy. The results led to a paradox of rapid intracellular pH recovery and a low level of high energy phosphate pools reserve during reperfusion compared to the control. It can be stated that Oxy improves energy consumption and, in turn, improves pH recovery, but also causes a deletion of high energy phosphate pools during reperfusion. The relationship between ions regulation, including H＋ regulation, and myocardial functions recovery may display a key factor for cell survival during postischemic reperfusion.]]> YAN Yong-Bin,LUO Xue-Chun,ZHANG Ri-Qing,WANG Xiao-Yin and FAN Li-Li 1733true CUI Da-Xiang,ZENG Gui-Ying,WANG Feng,TIAN Fu-Rong,GUO Yan-Hai,XU Jun-Rong,YAN Xiao-Jun,REN Dong-Qing and SU Cheng-Zhi 1732true LI Jun-Feng,WU Qi-Jiu,WANG Qian,JIANG Jin-Chang,JIN Hai-Qiang and LIN Yun-Fang 1731true E.coli JM109 after inducing by IPTG. The fusion proteins were purified by affinity chromatography on glutathione Sepharose 4B.]]> JIN Wei-Lin,LIAO Bai-Song and Jü Gong 1730true Staphylococcus aureus (SA) and cultivated U937 cells by macrophages derived from PBMC was enhanced significantly under lipopolysaccharide(LPS) stimulation. The present experiment showed that the application polyclone antibodies to TLR2 suppressed partly the phagocytosis of macrophages in this model. The LPS-enhanced phagocytosis could be further blocked partly by polyclone antibodies to TRAIL or TNFα. Cells cultivated in a lower serum concentration(1%) was also shown to be a decreased phagocytosis. The results confirmed that TLR2 was a LPS receptor which mediated its signal transduction, while some serum factors participated in the binding of LPS to its membrane receptors. It further suggested that LPS induced effectors such as TRAIL and TNFα might be the key mediators involved in the mechanisms of LPS-enhanced phagocytosis.]]> WANG Liang-Hua,FENG Yu,ZHONG Shan,ZHU Yu-Ping,LOU Yong-Hua and JIAO Bing-Hua 1729true 2-·) produced in PSⅡ particles separated from spinach has been investigated by spin trapping-electron spin resonance (ESR) technique. After bubbling with oxygen and incubating with tetracyanoethylene (TCNE) that acts as an inhibitor of superoxide dismutase (SOD) the experimental evidence of spin adduct of ［DMPO-O2H］· can be obtained by in situ ESR measurements in PSⅡ particles. In contrast, adduct of O2-· and DMPO produced under illumination is obviously decreased when SOD that usually performs as a scavenger of O2-· is present. Furthermore, it is evident to find that the generation of O2-· is positively correlated with the concentration of oxygen. The production of O2-· is also pH dependent, and O2-· concentration reaches maximum when pH is in the range of 6.0～6.5. Either high, or low pH value will lead it to descend steeply.In Tris-HCl washed PSⅡ particles donor-side ET inhibition can causes the production of O2-· decreasing obviously. It can be concluded that the PSⅡ that is active in splitting water and oxygen generation is also a reactive site responsible for the photoinduced O2-· generation in thylakoid of higher plant. In another word, the formation of O2-· is relevantly correlated to the electron transport activity of PSⅡ.]]> LIU Ke,SUN Jian,LIU Yang,ZHANG Qi-Yuan and KUANG Ting-Yun 1728true CUI Yu-Dong,INANAMI O,YAMAMORI T and KUWABARA M 1727true HONG Fa-Shui,WEI Zheng-Gui,ZHAO Gui-Wen,TAO Ye,HU Tian-Dou,LIU Tao and XIE Ya-Ning 1726true 3+ concentration, and used for investigating the transmembrane behaviors of La3+ to PC12 cells. An apparent dissociation constant of La3+-Fura-2 was 3.27×10-11 mol·L-1 in solutions simulating intracellular ionic composition, with pH 7.05. La3+ could not enter PC12 cell under the normal condition, and also could not enter the cell via the calcium channel stimulated by KCl and norpinephrine. However La3+ entered PC12 cell after Na+ within the cell were overloaded using ouabain. Amount of La3+ entering PC12 cell were related to both outer cellular La3+ concentration and intracellular overloaded Na+ concentration. It is suggested that La3+ enter the cell in Na+/La3+ exchanging mechanism.]]> ZHU Wei-Hua,WANG Jin-Xi,LI Hai-Shan and CHEN Yao 1725true HUANG Zhi-Ming,HUANG Kai-Hong,DENG Qing-Li,WANG Wei and SHAO Jing 1724true XIE Jun,HUANG Jing-Fei,SHI Xiu-Fan,SHAO Dan,LIU Shu-Qun,LIANG Chong-Rong and LIU Ci-Quan 1723true MA Zheng-Hai,QIAN Dong,MA Ji,LIN Ren-Yong,WEN Ming,ZHONG Zhe,ZHANG Fu-Chun and ZHANG Qiu-Yun 1722true Petunia by Agrobacterium-mediated transformation. Transformants, especially the ones transformed by αα gene, displayed the obviously higher resistance and accumulation of Pb than control plants. The growth of the roots and shoots of plants transformed by MT and αα genes was unaffected by up to 150 μmol/L Pb and 200 μmol/L Pb, respectively; whereas control seedlings showed severe inhibition of roots’ and shoots’ growth and leaves’ wither under 100 μmol/L Pb. After the transformants were selfed, the seedlings of offspring from MT and αα positive plants were tested for resistance to Pb. It was found that the transformants could tolerate higher concentration of Pb. The detection of accumulation of Pb in the transformed plants by MT and αα genes showed 28% and 35% more than non-transformants, respectively. The results of these experiments indicated that the ornamental plant:Petunia could be used for treatment of the toxic heavy metal Pb pollution in soil.]]> LI Wei,ZHANG Jing,ZHANG Xiao-Yu,SHAN Long and RU Bing-Gen 1721true th chromosome and the predicted exons' sequence of this chromosome is well consistence with the real EST. Thus the RIG1 can be roughly located in 20th chromosome. By use of the exons' sequence predicted from chromosome sequence by GENSCAN, full-length of RIG1 gene has been cloned. Chromosome location of RIG1 gene is further determined by this successful verification of Bioinformatics prediction by experiment. By the same step, genome sequence of RIG1 has been determined. Therefore,by the combined use of Bioinformatics analysis，the full-length cDNA sequence and genome sequence of RIG1 gene are obtained and the predicted protein sequence is determined.]]> LUO Ying,SUI Jian-Li,TIE Yi,ZHOU Ping-Kun and SUN Zhi-Xian 1720true E.coli with the pituitary adenylate cyclase activating polypeptide(PACAP) as an example, the following experiments were carried out. First, on the basis of the reported amino acid sequence of PACAP, DNA sequence of PACAP was deduced and six partially complementary oligonucleotide fragments were designed. The coding region of PACAP was obtained by renaturing the DNA fragments and ligation and identified by DNA sequencing. The coding region of PACAP was cloned into plasmid pGEX-4T-3 and transformed into E.coli BL21(DE3). An expression strain BLPACAP was selected. SDS-PAGE analysis revealed that the GST-PACAP fusion protein was highly expressed and accumulated to about 30% of the total bacterial proteins. By affinity chromatography, up to 90% GST-PACAP was purified by one step from bacterial lysate. The purified protein could promote neurite outgrowth of PC12 cells and the survival of spinal cord neurons.]]> CHEN Zhe-Yu,CHAI Yan-Feng,HE Cheng,LU Chang-Lin and WU Xiang-Fu 1719true TONG Ying-Kai,LIU Hui-Tu and LIU Jun 1718true 118 and Leu119 (called insertion mutant B, InB). The recombinant gene of InB was expressed by Pichia pastoris. The secreted protein was purified by metal chelate affinity and strong cation exchange chromatography. The amidolytic ability of mutant InB is 5 900 IU/mg, the kinetic constants is: Km,plgInB=56.8 μmol·L-1，kcat,plgInB=0.33 s-1. The kinetic constants of plasminogen activation reaction is: Km,plgInB=0.397 μmol·L-1，kcat,plgInB=0.0164 s-1. Fibrin inhibit the catalytiv ability of InB during plasminogen activation, the influence factor is 0.463(means InB remain 46.3% of the catalytic ability when fibrin was involved in the reaction system). The mutant not only has almost the same catalytic ability as wild type scu-PA, but also has strong ability of anti-platelet aggregation(compared with scu-PA), IC50 of InB is 12.7 μmol·L-1.]]> DANG Xin,YANG Jing-Xin,RU Qiang and RU Bing-Gen 1717true Kmfibrin＝4.2 μmol·L-1，greater than the normal Km=0.379 μmol·L-1; kcatfibrin＝0.107 s-1，greater than the normal kcat=0.0165 s-1. The results suggest that existence of fibrin in the reaction system of plasminogen activation depress the affinity between InB and plasminogen, but accelerates the hydrolysis of plasminogen by InB. The count up effect is inhibition.]]> DANG Xin,YANG Jing-Xin,RU Qiang,YUAN Hong-Sheng and RU Bing-Gen 1716true 53 gene (exon7～8) mutatins and p53 proteins and HPV 6，11，16，18-DNA were examined in 49 cervical carcinoma by immunohistochemistry, polymerase chain reaction (PCR) and single strand conformation polymorphism (SSCP) in order to investigate their role and mutual relation and clinical significance in the oncogenesis of cervical carcinoma. The results showed that first, p53 proteins positive rate was 48.98％, and not outstandingly related to the differentiation and the invasive degree of cervical carcinoma(P＞0.05); the defects of P53 gene (exon7～8) were not found but P53 (exon7～8) mutations were detected in 7 of 49(14.29％) cervical carcinoma; then, HPV16-DNA positive rate was much higher than HPV6,11,18-DNA positive rate respectively(P＜0.001),and the different HPV-DNA was simultaneously tested in one cervical carcinoma; last, not all cases of P53 mutations had p53 proteins positive, but the cases of P53 mutations and p53 proteins negative certainly had HPV infections, and HPV positive cases were much more than its negative one in the cases of p53 proteins positive(P＜0.001). These results proved that the oncogenesis of cervical carcinoma is mainly associated with HPV16 infections, and second related to P53 (exon7～8) mutations. p53 proteins positive results from P53 mutations or/and HPV infections in cervical carcinoma.]]> LI Hui-Fang,CHANG Yan-Li,LI Na,YANG Su-Min and JIA Zong-Zhi 1715true I of these components gradually decreased from pH 4.0 according to electrophoresis mobility from higher to lower on PAGE. The molecular weights were in the range of 22～34 ku. 6.5 and 7 were glycoproteins proved by staining with the shiff reagent and thymol/sulfuric acid. The fibrinolytic activity of 7 was highest as determined using chromzym UK and chromzym PL as specific substrates.]]> ZHAO Xiao-Yu and JING Tian-Yu 1714true S.typhi T8-61 LPS or E.coli O111:B4 LPS. After three rounds of biopanning,the clones binding with 2B4 antibody were well enriched with positive rate of 80%. The bindings between 12 of positive phage clones and screening antibody were competitively inhibited by the two kinds of LPS,indicating that the positive clones have similar epitope with LPS. The positive peptide sequences were deduced from the corresponding DNA sequences. There were identical sequences among them. The sequences were GPPQWFFSQPQL （5/12，41.7%），LPQYFWNTATTA （3/12，25%），FPQNHWNVPWAT（2/12，16.6%），HSQSFWNAPLAM and AHPWTHGYFPPL （1/12，8.3%） respectively. The results demonstrate that the peptides screened with 2B4 antibody are mimotopes for LPS conservative epitope.]]> WEN Wei-Yan,HAN Qiang-Tao and FU Ning 1713true o from porcine heart mitochondria. The best results were obtained by the following method: after removing F1-ATPase with NaBr incubation from submitochondrial FoF1-ATPase, Fo was solubilized with CHAPS and purified by sucrose density gradient centrifugation. SDS-PAGE with silver staining showed about 85% purity of the isolated Fo and 9 different subunits including b, OSCP, d, a, e, F6, IF1, A6L and c. The purified Fo was then incorporated into asolectin liposomes, the reconstituted Fo showed higher H+ translocation activity and after Fo was reconstituted with F1-ATPase, the resulted FoF1-ATPase complex exhibited high ATP hydrolysis activity and high sensitivity to oligomycin. The results provide evidence for successful purification, reconstitution of Fo with high H+ translocation activity and its relationship with phospholipids.]]> YANG Hui,ZHANG Ling,ZHANG Xu-Jia and HUANG You-Guo 1712true 3H-TdR incorporation assay. The PKA inhibitor typeⅢ obviously increased the level of 3H-TdR incorporation of S phase in HeLa cells. In contrast with control, the activity of thymidine kinase (TK) in S phase increased, too. It indicated that PKA played an inhibitory role in S phase progression of HeLa cells. With the method of Western blotting, the PKA inhibitor typeⅢ enhanced the level of CyclinA and PCNA, inhibited the expression of p21, which is a negative regulator of cell cycle, but had no effect on the expression of CDK2. The results showed that PKA could negatively regulate the S phase progression by affecting the level of CyclinA, PCNA and influencing the expression of p21 protein. This may be one of the molecular mechanisms which is involved in the negative regulation of S phase progression by PKA in HeLa cells.]]> SUN Xia,LIU Hui-Tu,TONG Ying-Kai and WANG Duan-Shun 1711true cis-trans isomerase (PPIase) activity, similar to the wild type.]]> PEI Wu-Hong,HE Yong-Huai,CHEN Xing,LI Song and SHEN Bei-Fen 1710true WANG Cheng-Xing,LI Xiao-Yan,GU Huan-Hua,DENG Xi-Yun and CAO Ya 1709true Culex pipiens pallens, with a pair of degenerate primers according to the conservative amino acid sequences of CYP4 in insects by RT-PCR. Studies of molecular systematics show that the 24 new genes (alleles) belong to CYP4C,CYP4D,CYP4H and CYP4J subfamilies of the CYP4 family, and they were named by Cytochrome P450 Nomenclature Committee. Among the new genes(alleles),CYP4C23 may be a pseudogene, CYP4H13 has a retained intron 58 nucleotides in length, and CYP4J4V1 has a stop coden(TAG) in frame near the 3′-end.]]> ZHU Chang-Liang,LI Jian-Ming,GAO Xiao-Hong,TIAN Hai-Sheng,LI Xiu-Lan,SHEN Bo and WU Guan-Ling 1708true E.coli. with a level of 30% of the total bacterial proteins. The expressed product was purified by Ni2+-NTA resin, up to 85% purity. The results of activity assays showed that the chimeric protein could significantly promote the survival of spinal cord neurons and had a higher neurotrophic activity than ACTH(4-10) and GDNF respectively.]]> CHEN Zhe-Yu,ZHANG Yong,HE Cheng,LU Chang-Lin and WU Xiang-Fu 1707true E.coli. The Dot blotting and Northern blot showed that this novel gene is highly expressed in the normal adult human brain.]]> DENG Yan-Chun,YAO Li-Bo,LIU Xin-Ping,NIE Xiao-Yan,WANG Ji-Cun,ZHANG Xiao-Guang and SU Cheng-Zhi 1706true －5mol/L in the culture media,the secretion of apolipoprotein AⅠand E increased 36.6% and 49.4%（P＜0.01）respectively, while the secretion of apo AⅡ,B100 and CⅢ decreased 38.9%、31.9% and 29.8%（P＜0.01）respectively.]]> LIU Hao,WU Zhao-Feng and LIU Bing-Wen 1705true ZHANG Chun-Yang,WEI Tao-Tao,MA Hui,DING Yao,CHEN Die-Yan,HOU Jing-Wu,CHEN Chang and XIN Wen-Juan 1704true FAN Yao,WANG Jin,YANG Shan,YANG Xiang,ZHANG Li-Na,HUA Zi-Chun and ZHU De-Xu 1703true 2+-ATPase was purified to nearly homogeneity. The novel feature of this purification is the use of large affinity column and heavy washing to facilitate the purified Ca2+-ATPase with higher activity and protein yield. The specific activity of the purified Ca2+-ATPase was recovered to a maximum of 3.32 μmol·mg－1·min－1 after incubation with asolectin. Silver staining of SDS-PAGE revealed a single protein band around Mr 140 000, showing the purity was over 90%. Different Ca2+ concentrations dramatically affect the specific activity of Ca2+-ATPase.]]> FAN Xiao-Xuan and ZHANG Xu-Jia 1702true in vitro was detected with this method. Using organic solvent (DETC)2-Fe2+-NO complex was extracted from water phase into ester phase, and nitric oxide in sample of large volume can be detected by ESR at room temperature. The extracting ability of several organic solvents such as ethyl acetate, butyl acetate, glycerol triacetate, iso-amy lacetate, and n-butanol, was compared, and it was found that ethyl acetate was a good kind of organic solvent. There was a good linear relationship between the concentration of nitric oxide and ESR intensity within concentration of 20 μmol/L, and the detected limit was improved to below 200 nmol/L; (DETC)2-Fe2+-NO complex is easy to decompose in light but it is very stable in dark at 0～4℃ which, there is only a little change after ten days.]]> ZHANG De-Liang,LI Mei-Fen and ZHAO Bao-Lu 1701true ?-T vector, YA61,one of the overexpressed genes, was analyzed by dot blot and was sequenced then. The sequence gotten was then compared to GenBank data and analyzed by NCBI ORF Finder. Dot blot results showed that the gene YA61 was overexpressed in human gastric carcinoma cell SGC-7901. NCBI's sequence similarity search indicated that the gene YA61 was a new gene sequence. Open reading frame analysis demonstrated that the gene YA61 had one complete open reading frame. In conclusion, the gene YA61 was a new gene sequence that was overexpressed in human gastric carcinoma cell SGC-7901.]]> ZHAO Jin-Rong,YAN Xiao-Jun,HAN Feng-Chan,CUI Da-Xiang,HOU Yu,YAN Quan-Jian and SU Cheng-Zhi 1700true in vitro model was built for researching the effects of strain on vascular smooth muscle cells (VSMCs). The cultured VSMCs was stretched by four-support-bending-beam system, then the project area of cells was measured by computer-image-processing, the adhesion force was measured by micropipette-aspirating system, the α-actin of VSMCs was distinguished by immunocytochemistry and the dynamic of VSMCs was determined by FCM. The results show that: (1) The adhesion force of VSMCs is positively related to time. The adhesion force of unit area is indistinct after stretched for four hour. (2) The amount of α-actin increases with stretching time. (3) The proliferation of VSMCs is a little inhibited by stretched 24 h. These results suggest that the VSMCs in vitro could adjust their behavious to adapt the tension.]]> WANG Hong-Bing,HUANG Qi-Ping,LU Xiao,QIN Jian,WANG Yuan-Liang and CAI Shao-Xi 1699true WANG Cheng,ZHENG Duo,ZHONG Xiang-Yang,XIA Kun,HUANG Liang-Qun,DAI He-Ping,CHEN Yu-Xiang and XIA Jia-Hui 1698true Litchi chinensis sonn. Cv. Heiye) fruit was extracted by phosphate buffer and purified by ammonium sulfate precipitation, ion exchange chromatography using DEAE Sephadex A-50 column, and gel filtration using Sephadex G-100 column. The specific activity of the purified enzyme increased 65.70 fold over the crude extract with 45.66% recovery. The effects of pH and temperature on activity of peroxidase(POD) were assayed. The Km for H2O2 and 4-methylcatechol were determined by Lineweaver-Burk plots. Several compounds including phonelic compounds were used as the substrates of the enzyme for specificity study. The effects of various inhibitors on this peroxidsae were also assayed.]]> GONG Qin-Qin and TIAN Shi-Ping 1697true ZHANG Ai-Qun,CHU Xiang-Yang,LAI Wei-Ling,WU Yan and YEW Tai-Wai 1696true 2+］i (free intracellular Ca2+ concentration) in the target cells. Results showed that full-length HLA-G1 was expressed in the cytoplasm and on the cytomembrane of K562, JAR and CHO cells. HLA-G1 partially inhibited the cytotoxicity of NK92 in a 4h-51Cr-release assay. The ［Ca2+］i in the CHO and GFP-CHO (which expresses the green fluorescence protein) obviously rised after the addition of activated NK92. The expression of HLA-G1 inhibited this kind of rising. These results demonstrated that the rising of ［Ca2+］i in target cells is necessary for the effective cell cytotoxicity. The immune inhibition function of HLA-G1 is possible closely related to the inhibition of ［Ca2+］i in the target cells.]]> TIAN Xing-Hui,ZHOU Jian-Jun,FANG Zhen-Fu,LOU Li-Ming,WANG Yun and FENG Mei-Fu 1695true ZHENG Bin,WEN Jin-Kun,HAN Mei and ZHOU Ai-Ru 1694true YU Cui-Juan,MENG Yan-Ling,GUI Jun-Hao,ZHAO Jing,JIN Ming,WANG Zhi,WANG Cheng-Ji and YANG An-Gang 1693true Helicobacter pylori infection is the major etiological factor of chronic active gastritis and most peptic ulcer disease，and is closely associated with gastric cancers such as adenocarcinoma and MALT lymphoma. Since the Hp adhesin conservative region(AB) is outer membrane protein and porin type component, while these two kinds of protein are the excellent immunogen candidates of vaccination. The gene ab was amplified by PCR and inserted into the prokaryotie expression vector pET-22b（+）and expressed in the BL21（DE3）E.coli strain. DNA sequenced showed one open reading frame of 588 bp,which encoded polypeptides of 195 amino acids. SDS-PAGE and scan analysis show the AB molecular mass is 22.5 ku and recombinant protein amounted to 29% of the total bacterial protein. The AB purity amounted to 96% through affinity chromatography. Western blot analysis of AB confirmed it could be specially recognized by serum from rabbit immunized with AlpA.Acquire of AB established foundation for further studying the molecular adhesin, prevention and cure immunity mechanism of the adhesin.]]> BAI Yang,DAN Han-Lei,WANG Ji-De,ZHANG Zhao-Shan,S.ODENBREIT,ZHOU Dian-Yuan and ZHANG Ya-Li 1692true WU Min-Fan,WU Chun-Fu,ZHANG Hua and TENG Guo-Xi 1691true WU Xing-Zhong and CHEN Ying-Feng 1690true LI Xiao-Qin and LUO Liao-Fu 1689true Autographa california multicapsid nucleopolyhedrovirus) induced Spodoptera litura (Sl-zsu-1) cells apoptosis at 10～12 h post infection, but it could undergo productive replication in Spodoptera frugiperda (Sf-9) cells. The AcMNPV anti-apoptotic gene, p35 gene replication and transcription in different cell lines (Sl-zsu-1 and Sf-9) were investigated by Southern dot blot and RT-PCR respectively. Differences for p35 gene to synthesize and transcribe in these two cell lines suggested that it could not significantly function in the non-permissive cell line due to its untimely and insufficient expression. Inoculation of Sl-zsu-1 cells with wild-type SpltMNPV (Spodoptera litura multicapsid nucleopolyhedrovirus) 12 h prior to AcMNPV infection blocked the apoptosis and rescued the AcMNPV productive replication. It is deduced that P49 protein, a P35 homolog encoded by p49 gene in SpltMNPV genome, might accumulate a certain level at 12 h post infection enough to counteract the caspases effect.]]> ZHANG Ping,YANG Kai,DAI Xiao-Jiang,PANG Yi and SU De-Ming 1688true FAN Heng-Yu,TONG Chao,LI Shi-Wen,CHEN Da-Yuan and SUN Qing-Yuan 1687true 2+-activated potassium (BKCa) channel activity is involved in the pathogenesis of ischemic neuronal injury. Inside-out configuration of patch clamp techniques were used to investigate the temporal changes in BKCa channel activity in CA1 pyramidal neurons acutely dissociated from rat hippocampus at 6 h, 24 h and 48 h following 3 min of brief ischemia. There were no changes in channel unitary conductance and reversal potential after brief ischemia. In contrast, a significant decrease in the channel open probability was observed during the first 24 h following brief ischemia. Kinetic analyses showed that the postischemic suppression of BKCa channel activity was due to a prolongation of the closed time since there was no significant change in open time after brief ischemia. It is suggested that the brief ischemia-induced suppression of BKCa channel activity may be associated with ischemic tolerance.]]> HU Ping,LI Xiao-Ming,LI Jian-Guo,WANG Ying,HUANG Qiao-Bing and GAO Tian-Ming 1686true P＜0.05. The total length of apical dendrite of stress group（155.67 μm±33.32 μm）was significantly shorter than that of control group（195.63 μm±34.61 μm）,P＜0.05. In addition, the ultrastructural changes,including the condensation of the cytoplasm,the reduction of the cell volume,the shrinkage of the nucleus membrane,the degeneration of the mitochodria, and the illegibility of the rough endoplasmic reticulum, were observed in hippocampal CA3 pyramidal neurons in chronic stress rats. It is suggested that the changes of morphology and ultra structure of pyramidal neurons in hippocampal CA3, not in CA1,would underlie the pathophysiology of the stress-related mental disorders.]]> ZHANG Yan-Mei,YANG Quan,XU Chong-Tao,LI Kang-Sheng and LI Wei-Qiu 1685true in vivo by using field potential recording and iontophoresis methods. The field potentials were depressed rigorously by either paired-pulse stimulation or repetitive stimulation at different frequency (10, 20, 50 Hz). Iontophoresis of bicuculline and 2-hydroxy-saclofen decreased the depression degree, while iontophoresis of Ca2+ increased the depression degree. Therefore, the short-term plasticity of synaptic transmission in this pathway is affected by many factors, such as: changing the transmitter release level at pre-synaptic terminal and activity of GABAergic receptors. However, when applying APV, the selective antagonist of NMDA receptors, no any obvious change in short-term synaptic plasticity was obtained. The rigorous short-term plasticity in this visual pathway may play a role in the visual attention function, which is related to the lateral posterior thalamic nucleus.]]> JIA Fan and ZHOU Yi-Feng 1684true 1 hybrid(F1) of HL-type CMS. cDNAs and DNAs of atp6 were obtained from A, B and F1 by PCR and RT-PCR. Then sequences of cDNAs and DNAs are compared: A, B and F1 share the same 15 editing sites found in the transcripts of atp6. The restorer gene in F1 greatly changed the editing frequency of each editing site. So it is suggested that HL-type CMS is associated with RNA editing of atp6.]]> YI Ping,WANG Li,SUN Qing-Ping and ZHU Ying-Guo 1683true in vivo. GFP plasmid was efficiently delivered into muscle by electroporation and robust GFP expression in muscle could be observed more than three weeks. While much less GFP positive cells were observed in tumor and GFP expression could only last 6 days but tumors treated with high voltage/short pulse showed about 2.68 fold more GFP positive cells than tumors treated with low voltage/long pulses. The optimized electroporation parameters was used to mediate therapeutic gene transfer into subcutaneous tumors which derived from T739 mice bladder transitional cell carcinoma cell line (BTT-gfp), human mammary carcinoma cell line (MCF-7) and human hepatoma cell line (SMMC 7721-gfp). Those therapeutic genes included immune reaction regulation factors interleukin12, interleukin2 and GM-CSF or anti-angiogenesis factors such as antisense VEGF121cDNA, soluble form of VEGF receptor (sFlk-1) and Tie2 (ExTek). Inhibition of tumor growth and metastasis were observed in T739 mice carried bladder transitional cell carcinoma and nude mice carried either human breast cancer or liver cancer which were treated with multiple transfers of plasmid encoding interleukine12 mediated by electroporation. MCF-7 and SMMC 7721-gfp derived tumor showed sensitive to single anti-angiogenesis gene therapy, yet definite suppression of growth and metastasis of BTT-gfp tumor was resulted from co-tranfer of sFlk-1 and ExTek gene mediated by electroporation. The results suggest that electroporation is a high efficient, safe and economical method for gene transfer in vivo and electro-gene therapy would be a useful method for solid tumor.]]> WANG Feng,CHEN Xia-Fang,TIAN Yu-Hua,WU Ji-Hong,LI Ling,LI Chuan-Yuan and HUANG Qian 1682true LIAO Wen-Tao,YANG Yang and WU Xiang-Fu 1681true Sulfolobus acidocaldarius was purified to 25 fold by DEAE-Sepharose and Sephacryl S-300 column. The N-terminal amino acids were determined as PGLPKNEHEALEFLKSNNIKWVDLQ, also one consensus sequence of a conserved region TFMPKP(I/L/F)(F/P/Y)(G/R) was found by using alignment of other glutamine synthetase sequences from archaebacteria, a pair of primers were determined according to the above two sequences. The PCR was processed by using S.acidocaldarius genomic DNA as template and a DNA fragment about 780bp PCR product was achieved. After cloning and sequencing of this fragment, the DNA sequence could be translated into a continuous protein sequence which showed high identity to the S.solfataricus GS sequence. After Southern blot of genomic DNA digested by different combination of restriction enzymes using above DNA fragment as probe, a 2.4 kb fragment digested by BamHⅠ/HindⅢ was cloned into pBluescript KS+ plasmid, after colony hybridization, the positive was chosen and a 1.5 kb complete glutamine synthetase gene was sequenced, the gene was then cloned into PET3C plasmide and was induced, the activity of GS was determined. The thermostability of this enzyme showed that it is indeed from thermophilic protein.]]> YIN Zhi-Min,YAN Shu-Zhen,DAI Gu,WU Yi-Fan and ZHANG Shuang-Quan 1680true P＜0.01). A new loading method, step increased stretching was also used. Cells were stretched at 500 με firstly, followed by stretching at 1 000 με for the same hours. Then the strain level was raised to 1 500 με and cells were stretched for the same hours. The step increased stretching experiment showed that osteoblasts absorbed proline to synthesize collagen when received an appropriate signal (500 με). When the strain magnitude was raised, that is to say, the mechanical stimulation was turned into an inhibiting signal, intracellular 3H-proline was released into solution again. These results demonstrate that osteoblasts can adjust themselves to adapt the new mechanical stimulation during the step increased stretching course.]]> TANG Li-Ling,WANG Yuan-Liang,PAN Jun and CAI Shao-Xi 1679true 60Co gamma-ray irradiation mutant Norin 8m. One primer OPG18 produced co-dominant polymorphic markers OPG18/972 and OPG18/943 from Norin 8 and Norin 8m. Based on the clones and sequences of the OPG18/972 and OPG18/943, it shows that a 29 bp DNA fragment in the genome DNA of Norin 8m is deleted. The results directly confirmed the fact that 60Co gamma-ray irradiation can induced a bigger DNA fragment deletion of plant genome DNA.]]> XIANG Tai-He,YANG Jian-Bo,YANG Qian-Jin,ZHU Qi-Sheng,LI Li,NI Da-Hu,WANG Xiu-Feng and HUANG Da-Nian 1678true LIU Rong,WANG Yue-Lan,ZHU Xiao-Peng,LING Lun-Jiang and HAN Ru-Shan 1677true XIONG Wei,ZENG Zhao-Yang,SHEN Shou-Rong,LI Xiao-Ling,LI Wei-Fang,ZHOU Ming,LI Jiang,HE Lin,FENG Guo-Yin and LI Gui-Yuan 1676true E.coli BL21(DE3).An expression strain BLGFRα1 was selected.SDS-PAGE analysis revealed that the rat GFRα1 protein was highly expressed and accumulated up to 21.5% of the total bacterial proteins in the form of inclusion body after the induction. By Ni2+ chelation affinity chromatography, up to 90% GFRα1 protein was purified. Purified and refolded GFRα1 protein could significantly mediate the ability of GDNF to promote the survival and induce the differentiation of PC12 cells.]]> WANG Li-Mei,CHEN Zhe-Yu,ZHU Wei,ZHANG Qing,HUANG Ai-Jun,LU Chang-Lin and HE Cheng 1675true DI)d and the half-time (t0.5) for deformation relaxation of the reticulocytes under the abnormal physiological conditions were measured. (DI)d and t0.5 were respectively put into the reticulocytes membrane shear elastic modulus (E) formula and the membrane viscosity (μm) formula. Through calculation, the value of E and μm in the process of reticulocytes changing into erythrocytes in vivo for 72 h were obtained. It was found that the membrane shear elastic modulus and membrane viscosity decreased in this process of changing. This finding had some basic theoretical and clinical significance.]]> XIE Li-De,LIU Dong-Hai,SUN Da-Gong,YAO Wei-Juan,GU Li,YAN Zong-Yi and WEN Zong-Yao 1674true FENG Yi,LIU Hui-Tu and GAO Ping 1673true GUO Ping,LI Luo-Ye,CAO Yuan-Lin,ZHAO Bao-Lu and LI Zhen-Qi 1672true In vivo, the injection of pNE-mIL12 recombinat plasmid into tumor followed by local X-irradiation one or three times could inhibit the growth of B16 melanoma implanted in C57BL/6J mice and the surviving days of tumor-bearing mice was delayed. It will provide an experimental basis for planning effective clinical gene-radiotherapy of cancer.]]> YANG Ying,LIU Shu-Zheng and FU Shi-Bo 1671true WANG Ji-Shu and HAN Hua 1670true Saccharomyces cerevisiae was treated with nine antifungal agents with known and unknown pharmacological mechanisms. The results indicated that antifungal agents with similar action mode were clustered together. Amphotericin B and nystatin, ketoconazole and clotrimazole have been known to have similar antifungal mechanism respectively. Consistent with their known mechanisms, amphotericin B and nystatin were clustered together; also ketoconazole and clotrimazole were clustered together based on their expression patterns. Solasodine, which was known to inhibit the synthesis of ergosterol, had a close clustering position with ketoconazole and clotrimazole group. Analyzing the relationship among the known and unknown drugs using cluster approach, It can be infered that the pharmacological mechanisms of unknown drugs from their clustering positions relating to the known drugs.]]> ZHANG Liang,ZHANG Yan,ZHOU Yi-Ming,AN Shuang,GUO De-An,ZHOU Yu-Xiang,ZENG Ling-Wen and CHENG Jing 1669true ZHANG De-Li,DING Pei-Guo,LING Lun-Jiang,CHEN Run-Sheng and MA Da-Long 1668true cis-acting element （bases －690 to －850） may be present in mouse PAI-1 promoter. In addition, computer analysis using Match-Search Software found that a new motif of DexRE (dexamethasone response element) 5′ GGTAACCTCTGTTCTCAT 3′ and a putative C/EBPs binding site (cis-motif) exist respectively in the fragment (nucleotides －751 to －770) of, and a sequence (bases －720 to －740) of, mouse PAI-1 promoter,and GMSA and competition assays identified that the trans-acting factors induced by dexamethasone can specifically bind to those cis-motifs. Meanwhile, the site-directed mutagenesis by PCR was performed to detect the influence of mutant DexRE and C/EBPs cis-motif on PAI-1 gene expression. Similarly, the chimeric plasmids containing luciferase as a reporter gene and a fragment of mouse PAI-1 promoter comprising the mutant cis-motifs were constructed, and then transfected into 3T3-L1 preadipocytes. The measurement revealed that the luciferase activities were markedly lowered by mutant DexRE and mutant C/EBPs cis-motif compared with their wild counterparts, implicating that the DexRE and C/EBPs cis-motif identified may control the expression of PAI-1 gene in 3T3-L1 adipocyte. The study is very helpful to elucidate a molecular mechanism through which the dexamethasone may regulate the expression of PAI-1 gene in 3T3-L1 adipocyte.]]> CHEN Ke-Yang,MA Chun-Gu,TANG Qi-Qun and SONG Hou-Yan 1667true YANG Jing,DENG Xi-Yun,TANG Min,WU Shang-Hui,GU Huan-Hua,YI Wei and CAO Ya 1666true 2 cell line (L7) was determined at different time by Western blotting, JNK activation increased with the induction of LMP1 in a time dependant manner was abserved. The expression of phospho-JNK and AP1 activity were analyzed by Western blotting and reporter gene in nasopharyngeal carcinoma cell lines which stably expressing LMP1 and the three kinds of its mutants containing different mutation in carboxyterminal activating region. The result showed that no difference existed between HNE2-LMP1 and HNE2-LMP1ΔCTAR1, but significant difference between HNE2-LMP1ΔCTAR2 and HNE2-LMP1, HNE2-LMP1ΔCTAR1,2. The phospho-JNK expression and AP1 activity of HNE2-LMP1 cell lines were individually transfected by dominant negative TRAF(TRAF-DN) and dominant negative TRADD(TRADD-DN). The results showed that the transfection of TRAF-DN or TRADD-DN made JNK expression and AP1 activity decreased significantly. The results suggested that the functional domain CTAR2 of LMP1 encoded by EB virus can mediate JNK signalling pathway through cooperation with TRAF/TRADD complex.]]> HU Zhi,ZENG Liang,TAO Yong-Guang,TANG Fa-Qing,WANG Hai,LUO Fei-Jun,YI Wei and CAO Ya 1665true LUO Fei-Jun,HU Zhi,ZENG Liang,TANG Fa-Qing,GU Huan-Hua,TANG Min and CAO Ya 1664true in vitro immune library were constructed in parallel conditions, based on the PBLs from healthy and sero-negative blood donors, part of which were in vitro immunized by peptide PreS1 conjugated to BSA. After 3 rounds of panning against PreS1, measurement of antibody-antigen reaction revealed: a scFv specific to PreS1 from the immune library was obtained, which affinity (k=10-7～10-8 M) was higher than that from the naïve one (k=10-6～10-7 M). Sequencing of the two scFv showed they were human antibodies, which may be of interest in therapy of Hepatitis B. This investigation also illustrated that the method of in vitro immunization results in antibody library more satisfied even than the large naïve one.]]> ZHANG Zhi-Chao,HU Xue-Jun,BAO Yong-Ming,YANG Qing,ZHANG Hong-Mei and AN Li-Jia 1663true ZHAO Ping,JIANG Chun-Peng,ZHAO Lan-Juan,WEN Xin-Yu and QI Zhong-Tian 1662true Arabidopsis thaliana, Caenorhabditis elegans and Drosophila melanogaster were studied. It can be concluded that: (1) although the genome sizes and gene numbers are quite different, the compositional complexities of exons are similar, while that of noncoding regions are quite different between eukaryotic genomes. From quantitative perspective, this finding means that the degree of organismal complexity is mainly reflected by noncoding regions, but not by exons; (2) in the same regions of genomic DNAs, composition and compositional complexity are highly similar between chromosomes or contigs within eukaryotic genomes; (3) composition differ remarkably little between introns and intergenic DNAs. This suggests that the effects of transcription, splicing, second structure contribute minimally to the constraints operating on these sequences.]]> LIU Rong,QI Zhen,ZHU Xiao-Peng,LING Lun-Jiang and HAN Ru-Shan 1661true H)-pyridinone (2a) showed the most potent inhibitory activity (IC50=2.52 μmol/L). As removal of substituent at 6- position of pyridinones, the inhibitory abilities of these compounds were almost vanished. The inhibition mechanism of pyridinone derivatives was discussed.]]> XIE Wen-Lin,MA Lin,GUO Gang-Jun,DUAN Zhi-Fang,ZOU Lan,GU Lian-Quan and WANG Jun 1660true Spodoptera litura (SL-1) cell transfected with DNA of Spodoptera litura nucleopolyhedrovirus revealed progressive cell blebbing starting at 6 h postinfection and culminating in total cell destruction at 18 h postinfection. The fragmentation of the infected cell nuclei and apoptotic body were observed by stained with the specific fluorescent dye DAPI. Agarose gel electrophoresis analysis of the DNA extracted from infected cells showed typical DNA ladder. All these supported that the transfected SL-1 cells undergo apoptosis. Neither apoptosis nor polyhedra was observed in the SL-1 infected with the virions of SpltNPV.]]> DU Chang-Sheng,PENG Jian-Xin and HONG Hua-Zhu 1659true 13 particles/ml and 1.08×1013 particles/ml respectively. The particles of single gene rAAV /ATR or rAAV /p21 were 6.54×1012 particles/ml or 1.06×1013 particles/ml.]]> MENG Xian-Min,WU Xiao-Bing,MI Li-Guo,WU Zhi-Jian,ZHAO Xiu-Wen,LIU Dong-Qing and DING Jin-Feng 1658true LI Hong,WANG Meng-Wei,WANG Gang-Shi,CHEN Run-Sheng,LING Lun-Jiang and WANG Jin-Hua 1657true Lumbricus bimastus）with the molecular mass about 30 ku was extracted by SDS-PAGE. Then the N-terminal of the protein was sequenced. The PCR primer was designed according to the N-terminal sequence, and its cDNA fragment was obtained by RT-PCR. The cDNA was cloned into pGEMT-vector and sequenced. The sequence showed that the fragment was 888 bp,with the ORF of 726 bp and the 3′ untranslation terminal area. The ORF encoded a protein of 242 amino acids, so the protein was named as PV242.The prediction of protein structure shows that the PV242 has two domains，between which is the active sites-His44 and Ser191.At the same time, the pI 4.33 of PV242 was detected, and it is a protease in trypsin family of serine protease superfamily. PV242 was expressed in pTrxFUS expression system as the fusion protein TrxA-PV242 in soluble form, the TrxA was a molecular chaperon which made the expressed protein soluble. The fusion protein was purified by ion exchange chromatography. The purified fusion protein TrxA-PV242 has fibrinolytic activity.]]> XU Yi-Hui,LIANG Guo-Dong,SUN Zhao-Jun,CHEN Fei,FU Shi-Hong,CHAI Yu-Bo and HOU Yun-De 1656true bis(7)-tacrine can prevent cell apoptosis induced by staurosporine in NG108-15 and HeLa cell lines. Phase-contrast and fluorescence microscopes were used to examine cell morphological changes. MTT assay was used to examine if staurosporine impairs cell metabolism. DNA was isolated and electrophoretically separated on 1% agarose gel to observe if there were DNA fragments. Western blot was made to analyse protein levels of anti-apoptotic Bcl-2 and proapoptotic Bax. The results showed that NG108-15 cells treated with 0.1 mol/L staurosporine for 12～24 h exhibited marked cell death and DNA fragmentation. Pre-treatment with 0.1 μmmol/L tacrine provided approximately 40% protective effect and resulted in obvious inhibition or delay DNA fragmentation. Moreover, NG108-15 cells treated with tacrine became elongated and polarized, and showed longer processes than control cells. Pretreatment with 0.1 mmol/L tacrine significantly increased the expression of Bcl-2 protein level and delayed the staurosporine-induced increase of Bax protein expression. However, bis(7)-tacrine did not show any protective effect on the cell impairment induced by staurosporine in NG108-15 cells. In HeLa cells 0.1 μmol/L staurosporine also induced significant cell injury, but pretreatment with tacrine and bis(7)-tacrine did not provide any obvious protective effect against this cell damage. These data suggest that tacrine markedly protect against apoptosis of NG108-15 cells but bis(7)-tacrine did not. Both tacrine and bis(7)-tacrine were not show to inhibit apoptosis of HeLa cells. It appears that the protective effects of tacrine against apoptosis not be mediated through AChE inhibition and has cell selective specifity.]]> PENG Fang-Fang,ZHANG Bai-Fang,ZHANG Jiang-Zhou and WU Dong-Cheng 1655true 20 were detected with Western blotting assay. The results showed that during the process of DA-induced apoptosis, the cleaved caspase-3 P20 and the release of cytochrome c into cytosol were elevated markedly compared with the controls. The protein levels of Bcl-2 were decreased, however, Bax protein was increased significantly. After pretreatment of PC12 cells with cyclosporine A at 1 μmol/L for 24 h, the release of cytochrome c and caspase-3 activation were blocked almost. But it didn't show effect on protein levels of Bcl-2 and Bax. The results suggested that Bcl-2 family and cytochrome c and caspase-3 might be involved in the progression to apoptosis induced by DA, furthermore, the release of cytochrome c from mitochondria might be an essential event.]]> ZHU Yuan-Gui,CHEN Xiao-Chun,CHEN Li-Min,FANG Fang,ZHOU Yi-Can and ZHAO Chao-Hui 1654true in vivo antiangiogenesis experiments.]]> ZHANG Hua and SHOU Cheng-Chao 1653true E.coli, The coding region with SalⅠ and NotⅠ restriction sites of BRD7 was obtained from pGEM-T Easy/BRD7 plasmid by PCR. PCR product and plasmid PGEX-4T-2 were digested by corresponding restrict endonucleases respectively. The fragments were ligated by T4 DNA ligase to gain recombinant expression vector. Endonuclease digesting and DNA sequencing confirmed that the coding region of BRD7 gene was correctly inserted into the vector. The recombinant plasmid PGEX-4T-2/BRD7 was transferred into competent Jm105 strain. The GST/BRD7 fusion protein was expressed in the bacteria under induction of IPTG. After induction, a new protein band of 90 ku appeared on SDS-PAGE. The result was confirmed by Western blot. The recombinant protein of 90 ku amounted to 28.48% of the total bacterial protein after inducing with IPTG for 4 h at 37℃. It existed not only in supernatant but also in precipitation of broken bacteria. The successes in construction of expression vector of BRD7 and expression of BRD7 in E.coli make it possible to study further on its biological function and antibody preparation.]]> NIE Xin-Min,ZHANG Bi-Cheng,XIANG Juan-Juan,ZHU Shi-Guo,ZHOU Ming,DONG Li,YU Ying,LI Xiao-Ling and LI Gui-Yuan 1652true ND-4L, ND-4, COX-2, Lys-tRNA, ATP-8 and ATP-6 were identified from 69 SSH positive clones. The results indicated that NO can distinctly induce overexpression of mitochondria DNA encoded genes in the esophageal carcinoma cells. In addition, it was discovered that there were single nucleotide substitution in three sites of the fragment of ND-4L and ND-4 genes (10 736～11 449, 10 872 T→C, 11 001 A→G, 11 346 A→G), and one single nucleotide deletion (8 380，A) which will lead to occur the frame-shift mutation in the peptide in the fragment of COX-2/Lys-tRNA /ATP-8/ATP-6 genes (8 011～8 589). The analysis of amino acid sequences showed that an incorrect structural ATP-8 subunit existed possibly in the EC109 cell induced by NO. These results provided a new clue for further exploring the mechanism of NO effecting on carcinoma cells.]]> LI En-Min,XU Li-Yan,Yang-Fan,YUAN Lan and CHEN Yue 1651true 1 protein expression in multistage tissue of pathological changes during the nasopharyngeal carcinogenesis and nasopharyngeal carcinoma(NPC), the simple hyperplasia/ metaplasia, atypical hyperplasia/metaplasia in the nasopharyngeal paracarcinoma mucous epithelium and NPC were studied for Cyclin D1 expression with immunohistochemical streptavidin-peroxidase (SP) method by tissue array techenique. The positive rates were 30.0%（3/10）,90.0%（18/20）and 62.9%（39/62）respectively,among which hightening “instantaneously” in the atypical hyperplasia/metaplasia appeared. It is shown high Cyclin D1 expression may be an early incident in the course of nasopharyngeal carcinogenesis, and that atypical hyperplasia/metaplasia probably be an important “toll-gate”.]]> SONG Xin,ZHAO Xiao-Rong,WANG Yi,ZHOU Jian-Hua,GUAN Xin-Yuan and CAO Ya 1650true Gloydius shedaoensis) venom gland. The 711 bp nucleotide sequence, which encodes the mature gloshedobin, was cloned into expression vector pPIC 9K and transferred into yeast Pichia pastoris, strain GS115. Transfermants with phenotype His+Mut+ were selected to study their expression. Recombinant protein was conveniently separated and purified from the supernatant by two chromatographic steps: ion exchange chromatography on Q-Sepharose FF and affinity chromatography on Benzamidine-Sepharose 4BCL. Like intact gloshedobin, the recombinant enzyme exhibited strong esterase activity using tripeptide p-nitroanilide derivatives as substrate, but hydrolyzed N-p-tosyl-L-arginine methyl ester (TAME) very weakly. The recombinant protein displayed extreme instability at 37℃ in neutral buffer but higher stability at 0℃, and also, pH is not a key factor to affect its stability while the optimal pH for its enzymatic activity is pH 8.0.]]> YANG Qing,HU Xue-Jun,XU Xiao-Ming,GAO Xiao-Rong,AN Li-Jia,SU Zhi-Guo and J.C.JANSON 1649true QIAN Jun-Min,LI Xu-Xiang and SUO Ai-Li 1648true 2+ and SeO32- against the changes were studied. The results showed that these changes can be partly antagonized by 1 mg/L Na2SeO3 and completely antagonized by 5 mmol/L Mg2+.]]> ZHANG Xu,LI Xiao-Bing,LI Sheng-Guang,JIANG Pei-Dong and LIN Zhi-Huan 1647true TM Mouse cDNA array 1.2 in Babl/c mice lung tissues during endotoxic shock. Results showed that the expression of 128 genes was up-regulated and 3 genes down-regulated at 2 hours after exposure to endotoxin; the expression of 51 genes was up-regulated and 21 genes down-regulated at 20 hours after exposure to endotoxin. The changes in gene expression were further confirmed by RT-PCR. The significance of above changes in gene expression was analyzed. The results would be helpful to elucidating the molecular mechanism of endotoxic shock.]]> YUAN Kai-Yu,ZHAO Zhen-Yu,LIU Ying,ZOU Jiang,LIU Mei-Dong,CHEN Guang-Wen,YOU Jia-Lu and XIAO Xian-Zhong 1646true YANG Qing-Wu,ZHU Pei-Fang,WANG Zheng-Guo and JIANG Jian-Xin 1645true SHI Xiu-Fan,HUANG Jing-Fei,LIU Shu-Qun and LIU Ci-Quan 1644true 5 PFU per mice. A good foundation has been laid down for developing PRV genetically-engineered subunit vaccine.]]> CHEN Xin-Hua,YANG Lin,LONG Qi-Xin,WANG Xun-Zhang,CHEN Qu-Hou,HONG Wen-Zhou and CHEN Huan-Chun 1643true E.coliBL21(DE3) harboring plasmid PET22b-mETIa were induced by IPTG and then recombinant ETIa were highly expressed.Expressed rETIa were above 40% of total bacterial protein.After primary purification through breaking E.coli, dissolving inclusion bodies, refolding,and further purification by two-step chromatographies, rETIa of electrophoretic purtity has been obtained.Inhibitory activity of rETIa against tPA deletion variant (NTA) has been detected and inhibitory constant (Ki) was 8.72×10－8mol/L.So affinity chromatography column of rETIa-Sepharose 4B was prepared for purification of NTA.After only one-step purification with this column from refolded NTA,13.2-fold puritied NTA with the specific activity of (565.7±71.3) U/μg and above 90% of purity,have been obtained with the recovery rate of 96.2%.]]> WANG Xin,QU Xian-Ming and YANG Sheng-Li 1642true Inimicus japonicus was constructed. The cDNA was cloned into eukaryotical expression plasmid pcDNA3.0. SMARTTM protocol was used for cDNA library construction and bioinformatics analysis was carried out. 94 novel EST clones were obtained from 150 sequences in the library, of which there were 35 full-length clones, including cytolysin genes, short neurotoxin gene, C-type lectin gene, macrophage migration inhibitory factor gene and so on. Most of those genes were reported for the first time in Inimicus japonicus. Further studies on those genes and large scales of sequencing in the library are going on, which may be helpful to make clear the components of Inimicus japonicus venom and understand the function of those proteins at molecular level.]]> JIANG Xiao-Yu,TU Hong-Bin,CHEN Hui-Ping,WEI Jian-Wen,YANG Wen-Li,WU Wen-Yan and XU An-Long 1641true XU Dong-Gang,MA Bai-Kun,LI Li,ZOU Min-Ji,PENG Shan-Yun and WANG Jia-Xi 1640true Osrab5B，designated for rab5B gene from rice, encodes a putative Rab protein. In order to investigate the physiological and biochemical functions of this protein OsRab5B, the complete coding sequence (CDS) of Osrab5B was amplified and inserted into a prokaryotic GluGST expression vector, pGEX-4T1, thus constructing the prokaryotic expression vector. The in-frame recombinant plasmid was confirmed by sequencing and was named pG-5BE. Then the vector pG-5BE was transformed into prokaryotic cell E.coli strain BL21 (DE3). The GST fusion protein expressed by this positive strain was purified with GSTrapTM column. The result from GTP-binding assay proved that the fusion protein has GTP-binding ability.]]> LIN Hui-Xian,LIU Xiao-Bin,LIANG Cheng-Ye and LIU Liang-Shi 1639true TM Rat cDNA Expression Array. The difference of gene expression profiles was analyzed using image analysis instrument. And 15 differential expression genes were discovered each in hippocampi and cerebral cortex. In the hippocampi, 12 genes were high expression in the P77PMC rats but low expression in the normal Wistar rats, 3 genes were high expression in the Wistar rats but low expression in the P77PMC rats.In the cerebarl cortex,13 genes were high expression in the P77PMC rats but low expression in the normal Wistar rats, 2 genes were high expression in the Wistar rats but low expression in the P77PMC rats. Thus, there are several differential expression genes between P77PMC rat and Wistar rat. All these differentially expressed genes may play important roles in the pathogenesis of epilepsy.]]> WU Zhi-Guo,XIAO Bo,YANG Xiao-Su,LI Guo-Liang and XIE Guang-Jie 1638true 121 cDNA was amplified by RT-PCR, and was inserted into the Pichia pastoris expression vector pPIC9K to form the expression plasmid p9KVEGF121. This recombinant plasmid was transformed into GS115. Transformants were screened by G418-YPD plates and were induced by methanol. The expression product of r-hVEGF121 amounted to 900 mg/L by a 5-liter fermentor, over 70% of the total secreted protein. The purified r-hVEGF121 can stimulate the proliferation of bovine capillary endothelial cells and shows high vascular permeability.]]> WANG Yue-Xiang,GUAN Xiao-Qun,YANG Jian,MO Wei and SONG Hou-Yan 1637true HAO Mei-Rong,YANG Ming and BU Xian-He 1636true YUAN Zhong-Hua,YANG Yong-Zong and YANG Xiao-Yi 1635true LI Jian,LIU Xin-Ping,LIN Shu-Xin,DENG Yan-Chun,MENG Qing-Jun,ZHANG Wen-Hong,LI Shu-Jun,NIE Xiao-Yan and YAO Li-Bo 1634true FU Yan,DENG Wei-Guo and YANG Xiao-Li 1633true WANG Jie-Ru,QIAN Jun,DONG Li,LI Xiao-Ling,TAN Chen,LI Jiang,ZHANG Bi-Cheng,ZHOU Jie and LI Gui-Yuan 1632true 7 and baculovirus transfer vector pFASTBac. Glycophorin A and band3 C-terminus were co-transformed into the yeast strain AH109 and the interaction was identified by nutritional selection and β-glycotosidase activity detection. The GPA expression product in Sf9 cells was analyzed on Western blot with anti-GPA and anti-human band3 antibodies, and the result showed that they have immunologic cross-link reaction. All the results above confirmed the interaction between band3 and glycophorin A.]]> DU Hong-Qing,FU Guo-Hui,JIANG Xiao-Shu,LONG Xiao-Yu,ZHANG Zhen-Yu,YANG Bao-Feng and KONG Xian-Gang 1631true in vitro and in vivo were analyzed. This sequence showed strong anticancer activity in vitro and in vivo and had dose -dependent relation. The sequence had no obvious toxicity on tumor -burdended nude either . IGF1R could be used as an appropriate target for tumor therapay.]]> LIN Li,WANG Sheng-Qi,GUAN Wei,YANG Bing-Hu and HU Xiao-Dian 1630true 2+］i) rise and the inhibiting effect of selenium on this process were determined by using Fura-2/AM fluorescence measurement method in a human umbilical vein endothelial cell line: ECV-304. Experimental results indicated that there was a fast rise of ［Ca2+］i by the treatment of S-nitrosoglutathione(GSNO), a donor of NO. The rise of ［Ca2+］i was inhibited by Hb, a scavenger of NO. These results suggested that this rise of ［Ca2+］i was induced by NO. There was no effect on ［Ca2+］i response to NO by removing calcium ion from bath, or by adding the non-selective calcium channel antagonist, CdCl2（1 mmol/L）to the bath. When the cells were pretreated by sodium selenite(1 μmol/L),the rise of ［Ca2+］i induced by NO was obviously inhibited. This result suggested that selenium can inhibit calcium store release.]]> DENG Ying,HUANG Kai-Xun and XU Hui-Bi 1629true ZHOU Yi-Feng,YANG Yu-Peng,HU Bing and SHOU Tian-De 1628true in vivo and in vitro were measured. The coincidence of the results in vivo and in vitro demonstrated the correctness of the method. As expected, the photobleaching rate increased near-linearly with the excitation power for one-photon excitation. However, the two-photon photobleaching rate increased with high-order power (≥3.5) of excitation power, indicating the presence of high-order photon interaction in two-photon excitation microscopy. The same results are obtained by photobleaching experiments of the green fluorescence protein. As a consequence, the use of multi-photon excitation microscopy in the study may be limited by increased photobleaching.]]> CHEN Tong-Sheng,ZENG Shao-Qun,LUO Qing-Ming,ZHANG Zhi-Hong and Zhou Wei 1627true ZHANG Jing and SHI Xiu-Fan 1626true ZUO Lian-Fu,LIN Pei-Zhong,QI Feng-Ying,ZHANG Lin-Xi,GUO Jian-Wen and LIU Jiang-Hui 1625true KONG Ying,GE Chang-Hui,YAN Qiu and ZHU Zheng-Mei 1624true 2O2 treatment in an ATP-containing buffer at 37℃.It was observed under fluorescence microscope that FITC-labeled MTS-αB-C had gone into neonatal rat cardiomyocytes by MTS mediation after the cells were incubated with the MTS-αB-C for 8 hours.]]> JIANG Lei,LIU Shuang,YUAN Can,XIAO Wei-Min,WANG Kang-Kai,YOU Jia-Lu and XIAO Xian-Zhong 1623true ZHANG Zhi-Hong,ZENG Shao-Qun,LUO Qing-Ming,LIU Ya-Feng,ZHOU Wei and CHEN Tong-Sheng 1622true Colletotrichum gloeosporioides was modeled. The predicted structure is similar to those of its homologous MT proteins in metal-Cys combination, and reasonable in its structure energy as well as in its fold motif.]]> HE Hong-Zhen,Lü Tun,ZHAO Nan-Ming,ZHANG Ri-Qing and LIU Jin-Yuan 1621true Saccharomyces cerevisiae cells in water/organic solvent biphase system was studied. The effects of shake speed, the hydrophobicity of organic solvent, volume ratio of water phase to organic phase, pH of aqueous phase and reaction temperature on the initial reaction rate, maximum yield and enantiomeric purity of the product were systematically explored. All the factors mentioned above have important effects on the reaction. n-Hexane is found to be the best organic solvent for the reaction. The optimum shake speed, volume ratio of water phase to organic phase, pH and reaction temperature are 150 r/min, 1/2, 8 and 25～30℃ respectively for the reaction, under which the maximum yield and enantiomeric purity of the product are as high as 96.8% and 95.7%(ee) respectively.]]> LOU Wen-Yong,ZONG Min-Hua,FAN Xiao-Dan,LU Jie-Quan and DU Wei 1620true XIANG Juan-Juan,YU Ying,WANG Jie-Ru,ZHU Shi-Guo,ZHANG Bi-Cheng,LI Zhong-Hua and LI Gui-Yuan 1619true Burset/Guigò test data set.]]> YANG Wen-Qiang,QIAN Min-Ping and HUANG Da-Wei 1618true ZHANG Hai-Qian,S.HIROAKI,GU Ning,S.HIROSHI and S.MASAHIKO 1617true LIU Zhe,CHEN Yun,YANG Ying and PENG Jing-Pian 1616true +8+ DP thymocytes lessen but CD4-8+ thymocytes increase. And as the avidity of DP thymocytes with sites of TSCs increases, more DP thymocytes differentiate into SP thymocytes. How the thymic stromal cells mediate the differentiation from DP thymocytes to SP thymocytes is described.]]> ZHANG Li-Mei and DU Chan-Ying 1615true 3+，Cd2+ and Pb2+ could influence the activity of amylase from porcine pancreas and competitively replace Ca2+ from amylase. The results showed that the activity of amylase was enhanced under the treatment of Ce3+，Cd2+ and Pb2+ respectively at low concerntration, but inhibited at the high concerntration. Both replacement and inhibition showed that the action was in the order: Pb2+＞Cd2+＞Ce3+.]]> HONG Fa-Shui,WANG Xue-Feng,SHEN Song-Dong,SU Guo-Xing and PAN Xin-Fa 1614true + channels play an important role in coupling membrane excitability with intracellular metabolic stress. To characterize such KATP channels from rat brain, the inside-out mode of patch-clamp technique was applied to freshly dissociated hippocampal CA1 pyramidal neurons of adult rat. One type of K+ permeable channel was recorded only at the presence of Ca2+ in the internal solution and it could be inhibited by application of 1～3 mmol/L ATP and 1 mmol/L tolbutimade, a KATP channel blocker. Both of channel open probability and the ATP induced-inhibition displayed a voltage-dependent fashion. When both sides of the excised membrane were in symmetrical 140 mmol/L K+, the I-V relation was linear with a conductance of 204 pS and reversal potential was 3.57 mV. Unlike the previously reported “classical” KATP channel, this large-conductance KATP channel (L-KATP) was regulated by membrane potential, intracellular Ca2+ and ATP, indicating a new subtype of KATP channel presents in hippocampus neurons. These results demonstrate that at least two distinct KATP channels exist in rat hippocampal neurons and suggest that metabolic state may be continuously sensed in neurons via different KATP channels with resulting alterations in neuronal membrane excitability.]]> ZHOU Ying-Jie,TONG Zhen-Qing and GAO Tian-Ming 1613true WANG Wei,WU Jun-Feng,ZHANG Zong-Yu and TONG Tan-Jun 1612true Mycobacterium tuberculosis was shown to be a trimer-of-trimers. Hsp16.3 proteins were denatured under three kinds of strong denaturing conditions by heat treatment (at 100℃, 15 min) or chemical reagents (12 mol/L urea or 8 mol/L guaridine, 4 hours) and then were renatured by cooling or dialysis. The secondary, tertiary and quaternary structures of Hsp16.3 were investigated by using far- and near- UV circular dichroism as well as pore-gradient polyacrylamide gel electrophoresis, respectively. The data clearly showed that the renatured Hsp16.3 proteins almost completely regained its native conformation, thus suggesting the strong ability of Hsp16.3 to refold and reassembly to its native conformation.]]> MAO Qi-Long,FENG Xiu-Guang and CHANG Zeng-Yi 1611true Zea mays L.) leaves produced significant changes under illumination. With the raising of light intensity, zeaxanthin(Z) content evidently rose, violaxanthin(V) content significantly decreased, antheraxanthin(A) content significantly increased in low and middle level lights, and then decreased slightly in strong lights. The xanthophyll cycle components pool size V+A+Z only enlarged a little. Under the same conditions，the non-radiative energy dissipation of leaves apparently enhanced，which was shown that non-photochemical quenching (NPQ) rapidly rose，while Fv/Fm apparently reduced. It was analysed that (Z+0.5A)/(V+A+Z) had exhibited apparently positive linear correlation with NPQ，and showed a negative correlation with Fv/Fm. Compared to (Z+0.5A)/(V+A+Z), (V+0.5A)/(V+A+Z) had showed contrary results. Based on the results, it was deduced that Z epoxidation and V deepoxidation had been significantly related to non-radiative energy dissipation and PSⅡ light energy conversion in detached maize leaves.]]> DONG Gao-Feng,YANG Cheng-Wei,XU Zhi-Fang and CHEN Yi-Zhu 1610true ZHANG Fang-Lin,LI Guo,YE Chuan-Zhong,DING Wei,LIU You-Ping,XIE Chao,ZHANG Di,SUN Wei-Hua and LUO Min 1609true TAO Ye and ZHANG Jing-Pu 1608true Escherichia coli. The purity examined by SDS-PAGE showed as a single band. Its molecular mass measured by mass spectrum and the amino acid sequence of N terminal were in consistence with CspC, one of the cold shock proteins, in E.coli. Subsequently, the content of its secondary structure was estimated from the circular dichroism spectra, and moreover, its stability in high temperature and the conformational change after binding with single strand DNA were monitored with a CD spectrophometer.]]> LIU Wei,LI Yiao,XIANG Ye and WANG Da-Cheng 1607true CHEN Shan,LIU Dong-Bo,XIA Hong-Mei,HE Meng-Yuan,HAO Shui,ZHUANG Yu-Gang,AN Yu-Xian and DONG Li-Song 1606true cis-acting elements were scanned within 11.5 kb of the 3′juxtaposed region of the Yunnanese (Aγδβ)0-thalassemia deletion using the luciferase report gene system. A 1.7 kb fragment immediately downstream of the 3′ breakpoint of the deletion was found to increase expression of the luciferase gene driven by Gγ-globin gene promoter by 3.8 to 4.0 fold in K562 cells and MELGM979 cells and 1.5 fold in HeLa cells. A 1.4 kb fragment that is located 10 kb downstream of the 3′ breakpoint was found to increase the luciferase expression by 2.4 to 2.9 fold in K562 cells and MELGM979 cells but no enhancement in HeLa cells. The results suggested that the two fragments contain enhancer-like elements and they function in a certain erythroid-specific manner. Furthermore, a 430 bp region that contains several putative motifs for known transacting factors binding within the 1.7 kb fragment was showed to include the most of the enhancer activity of the fragment. These results provided experimental proof for the hypothesis that the importation of enhancer-like sequences into the vicinity of Gγ globin gene may be responsible for the reactivation of the Gγ-globin gene in the Yunnanese (Aγδβ)0-thalassemia mutant.]]> HUANG Xiao-Dong and ZHANG Jun-Wu 1605true E.coli and to explore whether it has the biological effect on the inhibition of angiogenesis, a 630 bp of MMP-2 C-terminal coding fragment (PEX) was cloned into prokaryotic expression vector pCal-n from primary cultured day 10 chicken embryo fibroblasts using RT-PCR. The relatively pure inclusion bodies of CBP/PEX fusion protein were denatured and renatured by guandine·HCl method from PEX/pCal-n plasmid transformed log phase E.coli BL21 (Des) pLys induced with IPTG. Growth curve of human umbilical vein endothelial cells (HUVEC) and chick embryo chorioallantoic membrane (CAM) assay were performed to examine the effects on the inhibition of angiogenesis. The prokaryotic expressed CBP/PEX fusion protein has the biological activity and can inhibit the growth of HUVEC in vitro and the angiogenesis on chick CAM in vivo. These results suggest that the expressed PEX is to be a novel potential therapeutic inhibitor of diseases associated with angiogenesis.]]> LI Jin-Ping,ZHANG Guang-Mou,KE Yang,LIN Ben-Yao,ZHAO Wei,HU Ying,NING Tao,LIN Zhong-Xiang and ZHANG Zhi-Qian 1604true LI Xiao-Qin and LUO Liao-Fu 1603true P＜0.01) when E10 embryos were rotated for 4 h, and KCl- induced release rate and release content from brain cells of E10 embryos rotated for 24 h were higher than those from controls. However, there was no significant difference in the total intracellular glutamate concentration and the KCl- induced release rate, release content from brain cells of E13 embryos between rotated and their control groups. These results suggest that the releasing behaviors of neuro-transmitters from brain cells can be affected by microgravity and the influence is related to the age of the embryos. The dynamic process of glutamate release from brain cells induced by a singlee electric pulse shows that the release is related with a rapid increase of the intracellular ［Ca2+］i following the pulse.]]> WANG Yan,SUN Tong and ZHANG Jin-Zhu 1602true DENG Qing-Li,HUANG Kai-Hong,SHAO Jing,HUANG Zhi-Ming and WANG Wei 1601true Lactuca sativa L.) chloroplasts. The results showed that the production of superoxide anion increased photochemical quenching(qP) and non-photochemical quenching(qN) evidently. As superoxide dismutase (SOD) was inhibited by sodium diethyldithiocarbamate (DDC) in chloroplasts, qP decreased and qN increased in the quenching of chlorophyll fluorescence induced by X+XO, while the increasing extent of qP was not great and qN rose insignificantly in the quenching of chlorophyll fluorescence induced by MV. When iodoacetamide (JAA) inhibited carbon assimilation, qP decreased and qN increased. Uncoupler NH4Cl promoted the permeability of across thylakoid membrane proton, which made qP rising and qN decreasing. Under uncoupling conditions, MV raised qP and qN insignificantly. It was analyzed that the producing and scavenging of superoxide anion in time was important to maintain photosynthetic electron transport and to improve transmembrane ΔpH. It contributed to the conversion and dissipation of photon energy absorbed in chloroplasts, and reduced the harm of photoinhibition to a certain extent led by excess photon energy.]]> XU Zhi-Fang,LUO Guang-Hua,KE De-Sen,CHEN Jin-Ting,CHEN Yi-Zhu and WANG Ai-Guo 1600true WANG Ji-Shu,YANG Xi,LI Rong,ZHOU Peng,ZHANG Miao-Li and HAN Hua 1599true cpcβ gene which encodes the β-subunit of phycocyanin and the 1.4 kb fragment of glnA gene encoding glutamine synthetase were obtained by polymerase chain reaction (PCR) from genomic DNA of marine cyanobacterium Synechococcus sp. PCC 7002. Then, integrative expression vector pKGC-MT, which contained promoter Pcpcβ, mMT-Ⅰ gene and integrative platform glnA, was constructed and introduced into Synechococcus sp. PCC 7002 via natural transformation. Selected by ampicillin, the stable transgenic cyanobacterium was obtained. PCR analysis indicated the integration of mMT-Ⅰ gene in genomic DNA of Synechoccus sp. PCC 7002 and Western blotting demonstrated the expression of mMT-Ⅰ in the cyanobacterium. According to the result of ELISA, the amount of the expressed mMT-Ⅰ was about 800 μg/g fresh cells.]]> ZHOU Jie,LUO Na,NING Ye,SHI Ding-Ji,YU Mei-Min and RU Bing-Gen 1598true in vitro.]]> HAN Ya-Ling,KANG Jian,ZHANG Jian and LI Shao-Hua 1597true n=10). The channel open probability was voltage dependent (Vm=－60 mV，Po=0.44±0.12；Vm=+60 mV，Po=0.86±0.06, n=10). The reversal potential of the channel was (－4.17±1.84) mV in symmetrical Cl－ concentration (150 mmol/L). When partially substituting Na-Gluconate for NaCl, the reversal potential shifted to (－34.23±4.86) mV (［Cl－］i/［Cl－］o=(30 mmol/L)/(150 mmol/L)). This shift indicates a high Cl－ selectivity of the channel. Chloride channel blockers DIDS and SITS showed a reversible inhibiting effect on the channel activity. These results provide a report of an outwardly rectifying chloride channel (ORCC) in hippocampal CA1 pyramidal neurons of adult rat.]]> LI Jian-Guo,LI Xiao-Ming,HU Ping,WANG Ying,LI Xiao-Wen,QIAO Jian-Tian and GAO Tian-Ming 1596true ZHANG Shao-Wu,PAN Quan,CHEN Run-Sheng and ZHANG Hong-Cai 1595true 2 for 15 min at 37℃. The findings were that Ca2+ treatment at concentration 1～5 mmol/L led to significant proteolysis of tau protein and this degradation was blocked by calpain inhibitor, calpeptin. In addition, when the extracts containing 1 mmol/L CaCl2 were treated with μ-calpain inhibitor (0.05 μmol/L of calpastatin) or m-calpain inhibitor (100 μmol/L calpain inhibitor Ⅳ) or both, the Ca2+-induced degradation of tau protein was decreased to 8.6%，92.5% and 97.8%, respectively. These data suggest that both μ-calpain and m-calpain in brain cortex extracts are activated by Ca2+ and both of them degrade tau protein, although, m-calpain plays a more important role in proteolysis of tau.]]> FANG Zheng-Yu,LIU Shi-Jie,WANG Xiao-Chuan,LIU Rong,WANG Qun,CHEN Zheng-Yue and WANG Jian-Zhi 1594true SUN Xiao-Yang,TAN Yin-Fei,TANG Shuang and WANG Yan-Ling 1593true CHEN Xiang-Gui,LI Yong,ZHANG De-Li,CHENG Jun,ZHU Wen-Li and DAO Jing-Jing 1592true HUANG Jian,DU Qing-You,DING Yu and WANG Sheng-Qi 1591true in situ hybridization, immunohistochemistry, Western blot, Northern blot, cDNA expression microarray, co-immunoprecipitation and the tumorigenicity assay in vivo and in vitro etc. AKR1C2 gene expression and its effects was analyzed, including 68 pairs of HCC specimens and its adjacent para-cancerous tissues, 8 cases of normal liver tissues and QGY7703 cell line. Results showed AKR1C2 expression was up-regulated among these patients, when compared with those of para-cancerous and normal liver tissues. Over- expression of AKR1C2 is also found to be correlated with high metastasis potentiality of HCC. AKR1C2 overexpression stimulates DNA synthesis, apoptosis, growth in soft agar and promote tumor formation and lead to expression differences of tumor genes. AKR1C2 mediated the NF-κB-dependent resistance of QGY7703 cells to anti-fas killing. Intracellar binding of AKR1C2 and Cdk4 was found. Abnormal expression of AKR1C2 gene may contribute to the occurrence, advancement and invasiveness of HCC from Qidong, China, a liver cancer high risk area.]]> LU Dong-Dong,ZHANG Xi-Ran and CAO Xiang-Rong 1590true in silica. In order to investigate the level of alternative splicing and to search for novel splice variants, total 213 ESTs derived from 29 different tissues were collected and aligned against the genome sequence of the gene. 17 ESTs were found having different splicing type that could be classified into three types of alternative splicing. 14 of the ESTs were detected to have exon skipping, 2 intron not spliced and 1 splice site shift. The data suggested that exon skipping was possibly one of the major mechanisms of transcription regulation of HPCL2 gene.]]> NIU Yu-Xin,HU Song-Nian and YANG Huan-Ming 1589true 2O2-induced apoptosis of C2C12 myogenic cells, Hoechst 33258 staining was used to examine cell morphological changes and to calculate percentage of apoptotic nuclei. DNA ladder pattern on agarose gel electrophoresis was used to observe DNA fragmentation. The release of Smac/DIABLO from mitochondria to cytoplasm was observed by Western blotting. Activities of Caspase-3, Caspase-9 were assayed with Caspase Colorimetric Assay Kit and Western blotting. Full length Smac/DIABLO gene was transiently transfected in C2C12 myogenic cells by lipofectamine and then protein levels of Smac/DIABLO were analysed by Western blotting. The results showed that: (1)H2O2 (0.5 mmol/L) resulted in a marked release of Smac/DIABLO from mitochondrial to cytoplasm 1 h after treatment, activation of Caspase-3, Caspase-9 4 h after treatment and specific morphological changes of apoptosis 24 h after treatment; (2)Smac/DIABLO overexpression significantly enhanced H2O2 induced apoptosis in C2C12 myogenic cells as shown by specific DNA ladder pattern in agarose gel electrophoresis, increase of percentage of apoptotic nuclei and marked activation of Caspase-3, Caspase-9. These data suggested that H2O2 could result in apoptosis of C2C12 myogenic cells，and that release of Smac/DIABLO from mitochondrial to cytoplasm and the subsequent activation of Caspase-9 and Caspase-3 played important roles in H2O2-induced apoptosis in C2C12 myogenic cells.]]> JIANG Bi-Mei,XIAO Wei-Min,SHI Yong-Zhong,LIU Mei-Dong,TANG Dao-Lin and XIAO Xian-Zhong 1588true M.musculus protein BAB32214. Bioinformatics analysis suggested that the protein may be a nucleus protein regulating gene transcription. The new gene is comprised of four exons, with three intervening introns and it is localized to chromosome 5q35. It have been found that there are two CpG islands in 5′ UTR of this gene.]]> LI Feng,JIANG Wei-Hong,YIN Zhi-Hua,YANG Xu-Yu,FENG Xiang-Ling,LIU Wei-Dong and YAO Kai-Tai 1587true P＞0.05). It can be concluded that protein-chip reagent for detection of different HCV antibodies has higher sensitivity and specific than ELISA method and has the same accuracy as imported RIBA reagent. It will be a convenient and low costs reagent for diagnosis of HCV.]]> ZHANG Wen,ZHOU Mei-Fen,CHEN Li-Yan,DING Ya-Ping,CAO Heng-Jie,HUANG Jian,GENG Yong-Yao and WANG Sheng-Qi 1586true TANG Chao-Ke,HE Xiu-Sheng,YI Guang-Hui,WANG Zuo,YUAN Zhong-Hua,LIU LU-Shan,WEI Dang-Heng,WANG Yan and YANG Yong-Zong 1585true ZHANG Jing,SHI Xiu-Fan and YANG Heng-Fen 1584true CHEN She,YIN Xiang-Lei,ZONG Hong-Liang,FAN Kai-Yi,HUANG Chuan-Xin,GU Jian-Xin and SHEN Zong-Hou 1583true PENG Cong,TAN Chen,ZHANG Qiu-Hong,WANG Xiao-Yan,ZHOU Ming,HUANG He,WANG Rong and LI Xiao-Ling 1582true Fox c2）in cardiovascular development, mice lacking Fox c2 locus were produced by targeted mutation and the developmental anomalies in the aortic arch were found. Mice homozygotes for the mutation(Fox c2－/－) died embryonically from E12.5 (embryo days, E) to term. Although some of the homozygous mutants were born with abnormalities of the aortic arch, all of them died within 24 h after birth. Fox c2－/－ homozygous mutants all displayed the Type B or Type C of interrupted aortic arch, which is the same as human congenital cardiovascular anomalies. Mice heterozygous for the mutation developed normally. In situ hybridization analysis on E10.5 embryos showed that Fox c2 mRNA expressed at the third, fourth and sixth arch arteries strongly. However, left fourth arch arteries disappeared at E12.5 gradually. These results suggest that the Fox c2 plays indispensable role in the remodeling of left fourth arch arteries during the formation of aortic arch.]]> FU Yan,DENG Wei-Guo,LI Yi-Yang,LI Yu-Lin and N.MIURA 1581true A.A.SABOURY,F.HOSSEINI-KISHANI,M.REZAEI-TAWIRANI and B.RANJBAR 1580true GUI Jun-Hao,ZHAO Jing,XU Yan-Ming,YU Cui-Juan,JIA Lin-Tao,WANG Cheng-Ji and YANG An-Gang 1579true Oryza sativa L.ssp.indica) by whole-genome shotgun strategy. Pulse field gel electrophoresis (PFGE) was utilized to purify rice genomic DNA, which could efficiently remove the organelles (chloroplast and mitochondrion) DNA and reduce the contamination ratio from 3% to 0.2%. At the same time, the rice DNAs yielded from yellow seedlings and green seedlings were compared, and the differences between HB method and NIB method in high molecular weight(HMW) DNA isolation were also studied. Finally, a set of methods for obtaining the whole and highly pure rice genomic DNAs were proposed, which included culturing rice yellow seedling; isolating, embedding and lysising rice nuclei, purifying and recovering rice genomic DNAs in low melting point (LMP) agarose gel by PFGE. Ultrasonic treatment on HMW DNAs in the melting LMP gel at 38℃ was reported at first time, it facilitated to obtain the desired DNA fragments for construction of shotgun library and gradient libraries.]]> YANG Ji-Liang,WANG Hui,WANG Qing-Hua,LI Jun,LIU Bin,JIN De-Min and WANG Bin 1578true in vivo ischemia and in vitro hypoxia. Recently, cell shrinkage is proposed as an early prerequisite to apoptotic events leading to cell death. To understand the mechanism underlying potassium channel-mediated neuronal apoptosis, the temporal changes in neuronal cell body volume and the involvement of potassium channel in the apoptotic volume decrease were examined in a model of staurosporine (STS)-induced apoptosis of cultured hippocampal neurons. Nonselective potassium channel blocker tetraethylammonium (TEA) or raising extracellular K+ concentration significantly prevented STS-induced neuronal cell death. A similar neuroprotection was also observed by treatment with the selective high-conductance calcium-activated potassium channel (BK) blockers iberiotoxin and paxilline. These results indicate that potassium channels, especially BK channels, contribute to STS-induced neuronal apoptosis. Moreover, STS induced an early cell body volume decrease and this cell shrinkage was completely blocked by TEA or high extracellular K+. It is suggested that potassium efflux may be involved in the apoptotic volume decrease, which is probably one of the mechanisms underlying mediation of neuronal apoptosis by potassium channel.]]> WANG Ying,CHEN Ming,LI Xiao-Ming,LI Jian-Guo,HU Ping,XIAO Zhong-Ju and GAO Tian-Ming 1577true 2+ in hippocampal CA1 neurons is elevated during ischemia and at early period following reperfusion. This Ca2+ overload has been suggested to be involved in ischemic brain damage. In normal CA1 neurons, the major mechanism allowing Ca2+ entry from the extracellular compartment is the opening of voltage-gated Ca2+ channels. The aim of the present study was to explore whether L-type calcium channel in hippocampal CA1 neurons changed at early period of reperfusion after ischemia. Transient forebrain ischemia in a duration of 15 min was induced by the use of the 4-vessel occlusion method in rats. Single L-type calcium currents were recorded in cell-attached patches of actually dissociated hippocampal CA1 neurons. After ischemia, average total patch current of L-type Ca2+ channels significantly increased in CA1 neurons when compared with that of control. This ischemia-induced enhancement in channel function was due to a higher channel open probability. Further analysis of single channel kinetics showed a prolonged open time and an increased opening frequency in postischemic channels. It is suggested that the functional enhancement in L-type calcium channels may partially account for the postischemic increase in intracellular Ca2+ concentration of CA1 neurons following ischemia.]]> LI Xiao-Ming,LI Jian-Guo,HU Ping,YANG Jian-Ming,WANG Ying,LI Xiao-Wen and GAO Tian-Ming 1576true 8, and the virus titre of Ad-uPAR(+) was 0.5×108. 95D cells were infected with Ad-uPAR(－) and Ad-uPAR(+) in different multiplicity of infection (MOI). Norther blot analysis could detected the expression of antisense and sense RNA for 500 bp fragment of uPAR gene. With the increase of MOI, Western blot analysis indicated that with AD-uPAR(－) infection the protein level of uPAR of 95D cells decreased, and modified Boyden's Chamber assay suggested that the invasive ability of 95D cells also decreased obviously. In 95D cells infected with Ad-uPAR(+), both the mRNA and protein level of uPAR did not decrease, and cells still had high invasive ability. The results indicated that adenovirus is an efficient vector for transferring antisense RNA for uPAR into cells, and antisense RNA for uPAR could obviously inhibit the invasive ability of 95D human lung cancer cells.]]> SUN Xing-Hui,TAN Li,LI Ping,ZHANG Yu-Qing,WANG Xia,HOU min,SONG Hou-Yan and ZHU Yun-Song 1575true Sulfolobus acidocaldarius can be regulated up by different growth medium, this difference is regulated at mRNA level. The GS was purified by DEAE-Sepharose and Sephacryl S-300 to homogeneity. The molecular mass was determined to be dodecameric protein (630 ku) composed of identical subunits of 53 ku. The optical pH of this enzyme is about 7.3, the optical temperature of both γ-glutamyl transferase activity and biosynthetic activity is 90℃, the Arrhenius plots show that the active energy is 47 kJ/(mol·K) for transferase activity and 29 kJ/(mol·K)(40～55℃),10 kJ/(mol·K)(55～90℃) for biosynthetic activity respectively. GS was stable at 78℃ in the presence of Mn2+, the Km values were 3.5 mmol/L, 1.3 mmol/L, 0.5 mmol/L and 0.24 mmol/L for hydroxylamine, glutamine, ADP and Mn2+ respectively. The inhibitors experiments showed that the catalytic activity of GS from Sulfolobus acidocaldarius unlike that of others was regulated solely by feed-back inhibition through L-alanine and glycine, the normal inhibitors such as L-tryptophan, L-histidine and 5′-AMP have no inhibitory effect on this enzyme. L-alanine and glycine have shown synergistic effect on catalytic activity of GS. The post-translation modification like other gram positive bcteria is not regulated by adenylylation/deadenylylation system.]]> YIN Zhi-Min,CHEN Qun-Ying,SIMA-Jian,WU Yi-Fan and ZHANG Shuang-Quan 1574true I was about 4.3 on IEF. The influence of pH and temperature on the activity of hk-1 was studied. The whole procedure is suitable for large-scale production.]]> WANG Ju,HONG An,SUN Fen-Yong and XIE Qiu-Ling 1573true ZHAO Tian-Fu,ZHANG Jian,LI Jing,CAO Yu-Jing,LI Su-Min,ZHOU Jia-Xi,LI Fu-Yang,YAO Li-Bo and DUAN En-Kui 1572true Eucommia antifungal protein (EAFP) crystals can be easily grown into big crystals in several hours. By in situ atomic force microscopy (AFM) the dynamic topographic changes were observed on the surfaces of several EAFP crystals and growth rates were measured at different supersaturations of the protein solution. The results of AFM experiments indicated that growth rates of EAFP crystals were strongly and directly related to the supersaturations, in addition to the inherent structural rigidity and the interior stability of the molecule. At higher supersaturation (σ=1.78) the EAFP crystals grew very fast; at moderate supersaturation (σ=1.5) the growth rates were 12 nm/s and 24.2 nm/s along the crystallographic axes b, c of the {100} surface respectively, which were faster than that of lysozyme (6～7 nm/s). Even at lower supersaturation the EAFP crystals grew almost as fast as other protein crystals did. The effects of the concentration of precipitator on crystal growth observed on the crystal growth of AFM at lower supersaturation were also presented.]]> WANG Sheng,XIANG Ye,LI Gen-Pei and WANG Da-Cheng 1571true I 4～8, altogether (905±57) and (881±32) spots were detected on the 2-DE map of normal and globozoospermic sperms, respectively. 607 spots were matched between the two groups. Mass spectrometry were performed with 16 proteins which were found to be absent in the round-headed sperms and 1 which expressed in a much lower concentration. Altogether 8 protein spots were identified and their possible roles are discussed. Among them, 3 spots were Golgi apparatus-related, 2 spots were subunits of proteasome and 2 spots were zinc finger proteins. Their absence or down-expression possibly interrupt spermatogenic process.]]> LUO Ke-Li,FAN Li-Qing,ZHU Wen-Bing and LU Guang-Xiu 1570true treY and treZ) encoding for trehalose-producing enzymes, a maltooligosyl trehalose synthase (MTSase) and a maltooligosyl trehalose trehalohydrolase (MTHase), had been cloned, expressed and sequenced from thermophilic archaebacterium Sulfolobus shibatae B12. The nucleotide sequences of treY and treZ indicated proteins with lengths of 728 and 559 amino acids and molecular masses of 86 ku and 65 ku, respectively. treY and treZ genes from S.shibatae B12 show high sequence homologies with other microbial MTSase and MTHase genes, such as S.solfataricus P2 (93% and 76% identity),S.solfataricus KM1(97% and 95% identity),Sulfolobus acidocaldarius ATCC 33909 (63% and 66% identity),Arthrobacter sp. Q36 (48% and 50% identity),Rhizobium sp. M-11(48% and 52% identity),Brevibacterium sp. (50% and 52% identity). Phylogenetic trees of these enzymes genes were constructed by analysis class of their overall nucleotide sequences. The amino acid sequences of MTSase and MTHase showed identity with members of the glycosyl hydrolase family 13 (α-amylase family). Four important regions highly conserved in the glycosyl hydrolase family 13 exist in the amino acid sequences of all these trehalose-producing enzymes. It is suggested that MTSase and MTHase are members of glycosyl hydrolase family 13 and treY and treZ maybe derived from a common original α-amylase gene.]]> WU Jin,YU Wei-Ting,WANG Hui,LIU Li,WANG Shao-Xiao and ZHANG Shu-Zheng 1569true GUO Dong-Sheng,ZHU Cheng-Gang,ZHANG Yao-Zhou and WU Xiang-Fu 1568true CUI Lin-Yang,JIANG Nan and CHEN Shi-Jun 1567true LI Wen-Yong,Qi Bing,WANG Yu-Ge,YU Wei-Dong,DU Miao and CHEN Qing-Xuan 1566true ZHU Zi-Yu,WEN Duan-Cheng,HAN Zhi-Ming,LEI Lei,LIU Zhong-Hua,KOU Zhao-Hui,XU Ying,WANG Min-Kang and CHEN Da-Yuan 1565true YUAN Zhong-Hua,YANG Yong-Zong,YIN Wei-Dong,YI Guang-Hui and WU Meng-Jin 1564true MTHFR gene influences the enzyme activity and is correlated with susceptibility to several tumour types. In order to compare the association of the MTHFR C677T polymorphism with susceptibility to esophageal squamous cell carcinoma (ESCC) in a Northern Chinese population, high-speed real-time PCR and melting curve analysis were used. The MTHFR C677T genotypes were determined in 189 patients with ESCC and 141 unrelated healthy controls. The results showed that the MTHFR C677T C/C, C/T and T/T genotype frequencies among healthy controls were 17.7%, 38.3%, and 44.0%, respectively. There was no significant difference in MTHFR T/T genotype frequency between ESCC patients (42.3%) and healthy controls (χ2=0.089, P＞0.05) whereas the C/T genotype frequency among ESCC patient (49.2%) was only slightly higher than that among healthy controls (χ2＝3.890, P＜0.05). However, the frequency of the C/C genotype among ESCC patients (8.5%) was significantly lower than that among healthy controls (17.7%) (χ2=6.37, P＜0.05). The C/C genotype significantly reduced the risk for developing ESCC compared to the combination of the C/T and T/T genotypes (OR=0.43, 95% CI=0.22～0.84). The reduced risk was more evident among smokers and patients with family history of upper gastrointestinal cancers. It can be concluded that the MTHFR C677T homozygous wild type may play a protective role in the ESCC development in the Northern Chinese population.]]> ZHANG Jian-Hui,LI Yan,GUO Wei,WANG Rui,M.SARBIA,S.KIEL,WEN Deng-Gui,WEI Li-Zhen,CHEN Zhi-Feng,HE Ming,KUANG Gang,ZHANG Li-Wei,WU Ming-Li and WANG Shi-Jie 1563true TAN Yun-Nian,TAO Yong-Guang,SONG Xin,TANG Min,AI Mi-Dan and CAO Ya 1562true Arabidopsis thaliana PAP-specific phosphatase was predicted by use of various existing methods on sequence comparison, secondary structure prediction, three dimensional structure prediction and simulation. It was a structure similar with that of Hal2p in Saccharomyces cerevisiae, consisting of an α＋β N-terminal domain and an α/β C-terminal domain. In the predicted structure, possible binding sites for Mg2+, as well as for other metal ions, and the structural base sensitive to Na+ were found. These sites were related with the biochemical function of Arabidopsis thaliana PAP-specific phosphatase. The structural and functional analysis suggested that the theoretical structure of Arabidopsis thaliana PAP-specific phosphatase, having been deposited in PDB, is not reasonable.]]> Peng Yi and Zeng Zong-Hao 1561true ZENG Liang,LI Min,SONG Xin,TAO Yong-Guang,TANG Min,LI Wei and CAO Ya 1560true HU Zhi,TAO Yong-Guang,TANG Fa-Qing,YANG Li-Fang,ZHAO Yan,ZENG Liang,LUO Fei-Jun and CAO Ya 1559true JIANG Dai-Feng,YING Wan-Tao,WAN Jing-Hong,QIU Zong-Yin,QIAN Xiao-Hong and HE Fu-Chu 1558true 125I-apoCⅢ to HepG2 cells revealed a saturable binding to HepG2 cells with a Kd of （9.53±1.03）×10-9 mol/L. The maximum specific binding capacity (Bmax) was (3.28±0.31) μg/g. The results showed that there were specific, saturable, and reversible binding site of apoCⅢ on HepG2 cells, and it was identified that HepG2 cell line was a useful model for the study of human hepatic apoCⅢ receptor. In the following, the effects of insulin and glucagon on apoCⅢ receptr of HepG2 cells were examined respectively. The results showed that insulin had no effect on the Kd value, but significantly increased the Bmax value of apoCⅢ receptor; and glucagon had no effect on the Bmax, but decreased the affinity constant of apoCⅢ receptor. These results indicated that human hepatic apoCⅢ receptor was possibliy under the different regulations of insulin and glucagon.]]> LIU Hao,LIU Bing-Wen and LI Qiang 1557true E.coli DH5α；The binding between M3M4 and MAB363 could be competitively inhibited by one of solid-synthesized epitope peptides: RLRNPSKD. There were identical sequences among them: NPS. Deleted with NPS, the peptide could not inhibit the binding of MAB363 to M3-M4. These results demonstrate that NPS in M3-M4 loop is the B cell epitope recognized by MAB363，which may be important for developing a practical immunization strategy against excitotoxic brain injury.]]> KANG Xiao-Nan,SUN Chang-Kai,FAN Ming,XUE Yan-Ning,SHAO Ning-Sheng,ZHAO Jie,HAN Da-Yue and SHI Guang-Xia 1556true LI Peng-Cheng,CHEN Shang-Bin,LUO Wei-Hua and LUO Qing-Ming 1555true 121 was constructed. After transfection, individual viral plaques were isolated and amplified in HEK293 cells. Confirmed by PCR, both Adeno-X-VEGF121 and Adeno-X-LacZ were propagated in HEK293 cells and were purified by CsCl density gradient centrifugation. Adenovirus-mediated VEGF121 gene transfer promotes ECV304 proliferation and formation of capillary-like structures in vitro. The expression of VEGF121 by adenovirus-infected ECV304 was quantified by enzymelinked immunosorbent assay. Peak VEGF121 production was achived at 7～10 days after infection. The conditioned media from ECV304 infected with Adeno-X-VEGF121 markedly enhanced vascular permeability. These data may support that adenovirus-mediated VEGF121 cDNA gene transfer could provide a useful strategy for efficient delivery of VEGF121 in the treatment of ischemic diseases.]]> WANG Yue-Xiang,YANG Jian,MA Chun-Gu,YANG Xin-Ying and SONG Hou-Yan 1554true CHEN Xiang-Gui,LI Yong,ZHAO Ru-Bing,PEI Xin-Rong,ZANG Ming-Xi,GAO Li-Fang and CHEN Jun 1553true QIAN Wei,LIU Fei,ZHU Li,GONG Cheng-Xin and JIN Shu-Yi 1552true Pichia pastoris expression vector pPICZαA, the resulted recombinant expression vector transformed SMD1168 competent cells and obtained engineering Pichia pastoris strain SMD1168/pPICZαA-FL. After high concentration ZeocinTM screening, phenotype identification, inductive expression, SDS-PAGE and Western blot analysis of culture supernatant, an engineering Pichia pastoris strain SMD1168/pPICZαA-FL-7 in which gE gene fragment deleted signal peptide was expressed in high levels was obtained. SDS-PAGE and Western blot indicated that expression product of gE fragment deleted signal peptide in culture supernatant of SMD1168/pPICZαA-FL-7 was about 80 ku, a little larger than expected. Gel scanning and Bradford protein analysis results showed that expression product reached 11.7 g/L, or 13.49% of total culture supernatant protein in SMD1168/ pPICZαA-FL-7.]]> AO Jing-Qun,WANG Jin-Wen,Chen Xin-Hua,WANG Xun-Zhang,LONG Qing-Xin and LOU Gao-Ming 1551true 1, P5-Di（adenosine-5′）pentaphosphate］ during this process was investigatived. The results revealed that: 1) TRAIL-induced apoptosis is both dosage- and time- dependent, which correlates with the remarkable increase of ［Ca2+］ in Novikoff cells; 2) Ap5A retards the TRAIL-induced apoptosis as well as down-regulates ［Ca2+］ in Novikoff cells. These observations indicate that the mechanism of TRAIL-induced apoptosis and Ap5A retards it through down and up-regulating intracellular calcium concentration respectively.]]> LAI Qiu-An,LIU Yan-Xin,HAO Xue-Mei,TANG Ai-Hui,Wei-Wei,WANG Shi-Qiang and Sun Jiu-Rong 1550true ZHOU Ji-Yong,CHEN Ji-Gang,WANG Jin-Yong and JIMMY KWANG 1549true YUAN Can,Lü Qing-Lan,CHEN Guang-Wen,LIU Ying,WANG Yao-Ling,LIU Hai-Jun,ZOU Jiang and XIAO Xian-Zhong 1548true P＜0.05) to the level closing to the low metastatic hepatocarcinoma cells. The level of galactocerebroside which was the precursor of sulfated lactosyl ceramide did not change significantly. This indicated that the sulfation was inhibited after retinoic acid treatment. Therefore the cerebroside sulfotransferase gene were transferred into hepatocarcinoma cells and selected sulfated lactosyl ceramide-highly-expressed clones. Through adhesive assay, it was observed that the control cells were quite poor in adhesive to vitronectin and laminin, the transfected cells were significantly enhanced in the adhesion to vitronectin and laminin. After 45 minute incubation, the adhesive percentage of the cells was 3～4 time higher in CST-transfected cells than those in control cells. By animal experiments there were more metastatic foci in the liver or lymph nodes in CST-transfected groups than the control (P＜0.05). The results suggested that the high expression of SM3 in hepatocarcinoma cells promoted metastasis in nude mice and enhanced the adhesion to vitronectin.]]> WU Xing-Zhong,LI Wei,BEN Jia and ZHA Xi-Liang 1547true in vivo. Genotype of coding region single nucleotide polymorphisms (cSNPs) in NOR1 gene were performed by sequencing in 144 unrelated NPC subjects and 144 control subjects which matched in age, sex and residence. Association analysis suggests that there is linkage disequilibrium between the 2 cSNPs, both of them associated with NPC. And the relative risk of 2 cSNPs and their haplotypes were 1.36, 1.64 and 1.37 respectively. Both of the two SNPs could change the sequence of NOR1 protein, which might influence it's structure and function. The results indicated that the SNPs in NOR1 gene may play a certain role in carcinogenesis and development of NPC.]]> XIONG Wei,ZENG Zhao-Yang,XIAO Bing-Yi,XIONG Fang,NIE Xin-Min,FAN Song-Qing,PENG Cong,LI Wei-Fang,WANG Rong,SHEN Shou-Rong,LI Xiao-Ling and LI Gui-Yuan 1546true ZHENG Jing-Min,LIU Zhi-Hong,ZHANG Xin and LI Lei-Shi 1545true ZHANG Feng-Yue,YIN Zhi-Xiang and XU Jin 1544true V, the diffusion constant D and the randomness parameter r of the steady state are all be obtained. By comparing the simulated curve with experiment about drift velocity V, diffusion constant D and randomness parameter r versus ［ATP］ and external load F, the kinetic behaviors of a molecular motor under external load F are qualitatively analyzed.]]> ZHAN Yong,WU Wei-Xia,ZHAO Tong-Jun,GUAN Rong-Hua and MEI Jun-Ping 1543true H.pylori (Hp) adhesin conservation region(AB) in vitro so that to determine the feasibility of AB in Hp vaccination. ELISA assay was used to measure AB-specific antibody in serum of Hp infected patients. The proliferation of T cell in response to AB was examined by MTT test. Flow cytometry was used to evaluate the increase in FasL expression on T cells under the stimulation of Hp AB,T cell apoptosis induced by AB was detected by DAP assay.The effect of anti-AB serum on Hp binding of human gastric carcinoma cell lines was determined by light microscopy. Antibodies were detected in sera samples from 55 patients by ELISA, with RUT as parallel compare. Kappa value is 0.76. The low dose AB was capable of stimulating proliferation of T cells from Hp positive subjects. The effect of AB was significantly lower in both the induction of apoptosis and FasL expression of T cells than that of ATCC26695. Anti-AB serum could partially inhibit Hp binding to gastric epithelial cells. Under light microscopy, the adhesion of Hp to MGC-803 was significantly inhibited when pretreated the bacteria with anti-AB rabbit serum, compared with negative control which pretreated with pre-immunized rabbit serum. AB was a safe, immunogenicity thallus component, which can stimulate humoral and cellular immunity.Its antibody was capable of preventing the binding of Hp to gastric epithelial cell.]]> BAI Yang,ZHANG Ya-Li,CHEN Ye,WANG Ji-De and ZHOU Dian-Yuan 1542true ZHENG Bin,WEN Jin-Kun,HAN Mei and ZHOU Ai-Ru 1541true Km, vmax and Ea of HLADH-catalyzed asymmetric reduction of acetyltrimethylsilane were 2.67 mmol/L, 0.118 mmol/(L·min·mg) and 37 kJ/mol respectively, while the corresponding parameters for its carbon analogue were 3.56 mmol/L, 0.084 mmol/(L·min·mg) and 61 kJ/mol.]]> LOU Wen-Yong,ZONG Min-Hua,WANG Ju-Fang and WU Hong 1540true WU Heng,SHEN Min-Xiong,LIANG Yu-Long,DUAN Ling-Ling,WANG Li-Ying,XU Zhen and ZHA Xi-Liang 1539true Musca domestic pupae was isolated. The purification procedure entailed extraction with aqueous buffer, GRBC (glutaraldehydeized red blood cells) adsorption, affinity chromatography on Sepharose-4B and gel-filtration on SephadexG-200. By SDS-PAGE, purified MPL yielded three bands, with a total molecular mass of 84 ku. By gel-filtration, the molecular mass of MPL was determined to be 86 ku. MPL exhibits high affinity towards D-galactose. The hemagglutination activity of MPL was independent of Ca2+, heat liable and stable in the range of pH 6～9. Antibacterial activity of MPL has been observed against Escherichia coli, Bacillus subtilis and Salmonella typhi.]]> CAO Xiao-Hong,CHEN Yi,ZHANG Yan and CHEN Jin-Ying 1538true E.coliM15, the recombinant proteins were expressed successfully when induced with IPTG. The expressed recombinant protein accounted for more than 50% of total bacterial proteins. The expressed products formed inclusion body in E.coli. After extracted from bacterial cells and washed, it was dissolved in solution containing 8 mol/L urea and then purified by using immobilized metal ion affinity chromatography (IMAC) effectively with a purity of over 95%. Then the recombinant protein was renatured after the denaturants were removed gradually by dialysis. The protein was identified by the determination of its N-terminal amino acid sequence, molecular mass, isoelectric point etc. The results indicated that the primary structure of the expressed protein accorded with the theoretics. Endothelial cell proliferation assay, endothelial cell migration assay and chick chorioallantoic member assay, the bioactivity of vasostatin was investigated. It was proved that vasostatin can inhibit endothelial cell proliferation and migration, and inhibit angiogenesis of chick chorioallantoic member.]]> LI Yong-Hong,Wang Jun-Zhi,Rao Chun-Ming and Zhang Yi 1537true Eisenia fetida extract (mainly a group of anti-tumor protein components) by hydrophobic interaction chromatography and ion exchange chromatography. The molecular mass assayed by SDS-PAGE and isoelectric point of ARSP1 were 28 ku and less than 3.8, separately. However, several coterminous bands could be observed by PAGE of natural ARSP1 and several coterminous peaks of ARSP1 were also detected with MALDI-TOF-MS when the relative molecular mass of three main peaks are 24 645, 25 052 and 25 281, separately. The N-term amino acid sequence of ARSP1 was assayed as following: I(V)IGGT(S)N(D)ASPGEFPWQLSQTRGGSHS. And, a result that ARSP1 is highly homologous with serine proteases was concluded by the comparison of N-term amino acid sequences. In vitro, the cytotoxicity of ARSP1 was not only identified by phase-contrast microscopy observation of apoptotic cells, but also studied further by the localization of fluorescent antibodies. By Schiff's staining, ARSP1 was identified to be glycoprotein or glycopeptide. By fibrin plate assay, ARSP1 was identified to be a plasmin and also a plasminogen activator, and the fibrinolytic activity was inhibited by PMSF (an inhibitor of serine proteases).]]> XIE Jiang-Bi,GUO Zhen-Quan,WENG Ning,WANG Hong-Tao,JIANG Guan-Qun and RU Bing-Gen 1536true ZHANG Hui,ZHAN Jin-Biao,XU Lin-Hai,YAN Zhi-Kun and WANG Ke-Yi 1535true P＜0.05) respectively. But statistically, there was no difference in the levels of STAT3 and phospho-STAT3 among the three groups (P＞0.05). All the results suggest that there is a possible involvement of Ras/ P44/42 MAPK pathways in the cancerization process of leukemia cell, while JAK/STAT3 pathway makes no contribution to the process of radiation carcinogenesis.]]> FU Zhi-Chao,CAI Jian-Ming,HAN Ling,WANG Feng-Mei,HUANG Ding-De,HUANG Yue-Cheng,LI Bai-Long and GAO Jian-Guo 1534true PTEN in HEK293 cells could lead to apoptosis and cell cycle arrest. The wild-type and mutant T910G of PTEN expression plasmids were constructed and transfected into PTEN-null HEK293 cells respectively. Apoptosis was evaluated by the appearance of cytosolic low molecular DNA ladder on the gel. Cell cycle was determined by flow-cytometric analysis. The Western blot analysis was performed to determine the phosphorylation levels of PKB/Akt and MAPK. The present data showed that the overexpression of PTEN in HEK293 cells could induce apoptosis and resulted in an increase in G1 cell population through inhibiting PKB/Akt and MAPK phosphrylation stimulated by PDGF. Mutant PTEN cause less apoptosis and G1 arrest than wild-type PTEN. MAPK dephosphorylation caused by mutant PTEN was not so significant as by wild-type PTEN. These data suggested that PTEN may exert its tumor-suppressive effects through both the inhibition of cell cycle progression and the induction of apoptosis.]]> ZHANG Li-Neng,Yu Qiang and ZHA Xi-Liang 1533true P＜0.01). Optical properties of a certain of tissue at 532 nm and 808 nm laser and their linearly polarized laser irradiation had either prominent distinction (P＜0.01). Optical properties of human normal bladder tissue at a certain wavelength of laser and its linearly polarized laser had obvious distinction. And optical properties of human bladder cancer tissue at a certain wavelength of laser and its linearly polarized laser irradiation had not prominent distinction. Attenuation of all of the forward scattered photon fluxes i(x), the backward scattered photon fluxes j(x), the total scattered photon fluxes I(x) of human bladder cancer tissue at 532 nm and 808 nm laser and their linearly polarized laser irradiation is obviously bigger than one of human normal bladder tissue. And that light intensities of their forward scattered photon fluxes i(x) was obviously bigger than one of their backward scattered photon fluxes j(x).]]> WEI Hua-Jiang,XING Da,WU Guo-Yong,GU Huai-Min and JIN Ying 1532true GUO Wei-Sheng and LUO Liao-Fu 1531true Arabidopsis thaliana was made, and a direct comparison of genctic code preference among Arabidopsis thaliana, Homo sapiens and Escherichia coli was carried out.The results show that Arabidopsis thaliana like Homo sapiens its frequency of codon usage is obviously different from Escherichia coli, and there is a considerable difference between Arabidopsis thaliana and Homo sapiens. The data will give some suggestions to those researchers who want to introduce a animal gene into plant or a plant gene into bacteria.]]> FAN San-Hong,GUO Ai-Guang,SHAN Li-Wei and HU Xiao-Ping 1530true Cuscuta chinensis Lam., one of the most important traditional Chinese medicines for tonifying liver and kidney, enhances mitogen-activated protein kinases (MAPKs) activity and as a consequence induces neurite outgrowth in PC12 cells. The effect of Cuscuta chinensis Lam. extract on neurite outgrowth and its potentriated activation of MAPK were similar to that of nerve growth factor. PD98059, a specific inhibitor of MAPK, inhibited the effect of the extract on the phosphorylation of MAPK, suggesting the extract induce the PC12 cells differentiation linked to the MAPK cascade. Furthermore, the extract prevents apoptosis of PC12 cells caused by serum deprivation, indicating that it has neurotrophic factor-like activity.]]> LIU Jian-Hui,JIANG Bo,BAO Yong-Ming and AN Li-Jia 1529true Saccharomyces cerevisiae (yeast) genes and their introns, a remarkable phenomenon was discovered that generally the introns of the genes with higher transcription frequencies are longer, and the introns of the genes with lower transcription frequencies are shorter. This suggests that the longer introns of genes with higher transcription frequencies may contain some characteristic sequence structures, which could enhance the transcription of genes. Therefore, two sets of introns of yeast genes were chosen for further study. The transcription frequencies of the first set of genes are higher (＞30), and those of the second set of genes are lower (≤10). Some oligonucleotides are detected by statistically comparative analyses of the occurrence frequencies of oligonucleotides (mainly tetranucleotides and pentanucleotides), whose occurrence frequencies in the first set of introns are significantly higher than those in the second set of introns, and are also significantly higher than those in the exons flanking the introns of the first set. Some of these extracted oligonucleotides are the same as the regulatory elements of transcription revealed by experimental analyses. Besides, the distributions of these extracted oligonucleotides in the two sets of introns and the exons show that the sequence structures of the first set of introns are favorable for transcription of genes.]]> ZHANG Jing and SHI Xiu-Fan 1528true in vivo valproic acid administration on ERK-1/2 signaling pathway, and to elucidate the molecular mechanisms underlying the therapeutic effects of valproic acid, male Wistar rats were divided into two groups (each 20 animals), one group was treated with valproic acid chow (3.6 g/kg) for 4 weeks, the other group had access to normal chow as control. By the end of the 4th week, rat brains were removed immediately on decapitation and dissected on ice, and the hippocampus and frontal cortex were obtained. The total proteins or nuclear proteins of rat brain hippocampus and frontal cortex were prepared. Levels of the phosphorylated, active forms of MEK,ERK-1/2,RSK1,CREB and expression levels of Bcl-2 were assayed by Western blot analysis. DNA binding activity of transcriptional factor AP1 was determined by EMSA. Valproic acid increased the activities of MEK,ERK-1/2,RSK1,CREB and AP1, and up-regulated the expression of Bcl-2 in rat brain hippocampus and frontal cortex. These data suggest chronic treatment of valproic acid activates ERKs signaling pathway and up-regulates the expression of Bcl-2 proteins in central nerve system(CNS), which may associate with the therapeutic efficacy of valproic acid in the treatment of manic-depressive illness.]]> LI Jian-Ling,TANG Cen-E,CHEN Zhu-Chu and XIAO Zhi-Qiang 1527true EcoRⅠ～BamHⅠ sites of pYD1. The recombinant plasmids pYD1-Trunc4/Trunc16/Trunc20/Trunc32 were transformed into yeast EBY100. As control, pYD1-mdb3 was also transformed. After four groups fusion protein were expressed after adding galactose, the Cl- transport activity was measured by using a fluorescent probe SPQ. The result demonstrated that the transport activity of band 3 was decreased when knockout Lys892～Val911 of AE1-C-terminal domain, but the transport activity didn't decrease further when knockout Asn880～Val911 of AE1-C-terminal domain. These results showed that Lys892～Phe895 amino acids influenced the anion transport of band 3 transmembrane domain.]]> LIU Li-Mei,FU Guo-Hui,WANG Tian-Ying,JIANG Xiao-Shu,GUO Zhuo-Wei and SHI Cong-Ning 1526true Arabidopsis thaliana including npq1 (lutein-replete and violaxanthin deepoxidase-deficient), lut2 (lutein-deficient) and lut2-npq1 (double mutant) under high light condition was characterized. There was no obvious difference in ratio of Chl a/b between mutants and wildtype, while Fv/Fm in mutants increased to some extents. The total xanthophyll pool (V+A+Z) increased significantly in lutein-deficient mutants (lut2 and lut2-npq1). The NPQ induction by high light was markedly inhibited in lut2-npq1 and npq1, but showed less inhibition in lut2. The trend of NPQ value in mutants and WT exposed to PPFD during high light illumination of 2 000 μmol·m-2·s-1 for 1～9 min was WT＞lut2＞npq1＞lut2-npq1. qP in all three mutants decreased in comparison with wildtype. Three xanthophyll-dificient mutants exhibited less resistance to photoinhibition than the WT. The sequence of tolerance to photoinhibition was the same as the changes in NPQ among WT and mutants (WT＞lut2＞npq1＞lut2-npq1). The results indicated that xanthophyll cycle was related not only directly to NPQ formation, but also to qP.]]> PENG Chang-Lian and LIN Gui-Zhu 1525true YUAN Hong-Feng,WANG Dong-Mei,LI Hai-Min,CHEN Lin,WANG Xiao,BAI Ci-Xian,YUE Wen and PEI Xue-Tao 1524true ZHANG Bi-Cheng,ZHOU Ming,ZHOU Hou-De,XIAO Bing-Yi,NIE Xin-Min,ZHU Shi-Guo,LI Wei-Fang,LI Xiao-Ling and LI Gui-Yuan 1523true Ito) of isolated rabbit atrial myocytes，standard whole-cell patch clamp technique was used to record Ito before and after administration of extracellular BmkTXKβ with multiple concentrations. The results showed that: (1) At a dose of 1 μmol/L, it decreased Ito by 41.4% (n=16, P＜0.001) at membrane potential of +50 mV ［from (13.63±0.87) pA/pF to (7.98±0.78) pA/pF］. After washout, Ito restored to (11.18±0.82) pA/pF (n=6, P＜0.01). (2) It significantly reduced Ito in a clearly concentration-dependent manner at the range of 0.01～100μmol/L with an IC50 value of 0.95 μmol/L (n=10, P＜0.01), but without any change in frequency-dependence (n=6，P＞0.05). (3) In the absence and presence of BmkTXKβ (1 μmol/L),the activation curves from relative conductance almost overlapped, whereas steady-state inactivation curve shifted to left from （－23.6±2.7） mV to （－35.3±3.6） mV at V1/2 point significantly （n=8，P＜0.05）. The time for 50% recovery delayed obviously from（51.2±8.5） ms to （93.5±13.4） ms in the absence and presence of 1 μmol/L BmkTXKβ（n=9，P＜0.01）. The results show that BmkTXKβ exerts direct blocking effect on Ito in rabbit atrial myocytes, which is mainly caused by a strong suppression effect on inactivation duration and prolongation of the recovery duration from inavtivation.]]> HU Dan,HUANG Cong-Xin,JIANG Hong,LI Geng-Shan,CAO Zhi-Jian,LI Wen-Xin and WANG Shi-Min 1522true Pseudomonas exotoxin A (PE)-derived peptide was fused to N-terminus of r-casp3. The expression of the interest genes was detected upon induction with ponasterone. The genes of r-casp3 and cr-casp3 were demonstrated to effectively cause cell death by MTT assay and cell counting. Cells that expressed r-casp3 or cr-casp3, but not wt-casp3, underwent apoptosis in a comparable level as determined by cell cycle analysis, genomic DNA ladders, and electronic microscopy. These results prove that unlike wild-type caspase-3 which is inactive unless proteolytically processed by upstream caspase, both recombinant caspase-3s are naturally active, and the N-terminal fusion of PE translocation domain does not interfere with the natural caspase-3 activity, suggesting their applications on the construction of novel tumor therapeutics that efficiently translocate to the cytosol of tumor cells and cause cell death.]]> JIA Lin-Tao,ZHANG Li-Hong,YU Cui-Juan,JI Zong-Ling,CAO Yun-Xin,WANG Cheng-Ji and YANG An-Gang 1521true Escherichia coli (ETEC) is a major pathogen that evokes acute diarrhea among children worldwide and travelers to developing countries. However, there is no ideal vaccine against it yet. In an effort to develop a subunit vaccine for ETEC, a translational fusion with cholera toxin B subunit (CTB) upstream of CS3 was constructed. The fusion protein synthesized in E.coli had a molecular mass of 29 ku, as expected and retained the antigenicity of both CTB and CS3 as confirmed by Western blot analysis with the polyclonal anti-CTX rabbit serum and the monoclonal anti-CS3 mouse serum, respectively. The 6×His-tagged CTB/CS3 protein was purified by Ni-NTA affinity chromatography followed by renaturation. A fraction of the fusion protein could form pentamers and these pentamers retained the ability to bind GM1-ganglioside. Mice immunized by intraperitoneal injection with the fusion protein produced anti-CTB and anti-CS3 serum IgG and secretory IgA. Furthermore, it was shown that fusion to CTB increased the systemic and mucosal immune responses against CS3 to some extent.]]> MI Kai-Xia,LI Ji,ZHANG Zhao-Shan and FANG Rong-Xiang 1520true 32P, respectively. By using nucleic acid spot hybridization (NASH), relative RNA loading (RRL) of both gemonic and satellite RNAs were quantitatively determined from the systemically infected hosts tissue. At 16～20℃, the radish-derived CMV-R3 containing no satRNA was inoculated on four host plants and examined 15 days, 30 days, and 75 days post inoculation, respectively. The resulting RRL showed a declining trend during the test period. On day 15, the RRL for genomic RNA differed obviously among the hosts in the order of Nicotiana.tobacum＞N.glutinosa＞N.clevelandii＞tomato. Meanwhile, the RRL for the CMV-RS, another radish isolate containing high copies of satRNA, was examined 5 days and 15 days after inoculation on the same hosts with CMV-R3 being included as a non-satRNA control. The RRLs for both genomic RNA and satellite RNA of the CMV-RS displayed a similar host- and time-effect trend. On all the inoculated hosts, the RRL increased from day 5 to day 15 and RRL of CMV-RS for both genomic RNA and satRNA was in the quantitative order of N.tobacum＞N.glutinosa＞Nicandra.physalodesand tomato. At 18～21℃, CMV-HC4, a severe tomato isolate containing a necrosis satRNA, was tested after 5 days, 10 days, and 15 days inoculation on 5 hosts. The RRLs of HC4 genomic and satellite RNAs were under the influence of host and inoculation time. The RRLs for satRNA and genomic RNA were similar but had some degree differences among the hosts. On day 10 post inoculation, the relative amount of both genomic RNA and satRNA was ordered as tomato＞N.glutinosa＞N.tobacum. The results also showed that different CMV isolates have obvious preference among hosts for replication and accumulation of viral RNAs.]]> CHEN Ji-Shuang,CHAI Li-Hong and WU Ping 1519true KANG Xiao-Nan,SUN Chang-Kai,FAN Ming,SONG Zhan-Jun,ZHAO Jie,WANG Ji-Qing and SHI Guang-Xia 1518true +mRNA was purified from total RNA by Oligotex mRNA Kit. cDNA was synthesized using SuperScriptTM Choice System For cDNA Synthesis Kit. The quality of synthesized cDNA was detected by chemical light method using PolyA+ mRNA probe. The double-strand cDNA was ligated into the EcoRⅠ site of pGADT7 vector and the cDNA library from the esophageal cancer cell line for yeast-hybrid system was constructed. The titer of the amplified cDNA library was 1.19×109 cfu/ml，The inserted fragment size of recombinants was from 0.5 kb to 6.0 kb and the percentage of recombinant clones was about 50％. NF-κB element binding factors were screened from this cDNA library by yeast one-hybrid technic and more than 360 clones were obtained on the SD/-his/-leu/［＋15 mmol/L 3-AT］. Plasmids of 91 clones which were larger than 2 mm in diameter were isolated from yeast and transformed to E.coli 30 positive recombinants had been obtained since plasmids were extracted from E.coli and digested by EcoRⅠ and validated by yeast one-hybrid assay, 9 clones of them were sequenced and the sequence was compared with GenBank/BLAST database. Results showed that protein product of some genes obviously conformed to consensus sequence of NF-κB element binding domain of p65 or p50 in the key sites of amino acid residues. These results determined that the human esophageal cancer cDNA library for yeast-hybrid system had a higher quality.]]> XU Li-Yan,LI En-Min,NIU Yong-Dong,CAI Wei-Jia,HAN Ming,WU Bing-Li,ZHANG Can,SHEN Zhong-Ying and ZENG Yi 1517true in vitro, and the expressed hCGβ protein was mainly with intracellular state. With 20 μg PCR3.1-hCGβ plasmid DNA through intramuscular inoculation after bupivacaine-HCl inducing, the immunized BALB/c mice could express hCGβ antigen to induce both intensive hCGβ-specific humoral immune responses and CTL responses, and the antibody titer reached high to 1∶8000. Moreover, both two types of immune responses elicited by PCR3.1-hCGβ could attack HeLa cells to induce apoptosis in vitro. The results indicated that the immune responses induced by PCR3.1-hCGβ DNA vaccine have antitumor effects in vitro, and would be helpful to detect the antitumor effects of PCR3.1-hCGβ DNA vaccine in vivo.]]> CHEN Yun,SHI Shu-Qun,YANG Ying and PENG Jing-Pian 1516true ZHENG Bin,WEN Jin-Kun,HAN Mei and ZHOU Ai-Ru 1515true 2O2 for 30 min every 2 days. After total mRNA extraction from the two kinds of cells, the young cells and H2O2-induced premature senesence cells，the cDNA of two kinds of cells were labled by Cy3 and Cy5 respectively were hybridized with microarray containing 4 096 human genes. The gene expression changes were identified by GnePix 4000B and GenePix 3.0. 123 genes changed their expression significantly, which involved in progress of cell cycle,metabolism and protein processing, formation and modification of cytoskeleton and extracellular matrix and signal transduction, and the most interesting finding was that secretory function seemed to be enhanced in premature senescence of fibroblasts.]]> MA Hong,ZHANG Zong-Yu and TONG Tan-Jun 1514true cis-element κB as enhancer, SV40 as basic promoter and neor gene as selective gene. The time and dose effects of d2EGFP and EGFP induced by p65 protein showed that p4κB-d2EGFP was the better NF-κB-responsive GFP reporter gene system because it is more sensitive to detect the changes of gene transcription regulation after p65 vector transiently contransfected with the two vectors respectively. The NF-κB-responsive d2EGFP clonal cell line named HEK-d2EGFP was established after p4κB-d2EGFP stablely transfected into HEK 293 cells. With the cotransfection of NF-κB transcription factor decoy (TFD) and p65 vector into HEK-d2EGFP cells, the results showed the groups of 1 mg/L and 2 mg/L TFD could antagonized the d2EGFP expression induced by p65 protein significantly. It was demonstrated that the NF-κB-responsive d2EGFP reporter system could report and detect NF-κB activation accurately and dynamically.]]> WANG Fu-Long,LIANG Hua-Ping,LIU Xin,XU Xiang and WANG Zheng-Guo 1513true in vivo and it is believed that such interaction is closely related to the sytⅠ physiological function as a Ca2+ sensor in the Ca2+-regulated neurotransmitter release, but the mechanism of the interaction is not clearly understood. Monolayers at an air/water interface combined CD and fluorescence experiments were used to study the characteristic of interaction between C2AB and a phospholipid membrane. The results in the monolayer experiment showed that C2AB domain preferred insertion into the negatively charged phosphatidylserine monolayer and Ca2+ ions were required for the interaction. Electrostatic force was mostly responsible for the insertion of C2AB into PS monolayers. Further CD and fluorescence experiments showed that the secondary structure of C2AB domain in the presence or absence of PS/PC liposome had some relatively small change. The experiments provide useful information concerning the important role of sytⅠ as a Ca2+ sensor in the fusion of secretary vesicles to the plasma membrane, and better understanding the mechanisms of membrane fusion in exocytosis.]]> HE Yu-Hong and SUI Sen-Fang 1512true phoA gene fragment distant from the Asp101-Ser102-Ala103 encoding region to increase the catalytic activity of EAP with a single mutant D101S as parent was directed. Through two cycles of error prone PCR, coupled with a sensitive screening method, an evolved variant 4-186 was obtained. Its catalytic activity was 3-fold higher than that of D101S parent and 35-fold more active than wild-type EAP. The kinetic analysis indicated that the evolved enzyme exhibits a higher substrate binding ability and a higher catalytic efficiency than the D101S parent enzyme. DNA sequence revealed that 4-186 contains two amino acid substitutions, K167R and S374C, both of which locate neither the substrate-binding sites nor the metal-binding sites of EAP.]]> XU Hui-Fang,ZHANG Xian-En,ZHANG Zhi-Ping,ZHANG Yong-Mei and A.E.G.CASS 1511true LI Rong,CHENG Yi-Meng,CHEN Lei,HU Jing,SUN Qiang,HUANG Hong-Yan,WANG Jian and HAN Hua 1510true 33P-dATP and hybridized with the cDNA microarray membrane containing 16 150 genes and ESTs. The hybridized result was confirmed by Northern blot analysis. FLA-3000A Plate scanner and Array Gauge software were used to screen and analyze the expressions of each gene. The background was eliminated and the differences of signal intensity of matched spots over 2 times were set as a marker to identify the differential expression genes. The results suggested that 179 genes were differential expressed, in which 91 genes were up-regulated and 88 genes were down-regulated in NAG7 re-expressed cells. These genes were involved in gene transcription, regulation, proliferation, metabolism, apoptosis, and so on. Northern blot result testified that growth arrest specific protein 1 (gas 1) expressed up-regulated. Especially, the previous results of proteomic research also found that the protein of gas 1 expressed up-regulated. Therefore, the data suggested that gas 1 gene plays a critical role in NAG7 re-expressed HNE1 cells, and provides an important clue to elucidate the mechanism of NAG7 gene.]]> TAN Chen,LI Jiang,PEN Cong,ZHANG Qi-Hong,TANG Ke,LI Xiao-Ling and LI Gui-Yuan 1509true 28～123 plasmids was constructed by gene recombination. The APP28～123 protein was expressed in Escherichia coli and then purified. The purified products were examined for GM1 binding abilities by Western blotting. The effects of GM1 on conformation of APP N-terminus were detected by fluorescence and circular dichroism(CD) techniques. APP28～123 protein could bind with GM1.The intrinsic fluorescence intensity of APP28～123 protein in PC/GM1 vesicles or GM1 solution remarkably increased and the fluorescence peak value blue shifted 20 nm. CD results showed that the major secondary structure of APP28～123 in PBS buffer was α-helix. When APP28～123 incubated with PC/GM1 vesicles or GM1 solution，the α-helix content increased markedly. These results suggested that GM1 might affect the physiological function of APP and change APP span-membrane process and interfere APP trafficking and internalization by anchoring this molecule on the membrane， which may provide much more substrate for γ-secretase and enhance Aβ generation on the cells.]]> DING Ji-Xin,SHA Yin-Lin,RUAN Yan,ZHU Zhong-Jun,HUANG Li-Xin,GENG Hui-Min,NIE Song-Qing and ZHANG Dai 1508true E.coli and purifed by affinity chromatography. Then, ATM was immunoprecipitated from HeLa cells exposed to 10 Gy γ-ray; and the purified GST-p53 was incubated with immunoprecipitated ATM and ［γ-32P］ATP. It turned out that the immunoprecipitated complex of ATM could phosphorylate GST-p53 in vitro. Moreover, the standardization of this method facilitated delineating protein phosphorylation by protein kinase and screening substitutes of other protein kinases.]]> LIU Bin,SONG Yi,DONG Yan and SUN Zhi-Xian 1507true LIU Xin-Ping,DENG Yan-Chun,HAN Jiong,LI Jian,WANG Ji-Cun,LI Ying and YAO Li-Bo 1506true ZHAO Yan,LUO Fei-Jun,TANG Fa-Qing,TANG Min and CAO Ya 1505true Eriocheir sinensis) were examined by whole-cell patch clamp. Under current clamp mode, the depolarization and hyperpolarization were evoked from the three types of neurosecretory cells in response to the GABA (0.1 mmol/L) depending on the Nernst Cl－ potential. Under voltage clamp mode, the inward Cl－ channel currents(IGABA) were resolved from all three types of neurosecretory cells in response to GABA (0.01～5 mmol/L). The GABA currents were activated within 1 200 ms and peaked within 800 ms. No obviously desensitization was observed during GABA application. The dose-response curve showed usual S-shape, with a just-discernible effect at 0.01 mmol/L and near-saturation at 0.5 mmol/L. The GABA currents had reversal potentials that followed Nernst Cl－ potentials when ［Cl－］ was varied. The pharmacological results revealed that the GABA receptor of the crab neurosecretory cells was sensitive to the Cl－ channel blockers picrotoxin and niflumic acid (0.5 mmol/L), insensitive to GABAA receptor antagonist bicuculline and GABAC receptor agonist cis-4-aminocrotonic acid (CACA 1 mmol/L) and trans-4-aminocrotonic (TACA 1 mmol/L).]]> SUN Jin-Sheng,GAO Chun-Lei and XIANG Jian-Hai 1504true Toxoplasma gondii acute infection in serum samples by complete recombinant P35 surface antigen protein, JM109 cell line which can express P35-GST protein was constructed. Then the recombinant protein was separated and purified using affinity chromatography. SDS-PAGE and Western blot were used to analyse the characters of this recombinant protein. Later P35-GST protein was used as antigen to detect Toxoplasma gondii infection by IgM-ELISA. The result of SDS-PAGE showed that the recombinant protein was about 60 ku and was an hydrophile protein. It reacted specifically with Toxoplasma gondii positive serum in Western blot analysis. 60 different serum samples were detected in IgM-ELISA tests using P35-GST as antigen. It was showed that P35-GST can separate acute infection, chronic infection with IgM and IgG positive, chronic infection with IgM negative and IgG postive significantly. P35-GST was very useful in detecting acute and chronic infection of Toxoplasma gondii. It can be concluded that P35-GST can effectively separate acute and chronic infection using serum samples.]]> Lü Bin,WU Shao-Ting,ZHOU Yi-Kai,XU Shun-Qing,ZHANG Ren-Li,GAO Shi-Tong,LIN Min and ZHANG Zhi-Ren 1503true Kip1. Furthermore, over-expression of PKB/Akt suppressed the apoptosis of cells induced by the detachment of the cells from extracelluar matrix. These results suggest the ability of PKB/Akt to promote proliferation and suppress apoptosis in cancer cells.]]> CHEN She,HUANG Chuan-Xin,YIN Xiang-Lei,GU Jian-Xin and SHEN Zong-Hou 1502true WANG Cun-Xin,SONG Wei and CHEN Wei-Zu 1501true DP,PM,EPP,and PIC were calculated for evaluation of forensic application.]]> PEI Li,WANG Jian,JI An-Quan,RONG Liao-Jiang,WANG Hai-Sheng and DENG Hua 1500true lox P site-specific recombinase system, which has two components: Cre recombinase and two 34-bp lox P sites that Cre recognizes, has been widely used in conditional gene knockout/activation to study the structure and functions of gene. In the present study, a cell-permeable fusion protein (His6-NLS-Cre-MTS) containing a 12-amino acid membrane translocation sequence (MTS), a nuclear localization signal (NLS) and an N-terminal His6 affinity tag, was expressed in BL21 strains (e.g., DE3) transformed with pDJHisCre by induction of IPTG. The fusion protein was purified with His-Bond Ni-NTA resin. Its functionality was confirmed in a cell-free recombination assay with a plasmid (e.g., pApoE-SCS-EGFP) containing lox P-flanked gene(s), and in an intracellular recombination system using lox P-flanked STOP cassette-modified BEL-7402 cells, by assaying the expression of enhanced green fluorescent protein (EGFP). This cell-permeable Cre recombinase provides a rapid alternative means of manipulating mammalian gene structure and function in vitro and in vivo. Its advantages and potential uses are discussed.]]> GUO Zhong-Min,XU Kang,YUE Ying,HUANG Bing,TANG Huan,Ma Yun,HONG Xun,CHEN Xi-Gu and XIAO Dong 1499true in vitro and in vivo, P56 inhibites the GR-mediated transcriptional activity.]]> LI Shu-Rong,SU Yong-Ping,LIU Xiao-Hong,LOU Shu-Fen and CHENG Tian-Min 1498true Pleurotus ostreatus by electroporation. Integration of the hITF cDNA into the genome Pleurotus ostreatus was confirmed by PCR analysis. Then competitive enzyme linked immunosorbent assay (ELISA) was developed for detection and quantitative analysis of human intestinal trefoil factor. Results show that the highest hITF protein content in mycelia is 2 000～2 250 ng/g, about 1.5% of the total soluble protein. The transgenic Pleurotus ostreatus is proved to have good biological activity of preventing rats from the gastric ulcer induced by ethanol, in which both of polysaccharides and hITF have a direct effect on ulcer tissue.]]> XU Xiao-Jing,ZHANG Jing,YUAN Yan,YU Mei-Min and RU Bing-Gen 1497true TM PAMAM dendrimers are novel polymers with a molecular architecture characterized by regular, dentritic branching with radial symmetry. Having high density of positive charges on their surfaces in physiological condition because of the protonization of amino groups on the surfaces, and complexing with genetic materials on the basis of electrostatic interactions, those StarburstTM PAMAM dendrimers deliver genes into alive cells.In order to characterize the potential effects of StarburstTM PAMAM dendrimers as a carrier for DNA transfection, six different types generations of StarburstTM PAMAM dendrimers were investigated for their capabilities in binding DNA, and the effects on both DNA transfection and maintenance of cell viability was evaluated in vitro. The experiments demonstrated that it was the full generations but not the half generations of StarburstTM PAMAM dendrimer could transfect eukaryotic cells efficiently. The dendrimer/DNA complexes were very steady, no dissociation of the complexes was detectable in a large scope of pH(2～10). The complexation of StarburstTM PAMAM dendrimer and DNA prevent the reaction that endonuclease dissociates the DNA. In a certain range of dendrimers to DNA charge ratios, the StarburstTM PAMAM dendrimer with higher generations showed much better transfection efficiency than those with lower generations. The transfection efficiency was also variable in different cell lines. StarburstTM PAMAM dendrimers complexing with DNA have no or very low cytotoxicity at the concentrations effective for DNA transfection (≤1.3×10-1 g/L). However, the cytotoxicity of StarburstTM PAMAM dendrimers without binding DNA could be detected at a lower concentration. The results demonstrated that StarburstTM PAMAM dendrimers,as a novel type of low toxicity, non-viral DNA delivery vehicle,had promising potential to mediate DNA transfection in vitro. It provide primary experimental basis for the application of the nanometer material-StarburstTM PAMAM dendrimers in vivo as DNA delivery carrier.]]> GUO Chen-Ying,WANG Heng,LIN Ya-Hui and CAI Qi-Liang 1496true cTnI was linked with TnC by a short DNA sequence coding for 19 neutral amino acid residues. An expression construct for cTnI-C was engineered by inserting the corresponding cDNA into a pET15b plasmid. Then recombinant plasmid was transformed into E.coli BL21(DE3)pLysS cells, and protein expression was induced by isopropyl-β-D-thiogalactopyranoside（IPTG）. Soluable expression of cTnI-C in prokaryotic system was successfully obtained. Fusion protein had an N-terminal His-tag sequence which could be purified by affinity chromatography on a Ni2+-Sepharose column. After one step affinity chromatography the fusion protein shows homogeneity as judged by SDS-PAGE. The fusion protein was stable and easy to be purified,could be used as candidate reference material for cTnI detection systems.]]> YAN Cun-Ling,XU Guo-Bin and XIA Tie-An 1495true Legionella pneumophila and ctxB gene of Vibrio cholerae were PCR amplified respectively. The amplified fusion DNA was ligated to pcDNA3.1(+) vector. The recombinant plasmid, which was named pcDNA3.1-mip/ctxB, was identified by restriction analysis, PCR and further confirmed by sequence analysis. NIH3T3 cell was transfected by recombinant plasmid pcDNA3.1-mip or pcDNA3.1-mip/ctxB with Lipofection strategy. Transient and stable products of mip/ctxB fusion gene was detected by immunofluorescence and Western blot. The results showed that NIH3T3 cell was transfected successfully and stable products can be detected in the transfected cell. To evaluate immunonicity of pcDNA3.1-mip and pcDNA3.1-mip/ctxB, BALB/c female mice were immunized intramuscularly with them and antigen specific antibodies, lymphocyte proliferative response, IFN-γ production and cytotoxic T-lymphocyte response of immunized mice were detected. The results showed that immunogenicity of pcDNA3.1-mip or pcDNA3.1-mip/ctxB immunized mice were higher than control and immunogenicity of pcDNA3.1-mip/ctxB immunized mice were higher than pcDNA3.1-mip immuneized mice. Statistic analysis by one way ANOVA showed that there was significantly difference between groups (P＜0.01).The results provide experimental proof to study of mip/ctxB fusion gene DNA vaccine.]]> WANG Tao,CHEN Jian-Ping,ZHI Ke-Qian,TAO Da-Chang,YANG Chun-Lei and ZHANG Lei 1494true P＜0.05), and that of the right-pawed mice was the highest, but that was decreased in left-pawed mice; after chronic predator stress, in right and ambidextrous mice，the levels of corticosterone were higher than those of the corresponding normal control (P＜0.05)，and the left-pawed and ambidextrous-pawed mices were also higher than those of the corresponding acute predator stress group (P＜0.05). (2) Level of NO. In the acute predator stress group, the level of NO in the supernatants of cultured peritoneal macrophage in right pawed mice was significantly higher than that of left-pawed mice (P＜0.05); after chronic predator stress, those in left, right, and ambidextrous mice were higher than those of the corresponding normal control group (P＜0.05)，and in right-pawed and ambidextrous-pawed mice were also higher than those of the corresponding acute predator stress group (P＜0.05). (3) Level of IL-1β. In the acute predator stress group, the levels of NO in the supernatants of cultured peritoneal macrophage in left-pawed mice were significantly lower than that of the corresponding normal control (P＜0.05); after chronic predator stress, that in right-pawed mice was significantly higher than that of the corresponding normal control group (P＜0.05).In conclusion, brain asymmetry may affect the activity of HPA axis and the functions of macrophages in Balb/c mice exposed to their predator, cat, stress.]]> ZHAO Li-Chun,SU Yun and LI Kang-Sheng 1493true YAN Ge,PU Dan,TANG Ni,GAO Xiao-Ling,SONG Wen-Xin,LU Nian-Fang,WU Gang,TONG-CHUAN HE and HUANG Ai-Long 1492true ZHENG Jing-Min,LIU Zhi-Hong,ZHANG Xin,CHEN Shan and LI Lei-Shi 1491true hPC-1), the complete cDNA sequence from mouse kidney was cloned. The gene was named mPC-1 (GenBank ACC No.AY048852), and its cDNA was 2 193 base pairs. The whole cDNA locus was mapped to mouse chromosome 3A1～A2 by BLAST analysis to the mouse genome. The longest ORF of mPC-1 encoded the putative protein of 224 amino acids, which is 82% identical to hPC-1 coding region, with a coiled-coil domain and PEST domain in each. Bioinformatics analyses show that mPC-1 is comprised of six exons and highly homologous to mD 52 and its first exon is a highly special gene sequence thereof. Special promoter of mPC-1 has been verified experimentally, therefore, it was assumed that mPC-1 is a gene overlapping with mouse mD 52. RT-PCR performed in different mouse tissues or organs and various embryogenesis showed that the expression of the gene was strongly in prostate, kidney and eye, slightly low in stomach and smooth muscle, but not or very weakly in other tissues, however, the mD 52 gene almost expressed ubiquitously in mouse tissues or organs. Consequently, the gene sequence between mPC-1 and mD 52 is markedly overlapping in large-scale but modulated independently. Taken together, the results suggest that mPC-1 is a novel gene in coincidence to hPC-1 either structure or function.]]> LIANG Rui-Xia,ZHOU Jian-Guang,LI Jie-Zhi and HUANG Cui-Fen 1490true in vitro binding assay and co-immunoprecipitation and p93 was showed to co-localize in part with PRX 3 in HEK-293FT cells by immuofluorescence. Taken together, these observations suggest that p93 may participate in various pathophysiologic processes mediated by PRX3 in cardiomyocyte, such as cell growth, differentiation, development，apoptosis and oxygen stress.]]> FENG Yan,LIU Dong-Qing,WANG Zhen,CAO Hui-Qing,SHI Na,DENG Zhong-Duan,DING Jin-Feng and MENG Xian-Min 1489true GUAN Xiao-Xiang,Chen Qi and Fan Le-Ming 1488true PM) between individuals were 1.17×10-36 using allele frequencies obtained from public data on internet and 1.90×10-23 by defining the allele frequencies at each locus as the reciprocal of the number of alleles at that locus. In addition, their genotypes differed from those of the randomly selected LX and HS at 23 and 20 marker loci respectively.]]> HUANG Yin-Hua,XU Gong-Jin,DU Zhi-Qiang,HU Xiao-Xiang,ZHANG Lei,TU Jian-Feng,CHENG Xue-Bo and LI Ning 1487true TU Kang,YU Hui,GUO Zheng and LI Xia 1486true 4 was used to screen phage-displayed peptide library XCX15 and identified a peptide mimetic of Gal-α1,3-Gal. A phage bearing the peptide SCTALSFPSFAFLARGT has been identified to bind human natural antibody strongly. This binding reaction can be competitively inhibited by melibiose. The peptide can also inhibit the human natural antibody-mediated agglutination of pig RBCs.]]> ZHANG Xu,ZHAN Jin-Biao,XU Lin-Hai,YAN Shi-Kun and WANG Ke-Yi 1485true Helicobacter pylori is one of the most common infectious pathogenic agents in human being which causes gastritis, peptic ulcer disease, gastric adenocarcinoma and malignant tumour of the stomach. ure B and hsp A gene were cloned and fused into a shuttle vector pADTrack-CMV. The resultant plasmid was linarized by digesting with restriction endonuclease PmeⅠ, and subsequently were contransformed into E.coli. BJ5183 with an adenoviral backbone plasmid PAdEasy-1. The linearized recombinant plasmid was transfected into adenovirus packaging cell lines HEK-293. The recombinant adenoviruses were typically generated within 7 to 12 days. The recombinant adenoviruses titer was monitored by GFP expression. ELISA was used to test whether adenoviruses efficiently express the Ure B-Hsp A antigen protein in HEK-293. To determine the immunogenicity of the recombinant adenovirus, the delayed type serum IgG, secreted IgA, in the immunized mice were tested. The results showed that both humoral and mucusal immune response in mice were induced.]]> LIU Xiu-Li,LIU Chun-Jie,TAO Hao-Xia,LI Shu-Qin,LI Ji and ZHANG Zhao-Shan 1484true Pichia pastoris -derived recombinant human NTN was injected into the cerebral ventricles 48 h prior to injection of MPTP. Rhesus monkeys in the PD model group acquired PD symptoms that progressed over time, while monkeys treated with NTN had less apparent or no symptoms. Using fluorospectrophotometry, the dopamine (DA), 5-hydroxytry-ptamine (5-HT) and the 5-hydroxyindoleacetic acid (5-HIAA) in substantia nigra, putamen and caudate nucleus in monkeys from the model group were found to be significantly lower than in the normal control group. No significant differences were found between the NTN treatment and normal control groups, but the contents of DA, 5-HT and 5-HIAA in the NTN treatment group were higher than those observed in the PD model group. A dramatic loss of neurons in the substantia nigra in monkeys in the PD model group was observed by light microscopy, while no obvious loss was observed in the NTN treatment group in which the numbers of neurons were similar to those in normal controls. These results indicate that Pichia pastoris -derived recombinant human NTN can prevent PD symptoms as well as protect dopaminergic neurons and preserve DA content in midbrain substantia nigra in rhesus monkeys exposed to MPTP.]]> LI Hong-Jun,SU Ting,MA Yan-Bing,HE Zhang-Long,LU Shuai-Yao,DAI Chang-Bai and SUN Mao-Sheng 1483true P＜0.05) and it was also higher in the left cortex of right-pawed mice than that in the left cortex of left-pawed mice(P＜0.05),（2）The SOCS-3 expression level in right hippocampus was higher than that in the left hippocampus of left-pawed mice (P＜0.05) and it was also higher in the left hippocampus of right-pawed mice than that in the left hippocampus of left-pawed mice(P＜0.05),（3）The SOCS-3 expression level in hypothalamus of right-pawed mice was higher than that of the left-pawed mice (P＜0.05). These results indicate that the SOCS-3 expression level in Balb/c mice brain is associated with brain lateralization.]]> ZHAO Li-Chun,SU Yun and LI Kang-Sheng 1482true LUO Chun-Qing,DENG Li-Bin,ZHOU Jun,LIN Wei and ZENG Chang-Qing 1481true in vitro. The experiments showed that the LPS-binding site of Factor C resides in the S123 region. Even though Cys-rich, EGF-like and Lectin-like domains do not harbor LPS-binding site, the presence of all three domains can improve LPS-binding activity of S123. Recombinant peptide rCES123L containing Cys-rich, EGF-like, S123, and Lectin-like domains exhibits almost the same LPS-binding activity as that of the full-length parental FC. Provided the fact that rCES123L has 4-fold higher production yield than that of recombinant FC in Tn cells, this peptide has a potential in biotechnology and pharmaceutical application for LPS-neutralization.]]> SUN Xiang-Jun,WANG Dong-Ning,ZHANG Wei-Jie and WU Xiang-Fu 1480true 3Vhb, containing cry3A gene and both cry3A and vhb genes respectively, were constructed. Potato leaf-explants were transformed via Agrobacterium harboring pBCry3A or pBC3Vhb respectively. Results from PCR and genomic DNA Southern blotting analysis indicate that cry3A or cry3A+vhb gene have been integrated into the genome of the transformed potato lines with 1～3 copies at least for cry3A gene. The transgenes were stable and expressed even after three successive clonic propagations. Results from ELISA analysis showed that the highest expression level of Cry3A protein reached 0.1% of total leaf soluble protein among cry3A transgenic plants, while that of the cry3A+vhb transgenic plant reached 0.065%. RT-PCR analysis of vhb mRNA and waterlogging test demonstrated that the transgenic potato plants transformed with cry3A+vhb genes expressed vhb gene at least at transcription level and showed obvious higher tolerance against low-oxygen stress. These results together with that of the ELISA for Cry3A indicate that this kind of transgenic plants could be valuble in application for insect resistance and waterlogging tolerance.]]> ZHOU Zhuang-Zhi,ZHOU Yong-Gang,HE Chao-Zu,MANG Ke-Qiang and TIAN Ying-Chuan 1479true XKS1 was cloned from Saccharomyces cerevisiae NAN-27 and ligated into plasmids pMA91 and YEp24, producing pMA-xy203 and YEpP-xy204, respectively. In both plasmids, XKS1 was under the control of PGK promoter. pMA-xy203 was transferred into the pre-constructed recombinant yeast strain H158-XR-XDH, which contains the XYL1 and XYL2 genes from Pichia stipitis, encoding xylose reductase and xylitol dehydrogenase respectively, in an episomal plasmid vector. This new recombinant strain was named HSXY-251. YEpP-xy204 was transferred into the pre-constructed recombinant yeast strain H158-XI, which contains the xylA gene from Thermus thermophilus encoding xylose isomerase in an episomal plasmid vector, resulting in recombinant strain HSXY-252. The xylulokinase activities in HSXY-251 and HSXY-252 were respecfively 14 and 6.7 times higher than that in the parent strain. Glucose and xylose co-fermentation carried out with HSXY-251 under oxygen-limited conditions at 30℃ resulted in 9.4 g/L ethanol concentration with 12.4 g/L xylose consumed. Xylose consumption and ethanol production were respectively 120.9% and 36% higher than in the parent strain. Furthermore, the by-product xylitol was 0.7 g/L, a decrease of 84.9%.]]> SHEN Yu,ZHENG Hua-Jun,WANG Ying,BAO Xiao-Ming,QU Yin-Bo and BAI Feng-Wu 1478true CHAI Li-Hong,CHEN Ji-Shuang and CHEN Zi-Yuan 1477true DU Zhan-Wen,ZHANG Xin and ZHANG Jun-Wu 1476true neo gene were obtained. The β-galactosidase activity was detectable in 14 ES colonies. ES cells from 3 trapped lines were introduced into blastocysts by microinjection. Two chimeric mice lines were generated. One trapped mutation went through the germline. Genomic sequences adjacent to integration sites of the constructs were isolated by plasmid rescue. The results of sequencing suggested that the trapped gene is possibly a novel gene. The expression pattern of this gene shown by β-galactosidase expression was restricted in abdomen and lib bud of E10.5 mouse embryo.]]> TAN Xiao-Hong,CHENG Xuan,MAO Chun-Ming,CHEN Guang-Hui and YANG Xiao 1475true A392/A280) was developed to survey the purity of the P450cam mutant protein during the purification of the mutants protein. Thus the course of the purification became compact and the efficiency of the purification had been improved. β-Mercaptoethenol had been developed to maintain the deoxidization of the protein during the purification. Crystals of the F87L/Y96F/L244A/V247A mutant were grown by vapor diffusion method. X-ray diffraction data were collected to 0.22 nm resolution on a Mar Research area detector in house. The structure was determined by Difference Fourier Method. The final crystallographic R factor and Rfree factor are 0.197 and 0.247 respectively. The RMS deviations of bond length and angle of the mutants are 0.001 77 nm and 1.96°respectively. Structure comparison indicates that there is no obvious conformaional change between P450cam and the F87L/Y96F/L244A/V247A mutant. After mutation, the structure of the active pocket became larger but its hydrophobility increased, which are consistent with the aim of mutantion]]> LIU Bin-Bin,SUN Lei,XU Feng,LOU Zhi-Yong,PANG Hai,L.L.WONG and LIU Yi-Wei 1474true in vivo study of its normal function and the role of BPOZ in tumorigenesis. The mouse genomic DNA sequence of BPOZ gene was verified through bioinformatic analysis. According to the BPOZ genomic DNA sequence, the strategy of gene targeting and constructing of knockout vector (XpPNT-BPOZ) were established to delete the region of mouse genome, which spans from exon 2 to 12 of BPOZ gene. The gene knockout vector XpPNT-BPOZ was constructed and confirmed by restriction enzyme digestion and sequencing. Electroporation of ES cells with XpPNT-BPOZ and screening of both G418 and Ganciclovoir resistant clones were performed according to common protocol. The homologous recombined ES cell clones were identified by PCR and confirmed by Southern blot analysis. After transplantation of homologous recombined ES cells into blastocysts through microinjection and mating of chimeras with C57BL/6J mice, 30 offspring with Aguoti fur in color were acquired. 15 of them (50%) show genotype heterozygous for BPOZ. The BPOZ heterozygotes were intercrossed to generate mutants homozygous. Finally, the cohort of mutants homozygous for BPOZ was established. The mutants are fertile, and further phenotype analysis is under way.]]> XIANG You-Gui,SUN Xia,WANG Long,YAN Lan-Zhen,YANG Hua,LIU Wei,XU Yong,XU Guo-Jiang,WANG Yi,FEI Jian,FU Ji-Liang and WANG Zhu-Gang 1473true Bombyx mori) grew gradually until 2 days before emergence, and markedly enlarged due to the accumulation of glue like substances (mainly including 85% water and 11% proteins). However, the Ng mutant female moth only secreted a small amount of glue-like substance and laid loose eggs naturally. High-resolution two-dimensional polyacrylamide gel electrophoresis, followed by computer-assisted analysis was used to screen the proteins pattern between normal and Ng mutant colleterial gland to find quantitative and qualitative difference in proteins expression. Protein spots were resolved in the secretory region of colleterial gland of silkworm and more than 700 protein spots were resolved and most of the proteins were distributed in the area from 30 ku to 70 ku and pH 4～8. Through the comparison and analysis, it was found that 3 proteins were only expressed in the later pupae stage and moth stage. However, these proeins have no expression in the Ng mutant especially spot No.2 and 3. These differentially expressed proteins were actin identified by matrix-assisted laser desorption/ionization-time of flight mass spectrometry. The results indicated that the actins participate or regulate the exocytosis of colleterial gland and other differential expressed proteins might be having some relations with the glue-like proteins secretion.]]> JIN Yuan-Xiang,XU Meng-Kui,CHEN Yu-Yin and JIANG Yong-Huang 1472true HU Wei,XIAO Zhi-Qiang,CHEN Zhu-Chu,LI Jian-Ling,ZHANG Peng-Fei,FENG Xue-Ping,YI Hong,YU Yan-Hui,TANG Xin-Ke,LIU Qing-Ping and LIANG Song-Ping 1471true Magnaporthe grisea. Of 37 cDNA clones initially identified by reverse Northern blot, 5 clones were chosen as probes for further Northern test. The result showed that all of these 5 clones were expressed both in highly, moderately resistant and susceptible rice lines in a pathogen inducible manner. According to the homology analysis, these clones were suggested to be involved in plant defense response by: (1) inhibiting growth of fungus and even causing its death; (2) excluding toxic compounds delivered by fungus; (3) priming signal transduction; (4) and/or regulating metabolic activities in agreement with plant defense.]]> LUO Wen-Yong,LI Xiao-Fang,HU Jun,LIU Wen-Hua,XIAO Xin and LIU Liang-Shi 1470true LI Chun-Bo,GAO Feng,ZHONG Yong-Wang,WEI Chun-Hong and LI Yi 1469true Cinnamomum camphora). They are named cinnamomin and cinphorin. The molecular mass of cinphorin A-chain is only half of cinnamomin A-chain, while their B-chains are the same. Their A-chains are RNA N-glycosidases, and the B-chains are lectins. The intrinsic cytotoxicity of type Ⅱ RIP is greatly dependent on the carbohydrate-binding activity and the specificity of its B-chain. The lectin activity of cinnamomin and cinphorin are investigated and compared with each other. They showed similar hemagglutination activity. Their saccharide binding specificities were studied by hapten inhibition, indicating that they were both galactose-specific. However, N-acetylgalactosamine failed to bind to the two RIPs as ricin/abrin did. The interactions of the two RIPs with specific saccharides were also investigated by fluorescence spectroscopy through which association constants were obtained. Their association constants of galactose or lactose were found to be identical.]]> YANG Hai-Yan,LI Rong-Xiu and LIU Wang-Yi 1468true E.coli cells. This His-tagged DHFR could be purified routinely by Ni-NTA agarose resin and the His-tag could be removed by thrombin easily. This engineered DHFR has the same enzyme activity as the enzyme without His-tag obtained by iso-electrophoresis.]]> ZHU Zhi-Yong,TAN Bo,ZHANG Wen-He and ZHANG Hong-Jie 1467true in vitro differentiation of P19 cells into cardiomyocytes was established using RT-PCR. GATA-4 is expressed at day 2 after treatment with DMSO, and Nkx2.5 at day 3. hhlim is almost undetected until day 4 after DMSO treatment. Western blot had the same results, which showed that hhlim might participate in the cardiac differentiation pathway. P19-hhlim-overexpressing cells did not show morphological or biochemical evidence of cardiomyogenesis without DMSO treatment, but overexpression of hhlim had a benefit effect on cell aggregates that were especially obvious in the absence of DMSO. Moreover, when hhlim-overexpressing cells were treated with DMSO, the expression of Nkx2.5 and GATA-4 appeared earlier compared with the control. hhlim-deficient P19 cells induced by DMSO retained the ability to differentiate into cardiomyocytes. The time course of expression of Nkx2.5 and GATA-4 in the hhlim-deficient clones was significantly reduced, whereas these clones treated by RA produced neuroectodermal derivatives. Then it was concluded that lack of hhlim expression specifically inhibits cardiomyocyte formation of P19 cells without affecting neuronal differentiation pathway. These data showed that overexpression of hhlim could enhance cardiogenesis of embryonic stem cells whose mechanism might be involved in upregulation of the expression of Nkx2.5 and GATA-4.]]> ZHENG Bin,WEN Jin-Kun,HAN Mei and ZHOU Ai-Ru 1466true 6 clones. To ascertain the interaction, the 51st clone which interacted more intensive with CEP65 in yea was selected to do GST pull-down assay with CEP65. The result indicated that CEP65 also interacted with the 51st clone in vitro. Bioinformatics analysis showed the interaction between CEP65 and the 51st clone maybe regulate the cell differentiation and proliferation by WW domain or protein kinase C.]]> JIN Geng-Lin,SUN Yu-Ning,ZHANG Jian-Zhi,MA Hong,MENG Lin,WU Jian and SHOU Cheng-Chao 1465true in vitro. First, the BALB/c mouse ES cells were cultured for 5 days to develop into EBs in EBs culture medium. Then, the cell growth factors such as transform growth factor (TGF), basic fibroblast growth factor (bFGF) and hepatocyte growth factor (HGF) were added into the medium to induce the differentiation of EBs cells into hepatocytes. During the culture medium, the reverse transcriptase-polymerase chain reaction (RT-PCR), immunocytochemistry (ICC) and radioimmunoassay (RIA) were used to determine the expression of hepatic genes and proteins, such as α-fetoprotein (AFP)，albumin (ALB)，glucose-6-phosphatase (G6P)，tyrosineaminotransferase (TAT)，cytokeratin 8 (CK8), cytokeratin 18 (CK18) and urea, to analyse the hepatic differentiation. At last, the hepatic differentiation ratio was also determined by counting the ICC-positive cells. In results, the mRNA of AFP, ALB, G6P and TAT didnt respectively expressed till at day3, day9, day11 and day13. AFP, ALB and urea in culture medium were not detected till at day8, day11 and day12 with a concentration of (3.4±0.6) μg/L, (0.21±0.04) mg/L and (8.3±1.4) μmol/L. Hepatic proteins such as AFP, ALB, CK8, CK18 didnt respectively expressed in cytoplasm till at day7, day9, day9, day11 and ICC-positive cells had morphological structures consistent with mouse primary hepatocytes. Hepatic differentiation ratio was determined at day19 with ratio of 32%. In conclusions, hepatocytes could be obtained in ES cells differentiation system and get matured with differentiation ratio of 32% when growth factors such as TGF, bFGF and HGF added into culture medium. This may produce a new resource of hepatocytes in liver engineering and hepatocytes transplantation for treating hepatic failure.]]> HU An-Bin,CAI Ji-Ye,ZHENG Qi-Chang,HONG An,HE Xiao-Qing,DAI Yun,SHAN Yu,PAN Yun-Long and LI Ling-Song 1464true in vitro. Transfected B16 cells by PA317 carrying the pLNCX and untransfected B16 cells were used as control. The B16pLNCX-RI cell line with a stably high expression of RI was identified by PCR, RT-PCR, Western blot and immunofluorescence assay respectively. The results showed that the transfected RI gene might significantly inhibit cell proliferation, migration, and enhance cell adhesion, as well as, make morphological changes in vitro. Cell doubling time were (24.98±0.16)h, (25.62±0.28)h, (32.64±1.11)h in B16 cells, B16 pLNCX and B16 pLNCX-RI cells respectively. Cell adhesion rate was significantly increased by 19.5% and 17.8% as well as cell migration was reduced by 60% and 61.4% in B16 pLNCX-RI cells compared with pLNCX B16 cells and B16 cells respectively. B16 pLNCX-RI cells became flatter, less nucleoli, less division phases and weaker alkalophilic quality of cytoplasm compared with control groups, which should imply that cell proliferation viability was decreased and malignant phenotype was improved on the cell transfected RI. Mice injected with B16 pLNCX-RI cells show a significant inhibition of the metastasis of tumor with lighter lung weight, fewer metastasis nodules, a lower incidence rate, a lower density of blood vessels and longer survival with respect to the control groups, which implied that RI might be involved in metastasis of melanoma. The results of experiments show RI has a significant antitumor metastasis effect and suggest that it is partially responsible for inhibiting angiogenesis, decreasing cell proliferation, reducing cell migration and enhancing cell adhesion.]]> CHEN Jun-Xia,TIAN Yu-Xiang,FU Pan-Feng,XIA Jun,YAN Ping and CUI Xiu-Yun 1463true YU Jing,DENG Yan-Qiu,YANG Ying,ZHANG Jia-Yu,ZHANG Ye-Ping,ZHANG Shao-Hua and WANG Jian-Zhi 1462true PDX-1) so as to study the expression and effect of PDX-1 gene in bone marrow mesenchymal stem cells, human PDX-1 gene was ligated into shuttle vector pAdTrack-CMV. Homologous recombination was performed in BJ5183 bacteria by cotransforming linearized shuttle plasmid with adenovirus backbone plasmid pAdEasy-1.The recombinant plasmid was packaged and amplified in 293 cells.Mesenchymal stem cells(MSCs) were isolated from healthy adult bone marrow.The adenovirus was transfected into MSCs. The recombinant adenovirus vector has been successfully constructed according to the results of sequencing,PCR and enzyme digestion identification.The expression of PDX-1 and insulin in transfected MSCs at 7 days was detected by RT-PCR and immunocytochemistry. The value of insulin secretion was (15.21±3.50 )mIU/L from the transfected cells at 7days.These results showed that PDX-1 gene modified MSCs could be the beta cell replacement for those diabetes patients who need insulin-secreting cells.]]> Li Yan-Hua,Zhang Rui,Wang Yun-Fang,Zhao Lian-Xu,Chen Lin,Yue Wen and Pei Xue-Tao 1461true TANG Chao-Ke,XI Shou-Min,YIN Wei-Dong,HE Xiu-Sheng,LIU De-Ping,YI Guang-Hui,Yang Jun-Hao,Wang Shuang,WEI Xing and YANG Yong-Zong 1460true Selnocosmia huwena. HWTX-Ⅴ contains 35 amino acid residues, including six cystein residues that form three pairs of disulfide bond. The positions of the disulfide bonds of HWTX-Ⅴ have been determined. To map the disulfide bonds, native HWTX-Ⅴ was multi-proteolytically digested. The resulting peptides were identified by matrix assisted laser desorption/ionization time of flight mass spectrometry, and it indicated the presence of one disulfide bond Cys9-Cys21. The partially reduced peptides by using Tris- (2-carboxyethyl)-phosphine at pH 3.0 over 12 minutes were purified by reverse phase high-performance liquid chromatography, and then one -free thiod and two-free thiod fractions were collected. The free thiods were carboxamidomethlate by iodoacetamide at the concentration of 0.5mol/L.The locations of disulfide bond Cys2-Cys16 and Cys15-Cys28 were proved by comparing N-terminal sequencing analysis these partially reduced and alkylated HWTX-Ⅴ with that of the intact peptide. Finally, the three disulfide linkage of HWTX-Ⅴ could be assigned as Cys2-Cys16,Cys9-Cys21 and Cys15-Cys28.]]> ZHANG Peng-Fei,XIAO Shun-Yong and LIANG Song-Ping 1459true E.coli T-protein, protein limited digestion and fragmentation cloning were employed. Fragment 1～93 and fragment 96～373 were cloned and expressed according to the results of limited digestion of T-protein respectively. Two fragments were found to have chorismate mutase and prephenate dehydrogenase activities respectively. In a conclusion, E.coli T-protein do have independent domains, N-terminal 93 amino acids belongs to chorismate mutase and C-terminal 277 amino acids belongs to prephenate dehydrogenase.]]> CHEN Jing and CHEN Shu-Qing 1458true ZHANG Ai-Qun,WU Yan,ZHOU Ning-Xin and YEW David Tai-Wai 1457true LIU Rong,WANG Xiao-Chuan,HUANG Yao,HE Zhi-Song,CHU Shu-Juan,TANG Yun,TIAN Qing and WANG Jian-Zhi 1456true LIN Jue-Long,XU Li-Yan,LI En-Min,CAI Wei-Jia,NIU Yong-Dong,FANG Kun-Yang,XIONG Hua-Qi,SHEN Zhong-Ying and ZENG Yi 1455true PC-1 was identified as a novel gene that expressed higher in the much more malignant human prostate cancer cell lines and its translation product meets some points of transcription factors. In order to study the transcriptional activation activity of PC-1，firstly, the yeast two-hybrid system was used and the whole length as well as various regions of PC-1cDNA were cloned into the expression vector pAS2-1 and transfected into the yeast cell line CG-1945 respectively, the result of the reporter genes lacZ and His3 activation assay showed that PC-1 had transcriptional activation activity and this activity was mapped within its N terminal 46 amino acids. Then，the various regions of PC-1 cDNA were cloned into the expression vector pZHO1 and cotransfected with the pTRE-luc plasmid of the reporter gene into COS7 and C4-2 mammalian cells, the 46 amino acids in the N terminal of PC-1 were found to have transcriptional activation activity by the firefly relative luciferase activity assay.]]> ZHANG Hao,ZHOU Jian-Guang,LI Jie-Zhi,YU Mei and HUANG Cui-Fen 1454true in vitro was set up, and the redistribution of α3,α5,β1 integrin in H727 cells subjected to 24 h cyclic strain was analyzed. Human pulmonary epithelial H727 cells seeded on either fibronectin or collagen Ⅳ surfaces were subjected to 15% elongation at a frequency of 40 cycles/min. Confocal microscopy revealed that α3,α5,β1 integrin in H727 cells were concentrated and brighter,and cells created a large fusion of focal adhesion or adhesion plaques after 24 h exposure to strain. Under static condition, α3,α5,β1 integrin in H727 cells maintained a diffuse pattern with an occasional speckle concentration in some regions. Furthermore α3,α5,β1 integrin in H727 cells transferred from the apical layer to the basal layer and created adhesion plaques, and affinity between integrins and extracellular matrices(ECM) was enhanced. It is concluded that the formation of effective focal contacts is the concerted interaction between receptors recruitment and ligand occupy. This interaction plays an important role in transducing mechanical stimuli into intracellular signals.]]> ZHANG Hui-Jing,YANG Li,CAI Shao-Xi,LU Xiao and WANG Yuan-Liang 1453true TAN Yun-Nian,TAO Yong-Guang,LI Li-Li,LIU Su-Fang,TANG Min,GU Huan-Hua and CAO Ya 1452true in situ from EM data of the ultrastructure of the nucleolus in allium sativum cells and from the modified NAMA-Ur DNA specific staining method was developed. For this purpose, a new methodology of multi-scale morphological gradient operator and automatic analysis software named “HEREN.CELL” were created and adopted. By using this method, superfine contour of the nucleolar ultrastruture was obtained and showed the location and location of nucleolar DNA in situ. From the results, a “petaline envelope” model was proposed, which indicated that nucleolar DNA fibrils radiate from center to periphery of the nucleolus. The study will contribute to the development of the EM image processing technology and the study of nucleolar ultrastructure of the nucleolus.]]> CAI Nian,HU Kuang-Hu,TAO Wei,SU Wan-Fang,LI Fang-Zhen and HU Ying-Xiong 1451true MA Jing-Yun,CAO Yong-Chang and BI Ying-Zuo 1450true Ercc6l (excision repair cross-complementing rodent repair deficiency, complementation group 6-like) by MGNC. The cDNA of Ercc6l was 4 002 bp and firstly cloned in silico and then identified by RT-PCR in mouse embryo heart. This gene was composed of 2 extrons and mapped to mouse chromosome X. The longest ORF encoded a putative protein of 1240 amino acids. Eight highly conserved sequence regions of SNF2 protein were present in the deduced protein. Alignment against members of SNF2 family initially placed Ercc6l into ERCC6 subfamily of SNF2 family. EGFP-tagged Ercc6l protein was localized in cytoplasm of HeLa, NIH3T3 and B16 cells. BLAST search in EST database retrieved 69 homologous ESTs of Ercc6l, most of which were from mouse embryonic and tumorous tissues. RT-PCR performed on a panel of different mouse tissues demonstrated that Ercc6l strongly expressed in mouse embryo but significantly downregulated in neonate and adult normal tissue. These results suggest that Ercc6l is a novel member of SNF2 family and may be crucial for development of embryo and tumor.]]> CHEN Xiang-Gui,LI Yong,ZANG Ming-Xi,PEI Xin-Rong,XU Ya-Jun and GAO Li-Fang 1449true HU Jun and ZHANG Jing 1448true FENG Xiu-Cheng,QIAN Min-Ping,DENG Ming-Hua,MA Xiao-Tu and YAN Xi-Ting 1447true Plutella xylostella larvae by using β-1,3-glucan affinity precipitation method. The proteins were present in plasma but absent in haemocytes. The β-GBPs had a specific affinity to β-1,3-glucan, a prophenoloxidase (ProPO) trigger in P.xylostella hemolymph. The phenoloxidase (PO) activity induced by the coexistence of both β-GBPs and β-1,3-glucan was significantly higher than that by the β-GBPs or β-1,3-glucan alone. Moreover, the PO activities induced by mycelial lysates of four Zoophthora radicans isolates at the presence of β-GBPs were much higher than that triggered by their protoplast lysates. The results indicate that the β-GBPs may trigger the ProPO activity in P.xylostella hemolymph only if they specifically recognized β-1,3-glucan from Z.radicans cell walls. This is a proof for a hypothesis that entomophthoralean protoplasts may evade from insect host immune defense. The protoplast or mycelial lysates of different Z.radicans isolates differed in ability to trigger the PO activities in the following order: ARSEF1342＞ARSEF2699＞F99101＞ARSEF1100. This trend was in accordance with that of their virulence to P.xylostella larvae, suggesting a possible relationship between host immune defense and fungal virulence.]]> LIU Qing-E,XU Jun-Huan and FENG Ming-Guang 1446true In situ hybridization (ISH) and immunohistochemistry (IH) suggested that it express in dentritic cells of NP and NPC tissues. It is meaningful to do further studies on the reason of low expression in NPC tissue and the function of NAP1 gene.]]> LI Feng,JIANG Wei-Hong,YANG Xu-Yu,YIN Zhi-Hua,FENG Xiang-Ling,LIU Wei-Dong,WANG Lei,ZHOU Wen and YAO Kai-Tai 1445true YANG Jie-Lin,ZHU Nai-Shuo,WANG Ying,GUAN Xiao-Feng and ZHENG Zhao-Xin 1444true Bombyx mori) embryo by introduction of double-stranded RNA corresponding to the silkworm Bmwh3 gene into the preblastoderm eggs of wild-type has been established based on the previous research. The results showed that the RNAi was successful when injected Bmwh3 dsRNA into the preblastoderm eggs within 8h after laid of wide-type silkworm and the concentration of the Bmwh3 dsRNA must be high to 2.0 g/L. It can also induce the other w3 mutant phenotype——translucent larvae when injected Bmwh3dsRNA into the third day of developed eggs. A conclusion can be primarily drawn that the Bmwh3 is not only involved in the transport of the precursors of the ommochrome and pteridine pigments to the eye and egg，but also involved in the transport of the precursors of the pigments to the larvae skin.]]> LIU Chun,SHUAI Xiao-Rong,CHENG Ting-Cai,XU Han-Fu,LI Chun-Feng,DAI Fang-Yin,XIA Qing-You and XIANG Zhong-Huai 1443true geo as the reporter, and gives no discrimination on the types of trapped genes. A new reporter gene named peo was obtained by in turn fusing Exotoxin A of Pseudomonas aeruginosa, 2A sequence, transmembrane region of IL-2 receptor and neomycin phosphotransferase genes, Peo is able to report genes encoding secretory proteins specifically. To prove peo's specificity for secretory proteins, three plasmid vectors, representing three types of sequences possibly trapped by gene trap vector during screening, were constructed based on peo reporter. After transfecting HeLa cells with these plasmids, followed by G418 selection and PLAP staining, it is shown that the new vector could distinguish secretory proteins from others, and the gene trap vector taking peo as the reporter can trap secretory protein genes efficiently.]]> SUN Qiang and HAN Hua 1442true +/K+ ATPase beta-3 subunit (ATP1β3), phospholipase C delta 1 and epididymal secretory protein. The significance of identification of these proteins may provide mechanistic insight into the biologic role of HPO in testis.]]> CHEN Xiao-Xiao,LI Yong,QIU Zong-Yin and HE Fu-Chu 1441true in vitro fertilization and parthenogenetic activation, γ-tubulin concentrated around the male and female chromatin and pronuclei. It is also found to be localized in sperm acrosomal cap and the neck. During early cleavage, γ-tubulin distributes around the blastomere nuclei. The results suggest that γ-tubulin is a major regulator of microtubule assembly in porcine eggs and that both oocyte and sperm contribute centrosome materials to fertilized eggs.]]> YAO Li-Juan,ZHANG Li-Sheng,KOU Zhao-Hui,CHEN Da-Yuan and SUN Qing-Yuan 1440true LI Ya-Li,ZOU Wei,MA Ke-Li,XIA Quan and CUI Zhao-Chun 1439true 2 (cPLA2) in this process. Cell viability was determined by MTT assay, cell apoptosis was assessed by phase-contrast microscopy, fluorescence microscopy and flow cytometry (FCM), PLA2 activity was determined by measuring the release of 3H-arachidonic acid (3H-AA) from prelabeled cells, phosphorylation of cPLA2 was analyzed using Western blot, changes of ［Ca2+］i in single cell were monitored by laser scanning confocal microscope (LSCM). The results showed that MM-LDL (100～300 mg/L) induced decrease in cell viability in a dose- and time- dependent manner. After 48 h exposure to 300 mg/L MM-LDL, apoptosis with cell shrinkage, chromatin condensation and nuclear fragmentation were observed. FCM assay showed that the apoptotic ratio rose with an increase in concentration of MM-LDL. MM-LDL caused a rapid elevation of ［Ca2+］i in HUVECs partly through calcium influx. Incubation with MM-LDL induced calcium-dependent cPLA2 activation companied with its phosphorylation. Inhibition of cPLA2 activity with 5 mmol/L EGTA or 15 μmol/L AACOCF3 reduced MM-LDL-induced apoptosis by 41.8% and 33.6%, respectively. Addition of exogenous AA (50 μmol/L) reversed AACOCF3-induced reduction of apoptosis. It was demonstrated that MM-LDL induced apoptosis in HUVECs. cPLA2 activated by increase in intracellular Ca2+ was involved in the signal pathway.]]> WANG Yun,ZHOU Xin,WANG Bing-Hua,CHEN Li-Da,ZHANG Ji and CAO Jin-Xiu 1438true Trametes gallica by using cDNA microarray from Phanerochaete chrysosporium, a well known model strain used for studying lignin-degrading systems. The results showed that there were only 172 positive clones detected on the microarray and 95.9% of the clones had a ratio of Cy-5/Cy-3 from 0.5 to 2.0. However, only 3 and 4 clones with the Cy-5/Cy-3 ratio over 2.0 and lower than 0.5 respectively, were found, which are corresponding to 5- and 12-day cultures of T.gallica. One hundred and twenty-two clones were randomly selected and sequenced, of which 118 clones could be perfectly located in the genome of P.chrysosporium. The result indicates that there is obvious sequence difference between probes from T.gallica and inserts on the chip from P.chrysosporium, exhibiting far relative between these two fungi. Two interesting clones have been obtained from homology comparison, one corresponding to a fragment of peroxidase lpoB gene of P.chrysosporium and the other encoding a sort of heat shock protein.]]> SUN Xun,JIANG Ming-Feng,LI Xiao and ZHANG Yi-Zheng 1437true Mycobacterium tuberculosis (MTB) isoniazid (INH) resistant strains, and 32 protein spots were found to express differentially. Five protein spots which remarkably increased in U937 infected with MTB INH-resistant strains were measured by matrix assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS). The data obtained from peptide mass fingerprinting were used in protein database search. Five protein spots in gel were identified as HSP105β, inhibitor of apoptosis protein-1, phosphoglycerate mutase 1, cathepsin B, desmocollin 3. These data provide insight into the changed global protein patterns of the U937 while in stasis shortly after infection and may prove useful for further study in the interaction between MTB INH-resistant strains and host.]]> LIU Li-Rong,YUE Jun and WANG Hong-Hai 1436true Pseudomonas exotoxin A based immunotoxins against carcinoembryonic antigen (CEA) were constructed and named CEA(Fv)/PE38/KDEL and PE35/CEA(Fv)/KDEL. Antigen binding activity of the immunotoxins were assessed by flow cytometry, internalization of immunotoxins was detected by optical confocal microscopy and flow cytometry quantitatively. Cell apoptosis and necrosis were analyzed by staining the cells with FITC-annexin V/PI. The MTT method was used to assay the cytotoxicity of the immunotoxins. The biological functions of the two forms of immunotoxins in the different functional phases were compared. And the interaction between different functional phases was analyzed. The results revealed the whole functional process from the cell binding to the cytotoxicity of the immunotoxins, which provided more details about the biological functions of immunotoxins and could be valuable in the mechanism research and the optimization of immunotoxins.]]> HE Dan,YANG Hui,CHAO Kai,LIN Qing and HUANG Hua-Liang 1435true Shan Yu,Chen Xi-Gu,Huang Bing,Hu An-Bin,Guo Zhong-Min and Huang Wen-Ge 1434true E. SüLEYMANOLU 1433true in silico by aligning dozens of overlapping rat and chicken ESTs and cDNA sequences identified from GenBank. Human MOB was mapped on chromosome 10q11.1～11.2. Homology searches with the deduced 413 amino acid residues revealed human MOB shares 97% similarity with murine MOB, 97% with rat MOB, 91% with chicken MOB and 45%～73% with lots of hypothetical proteins of various origin. The predicted protein contains SAM domain, which has been suggested to be an evolutionarily conserved protein-binding domain that is involved in the regulation of numerous developmental processes in diverse eukaryotes and can potentially function as a protein interaction module through its ability to homo- and heterooligomerise with other SAM domains. Homology searches and domain query indicate that human MOB is a member of potential phylogenetically conserved MOB family. RT-PCR revealed that human MOB was almost expressed in all kinds of tissues and cells, which is completely consistent with the in silico expression pattern identified by bioinformatics analysis. In contrast to bioinformatics analysis and previous study, however, human MOB has widespread subcelluar expression, primarily in nuclei. Further study in HeLa cells shows that overexpression of human MOB seems not to influence cell cycle and apoptosis.Taken together, human MOB is a novel phylogenetically conserved gene, which is proposed to express in almost all tissues and cells, and has widespread subcelluar expression, primarily in nuclei. It may play a role in signal transduction and thus be involved in development regulation, but seems not to influence cell cycle and apoptosis. Further investigation remains needed.]]> YUAN Hong-Feng,WANG Dong-Mei,LI Hai-Min,CHEN Lin,BAI Ci-Xian,YUE Wen and PEI Xue-Tao 1432true YUAN Can,ZHANG Hua-Li,LIU Ying,WANG Qiu-Peng and XIAO Xian-Zhong 1431true ZHANG Zhi-Zhen,DUAN Lian and LI Kang-Sheng 1430true 0 mice produced through microinjection. The copy number ranges from 1 to 8, and the integration rate is 11.7%. Human α-lactalbumin have been detected in milk samples of the four founder female transgenic mice by SDS-PAGE gel electrophoresis and Western blotting, and the concentrations of the human α-lactalbumin are quantified to be 0.62g/L, 0.48g/L, 0.56g/L, 3.21g/L respectively by radioimmunoassay (RIA) method. Meanwhile, one offspring of the No.50 male transgenic mice has also expressed the human α-lactalbumin, and the concentration is 1.03 g/L in the milk. The transgenic construct herewith has the characterization of a high-level expression and stable inheritance. A foundation has been provided for improving the quality of cow milk thorough transgenesis of human α-lactalbumin.]]> YU Shu-Yang,CAO Geng-Sheng,FAN Bao-Liang,LI Ning,DUAN Ying-Li,DAI Yun-Ping and WANG Li-Li 1429true CHEN Su-Hong,ZHANG Min-Li,HUANG Jian,DING Yu,BO Xiao-Chen and WANG Sheng-Qi 1428true Escherichia coli in combination with Western blotting, two linear epitopes were identified on non-structural protein 3ABC of foot-and-mouth disease virus(FMDV), which covering amino acid residues 106～155 and 156～190 on 3ABC. The epitopes gave positive reaction with sera from pigs or guinea pigs infected with different serotypes of FMDV, but not with sera from vaccinated or naive animals. The method based on expressed peptide for mapping epitope on viral protein was reliable and applicable.]]> SUN Tao,LU Ping and WANG Xin 1427true ZHANG Hao,ZHOU Jian-Guang,LI Jie-Zhi and HUANG Cui-Fen 1426true E.coli BL21(DE3). The recombinant protein was expressed efficiently in E.coli BL21(DE3,pET-N58) as inclusion body with a yield of about 35.4% of the bacterial total protein. The partial purified recombinant protein inhibits the proliferation of human umbelical vein endothelial cells and the CAM angiogenesis. The recombinant protein also suppresses the growth of primary B16 murine melanoma in C57BL/6 mice.]]> CHEN Hong,ZHANG Tian-Yuan,LUO Jin-Xian and HU Zhi-Shang 1425true EF-1α) for transcription of genes of interests, and mouse dihydrofolate reductase (dhfr) gene under control of SV40 promoter (PSV40) for clonal selection and amplification was first constructed . The vector was named pED5. The expression efficiency difference between pED5 and pCdhfr1, a vector utilizing CMV enhancer/promoter (PCMV-IE) for foreign protein production, was analyzed using human interferon-β (IFN-β) gene and human secreted alkaline phosphatase (SEAP) gene as reporters. When analyzed in transient expression, pED5 showed a little more protein produciton than pCdhfr1. However, in continuous expression, when serum concentration was lessened to slow down cell proliferation, pED5 expressed 3.1 times more reporter proteins than pCdhfr1, which implied that PEF-1α was less affected by cell cycle status in contrast to PCMV-IE, making pED5 a good expression vector for foreign protein production.To further enhance protein productivity of expression vectors, two artificial transcription activating domains, AH and VP2, were linked to cI repressor protein of phage λ through a soft linker, respectively, and thus two artificial transcription activators were created. The OR2-OR1 sequence of phage λ was inserted into PEF-1α about 200 bp before TATA box. The artificial transcription activators can bind to OR2-OR1 sequence, and are thus conscripted near to the TATA box of PCMV-IE, activating gene transcription with artificial activating domain AH or VP2. All the components mentioned above were integrated into pED5, producing two vectors: pER-AH and pER-VP2. The two vectors were tested for their efficiency of expressing IFN-β and SEAP reporter genes in comparison with pERFRT, a vector similar to pER-AH and pER-VP2 except for lacking of artificial transcription activators. To abolish transcription efficiency affected by copy number or integration site in chromosome, all the vectors were integrated into FRT site by co-transfecting with an Flp recombinase producing plasmid. The FRT site was pre-inserted into CHO cell chromosome by Invitrogen Corporation. It was showed that pER-AH was only a little more efficient than pERFRT, however, pER-VP2 was 2.7 times more efficient than pERFRT in expressing reporter genes.]]> LAI Da-Zhi,WENG Shao-Jie,YU Chang-Ming,Qi Lian-Quan,FU Ling,YU Ting and CHEN Wei 1424true TANG Chao-Ke,YI Guang-Hui,WANG Zuo,WANG Yan,LIU Lu-Shan,WAN Zai-Yang,YUAN Zhong-Hua,RUAN Zhang-Geng and YANG Yong-Zong 1423true in vitro for a long time in addition to its hemagglutination property. This new lectin FRIL (Flt3-rectptor interacting lectin) was purified from Dolichos lablab, its biological speciality was identified and its receptors were located on hematopoietic stem/progenitor cells. When CD34+ cells were cultured in the presence of this lectin or its counterpart FL, within the 28 days of culture, FRIL can keep more cells in G0/G1 phase (beyond 80%) and 1.5 fold colony forming cells more than FL does after 14-day's culture. So, it is concluded that FRIL maintain HSC/HPCs self-renewal property by halting its cell cycles.]]> XIE Xiao-Yan,XIE Chao,Li Jin,Bai Ci-Xian and Pei Xue-Tao 1422true in vitro and it potentiates the transactivation properties of GR.]]> LI Shu-Rong,SU Yong-Ping,LIU Yun-Jie,LIU Xiao-Hong,LOU Shu-Fen and CHENG Tian-Min 1421true Escherichia coli. A total of 64 oligonucleotides were assembled in a single-step, in which T4 DNA ligase ligated DNA fragments from a pool of overlapping oligonucleotides. The synthetic gene was cloned to pSK vector. After sequencing and remending, the gene was expressed using the expression vector pET-28a-c(+). A band of 47 ku was detected with Western blot analysis. The protein so called MproteinAG has been covalently conjugated to the carboxyl-terminated magnetic particles. With the MproteinAG magnetic particles,IgG from blood serium of rabbit, pig,dog,mouse, human, macaque, rat, cat, sheep, cattle, horse can be purificated in one hour.]]> MENG Zhao-Hui,LIN Bing,XIE Yue-Hui and ZHANG Ke-Qin 1420true XING Li-Jing,ZHONG Kang,XU Zhi-Hong,WANG Rui and TAN Ke-Hui 1419true asd gene in an avirulent strain of S.flexneri to express coli surface antigen 6 of enterotoxigenic Escherichia coli. The results of Western-blotting demonstrated that avirulant strain of S.flexneri FWL01 expressed CS6 steadily. Immunofluorescence analysis showed that S.flexneri FWL01 carrying the plasmid pZLG6 can be excited fluorescence on its surface. Antibodies against CS6 and LPS of Shigella can be detected in sera of mice immunized with recombinant bacteria either orogastrically (o.g.) or intranasally(i.n.); simultaneously sIgA against CS6 can also be detected in the intestine. This is helpful for constructing multivalent recombinant vaccine for prevention of bacterial diarrhea.]]> LUO Gang,LI Shu-Qin,WANG Ling-Chun,ZHAO Ying,ZHENG Ji-Ping,DUAN Hai-Qing and ZHANG Zhao-Shan 1418true WANG Guo-Hua,Lü Jun-Hong,LEI Xiao-Ling,LI Hai-Kuo,LI Min-Qian,CHEN Run-Sheng,FANG Hai-Ping and HU Jun 1417true HE Hong-Xuan,QIN Xi-Ming,ZHANG Qiang-Zhe,TANG Yi,LIU Li-Yan,ZHENG Chang-Xue and DUAN Ming-Xing 1416true LI Zheng,CHEN Xi,TENG Hu,XIU Zhi-Long,SUN Li-Hua and FENG En-Min 1415true LIU Yun,WANG Cun-Xin,WANG Bao-Han and CHEN Wei-Zu 1414true cis-elements to bind the factor and alter gene expression by competition with AP-1 element.This decoy oligonucleotide restrained tumor cell proliferation in vitro and in vivo, and has important cancer gene therapeutic potential.]]> WANG Xue-Gen,YE Qing,CHEN Wu-Ling,FENG Ying,WEI Ce,LIU Hui and OUYANG Ping-Kai 1413true YE Zhong-De,SHEN Bei-Fen and LI Yan 1412true in vitro was established for high throughout screening. The MC4R receptor plasmid (MC4R/pCDNA3.1) and reporter gene plasmid (3×CRE/3×MRE/SRE-LUC) were cotransfected HEK293 to establish a cell line for agonist screening. To identify and optimize the assay condition, the effects of some factors were examined by using α-MSH, such as cell number per well, incubation time, final concentration of DMSO and luciferase's substrate concentration on the assay. A steady cell line and a reliable method were established for MC4R agonist screening, the Z'-factor value was near to 0.7 on the condition in each well that the cell number were 4×104 , the agonist incubation time was 8 h, the final concentration of DMSO was less than 1%, and the final concentration of α-MSH was 1 μmol/L. This technology can be applied to identify agonist for MC4R receptor.]]> ZHENG Xu-Xu,OUYANG Ke-Qing,GAO Hong,XU Zhi-Liang,HU Ying-He and CAI Shao-Xi 1411true ZOU Zhi-Qing,ZHOU Tian,ZHAO Jian-Long and XU Yuan-Sen 1410true Archaeoglobus fulgidus Na+/H+ antiporter (napA-2), which is partially homologous with the peptide coded by mdnr411 mRNA. All these results show that a cDNA of a new DNA related gene which function is fully unknown have screened and cloned. The foundation for further study of the function of the new gene, and its possible role in the pathogenesis of mice diabetic nephropathy has been laid, which is valuable to further understand the mechanism of diabetic nephropathy.]]> ZHENG Jing-Min,LIU Zhi-Hong,ZHANG Xin,YUN Shi-Feng and LI Lei-Shi 1409true FAN Heng-Yu,MENG Xiao-Qian,HUO Li-Jun,TONG Chao,CHEN Da-Yuan and SUN Qing-Yuan 1408true Escherichia coli DH5α in the form of inclusion body.The target protein was refolded in vitro and purified. After lysis with trypsin and carboxypeptidase B, an insulin analogue was purified by DEAE ion exchange and RP-HPLC.The product was characterized by MALDI-TOF MS.Circular dichroism spectrum was studied. The results implied the changes of the secondary structure and the association.The association of the analogue was further studied by size exclusive chromatography with a strong monomeric property. The biology activities including RIA, RBA were assayed, exhibiting 63.5% and 114.4% those of native insulin respectively. The result of mouse convulsion test indicated the insulin analogue had in vivo activity nearly the same as native insulin.]]> HUAI Wen-Hui,YUAN Hua,ZHAO Rui and RU Bing-Gen 1407true Salmonella typhimurium strain expressing conservative region(AB) of Helicobacter pylori(Hp) adhesion X4072(pYA248-AB) in mouse model infected with Hp, the feasibility of X4072(pYA248-AB) in Hp vaccination was determined. Hp infected mouse model was established and applied in oral vaccination. The density of bacterial colonization was determined by the semi-quantitative bacterial culture assay. The T cell subsets of spleenocytes were assayed by flow cytometry (FCM) after immunization. IL-2 and IL-4 were detected through MTT. Mice sera and intestinal fluid were separatively tested for AB-specific IgG and IgA by ELISA. The results showed that after immune treatment, the eradicate rate of vaccine treatment group was 53.3%, which was significantly higher than that of other two control groups (X4072(pYA248-AB) vs X4072(pYA248) and PBS, P＜0.01). The Hp colonized density of vaccination group with Hp infection was significantly lower than that of other two groups (X4072(pYA248-AB) vs X4072(pYA248) and PBS, P＜0.01). Comparing the ratio of spleen CD4+/CD8+ T lymphocytic cells of the immuned mice by flow cytometer, it was found that the ratio of vaccine treatment group and mice typhoid fever germ control group were significantly higher than that of negative control group (X4072(pYA248-AB) and X4072(pYA248) vs PBS, P＜0.05) respectively. In addition, the ratio of vaccine treatment was significantly higher than that of mice typhoid fever germ control group (X4072(pYA248-AB) vs X4072(pYA248), P＜0.05). The further analysis of cytokine it was found that IL-2 and IL-4 of vaccine treatment group and mice typhoid fever germ control group were significantly higher than those of negative control group (X4072(pYA248-AB) and X4072(pYA248) and PBS, P＜0.05) respectively, meanwhile, the IL-4 of vaccine treatment group was significantly higher than that of mice typhoid fever germ control group (X4072(pYA248) vs PBS, P＜0.05). Humoral immunity analysis showed that IgA was only detected in the immune treatment group, and the IgG antibody titer of immune treatment group increased significantly than that of the other two group, and the titer reached 1∶10 000 two weeks after the second intensive immunization. By the animal experiments it is suggested that X4072（pYA248-AB）can eradicate or significantly reduce the Hp colonization on the mice stomach. The potential immune therapeutic mechanism included the increase of CD4+/CD8+ ratio, induction of Th1 and Th2 response, and production of the specific antibodies.]]> BAI Yang,CHEN Ye,WANG Ji-De,CHANG Shao-Hong,ZHANG Zhao-Shan,ZHOU Dian-Yuan and ZHANG Ya-Li 1406true TAN Yin-Fei,SUN Xiao-Yang,TANG Shuang,PIAO Yun-Shang and WANG Yan-Ling 1405true C.elegans, A.thaliana, D.melanogaster and human. The prediction sensitivity and specificity on the exon basis are 83.7%～94.5% and 87.8%～97.1% respectively for these genomes. Non-canonical splicing has also been analyzed. The prediction capacity of the present method is comparable with GeneSplicer and other current splice site detectors.]]> ZHANG Li-Rong and LUO Liao-Fu 1404true in situ detection. Various tissues were tested to find out those which give rise to strong backward SHG. Then backward SHG imaging was applied in the studies of human skins. The SHG intensity from gangrenosis skins was much lower than that from normal skins. The skins of diabetic patients, which showed no macroscopic abnormality, also displayed a distinct decrease in SHG intensity. The result indicated that collagen in these skins may have suffered from micro-structural disorder or other changes. SHG imaging may serve as a new diagnosis method for dermopathy or other diseases.]]> Lin Xing-Sun,Pan Lin,Hu Jin-Yun,Ma Hui and Chen Die-Yan 1403true In vitro studies have revealed that treatment of various human epithelial cancer cell lines with specific COX-2 inhibitors induce apoptotic cell death. However, little is known about tumor cell lines of hematopoietic origin. The effects of COX-2 specitic inhibitor-celecoxib on proliferation and apoptosis on K562 cells are sought to be determined. K562 cells were exposed to increasing concentration of celecoxib (0, 10, 20, 40, 80, 160 μmol/L). The growth inhibition was evaluated by trypan blue dye exclusion, MTT assay and colony- formation inhibiting test. Apoptosis was determined by DNA ladder in agarose gel. The morphology apoptotic cells was identified by AO/EB staining combined with fluorescence microscopy observation, and by caspase-3 assay. The percentage of apoptotic cells was determined by flow cytometry. A blocking experiment for caspase-3 was carried out by incubating K562 cells together with DEVD-fmk. In addition, whether there is an expression of COX-2 mRNA or protein in K562 cells were investigated by RT-PCR and Western blot. Celecoxib(40～160 μmol/L) can effectively inhibit the proliferation and induce apoptosis of K562 cells in a dose-dependent manner. IC50 of celecoxib for inhibiting cells proliferation was 46 μmol/L. The regulation of apoptosis induction was related to an upregulated caspase-3 expression and activation. When caspase-3 activity was blocked by DEVD-fmk, the effect of celecoxib on apoptosis induction was partly abolished. Importantly, It is first demonstrated that K562 cells indeed exists in COX-2 expression both mRNA and protein (IL-1β inducement assay and RT-PCR/Western blot confirmation), celecoxib (80～160 μmol/L) could significantly down-regulate the expression of COX-2 mRNA and protein. In conclusion, these data indicated that a selective COX-2 inhibitor celelcoxib inhibits cell proliferation, induces apoptosis in human K562 cells, thus suggesting that COX-2 inhibitors may be a promising strategy in treatment of CML.]]> ZHANG Guang-Sen and LIU Ding-Sheng 1402true ZHAO Yu,LUO Tian-Hong,WANG Kan-Kan,ZHANG Ji,ZHENG Pei-Zheng,ZHAO Chun-Jun,LIU You-Ping,ZHENG Sheng,GU Yan-Yun,LI Guo and LUO Min 1401true XIA Yun,LIN Ru-Xian,ZHENG Su-Jun,ZHANG Min-Li and WANG Sheng-Qi 1400true WANG Yang,ZOU Li-Juan,XU Guan-Dong,GONG Lin-Lin,ZHOU Xiao-Mian and SHAO Shu-Juan 1399true LIU Qiang,XU Jun and CHEN Run-Sheng 1398true HAN Ya-Ling,YAN Cheng-Hui,LIU Hai-Wei,HU Ye,KANG Jian,WANG Xiao-Zeng and LI Shao-Hua 1397true ZHOU Jin-Yong,HU Jing-Jing,ZHONG Yan,LIU Chao,CAI Wei-Dong,YUAN Li,TENG Li-Ping and DE Wei 1396true Arabidopsis thaliana AtNHX2 gene is one member of the Arabidopsis NHX Na+/H+ antiporter gene family and play an important role in salt tolerance. A sequence-analyzed 2.8 kb DNA fragment on upstream of ATG start codon of AtNHX2 gene was cloned into pCAMBIA1301-1. The promoter activity was detected by transient expression in onion epidermis. The reconstructed vector pCAMBIA1301-1/ AtNHX2 promoter was transformed into Arabidopsis thaliana by Floral Dip method. AtNHX2 promoter-GUS analysis in transgenic Arabidopsis showed that AtNHX2 was expressed in all tissues. Strong GUS expression was detected in guard cells suggesting the possibility that it is involved in stomatal regulation. AtNHX2 promoter activity was decreased by NaCl and up-regulated by KCl, demonstrating that NaCl and KCl regulation to AtNHX2 expression occurs at the transcriptional level. GUS activity in old leaves was higher than in new leaves, which reveals a role of AtNHX2 in salt tolerance. Strong GUS activity in root hair cells was observed. It suggests that AtNHX2 may play an important role in partitioning of Na+ into the enlarged vacuoles of root hair cells.]]> LI Jin-Yao,XU Li,MA Ji,ZHOU Jie and ZHANG Fu-Chun 1395true YANG Xiang-Dong,WANG Ren,LIU Jun-Wen,QU Shun-Lin,GUO Fang,WANG Lei and SHENG Shu-Li 1394true Pichia pastoris strain GS115. The recombinant proteins, HRGD1 and HRGD2, were expressed and induced by methyl alcohol, secreted into media leading by signal peptides and purificated by combination of ultrafiltration, cation exchange chromatography and gel filtration chromatography. The biological activity assays approved the chimera proteins show activity not only in antithrombin but also in antiplatete aggregation as what have been predicted.]]> ZHAO Jing-Jing,YANG Jiang-Feng and RU Bing-Gen 1393true WANG Li-Yue,DIAO Lu-Ming,TIAN Qing,WANG Jian-Zhi and GONG Cheng-Xin 1392true in vivo angiostatic effects on chick chorioallantoic membrane(CAM) by recombinant mouse canstatin N-fragment(1～95aa), the cDNA for N-fragment of mouse canstatin, obtained from a cloning vector pMD18T-mCan by PCR, was introduced into an expression vector pET30a(+) to construct prokaryotic expression vector pET-mCanN. N-fragment of mouse canstatin efficiently expressed in E.coli BL21(DE3) after IPTG induction was monitored by SDS-PAGE and by Western blotting with an anti-hexahistidine tag antibody. The expressed N-fragment of mouse canstatin, mainly as inclusion bodies, accounted for approximately 18% of the total bacterial proteins as estimated by densitometry. The inclusion bodies were washed, lysed and purified by the nickel affinity chromatography to a purity of 92%. The refolded N-fragment of mouse canstatin was tested on the chicken embryo CAMs, and a large number of newly formed blood vessels were significantly regressed. It is concluded that N-fragment derived from mouse canstatin is an effective inhibitor of angiogenesis of the chicken embryo in a dose-dependent manner through CAM assay and a promising candidate for the treatment of cancer, suggesting that these novel findings may add to the understanding of anti-angiogenesis effects of the N-fragment of mouse canstatin.]]> HOU Wei-Hong,JIA Yan-Long,WANG Tian-Yun,CHAI Yu-Rong,YUAN Bao-Mei,TIAN Fang,WANG Jian-Min and XUE Le-Xun 1391true 6 cells/ml and 1×106 cells/ml onto scaffolds respectively to culture for 2 weeks. The biological properties of the fabricated bone tissue were detected by inverted microscope, scanning electron microscope (SEM), alkaline phosphatase (ALP), von-Kossa staining on mineralized nodules and AO/EB fluorescence staining. Meanwhile the cells' metabolism of nutrients was monitored and analyzed during the whole culture process. The bone tissue fabricated in RWV with two different seeded cell densities grew well. Osteoblasts in the scaffolds secreted much collagen fibers and formed mineralized nodules and new osteoid tissue. It can be conclueled that: With the stress stimulation inside the fluid in the RWV, the active expression of ALP can be increased, the formation of mineralized nodules can be accelerated. The rapid proliferation and differentiation of osteoblasts are possible and the three-dimensional fabrication of engineered bone could be realized.]]> SONG Ke-Dong,LIU Tian-Qing,LI Xiang-Qin,CUI Zhan-Feng,GE Dan,SUN Xiang-Yu and MA Xue-Hu 1390true WU Xiao-Ying,CHEN Zhu-Chu,XIAO Zhi-Qiang,LI Cui,WANG Chun-Nian,LI Jian-Ling,FENG Xue-Ping and ZHANG Peng-Fei 1389true Deinococcus radiodurans sequenced genome, which has been found in GenBank database through amino homology blast base on combination of bioinformatics. Blasting against the database indicates that amino acid sequence of this ORF has less than 60% homology with the reported terhalose synthases. The function, which was identified by expressing this ORF in Escherichia coli, proves this ORF is a novel trehalose synthase gene. Using the 30% maltose as substrate, about 65% maltose can be converted into trehalose by this enzyme. Recombinant enzyme showed optimal activity at pH 7.0 and 30℃ when it converts maltose into trehalose.]]> WEI Yu-Tuo,ZHU Qi-Xia,LUO Zhao-Fei,CHEN Fa-Zhong,LI Gui-Yuan,HUANG Kun and HUANG Ri-Bo 1388true LI Jing,SONG Guang-Yao,JIANG Ling-Ling,ZHANG Wen-Jie and FENG Zhi-Shan 1387true DPYD gene, is the major rate-limiting enzyme in the metabolism of the anti-cancer drugs. And it exists substantial inter-individual variations in its enzymatic activity. It was shown that mutant alleles of DPYD that encoding an inactive/decreased enzyme are largely caused by SNP (single nucleotide polymorphism) in the gene. Detection of the SNPs is the basis for the prediction of drug-response and realization of individualized therapeutic regimen for patients. For this reason an oligonucleotide microarray for genotyping 6 known SNPs of DPYD gene was optimized, and the genotyping standard for this microarray was fabricated. The microarray system was used to detect the mutant allelic frequency of DPYD in cancer patients (112 individuals), nephrotic patients (83 individuals) and healthy subjects(45 individuals). In the studied groups, the average frequencies for the mutant alleles of DPYD*5 and DPYD*9 are 11.25% and 32.08% respectively. No mutant alleles of DPYD*2, *3, *4, *12 were found. And there is no significant difference (P＞0.05) in the mutation incidence in different subject groups or sex groups. The SNPs was shown to have no significant association with the disease and sex. The genotyping results of 20 samples were tested by direct sequencing, the genotyping results of 19 samples by miroarray were conformed with the direct sequencing. The allelic frequencies of DPYD*5 and DPYD*9 is high in the studied groups, quick and accurate detection of them can be achieved by using DNA microarray.]]> WEN Si-Yuan,WANG Xiao-Yun,ZHANG Min-Li and WANG Sheng-Qi 1386true E.coli BL21. The results of gel scanning and Bradford protein assay show that the BmVUB protein was about 9 ku and the yield consists of 50 percent of total bacterial protein. The concentration of purified BmVUB protein was 1.03～2.46 g/L. The high titer antibody was obtained from rabbits immunized with the (His)6- BmVUB fusion protein and ELISA shows that the titer of antibody was over 3.2×10-5. Peptides binding ubiquitin were obtained from phage displayed peptide library using BmNPV-ZJ ubiquitin purified with His-affinity chromatography as a ligand. Five short peptides were identified which displayed obvious biological activity. Peptide VAPHHAYAPMRT could regulate the cell proliferation and further more this function which depends on the concentration variation, namely low concentration could promote the cell proliferation, whereas high concentration will inhibit the proliferation.]]> ZHANG Wen-Bo,LIN Rong,ZHANG Yao-Zhou and WU Xiang-Fu 1385true Xenopus oocytes, green fluorescence of the fusion protein was only found at the cellular membrane. To further determine subcellular localization signal of SDCT2, its N-terminal and C-terminal sequences were deleted and fused with EGFP. These deletion mutants were transfected into LLC-PK1 and their intracellular distributions were observed. The confocal analysis indicated that the SDCT2 with N-terminal deletion was predominantly distributed in the cytoplasm and also expressed at apical and basolateral membranes. The SDCT2 with C-terminal deletion was almost entirely distributed at the basolateral membrane, nearly not expressed at the apical membrane, and seldom expressed in the cytoplasm. Immunohistochemistry staining revealed that SDCT2 only expressed at the basolateral membrane of proximal renal tubular cells. The above results suggested that the N-terminus of SDCT2 is required for its subcellular localization and the basolateral membrane localization signal of SDCT2 locates in the N-terminal sequence.]]> BAI Xue-Yuan,HOU Kai,CHEN Xiang-Mei,FENG Zhe,FU Bo and WU Di 1384true 2 liver cells were exposed to 9 mmol/L alloxan for 1 h and then incubated in fresh culture media for 0, 2, 8 and 24 h respectively. The frequencies of oxidative base damage in mtDNA were measured by a quantitative Southern blot coupled with digestion by the enzymes endonuclease Ⅲ and formamidopyrimiding DNA glycosylase. Next, ligation-mediated PCR (LM-PCR) was performed to map damage to specific nucleotides along a ～100 bp fragment including MTTL1 gene. The addition of alloxan to cultured human cells increased the rate of oxidative base damage and ,by several fold, the lesion frequency in mtDNA. After removal of this DNA damaging agent from culture, the lesion frequency decreased to levels slightly higher than normal at 8h and returned to normal levels at 24 h. The result of LM-PCR showed 20 hot spots of MTTL1 gene where nucleotides were receiving a majority of damage. The pattern of oxidative damage is like that of point mutation identified in this gene, suggesting the possibility that oxidative damage mainly contributes to the formation of the point mutation in MTTL1 gene.]]> WANG Zhen-Cheng,WANG Xue-Min,MIAO Ming-Yong and JIAO Bing-Hua 1383true XIAO He,YU Ming,LI Song,SHEN Bei-Fen and LI Yan 1382true Mycoplasma hyorhinis in gastric tumor tissues suggests a possible association between mycoplasma infection and cancer. P37, an outer-membrane bacterial protein from Mycoplasma hyorhinis, increases neoplastic invasivity and metastasis and induces TNF-α secretion from human peripheral blood mononuclear cells (PBMC). To investigate the functional mechanism of P37, yeast two-hybrid system was used to isolate proteins interacted with P37 from a human placenta cDNA library. Among the 2.6×106 transformant clones, one positive clone was obtained. Sequence analysis revealed that the clone is a cDNA fragment from Norpeg and encode a carboxy terminal protein. The interaction between P37 and Norpeg was further confirmed by ELISA and GST-Pull down assay. This result would be useful to study the function of P37 further.]]> SUN Yu-Ning,JIN Geng-Lin,ZHANG Jian-Zhi,WU Jian and SHOU Cheng-Chao 1381true E.coli in large amounts, however this may result in the formation of inclusion bodies, and the recovery of biologically active products becomes significant. As the minimal mechanism of GroEL-mediated protein folding, minichaperones, fragments encompassing the apical domain of GroEL, can facilitate the refolding of several proteins in vitro without requiring GroES, ATP, or the cage-like structure of multimeric GroEL. Here minichaperone (GroEL191～345)-mediated in vitro refolding of recombinant human interferon gamma (rhIFN-γ) in free and immobilized GroEL191～345 systems are studied. SDS-PAGE was performed with Laemmli's Tris-glycine buffer system for protein analysis. Protein concentrations were determined by using a modification of Lowrys method with bovine serum albumin as reference and total activities of renatured rhIFN-γ were measured by CPE method. The results showed that the presence of GroEL191～345 in the refolding buffer not only enhanced the specific activity, but also promoted the refolding efficiency. With the initial protein concentration of 100 mg/L, the protein yield and total activity of rhIFN-γ assisted by GroEL191～345 was 2.2 and 3 folds of that under spontaneous condition respectively. Optimal operating parameters in refolding of rhIFN-γ assisted by GroEL191～345 were as follows: refolding temperature 15℃, refolding time 4 h, pH 7.7, initial concentration of IFN-γ 100～200 mg/L and the molar ratio of GroEL 191～345 versus IFN-γ 1∶1～2∶1. Furthermore, the immobilization of GroEL191～345 on NHS-activated sepharose fast flow made it possible to be recycled. The protein yield and the specific activity of rhIFN-γ were 46.29% and 1.95×107 U/mg respectively even the initial protein concentration was up to 400 mg/L.]]> GUAN Yi-Xin,FEI Zheng-Zheng,LUO Man and YAO Shan-Jing 1380true LI Shu-Tian,TIAN Ying-Chuan and HE Chao-Zu 1379true QU Mei-Hua,LI Hui,XU Yan-Juan and HE Rong-Qiao 1378true in vitro and the expression of survivin was analyzed by RT-PCR, Western blotting and cell cycle. There were significant transcription of survivin and detectable protein of survivin in PBMC cultured with PHA and IL-2 for 36 h. The expression of survivin increased with the extension of culture time and this expression was cell cycle dependent. AG490 (JAK2 inhibitor) resulted in the cell cycle arrested and inhibited the expression of survivin markedly, and Wortmannin(PI3K inhibitor) and PD98059(MEK inhibitor) also inhibited the expression of survivin to a less extent. The results revealed that the expression of survivin correlated with the activation of peripheral lymphocyte and meant the state of cell proliferation. The JAK-STAT signal transduction pathway mediated by JAK2 was essential for the activation of peripheral lymphocyte and the expression of survivin. And the signal transduction pathway mediated by PI3K and MEK also affected this process to a certain extent.]]> DONG Yan,MEI Zhu-Zhong,SONG Yi,SUN Guo-Jing,QIAN Jun-Jie,LIU Bin,LIU Hua and SUN Zhi-Xian 1377true 3H］LDL. During the observation, the amount of Ox-r［CL-3H］LDL taking up increased gradually in both groups, however, from 24 h the taking up amount in antisense treated cells was less than that of untreated control. There was a statistical difference between the two groups from 24 h to 96 h. The activity of ACAT in the two groups was observed as well. From 6 h to 48 h, the relative ACAT activity increased in both groups. The relative ACAT activity kept unchanged from 48 h to 96 h in the two groups. At 48 h, the relative ACAT activity in antisense group was significantly lower than that of control group. The activity of ACAT was related to adipophilin expression but was not a linear relationship. This meant, at least partly, expression of adipophilin was related to cellular taking up of lipoproteins and to ACAT activity. Therefore, it is indicated that adipophilin has a role in metabolism of cellular lipid droplets and adipophilin is associated with ACAT.]]> YUAN Zhong-Hua,YANG Yong-Zong,YIN Wei-Dong,YI Guang-Hui and WAN Zai-Yang 1376true BAI Bing-Xue,XU Dong-Gang,WANG Jin-Feng,PENG Shan-Yun,ZOU Min-Ji and FAN Ming 1375true P＞0.05), and the protein and mRNA level of PPAR-γ in ventricles of 4-week-old SHR were also similar to that of age matched WKY (P＞0.05). In 16-week-old SHR, the ratio of LVWM and ISWM to BM were higher compared with age matched WKY (P＜0.01), whereas there was no significant difference in RV (P＞0.05). The protein and mRNA level in LV and IS of 16-week-old SHR were great lower than that of 16-week-old WKY (P＜0.01), while in RV there was no significant difference of protein (P＞0.05), and a little decrease of mRNA (P＜0.05). The findings suggested that the expression level of PPAR-γ in hypertrophied ventricles induced by pressure overload (LV and IS in 16-week-old SHR) was significantly decreased, and the decreased expression of PPAR-γ in ventricles might contribute to ventricular remodeling in hypertension.]]> QI Ying-Xin,NIU Xiao-Lin and ZHAO Zhong-Liang 1374true FENG Jin-Li,HU De-Hui,CHEN Ming,TIAN Ying-Hong,SHI Zhi-Min,YANG Jian-Ming,ZHU Xin-Hong,LI Xiao-Wen and GAO Tian-Ming 1373true JIA Yun-Li,YU Ze-Hua and CHEN Xin-Wen 1372true SHI Wei,HAN Shu,LI Yan-Hua,XIE Xiao-Yan,SHI Shuang-Shuang,CHEN Lin,BAI Ci-Xian,YAN Fan,NAN Xue,WANG Yun-Fang and PEI Xue-Tao 1371true PENG Cong,LI Xiao-Ling,ZHOU Ming,LIU Hua-Ying,WANG Li-Li,ZHANG Qiu-Hong,YANG Yi-Xin,WU Shang-Hui,HUANG Bo-Ying,XIONG Wei and LI Gui-Yuan 1370true JIANG Lin,FENG Jing,WU Wen-Fang,YANG Dong-Ling and YAN Xi-Yun 1369true LI Min,Chen Li-Mei,LIN Ai-Xing,YANG Xing-Yuan,AN Xiao Rong and CHEN Yong-Fu 1368true HU Zhong-Liang,WEN Ji-Fang,XIAO De-Sheng,ZHENG Hui and FU Chun-Yan 1367true FENG Zhe,CHEN Shang-Bin,LI Peng-Cheng,ZENG Shao-Qun and LUO Qing-Ming 1366true QI Bi-Shu,LI Chun-Guang,CHEN Ye-Miao,LU Ping-Li,HAO Fu-Shun,SHEN Guo-Ming,CHEN Jia and WANG Xue-Chen 1365true LIU Chang-Zhen,HE Li,SHEN Qing-Tao,LI Xiang-Hui and SUI Sen-Fang 1364true ZHANG Qiang-Zhe,QIN Xi-Ming,DONG Hai-Li,LIANG Rong,HE Hong-Xuan,LI Xi,JIANG Bei-Yu,LIU Xiang-Jun and DUAN Ming-Xing 1363true LIU Yuan-Yuan,ZHAO Ping,WU Zheng-Xing,QU An-Lian and XU Tao 1362true LIU Chun,ZHAO Ping,CHENG Ting-Cai,ZHA Xing-Fu,XIA Qing-You and XIANG Zhong-Huai 1361true WEN Si-Yuan,CAO Heng-Jie,LIU Jun-Bo,DING Yu,CHEN Su-Hong and WANG Sheng-Qi 1360true ZHU Cheng-Gang and DING Zhi-Shan 1359true MA Wen-Li,LIU Jia,LI Ling,ZHANG Bao,SHI Yong-Xia and ZHENG Wen-Ling 1358true YU Da-Hai,CAI Hong and ZHU Yu-Xian 1357true ZHANG Xiao-Yong,ZHANG Wei,GAO Ping,CHANG Zhi-Jie,SUN YI-NA and LIU Hui-Tu 1356true NIE Xin-Min,GUI Rong,LI Deng-Qing,ZHOU Ming,HUANG Zu-Fa and LI Gui-Yuan 1355true LI Qing-Hua,PANG Tian-Xiang and HAN Zhong-Chao 1354true WANG Li,WANG Xiao-Chuan,LI Hong-Lian,WANG Dan-Lin,ZHOU Xin-Wen and WANG Jian-Zhi 1353true WANG Li-Li,ZHANG Qiu-Hong,MA Jian,PENG Cong,CAO Li,TANG Ke,LI Xiao-Ling and LI Gui-Yuan 1352true LI Xiao-Jing,ZHANG Hao,XUE Chong,LI Yan-Ying,CHEN Jing,MIAO Lin,FANG Hong-Qing and CHEN Hui-Peng 1351true LI Hong-Mei,MEI Le-He,URLACHER VLADA and SCHMID ROLF D 1350true WANG Fang-Yu,ZHANG Tian-Yuan,LUO Jin-Xian,LI Rui-Fang,GAN Jing-Jing,GUAN Wen-Jun and XIAO Fan 1349true SHI Bi-Zhi,GENG Fei,HU Ping,HE Pei-Jie and WU Xing-Zhong 1348true YANG Xiao-Dong and LIU Jin-Yuan 1347true ZHAO Jian,XU Jun-Jie,MIAO Jing,LI Bing,YANG Xiu-Xu,SONG Xiao-Hong and CHEN Wei 1346true HUANG Biao,TAO Wen-Yi,ZHANG Lian-Fen,SHI Jin,YANG Hai-Lin and JIN Jian 1345true ZHAO Hui,WANG Wei,ZHANG Qing-Run,GAO Yang,ZHAO Hong-Bin,ZHOU Jun,LIN Wei and ZENG Chang-Qing 1344true FENG Ju-Tao,LIU Yin-Kun,MOHAMAD RADWAN ALMOFTI,DAI Zhi,ZHOU Hai-Jun,SONG Hai-Yan,LU Hao-Jie and ANG Zhao-You 1343true LIU Yu,HE Li-Qiang,TAN Zhi-Ping,TANG Cheng-Yuan,CHI Jing-Wei,CAI Fang,PAN Qian,LONG Zhi-Gao,LIANG De-Sheng,WU Ling-Qian,DAI He-Ping,XIA Kun,ZHANG Zhuo-Hua and XIA Jia-Hui 1342true ZHANG Xiao-Peng,XIAO Zhi-Qiang,LI Cui,LI Jian-Ling,YU Yan-Hui,OYANG Yong-Mei,FENG Xue-Ping,ZHANG Peng-Fei and CHEN Zhu-Chu 1341true 2( ＋ )/CREG and pLXSN( － )/CREG, were constructed. Western blot and immunoflourescence analysis showed that the expression of CREG and SM α -actin increased in HITASY after infection with pLNCX2( ＋ ) /CREG. The migration of HITASY infected with pLNCX2( ＋ )/CREG obviously enhanced compared with that of normal HITASY and pLXSN( － )/CREG cells showed by scrape-wounding and time-lapse analysis. Moreover, CREG over-expression increased the secretion of MMPs in HITASY tested by Western blot. Gelatin SDS-PAGE zymography analysis revealed that the activities of MMPs also increased in HITASY infected with pLNCX2( ＋ )/CREG . On the other hand, the opposite effects were observed when CREG expression decreased by using antisense pLXSN( － )/CREG . These results suggest that CREG may be able to induce the VSMCs differetiation and promote the VSMCs migration in the meantime.]]> HAN Ya-Ling,HU Ye,LIU Hai-Wei,KANG Jian,YAN Cheng-Hui and LI Shao-Hua 1340true LIU Yan-Jun,LUO Hai-Bo,ZHU Ping and FU Ning 1339true E204Q and tri-mutant BRI119T/T121S/A126T of bacteriorhodpsin were constructed by directed-mutation method. The M412 Flash-dynamics spectrum indicates that the life of M intermediate of the single mutant BRE204Q is 7.10 ms and the life of M intermediate of the tri-mutant BRI119T/T121S/A126T is 8.23 ms, whose lives of M intermediate are longer than that of the wild-type BR with life of the M intermediate 6.23 ms. The result shows that the tri-mutant has the longest M intermediate life which exceeds wild-type BR by thirty-two percent. The Proton-pump functional research of the two mutants illustrates that their proton-pump functions decresae compare with that of wild-type BR respectively and the proton-pump function of the tri-mutant BRI119T/T121S/A126T is the weakest one.]]> SONG Jing-Jiao,ZHONG Sheng,ZHANG Yue,CAO Jun-Wei and HU Kun-Sheng 1338true XU Rong,SHEN Yi-Jun,DENG Song-Hua,CAI Chun-Xiao,CHEN Qiu-Li,JIA Jian-An,JIA Jian-An,PAN Xin and PAN Wei 1337true 4S)3, and this fusion protein was a potential targeting thrombolytic agent. But the fusion protein leaned to be proteolysed while expressed in CHO cells. To overcome this problem, four new linkers were selected from linkers database by using the program at http://ibivu.cs.vu.nl /programs/ linkerdbwww, and four IIn-UK fusion genes were reconstructed by replacing linkers. And other four fusion genes were reconstructed by changing reletive position of two moities and replacing the linker. The degree of proteolysis of eight reconstructed IIn-linker-UK or UK-linker-IIn fusion proteins were analysed with Western blot by using anti-urokinase antibody, the results showed that all eight constructed fusion proteins were degraded partly while expressed in CHO cells. So three new IIn-linker UK fusion genes were constructed by using a new linker coming from pro-urokinase and removing one or two cleavage site of proteolytic enzyme around the linker. The Western blot showed that IIn-UK fusion protein removed two protyolitic enzyme cleavage sites had good anti-proteolysis ability in COS7 cell and CHO cell. Thus it laid a foundation for preparation of IIn-UK fusion protein in CHO cells and further research of targeting thrombolytic agent.]]> LIU Zhi-Gang,LIN Jian-Bo,Du Yun and Yu Wei-Yuan 1336true JIN Si,CHEN Jian-Guo,ZHU Li-Ping,LIU Sheng-Yuan,WANG Di-Xun and HU Qing-Hua 1335true XU Xia-Lian,XIN Hong,ZHANG Xin-Jun,REN Fang-Li,WANG Yin-Yin and CHANG Zhi-Jie 1334true LI Xin-Hui,JI Tong,SUN Jie-Lin,SUN Jie-Lin and HU Jun 1333true E.coli ML-35p (an ampiciline-resistance strain), Pseudomonas aeruginosa ATCC 27853, and Candida albicans ATCC 10231 were 12.5, 25, and 100 mg/L respectively. In contrast, the recombinant holo-HMG-17 was inactive against Staphylococcus aureus ATCC 25923. The immunocytochemistry staining, ELISA, and Western blotting revealed that HMGN2 was present in the cytoplasm of mononuclear leukocytes and released to the extracellular environment when stimulated with IL-2. The results indicated that HMGN2 was a new effector molecule of human LAK cells.]]> Feng Yun,Xiong Wen-Bi,Wang Guo-Xing,Huang Ning,Wu Qi,Bao Lang,Li Xuan and Wang Bo-Yao 1332true YANG Li-Ye and HUANG Tian-Hua 1331true LIU Mei-Ling,PENG Jing-Pian,SUN Quan-Hong,YANG Ying and XIA Hong-Fei 1330true YE Li-Hong,YOU Jia-Cong,QIAO Ling,CAI Na,WANG Chang-Ye,LIU Yi,WU Jing-Hui,WU Lian-Ying,WANG Hong-Hui and ZHANG Xiao-Dong 1329true Qiu Xue-Mei,Li Ning,Deng Xue-Mei,LIAN Zheng-Xing,Wang Qi-Gui,Wang Xiu-Li and Wu Chang-Xin 1328true YU Quan-You,LI Bin,LI Guan-Rong,FANG Shou-Min,YAN Hong,TONG Xiao-Ling,QIAN Ji-Feng,XIA Qing-You and LU Cheng 1327true DAI Zhi,LIU Yin-Kun,CUI Jie-Feng,FENG Ju-Tao,SONG Hai-Yan,CHEN Jie,SUN Rui-Xia,LI Na,SHEN Hua-Li,ZHANG Yu and YANG Peng-Yuan 1326true ZHANG Qian and WANG Jing-Ze 1325true WT (wild type), pcDNA3.1(-)-HSP70ΔNLS (truncated mutant of HSP70 without NLS), pEGFP-N1-HSP70WT, and pEGFP-N1- HSP70ΔNLS were constructed respectively. To mimic oxidative stress in vitro, final concentration of 1.0 mmol/L hydrogen peroxide (H2O2) was added into cultured C2C12 myogenic cells. Toluidine blue staining showed predominantly compact, centrally localized nucleoli in intact control cells, but in H2O2-treated cells, an early onset of nucleolar segregation could be found after 3 h. The nucleolar segregation could be inhibited significantly in heat shock-pretreated cells and pcDNA3.1(-)-HSP70WT-transfected cells. Fluorescence microscopy and immunoblotting for nucleolar protein showed that HSP70WT was localized within cytoplasm in normal cells and redistributed to nucleus and nucleolus at 1 h after treatment with H2O2. NLS deletion mutation of HSP70 (pcDNA3.1(-)-HSP70ΔNLS) abolished H2O2-mediated translocation of HSP70 from cytoplasm to nucleus and nucleolus, and subsequently, its inhibitory effect on H2O2-mediated nucleolar impairment. It was suggested that wild type of HSP70 can attenuate significantly nucleolar segregation induced by oxidative stress. By mediating translocation of HSP70 from cytoplasm to nucleus and nucleolus, NLS played important role in the inhibitory effect of HSP70 against nucleolar impairment induced by oxidative stress.]]> WANG Kang-Kai,E Shun-Mei,JIANG Lei,ZHANG Hua-Li,LIU Ke,ZHANG Ling-Li and XIAO Xian-Zhong 1324true NING Hong-Xiu,CHEN Yue,ZHANG Yuan-Jiang,RONG Yu,WU Xiao-Jun,ZHANG Xiu-Fang and CHANG Zhi-Jie 1323true ZHANG Qiu-Hong,WANG Li-Li,PENG Cong,CAO Li,WANG Jie-Ru,LI Xiao-Ling and LI Gui-Yuan 1322true CHEN Li-Yu,YI Xin-Yuan,ZENG Xian-Fang,CAI Chun and L.McREYNOLDS 1321true 2=10.214 ， P＜ 0.005). The over expressions of nm23-H1, DJ1 and TIM1 in both the NPC tissues and the cell line may be correlated with the pathogenesis of poor differential nasopharyngeal squamous carcinoma.]]> FENG Xue-Ping,CHEN Zhu-Chu,XIAO Zhi-Qiang,YANG Hai-Yan,ZHUGE Qin,ZHU Guo,YANG Yi-Xuan,LI Ming,LI Cui,LI Mao-Yu,LI Feng,ZHANG Peng-Fei and XIAO Jian-Yun 1320true M.tuberculosis DNA vaccine, DNA vaccines encoding Ag85B, MPT64, MPT70 and PstS-3 protein were constructed with eukaryotic expression vector pJW4303. Combination DNA vaccines were inoculated intramuscularly into C57BL/6 mice three times at 3 weeks interval. After 21 days of the third injection, the specific antibody titers against the four antigens were 1∶6 400, 1∶51 200, 1∶6 400 and 1∶6 400. Meanwhile the mixed spleen cells could produce the high antigen-specific IFN-γ level in response to the four antigen proteins, IFN-γ level of Ag85B, MPT64, MPT70 and PstS-3 reached 10 582.14 ng/L, 13 635.97 ng/L, 14 213.15 ng/L and 9 657.35 ng/L respectively. After the last injection, mice were challenged with M.tuberculosis H37Rv. When compared to negative group, the bacterial counts of lung and spleen from the mice vaccinated with tetravalent combination vaccine were reduced about 650 and 130 folds. Microphotographs showed clearly that lungs of mice vaccinated with combination vaccine were much better protected against Mycobacterium tuberculosis challenge than negative mice. The results showed that tetravalent combination M.tuberculosis DNA vaccine elicited both T-cell and humoral immune response, and protected against tuberculosis effectively.]]> GU Tian-Yuan,CAI Hong,TIAN Xia,YU Da-Hai and ZHU Yu-Xian 1319true LI Wen-Hai,DENG Xue-Mei,LI Ning,Wang Shao-Hua,ZHAO Yi-Qiang,ZHAO Yi-Qiang,ZHANG Ran,WU Ke-Liang and WU Chang-Xin 1318true in vivo cloning. Linear PCR fragments that amplified from low copy plasmid pACYC184 were used as gene targeting vector. The length of subcloned DNA sequence including Red gene and a series regulatory sequences are about 6.7 kb. The biology function of Red gene in pYM-Red was tested by gene replacement (galk<>kan) of W3110 chromosome. Factors that effect recombination efficiency were precisely confirmed. When induced 10 min at 42 ℃ and using 300 ng linear DNA fragment as targeting molecules, the efficiency of pYM-Red mediated recombination can reach one positive recombination clone per four thousands electroporation survived cells, it is 5~6 folds higher than pBR322-Red and pKD46 recombination system.]]> YU Mei,ZHOU Jian-Guang,CHEN Wei,LI Shan-Hu and HUANG Cui-Fen 1317true BAI Xiang-Yang,NI Jian-Feng,Lü An-Guo,WU Wen-Fang and NIU Rui-Fang 1316true SHI Xin-Hui,QIN Chung-Deng,SONG Jian-Lan,XIA Xiao-Ping and WANG He 1315true YANG Ming,FU Han-Jiang,TIE Yi,ZHU Jie,JIANG Hong and ZHENG Xiao-Fei 1314true TANG Chao-Ke,FENG Da-Ming,SUN Wen-Qing,LIU Lu-Shan,YI Guang-Hui,Yang Jun-Hao,WANG Zuo,Wang Shuang,WANG Zai-Yang,YANG Bao-Tang and YANG Yong-Zong 1313true BamHⅠ-XhoⅠ restriction sites of the GST fusion expression vector pGEX-6P-I, in which there is a rhinovirus 3C protease cleavage site for the fusion protein. E. coli strain BL21 (DE3) was transformed with the recombinant GST fusion plasmids. The supernatants lysed by sonication and clarified by centrifugation were passed over Glutathione-Sepharose 4B column for purifying, respectively. The GST fusion proteins were then cleaved by GST-fusion rhinovirus 3C protease and then were purified by the affinity chromatography. The LearnCoil-VMF and ExPASy bio-softwares were used for predict and analysis the structure and function of five peptides. And the results of Gel-filtration and circular dichroism (CD) showed that the HR1 and HR2 form a six-helix structure.]]> WANG Xiao-Jia,ZHU De-Bing,ZHANG Guo-Zhong,BAI Ya-Duo and WANG Ming 1312true P<0.01) decreased in hypertensive subjects. In vitro incubation with ( － )epicatechin caused an increase in GSH and — SH content, the effect was more pronounced in hypertensive erythrocytes. Similar results were obtained with L-ascorbic acid. The observed decrease in the level of GSH and — SH groups in hypertension is an indicator of oxidative stress condition. Observation of an increase in red cell GSH content and the protection of membrane — SH group oxidation by ( － )epicatechin in hypertensive subjects is a convincing reason to suggest that high dietary intake of foods rich in catechins may help to reduce oxidative stress and concomitant free radical damage in hypertensive patients.]]> SYED IBRAHIM RIZVI and NAVNEET KUMAR 1311true Pseudomonas aeruginosa is re-investigated with differential scanning calorimetry (DSC) and circular dichroism (CD) methods. The results show that unfolding in the pH range from 4 to 9 is associated with two heat capacity maxima. The low temperature transitions are reversible at all pH conditions used, while the high temperature transitions are irreversible. The two unfolding transitions were analyzed by the two-interchangeable-conformation model with the fraction for the first transition (N1) from 64% at pH 4.0 to 55% at pH 9.0. Temperature induced unfolding monitored at 219 nm shows also two separate transitions. The ratio of the signal changes is consistent with the fractions obtained from the corresponding DSC measurements. These results provide further support for the hypothesis that at least two conformations of apoazurin coexists in solution.]]> ZHANG Hong-Jie 1310true Kd values as low as 140 nmol/L. Aptamer H2 (10 μmol/L) inhibited approximately 44% of the helicase activity of HCV NS3 in vitro. The results suggest that aptamer against HCV NS3 helicase have been identified by means of SELEX methods from an 81bp single stranded DNA random library.]]> ZHAN Lin-Sheng,ZHUO Hai-Long,WANG Hui-Zhong,PENG Jian-Chun and WANG Quan-Li 1309true Torulopsis glabrata was studied by adding oxidative phosphorylation inhibitors (rotenone, antimycin A and oligomycin). When 10 mg/L rotenone and antimycin A were added to the cell cultures, the concentrations of intracellular ATP were approximately 43% and 27.7% less than that of the control, respectively. The specific activity of phosphofructokinase, one of the rate limiting enzymes of the glycolytic pathway, increased by a factors of 3.4 and 2.3, in comparison of the control respectively. With the specific activity of phosphofructokinase increased, the rate of glucose consumed increased by a factor of 3.6 and 2.4 compared with the control, and the rate of pyruvate produced increased by 17% and 8.5% respectively. The specific activities of hexokinase and pyruvate kinase were not affected by the addition of rotenone or antimycin A. Furthermore, the concentration of intracellular ATP decreased by 64.3% upon addition of 0.05 mg/L oligomycin to the cell culture, and the growth of Torulopsis glabrata was ceased when 0.4 mg/L oligomycin was added to the culture broth at 24 h. Both the rate of glucose consumed and the rate of pyruvate produced were enhanced with increasing the concentration of oligomycin ( ＜ 0.6 mg/L) in the cell cultures. As a result, the activity of phosphofructokinase (r2=0.9971), the rate of glucose consumed (r2=0.9967) and the rate of pyruvate produced (r2=0.965) were enhanced by a decrease in the energy level of the cell (the concentration of intracellular ATP). Increase of the rate of glucose consumed rooted in elevation of the specific activity of phosphofructokinase (r2=0.9958) and pyruvate kinase (r2=0.8706). These results are the first answer to the fundamental question of what controls the flux through glycolysis in Torulopsis glabrata.]]> LIU Li-Ming,CHEN Jian,LI Hua-Zhong and LI Yin 1308true HE Guo-Ping,ZHANG Si-Zhong,WANG Ying-Cheng,XIAO Cui-Ying,MA Yong-Xin,XU Wen-Ming,DING Lan,TAO Da-Chang,SUN Yan and CHEN Yu-Juan 1307true LSD1-like proteins are a family of plant-specific transcription factors that contain a specific class of C2C2 type zinc finger domain. Two members of this family have been identified and proved to control plant programmed cell death (PCD) in Arabidopsis. Here, a novel rice LSD1-like gene was cloned from a cDNA library and designated as OsLSD1 (for Oryza sativa LSD1). The cDNA is 988 bp in size, containing an open reading frame of 432 bp. The predicted OsLSD1 protein (143 amino acid residues) contains three internally conserved zinc finger domains. Southern blot indicated that OsLSD1 is a single-copy gene in rice. Furthermore, the OsLSD1 gene was expressed constitutively in rice root, stem and leaf. Using Bioinformatic tool, five and seven LSD1-like genes were identified from the databases of Arabidopsis and rice, respectively. The gene structure and domain organization of these LSD1-like genes were determined. Phylogenetic analysis of their coding sequences and predicted amino acid sequences suggested these genes can be classified into two classes, and a gene duplication event was identified. Although there is no specific class of LSD1-like gene in Arabidopsis or rice, some rice-specific zinc finger domains are found.]]> WANG Li-Juan,TIAN Ying-Chuan and HE Chao-Zu 1306true Staphylococcus aureus by PCR and ligated with the pGEM-T vector and confirmed by DNA sequencing. The expression vector pET-28a-SEC2 was constructed to express rSEC2 protein in E.coli. Puried rSEC2 protein was taken to bioactivity analysis. The cloned sec2 gene contains 717 nucleotides coding 239 amino acids which was accordance with that reported in GenBank. Cloned sec2 sequence was submitted to GenBank and the Accession number is AY450554. Soluble rSEC2 protein could be expressed effectively in BL21(DE3) and puried by Ni-NTA. Western blotting, PBMC proliferation assay and in vitro anti-tumor assay revealed that rSEC2 has the function of native SEC2. It was concluded that the whole sec2 gene was cloned and expressed in E.coli. Puried rSEC2 protein was proved to have the equivalent function of native SEC2.]]> XU Ming-Kai,ZHANG Cheng-Gang,ZHOU Ya-Feng and ZHANG Xian-En 1305true tert transcript in 60% (6/10) of matured oocytes (15 h-post hCG) had been not found, while the transcripts of housekeeping gene hprt located in all matured oocytes (10/10). No tert mRNA was present in single-copy embryo samples at early (20 h-post hCG, 6/6), late (30 h post-hCG, 8/8) pronuclear stage, or early 2-C stages (35 h-post hCG, 7/7), although the hprt mRNA transcriptions were found during this period. Interestingly, the embryos at early 2-C stage expressed of tert and hprt gene together in multi-copy (20 embryos/sample) embryo samples. Half of single-copy embryos (4/8) shown expression of two genes simultaneous at late 2-C stage (50 h-post hCG), while the other half of them (4/8) only transcript either tert (2/4) or hprt gene (2/4). Expression of tert and hprt genes were detected simultaneous in all single-copy embryos developed at 4-C (65 h-post hCG, 4/4), 8-C (75 h-post hCG, 4/4), morulae (93 h-post hCG, 4/4) and blastocyst (118 h-post hCG, 4/4) stages and the level of transcription increased. These results suggested that tert gene was began to express in embryos at early 2-C stage during the process of embryonic genome activation and the level of transcription was increased during embryogenesis.]]> WANG Yu,YAO Hui,AN Xiao-Rong,CHEN Yong-Fu and GOU Ke-Mian 1304true ZHAO Wei-Dong and CHEN Yu-Hua 1303true LAI Xiao-Qin and HUANG Li 1302true JIANG Xiu-Yun,WANG Chun-Feng,WANG Chun-Fang and HE Zhao-Yang 1301true SUN Jing,CHEN Hong,YANG Ying,XIA Hong-Fei and PENG Jing-Pian 1300true ZHOU Jia-Liang,HUANG Chao-Qun,WU Shi-Liang,SUN Qi-Chang,LEI Wei,JIN Mei-Fang,JIANG Zhi and SHEN Hong-Jie 1299true LI Shang-Wei,WEN Jian-Jun,LIU Shi-Gui and LONG Zhang-Fu 1298true 2 is a member of mammalian secretory phospholipase which belongs to phospholipase A2 superfamily. It has been expressed in a form of inclusion bodies in E.coli. It could not refold efficiently as human pancreatic phospholipase A2 (group Ⅰ B) merely by diluting the protein unfolded in 8 mol/L of urea. The refolding reaction of human group Ⅹ phospholipase A2 in vitro was depended on temperature, pH and protein concentration. A number of additives have been tested, among which L-arginine was the most efficient effector in improving the refolding of human group Ⅹ phospholipase A2, and its structural analogs, L-citrulline, had less effect. L-lysine and L-arginine methyl ester could not improve phospholipase A2 refolding. In addition, L-arginine inhibited the formation of aggregates and of intramolecular disulfide bonds. These results showed that both carboxyl and guanidyl residues of L-arginine were essential for improving protein refolding.]]> CHENG Hong-Qiang,ZHANG Chen,HU Hong-Yu and XU Gen-Jun 1297true WANG Xue-Feng,WANG Yi,JIANG Yan and MA Hui 1296true + channel. In the viewpoint of statistic thermodynamics, the two most readily permeant ions, K+ and Rb+ ions, can pass through the selectivity filter. In contrast, Na+ ions, which prefer staying in site 2, can not pass through the channel. The energy barrier difference between the K+ and Na+ is 27.4 kJ/mol which induced the permeant ratio of 15 000. The energy barrier difference of Rb+ ions is higher than that of K+ ions which is corresponding to the lower permeant of Rb+ ions. The results are qualitatively consistent with the experimental data. It is remarkable that the system, involved in 269 atoms, is much lower than the MD simulation which is about 41 000 atoms.]]> AN Hai-Long,ZHAN Yong,ZHANG Su-Hua,ZHAO Tong-Jun and LIU Jin-Wei 1295true NING Hong-Xiu,RONG Yu,ZHANG Yuan-Jiang,REN Fang-Li and CHANG Zhi-Jie 1294true DENG Hong-Wei,GUO Yan,SUN Ye and XU Yu-Hong 1293true CHEN Zuo-Zhou,XUE Cheng-Hai,ZHU Sheng,ZHOU Feng-Feng,XUEFENG BRUCE LING,LIU Guo-Ping and CHEN Liang-Biao 1292true XU Ping-Yong,BAI Li,TIAN Wei and XU Tao 1291true ZHOU Ji-Yong,Ye Ju-Xiu,Ye Wei-Cheng,Chen Qing-Xin,Zheng Xiao-Juan and Guo Jun-Qing 1290true ZHANG Kun-Lin,ZHANG Jing and LUO Jing-Chu 1289true SHEN Yan-Qin,LI Kang-Sheng,E. MOZE and P. J. NEVEU 1288true LI Xia- Chun,DUAN Ping,WANG Ze-Fen,ZHANG Jun-Xia,ZHANG Qi and WANG Jian-Zhi 1287true 1 α -subunit was obtained from Elumus sibiricus L. cv.'chuancao No.2' by reverse transcription-polymerase chain reactiom (RT-PCR) differential display. Full-length cDNA of 1 754 bp was cloned by 5 ′ RACE. The atpA has an open reading frame (ORF) of 1 518 bp that encodes a precursor protein of 505 amino acid residues. The deduced amino acid sequence exhibits 95%, 94% and 94% positional identity with atpA of wheat, rice and corn, respectively. Northern hybridization analysis on mRNA from treatment at 2 ℃ and post-treatment, in total of 13 time stages, showed that its RNA transcript was strongly inhibited after 12 h of chilling stress, whereas it was clearly higher than control during 4~8 h of chilling stress and 16~24 h after removing stress. This result provides new clue for revealing the CF1α -subunit role to ATPase in plant defence against chilling stress]]> HE Wen-Xing,XU Ying,TANG Lin,WEI Qin,LI Jing and CHEN Fang 1286true SHEN Yi-Jun,PAN Wei,XU Yan,WANG Zheng,YANG Tong,TAN Jing-Wei,WU Jin-Song,WU Fang,CAO Feng and LIU Yan-Jun 1285true ZHOU Yan-Hong,ZHANG Bi-Cheng,ZENG Zhao-Yang,ZHANG Wen-Ling,FAN Song-Qing,LI Xiao-Ling,XIONG Wei,CAO Li,LI Wei-Fang and LI Gui-Yuan 1284true WU Lei,HUANG Ming-Hui,ZHAO Jian-Long and YANG Meng-Su 1283true CUI Jie-Feng,LIU Yin-Kun,DAI Zhi,ZHOU Hai-Jun,SONG Hai-Yan,ZHANG Li-Jun,ZHANG Yu,SUN Rui-Xia,CHEN Jie and TANG Zhao-You 1282true YIN Dong-Mei,SHI Jian-Hua,GU Jian-Lan,SHEN Qin,GONG Cheng-Xin and LIU Fei 1281true YE Xun,LU Qin,ZHAO Yi,REN Zhen,MENG Xia,GE Sheng-Fang,QIU Qi-Hong,TONG Yong,LIEBER ANDRE,LIANG Min,HU Fang and CHEN Hong-Zhuan 1280true NI Chun-Sheng,SUN Bao-Cun,FENG Yu-Mei,ZHANG Dan-Fang,LI Xiao-Qing,DONG Liu-Huan and ZHANG Liang 1279true LI Hui,CAO En-Hua,HAN Bao-Shan and JIN Gang 1278true BI Li-Jun,ZHANG Xian-En,ZHOU Ya-Feng and ZHANG Zhi-Ping 1277true Lü Jun and LUO Liao-Fu 1276true YANG Jing,SONG Cun-Xian and LANG Xiao-Ou 1275true GAO Yan-E,ZHANG Ju,FAN Jiang-Bo,CHEN Zhong-Can and YAN Xiao-Jun 1274true XIA Yu-Feng,ZHAO Qian,YU Jing-Juan and AO Guang-Ming 1273true LI Dong,LI Jian-Qi,OUYANG Shu-Guang,WU Song-Feng,WANG Jian,Xu Xiao-Jie,ZHU Yun-Ping and HE Fu-Chu 1272true HUA Rong-Hong,TONG Guang-Zhi,WANG Yun-Feng and ZHOU Yan-Jun 1271true YUAN Chun-Xiu,FENG Xin-Gan,LIN Jiao-Jiao,LI Hao,LU Ke,SHI Yao-Jun,FU Zhi-Qiang,LIU Jin-Ming and CAI You-Min 1270true ZHANG Li-Neng,ZHA Xi-Liang and YU Qiang 1269true ZHANG Juan,QIU Qiang,TANG Jie,XIAO Zhong-Ju and SHEN Jun-Xian 1268true SHEN Li,CHEN Shao-Ping,QIN Yong-Wen and CAI Zai-Long 1267true PANG Li-Yan and RU Bing-Gen 1266true WU Zheng-Xing,XIE Li and XU Tao 1265true 1013 different sequences) . The selection scheme was constructed, and the experimental conditions, including library synthesis, formation of RNA-peptide fusion and RNA immobilization were optimized. Eight cycles have been processed and the results confirmed that the selected peptides could bind with thymidylate synthase mRNA specifically. Compared the amino acid sequences of the selected peptides with those from the initial random library, the basic and aromatic residues in selected peptides were enriched significantly, suggesting these peptide regions may be important in the peptide-TS mRNA interaction. As a novel in vitro selection approach, mRNA display technique would be developed as a powerful tool for isolation of functional peptides and proteins that could interact with immobilized targets with high affinity and specificity.]]> YAN Song,NIU Rong-Li,ZHANG Pei-Jun and LIN Xiu-Kun 1264true ZENG Zhao-Yang,XIONG Wei,ZHOU Yan-Hong,LI Xiao-Ling,LUO Xiao-Min,TANG Ke,LI Wei-Fang,Qian Jun and LI Gui-Yuan 1263true WANG Hai-Hua,XIE Ke,WU Kun-Lu and GUO Ze-Jian 1262true GONG Cheng-Liang,XUE Ren-Yu,CAO Guang-Li,SHI Xiao-Yan and SHEN Wei-De 1261true WANG Xiu-He and ZHANG Jing 1260true YIN Dong-Zhi,CAI Ji-Ye,ZHENG Qi-Chang,ZHAO Jing-Xian,LIU Mei-Li,DAI Yun,XIE Qiu-Ling and HONG An 1259true ZHOU Si-Bo,ZHANG Zhi-Hong,LIU Qian,ZENG Shao-Qun and LUO Qing-Ming 1258true ZHANG De-Xin,WU Jing,YANG Jing-Hua,REUVEN RABINOVICI and FAN Dai-Ming 1257true GAO Bo and JIN Gang 1256true LI Bo,FENG De-Yun,CHENG Rui-Xue,SUN Shu-Yan,HE Qiong-Qiong,HU Zhong-Liang,ZHENG Hui and WEN Ji-Fang 1255true 2006/9/15 0:00:00 LI Xian,XING Da,CHEN Xiao-Jia,TANG Yong-Hong,SUN Fen-Yong,WANG Ju,HONG An and WU Yun-Xia 1254true in vivo, adipoenctin gene knock-out and LacZ gene knock-in mouse model was constructed. Gene targeting strategy was designed to replace part of exon 2 and exon 3 of adiponectin gene with full length LacZ gene in frame with remaining upstream ATG and signal peptide sequence of exon 2. The targeting vector (Adipo-LacZ-XpPNT) was constructed and verified by restriction enzyme digestion and sequencing. CJ7 ES cells were transfected with targeting vector linearized by NotⅠ digestion, selected in the medium containing both G418 and ganciclovoir. Resistant clones were screened by PCR and further confirmed by Southern blot for correct homologous recombinants. Chimera mice were obtained by routing microinjection of homologous recombined ES cells into blastocysts. After mating, mice heterozygous and further homozygous for adiponectin knockout and LacZ gene knock-in were established. Expression of both endogenous adiponectin and exogenous LacZ gene in mouse tissues and sera were detected by RT-PCR, Northern-blot, Western blot and ELISA. The results show that adiponectin was disrupted at both mRNA and protein levels. LacZ gene is expressed exclusively in adipose tissue of mutant mice. Its expression profile is identical to endogenous adiponection. Unexpectedly, LacZ activity could not be detected in both adipose tissue and serum although LacZ protein can be detected in adipose tissue but not in serum of mutant mice. In conclusion, mice homozygous for adiponectin knockout and LacZ gene knock-in have been successfully constructed. Mutant mice display LacZ expression profile identical to endogenous adiponectin albeit neither LacZ activity nor protein can be detected in serum of mutant mice.]]> 2006/9/15 0:00:00 REN Wei- Hua,LI Xi-Hua,WANG Fang,QIAO Jian-Ou,DANG Su-Ying,KONG Hui,WANG Long,LU Shun-Yuan,SUN Xia,XU Guo-Jiang,FU Ji-Liang,FEI Jian and WANG Zhu-Gang 1253true Haematococcus pluvialis accumulates a highly valuable ketocarotenoid, i.e. astaxanthin up to 4% dry weight under stress conditions. Phytoene synthase is considered to be the first rate limiting enzyme in carotenoid biosynthesis pathway in H. pluvialis. The cDNA and genomic genes of phytoene synthase, i.e. psy from H.pluvialis were cloned and characterized. Result showed that psy had one open reading frame of 1 200 bp encoding a putative polypeptide of 400 amino acids which was interrupted by four introns. Phylogenetic analysis revealed that psy from green algae formed a monophyletic clade, and its closer relationship was higher plants. By using genomic walking approach, an approximate 1 kb 5′ flanking region of psy gene was cloned and a number of putative cis-regulatory elements were revealed. Fusing a 297 bp internal sequence (－297 to －1 bp from the translation initiation codon of psy) with the reporter gene, i.e. lacZ before attempted introducing the construct into the green alga via particle bombardment resulted in lacZ transient expression.]]> 2006/9/15 0:00:00 LIANG Cheng-Wei,ZHAO Fang-Qing,QIN Song,TAN Cong-Ping,WEI Wei and MENG Chun-Xiao 1252true 2 AR) is one member of G protein coupled receptors (GPCRs), which is a key pharmaceutical target in the treatment of asthma. Evolutionary trace (ET) method was employed to analyze AR sequences and 44 conserved residues were identified. Then molecular dynamics (MD) simulation of the β2 wild-type receptor, D130N active mutant and D79N inactive mutant were carried out and tried to explore the structural/dynamic features characterizing functionally different states of the receptor, by means of investigating ET identified conserved basic residues in the wild-type receptor and its two mutants. Particularly, it was found that the departing of D130 from R131 of DRY motif and approaching to K149 are highly correlated with the receptor activation, and the movement of helix 2, 4 and 6 upon receptor activation is inferred from the observation that R151 and K270 interact with other residues in the receptor active state on the basis of little change of the side chain orientations. The results might provide further insights into the activating mechanism of β2 AR mutants, as well as the molecular bases of the diseases induced by the mutations of the receptor.]]> 2006/9/15 0:00:00 CHEN Xiang,LIU Shu-Qun and SUN Zhi-Rong 1251true 2006/9/15 0:00:00 SHI Dong-Yun,LIU Shan-Lin,LI Hao-Ran,SHEN Xin-Nan,YU Pei-Zhong,CHENG Jian and GONG Xing-Guo 1250true 2006/9/15 0:00:00 WANG Ling,LU Cun-Fu and ZHANG Xu-Jia 1249true Sulfolobus solfataricus and Sulfolobus shibatae, especially in the six conserved motif sequences, which are known to be closely related to the key function of ligase. Recombinant JP2 ligase showed high activity in nick-joining reaction. It was the most active when Mn2+ present as divalent metal cofactor rather than Mg2+ and Ca2+. etc.. Assay of thermostability over a range of temperatures showed that at 50～80℃ the enzyme displayed relative high activity. Further thermostability experiment indicated that the activity of JP2 ligase could last for a long time at 80℃ and 85℃, however, at 90℃ and 95℃, it became unstable quickly. An investigation on the acquired thermotolerance of recombinant JP2 ligase was done by applying a chaperonin known as TF55 in thermophile on JP2 ligase reaction. Result showed that TF55 could not help in improving thermostability of ligase at 85℃. The possible reason might be that at 85℃ in vitro, the chaperonin itself was denatured.]]> 2006/9/15 0:00:00 LAN Hai-Yan,LIU Chun and HENDRY PHIL 1248true Sry in sex determination, the siRNA technique was used to silence the Sry expression in mouse embryos. Two siRNA expression vectors were synthesized, pSilencer4.1/Sry217 and pSilencer 4.1/Sry565. The siRNA expression vectors were injected into gestated mouse through tail vein. The sex was identified by PCR, and the expression level of Sry gene was determined using RT-PCR technique in the male mouse embryos at 11.5 dpc(days post coitum). The time- and dose -dependent effects was studied. The results confirmed that one of the siRNA expression plasmids, pSilencer4.1/Sry565, could inhibit the Sry gene expression significantly. Eighty-five percent inhibition was obtained when injecting 20 μg siRNA vector at 9.5 dpc.]]> 2006/9/15 0:00:00 ZHAO Jin-Hong,DU Wei-Hua,DAO Er-Ji,PEI Jie,ZHANG Li and LIN Xiu-Kun 1247true 2006/9/15 0:00:00 XU Shang-Long,LI Di-Chen,XIE You-Zhuan,LU Jian-Xi,LU Bing-Heng,ZHU Zhen-An,TANG Ting-Ting,DAI Ke-Rong and WANG Zhen 1246true ATP antagonist glibenclamide (glyb). By applying the glyb, the initial slight constriction (ISC) is obviously reduced, most of the ISC (10 μmol/L: 74.5%; 100 μmol/L: 96.2%) even completely restrained. And the peak of large dilation (LD) in response to the pial arteries was enhanced to (53.8±19.3) % and (59.8±19.6) % of the control with 10 μmol/L and 100 μmol/L glyb, respectively. It can be suggested that KATP in nerve cells of pre and post-synaptically plays a lead role in inhibiting CSD-associated hyperemia.]]> 2006/9/15 0:00:00 YANG Yuan-Yuan,LI Peng-Cheng,ZENG Shao-Qun and LUO Qing-Ming 1245true D. melanogaster and C. elegans. But as an evoulutional conserved phosphatase, little was known on its function in mammalian cells. siRNA expression vector was used to suppress PP4 expression in A549 and clonal cell lines was obtained in which the expression of PP4 was inhibited. The morphology, growth and mitosis process of the cells were investigated. It was found that cell growth and proliferation were greatly inhibited and many cells became multinuclei comparing with the control cells. Further study showed that the mitosis and cytokinesis became abnormal in PP4 silencing cells. It was proposed that PP4 played an important role in mitosis and cytokinesis. PP4 expression suppression resulted in multinucleus, then inhibited the proliferation of cells.]]> 2006/8/14 0:00:00 HUANG Xiu-Qing,LONG Zhi-Tao,NING Li-Feng,SUN,Ling-Ling and Sang Jian-Li 1244true E.coli and purification methods have been used to obtain the fragments of GDNF, GDNF(ΔN39), GDNF(ΔN39)-R9. Using primary cultured dopaminergic neurons and PC12 cells with transfected with GFRα1 and Ret to observe their biological function and cytotoxicity. Using B-Endo3 cells and Transwell method to analyze their delivery across the cellular membrane and blood brain barrier. The results show that GDNF(ΔN39)-R9 has the same neurotrophic function with wild GDNF and nearly no cytotoxicity to dopaminergic neurons and PC12-GFRα1-Ret cells and can get through effectually the cellular membrane and simulacrum of blood brain barrier with matrigel and B-Endo3.]]> 2006/8/14 0:00:00 Cui Min,Pan Xin and Wan Li-Mei 1243true in vitro/in vivo were examined successfully with GST pulldown and coimmunoprecipitations.It was shown that MT1E would be a new regulatory protein of XBP1. These screened proteins by yeast two-hybird system were closely correlated with liver fundamental metabolism,protein synthesis and transport,cell proliferation and apoptosis. The results mentioned above contributed to reveal the XBP1 biological function, and brought some new clues for further exploration of the expressing and regulating mechanism of XBP1.]]> 2006/8/14 0:00:00 Guo Feng-Jin,SONG Fang-Zhou,YI Fa-Ping and Cheng Hai-En 1242true 10-dB value and symmetry index > 0 of FTC, the probability of CF shift toward HFS of FTC is higher than other neurons. The results suggest that the experience can alter CF in rat auditory cortical neurons. These findings also provide evidence for further study of plasticity mechanisms in sensory function of the brain.]]> 2006/8/14 0:00:00 Wang Fang,YANG Wen-Wei,TAN Jiang-Xiu,PENG Yin-Ting,ZHANG Ji-Ping and SUN Xin-De 1241true LAG1 was identified as a gene that is differentially expressed during the yeast replicative life span and was shown to play a role in determining yeast longevity. The cDNA of rat LASS1, the mammalian homolog of yeast LAG1, was cloned from rat cerebral cortex and sequenced, which is different to the predicted sequence in the GenBank. Sequence analysis revealed that this cDNA clone contains an open reading frame of 1 053 bp. The deduced amino acid sequence has 350 residues and shares a predicted Lag1p motif and a TLC domain conserved in Lag1 proteins. Total RNAs were isolated from rat cerebral cortices at varying ages: newborn, one month, six months, twelve months, and twenty-four months. Semi-quantitative RT-PCR and Northern blot analysis were performed to analyze the LASS1 expression level in rat cerebral cortex tissues at varying ages. Senescence-associated β-galactosidase (SA-β-gal) activity was firstly used as a biomarker for assessing senescence in rat neurons. The results showed that LASS1 expression was upregulated from newborn to adult rats (1～6 month) and declined in aged cortex. SA-β-gal staining positive neurons significantly increased in the aged cerebral cortex. The age-related expression alternation of LASS1 in rat cerebral cortex provides an important clue in exploring the role of LASS1 in mammalian neuron aging.]]> 2006/8/14 0:00:00 Wang Bao-Heng,Fu Yu Cai,SHI Gui-Zhi,XU Ming-Yan,GENG Yi-Qun,XU Xiao-Hu and XU Jin-Jie 1240true r = 0.9997). The cytotoxic activity of VCR encapsulated PEG-PE micelles demonstrated high inhibitory effect on MCF-7 cell growth compared to the free VCR. It is concluded that VCR encapsulated in PEG-PE micelles were stable and have high drug entrapment efficiency and high effect of growth inhibition in vitro. PEG-PE micelles may be promising vectors for delivery of VCR into tumor in vivo.]]> 2006/8/14 0:00:00 Lin Hong-Ying,LU Xiao-Yan,TANG Ning and LIANG Wei 1239true 2006/8/14 0:00:00 Li Ai-Ying,Wen Jin-kun and Han Mei 1238true E.coli strain BL21(DE3) with an improved expression vector pASK75-EX as inclusion body. The denatured inclusion body was purified with Ni-NTA chelate agarose, and then re-natured by dialysis. FACS and MTT assays indicated that the re-natured fusion immunotoxin B-L-SEC2 could target the HER-2 over-expressing breast tumor cell SK-Br-3 in vitro, and inhibit the growth of SK-Br-3.]]> 2006/8/14 0:00:00 XU Ming-Kai,ZHANG Cheng-Gang,ZHANG Hui-Wen,ZHOU Ya-Feng,ZHANG Xian-En and LIU Li 1237true 2CO3 groups. The level of hyperphosphorylated tau and the expression of 2PP-2A was measured respectively by Western blot. It is suggested that GSK-3 activity increases, PP-2A activity and expression decrease, and hyperphosphorylated tau be produced at Ser198/Ser199/Ser202 and Ser396/Ser404 in DM rats cortex. After the DM rat were treated with Li2CO3, the inhibition of GSK-3 activity and the improvement of PP-2A activity were found, and hyperphosphorylation of tau at Ser198/Ser199/Ser202 and Ser396/Ser404 were deduced. These studies firstly suggested that an increase of GSK-3 activity might inhibit PP-2A activity, and which produce hyperphosphorylated of tau at Ser198/Ser199/Ser202 and Ser396/Ser404 in DM rat cortex in common.]]> 2006/8/14 0:00:00 QU Zhong-Sen,ZHAO Yong-Bo,LIU Wen-Wen and WANG Jian-Zhi 1236true 2006/7/17 0:00:00 LI Bo,WANG Da-Peng,FANG Wen-Gang and CHEN Yu-Hua 1235true 2006/7/17 0:00:00 SHI Jun,LI Xu,LU Fen,CHEN Xiao-Qian,WANG Jian-Zhi and GONG Cheng-Xin 1234true Oryza sativa L. Zhenhui 249Y) and its wild type (Oryza sativa L. Zhenhui 249W). The PSⅠ-LHCⅠ super-complexes, LHCⅠ-less PSⅠ, ATP synthase, Cytochrome b6f, CP43-less PSⅡ, trimeric and monomeric LHCⅡ were resolved. 24 proteins, which belong to the subunits of four major photosynthetic apparatus were identified. Comparative study indicated that the mutant had moderately decreased amount of LHCⅡ, greatly decreased amount of LHCⅠ, and increased amount of PSⅡ reaction center and ATP synthase. Western blotting verified BN/SDS-PAGE analysis. These results provided clues about molecular foundation of higher photochemical efficiency and light stability of the mutant, and showed that the improved BN/SDS-PAGE is suitable for not only membrane protein complex separation, but also for comparative study of complex subunits in different physiological condition or between mutant and its wide type.]]> 2006/7/17 0:00:00 CHEN Xi,CUI Xiang-Ju,YING-XIN ZHAO and ZHANG Wei 1233true 2006/7/17 0:00:00 LU Ming,SHI Bao-Chen,LI Xue-Ling,LU Jun-Hong,CHEN Run-Sheng and HU Jun 1232true Schistosoma japonicum (SjHGPRT) were amplified by the anchored PCR with 2 pairs of primer that were designed according to the published incomplete SjHGPRT EST and the sequence of multiclone sites of library λgt11 vector. Sequence analysis indicated that this fragment, with an identity of 82% to hypoxanthine-guanine phosphoribosyltransferase of Schistosoma mansoni (SmHGPRT), contained a complete open reading frame(ORF). The deduced amino acid sequence showed 83% identity to that of SmHGPRT. This fragment was cloned into the prokaryotic expression vector pQE30, and subsequently sequenced and expressed in Escherichia coli. SDS-PAGE revealed that M of the recombinant protein was about 28 ku. Western-blot analysis showed that the recombinant protein was recognized by the polyclonal antisera from rabbits immunized with Schistosoma japonicum adult worm antigen. Mice vaccinated with recombinant protein revealed significant worm burden, liver eggs per gram (LEPG), fecal eggs per gram (FEPG) and intrauterine eggs of the female worms reduction percentage, compared with the controls. Taken together, the SjHGPRT full-length cDNA can be cloned and expressed in E.coli as a recombinant protein that elicited immunity against the challenge infection with Schistosoma japonicum, indicating its potential as a partial protection vaccine candidate.]]> 2006/7/17 0:00:00 YU Jun-Long,WANG Shi-Ping,Hezhuo,DAI Gan,Liwenkai,JIANG Xiao-Xin,ZENG Shao-Hua,XIAO Xiao-Qin,XU Shao-Rui,Lv Zhi-Yue,PENG Xian-Chu,ZHOU Song-Hua and LIU Xue-Qin 1231true Streptococcus mutans UA159 was cloned and expressed in E. coli, and purified to homogeneity. The FPLC size exclusion chromatography analysis reveals that the S. mutans dCMP deaminase forms hexamer in solution. The protein was crystallized using hanging drop vapour-diffusion method and diffracted to a resolution of 3.1Å. The diffraction data were collected at BSRF beamline 3W1A. The crystals belong to P213 space group, with unit cell parameters a=b=c=113.2Å, α=β=γ=90°. Assuming there are two subunits per asymmetric unit, the Matthews coefficient is 3.6Å3·Da-1. This is the first crystallization report of the wild-type deoxycytidylate deaminase.]]> 2006/7/17 0:00:00 HOU Hai-Feng,GAO Zeng-Qiang,XU Jian-Hua,XU Rui,LI Li-Qin,LI Lan-Fen,LIANG Yu-He,SU Xiao-Dong,LIU Peng,XIAN Ding-Chang and Dong Yu Hui 1230true 2006/7/17 0:00:00 YU Li-Ping,WANG Xiao-Yan,LI Xiang-Yao,ZHANG Ji-Ping and SUN Xin-De 1229true 1 transgenic individuals were detected by polymerase chain reaction and Southern blot hybridization, which showed that the transgenic rate of mouse F1 offspring was 41%. The transferred gene was integrated into the host genome and could be transmitted to its offspring. When the positive F1 individuals were mated with the wild type ICR mice, the F2 individuals had a transgenic rate of 37%. The results indicate that the high efficiency of gene transfer and the limited number of manipulations make the method suitable for creating a large number of transgenic animals, especially, for producing domestic animals.]]> 2006/7/17 0:00:00 HE Xin,QI Bing,LIU Gui-Sheng,YU Wei-Dong and CHEN Qing-Xuan 1228true 58) with fibroblast growth promoting activity in silicotic rat, which had high sequence homology with members of the mammalian chitinase protein family has been previously purified and characterized. Bioinformatics analysis showed that several human EST clones from pooled colon, kidney or stomach matched the rat protein sequence. Thereafter clone from human kidney RNA samples with several pairs of primers was managed and a set of sequences was obtained, whose cDNA and amino acid sequences have high similarity with each other and several human chitinases in GenBank. Comparison with human genome sequence suggests that these molecules may be from variant transcripts of same a Pre-mRNA. Here the characterization of these sequences is reported.]]> 2006/7/17 0:00:00 CHEN Xiao-Hua and CAI Guo-Ping 1227true VvSUC27) of grape sucrose transporter was inserted into an expression vector pBI121 in the anti-sense orientation under the control of the cauliflower mosaic virus 35 S promoter, and subsequently transferred into tobacco(Nicotiana tobacum cv. Samsun). The plantlets of the transformed anti-sense VvSUC27 cDNA tobacco were grown normally on MS media containing 20g/L sucrose, but it was found that roots of the transformants developed slowly and the leaves have more chloroplasts when observed under slice. Sucrose content in the roots of the transformants was only 51% of the controls. The capability of sucrose uptaking and transporting into leaves among the transformed tobaccos was greatly decreased.]]> 2006/7/17 0:00:00 ZHANG Ya-Li,MENG Qing-Yong,QU Gui-Qin,LIN Xi-Jin,LU Jiang and LUO Yun-Bo 1226true 2006/6/14 0:00:00 Liu Hua-Ying,Luo Xiao-Min,Niu Zhao-Xia,Peng Cong,Li Gui-Yuan,Zhou Ming,Zhou Yan-Hong,Li Xiao-Ling,Li Wei-Song and Xiang Bo 1225true 2006/6/14 0:00:00 YANG Ming,WANG Chun-You,LIU Tao,WANG Yang,LIU Wei and LI Ming-Tao 1224true 2006/6/14 0:00:00 Zeng Zhu and Sun Dagong 1223true 2006/6/14 0:00:00 Qiu Lang-Bo,Wang Guang-Yun and Wang Zheng-Zhi 1222true 2006/6/14 0:00:00 GUO Lin-Lang,GUO Ying,LAU Derick,XIAO Sha,XU Yin-Chao and SHEN Hong 1221true Pseudomonas aeruginosa apoazurin is an arguing problem. Recent published results indicated that this problem could be resolved by hypothesizing two native conformations coexisting in solution. Urea-induced unfolding of apoazurin was investigated further using intrinsinc fluorescence and CD spectra. Equilibrium unfolding curves in urea could be depicted with an apparent two-state transition, but a biphase kinetic process. The fast unfolding process is finished within a few seconds as monitored by stopped-flow fluorescence intensity, whereas the slow process requires several hours for unfolding at high concentration of urea at room temperature. The mu values for the fast and slow phases are 2.24 and 2.45 kJ·mol－1·M－1 respectively. The difference between their unfolding activation energy is 22 kJ·mol－1. The time-resolved fluorescence emission spectra as well as the circular dichroism spectra just after manual mixing of protein and denaturant could be simulated by superposition of the spectra of the native and fully unfolded protein with the same coefficient of 0.37. This strongly suggests the three-state mechanism with a partially unfolded intermediate on its pathway is inadequate. The hypothesis of two native conformations coexistence is a reasonable selection.]]> 2006/6/14 0:00:00 Zhong Jin and Zhang Hongjie 1220true E.coli system. The results of toxcity studies indicated that the acute toxicity in guinea pig and cytotoxicities of mutant E154D increased slightly in compared with those of recombination wild toxin, and contrarily, those of E154R decreased obviously.]]> 2006/6/14 0:00:00 GaoChuan,Song Yun-yang,Wang Hui-Fang,ZhangJing and Hu Yalan 1219true 2(＋)/CREG] and antisense-CREG [pLXSN(－)/CREG] retroviral vectors and positive clone was abtained by G418 selection. The cells infected with pLNCX2(＋)/CREG take on the differentiated phenotype and overexpression of CREG can enhance SM α-actin expression accompanied with increase of the total RhoA and nuclear serum response factor (SRF) expression detected by immunofluorescence and Western blot. And treated with a selective Rho kinase inhibitor Y-27632, the expression of SM α-actin and nuclear SRF protein induced by CREG was effectively reduced. Meanwhile, the opposite results were observed in the CREG-depression HITASY cells. And immunoprecipitation assays indicate that as a secreted protein, CREG can be detected in the media of pLNCX2(＋)/CREG cells and can interact with insulin-like growth factorⅡ/mannose 6-phosphate receptor (M6P/IGF2R). Furthermore, that using okadaic acid (an inhibitor of the protein phosphatases 2A) to decrease the number of M6P/IGF2R at the cell surface can significantly inhibit the expression of total RhoA, nuclear SRF and SM α-actin induced by CREG-overexpression. Taken together, as a secreted protein, CREG can enhance the expression of SM α-actin by interaction with M6P/IGF2R and perform a pivotal role in the process of VSMCs differentiation and phenotype modulation.]]> 2006/5/12 0:00:00 HAN Ya-Ling,XU Hong-Mei,YAN Cheng-Hui,Hu Ye,KANG Jian and LIU Hai-Wei 1218true P value of hypergeometric cumulative distribution of modules and the disturbance analysis on the data, including adding, removing and rewiring interactions, are employed to evaluate the prediction quality and robustness of the method. The results show that MCM has high prediction precise rate and coverage, and it is robust to high false-positive data and missing data. The predicted results of unknown proteins with high prediction precise rate can be instructive in biological analysis and the algorithm can be generalized to other networks with the similar structures.]]> 2006/5/12 0:00:00 LU Hong-Chao,SHI Qiu-Yan,SHI Bao-Chen,ZHANG Zhi-Hua,ZHAO Yi,TANG Su-Qin,XIONG Lei,WANG Qiang and CHEN Run-Sheng 1217true in vitro. To study the osteogenesis ability of hMSCs treated by shock wave in porous hydroxyapatite (HA) in vivo, primary hMSCs of SW group and control group were cultured in the porous HA for 2 weeks and then implanted into subcutaneous sites of nude mouse. These implants were harvested and prepared for the biochemical analysis of alkaline phosphatase activity by AKP kit, histological analysis of decalcified and undecalcified sections and morphology by scan electric microscope (SEM), as well as osteocalcin mRNA expression by RT- PCR 4 weeks and 8 weeks after implantation. It showed that cells of SW and control group almost covered the rough surface of HA before implantation and the extracelluar matrix of SW group was abundant by SEM photomicrograph . The histological analysis and SEM photomicrograph showed active bone formation 4 weeks and 8 weeks after implantation, as well as tetracycline labeling under fluoroscopy analysis in SW group. Alkaline phosphatase in supernatants of the implants detected 4 weeks and 8 weeks after implantation in SW group was higher than in control group (P < 0.001). The expression of osteocalcin mRNA was found 4 weeks and 8 weeks after implantation in SW group. The results suggest that postnatal hMSCs treated by SW could form bone tissue in vivo using tissue engineering technique, and this new method would be applied in the field of bone tissue engineering.]]> 2006/5/12 0:00:00 HU Jun,XING Da,ZHANG Ai-Bin and ZHOU Jiang-Nan 1216true 2006/5/12 0:00:00 YANG Yan,HU Shu-Hong,ZHANG Jian-Hua,ZHANG Mu-Xun and GONG Cheng-Xin 1215true Tm of 64.6℃ and the thermal stability of TIM was decreased in the presence of low concentrations of guanidine. The possible unfolding pathway of the dimeric TIM was discussed. It shows that the secondary and tertiary structural changes of TIM occur simultaneously during guanidine denaturation and the unfolding of dimeric TIM follows an apparent two-state transition without detectable dissociation model.]]> 2006/5/12 0:00:00 Liu Jiang-Hong and Zhou Jun-Mei 1214true Helicobacter pylori (Hp) urease (Ure) beta subunit were chosen as target protein. The advantageous linear epitope of these proteins was analyzed and screened. The multiple antigenic peptides(MAPs) containing the selected sequence were synthesized by Fmoc solid phase method,purified through high performance liquid chromatography(HPLC) and printed on nitrocellulose membrane in microarrays by computer-controlled high-speed robotics. The nitrocellulose membrane was blocked with 2% bovine serum albumin solution. The MAPs microarray was finished by assembling the membrane with plastic outer shell. Some microarrays were selected at random for quality control identified by control sera and compared with ELISA method. 4 MAPs were screened out, synthesized and identified. The result of positive and negative sera of Hp and HCMV detected by MAPs microarray were consistent with the control sera. In the clinical trial of 120 random sera, the microarray performed almost equally with ELISA method using recombinant antigen and microbial lysate antigen. The sensitivity and specificity of Ure-1, Ure-2 and PP150 MAPs were higher than 90%. The CV were lower than 7% among microarrays showing a good repeatability. It can be concluded that a MAPs microarray analytical platform which has a vast application prospect in assistant design of peptide vaccine was primarily established.]]> 2006/5/12 0:00:00 XU Xiao-Jie,ZHANG Hu-Ming,LI Ding,HAN Feng-Chan,ZHANG Wen-Hong,GUO Hui-Fang,WEN Dong-Qing,LUO Jin,YAN Xiao-Jun and KANG Lei 1213true α atom. The off-lattice model is adopted and the relative entropy is used as a minimization function to predict the tertiary structure of a protein. A new contact intensity function is given to consist with protein design research based on the relative entropy. Testing on several real proteins from Protein Data Bank (PDB) shows the good results obtained with the model and method. The root mean square deviations (RMSD) of the predicted structures relative to the native structures range from 0.30 to 0.70 nm. A foundation for studying protein design using the HNP model and the relative entropy was made.]]> 2006/5/12 0:00:00 SU Ji-Guo,WANG Bao-Han,JIAO Xiong,CHEN Wei-Zu and WANG Cun-Xin 1212true 2006/5/12 0:00:00 ZHAO Liang,LIU Li,WANG Shuang,LI Zu-Guo and DING Yan-Qing 1211true 2006/5/12 0:00:00 ZHAO Mei-Rong,CHEN Zhi-Qiang,QIU Wei,LI Yu-Xia and WANG Yan-Ling 1210true NTR (or TROY). Recently, LINGO-1 is identified as another essential component and regulator for the Nogo-66 receptor/p75 signaling complex. LINGO-1 is restricted to express in CNS, neuronal LINGO-1 is shown to be involved in the signal transduction from three myelin proteins, and Lingo-1 in oligodendrocyte negatively regulates the differentiation and myelination of oligodendrocyte. To investigate the potential activity of LINGO-1 in neuronal apoptosis, LINGO-1-Fc fusion protein including the extracellular LRR and IgC2 domain, was used as functional antagonist to study its protective effect on low-potassium induced apoptosis of cerebellar granule neurons (CGNs). In judgement of the apoptotic nuclei stained by Hoechst, LINGO-1-Fc pretreatment for 2 h significantly prevents apoptosis of CGNs. Although GST-LINGO-1 protein, including the LRR domain, binds to the CGN cultures in the same way with LINGO-1-Fc, it doesn't prevent the apoptosis of CGNs. These results indicate that LINGO-1-Fc fusion protein prevents low-potassium induced apoptosis of cerebellar granule neurons in certain conditions and this activity is probably IgC2 domain dependent.]]> ZHAO Xiang-Hui,JIN Wei-Lin,MI Sha and JU Gong 1209true HAN Ya-Ling,XU Kai,KANG Jian,YAN Cheng-Hui,TIAN Xiao-Xiang and LI Shao-Hua 1208true 6f complex and the ATP synthase complex are located in the thylakoid membrane. Here, the newly developed split-ubiquitin two hybrid system was used to investigate protein-protein interactions of thylakoid membrane proteins. The reaction center D1 protein of PSⅡ, which is encoded by plastid gene psbA, was used as the bait protein, while the D2 encoded by plastid gene psbD, another PSⅡ reaction center protein, and Cytb6 encoded by plastid gene petB, a component of Cytb6f complex, were used as prey proteins. The yeast two hybrid analysis showed that the D1 protein interacts with D2, but not with the Cytb6. This result confirms the protein interaction model of thylakoid membrane protein complex, and also means the availability of this system in detecting thylakoid membrane protein interactions. Thus, the split-ubiquitin two hybrid systems could provide an efficient tool to reveal the regulation mechanism of chloroplast proteins biogenesis.]]> ZHU Shu-Yong,CHI Wei and ZHANG Li-Xin 1207true vs (185.85±9.45) g, P < 0.05). Furthermore, the GH maintains on a stable level in group ppG-H6 and ppG-H4 compare to control by the end of experiment, which support that the injection of ppG-H6 and ppG-H4 plasmids caused the body-weight of rat increasing. The results imply that injection of ectopic expression of porcine growth hormone-releasing hormone (GHRH) plasmids is a possible way to enhancing animal growth.]]> MENG Qing-Yong,ZHANG Ya-Li,XIE Qi-Fa,GONG Xia and LI Ning 1206true CS-ACS2) was amplified from genomic DNA of 8 different sexual phenotypes of cucumber respectively (GenBank accession number is DQ115884～DQ115886 and DQ115875～DQ115879). 8 SNPs have been identified by sequences analysis between 3 monoecious lines and 5 subgynoecious lines and gynoecious lines, which including 4 A←→G and 4 T←→C transition. Of these 8 SNPs, one locus is in intron and 7 loci in exons. Of the 7 SNPs located in exons, 3 SNPs are non-coding SNPs and 4 SNPs are coding SNPs (cSNPs) of which 3 induced changes of encoding amino acid of ACC synthase. The results of SNPs from subgynoecious lines and gynoecious lines suggest that single nucleotide mutation events of CS-ACS2 might be correlated with the development of subgynoecious lines and gynoecious lines in cucumber. Furthermore, CAPS marker C-MT705 was developed for identifying elite subgynoecious cultivar MT-705, which could be valuable in cucumber breeding. Besides, the SNPs and CAPS markers obtained in the study enriched molecular markers of cucumber.]]> XIANG Tai-He,WANG Li-Lin,PANG Ji-Liang,HU Jiang-Qin,SHENTU Lian-Feng and WU Kai 1205true OsGME1 transcript was up-regulated in response to cold stress, and gibberellin might regulate L-ascorbic acid levels by affecting transcription of both OsGME genes.]]> DU Ying,YAO Yuan and LIU Jin-Yuan 1204true ZHAO Geng-Xiang,YUAN Cai,BIAN Chuan-Bing,JIANG Long-Guang,YE Xiao-Ming,HUANG Zi-Xiang and HUANG Ming-Dong 1203true in situ hybridization with the coding region of mUBAL showed that expression of mUBAL was presented in the early streak stage of mouse embryo and migrated anterior with anterior visceral endoderm (AVE) and then in the telencephalon. The gUBAL expression was also found in the brain of HH14，HH 16 and HH18 chick embryos. The conserved structures and expression patterns of UBAL suggested that UBAL may play important roles in the developing brain of embryo and adult tissues as E1 like enzyme.]]> XUE Peng,CI Hong-Liang,CHEN Wei and LI Yi-Ping 1202true Arabidopsis COI1 was isolated from rice (Oryza sativa L.) by RT-PCR and designated as OsCOI1. It encoded a protein of 595 amino acids. The similar F-box motif and 16 leucine-rich repeats were found in the deduced protein OsCOI1. OsCOI1 and COI1 showed high homology (74%) at amino acid level. Semi-quantitative RT-PCR and Northern blot analysis demonstrated that the expression of OsCOI1 in rice varied obviously after treatment with MeJA and ABA but was not affected by SA and ET, suggesting that the specific function of OsCOI1 in JA signal pathway and related ABA pathway.]]> HU Ting-Zhang,WANG Wei-Ping,CAO Kai-Ming and WANG Xi-Ping 1201true 12121 with unit-cell dimensions a = 76.39 Å, b=52.77 Å, c=27.12 Å. This dimeric structure was solved by molecular replacement and refined to R=0.109 for all reflections at a resolution of 1.4 Å. The extensively refined structure definitely shows that the cis peptide bond Pro9-His10 equally occurs in both molecule A and molecule B in the dimer. The observation manifested that this striking non-proline cis peptide is not related to crystal packing, but caused by certain intrinsic factors. The detailed analyses and comparison with the structure of BmK M8, which is homologous to M1 but has trans peptide bond 9-10, showed that the five-residue reverse (8～12) with a consensus sequence (-KPXNC-) may be the intrinsic structural element for the cis form of this peptide bond. A pair of well organized main-chain hydrogen bond between residues 10 in cis unit and 64 at C-terminus forms main tertiary interactions to stabilize this energetically unfavorable peptide bond.]]> HE Xiao-Lin,GUAN Rong-Jin,ZHANG Ying and WANG Da-Cheng 1200true WANG Chun-Fei,CHEN Ming,FUNG Man-Lung and GAO Tian-Ming 1199true HUANG Bo and LIU Jin-Yuan 1198true WANG Shi-Lei,HUANG Bo,ZHOU Yun and SUN Zhi-Rong 1197true LI Yan-Hua,LI Han-Jie,ZHAO Xing-Bo,LI Ning and WU Chang-Xin 1196true YI Hong,YANG YI-Xuan,CHEN Zhu-Chu,ZHANG Gui-Ying,ZHANG Peng-Fei,LI Jian-Ling,ZHU Guo and XIAO Zhi-Qiang 1195true 2006/3/16 0:00:00 Yuan Cai,BIAN Chuan-bing,HUAI Qing and HUANG Ming-Dong 1194true 11 level. The N1N2 region of gene Ⅲ was amplified and cloned into pTMI under the control of lac promoter to give pTMIN. Promoter trc was synthesized and replaced the lac promoter to give pTTMIN which permits the fusion expression of antigen with N1N2. To test its ability for fusion expression, gene code for ten-peptide of c-myc was synthesized and inserted into pTTMIN downstream to N1N2. After induction expression, the results of ELISA and SDS-PAGE showed that it has been expressed successfully. When pTTMIN was transfected into cell carrying antibody display vector p3MHHB3, it was copackaged into phage particles in 0.3% to 55% after rescuing with helper phage VCSM13.From the results it can concluded that the antigen expression vector was constructed successfully and it can be used for library-library screening in theory.]]> GAO Rong-Kai and WANG Yan 1193true WANG Dong,HUANG Shun-Jun,XING Li,GUO Xi-Qin,HAN Yue,LUO De-Yan,LI Min and WANG Xi-Liang 1192true CAO Yun-He,YUAN Xue-Feng,Wang Xiao-Xing,GUO Li-Hua,CAI Zhu-Nan,HAN Cheng-Gui,LI Da-Wei and YU Jia-Lin 1191true XIN Yan-Fei,ZHOU Xiang-Jun and WANG Yong-Xiang 1190true XU Li-Yan,LI En-Min,NIU Yong-Dong,CAI Wei-Jia,YUAN Hua-Min,CHANG Jing-Xia,SHEN Zhong-Ying and ZENG-Yi 1189true MA Yan-Xia,XU Bo,CUI Jing-Rong,WANG Li,WU Jun and LI Min 1188true SHI Jian-Yu,PAN Quan,ZHANG Shao-Wu and LIANG Yan 1187true DUAN Chang-Zhu,PU Shu-Ping,TSUTOMU MORI,HIDEO KOCHI and QIU Zong-Yin 1186true TIAN Ping-Fang,WANG Jian-Jun,WU Gang,LI Qun,LU Bao-Rong and HE Zu-Hua 1185true 0.05). －40 mV activated threshold potential and －30 mV peak potential for control group respectively dropped to －60 mV and －40 mV after irradiating 7 min. The half-activation voltage and the slope factor of the activation curves of Na+ channel were also changed by the laser's exposure. The former changed from (－42.091 ± 1.537) mV to (54.971 ± 1.846) mV (n＝8, P < 0.01) and the latter form (1.529 ± 0.667) mV to (2.634 ± 0.519) mV (n=8, P < 0.05). The results show that activation properties of Na+ channel are influenced by laser irradiating hippocampal neurons. Thus, depolarizing process of action potential is affected. And further physiological functions of neurons are altered as a result of low level laser irradiation.]]> QIAO Xiao-Yan,LI Gang and HE Bing-Jun 1184true ZHANG Xue-Feng,ZHENG Yi-Xin and CHANG Zeng-Yi 1183true LOU Li,GU Yu-Dong and SHOU Tian-De 1182true FENG Jing,SHEN Yi,YANG Dong-Ling and YAN Xi-Yun 1181true XIE Yi,ZENG Wei,JIA Wen-Xiang,YANG Fa-Long,YANG Wei-Qing,CHENG Xi,KANG Mei,WNAG Lan-Lan and ZHANG Zai-Rong 1180true DENG Min,HE Xiu-Sheng,LUO Qiao,ZHAO Shuai,ZENG Chao and LI Yan-Lan 1179true WANG Xiao-Yan,SHEN Shou-Rong,LIU Fen,LI Xiao-Ling and FAN Song-Qing 1178true LU Min,GONG Xing-Guo,WANG Chen-Hui,ZHENG Le,GUO Jian-Jun and ZHANG Shen-Feng 1177true ZHOU Hai-Jun,LIU Yin-Kun,CUI Jie-Feng,JIN Hong,WEI Li-Ming,FAN Hui-Zhi,GUO Kun and YANG Peng-Yuan 1176true 0.05). Then, positional cloning strategy, bioinformatics technology and reverse transcription-polymerase chain reaction (RT-PCR) were used to identify candidate TSGs and its cDNA fragments at 6q16.3～21, especially at the high frequency deletion region. Comparing with expression of normal peripheral blood mononuclear cell, EST screened in D6S1709-D6S301(GenBank Accession No.AA403058) was down-regulation in ten of fifteen childhood ALL (P < 0.05). Digital differential display showed that the expression levels of AMD1, PPIL6 and WASF1 were lower in cancer tissues than in normal tissues(P < 0.05). These findings may provide new clues in cloning of childhood ALL TSGs at 6q16.3～21.]]> KANG Rui,CAO Li-Zhi,YU-Yan,YANG Ming-Hua,ZHANG Zhao-Xia,GUO Bi-Yun,XIE Min,CHEN Ying,TAN Zhi-Hong,WANG Zhuo,HU Ting and Wu Xiu-Shan 1175true WEI Min,Tian Mei-Hong,CHEN Lin and Zeng Xian-Lu 1174true CUN Wei,ZHANG Ying,LIU Long-Ding,WANG Li-Chun,DONG Cheng-Hong and LI Qi-Han 1173true GUO Hui-Fang,ZHANG Wen-Hong,WEN Dong-Qing,HAN Feng-Chan,ZHANG Hu-Ming,LUO Jin and YAN Xiao-Jun 1172true 1～42 was stereotaxicly injected into rat hippocampus with reverse peptide Aβ42～1 as control. After 7 days post-injection, the expression of macrophage inflammatory protein-1α (MIP-1α) and its receptor (CCR5) in peripheral blood T cells was detected by real-time quantitative polymerase chain reaction (RT-qPCR). Brain sections were analyzed with immunofluorescence of CD3, VWF and CCR5. The results showed that Aβ1～42 deposits in rat brains led to significantly higher expression of MIP-1α in circulating T cells than Aβ42～1 did, while no increase of CCR5 expression was observed in circulating T cells of Aβ1～42-injected rats compared to Aβ42～1-injected rats. Furthermore, the expression of CCR5 was up-regulated by Aβ1～42 on rat brain microvascular endothelial cells (RBMEC). In addition, T cells were increased in abundance in Aβ1～42-injected brains compared with Aβ42～1-injected brains, scattered mainly in cortex and hippocampus. Treatment of Aβ1～42-injected rats with neutralizing antibody specific for MIP-1α dramatically blocked the enhanced T cells entry into rat brain. The results implied that the interaction between MIP-1α over-expressed in T cells and CCR5 on RBMECs may contribute to the Aβ1～42-induced circulating T cells migrating across blood-brain barrier.]]> 2006/12/15 0:00:00 GUO Da-Wen,MA Yi-Ran,FANG Wen-Gang and CHEN Yu-Hua 1171true 2006/12/15 0:00:00 ZHANG Hong-Mei,ZHOU Li-Jun,HU Neng-Wei,ZHANG Tong and LIU Xian-Guo 1170true in vitro though they had no effect on the acute insulin secretion. BTCe also effectively ameliorated the hyperglycemia of STZ-induced diabetic rats by a single plasmid injection into muscle of rats. It is supposed that BTCe promote the proliferation of PDX-1 positive cells or repair some signal transduction pathway. Perhaps the latter is more important.]]> 2006/12/15 0:00:00 LI Hong,ZHOU Wei-Bin,LI Feng-Ying,GU Yan-Yun,TIAN Jing-Yan,QIAN Lei,ZHANG Di,ZHOU Wen-Zhong,WU Guo-Ting,LUO Tian-Hong,LI Guo and LUO Min 1169true in vivo. Therefore, the biodistribution of 99mTc-labeled filamentous phage peptide library via MAG3 (mercaptoacetyltriglycine) were investigated to gain a better understanding of phage circulation in vivo. The experimental results showed that the liver and spleen were the organs of the greatest accumulation, while heart, muscle, pancreas and brain retained less radioactivity. In opposite to other tissues and organs, the radioactivity in stomach, intestine and bone gradually went up with time. The clearance of 99mTc-labeled phage in blood was very fast from 5 min to 30 min and then slowed down. When phage in vivo circulated at enough long period of time, some phage particles could extravasate in some organs or tissues and internalized there. In conclusion, the circulation time of phage in vivo should be experimentally determined beforehand according to the targeted organs and the specific location of target peptides in order to panning a peptide with high specificity and affinity to that target.]]> 2006/12/15 0:00:00 SUN Li-Yan,Chu Tai-Wei and WANG Xiang-Yun 1168true lip3) with the size of 948bp was cloned from the metagenomic DNA by PCR with the primers designed. The deduced Lip3 protein was composed of 315 amino acids (AA) with a molecular mass of 34.577 ku. The motifs GFGNS(GXGXS)and G-N-S-M-G(GXSXG)in the AA sequences of Lip3 were found to be conserved in other lipase. They were most conserved sequence among the serine hydrolase and were necessary for the activity. A 35 ku of Lip3 was purified by Ni-NTA chelating sepharose column from the extract of recombinant E.coli Top 10F′ cell harboring a pLLP-OmpA plasmid inserted with lip3. The purified Lip3 was most active at 25℃ and kept 22% of activity at 0℃. Only 10% of activity was retained after it was incubated at 35℃ for 60 min. The optimal pH value for the Lip3 activity was 8.0. The Km value of the enzyme towards p-nitrophenyl palmitate increased with the increasing of assayed temperature. These results indicated that Lip3 was a typical alkaline cold active enzyme.]]> 2006/12/15 0:00:00 Zhang Jin-Wei and Zeng Run-Ying 1167true 2006/12/15 0:00:00 LI Jing,LI Feng-Bo and WANG Wei 1166true Penicillium chrysogenum, which is important for understanding the industrial fermentation process. PcgstA gene has an open-reading-frame of 840 bp in length, which is interrupted by two introns. The deduced amino acid sequence shows about 50% identity to several characterized filamentous fungi GSTs. The recombinant PcGSTA in Escherichia coli were overexpressed and purified. Enzymatic assays showed that the recombinant PcGSTA had a specific activity with 1-chloro-2, 4-dinitrobenzene of (0.159±0.031) μmol/(min·mg). It was found that the expression level of PcgstA in the penicillin producing medium supplemented with phenylacetic acid, the side chain precursor of penicillin G, was significant down regulated than that in medium without phenylacetic acid. This result suggested that PcGST may be related to phenylacetic acid metabolism in the penicillin producing strain.]]> 2006/12/15 0:00:00 WANG Fu-Qiang,ZHENG Gui-Zhen,ZHAO Ying,REN Zhi-Hong,JIA Qian,HE Jian-Gong and YU Jun 1165true 2006/11/17 0:00:00 TANG Jian-Guang,SHEN Lu,TANG Bei-Sha,ZHANG Yu-Hu,JIANG Hong,LIAO Shu-Sheng,ZHANG Hai-Nan,WANG Chun-Yu,XIA Kun and PAN Qian 1164true 2006/11/17 0:00:00 L?Jun,LUO Liao-Fu,ZHANG Ying and ZHAO Ju-Dong 1163true n=8) were trained on a trace eyeblink conditioning paradigm using a 200-ms tone conditioned stimulus (CS), a 200-ms corneal airpuff unconditioned stimulus (US) and a 600-ms trace interval. Controls consisted of pseudo- conditioned guinea pigs (n=8). Extracellular single unit recordings in vivo were performed in the DG of learner animals during the consolidation of trace eyeblink CRs. The results revealed that all the trace-conditioned animals acquired the trace eyeblink CRs over 14 training days, however, none of the pseudo-conditioned animals did. Furthermore, 23 of 40 single granule cells in the DG of learner animals exhibited heterogeneous activity patterns during the consolidation of trace eyeblink CRs such as increases in activities to the tone CS, trace interval or airpuff US. The results suggested that the DG might participate in the neural circuit important for the consolidation of trace eyeblink CRs, and that the granule cells might encode different information during the consolidation of trace eyeblink CRs.]]> 2006/11/17 0:00:00 HU Bo,LIN Xi,XIONG Yan,HUANG Wei,LIU Guo-Long and SUI Jian-Feng 1162true Verbena officinalis L.(VOL) on mutidrug resistance of JAR/VP16 cells. 4′-methylether-scutellarein produced a synergistic effect combining with the chemotherapeutics and reversed the resistant phenotype of JAR/VP16 cells. TEM analysis and flow cytometry confirmed apoptotic cell features, containing earlier apoptotic modifications, increased apoptotic cell percentage and typical apoptotic peak, when JAR/VP16 cells exposed to 4′-methylether-scutellarein. The gene array results showed that expression levels of some multidrug resistant genes, i.e. MDR1、MRP1、MRP2、MRP6、AHR、COMT and FGF2, were decreased. The expression levels of some apoptosis-promoting genes, i.e. Apaf-1、ASC、ATM、Bad、Bak、Bax and BimL, were increased, whereas those of some apoptosis-suppressing genes, i.e. Bcl-2、BFL1、NAIP and p63, were decreased upon 4′-methylether-scutellarein treatment. Sixteen genes were selected to be detected by real time PCR, and the results were identical with those of gene array. Taken together, 4′-methylether-scutellarein can efficiently reverse multidrug resistance in JAR/VP16 cells. The potential mechanism may be via inhibiting the multidrug resistant gene expressions and/or promoting cell apoptosis.]]> 2006/11/17 0:00:00 XU Shan,LUO Li,ZHU Li-Qun,LI Zhuo,QIU Li-Li,CHEN Qi and XU Chang-Fen 1161true 2006/11/17 0:00:00 YAO Yu-Chang,TIAN Liang,HAN Hong-Bing,CHEN Xue-Hui,LU Ming-Hai,GUO Peng-Cheng,ZHANG Chao-Feng,LI Ning,LIAN Zheng-Xing and LI Wu 1160true P < 0.05, P < 0.01) when auditory deprivation for 7 days. Tone exposure after auditory deprivation for 7 days made NR2B protein level increased significantly (P < 0.05). It showed bidirectional regulation of NMDA receptor subunit NB2B protein. No significant effects of auditory deprivation and tone exposure on the relative expression level of NR2B protein of PND42 (P > 0.05) were found. The results indicated that, auditory deprivation and experience can modify the protein expression level of NMDA receptor subunit NR2B during the critical period of rat postnatal development. The findings provided important data to the study of the mechanisms for the developmental plasticity of sensory functions.]]> 2006/11/17 0:00:00 WU Xiu-Mei,ZHANG Ling,XU Jing-Hong,XU Feng,YANG Wen-Wei,ZHANG Ji-Ping and SUN Xin-De 1159true E. coli. was synthesized, and was expressed in prokaryotic expression pET32a(+) system. The recombinant E. coli was fermented in a small scale and the expression condition was optimized. The expressed recombinant human GIP (rhGIP) was purified by affinity method. The bioactivity of the purified rhGIP was tested through its effect on inhibiting gastric acid secretion and decreasing blood glucose concentration in SD rats. The effect on PC12 cell producing NO free radical was assessed through morphology observation and the NO concentration test. A PC12 cell injury model was constructed by adding Aβ25-35 into the culture medium, different dose of rhGIP was added into this model, and the activity of PC12 was tested by MTT (2-(4,5-dimethylthia-zol-2-yl)-2,5-diphenyltetrazolium bromide) staining. The results suggest that hGIP gene is cloned successfully, the recombinant protein constitutes 35% of the total protein of the cells, some of which was soluble, while the others exist as inclusion body. The relative molecular mass of the recombinant protein is 26 ku as expected. The purified rhGIP showed immunoreactivity. The optimal condition for inducing to expression is with the cell concentration of 0.5 in A600 value, the IPTG concentration of 0.5 mmol/L, under 37℃ and induced for 4 h. The cell lysis supernatant was purified with immobilized metal affinity chromatograph. The production of soluble rhGIP is about 1.2 mg/L culture medium, and the purity is 85%. Purified rhGIP significantly inhibited the secretion of gastric acid and raised the pH value comparing with NC group in SD rats (P < 0.05). There was no significant difference between the rhGIP and standard GIP group (P 0.05).After injection of rhGIP for 15 min, the concentration of blood glucose in plasma decreased remarkably comparing with control group in a high blood glucose environment (P < 0.05). In the later 30 min, there was no remarkable difference (P > 0.05). Also, there was no difference between rhGIP and standard GIP group (P > 0.05).In the research to test the effect of rhGIP on cell nutrition and PC12 cell to avoid neural injury and anoxic injury, it was found that the level of NO was significantly decreased (P < 0.01), the survival rate of the PC12 cells was increased, and neurite extending was better. The activity of the high and medium rhGIP concentration group is higher than that of neural injury group and anoxic injury group (P < 0.05), and the activity is increased as dose-dependence. The differences among rhGIP, GIP and NC group is not remarkable (P > 0.05). The results showed that the rhGIP has been successfully expressed and purified and showed obvious immunoactivity. The recombinant product was shown to have a wide spectrum of biological activity, such as inhibiting gastric acid secretion in the rats stomach, decreasing the concentration of blood sugar in plasma, as well as nutrition and protection to neural cells.]]> 2006/11/17 0:00:00 HU Zhi-Yi,LONG Qi-Xin and WANG Xun-Zhang 1158true In situ hybridization showed that IFRG was highly expressed in granulosa, thecal and cumulus cells, which were consanguineous interrelation to follicles development. So it was proved that IFRG expression was involved in the follicles development, maturation and ovulation in rabbit.]]> 2006/11/17 0:00:00 qibing,HE Xin,LI Wen-Yong and CHEN Qing-Xuan 1157true E. tenella, the relative gene of Rhomboid protein family of E. tenella F2 hybrid strain was amplified by RT-PCR technology and subcloned into the downstream of complex promoter of fowlpox virus vector pUTA2, recombinant fowlpox virus transfer vector was constructed successfully. Plasmid of fowlpox recombinant virus and fowlpox virus were co-transfected into chicken embryo fibroblast (CEF) and identified by BrdU drug, RT-PCR and Western blotting respectively. The result showed that the E. tenella fowlpox recombinant virus (rFPV-Rhomboid) was obtained. Furthermore, rFPV-Rhomboid was amplified in CEF, then chicken was immuned with rFPV-Rhomboid and the indication of immunity was detected. The result indicated that the contents of CD4+ and CD8+ in peripheral blood were significantly higher than that in control group (P<0.05). The weight gained of chicken vaccinated rFPV-Rhomboid was different from the chickens of control group significantly (P < 0.05). The recombinant protein had immunogenicity and could induced strong immune response and immune protection ability.]]> 2006/11/17 0:00:00 YANG Gui-Lian,lijianhua,zhangxichen,zhaoquan and YIN Ji-Gang 1156true Glycine max) N2899 seed germination at specific stages of 0 h, 8 h, 36 h and 60 h after imbibition. The results show that on the 2-DE gels stained by coomassie brilliant blue, PDQuest image software detected about 350 protein spots, of which 24 spots show more than 2.5-fold changes in abundance, and most of soybean seed storage proteins weren't mobilized during seed germination. At the first stage of germination, 10 proteins show changes in abundance. At the second stage, the quantities and categories of differentially expressed proteins increased and the abundance of 15 proteins dynamically changed. During radicle protrusion seed coat, the abundance of 14 proteins reached a peak. These suggested that the internal metabolisms of imbibition seeds became more active. These 24 proteins treated by tryptic in-gel digestion were characterized by matrix-assisted laser-desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) and peptide mass fingerprintings of all were obtained. Soybean UniGene database was used to identify proteins, of which 6 proteins were identified. They were nucleotide diphosphate kinase, proglycinin A1aB1b subunit, thioredoxin fold, 35 ku seed maturation protein, heat shock protein and seed maturation protein PM36. The potential functions of these proteins during seed germination were discussed.]]> 2006/11/17 0:00:00 XU Xiao-Yan,ZHENG Rui,LI Chun-Mei,GAI Jun-Yi and YU De-Yue 1155true Glechoma hederacea agglutinin (Gleheda) is a novel glycosylated lectin isolated from the leaves of G. hederacea. Like other glycosylated proteins, the detection of Gleheda by immunological methods is often hampered by the cross-reactivity of the polyclonal antibodies with unrelated glycoproteins. Hence a protocol to purify monospecific polyclonal antibodies from a crude antiserum raised against Gleheda was developed. After selective ammonium sulfate precipitation and successive affinity chromatography on columns of Sepharose 4B with immobilized Gleheda and Robinia pseudoacacia agglutinin (RPA), respectively, ion-exchange chromatography on a column of Q Fast Flow was used for further purification. The specificity of the antibody fractions from each step was tested by double immunodiffusion assay and analyzed by Western blot. Results revealed that affinity chromatography of the immunoglobulin fraction on the immobilized Gleheda antigen yielded an antibody preparation that still cross-reacted with many proteins in leaf extracts. Depletion of nonspecific cross-reacting antibodies directed against the glycan part of the glycoprotein by affinity chromatography on immobilized RPA removed most but not all nonspecifically reacting antibodies. Only upon further purification by ion exchange chromatography an IgG fraction of monospecific antibodies that reacted exclusively with Gleheda could be obtained and accordingly was suitable for immunodetection studies. This antibody purification procedure promises simplicity and efficiency. In addition, this method does not require expensive facilities.]]> 2006/11/17 0:00:00 WANG Wei-Fang,VAN DAMME Els and HE Chao-Zu 1154true z, αA, αB, αC) and two loops (ZA loop，BC loop). Alignment of the amino acid sequence of BRD7 protein with known structure of proteins in PDB (protein database), it was considered that Bromodomain of BRD7 has the same characteristic. Based on results of bio-informatics, mutants of Bromodomain were constructed. The peptide binding assays was performed to detect the binding of mutants with acetylated histone H3. The results indicated that two loops (ZA loop，BC loop) were essential for Bromodomain binding to acetylated histone H3, while two loops(ZA loop，BC loop) could influence the sub-cellular location of BRD7 through indirect immunofluorescence. At last, it was found that the CBP(CREB binding protein) is a novel interaction protein of BRD7. The CBP not only has histone acetyltransferases (HATs) to acetylate lysines on N-terminal tails of histones but also plays a key role as co-activator for a wide variety of transcription factors.]]> 2006/10/18 0:00:00 peng cong,Li xiao ling,LI Sia-Yu,liu hua ying,zhou ming,Zhang li ming,Luo xiao ming,Sheng shou rong and Li Gui Yuan 1153true 2006/10/18 0:00:00 Sun dongmei,LIU Zhong-Bo,ZHAO Yan,GONG Zhen-Wei,LI Dan,WANG Xi-Yuan,ZENG Xian-Lu and liu wenguang 1152true in vivo estrogen hormones. It has been known that estrogenic endocrine disrupters, which can cause intersexuality and reproduction decline in animals and human, can also induce synthesis of female-specific proteins such as vitellogenin (vtg) in male and juvenile fish. To establish a novel in vivo test system for rapidly detection of estrogenic chemicals, a transgenic zebrafish has been developed. First, the expression vector pvtg1-EGFP by ligating a 1.7 kb zebrafish vtg1 promoter to a reporter gene EGFP (enhanced green fluorescent protein) was constructed. Next, the pvtg1-EGFP DNA was microinjected into one-cell stage embryos of the zebrafish. Then the embryos were raised and divided into three groups: group one (blank control) remained in embryo media; group two (solvent control) and group three were subject to continuous exposure of 0.001% alcohol and 1ng/L 17α-ethinylestradiol (EE2), respectively. The three groups were observed for GFP expression with fluorescent microscopy every 12 h. Green fluorescence was first observed on day 5 in the livers of group three larvae, while it was observed neither in group one nor group two. Furthermore, RT-PCR and whole in situ hybridization experiments were performed and concurrent accumulation of vtg1 and EGFP mRNAs in larvae after EE2 exposure were observed, which indicated that the expression of EGFP transgene was driven by the vtg1 promoter. Last, by amplifying EGFP gene from the genomic DNA of fluorescent fish, it was verified that the EGFP transgene had been inserted into the genomic DNA of these fish. Through observing green fluorescence in these transgenic fish, the presence of estrogenic substance in the water can be easily detected. The transgenic zebrafish offers an easy to use tool for effective monitoring of environmental estrogens.]]> 2006/10/18 0:00:00 Chen Hao,Yang Jian,Wang Yuexiang,Jiang Qiu,Xu Hui and Song Houyan 1151true In vitro neurosphere was cultured in a serum-free defined medium DMEM/F12 supplemented with bFGF and EGF. Dissociated cells were plated onto poly-d-lysine-coated coverslips and propagated in medium containing recombined Wnt3a-adenovirus. Plenty of Nurr1 were detected by RT-PCR after 3 days. Wnt3a combined AA would improve NSC differentiation into dopaminergic (DA) neuron. The quantity of DA neuron is obviously more than the AA alone group's. Moreover, the expression of TH mRNA is 1.86 fold in Wnt3a combined AA group. Induced cells were immunostained for TH and DAT. The proportion of TH-positive was (37.42±2.54) % (P<0.05) in the AA group , accordingly in Wnt3a combined AA group it was (73.96±2.61) % (P<0.05). These results indicate that Wnt3a promoted the proliferation of precursor cells expressing the orphan nuclear receptor-related factor 1 (Nurr1) . Large numbers of DA neurons are obtained by adding AA in the culture of NSC modified Wnt3a in vitro. Therefore, the successful conversion of neural stem cells in vitro resulting in the desired dopaminergic neuronal phenotype, could provide a solution to the problem of limited availability for clinical surgical transplantation therapies, which are currently in progress for the treatment of neurodegenerative diseases such as Parkinson's disease. These findings indicate that Wnt3a is key regulators of proliferation and differentiation of DA precursors.]]> 2006/10/18 0:00:00 han shu,SHI Wei,LI Yan-Hua,YAO Hai-Lei,XIE Xiao-Yan,CHEN Lin,SHI Shuang-Shuang,BAI Ci-Xian,NAN Xue,YAN Fang,WANG Yun-Fang and PEI Xue-Tao 1150true Helicobacter pylori (Hp) recombinant protein vaccines, the oral lipsome-encapsulated vaccines with/without CT were used. Phosphatidyl choline (PC) and cholesterols (Chol) were prepared using reverse evaporation method whose size distribution of folate liposomes was measured by electronic microscopy. BALB/c mice were divided into six groups and immunized by BPS alone, liposome alone, rUreB plus CT, liposome-encapsulated rUreB, liposome-encapsulated rUreB plus CT and liposome-encapsulated (rUreB and rKat) plus CT orally respectively once a week for four weeks. All mice were challenged by alive Hp three times in two weeks after the last immunization and sacrificed in five weeks after the last challenge. Hp was determined by the fast urease test. The bactrerial colonizing density semi-quantitation, the inflammation severity grades and activity grades of the gastric histopathology were observed. IFN-γ and IL-4 mRNA expression of spleen T lymphocyte cell in different groups were studied with RT-PCR. The lipsome-encapsulated vaccines were obtained successfully. The mean size of folate liposome was (0.7±0.2) μm. Protective rates of BPS alone, liposome alone, rUreB plus CT, liposome-encapsulated rUreB, liposome-encapsulated rUreB plus CT and liposome-encapsulated (rUreB and rKat) plus CT were 0(0/11), 0(0/11), 58.3%(7/12), 54.5%(6/11), 63.6%(7/11) and 75.0%(9/12) respectively. Protective rate of rUreB plus CT was equal that of the liposome-encapsulated rUreB group (P>0.05). Protective rate of liposome-encapsulated (rUreB and rKat) plus CT was significantly higher than that of liposome-encapsulated rUreB plus CT (P<0.05). The mice gastrically inoculated with the latter four immune antigens had been significantly less infected by Hp and had mild inflammation of gastric tissue (P<0.05). There was not significant difference among any vaccine group (P>0.05). After Hp attacked, the level of INF-γ mRNA expression of every vaccine group was significantly lower than that of the PBS and liposome group (P<0.05) while the level of IL-4 mRNA expression of every vaccine group was significant higher than that of the PBS and liposome group (P<0.05). There was not significant difference among any vaccine group (P>0.05). The lipsome-encapsulated vaccines can significantly decrease Hp coloning density and reduce inflammation degree in mouse model. The double- candidate vaccines have more stronger immune protective function against Hp than that of sigle-candidate vaccines. After Hp attack, the mice which infected without vaccine induce a predominant Th1 immune inflammatin response while the mice which have accepted vaccine can induce a predominant Th2 protective immune response.]]> 2006/10/18 0:00:00 Huang Wen,LI Zhao-Shen,BAI Yang,WANG Ji-De,ZHANG Ya-Li and ZHOU Dian-Yuan 1149true Streptomyces conjugative linear plasmids, containing a free 3′-end but a protein-capped 5′-end, can potentially undergo cell-to-cell transfer by transfer of non-nicked DNA. It was reported that circular derivatives of the Streptomyces lividans linear plasmid SLP2, as well as the parental linear plasmid itself can transfer efficiently. And the genetic requirements for such transfer was described. Efficient transfer of plasmid requires six co-transcribed SLP2 genes, encoding a Tra-like DNA translocase, cell wall hydrolase, two cell membrane proteins that interact with an ATP binding protein, and a protein of unknown function. Reduced transfer efficiency of plasmid from SalⅠR-/M- to SalⅠR/M hosts argues that transfer of both the circular and linear forms of the plasmid involves double-stranded DNA. These results suggest that conjugal transfer occurs by a similar mechanism for SLP2-derived linear and circular plasmids, and cellular membrane/wall functions in the transfer process.]]> 2006/10/18 0:00:00 Xu Ming-Xuan,Zhu Ying-Min,Shen Mei-Juan,Jiang Wei-Hong,Zhao Guo－Ping and Qin Zhong-Jun 1148true in vitro fertilized (IVF) blastocysts. Hypomethylation of the Xist gene was observed in the cloned blastocysts (43% and 17% in FFB and FOV derived blastocysts, respectively), while the Xist gene in 49% IVF blastocysts appeared to be methylated. On the contrast, somatic donor cells showed high percentage of methylation (66% and 63% in FFB and FOV cells, respectively). These results suggest that the DNA methylations of the Xist gene were reprogrammable and the detected CpG islands may regulate expression of the Xist gene. Altogether with the previous data, It was shown that development rate of cloned blastocysts derived from FFB is lower than that from FOV of the same origin, while pregnancy and birth rates appear to be opposite. It can be proposed that cloned blastocysts derived from different types of somatic cells differ in DNA reprogramming that may affect embryo and fetal development.]]> 2006/10/18 0:00:00 Cao Geng-Sheng,GAO Yu,DAI Yun-Ping,LI Rong,ZHAO Yao-Feng and Li Ning 1147true in vitro system for promoter-independent assembly of a functional mammalian RNA polymeraseⅡ elongation complex using highly purified polymerases and synthetic RNA and DNA oligonucleotides was presented. It was shown that the 9-nucleotide RNA∶DNA template hybrid is necessary and sufficient for the formation of a stable elongation complex with human RNA polymeraseⅡ, while further inclusion of the non-template DNA strand to form the complete transcription “bubble” actually destabilizes the already formed RNA:DNA:RNA polymeraseⅡ complex. In addition, by introducing a DNA damage at a specific site in the template DNA, this assay can potentially be used for characterizing the processes of transcription-coupled repair of DNA damages.]]> 2006/10/18 0:00:00 LIU Wei-Feng and SVEJSTRUP Q JESPER 1146true 2006/10/18 0:00:00 WEI Ling,LIU Xuan and CAO Cheng 1145true 441 was phosphorylated by recombinant GSK-3β in vitro, and the site-specific phosphorylation was detected by Western blots using site-specific and phosphorylation-dependent tau antibodies. The kinetics of phosphorylation by GSK-3β to total tau and to individual site was studied by incubating GSK-3β with various concentration of tau at 30℃ for 10 min. These velocities of phosphorylation reaction were determined at various concentrations of tau by measuring 32P incorporation to tau and phosphorylation level at individual site detected with immuno-dot blots using site-specific and phosphorylation dependent anti-tau antibodies, respectively. The Km values of GSK-3β toward total tau and toward individual site were calculated by using Lineweaver-Burk double-reciprocal method. The site-specific phosphorylation of tau by GSK-3β was further confirmed in cultured CHO cells by co-transfection of tau441 with GSK-3β. It was found that GSK-3β phosphorylated tau at several sites, including Thr181, Ser199, Ser202, Thr205, Thr212, Thr 217, Thr231, Ser396, and Ser404. The Km value of GSK-3β toward total tau was 42 μmol/L, but the Km value of GSK-3β to every site was different. Among all the phosphorylation sites detected here, Ser396 phosphorylation catalyzed by GSK-3β showed the lowest Km, only 16 μmol/L. In cultured CHO cells, GSK-3β over-expression also induced tau phosphorylation at Ser396 the most dramatically. These results suggested that GSK-3β catalyzed tau phosphorylation at several sites with different efficiency, and Ser396 was the most efficient site for phosphorylation by GSK-3β .]]> 2007/6/13 0:00:00 LIU Fei,SHI Jian-Hua,DING Shao-Hong and YIN Xiao-Min 1144true 2007/4/24 0:00:00 LIU Qing-Bin,LI Yan-Hua,YANG Yin-Xiang,YUAN Hong-Feng,ZHANG Rui,QIN Li-Peng,LU Yang,ZHANG Sai-Nan,SUN Di-Jin,CHEN Lin,BAI Ci-Xian,NAN Xue,WANG Yun-Fang and PEI Xue-Tao 1143true M-T, recombination expression plasmid PET-32a(+)-wzy1-1 was constructed according to recombination of cloned vector PUCM-T-WZY1-1 of wheat dehydrin gene in this experiment. Then the recombination was transformed into the host strain E. coli BL21(DE3), was induced by IPTG and target protein was got. The expression production was detected by SDS-PAGE, and Western blot assays revealed that the fusion protein was expressed in soluble form with a relative molecular mass 37 ku, and the fusion protein was expressed at high level. After purification by Ni-NTA resin affinity chromatography and electroelution in bagfilter, the fusion protein was used to induce the production of polyclonal antibody in rabbits and ELISA detection showed the antigenicity of the fusion protein was satisfactory, Western blot showed the antiserum raised against the recombination dehydrin protein in rabbits could react to the protein expressed specifically, and the protein of zheng yin 1# wheat under drought stress was extracted, through the SDS-PAGE, Western blot assays revealed that there is a protein band with a relative molecular mass 28 ku, this revealed that antiserum could react to the dehydrin protein expressed in wheat leaf specifically and demonstrated its good antigenicity.]]> 2007/3/20 0:00:00 YANG Ying,ZHANG Lin-Sheng,ZHANG Xiao-Juan and SHAN Lun 1142true IC50=(54.56±5.18) μmol/L, IC50=(28.29±1.32) μmol/L, respectively). These heptapeptides could be used for developing new anti-HIV drug candidates, as well as for structural studies of the three-dimensional structure of the entire integrase molecule.]]> 2007/4/11 0:00:00 ZOU Yuan and ZHAN Jin-Biao 1141true 2007/4/11 0:00:00 Sun Dong-Bo,LANG Hong-Wu,SHI Hong-Yan,CHEN Jian-Fei,CUI Xiao-Chen,WANG Cheng-Bao,TONG You-En and Feng Li 1140true 2+ is a ubiquitous second messenger which plays a key role in early development of embryos. Ca2+ probes (Fluo-4 or Indo-1) were injected into zebrafish eggs to detect the distribution of free Ca2+ during their first cleavage using confocal microscopic or dual-wavelength ratiometric imaging. A high Ca2+ zone was first observed in the animal pole right before the first cleavage, then it extended along the cleavage furrow and the Ca2+ signal remained high in this region throughout the first cleavage. Intracellular Ca2+ concentration ([Ca]i) was measured via Indo-1 dual-wavelength system, and it was shown to be homogeneous within the whole embryo before the first cleavage. During the first cleavage, [Ca]i increased significantly near the cleavage furrow, while it remained unchanged in other areas. As the dual-wavelength ratiometric imaging eliminates the artifacts due to indicator inhomogeneity, the results provided an unequivocal quantification for the Ca2+ dynamics associated with the first cleavage of embryonic development.]]> 2007/5/30 0:00:00 GAO Wen-Xue,ZHOU Peng,WEI Wei,WEN Lu,ZHANG Bo and WANG Shi-Qiang 1139true in vitro with GFP-DNA (green fluorescent) triplex micelles. Cell culture studies revealed that DAC triplex micelles markedly increased the level of gene transferred to A10 cells, with more than 4-fold increase in transfection compared to DC group without anti DNA antibody and 11-fold increase compared to DA group without cationic lipid. Fluorescent DNA studies demonstrated greater cell uptake in vitro with DAC compared to the control formulations. Confocal microscopy studies confirmed nuclear entry in A10 cells with DAC, while formulations without anti-DNA antibody demonstrated no nuclear entry. It can be concluded that plasmid DNA-anti DNA antibody-cationic lipid (DAC) triplex micelle was a novel non-viral gene vector with high transfection efficiency and no cytotoxicity. It was hypothesized that DAC triplex micelles could enhance DNA delivery through mechanisms involving: increased levels of DNA incorporation into micelles due to antibody binding, enhanced plasmid DNA entry into nuclear due to the nuclear entry properties of anti-DNA antibody, and increased transfection efficiency via co-operation of anti-DNA antibody and cationic lipid. Gene delivery using DAC triplex micelles should be suitable for a wide array of single or multiple therapeutic gene strategies and for further successful gene therapy.]]> 2007/8/13 0:00:00 Song Cun-xian,Zhang Lin-Hua,Zhang Chao and LEVY RJ 1138true in vitro or ex vivo gene delivery because of their relatively low titers using transient transfection experiments. In order to develop an efficient transient transfection method for large-scale production of high titer lentiviral vector stocks, a minimal lentiviral vector producing system based on vaccinia virus that synthesizes T7 RNA polymerase was developed. BHK21 was co-transfected by three main plasmids containing the transducing plasmid pVECRNA, the packaging plasmid pGAGPOL and the envelope plasmid pVSVG, and thereafter infected with the vaccinia vTF-3 containing bacteriophage T7 RNA polymerase gene using Lipofectamin2000TM. After 4 days, the culture supernatant of lentiviral vectors was collected, the RNA from the supernatant was examined by the RT-PCR, the protein from the supernatant was examined by Western blot, and the supernatant was used to transfect normal 293T , HepG2 and Vero, which were observed by the immunofluorescence microscopy. The type of cell lines, plasmids dosage and the MOI (the proportion between cell numbers and virus copies) were considered so critical to the output of this system that 3×3×3 factorial design was used to explore the yield optimization of this system. As judged by the results of RT-PCR and the Western blot, lentiviral vectors were found in the culture supernatant; as judged by immunofluorescence with microscopy, 293T, HepG2 and Vero which were transfected by the supnantant expressed the report protein - green fluorescent protein(GFP), the results confirmed the valid infectivity of the lentiviral vector produced by the system. Eventually, the best titers of lentiviral vector stocks was up to 1.3×108 tu/ml, which is one order of magnitude higher than the output of classical manufacture system. The new poxviral/lentiviral hybrid system for efficient lentiviral vector production was initially established. It provides the basis for the future development of industrial application.]]> 2007/8/13 0:00:00 MA Qiang,LI Ming,DONG Wen-Qi and WU Ying-Song 1137true E.coli. The Gram-positive pathogenic bacterium Enterococcus faecalis has a fatty acid composition very similar to that of E.coli. Bioinformatic analysis reveals that though E. faecalis has two fabF homologues, there is no recognizable fabB homologue in the genome of E. faecalis. Two fabF homologues (fabF1 and fabF2) were amplified by using E. faecalis V583 genomitic DNA as template, and two plasmids, pHW13 (fabF1) and pHW14 (fabF2), were constructed. The results of experiments in vivo and in vitro have shown that fabF1 gene could complement E. coli fabB mutation and FabF1 possessed β- ketoacyl-acyl carrier protein synthase Ⅰ(FabB) activity, while fabF2 gene could complement E. coli fabF mutation and FabF2 had β- ketoacyl-acyl carrier protein synthase Ⅱ(FabF) activity. Meanwhile the data also shown that FabF2 possessed partial function of β-ketoacyl-acyl carrier protein synthase Ⅰ(FabB), and it could make E. coli fabB mutation synthesized low amount of unsaturated fatty acid. From these data it is clear that FabF species enzymes could have activity of β- ketoacyl-acyl carrier protein synthase Ⅰ(FabB).]]> 2007/8/13 0:00:00 WANG Yu-Qi,SUN Yi-Rong,CHEN Yi-Cai and WANG Hai-Hong 1136true P < 0.05), SOD activity were higher, CAT activity was decreased remarkably in pathological scars than that in normal skin tissues (P < 0.05). And the activity ratios of CAT/CuZn-SOD and GPX/CuZn-SOD were obviously reduced (P < 0.05). However, no significantly differences were observed in the decrease activity of GPX. Differences in above-mentioned indexes all were not remarkable between hypertrophic scars and keloids. These data suggest that the changes of XO, CuZn-SOD, CAT and GPX activities may be a reason resulted in an increase in the level of reactive oxygen species in pathological scars. In the alternative of antioxidants, CAT may be more reasonable.]]> 2007/5/8 0:00:00 LI Wei-Ren,CEN Ying,LIU Xiao-Xue,LI Xiao-Hong and ZUO Feng-Qiong 1135true 2007/6/11 0:00:00 SUN Cheng-Rong,TANG Jian-Wu,SUN Ming-Zhong,LIU Shu-Qing,ZHANG Hong-Ying,WANG Bo,Song Bo,Zhang Ya-Nan,Zhang Zhu-Qing and ZHAO Zhi-Ying 1134true in vivo function of mouse FSP27. The double strand siRNA of mouse FSP27 corresponding to nucleotides 270 to 291 was synthesized and inserted into pSilencer2.1. pSilencer-siFSP27 was co-transfected into 293T cells with the HA-mFSP27 expression vector to test its knock-down efficiency. The FSP27 siRNA was then transferred to a lentiviral vector. Lentivirus were generated and used to infect 3T3-L1 cells. It was shown here that lentivirus containing FSP27siRNA can effectively knockdown FSP27 expression in 3T3-L1 cells. Establishment of FSP27 knock-down cell line provides a useful tool for the study of in vivo function FSP27.]]> 2007/8/13 0:00:00 YAO Hui-lan and LI Peng 1133true 2007/8/13 0:00:00 RUAN Xiao-Gang,WANG Jin-Lian and LI Hui 1132true 2007/5/16 0:00:00 LI Shu-Fang,ZHOU Ming,LIU Hua-Ying,XU Xiao-Jie,PENG Cong,PENG Shu-Ping,WU Ming-Hua,LI Xiao-Ling and LI Gui-Yuan 1131true Nicotiana glutinosa plants, in terms of chloroplast structure and photosynthesis, a mutant Fny-CMVΔ2bpro, which cannot express the 2b protein, was achieved by introducing mutant sites in the 2b gene of Fny-CMV. N. glutinosa seedlings were inoculated with wild-type Fny-CMV and the mutant Fny-CMVΔ2bpro, and were analyzed for symptom expression, chlorophyll content, photosynthetic rate, and ultra-structural alteration of chloroplast. Up to 30 days post inoculation, wild-type Fny-CMV caused symptoms of severe mosaic, leaf deformation, and stunting, reduced photosynthetic rate and chlorophyll content, and altered the ultra-structure and morphological characters of the chloroplasts. However, host seedlings inoculated with the mutant Fny-CMVΔ2bpro expressed only slight mosaic symptom. Their photosynthetic rates and chlorophyll contents were not significantly different from those of the mock-inoculated plants, and the ultra-structure and morphological characters of their chloroplasts appeared to be normal. The observed low photosynthetic rates and chlorophyll contents were related to the breakage of the chloroplast morphology and ultra-structure. Results of Northern blotting showed that the virulence of 2b protein was associated with high accumulation level of CMV progeny RNAs in systemic leaves. Non-expression of the 2b protein reduced the accumulation levels of its genomic RNAs 1 and 2. The level of subgenomic RNA4, encoding CP protein, was found to be significantly decreased.]]> 2007/4/24 0:00:00 CHEN Fei-Fei,DU Zhi-You,LIU Xin,XIE Li and CHEN Ji-Shuang 1130true in vivo in a nude mice xenograft model. It was demonstrated that adenovirus-mediated ODC and AdoMetDC antisense expression could inhibit tumor cell growth, lead to cell apoptosis and reduce tumor cell invasiveness. Polyamine levels were significantly decreased in Ad-ODC-AdoMetDCas-treated cells compared with controls. This adenovirus also induced tumor regression in established tumors in nude mice. It was suggested that as a new anticancer reagent, the recombinant adenovirus Ad-ODC-AdoMetDCas holds promising hope for the therapy of lung cancers.]]> 2007/5/8 0:00:00 Tian Hui,LIU Xian-Xi,ZHANG Bing,SUN Qi-Feng and SUN Dong-Feng 1129true 2+, which were loaded with (15 μl) Fluo-3 AM , were detected by confocal laser scanning microscope(CLSM). The signaling inhibitors, such as SB203580(p38MAPK specific inhibitor), PD98059(MEK-1/2MAPK specific inhibitor), LY294002(PI3Ks specific inhibitor), cytochalasin B(microfilament specific inhibitor), EGTA (Ca2+ specific inhibitor), were used to investigate mechanical signaling pathway by (3% 0.5Hz) Cyclic strain. The results indicated that 3% strain stimulus could induce increasement of [Ca2+]i - dependent fluorescence at 2 min. Its fluorescence intensity were 143.68,which were significantly different compared with 1,3,5,10min groups and 1% and 10% strain groups (P < 0.01). The increasement of Ca2+ levels were delayed upto 10 min, when the cells were treated with CB. EGTA (5 mmol/L) could inhibited strain induction increasment of Ca2+ levels. Strain (3%, 0.5 Hz) could induce Ca2+ spark event in D1 cells at 2 min, however the cells were treated with CB Ca2+ spark event were delayed until 5min. The expression of OCN and OSX mRNA were completely inhibited by five specific inhabitors in unstrained condition. Inhibition of ERK1/2 and p38 pathway promoted the expression of OSX and OCN mRNA by strain.The expression of OSX and OCN mRNA to inhibit PI3Ks pathway were partly activated by strain. Application of strain could not activate OSX and OCN mRNA expression when extracellular Ca2+ greatly inhibited with EGTA (5 mmol/L) and microfilament were damaged with CB.These results demonstrate that mechanical signals regulate MSCs function, suggested Ca2+ signaling, PI3Ks pathway and microfilament play an important role in transduction mechanical signal in MSCs differentiation.]]> 2007/6/22 0:00:00 ZHAO Hong-Bin,LU Tong-De,MA Jing,MA Hui and ZHANG Xi-Zheng 1128true Escherichia coli K12, Borrelia burgdorferi, Bacillus cereus ZK). According to our previous work, codon use frequency (FCU) was selected as the sequence feature, in respect that it is inherently the fusion of both codon usage bias and amino acid composition signals. In addition, another computational method was proposed considering strand asymmetry and predicting horizontal gene transfers of leading strand and lagging strand of genomes under consideration, respectively. To avoid the occasionality of simulating gene transfer through artificially inserting phage genes, 100 times of the transfer-and-recover experiment were repeated and arithmetic average of measurement for each genome being considered were reported to evaluate algorithm's performance. Ten-fold cross-validation was used for both parameter and accuracy estimation. The best results were obtained for C-Support Vector Classification (C-SVC) type by using the radial basis function kernel with γ=100, while for one-class SVM type the best performance was obtained using the polynomial kernel of three degree. The performance of the approach was compared with that of Tsirigos' method ,which is one of the best predictive approachs to date in detecting of horizontal transfer genes. Firstly, for the original method that did not consider the strand asymmetry, the C-SVC type has a high relative improvement(RI) of 31.47% on hit ratio for Escherichia coli K12, while the one-class SVM type has RI of 11.61% for Borrelia burgdorferi. Moreover, as theoretically expected, the method considering the strand asymmetry resulted in higher RI than the original method. In order to examine the approach's performance in detecting factual gene transfer events, the approach was applied in genome of Enterococcus faecalis V583. It is not only succeed in recovering all the seven factual horizontally transferred genes, also found that the whole segment from 7 kb upstream of gene EF2293 to 38 kb downstream of gene EF2299 was probably transferred into E. faecalis V583 genome simultaneously with the above seven genes.]]> 2007/5/23 0:00:00 WU Jian-Sheng,XIE Jian-Ming,ZHOU Tong,WENG Jian-Hong and SUN Xiao 1127true 2007/5/15 0:00:00 LUO Ye,WANG Lan,GAO Xia,DENG Wei-Wei,YU Peng,ZHANG Chen-Ying,LU Yang,HAO Yu and SHI Tai-Ping 1126true in vitro migration and invasion assay, apoptosis ratio was detected by flow cytometry(FCM) and TUNEL method, prominent altered signal pathway was analyzed through a Human Q Series Signal Transduction in Cancer Gene Array. The results showed that the up-regulation of HSP27 expression in HCC cell lines was consistent with metastasis potentials. Using RNA interference technique, mRNA and protein level of HSP27 in MHCC97H cell line were specifically reduced by up to 80%. In vitro migration and invasion assay showed the average migratory and invaded cell numbers per field were 21.36 ± 2.92, 19.88 ± 2.23, 11.40 ± 2.05 and 26.35 ± 3.29, 24.43 ± 3.17, 10.92 ± 2.27 respectively in MHCC97H, control RNAi, RNAi groups. Moreover, MHCC97H cell apoptosis ration was 15.12%, 17.56%, 27.64% (MHCC97H, control RNAi, RNAi) respectively by FCM analysis, which was reconfirmed by TUNEL morphological assessment. Furthermore cDNA microarray analysis revealed that NF-κB pathway activation was inhibited after HSP27 RNAi. Immunoblotting analysis showed that the decreasing of nuclear activated NF-κB p65 and phosphorylated IκBα, meanwhile reducing of the association between IKKβ and IKKα in MHCC97H cells after HSP27 RNAi, further co-immunoprecipitation assay showed interaction of IKKβ, IκBα with HSP27 in three HCC cell lines. Altogether, these findings revealed possible roles of HSP27 in being of HCC cell lines metastatic potentials through involving in cellular NF-κB pathway activation.]]> 2007/3/20 0:00:00 GUO Kun,LIU Yin-Kun,ZHOU Hai-Jun,DAI Zhi,CHEN Jie,SUN Rui-xia,SUN Qiang-Lin,LU Wen-Jing and KANG Xiao-Nan 1125true P < 0.01) and spleens (1.1, P < 0.01) compared with the saline control. In addition, this approach also have much better protection than that vaccination with combined DNA or BCG alone (P < 0.05). The elevated CD4+ and CD8+ T cells percentage (P < 0.05) in PBMC and higher IFN-γ concentration (1250 ng/L, P < 0.01), IL-2 concentration (230 ng/L, P < 0.05) stimulated by Ag85B protein in spleen cells supernatant in BCG boosted mice 21 days after third vaccination, indicating that the DNA prime-BCG boost strategy significantly enhanced the Th1 type cell response, which is correlated with the protective efficacy against tuberculosis. Furthermore, immuno-histochemistry assay showed that there were more Perforin expression cells, secreted mainly by CD8+ T cells, in the lung tissue of the mice primed with DNA prior to BCG. Taken together, the heterogonous boost combination provided superior protection by stronger CD4+ (Th1) and CD8+T cell mediated immune response, which suggested that it will be a promising regimen for MTB vaccine development.]]> 2007/5/31 0:00:00 LI Min,YU Da-Hai and CAI Hong 1124true +vEGFR-2+, the special markers of EPCs. If the both markers of this part was positive, the rest part of the cells was continuously cultured for passaging. This method was named as “Micropore-Method”. The conspicuous stem cells colony-forming units were observed under microscope after four days in cluture, about 7% CFUs were CD133+/VEGFR-2+ positive EPCs-CFUs. After further culturing 7 days, Purity of cells with special markers of CD133+/VEGFR-2+/CD34+ was over 70% identified by flow cytometry. Passaging cells can form capillary tube like formation and differentiate to endothelial like cells expressing EC special marker vWF. It can be concluded that “Micropore-Method” is a new successful way to isolate EPCs from rat bone marrow.]]> 2007/5/24 0:00:00 WANG Zuo,TONG Zhong-Yi,ZHOU Xiao-Feng,JIANG Zhi-Sheng,TANG Chao-Ke,SONG Yan-Ming and TIAN Yong-Feng 1123true 2007/4/13 0:00:00 Sun Yi,YI Hong,YANG Yi-Xuan,ZHANG Peng-Fei,LI Mao-Yu,LI Jian-Ling,Yang Fang,Xiao Zhi-Qiang and Chen Zhu-Chu 1122true cis/trans isomerization of Xaa-Pro motif in the peptide and protein plays an important role to influence their conformation and function. Here, a series of model peptides including phosphorylated and its unphosphorylated counterparts were designed and synthesized. Preliminary 1H NMR experiments and molecular dynamics (MD) simulation were used to analyze the peptidyl-prolyl cis/trans imide bond isomerization. The data indicated that the side-chain O-phosphorylation of the Xaa residues preceding proline affected evidently the isomerization and thereby regulated the peptides conformations. The charges of the phosphate moiety as well as their steric effects might be the driving force for the conformational changes of these phosphopeptides. Moreover, the obtained most stable multiple configurations and their statistic cis/trans concentration distribution in MD simulation were basically consistent with the NMR experiments, which demonstrated that phosphorylation increased the cis conformation of the peptide and the maximum cis ratio is given while the phosphate group has no negative charge.]]> 2007/5/28 0:00:00 ZHU Zhen-Tai,SUN Ming,GUO Yan-Ting and LI Yan-Mei 1121true squint mRNA, MZoep mutant embryos that are deficient in Nodal signaling, and wildtype embryos at the 30% epiboly stage. Transcripts (genes) with at least two-fold changes in expression level between wildtype and the other samples were identified. In squint mRNA-injected embryos, 265 transcripts show an increased expression level and 111 have a decreased expression level; in MZoep embryos, the expression of 1 495 transcripts increases while 550 transcripts express at a decreased level. Furthermore, overexpression of squint causes increased expression of 26 and decreased expression of 11 annotated transcription factor genes; in MZoep embryos, the number of transcription factor genes showing an increase and decrease of expression are 69 and 30, respectively. These results would provide useful information for further studying mechanisms and biological functions of Nodal signal transduction.]]> 2007/5/15 0:00:00 SUN Zhi-Hui and MENG An-Ming 1120true 2007/4/23 0:00:00 YANG Ji-Yun,ZHANG Si-Zhong,GUO Hong,ZENG Xiang-Yuan and MA Bu-Ren 1119true P < 0.05). The similar results have been observed in foam cells treated with FRK. ApoA-Ⅰ, in combination with FRK, contributed to a much larger increase in protein expression of ABCA1 (80%, P < 0.05) and cholesterol efflux (36%, P < 0.05) from THP-1 macrophage derived foam cells. In converse, treatment THP-1 macrophage derived foam cells with apoA-Ⅰ and SQ-22536 markedly down-regulated protein expression of ABCA1 (26.7%, P < 0.05) and decreased cholesterol efflux (22.1%, P < 0.05). However, apoA-Ⅰ, FRK and SQ-22536 alone or combination did not influence mRNA expression of ABCA1. These findings suggest that apoA-Ⅰ may stabilize ABCA1 protein, and then increase cellular cholesterol efflux through PKA signaling.]]> 2007/5/22 0:00:00 YANG Jun-Hao,DAI Xiao-Yan,OU Xiang,HAO Xin-Rui,CAO Dong-Li,JIANG Zhi-Sheng,LIU Lu-Shan,WANG Zuo,YI Guang-Hui,WEI Dang-Heng,WANG Gui-Xue and TANG Chao-Ke 1118true Caenorhabditis elegans exhibits chaperone-like activities in vitro. It prevents protein aggregation in a certain extent when use insulin, ADH and lysozyme as the substrates, though it is not as efficient as the typical chaperones (such as HSP16.1 in C. elegans). By contrast, the other three sHSP12s (HSP12.2, HSP12.3 and HSP12.6), which have similar molecular masses and primary structure, appear devoid of in vitro chaperone-like activities. In addition, overexpressing HSP12.1 enhances cell thermotolerance of Escherichia coli. The survival rate of the HSP12.1 overexpressed cells is 4-fold higher than the control, yet whether it does the same function in C. elegans is still unknown. Results indicate that C-terminal region is not necessary for the chaperone-like activity of sHSPs, for HSP12.1 terminates a short C-terminal tail. N-terminal domain may play a relatively important role in the exhibition of chaperone-like activities, while α-crystalline domain may also involve in this function.]]> 2007/5/16 0:00:00 QIN Yan,WANG Hui and CHANG Zeng-Yi 1117true 2007/5/29 0:00:00 TAN Xiao-Jun,ZHANG Yong-Xin,QIAN Min-Ping,ZHANG You-Yi and DENG Ming-Hua 1116true E.coli. BL21(DE3)plysS, which was induced with IPTG (0.4 mmol/L) to express TAT PTD-Ngb fusion protein. Ni-NTA resin was used to purify the product, which was identified by SDS-PAGE and Western blot subsequently. After being purified and desalted, the biological activity of TAT PTD-Ngb was deteced in primary cultured cortical neurons. The results showed that TAT PTD-Ngb could transduced into cortical neurons, increase cell viability under hypoxia and attenuate apoptosis induced by hypoxia. The present study provides a clue for the research of neuroglobin and seems to provide a protein therapy strategy for CNS diseases,especially in cerebrovascular diseases and neurodegenerative diseases.]]> 2007/4/4 0:00:00 ZHOU Guo-Yu,ZHOU Sheng-Nian,LOU Zhi-Yin,ZHU Can-Sheng and HU Xue-Qiang 1115true 2007/5/16 0:00:00 SU Xiao-Zhou,YU Feng,ZHAO Yun-Hui,WU Xiao-Mei and CHEN Jia 1114true Aeromonas hydrophila strain AH-1 by constructing ahyR and ahyI mutant revealed that they are under quorum-sensing control. This observation was supported by the down-regulation of the TTSS genes in the presence of lacZ-TTSS gene promoter and the corresponding advanced secretion of AexT in ahyI mutant.]]> 2007/5/29 0:00:00 ZHOU Wen-Guang,LIU Guang-Feng and ZHOU Shi-Ning 1113true Xenopus oocyte, PAK2-NT(PAK2-N-terminal,PAK2-NT) and PAK2-NTm (PAK2-N-terminal mutation) mRNA were microinjected into Xenopus oocyte respectively. Under fluorescent microscopy germinal vesicle breakdown was observed during cytokinesis. To further observe the relationship of oocyte cytokinesis, polar body formation and Cdc42 activity, confocal microscopy with time-lapse was employed . As a result, occurrences of germinal vesicle breakdown in oocytes were similar to those oocytes injected with PAK2-NT mRNA or injected with PAK2-NTm mRNA,but no cytokinesis and polar body formation were observed in oocytes injected with PAK2-NT mRNA or PAK2-NTm mRNA. These results indicated that PAK2 involved in Xenopus oocytes cytokinesis and polar body formation independent of Cdc42 activity.]]> 2007/5/21 0:00:00 CHENG Da-Ye,LIANG Bin and LI Feng 1112true Arg and pchp53Pro were constructed by inserting expression cassette CMVe+hTERTp+EGFP, CMVe+hTERTp+p53Arg and CMVe+hTERTp+p53Pro into pHrn respectively. 24 h after cell transfection by lipofectamine 2000, GFP expression pattern was analyzed through fluorescence microscope and flow cytometry; RT-PCR and Western blot were taken to study the p53 expression pattern. The cell apoptosis by Hoechst 33258 and Annexin V-FITC/PI staining was also studied. Results show that the expression of GFP and p53 protein in Bel-7402 were detected, but apparent cell apoptosis could not be found. The recombinant p53 mediated by human-derived vector could express in Bel-7402, but no significant tumor suppression effect was detected, which might result from the down regulation effect of the wild type p53 on hTERT promoter.]]> 2007/4/16 0:00:00 XUE Zhi-Gang,LI Jian,YIN Biao,ZHANG Ya-Kun,LIU Xiong-Hao,PAN Qian,LONG Zhi-Gao,DAI He-Ping,XIA Kun,WU Ling-Qian,LIANG De-Sheng and XIA Jia-Hui 1111true OR for DD/ID versus Ⅱ for hypertension among men was significant (OR 1.57; 95% CI, 1.09～2.27; P = 0.016) in dominant genetic model. In combination analysis stratified with respect to gender, slightly significant ORs were found after adjustment in males: OR for TT vs CC, 0.11; 95% CI, 0.01～0.99; P = 0.049 within ACE DD genotype; OR for DD/ID vs Ⅱ, 1.52; 95% CI, 1.01～2.29; P = 0.047 within GNB3 CC+CT genotype. The results suggest that ACE, or a nearby gene, is a male-specific susceptible gene for hypertension, and that there may exist epistatic gene-gene interaction between ACE D allele and GNB3 825C allele.]]> 2007/4/6 0:00:00 HUANG Wen-Tao,YU Hong-Jiang,LU Xiang-Feng,ZHAO Wei-Yan,WANG Yue-Lan,GU Dong-Feng and Chen Run-Sheng 1110true Homo sapien umbilical vein for 24 h. The level of SAM and SAH was detected by HPLC. The activity of SAH Hydrolase was detected by real-time quantitative reverse transcription-PCR and Western blotting analysis. The level and patterns of DNA methylation were measured by endogenous C-5 DNA methyltransferase(C-5 MT-ase) activity and capacity of genomic DNA to accept methy1 groups and methylation-dependent restriction analysis. The results indicated that an increased Hcy concentration induced elevated SAH, declined SAM and the ratio of SAM/SAH, reduced expression of SAH Hydrolase, but increased activity of C-5MT-ase. The methylation status of gDNA analyzed by methyl-accepting capacity of gDNA uncovered a demethylation process in gDNA, or homocysteine-caused hypomethylation in gDNA. With different methylation-dependent restriction endonucleases, the aberrant demethylation was found to prefer C↓CGG sequences to CpG islands. The impacts of different dosage of Hcy showed that the varied detrimental effects of Hcy could be attributed to different concentrations via different mechanisms. In mild and moderate hyperhomocysteinemia, the Hcy may primarily influence the epigenetic regulation of gene expression through the interference of transferring methyl-group metabolism, while in more higher Hcy concentration, the notorious impacts may be more directly caused via oxidative stress, apoptosis, inflammation etc.]]> 2007/4/23 0:00:00 JIANG Yi-Deng,ZHANG Jian-Zhong,HUANG Ying,SU Juan,ZHANG Jing-Ge,WANG Li-Zhen,HAN Xiao-Qun and WANG Shu-Ren 1109true in vitro, but inhibit the neointima formation in rat carotid artery after balloon injury. These properties suggest that hCREG might have played an important role in antagonizing restenosis of vessel by inhibiting phenotypic modulation of VSMCs. In order to further elucidate the biological functions of hCREG in VSMCs, the upstream molecule mechanisms in regulating its expression were analyzed. At first, bioinformatics was used to predict the hCREG promoter and the binding sites of transcriptional factors. According to bioinformatic results, the pEGFP-hCREG-P945 reporter gene vector was constructed successfully which contained upstream 945 bp of hCREG where two binding sites of E2F1 were determined. Subsequently, the hCREG promoter activity was identified directly by detecting the expression of reporter protein-GFP with fluorescence microscope and Western blot analysis when the vector was transfected into human VSMCs-HITASY. Secondly, both the expression of hCREG and smooth muscle α-actin(SM α-actin) was detected to increase in differentiation HITASY cells cultured for 72 h with serum deprivation，in which the expression of E2F1 was reduced significantly. Inversely, the increase of E2F1 expression was detected in dedifferentiation cells cultured with 10% FBS medium accompanied with the reduction of hCREG and SM α-actin. It is suggested that E2F1 maybe inhibit the expression of hCREG by binding to the hCREG promoter. Furthermore, the E2F1 oligodeoxynucleotide (ODN) and mismatch E2F1 ODN were designed and used to block the binding of E2F1 to hCREG promoter by transcriptional factor “decoy strategy”. Western blot analysis showed that expression of hCREG, GFP and SM α-actin was increased obviously when the E2F1 ODN was transfected in dedifferentiation HITASY cells. The result identified that transcription factor E2F1 inhibited the expression of hCREG and promote dedifferentiation of VSMCs by binding to the sites of hCREG promoter. It can be concluded that E2F1, as a transcriptional regulation factor of hCREG, can repress the expression of hCREG and involve in a pivotal role in the process of VSMCs phenotypic modulation.]]> 2007/4/12 0:00:00 Han Ya-Ling,Zhao Xin,YAN Cheng-Hui,KANG Jian,ZHANG Xiao-Lin,DENG Jie,XU Hong-Mei and LIU Hai-Wei 1108true Brassica campestris L. ssp. chinenesis Makino, syn, B. rapa L. ssp. chinenesis) ‘Bpol97-05A’. RT-PCR showed that this fragment was specifically expressed in male sterile material. Sequencing and BLAST search in GenBank database indicated that P1708 had 100% homolog with chloroplast ndhJ-trnF gene region except a 54 bp insertion. Gene specific primer pairs were synthesized according to ndhJ-trnF gene region and two fragments about 1 900 bp were amplified respectively using genomic DNA templates of Polima cabbage and male fertile oilseed rape. The sequencing results showed that the gene region ndhJ-trnF of Polima cabbage contained two 54 bp repeats and some variation sites. The repeat part shared the same sequence as trnF gene except three bases at 5′ ends. For the insertion of 108 bp sequence, a new open reading frame was created.]]> 2007/4/10 0:00:00 JIANG Ming and CAO Jia-Shu 1107true Garcinia hurburyi tree. Here, the cell growth inhibition of TH2 on human hepatocellular carcinoma cell line (Bel-7402) was evaluated in vitro using SRB assay. The treatment of 10 μmol/L TH2 reduced the surviving fraction from 86% (12 h) to 17.2% (48 h). To assess whether TH2 induce apoptosis, the appearance of sub-G1 peak, a specific fraction for apoptosis was detected by flow cytometry analysis. Progressive increase in the percentage of apoptotic population was observed in a dose-and time-dependent manner. Furthermore, a cleavage of poly (ADP-ribose) polymerase (PARP), a marker of early apoptosis, was observed clearly when the cells exposed to 10 μmol/L of TH2 for 24 h by immunoblotting analysis. In vitro activities of 20 S proteasome purified from human erythrocytes on fluorogenic peptide substrates revealed that TH2 inhibited the trypsin-like, chymotrypsin-like and peptidylglutamyl peptide hydrolyzing activities in dose-dependent manner. Moreover, the turnover of tumor suppressor p53, a sign of deregulation of cell cycle progression and apoptosis induction by classical proteasome inhibitors, was disrupted in Bel-7402 cells. All these data indicate that TH2 had inhibitory effect on the proliferation of Bel-7402 cells and induction of apoptosis, which might be related to its inhibition of proteasome.]]> 2007/4/17 0:00:00 XU Bo,XING Cheng,LI Min,GUO Wei and CUI Jing-Rong 1106true GhPRPs have been carried out by quantitative realtime RT-PCR. The results showed that GhPRP3 and GhPRP5 were preferentially expressed in 10 dpa fiber, little transcripts was detected in other tissues examined. GhPRPL highly expressed in cotyledons, whereas smaller or negligible amounts of its transcripts were detected in other tissues. The remaining two genes, GhPRP4 and GhPRP6, were expressed in all the tissues analysed, but their transcript level is different. GhPRP4 mRNA is most abundant in hypocotyls, and then in anther, while GhPRP6 expressed highly in fiber, and then in 10 dpa ovule. Furthermore, the results showed that the fiber-specific GhPRP3 and GhPRP5 were also developmentally regulated, suggesting that the genes may play important roles during cotton fiber development.]]> 2007/4/17 0:00:00 XU Wen-Liang,HUANG Geng-Qing,WANG Xiu-Lan,WANG Hong and LI Xue-Bao 1105true in vivo gene delivery, the cationic polymers are usually further modified by hydrophilic polymer grafting or ligand conjugation, which have been shown to increase the vector stability, gene delivery efficiency and specificity greatly. Some previous research had shown that modified hydrophilic polymer may partly shield the targeting ligand and result in poor delivery specificity. Developing a method to evaluate the influence of PEG modification on targeting delivery is particularly critical to cationic polymer design and gene therapy development. One of most commonly used cationic polymer polylysine (PLL) was chosen as a model. Targeting ligand epidermal growth factor(EGF)was conjugated with PLL to form PLL-EGF. Then hydrophilic polymer polyethylene glycol (PEG) with molecular mass 7 000 and 20 000 were used to modify PLL-EGF respectively to generate PEG7000-g-PLL-EGF and PEG20000-g-PLL-EGF. In BIAcore experiments, epidermal growth factor receptor (EGFR) was conjugated onto BIAcore chip and various PEG modified PLL-EGF solutions were flowed over the chip. By observing the change of RU value, the specific interaction of EGF to EGFR was compared. Compared with PLL-EGF, PEG modified PLL-EGF showed lower association rate and higher disassociation rate to EGFR. Furthermore, compared to PEG7000 modified PLL-EGF, PEG20000 modified PLL-EGF got lower association rate and higher disassociation rate to EGFR. The Scatchard analysis results showed that the interactions between EGFR and PLL-EGF or PEG-PLL-EGF are non-linear. It can be concluded that PEG modification indeed reduced the association rate and enhanced the dissociation rate of EGF to EGFR. The length of PEG chain was also a key factor to influence interaction between ligand and receptor. The results showed that it was critical important to evaluate the influence of PEG modification on delivery specificities. The BIAcore method developed in this paper can successfully evaluate the influence, which would be important for cationic polymer design and its application as potential non-viral gene delivery vectors.]]> 2007/4/16 0:00:00 GUO Yan,GU Jian-Ren and XU Yu-Hong 1104true in vivo were performed and the potential role of AAV2 in β-thalassemia gene therapy was analyzed. The rAAV2 containing a normal human β-globin gene (rAAV2-β-globin) without helper virus contamination were produced. The viral titer, purity and the ability of mediating expression of β-globin gene were detected in vitro. Then, human early fetal liver hematopoietic cells were isolated and were further transducted with the rAAV2-β-globin, followed by transplantation into sublethally irradiated BALB/C nude mice to analyze the β-globin gene expression. The results showed that the high titer and purity of rAAV2-β-globin had the ability of mediating β-globin gene expression in vitro. In 8 recipient BALB/C nude mice, the β-globin gene expression were detected in the 2 mice marrow by RT-PCR. The results suggested that rAAV2 could transduce human fetal liver hematopoietic cells and mediate the β-globin gene expression in BALB/C nude mice, meanwhile the expression level of the gene was still rather low. It is necessary to perform further research on AAV2 biology before applying in β-thalassemia gene therapy.]]> 2007/4/20 0:00:00 SUN Xiao-Juan,TIAN Jing and TAN Meng-Qun 1103true 2007/4/10 0:00:00 Hu Shu-Hong,YANG Yan,ZHANG Jian-Hua,Zhang Mu-Xun and GONG Cheng-Xin 1102true in vitro and in vivo. The recombinant pcDNA3.1(+)/STGC3 plasmid was constructed and transfected into CNE2 cell line with lipofectamine 2000. CNE2 cell clones with stable STGC3 expression were obtained through G418 selection and identified by RT-PCR and Western blotting methods. The effect of β-estradiol on growth rate of pcDNA3.1(+)/ STGC3/CNE2 cell line was examined by cytometry. The pcDNA3.1(+)/STGC3/CNE2 cell line was inoculated subcutaneously in nude mice. Tumorigenicity diversity was analyzed in female and male nude mice. STGC3 expression was detected in tumor xenografts in nude mice by RT-PCR, immunohistochemistry and Western blotting methods. Cell morphologic changes were observed by microscope. Flow cytometry was used to analyze cell cycles. The results indicated that pcDNA3.1(+)/STGC3/CNE2 cell growth rate decreased after treated with β-estradiol in vitro. There were significant differences in tumor size and mass between pcDNA3.1(+)/ STGC3/CNE2 and control cases (P < 0.05). Furthermore, tumor size and mass had significant differences between male and female nude mice in the STGC3 transfection cases and tumor growth rate of pcDNA3.1(+)/ STGC3/CNE2 in the female nude mice was the lowest in all cases (P < 0.05). No significant difference was found between male and female nude mice in other control cases (P > 0.05). There were more cells blocked in G0/G1 phase in tumor xenografts of pcDNA3.1(+)/STGC3/CNE2 cell line of the female nude mice case than that in other cases. Data above indicated that estrogen might enhance the effect of STGC3 to inhibit CNE2 cell proliferation in vitro and in vivo.]]> 2007/4/6 0:00:00 HU Bo,QIU Qing-Chao,HE Xiu-Sheng,LUO Qiao,TANG Guo-Hua,LONG Zhi-Feng and LIAO Yin-Hua 1101true in vitro. To examine the effect of STGC3 on the tumorigenicity of CNE2 cell line and explore its mechanism in nude mice. The Tet/pTRE/CNE2-STGC3 cell line was planted under the front leg skin of nude mice and induced by doxycycline (Dox). The mRNA and protein level of STGC3 in transplanted tumor tissues were detected with RT-PCR and Western Blotting. The apoptosis ratio of the tumor cell was analyzed with flow cytometry. STGC3, Bcl-2 and Bax proteins were examined by immunohistochemistry method. The results indicated that high level of STGC3 expression can inhibit tumorigenicity of CNE2 cell line in nude mice. Tumor grew slowly, later and smaller. Cell apoptotic percentage increased. Bcl-2 protein expression was down-regulated and Bax protein expression was up-regulated in Tet/pTRE/CNE2-STGC3 cell line (P < 0.01). The data indicated that STGC3 had a role as a tumor suppressor gene in vivo, which was in line with the results in vitro.]]> 2007/3/27 0:00:00 QIU Qing-Chao,HU Bo,HE Xiu-Sheng,LUO Qiao,LONG Zhi-Feng,TANG Guo-Hua and LIAO Yin-Hua 1100true 2007/3/21 0:00:00 Yang Yi-Xin,YANG Yun-Bo,LI Xiao-Ling,PENG Cong,GUO Qin,SHEN Shou-Rong and LI Gui-Yuan 1099true in vitro and in vivo. Compared with wild type MUC1, the mutants depressed soft agar colony formation and showed abrogated tumorigenicity in nude mice. These findings implicate that CQCRRK motif mediate the formation of MUC1 protein complex. As a result of this research, disruption of MUC1-C-terminal subunit-associated dimerization by mutation of CQC→AQA might represent a novel therapeutic approach for tumor.]]> 2007/3/20 0:00:00 WEI Ling,YI Yan-Ping,LIU Xuan,DONALD KUFE and CAO Cheng 1098true 0TFPI, m1TFPI, and m2TFPI, were generated by introducing mutations in the DNA sequences coding for the C-terminal amino acid sequences. The clearance rate of the experimental group (three rTFPI mutants) and the control group (wild-type TFPI, namely mTFPI) in rats was investigated. The inhibitory function on FXa and the heparion binding capacity of the TFPI mutants were also studied. It was shown that at 10 min after intravenous injection, (59.06 ± 13.54) % of the injected mTFPI remained in the circulation, while (71.14 ± 13.04) % of the m0TFPI, (77.99 ± 2.53) % of the m1TFPI, and (82.95 ±11.36) % of the m2TFPI were still in the circulation. The relative half-life of m0TFPI, m1TFPI and m2TFPI were prolonged 0.5, 0.9, and more than 1 times respectively in comparison with that of mTFPI, while their FXa inhibiting activity and heparin-binding capacity were little changed.]]> 2007/3/2 0:00:00 YU Bo,LUO Shi-Ping,SONG Li-Ping,LENG Xi-Gang,ZHAO Ming-Hui and NIU Hui-Sheng 1097true 2007/3/27 0:00:00 CUI Jun,FU Han-Jiang,FENG Jun-Jun,ZHU Jie,TIE Yi,XING Rui-Yun,WANG Cheng-Fu and ZHENG Xiao-Fei 1096true 2007/3/21 0:00:00 ZHENG Bin,WEN Jin-Kun,HAN Mei and SHI Jian-Hong 1095true 2007/3/9 0:00:00 FENG Zhou-Yan,GUANG Lei,ZHENG Xiao-Jing,WANG Jing and LI Shu-Hui 1094true 6 U/mg level after expression and purification and the bioactivity of A is higher than that of B. The results proved primarily that the design was reasonable and provided premise and basement for CNTF's development and application.]]> 2007/3/20 0:00:00 BI Hua,YUE Jun-Jie,YUAN Li-Yong,RAO Chun-Ming,CHEN Hui-Peng and WANG Jun-Zhi 1093true 2007/3/27 0:00:00 HE Cheng-Qiang,LI Cong-Cong,WU Yan-Hong,AN Li-Guo and LI Yun-Long 1092true E. coli, thereby facilitating the analysis of viral gene functions in the context of genome. This new technique has provided a useful tool for the study of pathogenesis mechanism of EBV, especially for that of cancer-associated.]]> 2007/3/19 0:00:00 TANG Yun-Lian,LU Jian-Hong,WU Ming-Hua,HUANG Chen,CAO Li,PENG Shu-Ping,ZHOU Yan-Hong,LI Xiao-Ling,ZHOU Ming,TANG Ke and LI Gui-Yuan 1091true 304 cells), total RNAs were extracted with TRIzol from adipose tissues of human, then the 1 102 bp fragment of SelS was amplified by RT-PCR. After the expected 1 102 bp PCR fragment was purified, SelS was cloned into pMD18-T vector. Purification, RT-PCR, restriction endonuclease analysis and DNA sequencing were performed to confirm the results. pMD18-SelS was incubated in presence of XhoⅠ/ClaⅠ and then ligated into pLNCX2 vector corresponding restriction sites. RT-PCR and DNA sequencing were performed to confirm a 1 102 bp SelS gene fragment was successfully inserted into pLNCX2. The ECV304 cells were stably infected with pLNCX2-SelS or pLNCX2 alone and positive clone was obtained by G418 selection. After transient transfection, ECV304 cells expressed pLNCX2 was verified by neor gene detection and the expression of recombinant SelS in the ECV304 cells was 1.76-fold higher than the endogenous level by RT-PCR. After the cultured ECV304 cells was treated with hydrogen peroxide (H2O2, 100, 200, 300, 400 μmol/L) for 24 h, cell viability was measured by MTT assay and the intracellular maleic dialdehyde(MDA)was detected by TBA method. The results showed that ECV304 cells were injured by H2O2, overexpressing SelS increased the cell viability apparently and inhibited the production of MDA induced by H2O2. So overexpressing SelS protects ECV304 cells from injuring by H2O2, and SelS may have the antioxidant protection effective on endothelial cells.]]> 2007/3/27 0:00:00 DU Jian-Ling,AN Li-Jia,SUN Chang-Kai,MEN Li-Li,ZHANG Xiu-Juan and LI Chang-Chen 1090true 10-dB value and symmetry index >0 of the FTC, increasing of the proportion of CF shift toward higher frequency side (HFS) of neuron's FTC is much higher. There is significant correlation between the time course and the age of rat, the older is the age, the longer is the shift time (P < 0.05) and the recovery time (P < 0.05) of neurons. The results suggest that plasticity of characteristic frequency in rat auditory cortical neurons is correlated with the postnatal age. The findings provided important data to the study of the mechanisms for the developmental plasticity of central nervous system.]]> 2007/3/27 0:00:00 TAN Jiang-Xiu,WANG Fang,YANG Wen-Wei,ZHANG Ji-Ping and SUN Xin-De 1089true 412) quickens, this phenomenon is not seen when metal cations are added to blue omembrane solution(the rate of decay of photcycle intermediate slowers). At the same time , the effect of pH and temperature on the interaction between hydrazine and blue membrane was investigated. When hydrazine was added to blue membrane solution, the sensitivity of the reaction is pH and temperature dependent. Over the pH range(2～4.8), the sensitivity of the reaction lowers with the increase of the acidity. Over the temperature range(20～40℃), the sensitivity of the reaction lowers with the increase of the temperature.]]> 2007/3/2 0:00:00 su tao,zhang yue,hu jian-qi,luo zhi-yong and hu kun-sheng 1088true in vivo biological function of Opg and the mechanism of osteoporosis due to deficiency of Opg, Opg knockout mice have been generated through homologous recombination. Opg－/－ mice exhibit a sharply decrease in bone density and strength as expected. The number of osteoclasts in Opg－/－ mice significantly increases. Morphologically, osteoclasts appear more cuboidal in shape in Opg－/－ mice than those of wt mice, suggesting that active osteoclastogenesis occurs in the absence of Opg. In consistent with this finding, an increase of osteoblast activity was also observed with accelerated mineral accumulation rate by histomorphometric measurement and elevated serum alkaline phosphatase activity (ALP) in Opg－/－ mice. Interestingly, more than 50% of 2-month-old Opg－/－ mice manifest medial calcification of aorta with comparable serum concentrations of calcium and phosphorus to wt mice. In conclusion, Opg－/－ mice have a high-bone-turnover type osteoporosis. The aortic calcification in Opg－/－ mice is not due to abnormality of calcium and phosphorus metabolism. The mechanism underlying aortic calcification in Opg－/－ mice needs to be further investigated.]]> 2007/2/6 0:00:00 XU Yong,YANG Hua,QIAO Jian-Ou,Li Xi-Hua,YAN Lan-Zhen,WANG Long,XU Guo-Jiang,FEI Jian,Fu Ji-Liang and WANG Zhu-Gang 1087true 2007/2/8 0:00:00 XIA Hong-Fei,SUN Jing,YANG Ying and PENG Jing-Pian 1086true 1C subunits in NSCs, but no Cav1.3α1D subunits. Moreover, L-type calcium channel currents were recorded in those cells by using whole-cell patch clamp techniques. It was found that activation of L-type calcium channels promotes proliferation and differentiation to neuronal type of NSCs. The results indicated that rat embryonic hippocampal NSCs express functional L-type calcium channels, L-type calcium channels modulate proliferation and differentiation of NSCs.]]> 2007/2/8 0:00:00 FENG Jin-Li,HU De-Hui and GAO Tian-Ming 1085true Schistosoma japonicum. To observe the in vitro expression and subcellular localization of recombinant eukaryotic expression plasmid pEGFP-C1/ SJCHGC in COS-7 cells and analyze the antigenicity of the products expressed. 9 pairing-related proteins were identified and further validated by semi-quantitative RT-PCR analysis. Most of the candidate proteins have been evident to be somehow associated with various aspects of S. japonicum such as cell growth, development, motility, reproduction, nutrition and signal transmission. The SJCHGC gene was expressed in the transfected COS-7 cells. The expression and subcellular localization of the fusion protein were analyzed by fluorescence microscopy. The proteins expressed show the antigenicity of Schistosoma japonicum. These results provide the basis for elucidating the Schistosome mechanism of development and reproduction, and may permit the identification of protein candidates for the development of novel vaccine or new targets for drug development against schistosomiasis.]]> 2007/2/9 0:00:00 DAI Gan,WANG Shi-ping,YU Jun-Long,XU Shao-Rui,PENG Xian-Chu,LIU Xue-Qin,ZHOU Song-Hua and LIU Fen 1084true Saccharomyces cerevisiae. GRE3 gene encoding an NADPH-dependent aldose reductase was chosen as an example. A recombinant plasmid psiLENT-GRE3 was constructed based on the pESC-LEU backbone and used to transform S. cerevisiae YPH499. The down regulation of GRE3 gene expression by inducing 1 kb RNA duplex and a 136 bp loop was investigated using reverse transcription - PCR. The results showed that double-stranded RNA mediated gene silencing could be used as a functional tool to decrease the expression level of a specific gene in S. cerevisiae, which would contribute to the understanding of RNA interference in budding yeast.]]> 2007/2/8 0:00:00 LI Yan,YAN Ming,WANG Kui,XU Lin,XU Wei and OUYANG Ping-Kai 1083true Ganoderma lucidum (Se-GL-P), the primary biochemical characters of Se-GL-P were characterized. Molecular mass was determined by two methods of sodium dodecyl sulfate polyacrylamide gel electrophoresis and size exclusion chromatogram. Amino acids composition, isoelectric point, peptide mass fingerprint, and the sequence of N terminal amino acids were determined by high performance liquid chromatography, capillary isoelectric focusing method, in-gel digestion, and Edman digestion method respectively. Results showed that, the Mr of Se-GL-P was 36 600, pI = 4.01, and the sequence of N-terminal amino acids was DINGGGATLPQKLYLTPDVL. A conclusion was drawn that Se-GL-P was a kind of new selenium-containing protein, and it was one member of DING protein family.]]> 2007/2/13 0:00:00 du ming,hu xiao song,wang cong and zhao guang hua 1082true E. coli SD108(topA+ gyrB225). Now, this structure is also found in E. coli DM800(topA－ gyrB225). The result indicates that the formation of supercondensed DNA is related with decrease of the activity of gyrase in vivo. Topoisomerase Ⅳ was proved to relax the supercondensed DNA completely in vitro, which suggested that the supercondensed DNA and the supercoiled DNA could transform to each other in cells. The supercondensed DNA samples were analyzed by atomic force microscopy and compared to supercoiled DNA. The results showed that the length of supercondensed DNA decreased about 30% and the width and height of double-strand increased about 60%, which indicates that the structure of double-strand of supercondensed DNA is much more similar to A-DNA than B-DNA. The results also showed that chloroquine intercalation did not change the supercoiling level of supercondensed DNA, but made it knot and compact.]]> 2007/2/6 0:00:00 ZHANG Zhen-Feng,YU Jia,CAO Kou and HUANG Xi-Tai 1081true 2007/2/12 0:00:00 YAO Cui-Ping,ZHANG Zhen-Xi and Yao Bao-li 1080true 2007/2/9 0:00:00 JIA Jian-An,ZHOU Bo,JIANG Shao-Hua,CHEN Qiu-Li,ZHAO Ping,PAN Xin,WEN Zong-Mei,DENG Song-Hua,LU Hong-Zhou and PAN Wei 1079true 2 in 24 h, and step-wise increased shear stress from 0.5 to 1.0 and then to 1.5 N/m2 and each stress for 8 h, and stepwise decreased shear stress from 1.5 to 1.0 and then to 0.5 N/m2 and each stress for 8 h were applied. The expression of OPG and ODF mRNA was detected by RT-PCR, and synthesization of their protein was detected by Western blot. The results showed that the shear stress of 1.0 and 1.5 N/m2 induced more significant changes in OPG mRNA and ODF mRNA than 0.5 N/m2 and there was no significant difference between 1.0 and 1.5 N/m2. No marked differences were observed between the effects of step-wise increased shear stress and 1.5 N/m2 after 24 h, and the same was between the effects of step-wise decreased shear stress and 0.5 N/m2 after 24 h. The shear stress of 1.0 N/m2 increased OPG expression and inhibited ODF expression in 24 h both in mRNA and protein level. In all, the ratio of OPG and ODF was increased by shear stress, which reminds that bone resorption was inhibited by the fluid flow, and shear stimuli regulated the new balance of bone resorption and bone formation throught OPG/ODF route.]]> 2007/2/12 0:00:00 ZHANG Bing-Bing,PAN Jun,WANG Yuan-Liang,XIAN Cheng-Yu,XIANG Yan,XIN Juan,LI Yong-Gang and WANG Xi-Yun 1078true via TLR4 signaling pathways and activated NFκB signaling pathway, which can further lead to nasopharnyx inflammation and other nasopharnyx disorders.]]> 2007/1/16 0:00:00 YANG Yi-Xin,YANG Yun-Bo,ZHOU Ming,ZHOU Hou-De,XIANG Bo,PENG Shu-Ping,LI Xiao-Ling,SHEN Shou-Rong and LI Gui-Yuan 1077true in vitro. The RCCS can simulate microgravity effects with low shear stress and well exchanging for gas. Then the growth characteristics and some mechanism of the cells in RCCS were detected by scanning electron microscopy (SEM), transmission electron microscopy (TEM), RT-PCR and flow cytometry (FCM). The results indicate that the cells grew well with polyhedron morphology and lots of microvilli, mitochondria and tight junctions in this system, which means that this system is useful for cells to form 3D structure to mimic cell status in vivo. The expression of some cell adhesion molecules (CAMs) were changed markedly, which are closely associated with cancer invasion and metastasis. The characters of increased expression of integrin β1(CD29), CD44, intercellular adhesion molecule-1(CD54) and depressed expression of E-cadherin presumably show that the HepG2 cells cultured in RCCS could recur some characters of primary liver cancer in vivo, the capacity of invasion and metastasis. It is necessary for acquiring perfect and external results to select an appropriate research model for studying in vitro. This 3D culture in vitro under simulated microgravity can provide a useful and reasonable model for oncology, anticancer drugs research and other research.]]> 2007/1/16 0:00:00 XU Da-Yong,WANG Yun and FENG Mei-Fu 1076true de novo miRNA precursor prediction method was proposed. In constructing the recognition model, both primary sequence and secondary structure were combined into an input sequence through encoding, and the input space was mapped into a feature space via k-gram method. After applying feature selection, those selected features was used to construct SVM-based models for the recognition of miRNA precursors. In the mean time, the method was compared with the HMM learning method. Experimental results show that the method outperforms HMM. The reason is that microRNAs are so short that it is not easy for HMM model to capture the signals for differentiating the genuine microRNAs from those pseudo-microRNA genes. From features selected, it was found that they are mostly come from the primary and secondary structure of microRNAs. This phenomenon may tell us to put more efforts in the microRNAs themselves in designing computational method before we fully understand the transcription mechanism of microRNA biologically.]]> 2007/1/18 0:00:00 YANG Liang-Huai,Lü Pi-Ming,CHEN Li Jun and DENG Ming Hua 1075true 2007/1/15 0:00:00 YANG Xiao-Ya,BAI Yun,WANG Yan-Yan,XU Xue-Qing and CHEN Xue-Dan 1074true 2007/1/17 0:00:00 Li yanling,LI Xiang-Ning,ZHOU Wei,ZENG Shao-Qun,LIU Man and LUO Qing-Ming 1073true smu.776 encodes a possible S-adenosylmethionine-dependent methyltransferase of 385 residues in Streptococcus mutans, a primary pathogen for human dental caries. The DNA fragment of smu.776 was cloned into pET28a and expressed in good amount from the E. coli strain BL21 (DE3). Smu.776 protein was purified to homogeneity in a two-step procedure of Ni2+ chelating and size exclusion chromatography. Crystals were obtained by hanging-drop vapor diffusion method and diffracted to 2.0 Å resolution. The crystal belongs to orthorhombic space group C2 with cell dimension of a=168.47 Å, b= 50.66 Å, c=53.96 Å, β=104.22°. The asymmetric unit is expected to contain one molecule with solvent content of 51.3%.]]> 2007/1/16 0:00:00 WANG Kai-Tuo,LI Lan-Fen,ZHANG Xiao-Yan,LIANG Yu-He,WEI Shi-Cheng and SU Xiao-Dong 1072true Staphylococcus epidermidis is the common pathogen of nosocomial infection. SarA (staphylococcal accessory regulator A) is a global factor regulating expression of most virulence genes in S. epidermidis. It is reported that SarA regulates the transcriptions of lipA encoding lipase, zinC encoding membrane- associated zinc metalloprotease and atlE encoding autolysin. SarA exhibiting an up-regulation on lipA and zinC, and down-regulation on atlE were demonstrated by reverse-transcription PCR and reporter gene lacZ analysis respectively. The results demonstrate that SarA regulated directly the expressions of lipA, zinC and atlE by protein-DNA binding to the promoter region of AT-rich sequence. The consensus sequence of binding sites in atlE, lipA and zinC promoter regions of S.epidermidis RP62A was forecasted by using program Omiga 2.0. The results suggest that SarA could be treated as the target in the prevention of Staphylococcus epidermidis infection.]]> 2007/1/16 0:00:00 WANG Hai-Jiao,FAN Chang-Sheng,XIN Ji-Di,TAO Ju-Hong,YUAN Xing-Hui,GAO Shan-E and LIANG Guo-Xin 1071true 32P of [γ-32P]ATP into the substrate. The mRNA expression of CK2α, α' and β subunits were detected by multiplex RT-PCR. CK2 kinetic analysis was carried out by using the Lineweaver-Burk plot. Digitoflavone was shown to inhibit strongly the activity of recombinant human CK2 holoenzyme in vitro (IC50=0.86 μmol/L) and endogenous CK2 in HL-60 cells, which was more effective than the positive control 4Br-2-azabenzimidazole (TBB). After being treated for 2 h with digitoflavone, the mRNA expression of CK2α and α' subunit decreased, while β subunit was not changed fundamentally. Kinetic studies of digitoflavone on recombinant human CK2 showed that digitoflavone acted as an inhibitor of competitive with ATP and mixed types with casein. These results indicating that digitoflavone is a potent endogenous inhibitor of protein kinase CK2.]]> 2007/1/17 0:00:00 LIN Xiao-Cong,LIU Xin-Guang,RUAN Jie,CHEN Xiao-Wen and LIANG Nian-Ci 1070true 2007/1/16 0:00:00 DAI Zhi,LIU Yin-Kun,GUO Kun,SUN Qiang-Ling,LU Wen-Jing and YANG Peng-Yuan 1069true Streptococcus mutans, a primary pathogen for human dental caries. The specific function of Smu_195c is still unknown and there are no conserved domains in it. In order to find out its function, the gene encodes Smu_195c was cloned and expressed in E. coli as N-terminally 6*His tagged recombinant protein. Two crystal forms were obtained by the hanging drop method. Form Ⅰ belongs to space group P6122 or P652 with the unit cell parameters a = b = 62.93 Å, c = 90.63 Å, γ=120° and form Ⅱ belongs to the space group P41212 or P43212 with the unit cell parameters a = b= 57.97 Å, c = 103.51 Å. Crystals from the protein with His-tag belong to form Ⅰ, however, crystals from the protein without His-tag belong to both.]]> 2007/1/16 0:00:00 GAO Zeng-Qiang,HOU Hai-Feng,LI Li-Qin,XU Rui,LIANG Yu-He,LI Lan-Fen,SU Xiao-Dong and DONG Yu-Hui 1068true in vitro by supplying an eligible microenvirnment free of animal feeder layers.]]> 2007/2/6 0:00:00 XI Jia-Fei,WANG Yun-Fang,ZHANG Peng,YUAN Hong-Feng,SHI Shuang-Shuang,YAN Fang,CHEN Lin,BAI Ci-Xian,NAN Xue and PEI Xue-Tao 1067true in vivo. Further phenotype analysis is under the way.]]> 2007/1/16 0:00:00 YANG Zhi-Feng,WANG Long,YANG Sheng-Sheng,KUANG Ying,CHEN Huan,XU Guo-Jiang,CAI Zai-Long,WANG Zhu-Gang and MAO Ji-Fang 1066true Xenopus Paraxial Protocadherin (PAPC), which was initially identified in a screen for genes present in the Spemann organizer of Xenopus embryos, is required for gastrulation, somitogenesis and otic vesicle formation. In order to investigate its function in various developmental events, an antibody was prepared which could specifically recognize Xenopus PAPC. Glutathione S transferase (GST) expression system was used to express the fusion protein GST-PAPC. Rabbits were immunized with GST-PAPC fusion protein and the anti-PAPC polyclonal antibody was obtained. In dilution of 1∶3 000 the antibody recognized a specific band in Western blotting analysis of FL-PAPC transfected HEK 293T cells lysates, which could be specifically blocked by pre-adsorption of prokaryotic expressed GST-PAPC fusion protein. Furthermore, by using immunofluorescence analysis the polyclonal antibody recognized membrane-bound PAPC in FL-PAPC transfected 293T cells and Xenopus animal cap cells. By Western blotting analysis, the endogenous 150 ku PAPC protein was detected in Xenopus embryos using the anti-PAPC antibody. Take together it could be concluded that a polyclonal antibody specifically against Xenopus PAPC was developed.]]> 2007/2/6 0:00:00 HU Rui-Ying,XU Peng,CHEN Yue-Lei,LOU Xin and DING Xiao-Yan 1065true 2007/1/15 0:00:00 LUO Ming-Yong,TIAN Zhi-Gang,XU Zhi,ZHANG Liang,WANG Ying-Xiong and CHENG Jing 1064true P < 0.01). Furthermore, the expression of HSP70 protein was increased remarkably by transfection with a full length HSP70 recombinant plasmid into A549 cells. MTT assay revealed that HSP70 over-expression could block significantly the inhibition of proliferation induced by Act D, there had statistic significance between HSP70 over-expression group and only Act D-treated group (P < 0.01). Furthermore, over-expression of HSP70 could significantly inhibit Act D-mediated apoptosis in A549 cells with a lower percentage of apoptotic cells (4.25±1.48)% in HSP70 transfection group, (12.89±2.03)% in vector transfection group and (14.37±2.56)% in Act D treatment group and disappearance of DNA laddering. These results suggests that increased levels of HSP70 in lung adenocarcinoma tissues decrease the sensitivity of lung adenocarcinoma to Act D, promote proliferation and suppress apoptosis of lung adenocarcinoma cells.]]> 2006/12/14 0:00:00 Liu Xian-Ling,LIU Fu-You,LI Gui-Yuan,HU Chun-Hong and MA Fang 1063true 2007/1/15 0:00:00 LI Zhi-Qing,LIU Li-Xin,LV Feng-Xiang,MA Jing and GAO Guang-Xia 1062true 1～10 mRNA in the human pleural mesothelial cells, culture reproducible model of human pleural mesothelial cells was established in vitro.The value of aquaporin water channels was investigated in pleural fluid dynamics. Mesothelial cells were isolated from pleural effusion fluid of nonmalignant pleural effusion patients and cultured in vitro. The mesothelial cells were identified by morphology and streptomyces protein-peroxidase(S-P) procedure. RT-PCR was used to detect the expression of AQP1～10 mRNA in human pleural mesothelial cells. Establishment of culture reproducible model achieved success of human pleural mesothelial cells in vitro. Immunocytochemistry revealed that cytokeratin and vimentin expressed positive, VⅢ factor associated antigen and CD45 was negtive. Confluent human pleural mesothelial cells appeared multipolar and like cabblestone.Under electron microscopy numerous surface microvilli and abundant endoplasmic reticulum were observed. The cells was identified that it was mesothelial cells. The cells isolated from human pleural effusion fluid have higher purity quotient . RT-PCR studies showed that all of the AQP1～10 mRNA expressed in mesothelial cells. AQP1, AQP9, AQP10 mRNA have abundant expression in mesothelial cells and significant deviation was found between AQP1, AQP9, AQP10 and AQP2, AQP3, AQP7, AQP8. Human pleural mesothelial cells expressed AQP1～10 mRNA, all of them have relation with the function of human pleural mesothelial cells. It was confirmed that mesothelial cells have important contribution in pleural fluid dynamics. Aquaporins (AQP) are involved in rapid and active gating of water across biological membranes.Expression of aquaporins was intervened, new treat method may be found of pleural effusion.]]> 2007/1/15 0:00:00 WANG Hong-Feng,LIU Hong-Bo,QU Wen-Xiu,HE Ping,WANG Zhi-Yun and LI Sheng-Qi 1061true 2007/1/15 0:00:00 ZHENG Jie,ZHU Guo,CHEN Zhu-Chu,LI Ming,CHENG Ai-Lan,RUAN Lin,LIU Ying-Fu,YUAN Wei-Jian,ZHANG Peng-Fei and XIAO Zhi-Qiang 1060true SfiⅠ, and inserted into pComb3XSS. Positive phage-displayed antibody clones were selected on live cell lines and immobilized protein. The purified Fab was verified by SDS-PAGE and Western blot, which showed two bands at about 25 ku and 27 ku at the expected sizes. To analyze the immunological characters of Fab for Met binding, flow cytometry and immunoprecipitation assays were set up and carried out with S114, MKN45 and NIH3T3 cell lines. The results demonstrated Fab could bind native Met specifically on the S114 and MKN45 cell surface. In vitro cytotoxic assay showed that only Hum-ZAP/anti-Met Fab complex could inhibit the Met positive cell growth, it illuminated that anti-Met Fab bound the Met on Met positive cell surface and were internalized. For Met negative cell line NIH3T3, no significant inhibition among the different dosages of Fab and Hum-ZAP. The results showed that anti-Met Fab antibody fragments could recognize Met extracellular domain in native conformation with relatively high affinity. Importantly, these antibody fragments were able to be internalized into Met over-expressed tumor cell lines through binding to the Met receptor, and could be applied as a potential powerful reagents for clinical therapy.]]> 2006/12/13 0:00:00 ZHU Jin,ZHAO Ping,JIAO Yong-Jun,WANG Xin,CAO Bo-Liang,FENG Zhen-Qing and GUAN Xiao-Hong 1059true via the rAAV8 vector injected intraperitoneally into mice at the dose of 5×1010 gc/ mouse. Considerable expression was detected in mouse plasma at two weeks after the administration. Expression peak up to 1 000% level compared to the UCRP occurred between 1～2 months after administration, and then the expression declined gradually but obvious expression persisted at 4 months post administration. APTT test proved the clotting activity of the recombinant hFⅨ in mouse plasma. Immunohistochemical assay showed that the human coagulation factor has been expressed significantly in several organs including liver, kidney, heart and muscles (abdominal and hindleg). These suggest that the rAAV8 virus injected intraperitoneally has been delivered via blood circulation and transduced different cells of multi-organs, and the product of the gene mediated by the rAAV8 vector is biologically active.]]> 2007/1/15 0:00:00 CAO Zuo-Wu,LI Guan-Gui,LIANG Yu-Qiang and HE Dong-Mei 1058true Lactobacillus diolivorans(DSM14421)under the anaerobic condition，respectively. Its putative genes encoding diol dehydratase, a key enzyme in the pathway, were sequenced. However, their reactivating factor (gldG and gldH) was not completely sequenced. Based on several glycerol dehydratase and diol dehydratase reactivating factors sequence alignment, it was amplified from L. diolivorans with degenerated primers. Then it was inserted into expression vector pSE380. A recombinant plasmid pSE-gldGH was constructed and transformed into Escherichia coli BL21. Recombinant gldG and gldH protein were co-purified by metal chelating affinity chromatography and gel filtration from cell-free extracts of E. coli overexpressing the gldGH genes. They existed a putative reactivating factor, with an apparent molecular mass of 325 000. The protein complex consisted of equimolar amounts of the two subunits with Mr of 68 000 (α) and 13 000 (β), encoded by the gldG and gldH genes, respectively. Therefore, its subunit structure was most likely α4β4, different from the common structure α2β2 of the other diol or glycerol dehydratase reactivating factors. In the presence of free adenosylcobalamin, ATP, and Mg2+, the factor reactivated diol dehyadratase from L. diolivorans, which had been inactivated to be enzyme-cyanocobalamin complex.]]> 2007/1/15 0:00:00 MENG Xiao-Lei,Tang Yue,QI Xiang-Hui,WEI Yu-Tuo and HUANG Ri-Bo 1057true E. coli with high efficiency, purification using Glutathione Sepharose 4B and Sephacyl S-200, the recombinant CKLFSF2 C51 protein (typically >95% pure) was obtained. The N-terminal animo acid sequencing of CKLFSF2 C51 was performed and relative molecular mass is 5871.48 measured by ASIMA-CFRTM plus MALDI-TOF. ELISA assay detected the reactivity of the polycolonal antibody and Western blot show that CKLFSF2 was overexpressed in mammalian cells and the specific band was corresponding to the expectation. The recombinant CKLFSF2 C51 protein exhibits significantly chemotactic effect in PC-3 cells, which can be neutralised by anti-CKLFSF2 C51 polyantibody. The anti-CKLFSF2 C51 antibody can also be used for immunohistochemistry. Taken together, the recombinant CKLFSF2 C51 from E.coli has similar bioactivity as that from eukaryocytic expression system. The polycolonal antibodies of recombinant CKLFSF2 C51 can be used in immunohistochemistry, Western blot and to neutralize the chemotactic effect.]]> 2007/1/15 0:00:00 ZHONG Ying-cheng,SHI Shuang,ZHANG Ying-mei,MO Xiao-ning,Liu Da-zheng,SONG Quan-sheng,HAN Wen-ling and WANG Ying 1056true 2007/1/15 0:00:00 Liang Ke,CHEN Zhu-Chu,YI Hong,LI Jian-Ling,ZHANG Peng-Fei,LI Mao-Yu,LI Cui,FENG Xue-Ping,PENG Fang and XIAO Zhi-Qiang 1055true in vitro fertilization (IVF) and parthenogenetic embryos were subjected to immunocytochemistry with antibody to DNA methylation (5'-MeC). In order to research the regulation of oocyte cytoplasm to the relative expression level of imprint genes, the relative expression level of U2afbp-rs gene which is maternally imprint and non-imprint eIF-4C gene in pre-implantation IVF, parthenogenetic and parthenogenetic morula nuclear transplantation embryos by real-time PCR were detected.The results show that neither the genomic DNA of NIH3T3 cloned embryos nor parthenogenetic morula nuclear transplantation embryos were demethylated actively. Although both the relative expression level of U2afbp-rs gene and eIF-4C gene in parthenogenetic morula nuclear transplantation embryos were lower than control parthenogenetic embryos, the expression of the two genes were similar to the control parthenogenesis group. These results suggest that the imprinting gene expression of donor nuclear are regulated by oocyte cytoplasm.]]> 2007/6/11 0:00:00 LI Shao-Hua,YU Jian-Ning,WANG Dan-Qiu,WU Wei,LIN Fei and LIU Hong-Lin 1054true PRSS1) were amplified by PCR and it?蒺s products were analyzed by sequencing, related clinical data were also collected. All the four patients were found mutations in the PRSS1 gene but their clinical feature is absolutely different. Six patients with diabetes mellitus were found in pedigree No. 1, it's members show pancreatitis symptom later, at about 29, the tumor markers (CA19-9, CA72-4) is obviously higher than the patients in pedigree No. 2, two patients with chronic pancreatitis in pedigree No. 2, show symptom earlier without diabetes mellitus, their clinical characterization are different too. The number of CD4+T cell/ CD8+T is very low in Ⅲ8, but Ⅲ7 is normal, and the level of anti-HBs of Ⅲ8 is variable in the course of pancreatitis, but the phenomenon was not found in Ⅲ7. In their PRSS1 gene two guanosine (G) to adenosine (A) mutations were found in PRSS1 exon 3 of pedigree No. 1, one was detected at 336 basyl, the other mutation occurs at 361 basyl. The results of the mutations were Lys →Lys and Ala →Thr. While thymine (T) to adenosine (A) and (guanosine) G→(adenosine) A mutation in PRSS1 exon 3 was detected in the other patient of pedigree No. 2 (Ⅲ8). One was 361 basyl, the other at 415 basyl. While c.415 T→A was not found in the proband of pedigree No. 2 PRSS1 gene (Ⅲ7). All of the mutations were heterozygous mutation, that is to say all of the trypsinogen were wild type and mutant type concomitance, the normal and abnormal pathway of active trypsinogen exist partially. At the same time, the mutations of SPINK1 were not observed. Compared with the documents and registration of NCBI, it can be concluded that PRSS1 gene had many kinds of mutations in hereditary pancreatitis, the heterozygous mutations (c.336 G→A, c.415 T→A) were the novel mutations and related with clinical phenotype. What's more, it's the first time that the multisite heterozygous mutations of PRSS1 gene were reported. The presence of the mutations in four patients with chronic pancreatitis, it's absence in their relatives and the strong evolutionary conservation of the mutation, all indicate that the trypsinogen mutation is associated with hereditary pancreatitis and for the first time raises the question whether a gain or a loss of trypsin function participates in the onset of Chinese pancreatitis.]]> 2007/12/19 0:00:00 LIU Qi-Cai,GAO Feng,ZHUANG Ze-Hao,YANG Bin,LIN Shou-Rong and YIN Qiang 1053true PIC=0.621～0.640) and the genetic diversity of 41 captive Giant Pandas was higher (a population: A=5.48, Ho=0.475, He=0.690; b population: A=5.24, Ho=0.453, He=0.719; captive wild population: A=3.80, Ho=0.514, He=0.725) than that of other 6 endangered species(Ho=0.210～0.390, He=0.150～0.430), but was lower than that of 3 non-endangered species. 41 captive Giant Pandas maintenanced high genetic diversity. However, compared to 7 captive wild Giant Pandas, the genetic diversity of 34 captive Giant Pandas degraded. The date of F-statistics and gene flow (of 25 microsatellite locus Nm=2.610, Fst=0.0874, Fit= 0.4116) indicated that a population and b population exchanged individuals resulting to inbreeding. There was a low level of genetic variabilities between a population and b population. The inbreeding level of b population was higher than that of a population (a population: Fis=0.3221，b population: Fis=0.3983). Therefore, at the present, the focus of management of captive Giant Panda should shift to avoiding the inbreeding and the loss of genetic diversity. The captive populations of Giant Panda should be in the same management unit. Retrieving pedigree and chosing the fittest exchange individual are the key point. Microsatellite technologey is the key way to protect and improve genetic diversity of captive Giant Pandas.]]> 2007/7/5 0:00:00 WANG Fang,PENG Zhen-Xin,ZHANG Jin-Guo,TIAN Liang,HAN Hong-Bing,CAO Zhi-Chen,ZHANG Zhi-He,SHEN Fu-Jun,LI De-Sheng,ZHANG He-Min,ZHANG Li-Bo and LIAN Zheng-Xing 1052true －) on the inactivation and morphological changes of Escherichia coli (E.coli) on the surface of bio-indicator carrier were investigated. The O－ flux was generated from a novel developed O－ generator where the O－ storage-emission material of [Ca24Al28O64]4+·4O－ (abbreviated as C12A7-O－) was used as a pure O－ emitter. Present results showed that inactivation of E.coli was sensitive to the O－ intensity and the cell mortality was enhanced to more than 3-logarithm reduction with the exposure to 1.5 mA/cm2 O－ flux for 120 min. Field emission scanning electron microscopy (FESEM) observations showed that O－ flux destroyed cellular structures. Lipid peroxidation reaction induced by atomic oxygen radical anion for E. coli cells was also observed using product of malondialdehyde (MDA) as an index. The concentration of MDA increased to 1.2 μmol/g(dry weight) of cells when E. coli suspension (5.6×107 cfu/ml) was treated by the O－ flux (1.5 μA/cm2) for 15 min. The findings revealed that the atomic oxygen radical anions, with strong oxidation power, was effective in inactivating E. coli and caused lipid peroxidation reaction at the first time, which would be potential useful to develop a novel approach for the microbial decontamination and for the study on the interactions between microorganisms and O－.]]> 2007/6/8 0:00:00 WANG Lian,SONG Chong-Fu,SUN Jian-Qiu,Youshifumi Torimoto,Masayoshi Sadakata and LI Quan-Xin 1051true Glycine soja) and cultivated (Glycine max) soybean seeds. Two-dimensional gel electrophoresis (2-DE) with immobilized pH gradient (IPG, ranges 4～7) strips was used to separate proteins. On the 2-DE gels stained by Coomassie brilliant blue, PDQuest image software detected about 550 protein spots, of which 10 spots show more than 2.5-fold changes in abundance between wild and cultivated soybeans. These 10 proteins treated by tryptic in-gel digestion were characterized by matrix-assisted laser-desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) and peptide mass finger printings of all were obtained. Soybean UniGene database and NCBI were used to identify proteins, of which 5 proteins were identified. They were soybean agglutinin, seed maturation protein PM24, maturation polypeptide, sucrose-binding protein precursor and trypsin inhibitor p20. The potential functions of these identified proteins were discussed.]]> 2007/7/2 0:00:00 LI Chun-Mei,YANG Shou-Ping,GAI Jun-Yi and YU De-Yue 1050true +/H+ antiporter in vacuolar membranes transports Na+ from the cytoplasm to vacuoles using a pH gradient generated by proton pumps, which could reduce Na+ toxicity. It is uncertain that whether the woody plants have the same mechanism. Through differential centrifugation and sucrose density gradient centrifugation, tonoplast vesicles were isolated from Populus tremula calli broken by blender. After establishing pH gradient by V-ATPase, Na+ could dissipate the pH gradient, which indicates that there is Na+/H+ antiporter in the tonoplast vesicles from Populus tremula calli (Km=11.4 mmol/L). Amiloride could inhibit the Na+/H+ antiporter activity. The antiporter could transport Na+ and K+, the affinity for Na+ is higher. Salt stress decreased Km and Vmax.]]> 2007/6/7 0:00:00 LIU Jing-Jing,ZHANG Xu-Jia and lU Cun-Fu 1049true P < 0.05). Cdk-5 activity was increased by 29% of control, and an increase of PKA activity by 30% of control was also found, both Cdk-5 activity and PKA activity were increased highly compared to normal control(P < 0.01, P < 0.01), furthermore, the immunoreactivity of Tau-1(detection of Ser 198/Ser 199/ Ser202, non-phosphorylated site) was decreased(P < 0.01), the immunoreactivity of PHF-1 sites (detection of ser396/ser404, phosphorylated site) was increased(P < 0.05) in these diabetic rats. After treated the DM rats with roscovitine, a specific Cdk-5 inhibitor, Cdk-5 activity was inhibited to 105% vs control, it was not obviously increased compared to control (P >0.05), but PKA activity remained 127% vs control, it was high compared to control(P < 0.01). Moreover, the hyperphosphorylation of tau at Ser198/Ser199/Ser202 epitopes was reversed (P < 0.01), but the level of phosphorylation of tau at Ser396/Ser404 epitopes was remained highly after roscovitine was administered to DM rats. However, after treated the control rats with roscovitine, Cdk-5 activity was about 104% vs control, and PKA activity was 105% vs control, neither Cdk-5 activity nor PKA activity was higher compared to control(P > 0.05), and the level of tau phosphorylation at Ser198/Ser199/Ser202 and Ser396/Ser404 sites was not increased compared to control too(P > 0.05). These results firstly suggest that the increase of Cdk-5 activity and PKA activity may cause the hyperphosphorylation of tau at Ser198/Ser199/Ser202 and Ser396/Ser404 epitopes in diabetic rat hippocampus, and the intracellular calcium may play a role in this process, however, it is needed to be elucidated in great details.]]> 2007/5/29 0:00:00 QU Zhong-Sen,LI Liang,ZHONG Shi-Jiang,NI Hong,ZHAO Yong-Bo and WANG Qun 1048true 2007/8/9 0:00:00 LIU Yong,WANG Feng,YUAN Lin,HU Hong-Hui and TANG Xiao-Da 1047true de novo synthesis, is a critical therapeutic target in cancer therapy. Previous study has shown that TS was able to bind to its own mRNA in human and E.coli, resulting in translational repression. Zebrafish is the best animal model for vertebrate study. In order to study the regulatory mechanism of zebrafish TS, the enzyme were expressed in E. coli BL21 (DE3) and it was purified to homogeneity. Electrophoretic mobility shift assay (EMSA) was used to detect the interaction of zebrafish TS protein and its own TS transcript in vitro and the results showed that zebrafish TS could bound with its own mRNA specifically. Further study revealed that zebrafish TS was able to interact with its own mRNA in vivo using immunoprecipitation : RT-PCR technique. The results provide evidence that zebrafish may be developed as an useful model for studying the anti-metabolism agents.]]> 2007/9/4 0:00:00 SONG Chun-Xia,NIU Rong-Li,DU Chang-Qing,YANG Shao-Li and LIN Xiu-Kun 1046true Mycoplasma hyorhinis infection in human gastric carcinoma. P37 protein is a membrane lipoprotein of Mycoplasma hyorhinis. It was reported that P37 inhibited cell adhesion and induced tumor invasiveness. To investigate the function of P37 in cancer development, the p37 gene was subcloned into shuttle vector of pAdTrack-CMV. Linearized pAdTrack-CMV-p37 was co-transformed into BJ5183 cells with adenoviral genomic plasmid pAdEasy-1. The identified recombinant DNA was transfected into 293 cells to package adenovirus. From the supernatant and cell lysis, the presence of recombinant adenovirus P37 was proved by PCR. And then BICR cells with this recombinant adenovirus of P37 were successfully infected. RT-PCR and Western blot demonstrated the transcription and expression of P37 in infected BICR cells. And the soluble P37 protein can be secreted into the culture supernatant of infected BICR cells. In the migration assay in vitro, the number of migration BICR cells infected with Ad-p37 was 52.03% more than that of control. The construction of recombinant adenovirus provided a good tool for studying P37 function and its molecular mechanism, which will be further used for in vivo migration and invasion experiments.]]> 2007/6/11 0:00:00 GONG Man-Man,MENG Lin and SHOU Cheng-Chao 1045true E. coli BL-21 (DE3) by means of fusion protein, the soluble 19peptide and 21peptide were obtained from one step chitin affinity chromatograph. By such experiment as MTT assay, cell growth curve, TUNEL assay，flow cytometry，the effect of the H22 ascitic fluid transfevent hepatoma of mice and histopathological slice, the biological activity of 19peptide and21peptide were studied. Experiments in vitro and in vivo identified that 19peptide could inhibit tumor cell proliferation, promote tumor cell apoptosis and stop tumor cell in G0/G1 cycle. It also could inhibit human umbilical vein endothelial cell proliferation to some extend. The anti-tumor activity of 19peptide mainly relied on affecting tumor cell directly and partly on inhibiting vascularization.21peptide inhibited proliferation of human umbilical vein endothelial cell much better than that of tumor cell. It almost could not promote endothelial cell and tumor cell apoptosis.21peptide mainly exhibited indirect anti-tumor activity by anti-angiogenesis. The combination of 19peptide and 21peptide obviously inhibited endothelial cell and tumor cell proliferation and promoted them apoptosis. A combination application could markedly enhance anti-tumor activity, make up the defect of 19peptide, 21peptide alone and bring about synergism. It would be an efficient method for tumor therapy in future, which will lay foundation on its mechanism of action research and clinical tumor therapy.]]> 2007/9/12 0:00:00 WANG Shu-Jing,LIU Xing-Han,JI Yu-Bin and CHEN Ning 1044true Isochrysis galbana, malondialdehyde (MDA) content and activities of antioxidant enzymes (superoxide dismutase and peroxidase) were studied. And the inhibition effects of 4 species of antioxidants on this damage to its cells were also investigated. The results showed that 0.10 mg/L of GI marked decreased cell densities, activities of superoxide dismutase (SOD) and peroxidase (POD), and MDA content increased. With increase of GI concentrations, cell densities and activities of SOD and POD decreased sharply, and MDA content increased obviously. When concentration of GI was 0.30 mg/L, the cell densities, activities of SOD and POD decreased to 0.05, 0.56 and 0.59 folds of the control, respectively, and MDA content increased to as higher as 2.2 folds of that of the control set. L-Ascorbic acid, citrazinic acid, lenediamiaetetraacetic acid disodium salt (Na2EDTA) and 3-tert-butyl-4-hydroxyanisole could decrease MDA content obviously, and marked increased cell densities and activities of SOD and POD. When 4 species of antioxidants were added to the medium, the cell densities, and activities of SOD and POD increased to as high as 1.38～1.90, 1.49～2.12 and 1.55～2.13 folds of that of the control set, respectively, and MDA content decreased by 57.7%～87.9%. It was indicated that the excess active oxygens were produced under growth-inhibitor stress and active oxygen participated in the damage of growth-inhibitor to Isochrysis galbana; the L-ascorbic acid, citrazinic acid, lenediamiaetetraacetic acid disodium salt and 3-tert-butyl-4-hydroxyanisole could effectively inhibit membrane lipid peroxidation and further alleviated algal damage.]]> 2007/6/7 0:00:00 SUN Ying-Ying and WANG Chang-Hai 1043true 2007/5/16 0:00:00 XU Peng,TAN Xungang,ZHANG Pei-Jun,ZHANG Yuqing and WANG Bo 1042true 2007/5/23 0:00:00 ZHU Jieying and TANG Jie 1041true 2007/5/31 0:00:00 ZHANG Yan-Ping,LIU Kai and WU Yu-Ling 1040true + hematopoietic cells and HSCs were successfully isolated from human umbilical cord blood, and 31 HSCs-correlative miRNAs were screened by microarrray. Among them, 22 were low in HSCs, and 9 were high. The result of real time RT-PCR confirmed high expression of miR-520h in HSCs. CFC assays showed that miR-520h promotes differentiation of HSCs into progenitor cells. In conclusion, human HSCs have a set of specific miRNAs that contribute to regulation of HSCs functions, which pave the way for exploring the roles of miRNAs in development of hematopoiectic system.]]> 2007/9/12 0:00:00 SUN Jian-Guo,LIAO Rong-Xia,ZHANG liang,LOU Gui-Yu,ZHOU Du-Jin and CHEN Zheng-Tang 1039true 2007/6/15 0:00:00 WANG Feng-Ze,SHA Li,QIAO Ling,WU Lian-Ying,ZHANG Xiao-Dong and YE Li-Hong 1038true 2007/5/22 0:00:00 XU Jian,YAO Kun,DOU Jie,QIN Jian,XU Wen-Rong,CHEN Yun,YIN Quan-Zhang and ZHOU Feng 1037true 2; 21.2% and 29.3% under hypoxia. The HIF-1α protein were down regulated by siRNA. The pSicoR-HIF1 showed higher interfering efficiency than pSicoR-HIF2. The expression of SDF-1α gene in HIF-1α silencing FLSCs were detected by RT-PCR, fluorescent immunocytochemistry analysis and ELISA. The mRNA and protein of SDF-1α gene were down regulated after silencing of HIF-1α. Therefore, the regulation of SDF-1α gene under hypoxia could be activated by HIF-1α, that plays an important role in the influence on the proliferation and differentiation of stem cell.]]> 2007/4/28 0:00:00 JI Lei,XI Jia-Fei,YUAN Hong-Feng,ZHANG Peng,LIU Yu-Xiao,WANG Yun-Fang,SHI Shuang-Shuang,CHEN Lin,NAN Xue,BAI Ci-Xian and PEI Xue-Tao 1036true Pseudomonas aeruginosus by PCR and recombined with plasmid pUCP18 reversely. The recombinant pUCP18/lasRantisense was verified with restriction analysis, PCR and sequence and was transformed in Pseudomonas aeruginosus. The biological effect of pUCP18/lasRantisense was detected by RT-PCR, NAD method and the assay of pyocyanin. The air tubes of rats were infected by pUCP18/lasRantisense strain and then carried on histopathologic slide check. Expected full length LasR fragment (721bp) can be extended from Pseudomonas aeruginosus gene with PCR technology. And it is consistent with LasR gene of Pseudomonas aeruginosa covered in GenBank (NO. NC_002516). The recombinant plasmid was constructed and transformed into Pseudomonas aeruginosus sucessfully. Compared with the rats which were infected by standard strain, the bronchitis of the rats which were infected by pUCP18/lasRantisense strain was obviously eased. It can be concluded that the antisensenucleic acid of LasR gene can depress the virulence of Pseudomonas aeruginosus and reveal a new target site for treatment.]]> 2007/5/29 0:00:00 ZHANG Ling,ZHOU Jun-Li,LI Jing-Ming and LIAO Fang 1035true 2007/5/29 0:00:00 WANG Zhong-Qun,YANG Yong-Zong,WANG Zuo,REN Zhong,TANG Chao-Ke,LIU Lu-Shan,YI Guang-Hui and YUAN Zhong-Hua 1034true HindⅢ and BamHⅠ site was obtained by PCR, which was cloned into pEGFP-C1. PEG-PEI/ pEGFP-VEGF165 complexes were formed by self-assembly and transfected HUVEc. Transfection efficiency was evaluated by measuring the percentage of cells expressing green fluorecensce protein. The VEGF expression was detected by ELISA, RT-PCR, and the effect of transfection on growth of endothelial cell was evaluated by MTT. The results suggested that the formation of PEG-PEI copolymers could help to reduce the cytotoxicity of PEI. After transfection, the strong expression of green fluorescence protein was observed by fluorescence microscopy. The transfection efficiency was influenced by the number of PEG side chains and N/P ratio. Of all copolymers tested, the transfection efficiency of PEG-PEI(5-25-1) at N/P = 30 reached a maximum, which was much higher than that of PEI. The expression of VEGF protein and mRNA increased significantly, and HUVEc proliferation was accelerated after transfection.These results indicates PEG-PEI copolymers can be used as effective gene carriers for delivery of pEGFP-VEGF165 gene.]]> 2007/10/22 0:00:00 ZHANG Xuan,PAN Shi-Rong,FENG Min,LI Zi-Jun,ZHANG Wei and LUO Xin 1033true Saccharomyces cerevisiae mutants, each with one different gene individually deleted, to isolate genes that may mediate cellular resistance to arsenic. A one-step selection method was used. An aliquot of the pooled yeast library was plated on YPD agar plates supplemented with 3 mmol/L sodium arsenite. The genomic DNAs of the arsenic resistant strains were separately extracted, and amplified by PCR to get DNA fragments with UPTAG. The corresponding deleted genes were identified by comparing the PCR-amplified sequences with the UPTAG sequences from the Saccharomyces Genome Deletion Project. Mutations were identified in 104 genes/ORFs showing resistance to arsenic as compared to the wild type strain. To rule out the possibility that the resistant phenotype of these mutants is a result of arsenic-induced mutation during the screening process, the individual deletion strains from the mutant collection were picked up and tested individually for arsenic resistance using the spot assay. Of the 104 mutants identified in the screen, all exhibited significantly more resistance than the wild-type cells. Among the verified strains, 32 mutants turned out to have stronger phenotype that is resistant to 5 mmol/L arsenite. Five of the 32 mutants (FPS1, TMA20, UPF3, YAL066W, YOR309C) showed resistance to 7 mmol/L arsenite. The phenotype data were mapped onto the regulatory network. Bioinformatic studies of the genes revealed four neighborhoods, including mRNA catabolism, response to stress, histone acetylation, and protein synthesis and catabolism.]]> 2007/6/28 0:00:00 DU Li,ZHANG Xin-Yu,YU Yong,CHEN Jing-Si,LIU Yan,XIA Yong-Jing and LIU Xiang-Jun 1032true 2007/4/13 0:00:00 GUO Lin-Lang,GUO Ying,DERICK LAU and XU Yan-Chao 1031true xynA and xynB from Aspergillus sulphureus, and the DNA fragments composed of 574 bp and 594 bp were amplified by polymerase chain reaction (PCR), respectively. The two PCR products respectively digested with EcoRⅠ/BamHⅠ and BglⅡ/HindⅢ were ligated into multiple cloning sites of pET-28a(+). The resulting plasmid is pET-xynAB, in which xylanase A and B are ligated by a 7-amino acid peptide (GlyGlyGlySerGlyGlyGly). E. coli BL21 transformed with pET-xynAB was induced by IPTG, and a special protein band about 50 ku was detected by SDS-PAGE. The protein purified with Ni-NTA column was denatured by 8 mol/L urea and dialyzed for refolding. The recombinant xylanase showed optimal activity at 50℃ and pH 4.4. The enzyme retained above 75% of its activity at the range of pH 2.4～5.4. The xylanase displayed about 50% retained acitivity after incubating at 80℃ for 30 min. Various metal ions have different effects on activity of the recombinant xylanase.]]> 2007/5/25 0:00:00 LI Yi-Hang,QIAO Jia-Yun and CAO Yun-He 1030true 2007/5/9 0:00:00 YE Jian,LIU Li-Xin,XUE Yuan,QU Jing,GAO Guang-Xia and FANG Rong-Xiang 1029true 2007/8/8 0:00:00 LIU Hua-Gui,LIU Yu-Fang,WANG Xiao-Hua,WU Chang-Xin,ZHAO Xing-Bo and LI Ning 1028true in situ hybridization were used to detect the expression of NGX6 mRNA in the human several common types of cancer. Results showed that expression of NGX6 mRNA in the nasopharyngeal cancer (NPC), lung cancer, gastric cancer and colon-rectal cancer was significantly lower than that in their non-cancer normal tissue (P < 0.05, P < 0.01). Expression level of NGX6 mRNA was evidently lower in the NPC, laryngeal cancer, lung caner and colon-rectal cancer with lymph node metastasis (P < 0.05, P < 0.01). Expression of NGX6 mRNA in the NPC, lung caner and colon-rectal cancer had significant correlation with their clinical stages, which NGX6 mRNA was significantly lower in the clinical stage T2, T3 or T4 that than their clinical stage T1(P < 0.05, P < 0.01). The results suggested that the NPC, lung caner, gastric cancer and colon-rectal cancer had evidently lower expression of NGX6 mRNA, NGX6 mRNA might be used as molecular markers for invasion, metastasis and prognosis of NPC, lung caner and colon-rectal cancer.]]> 2008/6/12 0:00:00 FAN Song-Qing,ZHANG Wen-Ling,ZHOU Ming,PENG Shu-Ping and LI Gui-Yuan 1027true in vitro and in vivo. CTLA4Ig-gene modified BMMSCs have a strong immunosuppressive function cultured in both normal and hepatic differentiation medium and this gene modification improves the biological function of BMMSCs. Recombinant adenovirus containing CTLA4Ig gene was constructed, and transferred into rat BMMSCs in vitro. The transfected BMMSCs, cultured in hepatic differentiation medium containing HGF for 14 days, could express hepatocyte-specific markers including AFP, Alb and CK18, and these differentiated cells from BMMSCs also had the capabilities of glycogen deposition and ICG uptake and excretion, which are hepatocyte-specific functions. Expression of CTLA4Ig in the transfected BMMSCs cultured in the normal growth medium or the differentiation medium was confirmed by RT-PCR, Western blot and fluorescent immunocytochemistry, and the expression at 14 d was weaker than that at 7 d. In the model of MLR, the transfected BMMSCs could suppress immune response, and the suppression was statistically stronger than that of BMMSCs at the same number. Even if cultured in hepatic differentiation medium, the transfected BMMSCs also had the same immunosuppression function. In rat liver transplantation model, the CTLA4Ig-gene modified BMMSCs infused into liver graft after operation could prolong the survival time of the recipients. Adenovirus-mediated CTLA4Ig gene transfer into BMMSCs can not change the potential of BMMSCs to differentiate into hepatocytes, and on the other hand, this gene transfer can improve the immunosuppressive function of BMMSCs, which can broaden the applying space of BMMSCs in the clinical therapy for liver diseases.]]> 2008/6/12 0:00:00 LUO Hai-Ying,WANG Yun-Fang,ZHAI Shu-Sen,ZHANG Yu-Jun,NAN Xue,BAI Ci-Xian,SHI Shuang-Shuang,YU Cheng-Ze,YUE Wen,KONG Wei and PEI Xue-Tao 1026true waf1 was induced and the expression of cyclin B1 was suppressed in KYSE-510 cells, and that the expression of GADD45β and 14-3-3σ were induced and the expression of cyclin B1 was suppressed in OE33 cells. These results were verified by real-time RT-PCR and Western-blot. The comparative effects of all six compounds on the regulation of these gene expressions at the mRNA and protein levels were also analyzed by real-time RT-PCR and Western-blot. The results were shown that p21waf1, GADD45β, 14-3-3σ and cyclin B1 were the target genes which mediated the effects of flavones and flavonols on induction of cell cycle arrest in KYSE-510 cells and OE33 cells.]]> 2008/5/29 0:00:00 ZHANG Qiang and ZHAO Xin-Huai 1025true in vivo cellular and molecular changes in the adjacent cartilage were studied in the rat growth plate injury models. Consisted with the histological changes, both Terminal deoxynucleotidyl Transferase Biotin-dUTP Nick End Labeling (TUNEL) assay and in situ hybridization experiment using Col2a1 probe showed there was the sub-injury cartilage region adjacent to the original injury site. Despite the sub-injury region remained normal cartilage structure and Collagen typeⅩ in the extracellular matrix, the chondrocytes within this region showed the dislocation with the cartilage lacunas, and the strand breaks of cleaved DNA in TUNEL assay. That these chondrocytes didn't express Col2a1 mRNA further confirmed they were dead cells. Along with the degradation of sub-injury cartilage, some fibroblast-like cells presented to the cartilaginous region between the sub-injury region and uninjured cartilage. In situ hybridization experiment for Patched 1 (Ptch1), indicator of Indian Hedgehog (IHH) signaling, indicated these fibroblast-like cells could respond to Hh signaling. These results suggest that the bony bridge formation involves series of changes of chondrocytes and Ihh signaling may be involved in the formation of the transient perichondrium-like structure between the sub-injury cartilage and normal cartilage, and partially contribute to the bony bridge formation. Investigating the underlying cellular and molecular changes after the transphyseal injury will contribute us to explore a prevention treatment in the future clinic.]]> 2008/5/29 0:00:00 JIE Qiang,HU Yun-Yu,YANG Liu,LEI Wei,ZHU Qing-Sheng,LI Ming-Quan,ZHAO Li,LU Rong and WANG Jun 1024true P < 0.05). There was no significant difference in theta band power of EEG between LE and LC groups(P > 0.05). The slope of excitatory postsynaptic potential (EPSP) was significantly increased in LE group compared with that of LC group (P < 0.01). Fluoro-Jade-B showed more neuronal degeneration in LE group compared with that of LC group. The results suggested that TBOA induced damage of glutamate transporters and enhanced the neurotoxicity of status epilepticus, which contributed the synaptic plasticity in status epilepticus rats.]]> 2008/5/8 0:00:00 HAN Da-Dong,QIU Jia-Heng,YAO Yang,ZHANG Tao and YANG Zhuo 1023true wx2 in Toxoplasma gondii was analyzed by bioinformatics. The epitope encoding fragments W2b and W2a were amplified from new gene by PCR, respectively. The single-epitope vaccines pcDNA3-W2b, pcDNA3-W2a and double-epitope vaccine pcDNA3-W2b2a were successfully constructed. Mice in group pcDNA3-W2b, pcDNA3-W2a, pcDNA3-W2b2a and two control groups, pcDNA3 and NS, were injected intramuscularly with pcDNA3-W2b、pcDNA3-W2a and pcDNA3-W2b2a epitope vaccines, respectively. As controls, mice were inoculated with NS or empty plasmid pcDNA3. The induced immune responses were tested by ELISA detecting IgG, and flow cytometry sorting the subsets of T lymphocyte. All mice were challenged with highly virulent RH tachyzoites to observe the survival time. Results display that the level of IgG in sera of mice inoculated with pcDNA3-W2b2a was significantly higher than those in pcDNA3 and NS control group, CD4+T/CD8+ T cell proportion of immunized mice was significantly lower than those in control group. After challenged with highly virulent tachyzoites, the mean survival time of immunized mice in pcDNA3-W2b2a group was significantly longer than groups pcDNA3-W2b, pcDNA3-W2a and control group, indicating that the epitope vaccine from new gene wx2 can induce protective immunity in mice．and the protective efficacy of pcDNA3-W2b2a double epitope vaccine elicits better immunoprotectivity than pcDNA3-W2b,pcDNA3-W2a single-epitope vaccine.]]> 2008/6/19 0:00:00 tankui,ZHANG Qiong,FAN Jiu-Bo,XIE Rong-Hua,JIANG Li-Ping,WU Xiang and SHU Heng-Ping 1022true Arabidopsis has a kind of genes encoding proteins with ARM repeat domains and some of these proteins are known to play important roles in plant development and responses to hormone. An Arabidopsis mutant lfr with a distinct phenotype was got in leaf and flower development. The gene is predicted to encode a protein with ARM repeat domains. In order to study its function and molecular mechanism, recombination expression plasmid pGEX-2TGST∶LFR was constructed and transformed into the host bacteria strain Rosetta. Then IPTG was used to induce the recombinant protein expression in engineering strain. The expression products were detected by 12% SDS-PAGE. The GST∶LFR fusion protein was existed in soluble form with a relative molecular mass 77 ku, which is fit with the molecular mass supposed from gene coding frame. After purification by GST-tag affinity chromatography and electroelution, the fusion protein was used as antigen to prepare polyclonal antiserum in rabbits. After the fifth injection of antigen, the antiserum was obtained and further purified by decreased nonspecific bacteria and GST-tag antibody with method of immuno-precipitation. Western blot analysis showed that the purified antiserum, raised against the recombination LFR protein in rabbits, could react to the recombinant protein expressed in Rosetta specifically. And then the nuclear proteins of Arabidopsis wild type and mutant were extracted and separated by SDS-PAGE. Western blot assays revealed that there was a protein band, with a relative molecular mass 50 ku, indicating that antiserum could react to the native protein expressed in Arabidopsis specifically.]]> 2008/5/8 0:00:00 GAO Ning,WANG Zhi-Juan,ZENG Bo and CUI Su-Juan 1021true 2·) under strong illumination, in which the O2· plays a pivotal role in reactive oxygen species (ROS) metabolism during higher plant photosynthesis. The photosystem of Chlamydomonas reinhardtii is highly similar to that of higher plants. The generation of O2· in C. reinhardtii thylakoid membranes and PS Ⅱ particles has been firstly proved by means of spin trapping-ESR technique and NBT-mediated spectral assay. Referring to the spin trapping-ESR evidences in spinach, the mechanisms of O2· generation both in C. reinhardtii and spinach look identical. Compared to higher plant, C. reinhardtii has several irreplaceable advantages: simpler structure; clearer genetic background and genomic sequencing has been finished; easy to introduce mutations in photosystem. As a result, the study concerning the O2· generation in C. reinhardtii introduces a new promising model for further investigation of ROS metabolism in higher plant photosynthesis.]]> 2008/5/29 0:00:00 YUAN Han,HAN Lu,DU LI-Bo,TIAN Qiu,LIU Ke,DU Lin-Fang and LIU Yang 1020true 2008/5/29 0:00:00 QI Li-Sheng,SU Ji-Guo,CHEN Wei-Zu and WANG Cun-Xin 1019true OsRLK gene was obtained through RT-PCR. The target fragment was subcloned into pET29a, and the recombinant plasmid pET29a-RLK was transformed into E. coli BL21(DE3). The target fragment over-expressed in host strain, and the expression level was about 30% of the total cellular protein. After separated by SDS-PAGE, the target band was excised from the gel, and was used as an antigen to raise the antibody in New Zealand rabbits. After separation and purification of antiserum, the anti-OsRLK polyclonal antibody with the titers of 1∶20 000 was successfully prepared. Western blot analysis showed that the antibody could specifically recognize the expressed fragment in E. coli and the OsRLK protein in root tips from rice. In addition, for the first time, the OsRLK was confirmed as a salt stress responsive protein.]]> 2008/4/11 0:00:00 CHENG Yan-Wei,LI Liang,SHEN Rong,QI Yao-Cheng,LIU Xiao-Yu,WANG Ning and ZHANG Wei 1018true 2+ wave is a chain reaction of intracellular Ca2+ release channels through a Ca2+-induced Ca2+ release mechanism. In cardiac myocytes, Ca2+ wave has drawn much attention because it is found to induce arrhythmia genesis. To investigate the microscopic process of wave propagation, Ca2+ imaging was performed with high spatial and temporal resolution via a laser-scanning confocal microscope combined with loose-seal patch clamp. These observation and analysis revealed that Ca2+ waves originated from a stochastic recruiting of Ca2+ release units (CRUs) by a pioneer Ca2+ spark, which had a low possibility in normal cells. During wave propagation, the ‘waiting’ time that the wave propagate between two neighboring CRUs along propagation direction distributed normally, and cells with a lower speed had a more dispersive distribution of ‘waiting’ time. To study the cause of the randomicity, the wave propagation was simulated with a numerical model. The simulation showed that the intrinsic stochastic open process of CRUs can fully explain the above phenomenon. Increasing the maximal open probability of CRUs reduced the randomness of wavefront propagation and enhanced the average velocity of wave meantime. These experimental and numerical results provided an unequivocal quantification for the stochastic behavior of wave initiation and propagation.]]> 2008/8/20 0:00:00 TANG Ai-Hui and WANG Shi-Qiang 1017true IgH gene, which is transcriptionally silent in fibroblast, was knocked out and the targeted fibroblasts can be used to produce goats with IgH disruption through somatic cell clone. The goat immunoglobulin heavy chain J-Cμ fragment was amplified from the genomic DNA of goat fetal fibroblasts GEF88 through PCR and used as homologous arm to construct an isogenic Positive-Negative Selective targeting vector GTIgH. Then the GEF88 were transfected with the linearized targeting vector GTIgH through electroporation and selected in cell culture medium with 0.8 mg/L puromycin. 1 of the 362 drug-resistant clones was positive for targeting events through PCR screen, and the targeted clone was further confirmed by sequencing and Southern blotting. This suggests that one allele of IgH gene has been successfully knocked out in goat fetal fibroblasts.]]> 2008/4/17 0:00:00 LI Bei,YU Hui-Qing,ZHU Cai-Hong,XIE Fa-Zhan,CHEN Jian-Quan and CHENG Guo-Xiang 1016true P < 0.1. Higher ID leaded to higher probability of success and better growth for UCB-MSCs. Then were cytokines, which could stimulate the growth of UCB-MSCs effectively. Based on the high cell ID, the optimized culture condition was determined with cytokines IL-3 (15 μg/L) and GM-CSF (5 μg/L) added in the traditional medium of MSCs. Then the probability of obtaining UCB-MSCs can be increased up to 90% from 30% with traditional culture method. Moreover, the characteristics of UCB-MSCs were tested by flow cytometric analysis and multi-lineage differentiation identification for osteoblast, chondrocyte and adipocyte. The results showed that the fibroblast-like cells expressed MSCs surface markers of CD13, CD29, CD105，CD166 and CD44 positively and CD34, CD45 and HLA-DR negatively. Meanwhile the cells could differentiate into osteoblasts, chondrocytes and adipocytes similarly to MSCs derived from bone marrow. In conclusion, an efficient protocol have been developed to culture UCB-MSCs by adding cytokines IL-3 (15 μg/L) and GM-CSF (5 μg/L).]]> 2008/4/17 0:00:00 FAN Xiu-Bo,LIU Tian-Qing,HAO Yong-Jie,LIU Yang,MA Xue-Hu and CUI Zhan-Feng 1015true 2008/4/8 0:00:00 CHEN Lan,WANG Shi-Min and CHEN Run-Sheng 1014true Schistosoma japonicum was constructed by phage display technology. Specific SIEA26～28 ku scFv was obtained by screening the SIEA scFv library using the natural molecular candidate vaccine, SIEA26～28 ku. The specific scFv fragment was subcloned to prokaryotic expression vector PET32a and induced the expression of soluble specific scFv in high level. Subsequently, specific scFv as a probe was used to screen the cercaria cDNA library of Schistosoma japonicum to get the coding genes of SIEA26～28 ku molecules. Results showed that specific SIEA26～28 ku scFv with high level expression was achieved. The corresponding gene, ribosomal protein S4 was obtained by initially screening the cercaria cDNA library with the specific SIEA26～28 ku scFv. The obtaining of specific SIEA26～28 ku scFv lays the foundation for further screening and identification of the coding genes of SIEA26～28 ku, the natural molecular candidate vaccine against schistosomiasis.]]> 2008/4/8 0:00:00 HE Zhuo,WANG Shi-Ping,XIAO Xiao-Qing,ZENG Shao-Hua,LIU Ming-She,LI Lin and ZHOU Shuai-Feng 1013true 2008/6/12 0:00:00 ZHANG Jia-Jun,WANG Jun-Wei,YUAN Zhan-Jiang and ZHOU Tian-Shou 1012true Helicoverpa. armegera screened from the cosmid library of Xenorhabdus. nematophilus BP. Comsmid XnBP83 was sequenced using a strategy of subcloning and primer-walking DNA sequencing. The inserted sequences of XnBP83 have 38 939 bp including 5 ORFs that are associated to the insecticidal activity: xptA1, xptB1, xptC1, xptA2 and xptD1. The results of sequence analysis showed that: 1) The xptD1 of XnBP83 is incomplete, and shares 99% predicted amino acid sequence identity to that of X. nematophilus PMFI296. 2) The xptA1 of XnBP83 has 7 569 bp nucleotides, encoding 2 520 amino acids which shares 98% identical amino acid residues with that of PMFI296. 3) The xptB1 has 3 051 bp nucleotides, encoding 1 016 amino acids. The XptB1 sequence shares 98% identical amino acid residues with XptB1 of PMFI296. There are 28 different amino acids from 620 to 650 amino acids. 4) The xptC1 of X. nematophilus BP has 4 225 bp, encoding 1 408 amino acids. The XptC1 sequence shares 96% identical amino acid residues with that of X. nematophilus PMFI296. There are 18 different amino acids from 627 to 646 amino acids. The amino acid sequence with TAQRYLAK was inserted to downstream to 232nd amino acid. And 5) The xptA2 has a length of 7 574 bp, encoding 2 524 amino acids. The XptA2 sequence shares 90% identical amino acid residues with that of PMFI296. Two distinctly divergence amino acid regions present from 788 to 855 and from 1 630 to 1 784 amino acids. Oral bioassay of the supernatant and cell sediment of XnBP83 against H. armegera, S. exigua, S. litura, and T. ni showed that XnBP83 had broad-spectrum insecticidal activity.]]> 2008/6/12 0:00:00 LI Mei,QIU Li-Hong,WU Guo-Feng,LIU Chang-Kun,SHEN Zhi-Hua and PANG Yi 1011true in vitro and transformed by Epstein-Barr virus (EBV). VH and VL genes were reamplified by PCR and combined to single-chain fragment of variable region (ScFv) genes. ScFv genes were cloned into vector fuse5 and transformed into MC1061 by electroporation to construct the ScFv-displaying phage library. The library was subjected to three rounds of positive and negative cell panning and enrichment，then was secreened by phage-ELISA. The binding specificity of phage antibodies with hepatoma carcinoma cells was confirmed by immunohistochemistry with cultured cells and tissue sections. Detection of ELISA showed that 4 liver cancer patients'B cells transformed by EBV could produce specific antibodies to hepatoma carcinoma cell. 6 types of VH genes and 9 types of VL genes were obtained by PCR reamplification then connected with (Gly4Ser)3 linker to form 54 types of ScFv genes. ScFv genes digested with SfiⅠwere cloned into vector fuse5 and transformed into MC1061 via electroporation. Phage antibody library with sink size being 1.0×108 was obtained through tetracycline-resistant secreening. The percentage of full-length ScFv gene inserted into phage DNA was 80%. The library was subjected to three rounds of positive and negative cell panning and enrichment，then was selected by phage-ELISA. After primacy test by HepG2 cell ELISA,179 clones of phage antibody with positive ELISA reaction were picked out of 533 clones.The percentage of positive clones was 33.6%.Though further screened by a panel of cultured cells ELISA,the clone A82 was found to react strongly with HepG2, HEK293, but not with other human tumor cell lines.It also reacted weakly with human hepatic cell line QSG-7701. The results of immunohistochemistry with cultured cells were same as the results of ELISA. A82 was further analyzed after the DNA sequencing.The sequence of A82 was identical. The length of A82 was 742 bp. The VDJ regions of A82 belonged to VH3-23-D2-21-JH6-linker-V4-2-JL2. It can be concluded that fully human anti-hepatoma single-chain Fv fusion phage libraries with sink size being 1.0×108 was constructed by means of phage antibody library technique in combination with in vitro immunization method and EBV transformation technique. By cell ELISA and immunohistochemistry with cultured cells and tissue sections,the clone A82 was confirmed to specific bind with hepatoma carcinoma cells.The ScFv fragment against hepatoma may be further developed and applied to clinical diagnosis and therapy.]]> 2008/6/12 0:00:00 XIAO Yan,LI Guan-Cheng,LI Yue-Hui,HUANG Jian,TONG Yong-Qing and ZHANG Zhi-Jie 1010true 2008/4/17 0:00:00 LIU Dong,WANG Yue-Xiang,HU Jing-Ying,SUN Shu-Na and SONG Hou-Yan 1009true via regulating the neural-specific alternative splicing of genes as GluR-B.]]> 2008/1/8 0:00:00 PENG Zheng-Yu,Lee Shu-Chen,ZHANG Wei and CHEN Xian-Hua 1008true 2008/1/8 0:00:00 LIU Yu-Xiao,JI Lei,YUAN Hong-Feng,CHEN Lin,XUE Jun,GUAN Zhao-Xuan,NAN Xue,BAI Ci-Xian,WANG Yun-Fang,YUE Wen and PEI Xue-Tao 1007true P < 0.01). After blockade of RNA transcription with actinomycin D, total RNA was extracted from stably transfected HEK293 cells with an interval of 2 h, and the stability of GFP-AUR and GFP mRNAs was analyzed by Northern blot. The results indicated that the stability of GFP-AUR mRNA is less than that of GFP mRNA. These results suggest that within the AU-rich region of 3′-UTR of SDCT2β mRNA there is a negative regulation region which can decrease the stability of mRNA, accelerate the degradation of GFP mRNA and may play a negative regulation role for gene expression at post-transcriptional level.]]> 2008/4/17 0:00:00 BAI Xue-Yuan,CHEN Xiang-Mei,FU Bo and WANG Yang 1006true 2 and 95% atmospheric air. Proliferation of osteoblasts were evaluated by WST-1 assay before and 16 h after HBO on day 1,2,3,4,6,8,10. The cytotoxic effect of HBO on osteoblasts was assessed by a toxicology assay kit. For differentiation study, osteoblasts were seeded in 96 well plates at a cell density of 10 000 cells per well. After 3 days normal culture, medium was changed to osteogenic medium. Subsequently, cultures were exposed daily to HBO of 2.4ATA for 90 min and 1.5ATA for 90 min up to 13 days. Mineralization was evaluated by calcium deposition assay, alkaline phosphatase (ALP) activity and von Kossa staining. To assess the effect of pressure on cell proliferation and differentiation, hyperbaric air treatment was observed in this study. It showed that HBO treatment promotes proliferation of osteoblast in the presence of 10% foetal calf serum (FCS). No significant change in extracellular LDH activity before and after HBO treatment. The study of differentiation demonstrated that HBO enhanced differentiation associated with increased bone nodule formation, calcium deposition and alkaline phosphatase activity. These result suggests that HBO treatment significantly stimulated osteogentic differentiation, which implies a potential application of HBO in bone tissue engineering.]]> 2008/5/29 0:00:00 WU Dong,WU Jian-Shan,YAN Fu-Hua and CHEN Jiang 1005true PRMT5 gene was cloned into vector pGEX-4T-1, resulting in only low expression levels in Escherichia coli (E. coli). Here, it was showed that the several N-terminal amino acids deletions could result in a significant increase in the amount of soluble fraction, while one of them did not affect the protein-arginine methyltransferase activity. And it was also found that the N-terminal 15 amino acids region of PRMT5 may be important for the catalytic activity.]]> 2008/4/8 0:00:00 SUN Li-Tao,ZHOU Zhong-Wei,XIE Xiao-Dong and BAO Shi-Lai 1004true -/- mice bone marrow, 12 male apoE-/- mice, with 8 weeks old,were randomly divided into two groups, AMD3100 group (2.5mg / (kg·2d)) and control group(PBS,0.1 ml/2d). After feeding western (high fat and cholesterol) for 12 weeks, the bone marrow cells were isolated and cultured by way of differential-speed-adherence and Micropore-Method. CD133+ VEGFR-2+ bone marrow cell was identified as endothelial progenitor cells by immunofluorescence. The proliferation, migration and adhesion of endothelial progenitor cells were detected by MTT chromometry, transwell and adhesion test, respectively. By counting the typical endothelial progenitor cells-colony forming units (EPC-CFUs) and observing the size and cell density of second EPC-CFUs, the clonality of endothelial progenitor cells was determined. The expression of CXCR4 mRNA and protein were measured by RT-PCR and Western blot. As a result, the proliferation, migration, adhesion and clonality of endothelial progenitor cells derived from AMD3100 group were attenuated in comparison to the control group; the expression of CXCR4 mRNA and protein of AMD3100 group were also lower to control group. It can be concluded that, lasting administration of AMD3100 inhibits the proliferation, migration, adhesion and clonality of bone marrow endothelial progenitor cells and down-regulate the expression of CXCR4 on endothelial progenitor cells.]]> 2008/4/8 0:00:00 WANG Zuo,ZHOU Xiao-Feng,WANG Ren,TONG Zhong-Yi,JIANG Zhi-Sheng and WANG Gui-Xue 1003true IK) and produced a hyperpolarizing shift in the activation-voltage curve. The responses to PDBu were inhibited by lithium carbonate (50 μmol/L). Further studies showed that when pretreated with MEK/ERK inhibitor U0126 (20 μmol/L), although PDBu significantly reduced IK, lithium did not reverse the effect of PDBu. Thus, the results suggested that PKC signaling cascades, along with MAPK (mitogen-activated protein kinase) pathway, were required in the phosphorylation of potassium channel, which was presented by regulation of potassium channel characteristic. AC-cAMP and their cross-talk with GC-cGMP pathway could also modulate the effect of lithium on PKC activation, which could be one of underlying mechanisms likely related to neuroprotective effect of lithium.]]> 2008/4/17 0:00:00 JIAO Guo-Hui,LIU Zhao-Wei,ZHANG Tao and YANG Zhuo 1002true in vitro with different concentration AGE modified human serum albumin (AGE-HSA) which had been treated with soluble receptor for advanced glycation end products (sRAGE) or not. IL-8 levels in the supernatant were determined using Liquid Chip method. RNA was extracted from the cells and RT-PCR was performed to determine the mRNA expression levels of IL-8 in each group, and GAPDH levels were served as reference during this process. The results showed that AGE-HSA increased IL-8 secretion as a dose- and time- dependent manner. Unmodified HSA had no such effect. AGE-induced IL-8 secretion was significantly inhibited by sRAGE as a dose-dependent manner, and the decrease extent was 82.4% when the cells were incubated with 500 mg/L of sRAGE. AGE-RAGE interaction upregulated IL-8 mRNA expression in HUVEC, and this effect could be blocked by pretreatment of AGE-HSA with intact sRAGE. It was proved that AGE modified protein increases the secretion of IL-8 by endothelial cells through RAGE on protein and gene levels, which maybe provide a new aspect to study the machines and treatment methods of AGE-associated diseases.]]> 2008/5/29 0:00:00 ZHAO Shan-Chao,LIU Jing-Hua,SONG Xian-Lu,GUO Zhi-Jian,DENG Peng,LIU Zhi-Qiang,ZHANG Xun,HOU Fan-Fan and JIANG Yong 1001true 2O3 is a Chinese traditional medicine. In previous study, 1.0 μmol/L As2O3 have been found that could induce MG-63 cell apoptosis in vitro. And then it was identified that IEX-1 gene expression was down regulated after the addition of 1.0 μmol/L As2O3 using cDNA Array, RT-PCR, Northern blot. IEX-1, a recently discovered early response gene, regulates cell growth and apoptosis. The exact transcriptional mechanism of IEX-1 was found using Dual-luciferase assay, EMSA, Western blot. Transcriptional factor p53 was up-regulated after the addition of 1.0 μmol/L As2O3, and the increased p53 protein bind the promoter region of IEX-1 gene, which repress the transcription and result in down-regulation of IEX-1 gene expression. It is further confirmed that IEX-1 gene is associated with osteosarcoma and the exact transcriptional mechanism of IEX-1 after the addition of As2O3 was found.]]> 2008/4/17 0:00:00 XIAO Tao,ZHU Wu,YAO Mao-Jin,FANG Jian-Zhen,LUO Yuan-Ming,ZENG Wei and HU Jin-Xi 1000true ®-technology was put in use to study some change of mouse mammary epithelial cells after miR138 inhibitor. It was shown that miR-138 suppresses the exepress of PRL-R(P < 0.05). It was proved that miR-138 can inhibit activity and proliferation of mouse mammary epithelial cells after miR-138 inhibitor(P < 0.01). HPLC was used to study expression of β-casin mouse mammary epithelial cells after miR-138 inhibitor, it shows down express of β-casin(P < 0.05). Results indicated that the target of miR-138 was PRL-R, miR-138 suppresses translation of PRL-R. MiR-138 suppressed the viable and proliferation of mouse mammary epithelial cells.]]> 2008/4/17 0:00:00 WANG Chun-Mei,LI Qing-Zhang and LI Ye 999true lox mediated DNA recombination in embryos and somatic cells, a working model was established using rat mammary carcinoma cell line SHZ-88, aimed at creation of and use repeatedly of selected “friendly loci” in transgenic livestock. An integration vector pTE-lox2272-DsRed-loxP-GFP-loxP, which red fluorescence gene DsRed served as the first target gene and green fluorescence gene GFP as marker gene, was constructed for introduction of acceptor loci in genome. At the same time a replacement vector pT-lox2272-neo-loxP in which Neo coding sequence served as the second target gene was also constructed for replacing DsRed gene. Transgenic cell clones were produced by electroporating SHZ-88 cell with the integration vector. Cells from three transgenic clones selected randomly were further amplified and were then co-electroporated with the replacement vector as well as cre gene. Analysis of the expression patterns of DsRed and GFP indicated that among the 1 070 cell colonies the efficiency on marker GFP deletion was 91.1% and the efficiency on gene replacement was 29.3%. Molecular analysis by PCR and Southern blotting confirmed that the color patterns as expressed by cell colonies could represent the actual molecular events. This working model mediated by Cre/lox system should be useful for the improvement of the present animal transgenic technology.]]> 2008/4/17 0:00:00 CUI Wen-Tao,REN Li-Ming,HOU Jian,ZHANG Ying,CHEN Yong-Fu and AN Xiao-Rong 998true O-tetradecanoyl phorbol-13- acetate (TPA) response elements (TRE) might be located on the -152～-60 position of the 5′ flanking region of the promoter. However, the nucleic proteins bound to the elements have not been identified. In present study, the TRE binding-proteins from EC109 cells following TPA induction were purified by oligonucleotide trapping DNA affinity chromatograph, further separated by SDS-PAGE and stained with silver staining. And then the protein bands were subsequently identified by MALDI-TOF-MS and the resulting spectra were searched to NCBI protein data bank through Mascot. C19, KIAA1949, TDRD1, RXRβ, FAM54A, KLF15, KLF10 and YY-1 eight proteins were identified according to their function, localization and molecular mass. Finally, RT-PCR was performed to determine the TPA responsiveness of these nucleic proteins. Results showed that C19, KIAA1949, TDRD1, RXRβ and KLF15 proteins have the obvious TPA responsiveness in the transcription level. The results suggest these proteins might respond the stimulation of TPA and regulate the expression of NGAL in EC109 cells.]]> 2007/12/19 0:00:00 MENG Ling-Ying,MIAO Cheng-Gui,DU Zeng-Peng,CAI Wei-Jia,XU Li-Yan and LI En-Min 997true in vitro, osteoblast-like cells ( MC3T3E1) growth on the substrates coated with nacre were compared with that growth on the substrates coated with hydroxyapatite (HA) or nothing and the cells of the blank. MC3T3E1 were incubated on the respective surface for 3days, 5days and 7 days. Cell morphology was estimated by inverted phase contrast microscope and scanning electron microscope (SEM) and cell proliferation was measured by flowcytometry. The King's chromometry was used for measuring the activity of ALP in the cell lysate and the expression of TGF-β1 was detected by Western blotting. The results showed that MC3T3E1 cultured on the nacre-coated surface were more spreaded and plumper than the controls on the 3days, 5days and 7days of culture. Meanwhile, the test group showed a superiority to control groups and blank with respects to the cell proliferation index[(35.9±2.5)%, (69.7±3.3)% , (58.2±2.6)%], ALP activity [(6.123±2.917), (17.486±1.986), (23.987±1.372) U/g] and TGF-β1 level. In conclusion, the surface coated nacre can promote the proliferation and differentiation of osteoblast, which the mechanism partly may be the increasing expression of TGF-β1.]]> 2007/12/18 0:00:00 ZHU Li-Qin,WANG Hui-Ming,XU Jun-Hua,WEI Dong,ZHAO Wen-Quan,WANG Xiao-Xiang and WU Nan-Ping 996true +CD25+ regulatory T cells in vitro. A fluorescein-labeled monoclonal antibody staining combined with flow cytometry was applied to detect the effect of different doses of Jagged-1/Fc on the differentiation of the lymphonode cells into CD4+CD25+ T cells at different time, and to measure intracellular cytokine changes of the T cells induced by Jaggde1/Fc. The level of TGF-β1, IL-4 and IL-10 secreted by the T cells that were induced by Jagged-1/Fc was determined by ELISA. The results showed that over 500.0 μg/L of Jagged-1/Fc led to the obvious enhancement of the proportion of CD4+CD25+ T cells within the day 4 to 6 of induction, which was abrogated with an anti-Jagged-1/Fc monoclonal antibody. This induction action of Jagged-1/Fc on CD4+CD25+ T cells was also inhibited by the block of a Notch signal pathway with N-[N-(3,5-difluorophenacetyl)-L-alanyl]-S-phenylglycine t-butyl ester (DAPT). The level of IL-4 and IL-10 in the supernatant of T cell culture and their intracellular level were elevated by the induction of Jagged-1/Fc，and the level of TGF-β1 in the supernatant was not altered. These findings suggest that a soluble Jagged-1/Fc chimera protein may induce the differentiation of mouse lymphonode cells into CD4+CD25+ regulatory T cells in vitro.]]> 2007/12/28 0:00:00 XING Fei-Yue,LIU Jing,YU Zhe and JI Yu-Hua 995true LCRG1) is a candidate tumor suppressor gene of Laryngeal carcinoma. To further investigate its transcriptional regulation, the transcriptional start sites for LCRG1 gene have been identified by 5′ RACE (rapid amplification of cDNA ends) based on the bioinformation analysis of LCRG1. Then eleven luciferase expression vectors which contained potential human LCRG1 gene promoter were constructed. Luciferase reporter assay indicated that LCRG1 promoter region was mainly located in －169～＋127 region nearby the major transcriptional start site. These results suggested that the region (－169～＋127) includes an essential promoter for human LCRG1 gene transcription.]]> 2008/2/3 0:00:00 XIE Hai-Long,CHEN Zhu-Chu and LI Jin-Hua 994true Riemerella anatipestifer were compared by a proteome analysis. Three protein spots differentially expressed between the two strains were observed by 2-DE gels, and were further analyzed using in gel tryptic digestion and peptide mass fingerprinting. Three proteins were identified. W1 was Hsp20, W2 and W3 were transposon. Although the exact role of these proteins has not been characterized, the exclusive expression in virulent strain may indicate that they play an important role in the pathogenesis of Riemerella anatipestifer infection. Although only two virulence factors identified, it opens a path to the further analysis of virulence factors of Riemerella anatipestifer.]]> 2007/12/19 0:00:00 LIU Yan,WEI Qiang,BAO Guo-Lian and JI Quan-An 993true 2-integrin (Mac-1) play a major role in the process. Recently, it was reported that RO-heparin can inhibit selectin-mediated leukocyte adhesion. The effect of RO-heparin on the Mac-1-mediated neutrophils adhesion were further tested. The results showed that RO-heparin could effectively inhibit neutrophils binding to ICAM-1, adhering to COS-7 cells expressing human ICAM-1, and adhering to human umbilical vein endothelial cells (HUVECs) under static and flow conditions. The findings suggest that the effect of RO-heparin on leukocyte adhesion is mainly due to its inhibition on the interaction between selectins or Mac-1 and their ligands and that RO-heparin might be useful in preventing inflammation diseases.]]> 2008/2/3 0:00:00 CHEN Zhi-Hong,JING Ya-Jie,WANG Xiao-Guang and ZENG Xian-Lu 992true 2+) sensor that exists in all eukaryotes . Calmodulin-binding proteins (CaMBPs) play important roles in various signal pathway of calmodulin. The finding of new CaMBPs will be useful for illustrating mechanism of CaM implicating in plant growth and development. Yeast two-hybrid system is an effective method for studying protein-protein interactions in vivo. In the study of plant signal transduction, many important signal transduction molecules were obtained by this system. Here, Arabidopsis flower were used as the material. The Yeast two-hybrid library of Arabidopsis flower was constructed by co-transforming yeast strain AH109 with ds cDNA, pGADT7-rec and pGBKT7-ACaM2. A positive clone was identified. DNA sequencing and analysis indicated that this cDNA clone encodes calmodulin-binding protein AtIQD26. The AtIQD26 protein contains a plant-specific domain of 67 conserved amino acid residues, referred to as the IQ67 domain(IQD), which is characterized by a unique and repetitive arrangement of three different CaM recruitment motifs, known as the IQ, 1-5-10, and 1-8-14 motifs. Yeast two-hybrid analysis and gel overlay experiments demonstrated that AtIQD26 interacted with CaM both in the presence or absence of Ca2+. A striking feature of AtIQD26 is the high isoelectric point (～10.6) and frequency of serine residues (～10%). To uncover potential roles for AtIQD26, a series of expression vectors were constructed about it, and the relative transgenic works were finished. Using these transgenic lines, the tissue expression and the subcellular localization of AtIQD26 were studied. Fusion GFP reporter showed that AtIQD26 was located at the nucleus and near cytoplasm membrane. GUS histochemical assay showed that AtIQD26 had characteristic of universal tissue expression, especially in the renascent tissue. Interaction of AtIQD26 with CaM and the presence of predicted CaM binding sites in it suggest that AtIQD26 is a new member of CaM targets. The basic isoelectric point and its potential nuclear localization suggest that AtIQD26 links Ca2+ signaling pathways to the regulation of gene expression. Expression similarity indicates that AtIQD26 combined with CaM to regulate plant development and growth.]]> 2007/12/27 0:00:00 WEI Hui-Yan,GUO Zhen-Qing,WANG Zhen-Jie,LI Zhao-Wei,WANG Zhi-Juan and CUI Su-Juan 991true 2), blood glucose level decreased, HIF-1α expression increased and myocardial infarction size reduced in GK diabetes rats, which may provide a new insight on treatment of coronary heart disease combined with diabetes by regulating HIF-1 signal transduction system.]]> 2008/4/16 0:00:00 WANG Yu-Jie,ZHANG Shi-Guang,ZHENG Xiao-Wei and QI Guo-Xian 990true 2008/6/12 0:00:00 WANG Jian,ZHU Si-Bo,LI Ding and ZHU Nai-Shuo 989true 2008/2/3 0:00:00 CHENG Ai-Lan,HUANG Wei-Guo,ZHANG Peng-Fei,LI Mao-Yu,PENG Fang,LI Feng,LI Cui,YI Hong,LI Mei-Xiang,CHEN Zhu-Chu and Xiao Zhi-Qiang 988true 2007/11/20 0:00:00 ZHANG Song,XIA Xue-Feng,SHEN Jin-Cheng and SUN Zhi-Rong 987true Pneumococcal surface protein A (PspA) and its conjugates with polysaccharide were researched in mice injected with PspA and its conjugates respectively. The enzyme-linked immunosorbent assay (ELISA) was adopted to detect the immunogenicity of the antigen. Passive and active protection experiments against intraperitoneal challenge with Pneumococcal pneumonia were carried out to validate the protection of PspA and the conjugates with polysaccharide. The experimental results showed that the immune effects of both the conjugates and PspA were effective to against invasion of Pneumococcal pneumonia in mice (P < 0.01). And the conjugates of PspA with polysaccharide gave the best protection and survived 3 days longer than that of PspA. The conjugates with polysaccharide enhanced the higher level of both IgG and IgG2a antibody titer than that of PspA. The cross-reactivity of Western-blotting showed conjugates with polysaccharide caused immune reaction in different serological Pneumococcal pneumonia, this meant that the conjugates had some cross-protection function against different serotypes 6B, 5, 1, 23F, 19F. The conjugates of PspA with polysaccharide showed better immunocompetence and immune protection against invasion of Pneumococcal pneumonia than PapA and the capsule polysaccharide. This study demonstrated that the conjugates could enhance protection against Pneumococcal pneumonia, which should be pertinent to future efforts to develop new protein-based complex vaccines.]]> 2008/1/8 0:00:00 HUANG Jin-Zhong,LIN Hai-Ying,CAI Qian-Ying,GUO Yang-Hao and Meng Chun 986true 2008/2/3 0:00:00 LIU Lu-Shan,CHENG Yan-Li,XIE Min,YANG Qiong,PAN Li-Hong,JIANG Zhi-Sheng,TANG Chao-Ke,WEI Dang-Heng and TANG Zhi-Han 985true 2007/12/18 0:00:00 REN Wen-Ke,XU Hai-Song and LI Xiao-Qin 984true I 5～9, M 10～100 ku), were found to participate in the metabolism and play crucial roles in binding and catalytic reactions. Putative N-linked glycosylation consensus sequence, i.e., -N-X-S/T-, was applied to evaluate glycosylation of all the identified proteins. Furthermore, protein immunoprecipitation combined with lectin blot was used to further validate fucosylation of the identified possible candidate proteins, haptoglobin and alpha enolase. All the results suggested that lectin affinity chromatography and 2-DE in combination with MALDI-MS/MS enabled the identification of all the specific subsets of glycoprotein, and the database could give us a help when searching for some aberrantly core-fucosylated glycoproteins associated with some diseases.]]> 2008/4/16 0:00:00 DAI Zhi,FAN Jia,ZHOU Jian,BAI Dou-Sheng,QIU Shuang-Jian,KANG Xiao-Nan,CUI Jie-Feng,GUO Kun,LI Yan and LIU Yin-Kun 983true Tachypleus tridentatus, showing wide-spectrum antibacterial activity. However, its mechnism of killing bacterium is not very clear. The aim was to investigate the targets of tachyplesin for further studying the antibacterial molecular mechnism. In vitro bacterial inhibition method was used to determine feature of bacterial inhibition dynamic of tachyplesin. Inorganic phosphate measurement and ultraviolet absorption methods were used to observe the leakage of inorganic phosphorus and large mass molecular before and after bacterium were incubated with tachyplesin. Scanning electron microscopy (SEM) and transmission electron microscopy (TEM) were adopted to investigate the morphologic and structural changes of bacterium before and after being incubated with tachyplesin. Ultroviolet absorption method and electrophoretic mobility shift assay (EMSA) were used to investigate the effect of tachyplesin on structure of genomic DNA and plasmid DNA of bacterium. Plasmid transformation method was applied to observe the effect of tachyplesin on copy and transcription of plasmid DNA. The results showed that, (1) Tachyplesin had different feature of bacterial inhibition dynamic in Gram positive and negative bacterium. In the Gram positive bacterium (E. coli K88 and E. coli F41), the antibacterial activity of tachyplesin increased sharply during the first 10 h, and was in the plateau phase during 10～30 h, and then decreased slowly. In the Gram negative bacterium (B. subtilis W B800 and S. aureus), the antibacterial activity of tachyplesin increased sharply during the first 3 h, and decreased during 3～10 h, and increased again during 10～15 h, the trend was similar to the Gram positive bacterium after 20 h. (2) Bacterium lost inorganic phosphorus and large mass molecular. Phosphorous concentration in the culture media of S. aureus was significantly higher after treated with tachyplesin, the high concentration could keep long time (30 h later). The concentration of the UV absorbing substance in E. coli K88 became higher and higher depending on tachyplesin concentration and time. At low tachyplesin concentration, the UV absorbing substance concentration kept low. (3) The structure of cell wall, cell membrane and the whole cell body were damaged to some extent after treated by tachyplesin. (4) Tachyplesin could combine with genomic DNA and plasmid DNA of bacterium, high concentration of tachyplesin probably could break down DNA, and inhibited copy and transcription of plasmid DNA. It was suggested that the antibacterial targets include at least cell wall and membrane and DNA of bacterium.]]> 2007/12/28 0:00:00 DAI Jian-Guo,XIE Hai-Wei,JIN Gang,ZHANG Yan,ZHU Jun-Chen and GUO Yong 982true in vitro and in vivo. The results showed that NGX6 re-expression considerably suppressed the growth of xenografts in the nude mice. Data from Western blot revealed that NGX6 significantly down-regulated the expression of EGFR, K-ras, p-JNK, c-Jun and cyclin D1 in the colon cancer cells. And the further analysis by immunohistochemistry and Western blot in vivo indicated that NGX6 decreased the expression of EGFR, K-ras, p-JNK, c-Jun and cyclin D1 in the xenografts in nude mice，which were consistent with in vitro observations. Such evidences suggest that the major mechanism of NGX6 in colon cancer is negatively regulating EGFR-mediated JNK pathway, laying the experimental basis for further elucidating the mechanism of NGX6 gene.]]> 2008/1/25 0:00:00 WANG Xiao-Yan,SHEN Shou-Rong,LIU Fen,PENG Ya,LI Gui-Yuam and FAN Song-Qing 981true E. coli ML-35p determined by agarose radial diffusion assay. The N-terminal amino acid sequence of HCP-21 was PKRKAEGDAK and its molecular mass was 9 263.62 identified by amino acid sequencing and mass spectrometry analysis. HCP-21 showed 100% identity to HMGN2 (high mobility group protein N2, HMGN2) fragment 2～11 by GenBank BLAST searching, and had the same molecular mass as HMGN2. So it was certain that HCP-21 was HMGN2. HCP-26 was SLPI (secretory leukocyte protease inhibitor, SLPI) fragment by N-terminal amino acid sequencing which was SGKSFKAGVC. It was the same with SLPI fragment 26～35 by GenBank BLAST searching. Total RNA was extracted from primary culture cervical epithelial cells, and the specific primer was designed based on HMGN2 cDNA sequence. RT-PCR showed that a fragment about 270 bp was amplified, which was same to HMGN2 cDNA. It was suggested that cervical epithelial cells could express HMGN2 mRNA in physiological condition. The fusion protein GST-HMGN2 was purified by low-pressure chromatography. The antiserum of HMGN2 was prepared from the immune rabbit with the fusion protein. Cervical tissue paraffin section and cervical mucus smear were detected by immunhistochemistry. The staining results showed that HMGN2 mainly distributed in mucosa surface of cervix and existed in cervical mucus. HMGN2 constitutively expressed in cervical mucosa and mucus, so it might play an important role in innate defensive of cervical.]]> 2007/12/19 0:00:00 WANG Li-Li,HE Yue-Dong,HUANG Ning,LI Ming,XIONG Wen-Bi,FENG Yun,WU Qi,WANG Bo-Yao and PAN Xiao-Ling 980true 2008/4/16 0:00:00 LI Zheng and ZHANG Zhen-Xi 979true 1.75) and 6 proteins down-regulated (Ratio < 0.5). Further research indicated that these proteins were related with liver cancer. Altogether, the results indicated that the strategy was operable and convenient with high sensitivity and wild dynamic range, and will be significant to deliver biomarker discovery either in theory or in clinic.]]> 2007/11/8 0:00:00 CHEN Ming,YING Wan-Tao,FANG Qin-Mei,SUN Wei,HE Fu-Chu and QIAN Xiao-Hong 978true 2007/11/20 0:00:00 ZHANG Wen-Jing,YI Bin,YI Hong,ZHANG Peng-Fei,LI Mao-Yu,LI Cui,RUAN Lin,CHEN Zhu-Chu,LI Jian-Ling and XIAO Zhi-Qiang 977true 2007/11/8 0:00:00 HUANG Jia-Liang,GUO Hong and MENG Chun 976true 2007/12/18 0:00:00 CHEN Guang-Wen,WANG Kang-Kai,LIU Ying,TANG Dao-Lin and XIAO Xian-Zhong 975true vs 0.796±0.173, P < 0.05), however, there was no difference in mafa and mafb mRNA level between islet and duct. The expression of mafa, mafb, c-maf in pancreatic duct epithelium, gives more support to not only the roles of the three genes in endocrine development, but also the potential of converting duct epithelium to endocrine cells. Moreover, the higher expression of c-maf in duct than islet suggests that down-regulation of c-maf might help to differentiation and mature of endocrine cells.]]> 2007/10/19 0:00:00 ZHANG Xian-Ling,GU Yan-Yun,ZHOU Wen-Zhong,ZHANG Hong-Li and LI Guo 974true 2007/11/20 0:00:00 ZHAO Hu-Cheng,ZHENG Xiu-Peng,GAO Xiao-Hui and XI Bao-Shu 973true 2007/11/29 0:00:00 ZHENG Jing-Min,LIU Zhi-Hong and LI Lei-Shi 972true via PCR from λ-lysogen DE3, and the gene was cloned into pBABEpuro retrovial vector, a recombinant plasmid named as pT7BABEpuro was constructed and sequenced. Then the pT7BABEpuro and pVSV-G plasmids were cotransfected into GP2-293 packaging cells by liposomese, some pseudotype viruses were ingathered and transfected into IBRS-2 cell under polybrene. The IBRS-2 cell was propagated in DMEM with puromycin. The genome extraction from the cells transfected different times, the T7 RNAP gene was amplified from the genome by PCR, the mRNA of T7 RNAP protein expressed in IBRST7 cells was analyzed by RT-PCR, respectively, the results showed the T7 RNAP gene had been integrated into the chromosome of IBRS-2 cell and expressed stably at high level. To study whether T7 RNAP is of transcriptional activity in the established IBRST7 cell line, a plasmid pIERS-EGFP-ET with a reporter gene (EGFP) under control of the T7 promoter was constructed. IRES element from FMDV (for CAP-independent translation) was cloned into plasmid pET-43.1a-c(+) downstream of the T7 promoter sequence, then EGFP gene was cloned in frame downstream of the AUG codon of the FMDV IRES, resulting in the plasmid. IBRST7 cells were transfected with plasmid pIERS-EGFP-ET using lipfection, EGFP was expressed, the results showed the T7 RNAP in IBRST7 cells has transcriptional activity. IBRST7 cell line was directly transfected with linearized full-length cDNA of swine vesicular disease virus (SVDV) HK/70, infectious SVDV was efficiently recovered from the cDNA. The reverse genetic procedure is simplified to a faster, one step protocol to recover RNA virus and will be useful to understand the mechanisms of molecular pathology of RNA virus and develop effective vaccines.]]> 2008/3/7 0:00:00 ZHENG Hai-Xue,TIAN Hong,JIN Ye,WU Jin-Yan,SHANG You-Jun,Liu Xiang-Tao and XIE Qing-Ge 971true P < 0.05). These data suggest that NOR1 may influence the biology and cancerous behaviors of HepG2 cells by regulating expression of a set of genes involved in signal transduction, cell cycle regulation, transcription and translation controls.]]> 2008/2/3 0:00:00 LI Deng-Qing,TANG Hua,GUI Rong and NIE Xin-Min 970true P < 0.05). The FRET efficiency of BACE-NT/FPs group is (3.80±0.74)%, which is not statistically different from that of control group. These results indicat that the BACE-FL exists as a dimmer in living cells, but the BACE-NT exists as a monomer, suggesting the transmembrane and C-terminus regions are important for normal transport and localization, and is necessary for the dimerization of BACE.]]> 2007/10/15 0:00:00 LU Jin-Ling,CHU Jun,YANG Jie,ZENG Shao-Qun,LUO Qing-Ming and ZHANG Zhi-Hong 969true 2007/8/22 0:00:00 LIU Xiao-Ping,ZHU Fei-Zhou,WANG Guo,DENG Qi-Pan,ZHANG Jing,LI Qiong,LING Jie,WU Ling-Qian,LIANG De-Sheng,PAN Qian,LONG Zhi-Gao,GONG Hui-Yong,XIA Jia-Hui,ZHANG Zhuo-Hua and XIA Kun 968true 1 (FB1) is a fungi toxin, which is a competitive inhibitor of ceramide synthase in de novo sphingolipid biosynthesis. FB1 can induce PCD in both animal and plant cells. To dissect this pathway, a genetic screen for fumonisin B1 resistant (fbr) mutants, and identified 11 fbr mutants was carried out. Genetic analysis showed that these 11 mutants belonged to 9 complementary groups or genetic loci. Here a detailed phenotypic analysis of fbr136 was reported. In addition to the resistance to FB1, fbr136 also showed resistance other PCD-inducing compounds, including H2O2 and paraquat. Furthermore, FB1-induced PR1 expression was reduced in fbr136, suggesting that a PCD pathway is likely impaired in the mutant. When stained with nitroblue tetrazolium (NBT), fbr136 showed a reduced accumulation of reactive oxygen species (ROS) induced by FB1. The fbr136 mutation was roughly mapped onto chromosome Ⅲ, where no other fbr mutants have been previously identified. It is proposed that FBR136 may act as an important regulator in a sphingolipid-mediated PCD pathway, involved in the generation of ROS.]]> 2007/11/7 0:00:00 SUN Shu-Lan and ZUO Jian-Ru 967true 2007/10/19 0:00:00 WANG Liang,YANG Zhi-Hua,GONG Jing-Bo,ZHAO Xiao-Ling and QIAN Ling-Jia 966true 2007/9/12 0:00:00 WANG Jing,HU Qing-Hua,MI Qiang,ZHAO Lei,ZHOU Jun-Nian,QIN Li-Peng,YANG Chun,WANG Yun-Fang and ZHUANG Feng-Yuan 965true in vitro culture. The efficiency of generating newborn ES mice of wild type SCR012 cells was higher (8.3%) than that of CJ7 (2.7%). In the meanwhile, adult and fertile mutant ES mice with two gene knock-out ES cells-18 KO (derived from CJ7) and F8 KO (derived from SCR012) were generated. Interestingly, the generation frequency of 18 KO ES mice was even higher (8.1%) than wild-type CJ7 ES mice, indicating that the development potential among ES cell clones was significantly different from each other and the decline of development potential was not equal among ES cell clones during ES cell passage in vitro culture. The F8 KO mice had indistinguishable bleeding phenotype, which is identical to that of mice derived from breeding of chimeric mice. These results suggested that this technique can be used as a powerful approach to produced gene targeted mice efficiently.]]> 2007/8/20 0:00:00 KUANG Ying,SUN Xia,DENG Tao,LU Xi-Bin,SUN Rui-Lin,WANG Zhu-Gang and FEI Jian 964true 9 to 1∶210. The rescue system of six internal genes used as backbone opens the way for further research on gene function and neotype vaccine candidate of cold-adapted, live attenuated human influenza virus.]]> 2007/9/21 0:00:00 YANG Peng-Hui,YE Yi,SHI Xin-Fu,LUO De-Yan,ZHANG Yu,YAN Yan,ZHANG Peng-Fei,DUAN Yue-Qiang,XING Li,LIU Xiu-Fan and WANG Xi-Liang 963true 125I-rT3 as substrate combined with ion-exchange chromatography. The thyroid hormone was measured with radioimmunoassay method. The data revealed that in the case of iodine deficiency, both D1 mRNA expression and D1 activity was greatly increased(compared with NI groups, P < 0.01). T3 and T4 in thyroid tissue was significantly decreased, but T3 / T4 was increased (P < 0.01). On the other hand, when faced to iodine excess, D1 mRNA expression was reduced (There was a tendency of decreasing in D1 mRNA expression in all HI groups compared with NI group, significant difference was found in 5HI and 50HI at 6 months ), while there was no remarkable effect on D1 activity. The thyroid hormone assay showed that T3 / T4 was decreased. In conclusion, D1 is a critical factor in the regulation mechanism of thyroid function. The up-regulated mRNA expression and activity under the state of iodine deficiency will improve the conversion of T4 to T3 to maintain the normal thyroid function. The inhibition of D1 expression induced by iodine excess may be taken as an effective way to protect organism from impairment caused by too much T3.]]> 2008/1/4 0:00:00 WANG Kun,SUN Yi-Na,LIU Jia-Yu,YAN Yu-Qin and CHEN Zu-Pei 962true P < 0.05), compared with A375-WT cell. FCM analysis indicated that apoptosis cell in A375-G6PDΔ was 2.86 times as that of A375-WT(P < 0.01) with 33.8 % increase of SPF(P < 0.05), 59.7 % raise of PI(P < 0.01), and 27.7 % decrease of G0/G1 phase(P < 0.01). Furthermore, apoptosis-associated protein check showed that Caspase-3 was 2.86 times as that of A375-WT(P < 0.01) with 54.7% descend of P53(P < 0.01). It is proposed that G6PD can maintain the growth and proliferation of A375 cell. G6PD deficiency probably restrains the change proceeding of G2/M phase to G0/G1 phase in A375Δ cell cycle through down-regulation P53 expression and up-regulation Caspase-3 expression. The role and mechanism of G6PD in cell growth, proliferation, and differentiation of tumor cells need to be further investigated.]]> 2008/2/25 0:00:00 ZHU Yue-Chun,LU Hui-Ru,LI Dan-Yi and TONG Shu-Fen 961true 2+-dependent manner, which was partially inhibited by some saccharides (mannose, glucose, or N-acetylglucosamine), C1q and anti-C1qR mAb. Similarly, it was also incompletely inhibited by recombinant human MBL carbohydrate recognition domains (rhMBL-CRD) protein and recombinant human MBL collagen-like region (rhMBL-CLR) protein, respectively, but completely inhibited by both. These data indicated that Raji cells express Ca2+-dependent and sugar-sensitive CLR-specific and CRD-specific MBL receptors, of which, the former was also shared with C1q. Further investigation showed that MBL could significantly inhibit Raji cell proliferation directly at higher concentrations (10～50 mg/L) in a dose-dependent manner, suggesting that MBL might also play certain roles in adaptive immune responses.]]> 2007/10/15 0:00:00 WANG Ming-Yong,ZHANG Ya-Ni,ZHANG Li-Yun,LU Xiao,LEI Ming,LI Rui-Fang and CHEN Zheng-Liang 960true Flavivirus) is an arbovirus of public health importance. The envelope glycoprotein of JEV is associated with viral attachment and fusion with host cell, determine the virus′s hemagglutination ability, cellular tropism, viral virulence, and induction of protective immune response. The domain Ⅲ of envelope protein (E protein) is an important region in inducing neutralizing antibodies against JEV. In order to study the antigenic structure of domain Ⅲ on E protein, domain Ⅲ of the envelope protein was expressed by fusion with GST in a pGEX-6p-1 vector. Western blot demonstrated that expressed fusion protein could be recognized by anti-JEV sera. To map the antigenic epitope of this region, a set of 14 partially overlapping short peptides spanning the domain Ⅲ were designed and expressed in fusing with GST. Then Western blot and ELISA reactivity of these short peptide fusion proteins to anti-JEV sera were surveyed, respectively. Four linear antigenic epitopes, E39 (305TEKFSFAKNPVDTGHG320), E45-1 (355VTVNPFVATSSA366), E48-1 (377PFGDSYIV384) and E49 (385VGRGDKQINHHWHKAG400) were identified. Immunization of mice with epitope fusion proteins revealed that all four proteins could elicit short peptide specific antibody. And in vitro neutralization test verified that E39 was linear neutralizing epitope. This result provides important basis for further structural and functional study of domain Ⅲ of JEV envelope protein, and development of diagnostic techniques and vaccines.]]> 2007/8/28 0:00:00 YAN Li-Ping,HUA Rong-Hong,QI Wen-Bao and TONG Guang-Zhi 959true 2007/10/19 0:00:00 NIE Gan-Juan,ZHOU Jian-Hua,LI Mao-Yu,ZHANG Peng-Fei,DUAN Chao-Jun,LI Cui,YI Hong,TANG Cen-E,FENG Xue-Ping,PENG Fang,CHEN Zhu-Chu and XIAO Zhi-Qiang 958true n = 6, P < 0.01). When the expression of NRSF was down-regulated, genes regulated by NRSF, such as SCG10, BDNF, pax4 and insulin, began to express in HeLa cells. The human insulin promoter activity was up-regulated 2.4-fold after RNA interference (n = 3, P < 0.01), confirmed by dual-luciferase assay.]]> 2008/2/3 0:00:00 YANG Yin-Xiang,LI Yan-Hua,LIU Qing-Bin,ZHANG Sai-Nan,CHEN Lin,LU Yang,BAI Ci-Xian,NAN Xue and PEI Xue-Tao 957true TM 2000. The changes of cyclin D1 expression were detected by fluorescent quantitative PCR(FQ-PCR), semi-quantitative RT-PCR. The effect of sequence- specific small interfering RNA in suppressing the proliferation of fibroblasts was detected by MTT assay. Flow cytometry were used for evaluation of cell cycle. The expression of cyclin D1, CDK4, pRb and P16 was detected by immunohistochemical method. The results showed that: (1) The sequence- specific siRNA effectively suppressed cyclin D1 expression at both mRNA levels with inhibition rate of 63.68% and 92.83% (P < 0.01). (2) Significantly inhibited the proliferation of fibroblasts, and changed cell cycle in percentage of G0/G1 phase cells was increased compared with the controls groups in fibroblasts(P < 0.05). (3) 72 h after transfection, the expression of cyclin D1, CDK4 and pRb decreased, and the expression of P16 increased. It can be concluded that the plasmid expression vector for the RNAi against cyclin D1 constructed in the study can effectively and specifically suppress cyclin D1 expression, and progression of G1/S is effected by G1 phase related regulatory protein, and suppresses proliferation of fibroblasts derived from keloid.]]> 2007/9/12 0:00:00 YU Dong-Mei,HAO Li-Jun,WANG Dong-Yan and WANG Hui-Dong 956true S. trifolia at five different developmental stages by CSEM-EDX technique. At early stage, K and Cl concentrations in cell wall were high up to 36% and 4.3% of dry weight, respectively. It supported the hypotheses that aerenchyma spaces are filled with liquid at early developmental stages of aerenchyma in S. trifolia petiole. Mg concentration was high at stage 2, up to 0.86% of dry weight. Zinc and Cu were detected only at rapid expansion stages, during which the concentrations were up to 1.5% and 2.5%, respectively. Calcium was detected in the cell wall only at mature stages, the concentration was high up to 1.3% of dry weight at stages 4 and 5. These results confirmed that the element concentration of aerenchyma cell wall undergoes dynamic changes during different developmental stages, and a low Ca with high Zn and Cu concentration are needed for cell expansion. Copper and Zn deposition in the cell wall showed a significant positive linear correlation, suggesting that these two elements share same or similar uptake and transport mechanism in plants.]]> 2007/10/19 0:00:00 LIANG Feng,WANG Zhong,GUO Jian-Lin,SHEN Li-Zhen and YANG Qing 955true to) function and maintaining the ventricular electrical stability. The role of two kinds of long QT syndrome (LQT6)-associated mutations in KCNE2, I57T and V65M in regulation of Kv4.3, the major α subunit of Ito was investigated in COS-7 cells to elucidate the mechanism for pro-arrhythmia induced by these two inherited mutations. The results demonstrate that coexpression of KCNE2 with Kv4.3 slowed the rate of activation and inactivation, and caused a positive shift of voltage-dependence of inactivation of Kv4.3 channel. Moreover, KCNE2 accelerated recovery of Kv4.3 channel from inactivation. This is in agreement with previous study． In comparison with the effect of wild type KCNE2, I57T exerted no significant effect on Kv4.3 gating kinetics and recovery property. The phenotype of I57T and Kv4.3 coexpressed channel was similar to that of Kv4.3 expressed alone. In contrast, V65M enhanced all the effects of KCNE2 on Kv4.3, including slower activation and inactivation, more positive shift of voltage-dependence of inactivation and faster recovery from inactivation versus Kv4.3+KCNE2. Furthermore, V65M dramatically decreased Kv4.3 current amplitude. These results suggest that I57T causes “loss of function” while V65M results in “gain of function” of KCNE2. It is therefore proposed that KCNE2 is an important modulator of Ito. LQT6-associated mutations in KCNE2 may perturb the contribution of Ito to electrical stability in the heart through changing KCNE2 regulation of Kv4.3, predisposing the heart to arrhythmia under certain conditions.]]> 2007/8/28 0:00:00 LIU Wen-Juan,DENG Peng,CHENG Wei-Wei,DENG Jian-Xin,PAN Bing-Xing,JIANG Yong and LIU Jie 954true 2007/11/8 0:00:00 ZHOU Jun,LI Jian-Ming,YANG Fa-Da,LIU Yu-Hong and DING Yan-Qing 953true －), which is a highly reactive free radical that has been shown to mediate much of the toxicity of NO. ONOO－ has also been implicated as playing a role in the pathology of various brain disorders. The aim of the study was to investigate the actions of ONOO－ on voltage-gated sodium currents and its effect on membrane excitability in hippocampal CA1 neurons. SIN-1, which leads to the simultaneous generation of superoxide anion and NO, and then forms the highly reactive species ONOO－, induced a dose-dependent inhibition in amplitudes of transient potassium currents (INa). SIN-1(500 μmol/L) delayed the recovery of INa from inactivity, but did not have a marked effect on the activation and inactivation parameters of INa. However, co-treamtent with SIN-1 and Urate (100 mmol/L), an ONOO－ scavenger, had no effect on INa currents. SIN-1 (500 μmol/L) led to the decrease in the amplitude and firing rate of action potential. Pretreatment with adenylate cyclase inhibitor MDL-12, 330A (25 μmol/L) and NEM (50 μmol/L) did not block the effects of ONOO－. On the other hand, the responses to ONOO－ were inhibited by guanylate cyclase inhibitor ODQ (10 μmol/L). The effects of ONOO－ on hippocampal neurons were mediated through activation of cGMP-INa-AP (action potential) signaling cascades and independent from PKA and S-nitrosylation pathway, which maybe one of underlying mechanisms likely related to neurotoxicity of ONOO－.]]> 2007/9/28 0:00:00 LIU Zhao-Wei,HAN Da-Dong,YANG Xi,XU Lan-Ju,ZHANG Tao and YANG Zhuo 952true in vitro model to investigate the influence of hADAM-19 on cell invasion in human trophoblasts. The regulation of hADAM-19 expression by transforming growth factor β1 (TGF-β1) was investigated by RT-PCR and Western blotting. It was shown that over-expression of hADAM-19 in NPC cells led to down-regulation of MMP-9, MMP-2 mRNA expression and decrease in cell invasion. In NPC cells, the expression of hADAM-19 was significantly up-regulated by TGF-β1 in a dose-dependent manner. The data indicated that there existed paracrine regulation of ADAM-19 in human placenta. ADAM-19 was involved in invasion-inhibiting regulation of human trophoblast cells, and it may function, at least in part, through coordination with various MMPs.]]> 2007/9/12 0:00:00 ZHAO Mei-Rong,QIU Wei,LI Yu-Xia,SANG Qing-Xiang and WANG Yan-Ling 951true Arabidopsis thaliana glutathione S-transferases zeta class (AtGSTZ) is a multi-functional enzyme, which plays important role in cellular metabolism and environmental purification. Error-prone PCR and cycles of DNA shuffling were used to construct a mutagenesis library of AtGSTZ. The screening of the resultant libraries was carried out by a pH indicator dye-based colorimetric assay. Nine mutants which enhanced the dichloroacetic acid dechlorination activity were obtained. Among them, NN23 contained 25 amino acid substitutions with the activity improving 120%, whereas NN20 contained 24 amino acid substitutions with the activity improving 102%. EC1 contained 2 amino acid substitutions with the activity improving 47%. The rest 6 mutants contained one amino acid substitution with their activity increasing from 9% to 60%. The enzymatic characterization showed that all the evolved enzymes increased their catalytic efficiencies towards dichloroacetic acid and binding affinity towards glutathione whereas some of them increased the renaturability. However there is no obvious change in their thermostability. Based on these data, functional residues related to catalysis and refolding of AtGSTZ were discussed.]]> 2007/8/21 0:00:00 TAO Su-Dan,CHEN Xi-Wen,LIU Jia,JIA Xiang-Dong and CHEN De-Fu 950true -/-) mice in normal chow diet. The level of plasma lipids was also analyzed. Combined with the pathomorphological characteristics of the aortic root, the sequential expression characteristics of atherosclerosis related genes in the early stage of the atherosclerosis were studied. ApoE-/- mice compared with WT mice, the genes detected in the aorta, the expressions of VCAM-1, IκB and TGF-β in 1-month old of the apoE-/- mice were prominently up-regulated; while the expression of PDGF-α and CD36 were significantly up-regulated in 2-month old, the mRNA levels of TNF-α and MMP2 in 3-month old mice were prominently up-regulated, and IL-1β was significantly up-regulated from 1 to 3-month old. The genes detected in the liver, the mRNA level of CRP was significantly up-regulated at 3-month old of the apoE-/- mice. The mRNA level of NF-κB had no significant change at different ages in the aorta and liver in the two types of mice. Serum TC, TG and LDL-C levels in different ages of apoE-/- mice were significantly higher than those of WT mice. It appeared lipid deposition in the intima of aortic root in 2-month old apoE-/- mice. With the increasing of age, the lesions became more serious. The results implied that the sequential and differential expression of genes related to atherosclerosis formed a complex regulatory network which NF-κB is the core, and the genes may interact with each other and play important roles in the early stages of atherosclerosis of young apoE-/- mice.]]> 2007/9/5 0:00:00 JING Wen,YIN Miao,DU Hui-Qin,YE Hong-Yan,ZHANG Sheng-Qing,SHANG Yun-Ju,DAI Xue-Dong,QU Zhi-Ping,ZHANG Liang and PAN Jie 949true cis element of AAV genome essential for rAAV rescue, replication and packaging. It is prone to mutation or loss when it is latent in host cell or in plasmid. Plasmids with different ITR types were cloned to compare the influence of ITR types on the AAV packaging and infectivity. The vector plasmids were transformed the competent SURE cells to get different colonies. The ITR types of plasmids were screened by digestion with SmaⅠ. AAV vector plasmid pScGFPud has two ITRs at both ends of AAV genome and plasmid pScGFPu has only one ITR at upstream end of AAV genome. When the two plasmids were co-transfected 293 cells to prepare rAAVs, 1.08×1013 viral particles (AAV1-GFPud) were produced from 20-dishes of 293 cells cotransfected with plasmid pScGFPud, 4.28×1012 viral particles (AAV1-GFPu) were produced from 20-dishes of 293 cells cotransfected with plasmid pScGFPu. Virus AAV1-GFPud infected 293, HeLa and NCI H446 cells more efficiently than did virus AAV1-GFPu. This suggests that defect ITRs in AAV genome is deleterious to AAV packaging and AAV infectivity and vector with complete ITRs is favorable to the yield and activity of rAAV.]]> 2008/2/3 0:00:00 CAO Zuo-Wu,LIN Yi,CHENG Long-Qiu and ZOU Fei-Yan 948true 2007/8/15 0:00:00 CHEN Lan,CAI Lun,Geir Skogerbø and CHEN Run-Sheng 947true fgf genes) was identified as a feedback antagonist of FGF signaling in zerbrafish, mouse and human. Sef has been reported to function in different ways, however the regulation of Sef stability remains unknown. The possible role of c-Cbl in the regulation of Sef protein degradation was investigated. Results from coimmunoprecipitation and immunostaining assays reveal that hSef colocalizes and interacts with c-Cbl. Data suggest that the interaction between hSef and c-Cbl results in the ubiquitination and subsequent degradation of the hSef protein. It was proposed that c-Cbl may serve as a modulator to regulate Sef protein stability during FGF signal transduction.]]> 2007/5/31 0:00:00 REN Yong-Ming,RONG Zhi-Li,LI Zhi-Yong,CHENG Long,LI Ying-Hua,WANG Yin-Yin,REN Fang-Li,David M.Irwin and CHANG Zhi-Jie 946true 2007/11/8 0:00:00 XU Li-Jun,CHANG Xiu-Chun,YAO Hang-Ping and WU Nan-Ping 945true 2007/11/29 0:00:00 CHENG Wen,CHEN Fan,YU Ping and ZHONG Guang-Ming 944true 1-B29)-AAK-(A1-A21) which named HMPIDesB30 (human mini-proinsulin des B30) was synthesized and inserted into the Escherichia coli-yeast shuttle vector pPIC9K. The constructed plasmid, HMPIDesB30/pPIC9K was transformed into the GS115 cells of the methylotrophic yeast, Pichia pastoris, using electrotransformation. A transformant with a high copy number of the gene integrated into the chromosome was obtained by YPD plates which contains increasing concentrations of G418. After fed-batch fermentation, the protein was purified by macroporous resin, ion-exchange chromatography and precipitated with 10～20 mmol/L Zn2+. The purity of HMPIDesB30 is 95%，the expression level reached 1.0 g/L, and the recovery rate of purification is about 60%. The purified protein was characterized by 16.5% Tricine SDS-PAGE, HPLC, and mass spectrometry, and the molecular mass of the expressed products is in accordance with the cumulated value. HMPIDesB30 can be secretorily expressed by Pichia pastrois. The composition of the fermentation medium, the optimal condition of the fermentation and an effective method to purify the expression products from the culture were established.]]> 2007/7/13 0:00:00 GAO Jian-Kun,CAI Shao-Xi,FAN Kai,FENG Qiang,CHEN Hai-Rong,ZHANG Yi,HU Wei and YANG Ying-Bin 943true 2007/9/12 0:00:00 LUO Juan,ZHANG Wei,YU Bing,HU Xiao-Xiang and LI Ning 942true Acidithiobacillus ferrooxidans is a chemolithotrophic microorganism capable of using ferrous ions and sulphides as energy sources. This microorganism has an important role in the bioleaching of minerals. During this process, the bacteria are normally subjected to several stressing conditions, such as temperature changes, lack of nutrients or pH changes, which may affect the efficiency of the bacterial action. SELDI is a recent technology that allows for high-throughput proteomics studies. The Protein Chip SELDI technology was used to generate comparative protein profiles of Acidithiobacillus ferrooxidans grown under phosphate starvation or normal condition additionally adding Fe2+ as energy resource. There were 13 significantly differential expressed protein's peaks found by using SELDI Protein Chip technologies, which made a solid foundation for further isolation these low molecular proteins by adopting technologies such as HPLC etc.]]> 2007/11/8 0:00:00 HE Zhi-Guo,ZHONG Hui,LI Qing-Hua,GU Guo-Hua,HU Yue-Hua and LI Gui-Yuan 941true 2007/7/5 0:00:00 CHEN Tong-Sheng,WANG Xiao-Ping,SUN Lei,WANG Hui-Ying and WANG Long-Xiang 940true E.coli and detected by SDS-PAGE. Through Glutathione-Sepharose 4B affinity chromatography, a single pure band of the MLCK fragment was obtained. The phosphorylated myosin ATPase activities of the MLCK fragment, as well as its proteolytic fragment HMM and S1 were measured with the EnzChek Phosphate Assay Kit. The MLCK fragment stimulated phosphorylated myosin ATPase activity (Vmax=(19.426±1.669)-fold, Km=(0.486±0.106)μmol/L). Similar stimulation figures were obtained by measuring the ATPase activity of phosphorylated HMM and S1. The data suggests that MLCK could activate phosphorylated myosin, HMM and S1. In addition, in vitro motility assays demonstrated that increasing amounts of the MLCK fragment increased actin-myosin interaction and sliding. Also, the velocity of actin filaments could be enhanced on a gizzard smooth muscle myosin surface with the MLCK fragment. It is conclude that the non-kinase C-terminal domain of MLCK is independent of the phosphorylating mode for activation of myosin.]]> 2007/8/28 0:00:00 LIANG Ming-Li,CUI Ying,LU Guang-Yan and GAO Ying 939true In situ hybridization method was used to detect the expression of CaMKⅡ mRNA in hippocampus. The results are as follows: (1) Escape latency. In the first two Morris water-maze tests, there was a decreasing trend of escape latency in three groups, and the escape latency of RS group was much longer than that of control group. After physical exercise,in the last Morris water-maze tests, the diference of escape latency in three groups is not significant. (2) Searching strategy. In the first Morris water-maze test, there was a decreasing trend of marginal strategy and an increasing trend of taxis strategy in three groups, but the frequency of marginal strategy was higher and the frequency of taxis strategy was lower in RS group than that in SS and control group in the third and fourth day（P < 0.01）. However, there were no significant differences among three groups in the second and third Morris water-maze test. (3) Memory test. In the three memory tests, as for the distance ratioin between the origin platform quadrant to total distance, the RS group was the worst. The frequency of taxis strategy was much lower in RS group than that in SS and control groups in the first to third day(P < 0.01). (4) In situ hybridization detection showed that the expression of CaMKⅡ mRNA in dentate gyrus and hilus was much lower in RS group than that in SS and control groups (P < 0.01). It was concluded that recurrent but not single prolonged seizures could cause long-term effects on learning and memory, which may be associated with the down-regulated expression of CaMKⅡ mRNA in hippocampus. Physical exercise improved the learning capacity of RS group but with no effect on memory capacity of RS group.]]> 2007/7/11 0:00:00 Ni Hong,JIANG Yu-Wu,TAO Lu-Yang,LOU Jiang-Yan,WANG Zhe-Dong and WU Xi-Ru 938true 2008/10/29 0:00:00 SHENG Zhi-Ling,QU Mei-Hua,HE Hai-Jin and HE Rong-Qiao 937true 2008/5/8 0:00:00 ZHANG Li,LI Qing-Zhang,CHEN Jian-Hui,GAO Xue-Jun and TIAN Lei 936true 2008/6/19 0:00:00 WANG Shao-Peng,MA Liang-Hui,LI Ji-Xian,PENG Zhi-Gang and CAO Jing 935true ex vivo H5N1 virus infection study, alveolus epithelium were found to be more susceptible to avian influenza than trachea and bronchus epithelial cells. These results suggest that the human respiratory tract, to some extent, is permissive for avian influenza viruses. The currently-observed limited human to human transmission of H5N1 virus may be associated with the different abundance of SAα2,3Gal linkages in human upper respiratory tract among individuals.]]> 2008/7/16 0:00:00 ZHANG Zeng-Feng,FAN Xiao-Hui,LI Kang-sheng,HUANG Kai,LUO Dian-zhong,FENG Zhen-Bo,WEI Min-Yi,GUAN Yi,CHEN Hong-Lin and ZHANG Jin-Xia 934true P < 0.01) and Ago4 (P < 0.05), but without apoptosis. It suggests that Argonaute proteins may function in the cell cycle progression through a possible pathway that does not require small RNAs. The mechanisms underlying this phenomenon are still a puzzle for us.]]> 2008/5/27 0:00:00 JIANG Lin,PAN Shu-Juan,GE Yu-Zhi and YIN James Q(YIN Qin-Wei) 933true INa), delayed rectifier potassium current (IK) and transient outward potassium current (IA) of hippocampal CA1 pyramidal neurons in rats. All of the experiments were performing on hippocampal slices in vitro. Sample entropy (SampEn) and detrended fluctuation analysis (DFA) were employed to measure the complexity and fractal property of spontaneous firing of neurons. It was found that the spontaneous firing rate was significantly decreased, the entropy of ISI sequence was obviously reduced, and the long-range temporal correlations of ISI sequence were weakened after performing 10 mmol/L urethane. Furthermore, INa was significantly inhibited, but IK and IA were not statistically changed for 10 mmol/L urethane. This suggests that the fractal properties and complexity of underlying dynamics of the system have been reduced by urethane. And the reduction of complexity may because of inhibition of INa. The results demonstrate that apparently random fluctuations in neuron spontaneous firing are dictated by a complex deterministic process that imparts “long-term” memory to the dynamic system.]]> 2008/8/20 0:00:00 LEI Ting,YAN Rui,LI Ya-Tang,YANG Zhuo and ZHANG Tao 932true 2008/6/19 0:00:00 REN Jian-Ke,SHEN Jia-Juan,SHENG Zhe-Jin and FEI Jian 931true in vitro differentiation.]]> 2008/7/16 0:00:00 ZENG Qiao,AN Shi-Min,PAN Qian,XIA Kun,XIA Jia-Hui,ZHANG Zhuo-Hua,SUN Yi and Fan Guo-Ping 930true IC50 was significantly increased in MDR cell line K562/Dox after PHT and CBZ was applied; verapamil could decrease significantly the level of IC50 in MDR cell line K562/Dox, and the reversal index were 2.5 and 1.5. The concentration of PHT and CBZ in MDR cell line K562/Dox was lower than that in non-MDR cell line K562, and verapamil significantly increased the concentration of PHT and CBZ in MDR cell line K562/Dox(P < 0.05). It is obvious that overexpression of PGP are concerned with drug resistance of medically intractable epilepsy.]]> 2008/8/19 0:00:00 CHEN Ying-Hui and ZHAO Yong-Bo 929true Drosophila models were constructed by using the promoter GMR-GAL4 and elav-GAL4 which drive target selective gene expression in developing eyes and neurons, respectively. Then, Hsp22 protein was overexpressed in SCA3/MJD transgenic Drosophila models at different levels by genetic methods and heat shock reaction. Overexpression of endogen Drosophila Hsp22 can notably suppress the neurotoxicity of MJDtr-Q78 protein, and the level of Hsp22 expression was in consistent with rehabilitation for neurodegeneration of Drosophila eyes, and extension of Drosophila lifespan. It is firstly confirmed that expression of Hsp22 protects the SCA3/MJD from neurodegeneration on Drosophila models, which might contribute to a potential therapeutic effect on SCA3/MJD.]]> 2008/10/13 0:00:00 LI Qing-Hua,JIANG Hong,YI Ji-Ping,LIAO Shu-Sheng,SHEN Lu,PAN Qian,XIA Kun and TANG Bei-Sha 928true Bt cry1Ac gene and bar gene expression cassettes, was constructed by using molecular method. The Bt gene expression cassette contained the 3.5 kb Bt cry1Ac gene under the control of psbA promoter and terminator cloned from sugar beet chloroplast genome. The bar gene expression cassette contained the bar gene, 16 S promoter and terminator cloned from sugar beet chloroplast genome, The atpB and rbcL gene cloned from sugar beet chloroplast genome were used as homologous fragment, the bar gene was the selective marker. Plasmid pSKARBt/bar were transformed into the petioles of sugar beet with particle bombardment method. The petioles were planced onto the shoot-inducing selection medium which contained spectinomycin (20 mg/L), 6-BA (1.5 mg/L) and NAA (0.2 mg/L) at first. And when the green shoots regenerated, the green shoots were transfered into the shoot-propagation medium for optimal shoot development which contained spectinomycin (20 mg/L) and 6-BA (0.5 mg/L) and NAA (1.0 mg/L) one subculture at 20-day intervals, and then the shoots were transfered into the shoot-propagation medium for optimal shoot development with herbicide (PPT 10 mg/L) several subcultures. The shoots were transfered into the root-induction medium with herbicide (PPT 10 mg/L) and the transgenic plants were obtained at last. The transgenic sugar beet plants were testsed by PCR and Southern blot. The results showed that the Bt gene and bar gene had been transferred into the chloroplast genome of sugar beet. The transgenic plants had tolerance to both PPT and bioassays testsed. The insecticidal activity (the mortality of larvaes was 33%～80%) and herbicide resistance of the transgenic plants indicated that the relevant protein had been expressed already in sugar beet. The study showed that the bar gene can also be used as an efficient selective marker gene besides antibiotic resistant markers in plant transformation. Efficient transformation system in sugar beet chloroplast had been established.]]> 2008/11/3 0:00:00 CUI Jie,LI Bin-Sheng,YANG Qian and CHENG Da-You 927true 2+ sensor of calcium release-activated calcium (CRAC) channel that is constructed by membrane protein Orai1. However, this regulatory system may also be regulated by other proteins. Reticulocalbin 2 (RCN2) was purified and identified from STIM1-Orai1 complex. Confocal microscopy revealed that RCN2 co-localized with STIM1 in ER before and after Ca2+ store depletion. Single cell [Ca2+]i measurements of RCN2 EF hands mutant showed slight influence on SOC electrophysiological characters. Furthermore, a novel collar form aggregation of RCN2 surrounding STIM1 clusters suggested that RCN2 potentially plays a role of structure maintenance in STIM1 clustering.]]> 2008/9/16 0:00:00 ZHAN Yi,GAO Shang-Bang,XUE Peng,YANG Xiao-Fei,LI Zheng-Zheng and XU Tao 926true 2008/10/29 0:00:00 HU Gang-Qing,LIU Yong-Chu,ZHENG Xiao-Bin,YANG Yi-Fan,SHE Zhen-Su and ZHU Huai-Qiu 925true via Skp2 in SMMC-7721, however which was Akt independent.]]> 2008/5/8 0:00:00 FU Yi,FANG Zheng-Yu,WANG Li-Ying,LI Zeng-Xia,YANG Yong and ZHA Xi-Liang 924true 2 to hydrogen peroxide treatment, and to search new targets for cancer chemotherapy, HepG2 cells were transfected using lipofectamine 2000, and selected by treatment with G418. Stable cell lines were confirmed by reverse transcriptase polymerase chain reaction (RT-PCR) targeting vector gene. Experiments include the following groups: (1) control group (non-transfected cells); (2) transfected with empty vector (HepG2/GFP cells); and (3) transfected with pEGFP-C1-Daxx (HepG2/GFP-Daxx cells). After incubation with hydrogen peroxide (H2O2) for 24 h, cellular viability was analyzed by MTT, and cellular apoptosis was measured by flow cytometric analysis. Gene expression at protein level was detected by Western blot. The RT-PCR results showed that Daxx RNA in cells transfected with pEGFP-C1-Daxx was increased significantly compared with that in the HepG2/GFP cells. Fluorescence microscopy revealed that Daxx protein was localized in the nuclei. Hydrogen peroxide was used to induce apoptosis of HepG2 cells and observed that the hydrogen peroxide decreased the viability of HepG2 cells in concentration-dependent pattern. The IC50 values in three groups (Normal cells, HepG2/GFP cells and HepG2/GFP-Daxx cells) were 0.72, 0.76, and 0.49 mmol/L respectively. The apoptotic ratio was significantly higher in HepG2/GFP-Daxx cells as compared to the other two groups. HepG2/GFP-Daxx cell incubated with hydrogen peroxide, showed a significant increase in the activation of caspase-3 and JNK as compare with the other groups. Over-expression of Daxx facilitated HepG2 cells apoptosis induced by hydrogen peroxide. Furthermore, there may be a synergetic relation with apoptosis and increase of JNK activity.]]> 2008/6/19 0:00:00 TUO Qin-Hui,XIONG Guo-Zuo,ZHU Bing-Yang,CAO Jian-Guo and LIAO Duan-Fang 923true FDDV in immune suppressed horses were detected. Three horses, which were immunized with EIAVFDDV for 16 months, were treated with dexamethasone for 14 days to suppress their immunities. Reduced immune response in these animals was confirmed by significantly declined lymphocyte proliferation rate detected after 10 days of the drug treatment. The plasma viral loads of EIAVFDDV, which was indicated by the genomic RNA copy numbers, in horses before and after the treatment of dexamethasone were monitored by real time RT-PCR. Results revealed that the viral plasma loads in two of three immune-suppressed horses were kept a steady low level around 103～104 copies/ml. The load was increased by 10 folds in the third horse, but was still among the standard levels for EIAVFDDV vaccinated horses. As a positive control, the viral copy number of an asymptomatic carrier of EIAV virulent strain EIAVLiao was jumped nearly 25 000-fold higher after being treated with dexamethasone. The typical clinical symptoms of EIA, characterized by febrile episodes and thrombocytopenia, were also appeared in this horse. These results clearly indicate that it is the unique biological feature of the attenuated EIAV vaccine, but not the immunity, resulted in EIAVFDDV remaining in low levels in the body harmlessly. In addition, the steady low level of viremia and the inability to cause clinical symptoms of EIAVFDDV in immune-suppressed hosts further demonstrated the safety of attenuated Chinese EIAV vaccines. The data provide a new sight for studies on the immunity to lentiviruses.]]> 2008/5/8 0:00:00 MA Jian,JIANG Cheng-Gang,LIN Yue-Zhi,GUO Liang,GUO Wei,KONG Xian-Gang,SHEN Rong-Xian,SHAO Yi-Ming,ZHANG Xiao-Yan and ZHOU Jian-Hua 922true beclin 1 regulates cell growth through autophagy. The mRNA expression of beclin 1 was reported to be down-regulated in breast cancer with high frequency of loss of heterozygosity (LOH). However, there was no report about the expression levels or the regulatory mechanisms of beclin 1 in gastric and colorectal cancer. Both the mRNA and protein expression levels of beclin 1 was detected in the tissues of gastric and colorectal cancers, as well as the aberrant DNA methylation and LOH related to the expression of beclin 1. By comparing with normal tissues adjacent to the tissue of these tumors, it was found that beclin 1 mRNA expression levels were significantly decreased in gastric tumor tissue. Furtherly by explorating the 5′ region of beclin 1 gene sequence, a large and dense CpG island was discovered and meanwhile methylations in the promoter and the intron 2 regions of beclin 1 were found in both gastric and colorectal tumors. And LOH was found in gastric tumors. These findings suggested that aberrant DNA methylation, as well as LOH, were involved in the regulation of beclin 1 expression in gastric and colorectal cancer.]]> 2008/6/19 0:00:00 LI Zi-Dong,CHEN Bo,WU Yi-Qing,JIN Feng,XIA Yong-Jing and LIU Xiang-Jun 921true P < 5%) were identified: 9 on chromosome 2 affecting WBC, GRAN, MCV, HGB, MCHC, PLT, MPV, PDW and PCT, respectively; 7 on chromosome 7 affecting WBC, GRAN, MCHC, HGB, PLT, MCV and RDW, respectively; 6 on chromosome 8 affecting GR%, LY%, MCHC, PLT, PCT and MCV, respectively. In order to avoid the increase of false positive rate caused by multiple tests, the FDR (false discovery rate) control approach was adopted to further test these 22 QTL. The results indicated that 14 of them were significant at FDR < 5% level, of which 9 were significant at FDR<1% level.]]> 2008/8/19 0:00:00 GONG Yuan-Fang,LU Xin,WANG Zhi-Peng,LI Shan,QIU Xiao-Tian and ZHANG Qin 920true Caenorhabditis elegans (C. elegans) in spaceflight was carried out, which had been reported in this journal before. An environment of simulated microgravity on ground was established, and its major effects on body-wall muscles of C. elegans in the structures and functions were examined, which further confirmed the results from spaceflight studies, and comparing between these two different treatments was benefit for valuing the validity of simulated microgravity. Firstly, the survival rate and movement ability of C. elegans were observed, and five important muscle-related genes and three proteins were measured after 14 days 19.5 h rotation. The animals displayed reduced rates of movement with a lower ratio (height/width) in crawl trace wave in simulated microgravity, indicating a functional defect. In morphological observation deceased muscle fiber size in myosin immunofluorescence and duller dense-body staining were found in microgravity group, suggesting muscular atrophy had happened in C. elegans. Meantime the result of Western blotting showed the quantity of myosin A decreased significantly in simulated microgravity group, further confirming muscular atrophy. In genes transcription, it was noted that dys-1 increased significantly in body-wall muscles, while hlh-1, unc-54, myo-3 and egl-19 mRNA levels declined after rotation. This study provided evidence that dys-1 are involved in the transduction of mechanical information in skeletal muscle, potentially play a vital role in gravisensing. Genes of hlh-1, unc-54, myo-3 and egl-19 induced the muscular atrophy in simulated microgravity from the structures and functions ways respectively. Data of this study consolidated the results in our spaceflight researches. On the other hand, it is implied that simulated microgravity is an effective ways for improving the quality of space studies.]]> 2008/7/9 0:00:00 WANG Chi,SANG Chen,YANG Chun,SUN Yan,YI Zong-Chun and ZHUANG Feng-Yuan 919true Caenorhabditis elegans gene FAT-2 encoding △12 fatty acid desaturase in CHO cells was reported. First, the FAT-2 coding sequence was cloned by RT-PCR. To facilitate high level synthesis of heterogeneous protein, the codon usage of the fatty acid desaturase genes was optimized according to the codon preference of mouse by site-directed mutagenesis, 2 synonymous mutations were introduced into FAT-2 gene by overlapping PCR. The codon-modified gene was finally fused to pBudCE4.1 vector (Invitrogen) under the control of CMV promoter. The expression vector pBudCE-FAT2 was linearized with NheⅠ, and then transfected CHO cells, the cells were under Zeocin selection for nine days and then propagated, then the transfected cells were harvested. The genome and total RNA were isolated for PCR and Norhern blot ananlysis. The results revealed that FAT-2 gene has been integrated into the genome of CHO cells and expressed properly. Fatty acids of total cellular lipids were analyzed by gas chromatography. The results indicate that the expression and function of △-12 fatty acid desaturase resulted in accumulation of linoleic acid. The levels of linoleic acid in transgenic cells were 2.4-fold higher than those in wild-type cells. The moderate linoleic acid in CHO cells was derived from cell culture media uptaken by cell membrane. The results demonstrate that a heterogenous desaturase gene can function well in mammalian cells and prove that transgenic approach is an efficient strategy for changing fatty acid composition of mammals.]]> 2008/6/19 0:00:00 KONG Ping,DU Zhuo,TANG Bo,MENG Qing-Yong and LI Ning 918true S. typhi, and dynamics of MAP/mmHg in mice injected with LPS were observed. The results showed that specific antibodies against S. typhi-LPS 7261 and E. coli-LPS 2630 were elicited in mice immunized with peptide 13L-BC conjugate, and could be boosted by inactive bacterial and LPS. The main isotype of anti-LPS antibodies were IgG2a, IgG2b and IgM, but less IgG1 and IgG3, The survival time of mice infected with S. typhi were (12±1.3) days and (5.3± 0.4) days(P < 0.01) in group immunized with peptide 13L conjugate and with BC, respectively. And the peptide 13L-BC can also elicit protective immunity against endotoxic shock by LPS. The dynamics of MAP/mmHg observed by carotid Artery catheterization showed a significant difference between mice immunized with peptide 13L-BC and mice immunized with BC only in 1, 2,3 and 4 hours after injection of LPS 2630(P < 0.05, P < 0.01, P < 0.05 and P < 0.01), and injection of LPS 7261(P > 0.05, P < 0.05, P < 0.05 and P < 0.01). These results suggested that the peptide 13L can mimic the antigenicity of LPS epitopes to induce secondary antibody response like as thymus-dependent antigen, and also can elicit a protective immunity of mice from infection with G- bacteria and endotoxic shock. This peptide mimics could be a new vaccine candidate of LPS.]]> 2008/7/16 0:00:00 LUO Hai-Bo,ZHENG Jian,ZHU Ping,HUANG Lai-Qiang and FU Ning 917true Oryza sativa mitochondria tRNATrp toward B． subtilis tRNATrp were constructed and transcribed in vitro with T7 RNA polymerase. The kinetic parameters (Kcat/KM) of B． subtilis tryptophanyl-tRNA synthetase(TrpRS) and human TrpRS were determined with four mutant-type tRNATrps. Results showed that for reaction with B． subtilis TrpRS, C2/G71 and C4/G69 mutations each induced a comparable 40-fold and 53- fold of activity to Oryza sativa mitochondria tRNATrp respectively. Notably, when the C2/G71 and C4/G69 mutations were introduced together into B． subtilis tRNATrp, a 140-fold of reaction rate resulted, the catalytic efficiency was 34 percent as that of wild-type B． subtilis tRNATrp, but these four mutants resulted in a weak aminoacylation efficiency by human TrpRS, and the change was little. Clearly, the results indicate that C2/G71 and C4/G69 bases in the acceptor stem are important species-specific elements of Oryza sativa mitochondria tRNATrp, since which are significant to the aminoacryl activity.]]> 2008/6/10 0:00:00 JIN Xiao-Ling,GONG Ju-Fang,LIU Xue-Mei,WANG Xiao-Hong and ZHANG Ri-Qing 916true 1H NMR spectroscopy in conjunction with principal component analysis (PCA). With rich metabolite information, 1H NMR spectra showed that the tumor-tissue metabonome was dominated by lipids, lactate, myo-inositol, creatine, choline metabolites such as choline, phosphocholine and glycerophosphocholine, amino acids such as alanine, glutamate, glutamine, taurine, N-acetyl-aspartate and glutathione. PCA of the tumor NMR spectra clearly showed metabonomic differences between low-grade astrocytomas and meningiomas whereas such differences were more moderate between fibroblastic and transitional meningiomas. Compared with meningiomas, the low-grade astrocytomas had higher levels of glycerophosphocholine, phosphocholine, myo-inositol and creatine but lower levels of alanine, glutamate, glutamine, glutathione and taurine. The N-acetyl-aspartate level was low but detectable in low-grade astrocytomas whereas it was not detectable in meningiomas. It is concluded that tissue metabonomics technology consisting of HRMAS 1H NMR spectroscopy and multivariate data analysis (MVDA) offers a useful tool (1) for distinguishing different types of brain tumors, (2) for providing the metabolic information for human brain tumors, which are potentially useful for understanding biochemistry of tumor progression.]]> 2008/5/29 0:00:00 CHEN Wen-Xue,LOU Hai-Yan,ZHANG Hong-Ping,NIE Xiu,XIANG Yun,YANG Yong-Xia,WU Guang-Yao,QI Jian-Ping,YUE Yong,LEI Hao,TANG Hui-Ru and DENG Feng 915true et al), the changes of gene/protein expression profiles and the down-regulation of expression of phosphor-EGFR. Based on these studies, in an attempt to identify the function of NGX6-induced-apoptosis, the following experiments were designed. The NGX6-transfected HT-29 cell line was used as the test, empty-vector- transfected HT-29 cell line and untransfected HT-29 cell line were used as the control. The effect of NGX6 on apoptosis was detected by FCM cells were double- stained by PI/Annexin-V; the express of NF-κB was detected by EMSA. There is no difference of apoptosis between NGX6 transfected colon carcinoma cell and NGX6 untransfected colon carcinoma cell when cells are cultivated in vitro. But apoptosis level of NGX6 transfected colon carcinoma cell of xenograft tumor in nude mice is higher than that of NGX6 untransfected colon carcinoma cell. And the express of NF-κB is inhibited in NGX6 transfected colon carcinoma cell groups. These experiments showed NGX6 gene can induce apoptosis of colon cancer and inhibit The express of NF-κB.]]> 2008/8/19 0:00:00 LIAN Ping,GUO Qin,PENG Ya,XIAO Zhi-Ming,LIU Fen,WANG Xiao-Yan,SHEN Shou-Rong and LI Gui-Yuan 914true in vitro. The experimental results showed that the 5(6)-TAMRASE can conjugate with YSL stably. When incubated with human hepatocellular carcinoma BEL-7402 cells, 5(6)-TAMRASE labeled YSL could entered the cells in 15 min, and mainly concentrated in the cytoplasm of BEL-7402 cells. Then the fluorescent intensity of 5(6)-TAMRASE labeled YSL which indicated the concentration of them reach the maximum until incubated with the cells for 2 h. After that it began to go down. The 5(6)-TAMRASE could also enter the hepatocellular carcinoma BEL-7402 cells, but it distributed in the whole cell and the intensities had no decrease within 3 h. The results suggest that the distribution pattern of fluorescent labeling YSL is different from that of 5(6)-TAMRASE. The subcellular distribution pattern of 5(6)-TAMRASE labeled YSL was ascribed to YSL and could represent that of YSL. In conclusion, YSL located at the cytoplasm of hepatocellular carcinoma BEL-7402 cells and distributed intensively.]]> 2008/6/19 0:00:00 JIAN Xu,FU Zheng,ZHANG Yan-Ling,HAN Li-Xia,ZHU Zhi-Feng,XU Qiong,WANG Li,ZHOU Chun-Lei,SHI Lin-Xi,LIU Jun-Yan,LU Rong and YAO Zhi 913true A. oxyphylla on the proliferation capacity of hADSCs in vitro was examined. Human ADSCs could differentiate into neuron-like cells in vitro, and protect PC-12 cells from apoptosis induced by serum deprivation. Cell counts showed that treatment of hADSCs with 0.5 mmol/L, 1.0 mmol/L and 1.5 mmol/L of PCA for 48 h increased the cell numbers in a dose-dependent manner. In addition, the cell numbers of hADSCs at various time points after treatment of 1.5 mmol/L PCA were increased in a time-dependent manner. Flow cytometric analysis of DNA content demonstrated the cell cycle progress from G1 phase to S phase. The most pronounced effect was seen with 1.5 mmol/L PCA, where the fraction of cells in S phase increased more than 2 folds, accompanied by a significant increase in the fraction of cells in G2/M phase and a significant decrease in the fraction of cells in G0/G1 phase. Western blot analysis revealed the elevated expression of cyclin D1 in hADSCs induced by 1.5 mmol/L PCA treatment. Furthermore, cyclin D1-siRNA transfection significantly inhibited the promotion of cell proliferation by PCA. Flow cytometric analysis of the cell surface antigens, osteogenic induction and adipogenic induction demonstrated that after PCA treatment, hADSCs retained their morphological and functional characteristics of multipotential mesenchymal progenitors. The proliferative enhancement of PCA suggests the possibility that PCA may be useful in hADSCs-mediated therapy.]]> 2008/10/23 0:00:00 WANG Han,LIU Tian-Qing,ZHU Yan-Xia,GUAN Shui,MA Xue-Hu and CUI Zhan-Feng 912true IR) of the three cell lines which were affected by carboplatin in different concentrations. Accordingly, the IC50 of the three cell lines could be calculated. Carboplatin induced apoptosis of three cell lines was observed by flow cytometry (FCM). In addition, the apoptotic ratios of the three lines that were treated by 30 mg/L and 60 mg/L carboplatin were measured by the FCM at the same time. And then, the apoptosis conditions of the three lines were observed by the methods of fluorescent staining and transmission electron microscope (TEM). The IC50 of RMG-Ⅰ-H was (58.07±2.42), which was obviously higher than that of RMG-Ⅰ(28.83±3.57) and RMG-Ⅰ-C(25.71±8.24), all showed a significant statistical difference (P < 0.01) and there was no statistical difference between the latter two ones(P > 0.05). FCM analysis confirmed the cell apoptosis in the three lines. The corr-apoptotic ratios of RMG-Ⅰ-H treated with 30 mg/L and 60 mg/L carboplatin were (20.43±0.71)% and (38.11±0.33)%, respectively, which were about 49%～63% of the RMG-Ⅰ(35.87±3.84)%, (63.37±9.59)% and RMG-Ⅰ-C(34.80±3.59)%, (60.17±6.64)%, both presented a statistical difference (P < 0.05). But there was no significant statistical difference between the latter two ones (P > 0.05). The apoptotic degree of RMG-Ⅰ-H was lower than that of the other cell lines in the same carboplatin concentration with the method of inv-fluorescence microscopy and transmission electron microscopy (TEM). The drug resistance of ovarian cancer cells to carboplatin was stronger when the Lewis(y) antigenic contents on cell surface increased.]]> 2008/3/25 0:00:00 ZHAO Yue,LIN Bei,HAO Ying-Ying,YAN Li-Mei,LIU Juan-Juan,ZHU Lian-Cheng and ZHANG Shu-Lan 911true GTL2 is imprinted in human and mouse, which act as noncoding RNAs regulating the translation of target mRNA. In order to identify the expression of GTL2 in cattle produced by natural reproduction and by SCNT, a single nucleotide polymorphisms (SNPs) in GTL2 through PCR-SSCP was identified and the GTL2 expression patterns in six organs of hybrids was analyzed by RT-PCR-SSCP. The results demonstrated that GTL2 was monoallelic expression in all six examined organs of cattle produced by natural reproduction, and showed monoallelic expression in heart and liver but biallelic expression in brain, spleen, lung and kidney of cattle produced by SCNT that died shortly after birth. The abnormal expression of GTL2 may contribute to the organ development defects and the low efficiency of SCNT.]]> 2008/9/2 0:00:00 WANG Iian-Chun,LI Dong-Jie,DU Wei-Hua,FAN Bao-Liang,LI Ning and lI Shi-Jie 910true 2008/5/8 0:00:00 ZHANG Li-Rong,LUO Liao-Fu,XING Yong-Qiang and JIN Hong-Ying 909true Caenorhabditis elegans (C. elegans) in the structures and functions were examined, and five important muscle-related genes and three proteins were studied after nearly 15-day spaceflight. The changes for the wall-muscles were observed in situ. Decreased muscle fiber size was observed with myosin immunofluorescence and duller dense-body staining in flight samples, which suggested that muscular atrophy had happened during spaceflight. However, F-actin staining showed no differences between the spaceflight group and ground control group. Otherwise, after returning to the earth the C. elegans displayed reduced rate of movement with a lower ratio (height/width) in crawl trace wave, which indicated a functional defect. These results demonstrated that C. elegans muscular development was changed in response to microgravity, and changes also occurred at the level of gene transcription and protein translation. Expression of dys-1 increased significantly in body-wall muscles, while hlh-1, myo-3, unc-54 and egl-19 RNA levels decreased after spaceflight. Dystrophin (encoded by dys-1) is one of important components in dystrophin-glycoprotein complex (DGC). Increased dys-1 expression after flight implied that the muscular cell would accept more gravity signals by DGC in microgravity in order to keep mechanical balance within the cells. It is concluded that DGC was involved into the mechanical transduction in body-wall muscles of C. elegans when gravity varied, which potentially played a vital role in gravisensing. The changes of hlh-1, myo-3, unc-54 and egl-19 suggested that they had the effects of promoting microgravity-induced muscular atrophy in structure and function aspects. Result of Western blotting showed that the level of myosin A in spaceflight group decreased, further confirmed that atrophy happened during flight.]]> 2008/10/23 0:00:00 WANG Chi,SANG Chen,Akira Higashibata,Noriaki Ishioka,RONG Long,YANG Chun,SUN Yan,YI Zong-Chun and ZHUANG Feng-Yuan 908true 2008/9/16 0:00:00 SHENG Zhi-Ling,LIU Yan-Ying,CHENG Lan and HE Rong-Qiao 907true 2009/3/30 0:00:00 CAO Qiu-Ju,TIAN Zhi-Hua,SUN Sheng,YANG Ning,WANG Fang,HUANG Ling-Yun,PENG An,LIU Hui-Tu and ZHANG Wei 906true 2009/3/26 0:00:00 LI Mei-Xiang,XIAO Zhi-Qiang,PENG Fang,LI Guo-Qing,ZHANG Peng-Fei,LI Mao-Yu,LI Cui,LI Feng,LIU Ying-Fu and CHEN Zhu-Chu 905true 2009/5/14 0:00:00 WANG Hai-Zhen,WANG Ying,SUN Da-Quan and LIU Ding-Gan 904true 2009/5/13 0:00:00 AN Qun-Xing,LEI Ying-Feng,YANG Jing,ZHANG Xian-Qing,YI Jing,CHEN Rui and MU Shi-Jie 903true C) were expressed and purified, and their solubility and activity were compared. The experiment results show that the solubility of F185K mutated INC was dramatically increased, whereas the activity was reduced to some extent. Subsequently, 1 800 ps molecular dynamics (MD) simulations for the WT and F185K type of INC in water were performed. The MD simulation results demonstrate that the flexibility of the catalytic loop region and the total mobility of F185K INC was reduced, which causes the decrease of activity. After the F185K mutation, changes of the salt bridge network drove the conformational change of INC, resulted in the burying of some hydrophobic residues and exposure of some other hydrophilic residues on the protein surface. Therefore, the relative hydrophilic solvent accessible surface of INC was increased. Moreover, the F185K mutation increased the hydrogen number between the INC protein and water molecule, as a consequence, the protein-water interaction was enhanced. These above changes contribute to the solubility increase of INC. It is found that the results obtained from MD simulation are in good agreement with the experiment data. The above mentioned results provides valuable insight for the understanding of protein solubility and will be helpful in protein engineering for increasing the solubility of proteins.]]> 2009/5/18 0:00:00 HE Hong-Qiu,HU Jian-Ping,LIU Bin,CHEN Wei-Zu and WANG Cun-Xin 902true 2009/6/8 0:00:00 GUO Zhi-Yun,MAO Can-Quan,XIONG Li-Li and XIN Hong-Bo 901true -/- mice was investigated. Fifty male apoE-/- mice (5wk old) were divided into four groups: basic diet group (control group), high-fat diet group (control group), EGCG+ basic diet group (EGCG group), and high-fat diet + EGCG group (EGCG group). EGCG (10 mg/kg) was injected intraperitoneally every day. Areas of aortic plaque areas were measured by oil red O staining. Western blot was used to detect the expression of TLR4 and CD14 in mouse aorta. The expression of TLR4 mRNA and CD14 mRNA were detected by Real-time PCR. Serum concentrations of MCP-1 and TNF-α were determined with ELISA. Compared with high-fat diet group, EGCG groups showed marked decreases in aortic atherosclerosis(P < 0.05), concomitantly with significant decreases in levels of the expression of TLR4, TNF-α and MCP-1. The mean lesion area was (2.37 ± 0.08) mm2 in the high-fat diet + EGCG group, whereas the atherosclerotic lesion was only (1.05 ± 0.13) mm2 in the high-fat diet + EGCG group. The TLR4 expression was obviously higher in high-fat diet group than that in other groups (P < 0.05). Compared with high-diet group, the serum concentrations of MCP-1 and TNF-α were significantly decreased in EGCG groups (P < 0.05). These results suggest that TLR4 signaling pathway may play an important role in the development of atherosclerosis in apoE-/- mice induced by high-fat diet.]]> 2009/7/9 0:00:00 GU Hong-Feng,ZHOU Hao,TANG Chao-Ke,SUN Hui and YANG Yong-Zong 900true 2009/5/27 0:00:00 LIU Lei,ZHANG Ying-Jie and WANG Xian-Wang 899true 2009/5/13 0:00:00 ZHANG Ying,SHI Fang-Yu,GAO Shao-Rong and LIN Ai-Xing 898true 2009/4/24 0:00:00 ZHANG Chuan-Shan,GUO Yi,GU Rui-Huan,LI Shan-Gang,LI Feng,WANG Wei,DING Lei,XING Feng-Ying,YAO Gang and CHEN Xue-Jin 897true in vivo and vitro. The results showed that Tb4 were different in expression and localization during mouse oocytes maturation and early embryonic development. Immuno-histochemistry results illustrated that, accompany with follicle growing, the expression amount of Tb4 peptide in vitro oocytes is gradually increased. There are different distribution characteristics of Tb4 in GV stage oocytes, which featured by chromatin configuration, such as divided the configuration into NSN, pNSN and SN type. In other stages of oocytes maturation and early embryonic development, Tb4 could locate both in nuclear and cytoplasm, and the expression amount always changing regularly, which are closely related with cells growing status and the cell cycle. AIOD value of Tb4 was not significantly different in the process of oocytes maturation in vivo, but the fluorescence intensity rapidly increased form 8-cell embryos to blastula stages in the process of embryonic development. The tendency of Tb4 mRNA transcription are similar to the peptide expression in those stages. Specifically, in early implantation blastula, Tb4 peptide highly expresses in implanted lateral cells of blastula, indicating that Tb4 can regulate the microfilaments depolymerization in embryos implantation, so Tb4 plays an important role in the process of embryos implantation. The research concluded that, Tb4 could regulate microfilaments events (such as polymerization and depolymerization) and proliferation of cells in the developmental processes through the changes of its expression and localization. Overall, Tb4 plays an important role for mouse oocytes maturation, early embryonic development and embryos implantation.]]> 2009/3/30 0:00:00 ZHANG Yan,LI Shu-Feng,YANG Cai-Rong,ZHENG Ke-Jia,LI Ning and YAN Yun-Qin 896true tak1 and tak2 influenced by the change of light and temperature in RT-PCR with specific primers and the possible regulation relationship between TAK1 and LHCⅡ phosphorylation especially by using TAK1 serum and P-Thr antibody. The result indicated that light and temperature regulated the phosphorylation of LHCⅡ and took effect on the transcription and translation level of tak1 and tak2, but the response of LHCⅡ phosphorylation to light did not coincided with the quantity of TAK1 protein. Furthermore, low light density could enhance the expression of tak2, but temperature impacted tak2 little. The different regulation pattern of tak1 and tak2 may derive from the different elements of promoter.]]> 2009/5/25 0:00:00 JIANG Jia-Hong,WANG Dong,HU Yuan,GAO Xiu and DU Lin-Fang 895true E. coli. system. Polyclonal antibodies were generated by rabbit immunization. Western blotting analysis were carried out for rice material collected at different developmental stages including shoot and root at seedling stage, root and stem at tilling stage, flag leaf and young panicle at heading stage, flag leaf and panicle at flowering stage, flag leaf and seed at filling stage. Comparison analysis was carried out with EST sequencing data. Specific antibodies were obtained by rabbit immunization of recombinant proteins expressed in E. coli. One major band were observed for Western blotting detection of rice U-Box proteins, the apparent molecular mass of two U-Box proteins (Os06g01304 and Os12g38210) were consistent with predicted size, while the other two proteins (Os01g66130 and Os08g01900) with apparent molecular mass smaller than that of predicted. Western blotting results indicated that the U-Box proteins were constitutively expressed with close abundance in tested tissues. EST analysis based on 1 million ESTs derived from 274 libraries from NCBI EST database revealed closed numbers of U-Box gene transcription, which is parallel with the results of constitutively expression of proteins. However, compared with the expression of ATPase, HSP81-3, EGF-1 alpha and RuBisCo, the number of ESTs for U-Box proteins were much lower, suggest the low abundance transcription of U-Box genes. Four rice U-Box proteins were chosen for the generation of specific antibodies via the expression of predicted epitope fragments, demonstrated the feasibility of the process. Western blotting analysis indicated that four U-Box proteins were constitutively expressed among tested rice tissues at different developmental stages, which is parallel with EST sequencing data. The antibodies will provide resources for functional studies, such as co-immunoprecipitation, ChIP-on-chip, Pull-down and stress response etc.]]> 2009/4/15 0:00:00 CHEN Hao,LI Li-Yun,BAI Hui,CAO Ying-Hao,WANG Xian-Yun and LIU Guo-Zhen 894true CYP72B1 and AUR3 are two important genes responsive to light, auxin and brassinosteroid at transcription level. To understand the regulation mechanism of the two genes, a new tool called OCMMat was developed for identifying cis-elements, OCMMat combines both the over-representation property of regulatory elements in co-expressed genes and the conservation property in orthologous genes, for the latter, it was estimated by an enrichment score of regulatory element in orthologous promoter sequences. Using this tool, 3 regulatory motifs shared by genes CYP72B1 and AUR3 were reported, motif GAGACA which is the same as a known cis-element AuxRE, motif AAGAAAAA containing the sequence of GT element and the third ATCATG which is a new one named EDIB element. The space and the order of AAGAAAAA and EDIB show the same pattern in promoters of both the co-expressed genes and the orthologs of CYP72B1. Based on the sequence analysis and the literature knowledge to date, a model was proposed for describing the transcriptional regulatory mechanism of CYP72B1 and AUR3 in response to light, auxin and brassinosteroid. The model presents how the signaling pathways of light, auxin and brassinosteroid are interplaying at gene transcription level. In response to light, the transcription factors GT factor and an unknown protein repress the expression of CYP72B1 and AUR3, the hormone pathways are not interfered and thus work in their own way. While in the absence of light stimulation, CYP72B1 and AUR3 are expressed and the products, in turn, inhibit both the auxin pathway and the brassinosteroid pathways. On the other side, at high hormone level, gene expression is up-regulated through ARF binding, the gene products inhibit the hormone pathways in a feedback manner, and meanwhile, rescues the light signal through the photoreceptor phyB.]]> 2009/7/9 0:00:00 ZHANG　Chang-Qing,WANG Jin,ZHU Huai-Qiu and GAO Xiang 893true Pichia pastoris, and purified by chromatography. EC1.71 cells expressed high levels of NGALR (NGAL receptor) and EC109 cells expressed low levels of NGALR were used as cells model. The trafficking and the possible function of NGAL protein were then analyzed in the esophageal carcinoma cells. The results showed that 5-FAM-labeled recombinant NGAL protein could internalize into the EC1.71 and EC109 cells. Furthermore, the internalized NGAL protein could induce the alteration of cellular morphology, resulting in generation of autophagosome, transcriptional up-regulation of genes associated with autophagy and increase of phospho-ERK1/2 (p-ERK1/2). Interestingly, the treatment with the NGAL protein did not affect the intracellular iron level. These data indicate that induced autophagy by exogenous NGAL protein is a mechanism that internalized NGAL plays important roles in esophageal carcinoma cells, independent with NGAL-mediated iron transport process, while ERK1/2 signal pathway is involved in activation of autophagy by exogenous NGAL protein.]]> 2009/3/10 0:00:00 ZHANG Pi-Xian,FANG Wang-Kai,XU Li-Yan,JIANG Ji-Kai,SHEN Zhong-Ying,DU Ze-Peng,LU Xiao-Feng,ZHOU Fei,XIE Jian-Jun,WU Bing-Li,CUI You-Hong,XIE Dong and LI En-Min 892true 2009/3/26 0:00:00 DONG Guo-Zhen,YANG Zhi,ZHANG Zhao-Qi,YU Wei,LI Jing-Wei,HE Yi,ZHANG John X and WENG Xu-Chu 891true NCT), its 5′-flanking region has been characterized and a 270 bp fragment containing the TSS(transcription start site) for the promoter activity has been identified. EMSA assays confirmed that all four AP-1 binding sites and two NFAT sites in the NCT promoter region were able to bind relative transcription factors in vitro. Mutations, as well as treatment with PDTC, which adjust the regulatory effect of AP-1 and NFAT, altered NCT promoter activity in both HeLa cells and rat cortical neurons. The results demonstrated that AP-1 and NFAT are involved in the regulation of hNCT transcription and suggest that balanced activation of AP-1 and NFAT ensures a strict temporal and tissue-specific control of NCT transcription.]]> 2009/3/26 0:00:00 YANG Mei,CAI Fang,PAN Qian,LONG Zhi-Gao,XIA Jia-Hui,XIA Kun and ZHANG Zhuo-Hua 890true in vivo. TNF-α reporter gene activity results found that protein complex of p38-STAT3 and ERK2-STAT3 coordinately increased TNF-α activity. After STAT3 was interfered, the TNF-α activity markedly decreased. These data indicated that there exists interaction between p38 and STAT3 protein, ERK2 and STAT3 protein. The complex of the proteins can coordinately regulate TNF-α expression. After interfereing STAT3 pathway, the coordinated action on TNF-α transcription activity might be obviously reduced.]]> 2009/5/5 0:00:00 YANG Li-Ping,Yao Yong-Ming,LI Jie-Ping,YE Qi-Nong and SHENG Zhi-Yong 889true 2009/5/27 0:00:00 LI Yin-Tao,WU Wei-Bin,HONG Yi,WANG Wen-Zhong,YANG Jun-Wu,XIE Jian-Hui and WU Xing-Zhong 888true 2009/3/11 0:00:00 ZHANG Xin,MIAO Xiang-Yang,YIN Xun－He,MA Yan－Fang,QU Zhao-Jie and ZHANG Qiu-Ting 887true 2009/4/8 0:00:00 WU Yi-Bo,BO Xiao-Chen,YAN Li-Rong,YU Guang-Chuang,LIU Hui,SUN Han-Chang,XIE Hong-Wei and WANG Seng-Qi 886true 2009/3/11 0:00:00 CHEN Wei,LUO Liao-Fu,ZHANG Li-Rong and XING Yong-Qiang 885true 2009/2/24 0:00:00 SUN Hai-Yan,SUN Xin-De and ZHANG Ji-Ping 884true 2009/3/12 0:00:00 YU Chao,LI Gang,ZHOU Li-Na,WANG Shu,WANG Yong-Ming and LIN Ling 883true in vitro, and explore its probable mechanism, the nanoprodrug PASP-MTI was synthesized by DL-aspartic acid and metronidazole in a relatively mild condition. Its structural characteristics were attested by means of infrared spectrum (IR), 1H nuclear magnetic resonance (1H-NMR), transmission electron microscope(TEM) etc measurements. The release behavior of metronidazole in PASP-MTI nanoprodrug was investigated by the dialysis method. The MTT assay was employed to test PASP-MTI’s trichomonacidal effect in vitro and the apoptotic-like processes of trichomonas were detected by fluorescence microscope and flow cytometer (FCM). The IR and 1H-NMR showed that the metronidazole was conjugated to poly aspartic acid high molecular chains by ester bonds. Synthesized PASP-MTI nanoprodrug was globular and had a mean diameter 404.8 nm with favorable dispersity. The drug loading was about 30% and the cumulative release profile was delayed (about 38.01% of MTI loading in the PASP-MTI nanoprodrug was released within the first 24 h and 86.64% in 30 d, while the release of free metronidazole was almost complete by 24 h). The trichomonacidal activity of PASP-MTI nanoprodrug increased remarkably{IC50 of PASP-MTI and free metronidazole was 1.22 mg/L and 2.85 mg/L, respectively}. Observed by fluorescence microscope, the nuclei of trichomonas presented a series of changes similar to apoptosis such as chromatin agglutination, nuclear fragmentation, etc. The FCM study indicated that the apoptotic rate in the PASP-MTI group was the highest of all groups, up to 35.69%. It is concluded that PASP can be effectively used in drug delivery systems, the synthetic novel poly aspartic acid-metronidazole nanoprodrug has higher drug loading, better the role of prolonged-release and higher trichomonacidal effect in vitro. Its mechanism may be related with enhancing the polymer prodrug’s internalization and inducing the apoptosis of trichomonas.]]> 2009/3/27 0:00:00 QI Xiao-Hong,CHEN Xi-Min,FENG Zhen-Qing,GUAN Xiao-Hong,WU Jun,CHEN Qiang,KAN Yan-Jing and TONG Hua 882true amide) and temporin-Lb(LFRHVVKIFEKYLamide) were cloned from Lithobates catesbeianus. Synthetic peptides of temporin-La and temporin-Lb showed strong antimicrobial activities against bacteria tested, especially Gram-positive bacteria. Besides, temporin-La showed no haemolytic activity to rabbit erythrocytes at the concentration of 250 mg/L while temporin-Lb showed weak haemolytic activity(LC50 ≈ 230 μmol/L). Transmission electron microscopy showed that temporin-La and temporin-Lb induced different effects on bacterial structure of Staphylococcus aureus.]]> 2009/5/14 0:00:00 ZHAO Rui-Li,HAN Jun-You,HAN Wen-Yu,LEI Lian-Cheng,SUN Chang-Jiang,FENG Xin,JIANG Li-Na,QIAO Hong-Wei and CAI Lin-Jun 881true P < 0.001),especially in 72 hours; down-regulation of GnT-Ⅴ expression can enhance adhesive ability(t=-3.357, P < 0.01) and inhibit chemotactic migration(t=44.051, P < 0.001) in LoVo cell line; quantitative analysis of the wound closure assay also indicated that down-regulation of GnT-Ⅴ expression can significantly prolong wound heal hours of LoVo cell line; cell invasive experiment using Matrigel gel showed that penetrative cell numbers of LoVo GnT-Ⅴ/1564 and LoVo GnT-Ⅴ/NC were 5.10±1.25 and 39.55±2.16 respectively, penetrative cell numbers of LoVo GnT-Ⅴ/1564 cell was reduced obviously, compared to negative control group cell (t=61.626, P < 0.001). The shRNA aimed at GnT-Ⅴ gene could reduce the expression of GnT-Ⅴ both in the level of mRNA and protein. By this way, it can inhibit proliferation, migration and invasion of LoVo cell line, so the sequence of RNA interference against GnT-Ⅴ may be a valid target to treat colorectal cancer.]]> 2009/3/30 0:00:00 HE Fu-Li,MA Qiang and ZHANG Jian 880true 2009/2/3 0:00:00 SHANG Yu,XU Xia-Lian,XIN Hong and CHANG Zhi-Jie 879true + cells, while CD45- cells have no capacity to form hematopoietic colony at all. The FACS sorted CD49f+/Sca-1+ cells have no difference of hepatic differentiation potential compared with whole fetal liver cells. The clonality was confirmed by cell mixing assay. Taken together, the HPP-CFC may represent a novel clonal model for hepatic differentiation from the blood cells in the mouse feta liver and will shed light on the associations underlying the hepatic and hematopoietic development.]]> 2009/2/2 0:00:00 ZHOU Jun-Nian,WANG Yun-Fang,YAO Hui-Yu,HE Wen-Yan,CHEN Hai-Xu,LI Si-Ting,SHI Yan,SHI Shuang-Shuang,NAN Xue,BAI Ci-Xian,LIU Bing,YUE Wen,MAO Ning and PEI Xue-Tao 878true 2009/4/24 0:00:00 ZHAO Wen-Xiu,WU Hong and TAN De-Yong 877true 2009/2/23 0:00:00 LI Xiu-Nan,LI Li,Yang Chuan-Jie,HAO Chun,QIAO Cha,YANG Fen,ZHOU Ping-Kun,WANG Pei,YUAN Zeng-Qiang,LI Yan-Ling,LU Wei-Yuan,ZHANG Wei-Yuan and DING Ku-Ke 876true E. coli cells, the effects of the recombinant KGF-2 protein in cultured keratinocyte were analyzed by using MTT assay and in situ TUNEL assay. Interestingly, results revealed that KGF-2 promoted keratinocyte cell growth by stimulating cell proliferation and attenuating cell apoptosis. These findings supported a few evidences that KGF-2 could contribute to alveolar epithelial cells against apoptosis. Cell migration assays for the first time revealed that KGF-2 could stimulate keratinocyte cell migration in vitro. In addition, in the pilot animal test, recombinant KGF-2 incorporated within the hydrogel dressing exhibited significantly stimulatory effect on cutaneous wound healing. These combined effects implicate a potential therapeutic application of human recombinant KGF-2 in the future.]]> 2009/4/13 0:00:00 CHEN Xia,HU Wei,WEI Wan-Gui,SHEN Yan,CHEN Yan,YANG Sheng-Li and GONG Yi 875true 2009/2/23 0:00:00 ZUO Yong-Chun and LI Qian-Zhong 874true HGPRT is decreased or even absent in these diseases. Rabbits are an ideal model for studying the pathology of these diseases. Therefore, the development of an HGPRT-knockdown rabbit model will be highly beneficial in such studies. Stable HGPRT-knockdown transgenic fibroblast lines were generated by transfecting rabbit fibroblasts with RNA interference (RNAi) plasmids. Polymerase chain reaction (PCR) analyses indicated that the average positive rate was 83.3%. The mRNA and protein levels of HGPRT in the transgenic fibroblast lines were significantly lower than that in the control. Transgenic rabbit blastocysts were derived after performing nuclear transfer. The results show that RNAi can be used to stably knock down expression of the HGPRT in rabbit fibroblasts and further improvements in related technologies will facilitate the use of this method for the generation of HGPRT-knockdown rabbits.]]> 2009/1/10 0:00:00 GUO Yi,ZHANG Chuan-Shan,LI Shan-Gang,LI Feng,GU Rui-Huan,XING Feng-Ying,LI Yao,YAO Gang and CHEN Xue-Jin 873true in vitro cell invasion, and the activity of NF-κB signaling pathway were analyzed in the transfected cell lines. The results showed that RKIP was significantly downregulated in 5-8F compared with 6-10B, in NPC compared with NNET, and not detectable in NPC metastasis. Overexpressed RKIP in 5-8F could decrease its in vitro cell invasion, whereas downregulated RKIP in 6-10B could increase its in vitro cell invasion. Overexpressed RKIP in 5-8F could decrease phosphorylated-IκB-α level and tansactivation activity of NF-κB, whereas downregulated RKIP in 6-10B could increase phosphorylated-IκB-α level and tansactivation activity of NF-κB. Taken together, the results suggest that RKIP may be a NPC cell metastasis suppressor, and decreased RKIP expression is associated with the increased metastasis capability of NPC cells possibly through the activation of NF-κB signaling pathway.]]> 2008/12/22 0:00:00 CHEN Yan,LI Jun,YI Hong,OUYANG Guo-Liang,LI Cui,ZHANG Peng-Fei,LI Mao-Yu,PENG Fang,CHEN Zhu-Chu,LI Jian-Ling and XIAO Zhi-Qiang 872true 2+ regulates the activity of many enzymes and proteins, but also free-CaM (no Ca2+ bound) and Ca2+-independent CaM-binding proteins play roles in plant and animal cells. There is no in vivo method to identify the interaction between free-CaM and Ca2+-independent CaM-binding protein (CaMBP). Using site-directed mutagenesis by polymerase chain reaction (PCR), 5 mutant Arabidopsis calmodulin isoform 2 (AtCaM2) genes, mCaM21, mCaM212, mCaM2123, mCaM2124 and mCaM21234 were obtained. The mutant mCaM2 encoded glutamine in place of glutamate (E32Q; E68Q; E105Q; E141Q) in one or more EF-hand Ca2+-binding motifs of AtCaM2. The recombinant mCaM2 proteins were produced in Escherichia coli, and subsequently separated on SDS-PAGE in the presence of Ca2+ or EGTA, their electrophoresis mobilities were related with that of mutant EF-hand motifs. 45Ca2+ overlay analysis indicated that the more glutamate replaced by glutamine, the lower affinity with Ca2+ in the mCaM2 proteins. The mCaM21234 mutant protein (E32Q; E68Q; E105Q;E141Q) was unable to bind Ca2+. Using yeast two-hybrid technique with mCaM21234 as bait, it was possible to see interaction in Arabidopsis of AtCaM2 with IQD26, a calcium-independent CaM-binding protein. Site-directed mutation of AtCaM2 will aid the research of Ca2+, CaM and Ca2+-independent CaMBPs in plant biological processes.]]> 2009/2/10 0:00:00 GAO Li,WANG Zhen-Jie and CUI Su-Juan 871true P < 0.05), 100 nmol/L pGhrelin significantly promoted porcine preadipocyte proliferation compared to control group (P < 0.01). pGhrelin downregulated Caspase-3 activity and expression mRNA of porcine preadipocyte, induced preadipocyte proliferation and inhibited apoptosis. Caspase-3 dependent apoptosis regulation signalling was supposed to be involved in the apoptosis mechanism.]]> 2009/3/11 0:00:00 TANG Sheng-Qiu,SHU Gang,ZHU Xiao-Tong,WANG Song-Bo,GAO Ping,WANG Xiu-Qi,ZHANG Yong-Liang and JIANG Qing-Yan 870true LRRC4, leucine-rich repeat C4 protein, is a new member of leucine-rich repeat (LRR) superfamily. It is a novel tumor suppressor. LRRC4 inactivation is commonly found in glioma cell lines and primary glioma biopsies. Previous study did not find any genetic alteration of LRRC4 in primary glioma. In order to explore an alternative mechanism underlying inactivation of LRRC4 in glioma, glioma cell lines SF126 and SF767 were treated by methylase inhibitor 5-Aza-2′-deoxycytidine (5-Aza-CdR). Methylation-specific PCR was used to examine the methylation status changes of LRRC4 promoter in SF126 and SF767 cell lines. LRRC4 mRNA expression in SF126 and SF767 cell lines treated by 5-Aza-CdR was detected by RT-PCR. In addition, the effect on glioma cell lines cell growth and cell cycle of 5-Aza-CdR were assayed by MTT and FCM. The results indicated that LRRC4 promter was completely methylated in SF126 and SF767 cells. However, LRRC4 promoter aberrant hypermethylation can be reversed and LRRC4 expression can be induced by 5-Aza-CdR. Moreover, 5-Aza-CdR displayed a growth inhitory effect on SF126 and SF767 cells in a dose- and time-dependent manner after exposure to 5-Aza-CdR at different concentration for different time. FCM analysis showed that SF126 and SF767 cells cell cycles were blocked at G0/G1 phase after 5-Aza-CdR treatment for three days. Taken together, glioma cell lines SF126 and SF767 cell growth could be inhibited and cell cycles could be blocked by 5-Aza-CdR; promoter hypermethylation is the important mechanism of LRRC4 inacctivation in glioma cell lines; methylation can be reversed and LRRC4 expression can be induced by 5-Aza-CdR. All this suggest that LRRC4 may serve as a demethylation therapeutic target in glioma.]]> 2009/3/27 0:00:00 ZHANG Zu-Ping,WU Ming-Hua,TANG Hai-lin,WANG Rong,LI Dan,LI Xiao-ling and LI Gui-yuan 869true P < 0.05). So it was concluded that cytoplasmic M-CSF accelerates the proliferation of HeLa cells by up-regulating the mRNA expression of cyclinD1/D3 and CDK2/6.]]> 2009/3/30 0:00:00 TU Jian,WU Hai-Yan,ZHANG Meng-Xia,ZHANG Xiao-Hong,LUO Hong-Mei,LONG Zhi-Feng,WANG Yu-Hua,LEI Xiao-Yong and TANG Sheng-Song 868true 2009/3/26 0:00:00 LI Miao,WANG Yu-Fei,WANG Yu,GAO Hui,LI Na,SUN Yuan,LIANG Bing-Bing and QIU Hua-Ji 867true Escherichia coli biotin carboxyl carrier protein (BCCP87) was expressed in E. coli using fusion expression strategy. Results from Western blot analysis and SDS-PAGE analysis showed that the fusion protein could be biotinylated in vivo, and the molecular mass of the fusion protein was about 64 ku. The biotinylated ATP sulfurylase could be immobilized on the surface of magnetic beads coated with strepavidin, and the immobilized ATPS could be used for quantification of PPi and pyrosequencing. An effective enzyme for the large-scale chip-based pyrosequencing system was supplied.]]> 2009/3/11 0:00:00 ZOU Bing-Jie,LUO Juan,WU Hai-Ping and ZHOU Guo-Hua 866true 2009/3/11 0:00:00 ZHANG Jian,HAN Ya-Ling,KANG Jian,ZHANG Xiao-Lin,QI Yan-Mei,YAN Cheng-Hui and LI Shao-Hua 865true Gloydius blomhoffi brevicaudus in the laboratory. Previous researches had proved that rAdinbitor could inhibit proliferation of C6 glioma cells as well as promote their apoptosis. The molecular mechanism of rAdinbitor’s effects on C6 cells need to be further studied. rAdinbitor was expressed in E. coli BL21/pET23b-adinbitor and purified with Ni Sepharose 6 Fast Flow. The purified protein was confirmed by Western blotting. C6 cells were induced with fibronectin (FN). The effects of rAdinbitor with different concentrations on the expression of FAK, MEK1/2 and Caspase-3 as well as on activity of FAK and ERK1/2 in FN-induced C6 cells were studied by immunoblotting and immunoprecipitation. Results showed that rAdinbitor with different concentrations could obviously reduce the expression of FAK and MEK1/2, increase the expression of Caspase-3, as well as decrease ERK1/2 phosphorylation; besides 10 mg/L rAdinbitor, other concentrations’ rAdinbitor could inhibit FAK phosphorylation obviously. All those effects were dose-dependent. Results indicate that the effects of rAdinbitor on decreasing expression and activity of FAK and inhibiting Ras-MAPK signaling pathway play an important role in suppressing the proliferation of C6. Furthermore, the increase in Caspase-3 expression implies that the increase in apoptosis of C6 cells might be due to the suppression of rAdinbitor on the activity of ILK and PI-3K/Akt pathway.]]> 2009/1/8 0:00:00 ZHAO Ting,LI Jin-Ping,HU Yan-Rong,HONG Yan and ZHAO Bao-Chang 864true 2009/4/13 0:00:00 XIANG Bo,WANG Li,YI Mei,OUYANG Jue,LI Xia-Yu,ZHANG Zu-Ping,LI Xiao-Ling and LI Gui-Yuan 863true Deinococcus radiodurans, the carotenoids of this bacterium were extracted with cold methanol and acetone, then were isolated and analyzed by LC-MS technique. Two colorless mutants knocking out of the phytoene synthase (crtB) and phytoene desaturase (crtI) genes were constructed by PCR and in vivo homologous recombination. The successful construction was confirmed by phenotype observation and carotenoids analysis by high performance liquid chromatography (HPLC). Comparative analysis of the sensitivities among the two colorless mutants and wild R1 showed that the two colorless mutants were more sensitive to ionizing radiation (IR) and ultraviolet (UV), indicating that the deletion of crtB and crtI genes made the mutants unable to biosynthesize the crucial intermediate lycopene and series of downstreaming carotenoids. The scavenging abilities of the two carotenoids were measured using the ultraviolet induction of bacteriophage λ (UIB) system and electron spin resonance (ESR) spin trapping technique in vivo and in vitro, respectively. The results showed that the main two carotenoids exhibited significant antioxidant capacities to superoxide anion (O2·) and hydroxyl radicals (·OH) which were two main types of free radicals produced during IR. This study has provided for the first time a new and direct evidence to explore the biosynthesis genes of carotenoids and their biological functions for the radioresistant mechanisms of D. radiodurans.]]> 2008/12/18 0:00:00 YANG Qiao,ZHANG Xiao-Ling,ZHANG Lei,DAI Jun,ZHANG Jun-Xiang and JIAO Bing-Hua 862true Helicobacter pylori(H. pylori) on cell gap junction ultrastructure of gastric epithelial cells, and to explore carcinogenic mechanism of H. pylori from the changes of cell gap junction, BGC-823 cells were co-cultured with different H. pylori strains for 24 h and 48 h. The cell gap junction ultrastructure was observed under transmission electron microscope with sample preparation of fixation and embedding in situ. In 70 patients with gastric cancer(GC), H. pylori was detected by rapid urease test, basic fuchsin stain and 14C-urea breath test. The CagA gene of H. pylori was determined by PCR and the cell gap junction ultrastructure was observed under transmission electron microscope. More cell gap junctions and junction complexes of BGC-823 cells were found in control group without H. pylori. Groups with H. pylori had less number of cell gap junctions, less number of junctions/ unit perimeter, shorter length of junctions /unit perimeter, and bigger width of the intercellular space, comparing to control groups without H. pylori(P < 0.001 or P < 0.005). The number of cell junctions and the number of junctions/unit perimeter in the groups co-cultured with NCTC J99, GC 01 and NCTC 11639(CagA+) were less than that in the groups co-cultured with NCTC 12908(CagA-) (P < 0.001 or P < 0.05), and the length of junctions/unit perimeter in the groups co-cultured with NCTC J99 and GC 01 was shorter than that in the groups co-cultured with NCTC 12908 (P < 0.001). In patients with GC, the number of cell junctions, the number of junctions/unit perimeter and the length of junctions/unit perimeter in group H. pylori infection were all less than those in group without H. pylori infection(P < 0.001), and that in CagA+ H. pylori group were less than that in CagA- H. pylori group, but its smallest width of the intercellular space was longer than that in CagA- H. pylori group. The above results showed that the changes of cell gap junction of gastric epithelial cells were associated with H. pylori infection especially CagA+ H. pylori infection.]]> 2009/1/10 0:00:00 XU Can-Xia,JIA Yan,YANG Wen-Bin,ZOU Hui-Fang,WANG Fen and SHEN Shou- Rong 861true 2008/12/29 0:00:00 SUN Shu-Feng,ZHANG Kai,XU Wei,WANG Gang,CHEN Jian-Jun and SUN Fei 860true in vitro cytotoxicity assay. The isotype and specificity of resultant clone was determined, and its efficacy to neutralize the activity of purified Stx2 was evaluated by in vitro and in vivo toxicity model. Lastly, its spectrum of activity against Stx2 variants was also accessed by mouse toxicity model. It was demonstrated that one of the 12 positive MAb clones against Stx2, designating S2C4 had neutralizing activity. S2C4 belongs to the immunoglobulin G1 subclass and has a κ light chain, and it reacts with the A subunit of Stx2 and does not bind to Stx2 B subunit or to Stx1. S2C4 could efficiently neutralize the cytotoxicity of Stx2 to Vero cells and mice. It also protected mice against lethal doses of Stx2 variants challenge including Stx2c and Stx2vha. S2C4 is a promising candidate molecule in preventing the progression of hemolytic-uremic syndrome (HUS) mediated mainly by Stx2 in Stx-producing Escherichia coli (STEC) infection.]]> 2008/12/19 0:00:00 JIAO Yong-Jun,ZENG Xiao-Yan,GUO Xi-Ling,SHI Zhi-Yang,FENG Zhen-Qing and WANG Hua 859true vs 28%, χ2=28; P < 0.05). The results of RT-PCR and immunohistochemical staining discovered that 14-3-3σ gene exhaustive methylation leads to absent 14-3-3σ mRNA as well as protein expression in nasopharyngeal carcinoma tissues and normal nasopharyngeal mucosal tissues. Tissues characterized with partial methylation 14-3-3σ gene represented remarkable down-regulated 14-3-3σ mRNA and protein expression level when compared with tissues characterized with unmethylation 14-3-3σ gene. Graded correlation analysis discovered that 14-3-3σ gene hypermethylation status is positively correlated with NPC lymphoid node metastasis and NPC clinical staging. The research demonstrated that 14-3-3σ gene was frequently hypermethylated in nasopharyngeal carcinoma tissues and leads to decreased or depleted gene expression, 14-3-3σ expression level was correlated to nasopharyngeal carcinoma lymphoid node metastasis and NPC clinical staging.]]> 2009/1/6 0:00:00 TAN Shuang-Xiang,LI Jun,YI Hong,TANG Cen-E,CHENG Ai-Lan,CHEN Zhu-Chu,LI Jian-Ling and XIAO Zhi-Qiang 858true 2009/3/11 0:00:00 ZHU Ning,MA Jun,ZENG Shao-Ju,LIN Yu-Tao,ZHANG Xin-Wen and ZUO Ming-Xue 857true P < 0.05). PLGA encapsulating plasmid microspheres were prepared and injected intramuscularly into rabbit legs. Growth behavior and IGF-1 level were measured at day 0, 15, 30 and 45 after injection. Greater body weights gain and higher serum IGF-Ⅰlevels were observed in three plasmid microsphere injection groups, compared with control group. At day 30, the body weight gain in P-G-P group was greater than saline group (81%, P < 0.01), P-G microsphere group (15%, P < 0.05) and P-P microsphere group (7%, P > 0.05), serum IGF-Ⅰ concentration in P-G-P microsphere group showed a 16.68% increase to P-G microsphere (P > 0.05), a 17.14% increase to P-P microsphere(P > 0.05) and a 50.46% increase to control (P < 0.05). These results suggest that co-expression of GHRH and PACAP in one expression plasmid might exert an additive stimulation of GH secretion and growth when delivered into rabbit skeletal muscle with PLGA microsphere. The results may provide a new approach to regulate animal growth.]]> 2009/1/10 0:00:00 LIU Song-Cai,DAI Jian-Wei,REN Xiao-Hui,HAO Lin-Lin and Zhang Yong-Liang 856true 2009/2/10 0:00:00 ZHANG Heng,WANG Ge-Fei,GAO Yuan-Li,CHEN Xiao-Xuan,LIN Gui-Mei and LI Kang-Sheng 855true in vitro culture. The effects of IGF-1 in different concentrations was determined on the development of blastocyst in hyperglycemic conditions (30.0 mol/L glucose) in vitro, as well as the apoptosis of embyro cells by using nuclear DNA double-dyed assays. Moreover, the expression of igf-1 and other genes associated with apoptosis in vitro embryo was detcted by Real-time quantitative PCR. The results showed that the total cell number of blastocyst fell off along with the increasing of glucose concentration. High concentrations of glucose (≥30 mmol/L) could significantly inhibit the growth of embryo cells (P < 0.01). The results of real-time quantitative PCR showed that the expression of igf-1 was down-regulated in mouse blastocyst with pregnancy associated with diabetes, and positively correlated with the concentration of glucose. The expressions of apoptosis-related genes bcl-2 and bcl-xl were up-regulated along with the increasing of IGF-1 concentration, while the p53 gene and the apoptosis-related gene Bax were down-regulated. The embryo cells apoptosis had been gradually reduced with the increasing of IGF-1 concentration in vitro, and in the IGF-1 concentration of 100 μg/L, there was almost no apoptosis in embryo cells. These experiments demonstrated that hyperglycemic conditions in vitro can inhibit the growth and development of the embryo cells resulting in the down-regulated expression of igf-1, and IGF-1 could inhibit the apoptosis of blastocyst and be benefit for the growth of blastocyst.]]> 2009/2/24 0:00:00 LUO Xiao-Min,ZOU Hai-Yan,ZHANG Zhe,PAN Ai-Hua,YIN Fang,WEI Miao,YANG Jing and HUANG He 854true Chlamydia trachomatis(C. trachomatis) serovar D genome was amplified and cloned into the pGEX and pDSRedC1 vectors. The recombinant plasmid pGEX-CT358 was constructed and expressed as GST fusion proteins. The GST-CT358 fusion protein was used to immunize mice to raise the antibodies, which specifically recognized CT358 without cross-reacting with other unrelated proteins. The antibodies were then used to localize the endogenous CT358 protein and determine the expression pattern in Chlamydial infected cells using an indirect immunofluorescence assay (IFA). Meanwhile, pDSRedC1-CT358 recombinant plasmid was transfected to HeLa cells to evaluate the effect of CT358 expression on the subsequent chlamydial infection. The hypothetical protein CT358 was identified in the inclusion membrane of C. trachomatis-infected cells for the first time,and it was detected as early as 12 h after C. trachomatis infection and remained in the inclusion membrane throughout the rest of the infection cycle. Cytosolic expression of CT358 via a transgene failed to affect the subsequent chlamydial infection. These observations together have demonstrated that CT358 is a newly identified chlamydial inclusion membrane protein, giving the potentially importance for further understanding the mechanisms of chlamydial intracellular parasitism.]]> 2009/1/10 0:00:00 LI Zhong-Yu,WU Yi-Mou,HUANG Qiu-Lin,WANG Shi-Ping and ZHONG Guang-Ming 853true Agrobacterium tumefaciens possesses many advantages as a model bacterium for the study of a wide variety of biological processes. Gene disruption or inactivation is a powerful and direct tool for investigation of in vivo gene functions. The intensive study of A. tumefaciens has increased the need for simple and highly efficient procedures to manipulate its genome. The sacB gene was used as a counterselectable marker to develop a gene replacement procedure that allows precise insertion, deletion, and allele substitution of any gene sequence in A. tumefaciens without altering the genome in any other way. A kanamycin resistance (KmR) cassette was constructed to the suicide vector as the positive selection marker. The suicide plasmid containing DNA fragments homologous to the flanking sequences of the target gene was integrated into the recipient cell genome at the target gene locus by intermolecular homologous recombination, generating the KmR-single cross-over colonies. The effect of homologous sequence length on the intermolecular homologous recombination was analyzed. The second cross-over colonies generated by intramolecular homologous recombination occurring between two tandem repeats were simply screened out by counter-selection of sacB. Data showed that the intervening sequence length between two repeats significantly affected the intramolecular homologous recombination frequency in A. tumefaciens, indicating that A. tumefaciens adopted the homologous recombination mechanism similar to that in E. coli. All these results demonstrated that investigators could minimize the numbers of colonies to be analyzed and reduce the overall workload by optimizing the relative length of two homologous fragments and using the specific type of single cross-over transformants for screening the second cross-over event. This mutagenesis strategy had successfully been used to generate the double unmarked Δvbp2Δvbp3 mutant in two A. tumefaciens strains.]]> 2008/10/24 0:00:00 GUO Min-Liang,ZHU Qing and GAO Dian-Kun 852true P < 0.01), and individuals with genotype AA of the SNP4 had the highest cashmere weight compared with those with the other genotypes (P < 0.05). The relationship between diplotype and cashmere fiber traits was also analyzed, the results showed that both cashmere fiber diameter and cashmere weight were significantly correlated with the diplotype H7H7 (P < 0.05). These results suggested that IGF-Ⅰ gene would be useful candidate gene in selection program on cashmere fiber traits in cashmere goats.]]> 2008/10/29 0:00:00 GUO Cui-Hua,JIA Cun-Ling,ZHANG Wei,ZHU Xiao-Ping and JIA Zhi-Hai 851true in vitro could simulate the pathological characteristics of osteomyelitis patients. The results of quantitative real-time PCR and Western blot showed that CHI3L1 was up-regulated obviously in the infected bone tissues of osteomyelitis patients and LPS-stimulated osteoblasts. Analysis of the luciferase activity of NF-κB reporter gene vector revealed that LPS could activate NF-κB. Bay11-7082, an inhibitor of NF-κB activation, suppressed the elevation of CHI3L1 expression induced by LPS. Pre-incubation of osteoblasts with anti-TNF-α antibody or silencing TNF-α receptor expression by siRNA inhibited the induction effect of LPS on CHI3L1. Inhibition of NF-κB activation also prevented up-regulation of TNF-α induced by LPS. In conclusion, LPS stimulated TNF-α expression through activating NF-κB, then TNF-α induced CHI3L1 expression. It was demonstrated for the first time that CHI3L1 expression is promoted in osteomyelitis and LPS-treated osteoblasts and investigates the molecular mechanism of LPS-induced CHI3L1 expression in osteoblasts.]]> 2008/10/21 0:00:00 GAO Lei and CAI Guo-Ping 850true Δ252～351 retroviruses. Therefore, the NP69-LMP1 and NP69-LMP1Δ252～351 cell lines of stable expression LMP1 were established. Sequentially, cellular proliferation of the NP69-LMP1 and NP69-LMP1Δ252～351 cell lines was compared to draw growth curve, experiment plate clone formation and test soft agar colony forming. Meanwhile, adopted proteomic methods, the differential expression proteins were identified between NP69-LMP1and NP69-LMP1Δ252～351 cell lines, and the expression levels of partial identified proteins were verified by Western blotting and fluorescent real-time quantitative RT-PCR. The results showed: 1) the ability of LMP1Δ252～351 promoting NP69 cell proliferation was obviously decreased to compare with wild type LMP1(n=3，P < 0.05). 2) 16 proteins, 8 up- and 8 down-regulated ones, of LMP1-CTAR3 mediated regulation were identified from infected NP69 cell lines. 3) The differential expression of partial identified proteins was similar with 2-DE separated ones and confirmed by Western blotting and fluorescent real-time quantitative RT-PCR. These results suggested that LMP1-CTAR3 is the important activating domain of inducing cellular proliferation and the domain probably plays the role of mediating to regulate the expression of G-protein, isocitric enzyme, and so on.]]> 2008/10/21 0:00:00 Zhang Zhi-Wei,ZHANG Qiong,YU Yan-Hui,OUYANG Yong-Mei and He Zhi-Min 849true via a cellular process to induce gene silence through degradation of homologous transcripts. The experiments were designed to inhibit proliferation of human retinoblastoma cells by knockdown human transcription factor E2F1 expression level using RNA interference technology. Three different hairpin shRNA inserts were designed targeting human transcription factor E2F1. The three inserts were synthesized and subcloned into a shRNA espression vector pSilencer H1puro which contains the H1 RNA polymerase Ⅲ promoter. The three plasmids encoding E2F1 short hairpin were transfected into human retinoblastoma cells line HXO-Rb-44 and human liver cancer cell line SMMC-7721 in vitro. Real-time RT-PCR was used to measure E2F1 mRNA level. For function analysis, propidium iodide (PI)-stained DNA content method was used to identify cell cycle by flow cytometer. The selected shE2F1 were cloned into an adenovirus system and packaged to adenovirus. Retinoblastoma HXO-Rb-44-gfp cell line which has been marked with green fluorescent protein (GFP) were infected by Ad-shE2F1 for 48h, and Ad-shNegative was used as control. The infected cells were injected into anterior chamber of nude mice eyes. The eyes were investigated and the consecutive and dynamic photos of the condition of RB tumor′s growth were taken by the special fluorescent microscope. One in three RNAi expression plasmids delivery resulted in 83.5% suppression of E2F1 mRNA transcription level. The same shRNA showed 89% inhibition in entering S phase. In nude mice eye anterior chamber, Ad-shE2F1 infected tumor cells grew more slowly than the negative control. The results showed that efficacy of Ad-shE2F1 were helpful in the process of limiting the growth of retinoblastoma. RNAi gene therapy directed against the human transcription factor E2F1 is a new therapeutic approach to suppress retinoblastoma cells proliferation.]]> 2008/11/18 0:00:00 WANG Feng,MA Jia-Lie,LI Hui-Ming,CHEN Xia-Fang,WANG Yi-Fei and HUANG Qian 848true 2008/12/5 0:00:00 ZHANG Xiao-Yi,HE Hong-Qiu,LIU Bin and Wang Cun-Xin 847true in vitro and enhance blood coagulation in vivo, which prove its biological activity. Using this recombinant mTF as immunogen, a murine monoclonal function-blocking antibody to mTF was also generated, which can inhibit mTF-initiated plasma clotting in vitro. And by treating with this function-blocking antibody, the thrombus formation in a murine deep vein thrombosis model was attenuated successfully, which suggests the important role of tissue factor in deep vein thrombosis. In all, with the active mTF recombinant protein and the mTF function-blocking antibody, the functional investigations of TF in murine models of various research areas become more convenient and feasible.]]> 2008/11/18 0:00:00 PEI Xin-Hui,CHI Shan,WANG Jin-Tao,LIN Zhi-Xin,ZHANG Xue-Hong and GENG Jian-Guo 846true 2008/12/17 0:00:00 FU Cong and LIN Kui 845true P < 0.05, P < 0.01, respectively). Expression of MT1-MMP, MT2-MMP and Ezrin proteins in the stageⅡ, stageⅢ and Ⅳ NPC and NPC with lymph node metastasis is significantly higher than that of in the stageⅠ NPC and NPC without lymph node metastasis (P < 0.05, P < 0.01 respectively), but expression of nm23-H1 protein in the stageⅡ, stageⅢ and Ⅳ NPC and NPC with lymph node metastasis is significantly lower than that of in the stageⅠ NPC and NPC without lymph node metastasis (P < 0.05). There were significantly positive correlation between MT1-MMP and MT2-MMP (r = 0.308, P < 0.001), nm23-H1 and E-cad(r = 0.167, P < 0.05), nm23-H1 and TIMP-2(r = 0.279, P = 0.001); E-cad and TIMP-2(r = 0.279, P = 0.001) in the NPC. Also, there were obviously negative correlation between MT1-MMP and E-cad(r = -0.188, P < 0.05), MT1-MM and TIMP-2(r = -0.233, P < 0.05), Ezrin and E-cad (r = -0.204, P < 0.05) in NPC. Based on the cluster analysis, there was common higher expression of MT1-MMP, MT2-MMP and Ezrin proteins in the NPC than that of in the chronic inflammatory nasopharyngeal epithelium (P < 0.05). However, common higher loss expression of nm23-H1, E-cad and TIMP-2 proteins were found in NPC compared with the peri-cancer nasopharyngeal epithelium and chronic inflammatory nasopharyngeal epithelium (P < 0.05, P < 0.01). Logistic regression analysis and validity estimation indicated that MT1-MMP gene might be better independent prognostic value for metastasis and clinical progress of NPC. The results suggested that high protein expression of multiple metastasis related-genes, low expression of the metastasis suppressor genes and loss balance between metastasis genes and metastasis suppressor genes might play important role in the metastasis and clinical progression of NPC. MT1-MMP protein can be used as the better independent prognostic molecular marker for metastasis of NPC.]]> 2009/4/3 0:00:00 FAN Song-Qing,ZHANG Wen-Ling,XU Li-Na,ZHOU Yan and LI Gui-Yuan 844true 2008/12/17 0:00:00 ZHU Yan-Xia,LIU Tian-Qing,SONG Ke-Dong,MA Xue-Hu and CUI Zhan-Feng 843true LCRG1(laryngeal carcinoma related gene1, LCRG1), a new candidate tumor suppressor gene of laryngeal carcinoma. However, it is known little about the possible regulatory mechanisms of LCRG1 gene expression. Restriction endonuclease digestion was used to obtain a set of the 5′, or 3′deletion mutants from the region(-169～+127) of the LCRG1 gene. It has been found that the minimal promoter of the LCRG1 gene is mapped at the region from -169～-57. Linker scanning mutational analysis in the region(-169～+127) of the LCRG1 gene was used to identify the crucial cis-elements within the promoter region, The key cis-elements are within the region from -137～-122. SP1, E2F1/DP1, EKLF and ZF9 transcription factor binding site sites were predicted in the region by bioinformatics analysis. Co-transfection with each of a panel of the expression plasmids of the known transcription factors with the relevant reporter construct indicates Sp1 is potent transcription factor for enhancement of the promoter activity, SP1 can also up-regulate the endogenous expression of LCRG1 gene. Electrophoretic mobility shift assay (EMSA) was applied to verify that the key cis-elements of LCRG1 gene exist sequence of Sp1 binding sites. The findings, which showed that the key cis-elements within the region from -137～-122 play an important role in expression of the LCRG1 gene, provide a novel evidence for further study of the function of LCRG1 gene.]]> 2009/2/10 0:00:00 XIE Hai-Long,CHEN Zhu-Chu,LI Jin-Hua,ZENG Long-Wu and TAN Gui-Huang 842true Escherichia coli is the most common gram-negative organism causing neonatal meningitis. In order to characterize the role of the pathogenicity island gene ibeT of Escherichia coli K1 in the pathogenesis of neonatal meningitis, an ibeT gene isogenic in-frame deletion mutant strain was constructed. Intracellular survival assay in human brain microvascular endothelial cells (HBMECs) showed that the deletion of ibeT inhibited the growth of Escherichia coli K1 within HBMECs. Confocal microscopy analysis showed that much more ibeT mutant remained in lysosomes of HBMECs compared with wild type Escherichia coli K1 after bacterial invasion into HBMECs. Transmission electron micrographs further showed that ibeT mutant strain was failed to disrupt the Escherichia coli containing vacuole (ECV) and escape into the cytoplasm. Furthermore, cytoplasmic buffering capacities of ibeT mutant were lower than wild type Escherichia coli K1 at pH 6.5 and pH 6.0. These results suggested that the expression of ibeT in Escherichia coli K1 contributed to ECV membrane degradation and subsequent escape from lysosomes into the cytoplasm for replication after Escherichia coli K1 invasion into HBMECs.]]> 2008/11/10 0:00:00 ZHANG Ke,ZHAO Wei-Dong,LI Qiang,FANG Wen-Gang,ZHU Li and CHEN Yu-Hua 841true Escherichia coli, purified, and used in a L929 cell killing assay. The assay results proved that the key amino acids for 9C6 binding were consistent with mouse TNF-α functional domain.]]> 2008/9/22 0:00:00 JIA Jun-Ying,ZHOU Hong-Zhe and TANG Jie 840true M 9.0～100.0 ku and pI 4～7. A total of 28 differentially expressed proteins were identified by matrix-assisted laser desorption ionization time of flight mass spectrometry(MALDI-TOF-MS), 5 of them were identified following NCBInr database queries, which are ubiqutin-protein ligase E2, putative glycine-rich protein, ascorbate peroxidase, putative cysteine proteinase inhibitor, ribulose bisphosphate carboxylase small chain clone 512. These proteins were involved in the process of energy metabolism, programmed cell death (PCD), regulating flower development.These results provide a clue for elucidating the mechanism of CMS.]]> 2008/10/6 0:00:00 CHEN Rui-Hong,YE Jing-Xiu,ZHANG Gai-Sheng,WANG Jun-Sheng,NIU Na,MA Shou-Cai,ZHAO Ji-Xin and ZHU Jian-Chu 839true +CHO- hybridoma cells. BALB/c mice were intraperitoneally injected with the selected hybridoma cells. Ascites were collected and monoclonal antibodies were purified using FPLC, and its Ig class, subclass, and titer were then determined respectively. The specificity of the yielded mAb was identified with CELISA, flow cytometry, ABC immunohistochemistry and immunoblotting. Detection of CELISA showed that 100 mg/kg dose of CP could tolerize the mouse to common epitopes shared between CHO and CHO-TM5 cells. And CELISA also discovered that all hybridomas positive for CHO-TM5 cells were negative for CHO cells. Five lines of positive hybridoma cells had been obtained altogether and 2F7 was selected randomly for next investigation. The Ig subclass of the mAb 2F7 was IgG1 and the titer of ascitic mAb was 1×10-6. Furthermore, the content of ascitic mAb was 19.56 g/L and chromosome numbers is 98. Flow cytometry, CELISA and Western blotting assays demonstrated that mAb 2F7 could specifically recognize hTM expressed on CHO-TM5 and human umbilical vascular endothelial cells (HUVEC). Meanwhile, the tissue specificity of mAb 2F7 was also identified by immunohistochemical ABC staining. On the other hand, Western blotting assays indicated that mAb 2F7 could recognize the antigen protein with 105 ku molecular mass under reduction condition. Moreover, the dissociation constant of mAb 2F7, 1.22×10-9 mol/L, indicated the affinity higher than some others. The results suggest that the immunotolerizing protocol provides a convenient general method for producing antibodies specific to desired protein isoforms. mAb 2F7 can specifically recognize the natural hTM expressed mainly on vascular endothelial cells, which will potentially useful for investigating the functions and clinic values of hTM.]]> 2009/2/9 0:00:00 GUO Zi-Fen,HE Shu-Ya,ZHU Bing-Yang,LI Bin-Yuan and LIAO Duan-Fang 838true 2008/11/3 0:00:00 LIU Ying-Fu,XIAO Zhi-Qiang,ZHANG Peng-Fei,LI Mao-Yu,LI Cui,PENG Fang,YU Yan-Hui,YI Hong and CHEN Zhu-Chu 837true 2008/11/11 0:00:00 XIE Si-Ming,DU Sha-Sha,FANG Wei-Yi and YAO Kai-Tai 836true 2008/10/29 0:00:00 BAI Yan-Yan,SHI Yi,HUI Guo-Zhen,ZHANG Yan-Rong,DONG Wan-Li,HU Jin and JIN You-Xin 835true 2+ and AMPK activity. To mimic this response, primary cultured rat skeletal muscle cells were treated with Caffeine to raise cytosolic Ca2+ and with AICAR to activate AMPK. The muscle cells were divided into different groups (Control, AICAR, Caffeine, AICAR/Caffeine, Caffeine+Compound C, AICAR/Caffeine+Compound C, AICAR+KN93, AICAR/Caffeine+KN93), which were used for experiments of stimulation by AICAR and Caffeine, inhibition by AMPK inhibitor, Compound C and CaMK inhibitor, KN93 respectively. The results showed that both AICAR and Caffeine induced about 2- and 3-fold increases respectively (P < 0.05, AICAR vs. Control; P < 0.05, Caffeine vs. Control) in GLUT4 mRNA in muscle cells, but the effect of GLUT4 mRNA induced by the combined stimulation of raising cytosolic Ca2+ and activating AMPK was not additive. Moreover, the Compound C, an AMPK inhibitor, decreased the Caffeine-induced increases in GLUT4 mRNA (P < 0.05, Caffeine+Compound C vs. Caffeine), and also attenuated an increase in GLUT4 mRNA induced by the combined stimulation of AICAR and Caffeine (P < 0.05, AICAR/Caffeine+Compound C vs. AICAR/Caffeine). Similarly, the Caffeine induced an increase in α1-AMPK phosphorylation (P < 0.05, Caffeine vs. Control) and furthermore the Compound C reduced apparently such an increase (P < 0.05, Caffeine+Compound C vs. Caffeine); however, KN93, a CaMK inhibitor, was completely able to inhibit the Caffeine-induced increase in GLUT4 mRNA, but failed to inhibit the AICAR-induced 2-fold increases in GLUT4 mRNA (P < 0.05, AICAR+KN93 vs. Control) and also failed to block an increase in GLUT4 mRNA induced by the combined stimulation of AICAR and Caffeine (P < 0.05, AICAR+Caffeine+KN93 vs. Control). These results demonstrate that the CaMK and AMPK signaling pathways are not complete independent, but are cooperative in the regulation of Caffeine-induced GLUT4 gene expression in cultured skeletal muscle cells. Collectively, based on our results and the other results this paper suggests that the GLUT4 gene expression in contracting skeletal muscle cells may be regulated through a CaMK-AMPK signaling pathway.]]> 2008/11/1 0:00:00 LI Liang-Gang and CHEN Huai-Qing 834true n=39) and the control group (NS, n=21). At 1.5 h, 3 h, 6 h and 12 h after the last seizure, apoptosis and autophagy were detected by transmission electronmicro-scopy (TEM) or in situ end labeling (TUNNEL). Electro-corticogram was observed after the last seizure. During P51 to P56, P81 to P84 and P92 to P95, the rats were tested for spatial learning and memory abilities with automatic Morris water maze task. On P95, mossy fiber sprouting and gene expressions in hippocampus were determined subsequently by Timm staining and real time RT-PCR methods. The results are as follows: a. TEM revealed the formation of autophagosomes and apoptosis in hippocampus after kindling. In the hippocampal neurons of the controls, chromatin and cytoplasmic organoids showed eumorphism with nuclear membrane integrity; 1.5 h after the last penicilin-induced recurrent seizures, “C”-morphous double membrane structure of dilated endocytoplasmic reticulum could be observed; 3 h after the last seizures, autophagosome appeared with manifest dilated endocytoplasmic reticulum; 6～12 h after the last seizures, the hippocampal neurons showed apoptotic feature such as nuclear chromatinic pyknosis and verge-aggregation. TUNNEL staining also showed elevated number of apoptosis neurons in hippocampus of RS group(P < 0.05). b. Vertex sharp transient wave and sharp wave were soon detected just in a few minutes afer the last seizure in RS group by Electrocorticogram. c. Escape latency. The escape latencies from the water maze were significantly longer in rats of RS group than that of the control at d5 of the first test, at d1 of the second test, and at d2 of the third test. d. Strategy analysis. RIDIT analysis showed that the scores were much worse in rats of RS group than that of the control at d4 and d5 of the first test. Moreover, the scores were much worse in rats of RS group than that of the control at the whole three days of both the second and third maze tests. e. Memory test. As for the value of distance in origin platform quadrant to total distance, the RS group seemed worse than the NS group in three maze tests(P < 0.05). f. Aberrant mossy fiber sprouting was seen in the inner molecular layer of the granule cells and the stratum pyramidale of CA3 subfield in RS group. g. Real time RT-PCR analysis showed decreased expression of CaMKⅡα and ZnT-1 in hippocampus of RS group than that in control group. Moreover, the four genes (CaMKⅡα, PRG-3, CCK and ZnT-3) in NS group and the two genes (CaMKⅡα and ZnT-1) in RS group showed copolymerization characteristic by cluster analysis. In addition, correlation and regression analysis demonstrated positive linear correlation among CaMKⅡα, CCK, PRG-3 and ZnT-3 in NS group and between CaMKⅡα and ZnT-1 in RS group. It was concluded that recurrent developmental kindling induced by penicillin could cause not only autophagy and apoptosis in the earlier stage of brain damage, but also have long- term effects on cognitive function and hippocampal mossy fiber sprouting which may be associated with the down-regulated expression of CaMKⅡα and ZnT-1 in hippocampus.]]> 2008/11/18 0:00:00 NI Hong,JIANG Yu-Wu,DING Mei-Li,WANG Zhe-Dong,TAO Lu-Yang and WU Xi-Ru 833true Cis9, trans11 and trans10, cis12-CLA are two major isomers of conjugated linoleic acid, which have stronger activities of anti-tumor. Based the previous studies, it was explored the pathway and mechanism of inducing apoptosis in human breast cancer cell line SKBR3 by c9, t11-CLA and t10, c12-CLA. It was confirmed that CLA could increase obviously the transcription and protein expression levels of PPARγ by RT-PCR and Western blot. The synchronism and correlation between PPARγ and apoptotic proteins Bax, Bcl-2, caspase3 changes were found with a dose- and time-dependent manner. PPARγ inhibitor GW9662 experimental result showed that there is cooperative relation between them. This is the first report that CLA induces apoptosis in SKBr3 cell by the signal pathway of PPARγ-Bcl-2-Caspase3. These observations indicated that CLA will be useful for clinic therapy of anti-tumor as a new regulator of PPARγ in the future.]]> 2008/12/9 0:00:00 YUAN Xian-Lin,HE Feng,CHEN Qing,YANG Xiang-Ling,YANG De-Po,WANG Dong-Mei and ZHONG Ling 832true 2009/1/10 0:00:00 WANG Chong,ZHONG Liang,WANG Dong-Sheng,LIU Bei-Zhong,LIAO Fei,HAO Po,LIU Chang,JIN Dan-Ting and WANG Chun-Guang 831true ezrin gene transcription are not well understood. Transcriptional regulation regions of the human ezrin gene were examined by linking 5′-deletion mutants and site-directed mutagenesis of the 5′-flanking region to a luciferase reporter gene, and the basal promoter region and key DNA sequence elements (an Sp1 binding site, -75/-69; and an AP-1 binding site, -64/-58) involved in the expression of the human ezrin gene in lung cancer A549 cells were explored. Furthermore, electrophoretic mobility shift assay (EMSA) showed that DIG-ddUTP labeled DNA sequences containing ezrin key elements could bind nuclear extracts from lung cancer cells to form DNA-protein complex, and the bindings to Sp1 and AP-1 sites by rhSp1 and rhAP-1, respectively, were specific. Finally, the analysis of transient transfection of A549 cells by transcription factor expression vectors indicated that Sp1 and AP-1 (c-Jun/c-Fos heterodimer) could increase the human ezrin basal transcriptional activity significantly through the Sp1 and AP-1 binding sites, respectively. In addition, over-expression of Sp1, c-Jun or c-Fos could up-regulate Ezrin expression. The data suggested that the Sp1 and AP-1 binding sites were two key elements regulating ezrin gene basal transcriptional activity in lung cancer cells, and the corresponding transcription factors Sp1 and AP-1 were crucial for the transactivation.]]> 2008/10/13 0:00:00 GAO Shu-Ying,LI En-Min,MENG Ling-Ying,CUI Lei,YUAN Hua-Min,DU Ze-Peng and XU Li-Yan 830true P < 0.01), compared with cells only induced by nerve growth factor and transfected by expression vector without CRMP-1. These data suggested that CRMP-1 play an important role in inhibiting neurite outgrowth.]]> 2008/9/15 0:00:00 GUO Guo-Qing,XIN Li,QIU Xiao-Zhong,SHEN Wei-Zai and YUAN Lin 829true 2008/9/22 0:00:00 JIA Jun-Ying,WANG Yun-Bo and TANG Jie 828true 18O/16O labeling method followed by Nano-HPLC-MS/MS were conducted to identify the differentially expressed proteins between gastric adenocaicinoma and nonmalignant gastric mucosa. A total of 78 differential proteins were identified, among these proteins, 42 proteins are over-expressed in gastric adenocarcinoma tissues and 36 proteins are down-expressed. Some of these differentially expressed proteins (moesin, periostin, annexin A2, annexin A4) were further confirmed by Western blot analysis. The quantitative proteomic protocol of 18O stable isotope labeling coupled with LCM was an effectively alternate technique for complicated proteins such as gastric cancer.]]> 2008/10/21 0:00:00 ZHANG Zhi－Qiang,LI Mao-Yu,ZHANG Gui-Ying,PENG Fang,YAO Hui-Xin,LI Mei-Xiang,XIAO Zhi-Qiang and CHEN Zhu-Chu 827true pcsk9 siRNA on THP-1 derived macrophages apoptosis induced by oxLDL，THP-1 derived macrophages were induced to differentiate into macrophages by PMA treatment for 24 h. The experiments were designed as follows: cells were incubated with oxLDL with a concentration of 0, 25, 50, 75, 100 mg/L for 48 h respectively. The apoptosis of THP-1 derived macrophages was observed by staining with Hoechst33258. The expression of pcsk9 was analyzed by reverse transcription polymerase chain reaction and Western blot. The siRNAs for pcsk9 gene were designed and synthesized，then transfected into THP-1 derived macrophages by positive ion liposome Lipofectamine 2000. Transfection efficiency was assessed by fluorescence microscope assay. RT-PCR and Western blot were conducted to detect the expression of pcsk9 after 24 h, 48 h respectively. The most efficient siRNA was selected to transfect into THP-1 derived macrophages. 24 h after transfection, cells were treated with oxLDL for 48 h, and then Hoechst 33258 staining. The results showed that the number of cells with nuclear condensation induced by 75 mg/L oxLDL increased significantly. In THP-1 derived macrophages, pcsk9 was upregulated with increasing concentration of oxLDL, while 75 mg/L oxLDL increased significantly. The RNA interference experiment showed that siRNA was successfully transfected into cells and 80 nmol/L as most effective dose of siRNA was selected by RT-PCR and Western blot. Compared with control, the suppression of apoptosis in THP-1 cells transfected with 80 nmol/L of siRNA for 24 h and incubated with 75 mg/L of oxLDL for another 48 h was detected by Hoechst 33258 staining and flow cytometer. Together, these results reveal the expression of pcsk9 mRNA and protein were increased by oxLDL in a concentration-dependent manner. Expression of pcsk9 gene could be effectively suppressed by siRNA. The apoptosis of THP-1 derived macrophages induced by oxLDL could be effectively suppressed by pcsk9 siRNA.]]> 2008/8/1 0:00:00 LIU Lu-Shan,XIE Min,JIANG Zhi-Sheng,YANG Qiong,PAN Li-Hong,WU Chun-Yan,TANG Zhi-Han,TANG Chao-Ke,WEI Dang-Heng and WANG Zuo 826true 2008/10/13 0:00:00 WANG Ge-Fei,PENG Cheng,LI Wei-Zhong,XIN Gang,SU Yun,CHEN You-Ying,LIN Gui-Mei and LI Kang-Sheng 825true SCN3A gene, 5′-Full RACE was performed to identify that the nucleotide “A” was identified as the transcription start site, which locate 27 kb upstream of the translation start site of human SCN3A and 31 kb upstream of the translation start site of mouse SCN3A. Two 5′ -untranslated exons of human SCN3A and one 5′-untranslated exon of mouse SCN3A were found by sequence blast. The core promoter region (-80 ～ +70) of human SCN3A showed 96% nucleotide homology with that of mouse. Two core promoter elements, BRE and TATA, were predicted in the region of -80～+70 from both human and mouse. The transcriptional factors PHR1, GATA-1, FOXN2, NF-1 and AP-4 predicted in the region of -400 to +200 of human SCN3A, not found in mouse SCN3A, and the transcriptional factors Sp、Sp3 and GBF predicted in the region of -400 to +200 of mouse SCN3A, not found in human SCN3A. The comparison between the promoter region of human and mouse SCN3A may provide an important clue to explore the mechanisms of the regulations of human and mouse SCN3A expression.]]> 2008/8/19 0:00:00 LONG Yue-Sheng,ZHAO Qi-Hua,ZENG Tao,ZENG Yang,SUN Wei-Wen and LIAO Wei-Ping 824true 1～3) were designed and chemically synthesized. The OGT-siRNA1～3 were transfected into HEK293T cells via lipofectamine2000. The efficacy of RNA interference was detected by RT-PCR. The fluorescence of GFP/OGT was counted by fluorescence microscopy after pEGFP/OGT and OGT-siRNA1～3 cotransfected to HEK293T cells for 24 h. The level of tau phosphorylation and glycosylation were detected by Western blot after Plasmid pCI/tau441 and the siRNA cotransfected HEK293T cells for 48 h. OGT-siRNA3(100 nmol/L) could effectively downregulate the expression of OGT gene compared with Mock group (80.0% at mRNA level and 51.3% at protein level). The level of phosphorylation at various sites of the tau was significantly upregulated and the glycosylation was downregulated following the inhibition of OGT gene expression. There is an apparent negative correlation between the modification of phosphorylation and glycosylation of tau protein, the deficient in glucose uptaken/metabolism may be an important pathogenesis of AD.]]> 2008/12/17 0:00:00 YANG Jiang-Yong,GU Jian-Lan,SHI Jian-Hua,LIU Fei and SHEN Qin 823true P < 0.05), and the expression of PIWIL4 mRNA in ovarian cancer tissues was higher than that in adjacent normal tissues (P < 0.01). MTT and clone formation analyses demonstrated that the cell growth activity and clone formation ratio of PIWIL4-siRNA transfected ES-2 cells were markedly decreased (P < 0.05). In conclusion, the expression of PIWIL4 mRNA was highest in ovarian cancer cell line ES-2 cells and was significantly increased in ovarian cancer tissues. Moreover, PIWIL4-siRNA can efficiently inhibit the cell activity and proliferation ability of ES-2 cells. In a word, PIWIL4 may be related to the pathogenesis of human ovarian cancer.]]> 2008/9/22 0:00:00 GUO Li-Min,LIU Min,LI Xin and TANG Hua 822true ca), temperature sensitive (ts), live attenuated influenza B virus strain B/Ann Arbor/1/66 was chosen for influenza virus rescue research, in which six internal gene segments, PB1, PB2, PA, NP, M, NS, were fully synthesized and nine amino acid substitutions were artificially alter by human intervention. The resultant B/Ann Arbor/1/66 plasmids were named as pAB121-PB1, pAB122-PB2, pAB123-PA, pAB124-HA, pAB125-NP, pAB126-NA, pAB127-M and pAB128-NS, respectively. A recombinant influenza A virus was previously generated entirely from cloned cDNA. An infectious recombinant influenza B virus was generated here, and designated as rMDV-B, by plasmid-based reverse genetics. The rMDV-B virus contained HA and NA genes from an epidemic influenza B virus strain B/Malaysia/2506/2004 in the background of internal genes derived from influenza B virus strain B/Ann Arbor/1/66. HA titer of rMDV-B in MDCK cells and embryonated chicken eggs ranged from 1∶64 to 1∶512. The results may allow an effective live influenza B vaccine to be produced from a single master strain, providing a model for the design of future live human influenza vaccines.]]> 2008/10/29 0:00:00 YANG Peng-Hui,AN Wen-Qi,SHI Xin-Fu,DUAN Yue-Qiang,LUO De-Yan,ZHANG Peng-Fei,TANG Chong,XING Li,ZHANG Yu-Jing,LIU Xiu-Fan and WANG Xi-Liang 821true Dlc8) gene expression pattern in different gravitational environments. The open reading frame sequence of Dlc8 gene was amplified and analyzed. The distribution of Dlc8 gene was investigated in embryo, head, silk gland, midgut, cuticle, blood, fat, tuba malpighii of silkworm respectively. The effects of simulated gravitational environments (0 g, 1 g, 2 g) produced by large gradient high magnetic field (LGHMF) on expression of the silkworm Dlc8 gene were tested by RT-PCR and real time RT-PCR in BmN cells. Expression of the silkworm Dlc8 gene in simulated weightless environment was tested by real time RT-PCR in inverse period and in total embryo period of silkworm. The Dlc8 gene with 270 bp coding 323 aa was successfully amplified. The homology rates of amino acid sequences of Dlc8 gene between silkworm and Arabidopsis thaliana, Drosophila melanogaster, Caenorhabditis elegans, Xenopus tropicalis, Mus musculus, Homo sapiens were 67%, 96%, 91%, 95%, 92%, 92%, respectively. Signal peptide analysis showed that Dlc8 was not a secretion protein. There was not glycosyl-phosphatidyl inositol anchor site in Dlc8 amino acid sequence. Molecular mass and isoelectric point of Dlc8 were 10.34 ku and 6.81, respectively. Dlc8 gene is conservative in embryo, head, silk gland, midgut, cuticle, blood, fat, tuba malpighii of silkworm. Dlc8 gene is more sensitive to gravity than magnetic field in BmN cells. There are different responses to gravity on Dlc8 gene expression in different period of silkworm embryo. There was no significant change of Dlc8 gene expression between simulated weightless and control groups. The results showed that Dlc8 gene could be the molecular target to study gravity bioeffect. This research may contribute to reveal the mechanism of gravity bioeffect of the silkworm Dlc8 gene.]]> 2008/10/29 0:00:00 TIAN Zong-Cheng,QIAN Ai-Rong,XU Hui-Yun,DI Sheng-Meng,ZHANG Wei,WENG Yuan-Yuan and SHANG Peng 820true 2008/12/17 0:00:00 DAI Jin-Xiang,CHEN Keng,JIN Wei-Lin and JU Gong 819true 2008/11/18 0:00:00 LIU Jing and WANG Yan-Ling 818true 2008/8/19 0:00:00 JIANG Yuan-Yuan,LIU Ming-Yao,REN Gui-Ping,WANG Wen-Fei,LIU Xiao-Min and LI De-Shan 817true 3. The FGF-21/receptor complexes were immunoprecipitated by either Flag antibody or streptavidin and analyzed by SDS-PAGE. The results showed that FGF-21 could form putative FGF-21/receptor(s) complexes with surface membrane protein of differentiated 3T3L1 adipocytes, not pre-adipocytes. The molecular mass of the complexes is proximate 300～400 ku. The complexes were formed by the FGF-21 labeled with either Flag-tag or biotin, suggesting the complex formation was not influenced by different labeling. To understand the component of the FGF-21/receptor(s) complexes, Western blot assay was used to examine the interaction of the complexes with known antibodies. FGFR-2 was detected within the FGF-21/receptor complexes by specific antibody for FGFR-2. To confirm the specific relationship between FGF-21 and FGFR-2, FGF-21-induced FGFR-2 tyrosine-phosphorylation was studied in 3T3L1 adipocytes. The results showed that although FGFR-2 was expressed in both pre-adipocytes and adipocytes FGF-21 could only enable adipocyte-expressed FGFR-2 tyrosine phosphorylation, which was consistent with glucose-uptake by FGF-21 in this two cell types. FGF-21 not only phosphorylated in situ adipocyte-expressed FGFR-2, but also ectopically expressed FGFR-2 in mouse pre-B cell line BaF3 cells. Sequencing analysis of FGFR-2 cDNA from adipocytes revealed that FGFR-2Ⅲc was the only subtype of FGFR-2 expressed in adipocytes, suggesting that FGFR-2Ⅲc is at least one of the functional receptors for FGF-21 and involved in glucose metabolism in the cells. In order to understand the reason why the same FGFR-2 only functions in adipocyte and not in pre-adipocytes, differential expression of FGFR-2 before and after 3T3L1 cell differentiation was systematically analyzed, the result showed that adipocytes expressed high level precursors of FGFR-2, which were not observed in pre-adipocytes, suggesting FGFR-2 turn-over rate could be a key factor to determine the function the molecule. Alternatively, adipocyte-specific molecules could be another key factor participating the signal transduction of FGF-21.]]> 2008/8/20 0:00:00 REN Gui-Ping,LI Lu,SUN Guo-Peng,HOU Yu-Ting,WANG Wen-Fei and LI De-Shan 816true via TCF-4 pathway. To further study mechanism of spindlin1 function, based on the bioinformatics analysis of spindlin1 structure, A series of spindlin1wild or mutant expression vectors was constructed to determine the key amino acid points for spindin1 localization and function. As previously reported, wild type spindlin1 protein was localized in the nuclei of HeLa cells. Cells transfected with construct either with mutation of Ser14＋Ser84, Ser84＋Ser99 or Ser14＋Ser84＋Ser99 exhibited a cytoplasm spindlin1 expression in a diffused manner, while in those cells transfected with construct with mutation of Ser14，Ser84, Ser99 or Ser14＋Ser 99, the spinlin1 showed a similar subcellular nuclear location as wild type spindlin1. Further TCF-4 reporter assay were performed using these wild type or mutant spindlin1 constructs, and the results indicated that mutation of Ser84 with either Ser14 or Ser99 could abolish the promotion of spindlin1 on TCF-4 reporter activity. All these result suggested that Ser84 of spindlin1, with the cooperation of Ser14 or Ser99 play a key role in it nuclear localization and its function in regulation of TCF-4 pathway.]]> 2008/10/29 0:00:00 CONG Bin,ZHANG Peng,WANG Jing-Xue,ZENG Quan,CHEN Lin,YUE Wen and PEI XUE-Tao 815true Kcat/Kuncat was about 1.6×104. The catalytic activity of the catalytic antibody in a reverse micelle reaction system based on sodium bis (2-ethylhexyl) sodium sulfosuccinate (AOT) in isooctane was studied. Kinetic analysis of the catalytic antibody-catalyzed reaction was found to be possible in this system. Kinetic studies showed that hydrolysis in the microemulsion system follow Michaelis-Menten kinetics. The catalytic antibody can also accelerate catalysis of S-ibuprofen methyl ester in the microemulsion system. Temperature effects, the pH profile, Km,app and Kcat were determined. The dependence of the catalytic antibody hydrolytic activity on the wo (molar ratio of water to surfactant) showed a bell-shaped curve, presenting a maximum at about wo = 21.]]> 2008/8/21 0:00:00 YANG Gen-Sheng,QI Ying-Dan,OU Zhi-Min and YAO Shan-Jing 814true 2008/8/20 0:00:00 QU Hao,ZHANG Tao,ZHANG Li-Yan,ZHANG Yi,WANG Mei,WANG Dong-Mei,HE Da-Shui,HUANG Li-Hua,ZHU Wei-Bin and ZHANG Yu-Guang 813true +-dependent glycerol 3-phosphate dehydrogenase (GPD) are rate limiting for glycerol production in Saccharomyces cerevisiae. Recently, the gene CgGPD encoding glycerol 3-phosphate dehydrogenase homologous to GPD genes in other yeasts was cloned from Candida glycerinogenes WL2002-5, an excellent industrial glycerol producer. However, the knowledge about CgGPD expression regulation, especially difference with GPD1 and GPD2 from S. cerevisiae is so less. A functional comparison of CgGPD from C. glycerinogenes with GPD1 and GPD2 from S. cerevisiae was undertaken, using S. cerevisiae gpd1/gpd2 and gpd1 osmosensitive mutants as expression systems. The functions of three indicated genes in S. cerevisiae was characterized for osmoregulation under high osmotic stress and redox regulation under anaerobic condition with various transformants. The results showed that gpd1/gpd2 mutants harbouring CgGPD and GPD1 can restore osmotolerance and increase glycerol production ability under hyperosmotic stress but mutant expressed GPD2 can not. When cells were cultured under the anaerobic condition, the growth pattern of mutants harbouring CgGPD and GPD2 are smilar, however the mutant harbouring GPD1 grows slowly and the growth of the controll supresses thoroughly. Furthermore, gpd1/gpd2 mutant employing either CgGPD or GPD2 can increase glycerol production ability and improve GPD specific enzyme activity in anaerobic incubation, but gpd1/gpd2 mutant expressed GPD1 has no the similar results under the same condition. This indicated that CgGPD may involve in both osmoregulation and redox balance and can complement GPD1 and GPD2 in gpd1/gpd2 mutant.]]> 2008/9/22 0:00:00 CHEN Xian-Zhong,FANG Hui-Ying,RAO Zhi-Ming,SHEN Wei,ZHUGE Bin,WANG Zheng-Xiang and ZHUGE Jian 812true Salmonella enterica serovar Typhi, a gram-negative human enteroinvasive pathogen is the etiological agent of typhoid fever, and is an important experimental model for prokaryote research. DNA microarray technology was widely used in analysis of genomic structures and expression profiles. The genomic DNA microarrays were prepared based on the genomic sequences of S. enterica serovar Typhi. A total number of 4 201 protein encoding genes selected from strains Ty2, LT18 and a z66+ wild strain of S. enterica serovar Typhi were amplified by PCR. The products were purified and printed onto the poly-L-lysine coated chip slides in duplicate to form the genomic DNA microarrays. Fluorescently labeled probes were prepared by priming of genomic DNAs with random hexamers and extension with Klenow DNA polymerase. Labeled DNAs were hybridized with the microarrays to check the printing effect and verify the genes order. The genomic DNA microarray-based bacterial gene expression profiling analysis was optimized and used to investigate global transcriptional responses when wild strain of S. enterica serovar Typhi was grown in high- and low-osmotic environmental conditions. The major results were consistent with those of previous research with oligo-genomic microarrays of S. enterica serovar Typhi. These results demonstrated that S. enterica serovar Typhi genomic DNA microarrays were successfully prepared and could be utilized in relative gene expression profiling analysis and comparative genomic researches.]]> 2008/8/1 0:00:00 SHENG Xiu-Mei,HUANG Xin-Xiang,MAO Ling-Xiang,ZHU Chao-Wang,XU Shun-Gao,ZHANG Hai-Fang,XU Hua-Xi and LIU Xiu-Mei 811true 2008/7/14 0:00:00 ZHANG Qiang and ZHAO Xin-Huai 810true 2008/10/29 0:00:00 WANG Ben-Xu,LIU Yu,LIU Zhan,GAO Ya-Ping,WANG Fang,PAN He-Ping,YANG Guang,XU Hua,SHEN Bei-Fen,LIU Chuan and SHAO Ning-Sheng 809true P < 0.05). These data indicate that 5-AZA-CdR inhibits the development of mouse preimplantation embryos. Apoptosis, DNA methylation, and transcriptional activity were analyzed to determine the reason for these developmental defects. An annexin V-PI assay revealed that high doses of 5-AZA-CdR led to apoptosis. Compared to the controls, DNA methylation was significantly reduced in uncompacted 8-cell embryos and morulae ( P < 0.05) in a dose- dependent manner, whereas no significant change was detected in 2- or 4-cell embryos ( P > 0.05). The observed changes in transcriptional activity, determined by measuring the incorporation of BrUTP, were similar to the observed alterations in DNA methylation. Therefore, the developmental defects induced by 5-AZA-CdR appear to be mediated by alterations in DNA methylation and transcriptional activity in preimplantation mouse embryos.]]> 2008/11/18 0:00:00 YU Jian-Ning,LI Shao-Hua,WANG Dan-Qiu,WANG Meng,LIN Fei and LIU Hong-Lin 808true 2008/11/4 0:00:00 TIAN Hui,XU Jie,LIU Xian-Xi,ZHANG Bing,LI Wen-Jun and SONG Xu 807true 2008/7/7 0:00:00 SUN Yi-Min,ZENG Ling-Qin,ZHANG Yan,WEI Li,ZHANG Liang,CHENG Jing and K R MITCHELSON 806true 2008/8/20 0:00:00 WU Li Hui,HU Ping,WU Wei and WU Xing Zhong 805true 2008/6/16 0:00:00 FU Yu-Rong,YI Zheng-Jun,lI Meng,SHAO Li-Jun and SONG Shu-Ya 804true trans-Golgi network (TGN) 38, markers for the cis- and trans-Golgi apparatus, respectively. Morphometric analyses indicated that ～60% of the ppGalNAcT2 signal colocalized with the GS28, while ～36% of the cis-Golgi marker colocalized with the ppGalNAcT2. Approximately 34% of the ppGalNAcT2 signal colocalized with the TGN38, whereas 38% of the trans-Golgi marker colocalized with the ppGalNAcT2. The results provide unequivocal evidence for the location of ppGalNAcT2 within the Golgi apparatus, and further highlight the importance of this organelle in the initiation of O-linked glycosylation.]]> 2008/8/1 0:00:00 ZHOU Ying-Hui,HANG Sai-Yu,QIU Hao,JIA Wei,XU Lan,JIANG Zhi and WU Shi-Liang 803true mdr1 gene in the KB/TAX cells were remarkable reduced. Moreover, both KB and KB/TAX cells were sensitive to STA-35, the relative resistance to TAX on KB/TAX cells was decreased by the addition of STA-35. Furthermore, STA-35(5～20 μmol/L)was capable to reduced the activity of P-gp by increasing the accumulation of rhodamine 123, decreasing P-gp expression in KB/TAX cells in a dose dependent manner, but had no effect on the mdr1 gene expression. These results suggest a potential action of STA-35 as MDR reversing agent, and one of the possible mechanisms could be the suppression of P-gp function and protein expression.]]> 2008/9/16 0:00:00 YI Wen-Yuan,XU Bo,LI Min,LI Zhong-jun and CUI Jing-rong 802true 2008/9/22 0:00:00 LIU Chun-Guo,LIU Ming,ZHANG Yun,LIU Da-Fei,PAN Wei-Qi,SUN En-Cheng,DU Jin-Ling and LI Hong-Tao 801true 2008/11/3 0:00:00 Ding Dao-Fang,XING San-Li,ZHOU Ming-Ming and SOU Hou-Yan 800true 2O2 and NaCl, but weakly influenced by several plant hormones. Time- and dose-dependent effects were observed in both H2O2 and NaCl treatments, and the strongest induction was observed when rice seedlings were treated with 0.5 mmol/L H2O2 for 12 h or 500 mmol/L NaCl for 24 h. Additionally, dimethylthiourea, a H2O2 trap, inhibited H2O2-enhanced expression of OsPHGPx, but did not impair the enhanced effect of NaCl, implying that NaCl-induced OsPHGPx expression was not mediated by H2O2. These results will contribute greatly to further study the exact physiological function of OsPHGPx in rice.]]> 2008/9/11 0:00:00 Li Tian,YANG Xiao-Dong and Liu Jin Yuan 799true in vitro and the changing of cellular cytoskeleton induced by PMA was investigated with atomic force microscopy. PMA is usually used to stimulate PKC signal transduction pathway in synoviocytes and to simulate process of inflammation. The results presented here show that synoviocyte cytoskeleton has changed significantly after PKC activation by using the imaging of atomic force microscopy. The results also indicate that the activation of PKC might play crucial role in cytoskeleton changing of synoviocytes，which, to some extent, provides some experiments and ideas on understanding the mechanism underlying rheumatoid arthritis (RA).]]> 2008/8/19 0:00:00 PAN Lei-Ting,SUN Wen-Wu,QI Ji-Wei,WU Xian,SONG Kun,HU Fen,ZHANG Xin-Zheng and XU Jing-Jun 798true D10Wsu52e gene, is a member of function-unknown protein family, UPF0027. These protein family homologues are conserved and ubiquitously expressed in various species, organs and cells. But the functions are quite unclear yet. Here, FAAP were demonstrated to deposited in the cytoplasmic fractions and the plasma membrane fractions by ultra-speed centrifugation. The expression levels of FAAP are significantly correlative with 67 ku Laminin receptor (LR) in HeLa cells. Moreover, FAAP, similar with LR, could effectively restrain the cultured cells attachment. Together, these results suggest the effects of FAAP on cell attachment.]]> 2008/6/19 0:00:00 HU Jin－Song,DING Nai-Zheng,TENG Jun-Lin and CHEN Jian-Guo 797true in vitro. mESC-D3, maintained in adherent monolayer culture condition, were induced to differentiate along hepatic lineage with addition of some factors to the medium at different time. The factors included FGF, HGF, OSM and so on. The differentiated cells showed a hepatocyte-like morphology, expressed hepatic marker genes and have also produced and stored glycogen by phase contrast and transmission electron microscopy, reverse transcription-polymerase chain reaction, immunocytochemistry, and periodic acid-Shiff staining respectively. Stem cell differentiation-related microarray was used to analyze the differential gene expression profiling during hepatic differentiation of mESC-D3 at early stage. Quantitative PCR was performed to verify the microarray data. Microarray analysis presented 48 genes expressed differentially (2 fold), including 20 genes up-regulated and 28 genes down-regulated. Further bioinformatics analysis showed the majority of these genes were extracellular matrix, intercellular junction and FGF, BMP, Notch and Wnt signaling pathways molecules, which suggests these alterations may be closely associated with the hepatic differentiation of embryonic stem cells at early stage.]]> 2008/9/16 0:00:00 LI Yan,LI Xue-Fei,GUO Kun,GAO Dong-Mei,ZHAO Yan and LIU Yin-Kun 796true 2008/10/29 0:00:00 ZHAO Cheng-Jun,DENG Qi-Yue,ZHANG Dong-Mei,CAI Wen-Qin and Zhang Ji-Qiang 795true ca gene was divided into four overlapping fragments and expressed in E. coli BL21 respectively. Pepscan technology was employed to screen the antigen epitope through the expressed fusion proteins were detected respectively with three McAbs by Western blot. Then three liner epitopes recognized by three McAbs were preliminary identified, and the functional affinities of anti-CA McAbs were assessed with non-competitive ELISA method. All this may be helpful in understanding molecular properties of JSRV-CA and may be useful for pathogenic diagnosis and vaccine design.]]> 2008/10/29 0:00:00 LUO Jun-Rong,Siriguleng,LIU Xiao-hui,ZHOU Jian-hua and MA Xue-en 794true in vitro, Ad-HSP70p-E1A can replicate in lung carcinoma A549 cell line and destroy those cancer cells to some extent. While combined with hyperthermia, the number of replicative virions released from lung carcinoma increased about 2～10 times and the oncolytic effect on lung carcinoma A549 cell line increased 5 times at least. In addition, when combined with oncolytic adenovirus Ad-HSP70p-E1A, replication-deficient adenoviruses such as AdGFP, Ad-CMV-hGMCSF and Ad-CMV-mIL12 at the moi of 10 are expected to lead to the expression level of therapeutic gene at higher levels. The green fluorescence protein increased 76.64 times, while cytokines GMCSF and IL12 increased 5 times and 7 times respectively.]]> 2009/8/7 0:00:00 WEI Fang,WANG Hui-Ping,CHEN Xia-Fang,LI Chuan-Yuan and HUANG Qian 793true Saccharomyces cerevisiae, which was supposed to control pleiotropic drug resistance of cell and probably regulate genes involved in stress response. It still lacks enough evidence to conclude that Rdr1 involves in stress response. RDR1 gene was cloned by PCR and ligated into multicopy vector pYES2/NTA resulting an overexpression plasmid pYES2/NTA-RDR1. The overexpression of Rdr1 protein was induced by 2% galactose to achieve a gain-of-function phenotype. In order to reveal the function of transcription repressor Rdr1 in stress response, the growth state of wild type cells, cells with overexpressed Rdr1 protein and RDR1 mutant cells under H2O2, heat-shock and salt stress treatments were compared. The data demonstrated that cells with overexpressed Rdr1 protein were hypersensitive to above three stress treatments, while RDR1 mutant cells were equal or more tolerant to theses stress treatments compared to wild type cells. Yeast cells elevate antioxidant enzymes activity when challenged with different stressors. The role of RDR1 in stress response may relate to its regulation of antioxidant enzymes activity. To verify this hypothesis, the activities of antioxidant enzymes including superoxide dismutase (SOD), catalase, glucose-6-phosphate dehydrogenase (G6PDH), glutathione reductase (GR) were measured. It was showed that RDR1 mutant cells had higher activities of SOD, catalase, G6PDH and GR compared to wild type cells, while cells with overexpressed Rdr1 protein had lower activities of SOD, catalase, G6PDH and GR. The negative effect of RDR1 on activities of SOD and catalase was more intense than it on activities of G6PDH and GR. The results might explain the phenomenon that overexpressed Rdr1 protein were hypersensitive to above three stress treatments, while RDR1 mutant cells were equal or more tolerant to theses stress treatments compared to wild type cells. Therefore, the study provided preliminary genetics and biochemistry evidences for the negative regulatory role of transcription repressor Rdr1 in stress response.]]> 2009/8/25 0:00:00 MIAO Min,CAO Hong-Ping,ZHONG Yan,LIU Jun,WANG Yi-Hui,LIU Xin,ZHANG Nian-Hui and LIU Ke 792true in vitro tissue engineering models. It was hypothesized that human insulin-like growth factor-1(IGF-1) and three dimensional dynamic microenvironment could act independently and interactively to enhance the survival and differentiation of adipose tissue-derived stem cells (ADSCs) and hence the construction of engineered cardiac grafts. IGF-1 can be expressed by the ADSCs through genetic modification, which can be conveniently realized by incorporating the relevant genes into the three dimensional scaffold. ADSCs were cultured on three dimensional porous scaffolds with or without plasmid DNA PIRES2-IGF-1 in cardiac media, in dishes and in a spinning flask bioreactor respectively. Cell viability, formation of cardiac like structure, expression of functional proteins, and gene expressions were testified to the cultured constructs on day 14. The results showed that dynamic microenvironment enhanced the release of plasmid DNA; the ADSCs can be transfected by the released plasmid DNA PIRES2-IGF-1 in scaffold; IGF-1 had beneficial effects on the cellular viability and the increase of total protein; and it also increased the expressions of cardiac specific proteins and genes in the grafts. It was also demonstrated that dynamic stirring environment could promote the proliferation of ADSCs. Therefore, IGF-1, expressed by ADSCs transfected by DNA PIRES2-IGF-1 incorporated into scaffold, and hydrodynamic microenvironment can independently and interactively increase cellular viability, and interactively increased the expressions of cardiac specific proteins and genes in the grafts. The results would be useful for developing tissue engineered grafts for myocardial repair.]]> 2009/7/27 0:00:00 ZHU Yan-Xia,LIU Tian-Qing,SONG Ke-Dong,MA Xue-Hu and CUI Zhan-Feng 791true ATP channel current (IKATP) was monitored using the whole cell configuration of the patch clamp technique in single ventricular cardiac myocytes enzymatically isolated from rat hearts. Sodium cyanide, a metabolic inhibitor, was used to mimic cell ischemia. Morphine postconditioning facilitated the further opening of the KATP channels. Following initial activation of IKATP by mimic ischemia, morphine further increased current amplitude by (61.4±13.6)%. KATP channels in normal cardiac myocytes were not affected by extracellular applications of morphine alone. Nor-binaltorphimine, a specific κ-opioid receptor antagonist, was unable to abolish the facilitation of morphine postconditioning when administered 5 minutes before morphine. However, naloxone, a non-specific opioid receptor antagonist, and naltrindole, a specific δ-opioid receptor antagonist, can abolish the facilitation of morphine postconditioning. These results indicated that morphine postconditioning facilitate the further opening of the cardiac sarcolemmal KATP channels following the channel activation by mimic ischemia. The δ-opioid receptor may be involved in morphine postconditioning.]]> 2009/7/15 0:00:00 TANG Zhen-Shen,NIU Wei-Zhen,WEI Hua,LI Tian-Zuo,LIU Ping and ZHANG Bing-Xi 790true Rattus norvegicus (Cav22P for short) was amplified by high fidelity PCR, inserted into vectors pET28b and pGEX-4T-1 respectively, and expressed in Escherichia coli Rosetta. The expressed product HIS-Cav22P mostly deposited in an inclusive body. The inclusive body of HIS-Cav22P dissolved in urea buffer. After dilution and dialysis, the refolded protein HIS-Cav22P was finally purified via Histrap chelating HP column. Ultraviolet spectroscopy results demonstrated that HIS-Cav22P protein can bind to calcium ions reversely. The expressed product GST-Cav22P was purified from the supernatant of cell lysate using Glutathione Sepharose 4B column. However, GST-Cav22P degraded severely, which caused it difficult to purify the protein Cav22P and the GST pull-down assay. All the results suggest that the active recombinant proteins of Cav22P may act as a molecular target for high through screen of non-narcotic analgesic drugs.]]> 2009/9/7 0:00:00 ZHOU Yu-Juan,XIE Li-Ping and ZHANG Rong-Qing 789true in vitro and its NO and ERK1/2 pathway, perfusion method in vitro vascular ring and Power-Lab system was used to detect tension on rat’s vascular rings. Experimental groups below: Phenylephrine group, Acetylcholine group, apelin-13 group, apelin-13+ PE group, apelin-13+Ach group, PD98059+PE group, PD98059+Ach group, LNNA+PE group, LNNA+Ach group, apelin-13(preincubation)+PD98059+PE group, apelin(preincubation)-13+PD98059+Ach group, apelin-13(preincubation)+LNNA+PE group and apelin-13 (preincubation)+LNNA+Ach group, compared with WKY rats. Rats’ vascular smooth muscle cells were cultured and the expression of ERK1/2 protein was detected by Western blot. Apelin-13 for the blood vessel with endothelium has demonstrated concentration-dependent vasodilation and SHR < WKY in percentage vasodilation, but for the blood vessel without endothelium, apelin-13 shows the role of vascular contraction and SHR > WKY in contraction tension. Apelin-13 pre-incubation can reduce the SHR and WKY rats’ the reactivity of vascular contraction tension on phenylephrine and increases the relaxation response to acetylcholine. After NOS inhibitors LNNA blocking the formation of NO, the relaxation response of the vascular rings to apelin-13 is significantly inhibited and apelin more pronounced reduces the diastolic response in the SHR group than in the WKY group. This suggests that the vasodilator effect of apelin-13 partly depends on NO-dependent pathway at least and SHR hypertensive rats with NO pathway obstacles reduces the vasodilation of blood vessels to apelin-13. After pre-incubation of ERK1/2 inhibitor PD98059, the response of vascular rings to apelin-13 shows concentration-dependent contraction, which is the same as the blood vessel without endothelium to apelin-13, SHR > WKY in contraction tension. PD98059 reversals the apelin-13’s vasodilation effect. Western blot analysis showed that apelin-13 promoted the concentration-dependent and time-dependentexpression of pERK1/2. The potent ERK inhibitor PD98059 decreased the expression of pERK1/2. The diastolic reactivity of apelin in ex-vivo vascular rings of SHR is reduced and the effect is mediated by NO pathway and the ERK1/2 pathway.]]> 2009/7/27 0:00:00 LIU Chang-Hui,LI Xin,CHEN Feng,PAN Wei-Nan,FENG Fen,QIN Xu-Ping,LI Lan-Fang,SU Tao and CHEN Lin-Xi 788true in vitro．Accordingly, CREG might have potential applicational perspective in attenuating the progression of atherosclerotic plaques and restenosis after percutaneous coronary intervention．]]> 2009/10/15 0:00:00 WU Guang-Zhe,YAN Cheng-Hui,HAN Ya-Ling,TAO Jie,DENG Jie,TIAN Xiao-Xiang,ZHANG Bao-Hai,WANG Tao,KANG Jian and ZHANG Xiao-Lin 787true in vitro, three of the four miRNAs, miR-18a, miR-145 and miR-495, were proved to effectively inhibit Ptf1a expression, respectively, by binding to the 3′UTR of mouse Ptf1a mRNAs. Finally, expression patterns of miR-18a, miR-145 and miR-495 and Ptf1a were detected in the mouse pancreatic development via qRT-PCR and immunofluorescent staining. The results showed that the expression of miR-18a, miR-145 or miR- 495 is negatively correlated with that of Ptf1a mRNA and protein. Accordingly, these results indicated that miR-18a, miR-145 and miR-495 may regulate the expression of Ptf1a in developing mouse pancreas.]]> 2009/9/27 0:00:00 AN Yang,YANG Yan-Kun,GAO Fei,ZHU Kuan-Yu,MU Tan-Wei and TENG Chun-Bo 786true 2009/7/9 0:00:00 LIU Yu-Ming,PENG Zhi,DENG Wei-Wei,SHI Tai-Ping and MA Da-Long 785true 2009/7/9 0:00:00 XUE Jin-Feng,LIU Xiong-Hao,HE Qiang,XUE Zhi-Gang,HU You-Jin,LI Zhuo,YANG Jun-Ling,GAO Ting,PAN Qian,LONG Zhi-Gao,WU Ling-Qian,XIA Kun,LIANG De-Sheng and XIA Jia-Hui 784true presenilin-1 and presenilin-2 double knockout mice (dKO mice) exhibit several neurodegenerative phenotypes of Alzheimer’s disease (AD) pathology, such as tau hyperphosphorylation, neuron loss, forebrain cortical shrinkage and memory impairment. By using capillary electrophoresis assay, monoamine neurotransmitters in forebrain cortex, hippocampus and other forebrain region of dKO mice aged at 6, 9 and 12 months were measured to illustrate the relationship among presenilins function deficiency, neurodegenerative phenotypes and monoamine neurotransmitters. Data showed that levels of monoamine neurotransmitters in forebrain cortex of dKO mice were significantly decreased at 6 months when compared to controls, while as mice getting older, levels of monoamine neurotransmitters increased to that of controls, or even higher. In hippocampus, 5-hydroxytryptamin and epinephrine in dKO mice had a significant increase at 6 months, followed with a significant increase of each monoamine neurotransmitter at 12 months age. In other forebrain region, 5-hydroxytryptamin and dopamine had a similar level between control and dKO mice at 6 and 9 months but a significant decrease at 12 months; however, level of norepinephrine and epinephrine were significantly decreased at 6 and 12 months except epinephrine of 6 months. These results demonstrated that knockout of presenilins genes could lead to the variation of monoamine neurotransmitters, and the variation profiles were different among forebrain cortex, hippocampus and other forebrain region. However, whether presenilins deficiency caused the variation of monoamine neurotransmitter directly or not, and how about the effects of variation of monoamine neurotransmitters on AD-like pathology need to be further analyzed.]]> 2009/8/25 0:00:00 ZHANG Dong-Li,LIANG Li-Yun,JI Ting-Ting,CHEN Yi-Qun,ZHOU Tian-Shu and MEI Bing 783true 2009/6/26 0:00:00 CHEN Rong,QIU Zhu-Ping,ZHANG Yang and ZHOU Yi-Feng 782true 24～Met41 in N terminal of TLR4. Aggregation of TLR4 to LPS stimulation was observed, however, TLR4 without the Glu24～Met41 mutation did not aggregate. All these results indicated that TLR4 Glu24～Met41 might be the interaction domain of TLR4 binding to MD-2 and participate in the aggregation effect of TLR4 upon LPS stimulation.]]> 2009/6/30 0:00:00 ZHONG Tian-Yu,TANG Jing,CHEN Deng-Yu,LIU Ya-Wei,WANG Wei,LIU Jing-Hua and JIANG Yong 781true 2009/9/7 0:00:00 XIONG Xia,SHEN Jian-Ying,LI Jian-Jun,ZHANG Hai,WANG Xian-Chun and LIANG Song-Ping 780true in situ hybridization with BHC80 RNA probe showed that BHC80 was expressed in zebrafish central nervous system. Morpholino-modified antisense oligonucleotide was injected into zebrafish zygotes to knock down BHC80 expression. BHC80 knockdown resulted in striking decrease of erythrocytes and accumulation of erythrocytes at PBI. Further investigation of embryonic erythrocytes marker βe3 globin and hematopoiesis transcription factors gata1, c-myb and lmo2 by in situ hybridization showed that the erythroid progenitors marked with gata1 in BHC80 knockdown embryos were high proliferation and their differentiation were delayed, which led to decrease of erythrocytes and accumulation of erythrocytes at PBI. Both in situ hybridization and microangiography indicated that vasculature pattern of BHC80 knockdown embryos were almost normal.]]> 2009/8/7 0:00:00 HOU Jia-Yun,JIANG Qiu,ZHANG Jie,TAN Li and SONG Hou-Yan 779true ABCE1, siRNA-expressing vectors targeting sites of the ABCE1 gene were constructed from RNAi-Ready pSIREN-DNR-DsRed-Express vector. Cultured 95-D and NCI-H446 lung carcinoma cells were transfected with the siRNA-expressing vectors using FuGENE 6 and transfection efficiency was determined by using fluorescence microscopy. The expression level of ABCE1 protein was determined by Western blot and immunofluorescence staining. Cell viability was determined by MTT, cell cycle was analysed by flow cytometry.The apoptotic rate was observed by ELISA. Fluorescence microscopy showed a satisfactory transfection efficiency which was about 42.70%. Cell viability and the growth fraction were markedly suppressed,whereas the apoptosis was significantly increased in SiRNA-95-D and SiRNA-NCI-H446 cells than controls(P < 0.05). It can be concluded that the siRNA targeting ABCE1 gene shows a dramatic inhibitory effect on RNA transcription and protein expression and a promoting effect on the apoptosis in 95-D/NCI-H446 cells, which offers a reliable base for the further in vivo experiment.]]> 2009/8/7 0:00:00 ZHENG Mao-Gen,GAO Ying,HUANG Bo,TIAN Da-Li and YANG Chun-Lu 778true +(S. aureus ATCC2592 and Streptococcu ATCC55121), G-(E. coli ATCC25922) and fungi(C. albicans ATCC2002). The peptides did not show active hemolytic activity against rabbit blood red cells and had no significant effects on human blood coagulation system. The discovery of antibacterial peptides in sheep reproductive system reveals that antibacterial peptides may play a role in innate immunity against microorganisms in a wide range of animal species.]]> 2009/2/10 0:00:00 CHEN Chen,WANG Xin-Hua and BO Xin-Wen 777true 2009/7/21 0:00:00 SHI Yan,LIU Xiong-Hao,LIANG De-Sheng,FENG Mai,WU Ling-Qian,YANG Jun-Lin,LI Zhuo,ZHAO Kai,PAN Qian,LONG Zhi-Gao and XIA Jia-Hui 776true 2009/7/15 0:00:00 ZHANG Yan,WEN Wei and LUO Jin-Yong 775true 2009/5/27 0:00:00 XIAO Zhi-Ke,ZHANG Meng-Xia,JIANG Hao,WU You-Hua,AI Xiao-Hong,LUO Hong-Mei,WANG Yu-Hua,LEI Xiao-Yong and TANG Sheng-Song 774true Bacillus anthracis and it is a binary bacterium toxin composed of protective antigen (PA), lethal factor (LF) and edema toxin (EF). Among of them, LF is the enzyme moiety lead to the death of the cell. To investigate key function amino acid of LF and study the lethal mechanism of lethal factor, random mutagenesis was carried out. The mutation library of the 774 bp fragment of lethal factor from site 1 550 to 2 324 bp was constructed using DiversifyTM PCR Random Mutagenesis Kit (Clontech) with some modifiication. Transformants of lef mutation library were induced by 0.2 mmol/L IPTG at 28℃ for 4 h and then T7 phage were added into the culture, incubating for further 2～3 h to lyse the bacterium. The lysis was centrifuged at 12 000 r/min for 8 min and the supernatant which contained the target protein was collected. To determine the activity of the mutants, the cell cytotoxicity assay was carried out. RAW 264.7 microphage cells were seeded into 96-well plates, 24 h ahead of experiments. When assaying, the medium was removed and 100 μl DMEM medium containing 1.0 mg/L of purified recombinant and 10 μl of the lysis supernatant was added into each well. Then the plates were incubated at 37℃ for 4 h. The cell viability was determined by using the alamarBlueTM (AbD Serotec) dye according to the manufacturer’s direction. The mutants whose activities had decreased significantly were sequenced and further purified by Ni-histidine tag affinity chromatography. The cell cytotoxicity and competition binding assay were demonstrated to analyse the character of the mutants. Five mutants which had low biological activity determined through the cell cytotoxicity assay were obtained. Furthermore, two mutant proteins with single-site mutation, K518E and L519C, were purified for the first time. The cytotoxicity assay showed that the two mutant proteins lost activity significantly, that means, Lys518 and Leu519 played an important role in the function of lethal factor. The competition binding assay showed K518E endowed with the character of competition inhibition of wild type LF, whereas L519C doesn’t have the character. Probably, mutant K518E change Lys into Glu carrying negative charge and affects the catalytic groove of LF which is negative charge. Therefore, Lys 518 maybe has an important role to stabilize the conformation of LF and combine the substrate.]]> 2009/6/19 0:00:00 FENG Chun-Fang,WU Gao-Bing,CAO Sha,LIU Zi-Duo and HONG Yu-Zhi 773true 2009/6/30 0:00:00 FENG Yan-Sheng,LIU Mei-Dong,LIU Ying,LIU Jun-Wen,CHEN Guang-Wen,ZHANG Hua-Li and XIAO Xian-Zhong 772true in vitro for research. Methylation status of ANXA1 gene was detected by methylation specific polymerase chain reaction (MSP), while mRNA expression level were also evaluated by reverse transcriptional polymerase chain reaction (RT-PCR). Subsequently, different end concentration (0 μmol/L as control, 0.1 μmol/L, 1 μmol/L, 5 μmol/L,10 μmol/L) of 5-aza-2′-deoxycytidine(5-aza-2dC) were added into culture medium of NPC cell lines for 72 hours′ de-methylation treatment, then methylation status of ANXA1 gene were detected by MSP and mRNA expression were evaluated by RT-PCR. In addition, ANXA1 protein expression was detected by Western-blotting. Without de-methylation treatment, ANXA1 gene was methylated in all the four NPC cell lines but NP69, and the methylation extent is correlated with differentiation state and metastasis potential of the cells. mRNA expression was lower in all of the four NPC cell lines without de-methylation treatment compared with NP69, and the expression level was correlated with gene methylation level. 5-aza-2dC de-methylation treatment reversed ANXA1 methylation status, and increased the expression levels of mRNA and protein in all the four NPC cell lines. In summary, the current research verified down-regulated ANXA1 gene expression in NPC cell lines from both mRNA and protein level, the expression down-regulation was mainly caused by gene methylation, and 5-aza-2dC de-methylation treatment restored the down-regulated expression of ANXA1 in NPC cell lines to the level in the non-neoplastic cell line NP69.]]> 2009/5/13 0:00:00 TAN Shuang-Xiang,HU Rui-Cheng,DAI Ai-Guo,TANG Cen-E,YI Hong,CHENG Ai-Lan,CHEN Zhu-Chu,LI Jian-Ling and XIAO Zhi-Qing 771true in vitro insulin resistant model of liver cells and evaluate the effects and mechanism of FGF-21 on glucose metabolism in the cell model, HepG2 cells were incubated with 10-7 mol/L insulin for 24 h to build insulin-resistant cell model. To evaluate the cells for insulin resistance, the cells were stimulated with fresh insulin for 24 h and the glucose uptake by these cells was carried out. The insulin-resistant cells were treated with different concentrations of FGF-21 for 24 h and insulin-treated cells were used as a control. The glucose uptake by the cells was detected by the method of glucose oxidizes/peroxides (GOD-POD); the synergy between insulin and FGF-21 was evaluated. The mRNA expression of GLUT1 in the insulin-resistant cells was detected by the real-time PCR. Glycogen synthesis of the cells was examined by the anthrone method. The results showed that HepG2 cells treated with 10-7 mol/L insulin for 24 h became resistant to insulin and the insulin resistance status was maintained for 48 h without change of cell morphology. FGF-21 could stimulate glucose consumption of the insulin-resistant model in a dose-dependent manner. The glucose consumption and glycogen synthesis of the insulin-resistant model were significantly improved by FGF-21 treatment. FGF-21 showed strong synergy with insulin in glucose uptake and glycogen synthesis of the model cells. While the cells became resistant to insulin, FGF-21 could increase the mRNA expression of GLUT1. Thus, It is concluded that FGF-21 stimulates glucose uptake in insulin resistant HepG2 cells through GLUT1 expression, stimulates glycogen synthesis and improves the glucose metabolism in the insulin resistant liver cell model.]]> 2009/6/30 0:00:00 LIU Ming-Yao,WANG Wen-Fei,YU Yi-Xue,HOU Yu-Ting,REN Gui-Ping and LI De-Shan 770true 2009/9/14 0:00:00 YANG Cai-Rong,WEI Yan-Chang,ZHANG Yan,ZHENG Ke-Jia,LI Ning and YAN Yun-Qin 769true 2009/5/5 0:00:00 MU Wen-Ying,YU Gang and ZHUANG Feng-Yuan 768true 2010/6/3 0:00:00 ZHANG Jing-Feng,XU Li,WANG Wen-Shan and LI Peng 767true Mash2 is an imprinted gene that plays important role in embryo development and organogenesis. Aberrations were often observed in the lungs of most deceased cloned calves died around birth. In an effort to determine whether the DNA methylation reprogramming of Mash2 is efficient in somatic cloning animals, the DNA methylation status of Mash2 was analyzed in lungs of deceased somatic cloning bovines that died within 48 h of birth using bisulfite sequencing analysis. The results demonstrated that cloned bovines showed significantly lower DNA methylation of Mash2 than controls (P < 0.05), and both showed low methylation (20.04% and 5.55%)，and the percentage of overall mCpG in 9C5 was only 0.4%；the types of methylation patterns were five in 9N3 and one in 9N4，while only one type were found in cloned cow．The abnormal DNA methylation of Mash2 may contribute to the lung development defects and the low efficiency of somatic cloning.]]> 2010/6/10 0:00:00 CHEN Jie,LI Dong-Jie,ZHANG Cui,LI Ning and LI Shi-Jie 766true Geobacillus stearothermophilus CICC 20156 was cloned by bioinformatics technology and then was inserted into the expression vectors pPIC9K and pYG1.2. The recombined plasmids were transformed into yeast Pichia pastoris GS115 and Aspergillus niger M54 respectively. It was revealed by SDS-PAGE and Western blot that the exogenous protein of about 29 ku with a His-tag each were secreted by the transformed hosts into the external mediums. The concentration of recombinant carboxylesterase in medium excreted by Pichia pastoris and Aspergillus niger was 30.7 mg/L and 15.3 mg/L respectively. Bioactivity assay indicated that the carboxylesterase activity was 22 671 U/mg for the enzyme expressed by Pichia pastoris and 21 438 U/mg for that expressed by Aspergillus niger. The research results also showed that both enzymes expressed by the two kinds of hosts possess similar characteristic. The recombinant enzyme exhibited thermostability with optimum temperature at 60℃ and remained 76.7%～67.6% activity even after incubation at 70℃ for 30 min. The enzyme also showed a broader pH tolerance and the optimum pH for the enzyme activity was at 8.0. This is the first report that the thermostable carboxylesterase of Geobacillus stearothermophilus was highly secretively expressed in Aspergillus niger and Pichia pastoris. Though the amount of carboxylesterase excreted by recombined Aspergillus niger was lower than that excreted by recombined Pichia. pastoris, the engineering Aspergillus niger may still show a better perspective for potential applications in biotechnological industries, for no inducer is needed to get crude carboxylesterases in the culture supernatants of it.]]> 2010/7/14 0:00:00 SUN Jin-Xia and LIU Zhong-Bin 765true 2010/6/10 0:00:00 ZHANG Dan-Gui,LI Wei-Zhong,WANG Ge-Fei,ZHANG Heng,ZENG Jun,CHEN You-Ying,ZHANG Chi,ZENG Xiang-Xing and LI Kang-Sheng 764true via the ubiquitin-mediated proteasomal pathway, and PINK1 decreased level of Parkin by promoting its degradation via the ubiquitin-mediated proteasomal pathway. The results confirmed that Parkin directly interacted with PINK1, and they regulated each other via the ubiquitin-mediated proteasomal pathway.]]> 2010/6/29 0:00:00 WANG Xue-Jing,GUO Ji-Feng,JIANG Hong,SHEN Lu and TANG Bei-Sha 763true E. coli or epitope peptides synthesized in vitro as immunogen, protein expression profiling were investigated by Western blotting for root, stem, leaf and panicles at five developmental stages. The results indicated that all chloroplast proteins tested were expressed in leaf, but not detectable in root. The photosynthesis primary reaction protein CAB1 and CAB2, the electron transport protein OEE1, and the ROS scavenging-related proteins 2-CysP and Trx were expressed in stem, but four carbon fixation proteins RCA, GAPDH, FBPA and SBPase, which involved in Calvin cycle, were not detected in stem. In panicle, the chloroplast proteins showed different expression patterns, CAB2 and 2-CysP were expressed at all stages during panicle growth and development, CAB1 and OEE1 were expressed at late stage, and the four proteins involved in Calvin cycle were expressed only in the middle stage. Interestingly, four proteins in Calvin cycle showed the same expression patterns, supporting their cohesive relationship. In addition, the data revealed possible clues of post-translational modification, dimer and different forms of transcripts. Comparison analysis between the profiling of gene transcription and translation revealed parallel phenomena; however, they are quite different at least in some instances. Taking together, this experiment revealed the expression patterns of rice chloroplast proteins in a direct and relative quantitative way, provided helpful information for better understanding their function and also provided a preliminary proof for the concept of a rice antibody-based proteomics strategy.]]> 2010/5/27 0:00:00 BAI Hui,WANG Xian-Yun,CAO Ying-Hao,LI Xiao-Ming,LI Li-Yun,CHEN Hao,LIU Li-Juan,ZHU Jian-Hui and LIU Guo-Zhen 762true 2010/6/3 0:00:00 JIANG Li-Hua,LI Yi-Xue and LIU Qi 761true etc. Moreover, intron sequence and its corresponding coding sequence are existed interaction or co-evolution relations. The relations between intron sequences and its corresponding coding sequences were studied. For the C. elegans ribosomal protein genes, 85 genes were selected from RPG (http://www.cbi.pku.edu.cn/chinese/mirrors.html). The intron sequences were divided into first introns, second introns, other introns, short introns, and long introns and the corresponding coding sequences were divided into exons and all protein coding sequences (CDS), then the matching local alignment between introns and the corresponding coding sequences were done with Smith-Waterman local alignment software. The results show that there are really the interaction regions in introns when it is aligned with coding sequences. When intron sequences are aligned with CDSs, the significant interaction regions for the first intron and the other intron are located in about 15%～55% of intron length and it is located in about 30%～80% of intron length for the second intron. The distribution of interaction regions for short introns is similar to the distribution of the first introns. For long introns, there are two significant interaction regions. The first peak region is located about 15%～30% of intron sequence and the second peak region is located about 54%～78% of intron sequence. When long introns are aligned with exons, there is only one peak region. It is located in about 5%～20% of intron upstream region. When CDS are aligned with every kind of introns, it was found that there are many interaction regions and forbidden regions in CDSs. It was also found that there are two common forbidden regions in the CDSs, they are located at the 10% and 80% of coding sequence. The distribution of interaction regions for the first introns is different from the second introns. When compared the distributions of long introns aligned with CDS and aligned with exons, it can be concluded that the segment of the first peak region are acted on the inner exon segment, the segment of the second peak region are acted mainly on the exon-exon junction regions. Furthermore, there are many peak regions and forbidden regions which are distributed in protein coding sequences. It is speculated that the forbidden regions may be the combined regions of protein complex. In a word, all of the intron sequences besides the 5' end and 3' end correlate closely with their corresponding coding sequences or the two kinds of sequence segments are existed co-evolution relation.]]> 2010/6/17 0:00:00 ZHAO Xiao-Qing,LI Hong and BAO Tonglaga 760true 2010/6/10 0:00:00 ZHENG Jing-Min,ZHU Xiao-Dong,ZHANG Ming-Chao,WANG Jian-Ping,XU Li-Li,CHEN Da-Cheng and LIU Zhi-Hong 759true Nbs1 is a target gene of c-Myc. Chromatin immunoprecipitation assays reveal that c-Myc recruited the histone acetyltransferase p300 complex to the promoter region of Nbs1 gene, resulting in an increased level of acetylated histone H4. Furthermore, it was found that the two proximal E-box elements located in Nbs1 promoter region were essential for the c-Myc binding. Thus, it was concluded that c-Myc attenuates the effect of doxorubicin partially through regulating Nbs1, implicating its roles in repair of DNA breakage induced by doxorubicin.]]> 2010/3/24 0:00:00 SHI Han-Qing,HUANG Hui,ZHANG Xi-Tao,ZHANG Jian-Chao,LUO Xiu-Tan,PENG Xiu-E,HUANG Bai-Qu and LU Jun 758true 2010/4/20 0:00:00 TANG Jian-Hua,CUI Jie-Feng and LIU Yin-Kun 757true in vitro deubiquitinase (DUB) activity. However, the characteristics and functions of DUB activity of NL63 coronavirus PLPs are poorly understood. It was first demonstrated that the core domain of PLP2, but not PLP1, has in vivo DUB activity, and the DUB activity is not dependent on the catalytic residue of D1849. However, the PLP2 DUB activity is significantly reduced when the catalytic sites of C1678 and H1836 were mutated. PLP2 has both in vivo DUB and DeISGylation activity, and PLP2 exhibits DUB activity toward ubiquitinated branched peptides without any specificity for either Lys48 linkages or Lys63 linkages. Furthermore, PLP2, but not PLP1, is the only core domain responsible for the inhibition of both RIG-I and TLR3-dependent induction of IFNα/β expression. Mechanism study demonstrated that PLP2 interacts with the key regulation players of RIG-I and ERIS (also called STING/MITA) of IFN induction pathway, and induces the deubiquitination of RIG-I and ERIS. Overall, these results definitely demonstrated that for the two core domains of PLP responsible for proteolytically processing of N-terminal part of pp1a(1ab) of NL63 coronavirus, PLP2 is the only core domain of PLPs that responsible for DUB activity and IFN antagonists. The studies are currently underway to determine the biological significance of DUB of NL63 coronavirus PLP2 in virus replication and pathogenesis.]]> 2010/5/12 0:00:00 SUN Li,YANG Yu-Dong,LIU Dian-Bo,XING Ya-Ling,CHEN Xiao-Juan and CHEN Zhong-Bin 756true 2010/5/20 0:00:00 HE Qing-Zhi,TUO Qin-Hui,ZENG Huai-Cai,ZHU Bing-Yang,RANG Wei-Qing,TANG Xiao-Qing and LIAO Duan-Fang 755true ABCE1 in 95-D/NCI-H446 lung carcinoma cells, the protein of ABCE1 was knocked-down using an RNAi approach. Effect of siRNA- expressing vector on ABCE1 and E- cadherin protein expression was evaluated in 95-D/NCI-H446 lung carcinoma cells by Western blot and FACS analysis. A transwell collagen invasion assay was used to assess differences in the invasive properties of cells expressing ABCE1. Treatment of the 95-D or NCI-H446 cells with an RNAi to ABCE1 led to near 85% knockdown of ABCE1 expression after 48 h. Interestingly, knockdown of ABCE1 expression was accompanied by a significantly higher level in E-cadherin expression. And the expression of E-cadherin in siRNA-95D/H446 was much higher than that in either of the control cell lines by FACS analysis. Moreover, cells treated with the ABCE1 RNAi displayed poor less invasion than the control cell lines too.There were close relationships between ABCE1 and E- cadherin. It has been suggested that silencing ABCE1 expression increases E-cadherin expression and decreases cell invasion in 95-D / NCI-H446 lung carcinoma cells.]]> 2010/5/12 0:00:00 ZHAO Ai-Jun,ZHENG Mao-Gen,WANG Guo-Chen,HOU Jing-Pu,XU Qi,WANG Jin-Feng,LI Zuo-SHENG and CHEN Zhi-Quan 754true in situ hybridization and acridine orange staining. Whole-mount in situ hybridization with neurog1, elavl3, gfap and krox20 RNA probes showed that the expression of neural precursor cells, neurons, neuroglia cells and rhombencephalon3, 5 was strikingly affected; meanwhile, the expression of MHB (midbrain-hindbrain boundary) markers pax2a and wnt1 was significantly compromised and showed duplication of neural tube in pcdh18b down regulation group. Acridine orange staining pointed out that down regulation of pcdh18b increased cell apoptosis in midbrain, hindbrain and MHB region. These results suggest that pcdh18b plays an important role in the zebrafish neurogenesis.]]> 2010/5/20 0:00:00 GONG Yue-Bo,JIANG Qiu,HU Jing-Ying,WANG Yue-Xiang and SONG Hou-Yan 753true 2010/5/27 0:00:00 CONG Xiao-Jing,TAN　Jian-Jun,LIU Ming,CHEN Wei-Zu and WANG Cun-Xin 752true 2010/5/12 0:00:00 ZHANG Heng,JIA Ya-Li,YUE Wen,CHEN Lin,XIE Xiao-Yan,NAN Xue,HE Li-Juan,PEI Hai-Yun and PEI Xue-Tao 751true Pinctada fucata. It is interesting to investigate the relationships among the oyster engrailed, BMP2 and Smad3 expressions in the oyster. Thereby a partial fragment of an engrailed homolog has been amplified from the genome template of the pearl oyster Pinctada fucata. Alignments with the deduced amino acids sequence show that the DNA-binding homeobox (EH4) domain of the oyster engrailed shares extremely high similarities with other engrailed proteins. Shell notching experiment and semi quantitative polymerase chain reaction results suggest that the change pattern of the oyster engrailed mRNA level is similar to that of the oyster mothers against decapentaplegic 3 homolog (Smad3) in shell repair process. Then the Smad3, the engrailed and the BMP2 mRNA expression levels in primary cultured oyster mantle tissue cells that are treated with dexamethasone (DXM) and hydrogen peroxide (H2O2) respectively have been examined. Real time quantitative polymerase chain reaction results show that the engrailed, the Smad3 and the BMP2 mRNA levels in the oyster mantle cells exhibit significant correlations with each other. All the results provide important clues and basis for further study on developmental and signal transduction mechanisms on oyster biomineralisation.]]> 2010/3/24 0:00:00 ZHOU Yu-Juan,HE Zuo-Xin,LI Chang-Zhong,XIANG Liang,ZHU Fang-Jie,ZHANG Gui-You,XIE Li-Ping and ZHANG Rong-Qing 750true vs 0.277 ± 0.012, P < 0.01) and inhibit its protein expression by 94.9%(0.012 ± 0.005 vs 0.237 ± 0.010, P < 0.01); Sp1 and Ap1 protein were significantly decreased by 14.0% (0.860 ± 0.013 vs 1.000 ±0.015, P < 0.05) and by 22.0% (0.780 ± 0.033 vs 1.000 ± 0.052, P < 0.05), compared with control, respectively. The growth rate of HUVEC treated with the 27-nt microRNA high expression was significantly decreased, particularly the inhibition in the cell lines transfected with double-length and mutant of the 27-nt microRNA plasmid, by which the doubling-time of growth was increased by 49.4% (29.22 ± 0.25 vs 19.55 ± 0.19, P < 0.05), compared with control. The data suggested that intronic 27-nt microRNA significantly inhibit the eNOS expression and the proliferation of HUVEC, by the time the expression of transcription factors Sp-1 and Ap-1 were altered, strongly suggesting that 27-nt microRNA and transcription factors cooperatively regulate the expression of related genes, consequently the proliferation of HUVEC. Data from the present study may serve as one model of the critical mechanisms through which the intronic microRNA and transcription factor synergisticly were involved in the auto-regulation of disease-related gene expression.]]> 2010/3/9 0:00:00 YAN Li-Mei,WU Jian-Yong,YU Xiao-Hua,XU Jing-Ou and Ou He-Sheng 749true 2010/4/28 0:00:00 SONG Ke-Dong,LIU Tian-Qing,ZHAO Guo-Feng,WU Shuang,FANG Mei-Yun,SHI Fang-Xin,ZHU Li-Li,MA Xue-Hu and CUI Zhan-Feng 748true 2010/3/23 0:00:00 YAN Li-Cheng,SU Ji-Guo,CHEN Wei-Zu and WANG Cun-Xin 747true TM) of Uncaria tomentosa has antioxidant activity and beneficial effects on DNA repair and immune function. Batch-2 is a novel aqueous extract from cat's claw, however, the free radical scavenging ability and neuroprotective effect of batch-2 have not been reported. Firstly, the neuroprotective effect of batch-2 on the 6-OHDA-induced SH-SY5Y cell damage was detected. Secondly, the component analysis of batch-2 was tested by infrared spectroscopy, HPLC and spectrophotometry.The results showed that batch-2 has scavenging ability to several kinds of free radicals, especial the hydroxyl radical (25 mg/L of batch-2 can scavenge 60% of the hydroxyl radical). The dose dependently attenuated 6-OHDA-induced cell death, lipid peroxidation, mitochondrial potential loss and increase of intracellular reactive oxygen species (ROS) and nitric oxide (NO) in SH-SY5Y cells were also observed. Meanwhile, batch-2 can suppress the 6-OHDA induced increase of expression of iNOS and activity of NF-κB. The results show that the neuroprotective effect of batch-2 on 6-OHDA-induced damage of SH-SY5Y cells by scavenging ROS and NO, inhibiting activation of iNOS and NF-κB. Component analysis shown that the content of polyphenol and quininic acid from Batch-2 was 6.43% and 0.0958% respectively. These results provided evidence to support that the mechanism of batch-2 for antioxidative activity and neuroprotective effect may be similar with (-)-epigallocatechin-3-gallate(EGCG) partly. It is obvious that batch-2 is a potential neuroprotective antioxidant for the precaution candidate of Parkinson's disease.]]> 2010/6/10 0:00:00 SHI Zhen-Hua,Xi Zhao-Wilson,WANG Hao,GUAN Peng,YU Peng,CHANG Yan-Zhong,DUAN Xiang-Lin and ZHAO Bao-Lu 746true i in N2a/Swe.△9 cells. Oxidative stress induces increased activation of C-JunN-terminal kinase (JNK). It was demonstrated that anthocyanin can decrease the activation of JNK in N2a/Swe.△9 cells, suggesting that anthocyanin exerts its protective effect by inhibiting the activation of JNK. Therefore, anthocyanin could act as an oxidative stress suppressor in protecting the N2a/Swe.△9 cells against Aβ induced cell injury and it is a promising candidate for AD treatment in the future.]]> 2010/3/24 0:00:00 LIU Ling-ling,SHENG Bai-yang,YAN Yu-fang,GONG Kai,MA Tuo,ZHAO Nan-ming,ZHANG Xiu-fang and GONG Yan-dao 745true 2010/5/20 0:00:00 SHI Liang,WU Rui-Jie,XU Cui-Ping,ZHANG Shou-Wen,ZHU Hong-Wei,LIU Zu-Xiang and LI Yong-Jie 744true ras gene mutation is generally regarded as a significant indicator for colorectal cancer screening and early diagnosis and in favor of filtering out colorectal cancer patients benefiting from epidermal growth factor receptor (EGFR)-targeted therapy. The chip-based temperature gradient capillary electrophoresis (TGCE) was successfully used to detect K-ras gene mutation in 98 cases of paraffin-embedded colorectal cancer tissues with high sensitivity. Mutation-positive detection rate from chip-based TGCE (47.96%) was significantly higher than that from the PCR product direct sequencing (23.47%). Clone sequencing showed that this detection system could detect at least 2.08% of the mutant K-ras gene. Analysis of the relationship between K-ras gene mutation and clinical pathological parameters revealed that K-ras gene mutation rate in rectum cancer was significantly higher than that of colon cancer (P < 0.05), but no remarkable correlation between K-ras gene mutation rate with age, gender, histological type and tumor stage was observed. Therefore chip-based TGCE is very sensitive and rapid method for mutation detection, and it can be applied for large-scale screening, early diagnosis and guiding clinical treatment.]]> 2010/3/23 0:00:00 ZHANG Hui-Dan,WANG Xiao-Nan,ZHOU Zhe,MA Qian and FANG Jin 743true Hipposideros armiger) and effect of the recovery cycle on the following pulse repetition rate were studied using mimic CF-FM sound stimuli emitted by free flying bat. Recovery cycle of 93 IC neurons were obtained from IC of five bats with normal hearing. These neurons were classified into three types, i.e. long recovery (LR, 47.4%), moderate recovery (MR, 35.1%), and short recovery (SR, 17.5%), according to their inter pulse interval (IPI) (ms) of 50% recovery under two CF-FM sound stimulation condition. Each type of the neurons could also be categorized into different sub-types according to changes induced by IPI increasing such as single-IPI response area neurons, multi-IPI response area neurons, and monotonic-IPI response neurons. Mean IPIs of 50% recovery of LR, MR, and SR neurons were (64.0 ± 24.8), (19.6 ± 5.8), and (7.1 ± 2.4) ms, respectively (P < 0.001). The calculated theoretically following pulse repetition rate (pulse per second, Hz) of LR, MR, and SR neurons by mean IPI of 50% recovery for each type were (18.2 ± 7.0), (55.4 ± 15.7), and (171.3 ± 102.9) Hz, respectively (P < 0.001). These three types of IC neurons were well corresponding to their three hunting phases, search, approach, and catch phases. The sub-types of single-IPI response area neurons and multi-IPI response area neurons had hunting phase selectivity, and sub-type of monotonic-IPI response neurons had well sensitivity to IPI change, but their hunting phase selectivity was of a sort. These results demonstrated that recovery cycle of IC neurons determined the ability to follow pulse repetition rate and matched this bat's echolocation behavior.]]> 2010/3/23 0:00:00 TANG Jia,FU Zi-Ying,JEN Philip H.-S and CHEN Qi-Cai 742true INK4a plays a key role in control of cell cycle progression by negatively regulating the CDK4/6 activity. It was shown that histone acetyltransferase p300 had a positive effect on the activation of p16INK4a promoter, whereas, histone deacetylases HDAC3/4 counteracted the p300-mediated activation of p16INK4a promoter, and decreased the p16INK4a mRNA and protein levels. Chromatin immunoprecipitation (ChIP) tests revealed that the transfection of p300 reversed the hypoacetylation status of histones at the p16INK4a promoter mediated by HDAC3/4. Moreover, the immunofluorescence study showed that the nucleo-cytoplasmic shuttling of HDAC4 may play an important role. Furthermore, Western blot and ChIP assays demonstrated that the HDAC inhibitor sodium butyrate (NaBu) enhanced p16INK4a expression through inducing histone hyperacetylation. Based on these data, a hypothetical model was proposed for the involvement of reversible histone acetylation in transcriptional regulation of the p16INK4a gene.]]> 2010/2/23 0:00:00 WANG Xiu-Li,FENG Yun-Peng,ZHAO Jing,ZHANG Guo-Ping,PAN Hong,HUANG Bai-Qu and LU Jun 741true 2010/3/23 0:00:00 YIN Hui-Qun,CAO Hong-Guo,SUN Xue-Ping,XUE Yi-Jie,ZHANG Wei-Qin,HUANG Wei-Ling,TAO Yong,LIU Ya,LI Yun-Sheng,ZHANG Yun-Hai and ZHANG Xiao-Rong 740true 2010/3/23 0:00:00 ZHU Jing,GUO Xing-yan,MA Yuan and REN Fei 739true EC50=30.9 μg/L. H22 itself was not remarkably associated with the normal, rapid mobilization of calcium from intracellular stores. Instead, it blocked calcium mobilization induced by endogenous chemokines. It proves that using chemokine sequence of human source to build a peptide library and screen effective sequence is available. The conservative sequence could simulate the binding mimotope of Human MIP-1β interacting with HCMV US28 N-terminus to bind with synthesized peptide H22. The study has demonstrated that peptide H22 cannot stop the biological function of hMIP-1β and block the signal transduction from hMIP-1β by the way of calcium pathway, but itself did not affect cells activity.]]> 2010/3/11 0:00:00 LI Lu,HE Tao,MO Xue-Mei,LI Xiu-Ying,ZHANG Guang and SUN Han-Xiao 738true 2010/3/24 0:00:00 ZHANG Jing,LI Wei,ZHANG Kang-Jian,LIU Xi-Jun,KONG Yan-Ping,NIU Na,JIANG Hai and ZHOU Xiu-Mei 737true IC50 was decreased significantly in K562/Dox cells, and the reversal index were 1.76 and 1.73, respectively. Furthermore, animal studies found that, compared with control group, ADT was significantly higher (P < 0.05) and ADD shortened in PHT-CBZ resistant rats after XIAP antisense oligonucleotides was applied. It is obvious that the expression of XIAP was significantly increased in drug resistant K562/Dox cells. Down-regulation of XIAP could reverse the drug-resistance of K562/Dox cells, and improve the electrobiological activity in PHT-CBZ resistant rats. These findings indicate that XIAP is involved in multidrug resistance in medically intractable epilepsy.]]> 2010/1/26 0:00:00 CHEN Ying-Hui,WANG Nai-Dong,ZU Heng-Bing and HONG Zhen 736true 2010/2/23 0:00:00 XU Can-Xin,WANG Chun,ZHU Bing-Yang,GAO Zhi-Ping,LUO Di-Xian and LIAO Duan-Fang 735true via various in vivo assay systems. The inhibition effect of TFP on sarcoma 180 tumor (S180)-bearing mice was observed at dosages of 75, 150 and 300 mg/kg by systematically to measure the S180 tumor inhibition rate, mice survival rate and cellular immunity. The result showed that continuous administration of TFP for 10 day continuously was found to inhibit the growth of transplanted S180, prolong the mice survival days, promote the plasma interleukin-2, interferon-γ levels and improve the T-lymphocyte subsets CD4+ and CD4+/CD8+ percentages after treatment with TFP. In addition, TFP was found to increase the delayed-type hypersensitivity response and macrophage phagocytosis significantly. These results strongly suggest that TFP enhances the host defense response to tumor due in part to the immunomodulatory activity.]]> 2010/1/26 0:00:00 HAN Quan,LING Ze-Jie,HE Pu-Ming and XIONG Chang-Yun 734true 45 is the active site of rBTI. These inhibitors showed remarkable stability to heat and pH. The possible effects of aBTI and fBTI on the proliferation of human HL-60 and EC9706 cell lines were investigated by MTT assays. It indicated that aBTI and fBTI could specifically inhibit the growth of HL-60 and EC9706 cells in a dose- and time-dependent manner. The active site of rBTI was determined and two new inhibitors were obtained in this research. Knowledge about the active site is useful for further clarifying the physiological mechanism of rBTI and other protease inhibitors. New inhibitory activities of aBTI and fBTI are potentially useful in the fields of health and medicine.]]> 2010/1/21 0:00:00 TIAN Xin,LI Chen,LI Yu-Ying and WANG Zhuan-Hua 733true BADH2 and dicot Arabidopsis GR7 were detected by Agrobacterium-mediated tobacco transient expression system. The results indicated that the fundamental splicing regulatory elements are conserved in plants. The tobacco transient expression system could be used as an important tool for fast and sensitive detection of gene splicing regulation in higher plants.]]> 2010/3/9 0:00:00 GAO Shao-Pei,NIU Xiang-Li,LUO Di,CHANG Li-Juan,YU Xu,LI Yu-Xiang and LIU Yong-Sheng 732true OsWRKY89, a rice WRKY transcription factor gene, is induced by UV-B radiation. To identify the cis-elements in the OsWRKY89 promoter related to UV-B response, the promoter and its deleted fragments were each fused with gus reporter gene and transformed into rice calli. The transgenic plants regenerated were used to analyze their responses to UV-B irradiation. Finally, an UV-B response cis-regulatory element was narrowed to the region of 25 bp between -1 188 and -1 213 upstream the translation start site of OsWRKY89 gene, including nucleotide sequence of AAGATCTACCATTGCTCTATAGCTT. Through analysis of the promoter of OsWRKY89 and 28 genes up-regulated by UV-B treatment, two relatively conserved UV-B response elements in the 1 500 bp protmoter region studied were located. Coincidental, the 25 bp of OsWRKY89 promoter contains one of the conserved elements. Furthermore, several conserved light-response elements including ACEOsWRKY89, MREOsWRKY89 and UVBoxAtANAC13 were also found to adjoine to the 25 bp UV-B response region in the OsWRKY89 promoter, suggesting they together play roles in regulating the light responses of the gene.]]> 2010/4/20 0:00:00 ZHAO Chang-Jiang,HAO Zhong-Na,WANG Hai-Hua,CHEN Xu-Jun and GUO Ze-Jian 731true in vivo. IL-10, a cytokine synthesis inhibitory factor, plays an important role in anti-inflammatory response. Here BMSCs were infected with recombinant adenovirus encoding murine IL-10 (mIL-10) and a GFP marker to obtain GFP positive BMSC-mIL-10 cells. The BMSC-mIL-10 cells were injected into mice with cecal ligation and puncture (CLP)-induced sepsis, and the enhanced expression of IL-10 in blood was confirmed by ELISA. Indeed, in CLP mice mode, the systematic delivery of IL-10 via BMSCs effectively suppressed the production of proinflammatory cytokines, including TNF-α, IL-6, IL-1α and IL-1β, compared to BMSCs alone. The therapeutic benefits were further demonstrated by an increased prevention from body loss, increased survival rate, and the suppression of inflammatory response in lung and kidney. Furthermore, these effects may be mediated by inhibiting the activation of NF-κB in macrophages and neutrophils. Collectively, these results suggest that systemic delivery of IL-10 by BMSCs may serve as a potential treatment in sepsis therapy.]]> 2010/5/7 0:00:00 BI Liang-Kuan,TANG Bing,ZHU Bin,XIE Chun-Ling,LI Shuang,LIN Tian-Xin,HUANG Jian,ZHANG Wei and ZHU Jia-Yuan 730true in vitro and in vivo by decreasing the expression of Wnt/β-catenin signal pathway molecules.]]> 2010/2/4 0:00:00 SUN Xiao-Jie,ZHOU Qi-Bing,ZHAO Mei,LI Shen,GUO Hong-Yan,WU Qi and HUANG Chang-Zhi 729true 2010/2/4 0:00:00 YE Ling,ZHANG Gui-Ying,LIU Ting,CHEN Xuan-Min,YI Hong,XIAO Zhi-Qiang,LENG Ai-Min and PENG Jie 728true 2010/3/9 0:00:00 WU Feng-Ying,FENG Qiong,CHENG Min,YAN Jie,XU Yu-Xia and ZHU Cui-Qing 727true 2010/2/4 0:00:00 CHEN Li-Na,WANG Qian,SHANG Yu-Kui,ZHANG Liang-Cai,SUN Zhao,HE Wei-Ming,ZHAO Yan,LI Wan,WANG Hong,HE Yue-Han and LI-Xia 726true 2010/1/21 0:00:00 ZHENG Hua-Yang,TANG Bi-Kui,WANG Pei-Wei,GAO Fang-Jian,HU Xiao-Bo,GONG Li and ZHU Nai-Shuo 725true 2010/1/21 0:00:00 ZHANG Xuan,YE Li-Hong and ZHANG Xiao-Dong 724true 2010/1/19 0:00:00 CAO Dong-Li,YIN Kai,MO Zong-Chen,HAO Xin-Rui,HU Yan-Wei,LI Xiao-Xu,TANG Ya-Ling and TANG Chao-Ke 723true Anabaena sp. PCC 7120 bound PCB with reconstitution in E. coli, the λmax of absorption and fluorescence spectra were 605 nm and 633 nm before purification, while the λmax of absorption and fluorescence spectra were 650 nm and 665 nm after purification. To study the phenomenon above, eight mutants were constructed. It is indicated with reconstitution in E. coli that: the mutant ApcD (Y88I) had one more absorbance and fluorescence peaks after purification , the λmax of absorption and fluorescence spectra were 668 nm and 690 nm. The spectra of ApcD (W59Q), ApcD (Y73A), ApcD (W87E) had no change during the purification. The absorption spectra of ApcD (M126S), ApcD (Y116S), ApcD (M160T) had no change during the purification, but the fluorescence spectra had red shifts by 5 nm, 7 nm and 10 nm after purification, respectively. The λmax of absorption and fluorescence spectra of ApcD (M115I) had changed from 605 nm and 633 nm to 638 nm and 655 nm. Under acidic urea conditions, those chromoproteins had maximal absorption at 662 nm, indicating that they had PCB chromophore. Under the study of CD spectra of PCB-ApcD, PCB-ApcD (Y116S) and PCB-ApcD (M160T), the two mutants influenced the conformation of chromophore which bound with apoprotein, but not affected the secondary structure of the reconstitution proteins.]]> 2010/1/26 0:00:00 WANG Xing,ZHANG Qian,YANG Bei,ZHAO Kai-Hong and ZHOU Ming 722true Bacillus sp. I121 was used to study the salt-stress responding proteins. Differential proteomic analysis of the purified plasma membrane fractions were performed by two dimensional Blue-Native/SDS-PAGE. Eight proteins corresponding to NaCl were identified by MALDI-TOF/TOF. Up-regulated proteins under salt stress include an ABC transporter permease, glycerol-3-phosphate permease, pyrimidine nucleotide transporter, formate dehydrogenase, and down-regulated proteins include succinate dehydrogenase iron-sulfur subunit, flavoprotein subunit, cytochrome b-556 subunit, and a hypothetical membrane protein similar to charperone DnaJ. Some of these changes were further verified by enzyme activity assay. Most of these proteins are highly hydrophobic trans-membrane protein, and responsible for material transport across membrane and energy metabolism. These results indicated that in halophilic Bacillus sp. I121, higher salt stress activated material transport across plasma membrane, and compatible solutes, such as proline and ectoine, could be synthesized through suppressed TCA cycle. The research further demonstrated that besides the protein complexes in mitochondria and chloroplast, BN/SDS-PAGE is also a powerful tool for the protein complexes in plasma membrane.]]> 2010/3/22 0:00:00 LIU Xiao-Yu,LI Liang,ZHAO Xin,CHENG Yan-Wei and ZHANG Wei 721true Spirulina platensis by ultra-filtration, gel filtration chromatography and reverse-phase high-performance liquid chromatography. The purified peptide was identified by matrix assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) and amino acid sequencing. Furthermore, the inhibition pattern of the peptide was also investigated and the stability was evaluated under simulated gastrointestinal condition. The results demonstrated that the digests with molecular mass ranging from 0 to 3 000 ku had the most potent ACE inhibitory activity with an IC50 value of (1.03 ± 0.04) g/L. An ACE inhibitory peptide with an IC50 value of (0.009 4 ± 0.000 2) g/L which was equivalent to (27.36 ± 0.14) μmol/L was obtained from that fraction of digests, and was identified as Val-Glu-Pro. The Lineweaver-Burk plot and the Dixon plot indicated the ACE inhibitory peptide was a non-competitive inhibitor with a Ki value of (23.59 ± 0.54) μmol/L. In vitro stability assay showed that the peptide could keep its inhibitory activity well after incubation with gastrointestinal proteases including pepsin, chymotrypsin, and trypsin, suggesting the ACE inhibitory peptide from Spirulina platensis be of great prospects as an ingredient of functional foods or pharmaceuticals in prevention and treatment of hypertension.]]> 2009/12/15 0:00:00 LU Jun,REN Di-Feng,WANG Jian-Zhong and TANOKURA Masaru 720true L by TNF-α induced. Further results show that BimL displaces Bcl-xL in the mitochondria and promotes Bax translocation during TNF-α-induced apoptosis. Furthermore, SP600125 (specific inhibitor of JNK) can inhibit the Bax translocation to mitochondria. Finally, Bax is found to translocate to mitochondria in Naive PC12 cells with co-expressing of GFP-BimL and YFP-Bax. The research demonstrates the important role of BimL, and reveals that BimL activate Bax indirectly during TNF-α-induced apoptosis.]]> 2010/1/27 0:00:00 ZHANG Lan and XING Da 719true 2009/11/30 0:00:00 YANG Chao,JI Lei,SHI Shuang-Shuang,YUE Wen,HE Li-Juan,NAN Xue and PEI Xue-Tao 718true 2009/12/23 0:00:00 ZHANG Ying-Ying,JIN Xin,GONG Hai-Qing and LIANG Pei-Ji 717true 2009/12/23 0:00:00 LIU Ying,JIANG Hong,BAI Jing and GONG Cheng-Xin 716true 2010/1/27 0:00:00 LIU Qi-Jun,WANG Zheng-Hua,LIU Wan-Lin,LI Dong,HE Fu-Chu and ZHU Yun-Ping 715true n=3, P < 0.05); b. Expression of rVBMDMP gene regulated by hTERT promotor was abled to inhibit the growth of lung cancer A549 cells and transplanted tumor in nude mice (n=6, P < 0.05); c. The expression of rVBMDMP gene promoted the apoptosis of lung cancer A549 cells and the expression of Caspase-3 protein. These results suggested that expression of rVBMDMP gene regulated by hTERT promotor inhibited the growth of lung cancer A549 cells because it enhanced expression level of Caspase-3 protein to increase cell apoptosis.]]> 2010/2/4 0:00:00 ZHANG Zhi-Wei,HE Xiu-Sheng,LUO Zhao-Yang,LI Yan-Lan and CAO Jian-Guo 714true 2+, the specific nicking activity of T7EⅠ-SAD to pUC(AT) and random nicking activity to pUC19 are close to that of wide-type enzyme. In buffer with Mn2+, the random nicking activity of T7EⅠ-SAD is higher than wide-type T7EⅠ significantly. Results of stepwise denaturing and elution experiment showed that the subunits of T7EⅠ-SAD can disassociate in 5～6 mol/L urea or 1.75～2.0 mol/L guanidine hydrochloride. Gel-shift assay showed T7EⅠ-SAD can binding to four-way junction substrate Junction3 specifically. This study provides an efficient strategy to prepare cytotoxic protein, it provides a sample intuitive approach to probe the stability of protein dimerization too.]]> 2010/1/21 0:00:00 FAN San-Hong,SHAN Li-Wei,WANG Bao-Li,GUAN Chu-Di and GUO Ai-Guang 713true 3O4 particle was synthesized by coprecipitation, magnetic polymeric microspheres- magnetic chitosan microspheres, containing a magnetic core and a polymer shell, were prepared by the suspension cross-linking technique. Novel affinity magnetic microspheres were obtained after immobilizing heparin onto the nanoparticles. The size, morphology, structure and magnetic response properties were studied by means of SEM, FT-IR and XRD and so on. The novel affinity magnetic microspheres′ properties for thrombin were examined by its adsorption and purification, and compared with the traditional means of column chromatography—DEAE-Sepharose Fast Flow. The results showed the affinity magnetic microspheres had a spherical appearance, narrow distribution of diameter, the particle diameter is about 50 nm, and one-step purification of thrombin using it the specific activity was 1 879.7 U/mg, recovery rate was 85% and purification fold was 11.057. Whereas, purification of thrombin by column chromatography, the product exhibits an activity of 909.84 U/mg, recovery rate of 72% and purification fold of 5.33. In conclusion, chitosan nanometer affinity magnetic microspheres was prepared and magnetic separation techniques was applied for purification of thrombin, the results is better than that from traditional methods and these results have some references values in the production and subsequent study on the research and development.]]> 2009/11/30 0:00:00 LI Mei-Ji,LI Zhao-Hua,HU Jian-Cheng and DONG Xian-Zhi 712true Escherichia coli BL21 (DE3). Furthermore, the expressed HSBP1 protein was purified and crystallized. The distinct crystal form was obtained by the hanging-drop vapor-diffusion process to carry on screening and the optimization to the crystallization condition. The crystals belong to R3 space group with the cell parameter a = b =35.2Å, c= 233.3Å.]]> 2010/1/21 0:00:00 ZHANG Hong-Yan,LIU Xue-Qi,SUN Yu-Na,HAO Ning,LI Xue-Mei and LOU Zhi-Yong 711true LDHC gene in male infertility, 100 male patients with unexplained infertility were screened by activity staining of LDH-C4 on sperm smears and those with lowered or no LDH-C4 activity in their sperms were subject to DNA analysis. The encoding exons were PCR amplified and the PCR products were screened by DHPLC for mutations. The PCR products with abnormal elution peaks were subject to DNA sequencing. In 14 infertile male patients with lowered LDH-C4 activity in sperms, it was identified a heterozygous T > A mutation in base 115 of exon 5 of LDHC gene(GenBank accession number: GU479375), changing codon 178 (TTG, encoding leucine) to a stop codon (TAG). This nonsense mutation (L178X) was predicted to result in a severely truncated protein. The mutation and the heterozygosity were further confirmed by T-cloning. To the best of the knowledge, this is the first mutation found in human LDHC gene, substantiating that mutations in LDHC gene might cause male infertility.]]> 2010/2/8 0:00:00 LI Bo,ZHANG Duo,ZENG Jian,FU Xian-Guo,KE Long-Feng,YAN Shui-Di,YAN Ai-Zhen and LAN Feng-Hua 710true FDDV12 transmembrane protein (gp45) had a high-frequent translational terminating mutation at the site of 261W, resulting in a truncation of 154 amino acid residues at the C-terminus. To explore the biological meaning of the gp45 truncation, a gp45-truncated molecular clone was constructed by using an infectious clone of EIAVFDDV12 as the backbone. Replications features of the gp45-truncated EIAV and its prototype virus were analyzed and compared in cultivated monocyte-derived macrophages (MDM) of equine and donkey and fetal donkey dermal cells (FDD). Results showed that the replication capacity of the gp45-truncated EIAV in equine and donkey MDMs was significantly decreased compared to the untruncated gp45 EIAV(P < 0.01), especially in the horse macrophages. In contrast, the truncated EIAV replicated significantly faster than the untruncated EIAV in FDD cells (P < 0.01). In addition, the reduced replication of the gp45-truncated EIAV in equine MDM led to a significant decline of cytotoxicity of the host cells when compared with the gp45 untruncated EIAV (P < 0.05). These results suggest that the truncation of the 154-residue C-terminus of the EIAV gp45 glycoprotein was an adaptation to the attenuation of the EIAV vaccine strain in FDD cells. This truncated mutation reduces the replication of the vaccine strain in macrophages, the primary in vivo target cell of EIAV, which leads to a further attenuation of the vaccine.]]> 2009/12/18 0:00:00 JIANG Cheng-Gang,MA Jian,GAO Xu,LIN Yue-Zhi,ZHAO Li-Ping,HUA Yue-Ping,LIU Di and ZHOU Jian-Hua 709true 2009/12/29 0:00:00 HE Qun,LI Chun-Hui,PAN Zhong-Cheng,WANG Tian-Jiao,ZHANG Yu-Kui,ZHONG Lian-Sheng,WANG Shao-Cheng and ZHAO Yu-Jie 708true pSilencer2.0-U6 to construction of eukaryotic expression vector of siRNA specific for targeting PCNA gene, the negative plasmid pShPNA was also constructed. The plasmids were identified by sequence analysis. The plasmid was transiently transfected into hepatocelluar carcinoma HepG2 cells for 48 h, proliferation and the clone formation of HepG2 cells were detected by CCK-8 and clone formation assay respectively. The percentage of hypodiploid cells and early apoptotic cells was detected by flow cytometry. The morphology was examined by fluorescence microscope after Hoechst 33258 staining. Compared with control group and negative control group, PCNA mRNA level was reduced by pShPCNA detected by RT-PCR. The proliferation and clone formation of HepG2 cells treated with pShPCNA for 48 h were significantly inhibited (P < 0.01). The migration was also attenuated when PCNA was knocked down by shRNA, and flow cytometry analysis showed an increase of the percentage of G0/G1 phase cells, along with a decrease of cell population in the S phase. Percentages of hypodiploid cells and early apoptosis rates were significantly higher in treatment groups than those in control and negative control group (P < 0.01). Mitochondrial membrane potential was reduced in pShPCNA group detected by JC-1 fluorescent staining. Apoptotic morphology such as cell shrinkage, nuclear condensation, nuclear fragmentation, chromatin condensation and apoptotic bodies were also observed by staining with Hoechst33258 under fluorescence microscope. The study indicated the eukaryotic expression vector, PCNA shRNA has been successfully constructed, and effectively inhibits proliferation and induces apoptosis and arrested HepG2 cells in G0/G1 phage.]]> 2009/11/30 0:00:00 ZHANG Xiao-Ying,WU Feng-Yun,HE Jin-Hua,LIAO Xiao-Li,WANG Wei and JIANG Jian-Wei 707true in vitro, a TRAIL gene expressing eukaryotic vector and a short interfering RNA(siRNA) expressing vector were constructed. 72h after transfecting with each vector, semi-quantitative reverse transcription polymerase chain reaction (semi-qRT-PCR) and Western blotting were applied to detect the expression levels of TRAIL mRNA and protein, respectively in DSC. Besides, MTT assay was performed to investigate the cell growth activity and proliferation capacity, and flow cytometry was used to examine cell cycle distribution and apoptosis rate of DSC. Restriction endonuclease digestion and sequencing of nucleotide acid confirmed the correct construction of both over-expression and interfering vectors. The combined results from TRAIL overexpression and interfering in DSC demonstrated that TRAIL is capable of markedly inhibiting DSC proliferation by blocking most of cells at G0/G1 phase, meanwhile the apoptosis rate of DSC was notably increased.]]> 2009/12/29 0:00:00 ZHANG Jin-Ping,LUO Wen-Ping,ZHANG Qian,PENG Hong-Ying,TAN Dong-Mei,WANG Ying-Xiong and TAN Yi 706true 2009/12/24 0:00:00 LIU Xiao-Mu,SONG En-Liang,LIU Gui-Fen,WU Gong-Ying,TAN Xiu-Wen,LIU Zhen-Shan and WAN Fa-Chun 705true Igf-1 gene, which is mainly expressed in stretched osteoblasts. The main purpose of this study is to examine the effects of MGF and E peptide (the C-terminal 24 amino acids peptide in the E domain of MGF, MGF-Ct24E) on differentiation of osteoblast MC3T3-E1 cells. The results indicate that the MGF and MGF-Ct24E activated the extracellular signal-regulated kinase 1/2(Erk1/2) and affected the expression of osteoblast-associated genes, including the reduced expression of alkaline phosphatase (ALP) and type 1 collagen (Col1), the increased product of osteopontin (OPN) and the decreased nuclear levels of activated Cbfa-1. These effects could be abolished by the blockade of Erk1/2 activation by inhibitor PD98059. In addition, the MGF could preferably activate the PI3K-Akt pathway, which is essential for the differentiation of osteoblasts, so the osteoblast has higher expression of ALP, type 1 Col1, and the formation of bone mineralization nodule under the treatment of MGF than that under the treatment of MGF-Ct24E. These advantages could be inhibited after blockading of Akt by inhibitor LY294002. It was concluded that the MGF-Ct24E could inhibit osteoblasts differentiation through the activated Erk1/2 cascade, while the MGF can induce differentiation through the activated PI3K-Akt. The MGF consists of MGF-Ct24E and IGF-1, which could activate Erk1/2 and PI3K-Akt respectively. Therefore, the MGF possess double effects on differentiation of osteoblasts, which presented as that inhibitory effect at the early differentiation, and promoting effect at the later differentiation.]]> 2010/1/7 0:00:00 ZHANG Bing-Bing,WANG Yuan-Liang,YANG Li,PAN Chang-Jiang,FU Ya,DENG Mo-Yuan and LI Yu-Xiao 704true 2+ concentration. The results show that the reactive oxygen species (ROS) and cytosolic Ca2+ concentration at 0.1mT, 0.5mT and 1.0 mT EMF have be all signification increaser (P < 0.01) than control (sham exposure), ROS at 0.1mT and 0.5mT and Ca2+ at 0.1mT have be no difference (P > 0.05) than Trolox + EMF, ROS at 1.0 mT and Ca2+ at 0.5mT and 1.0 mT have be signification increaser (P < 0.01) than Tolox + EMF. It is known that EMF may enhance ROS formation in neurons, and there is positive correlation between ROS level and cytosolic Ca2+ concentration.]]> 2010/1/7 0:00:00 SU Hai-Feng,BAO Jia-Li and LI Peng 703true 2009/12/18 0:00:00 CAO Zan-Xia and WANG Ji-Hua 702true 2009/11/25 0:00:00 JIANG Qi-Feng,CAI Shao-Xi and YAN Xiao-Qing 701true DHFR) in the development of pharyngeal arches were explored. In DHFR knock-down embryos, the malformations of pharyngeal arches and palates were showed by paraffin section and alcian blue staining. DHFR over-expressing can rescue these malformations. TBX1 and HAND2 are two key transcription factors in the development of pharyngeal arches. Whole-mount in situ hybridization and Real-time PCR were performed to detect the expression levels of TBX1 and HAND2. The expressions of TBX1 and HAND2 were reduced in DHFR knock-down group. DHFR over-expression can increase expressions of TBX1 and HAND2. These results demonstrated that DHFR was essential for the development of pharyngeal arches and DHFR regulated the development of pharyngeal arches by effecting the expressions of TBX1 and HAND2.]]> 2009/12/28 0:00:00 SUN Shu-Na,GUI Yong－Hao,JIANG Qiu,QIAN Lin-Xi and SONG Hou-Yan 700true M146V/+ mice showed more reduced complexⅣ activity and ATP content than Tg2576 mice. These data indicated that the dose-dependent effect of Aβ on mitochondrial dysfunction.]]> 2009/11/30 0:00:00 LIU Ling-Ling,SHENG Bai-Yang,GONG Kai,ZHAO Nan-Ming,ZHANG Xiu-Fang,TANG Pei-Fu and GONG Yan-Dao 699true in vitro and interacted with 9G8. Overexpression of GSK-3β inhibited 9G8-mediated tau exon 10 inclusion.]]> 2009/12/15 0:00:00 DING Shao-Hong,YIN Xiao-Min,SHI Jian-Hua,QIAN Wei and LIU Fei 698true via RT-PCR assay. Dox was used to induce the expression of NPCEDRG and a cell clone sensitive to Dox was selected. The best-induced concentration was determined with different concentration of Dox induction. Cell morphology observation, growth curves, clone formation rate and cell cycle distribution were detected after NPCEDRG restoration expression with Dox induction. After the restoration of NPCEDRG expression, the degree of CNE2 differentiation was higher than ever, the growth capacity and clone formation potential of CNE2 cells in soft agar were significantly suppressed, and the cell percentage in G0/G1 phase increased, while percentage of cells entering the S and G2 phase decreased. This data indicates that an abnormality of NPCEDRG expression is associated with nasopharyngeal carcinogenesis and that it may play an important role in inducing cell differentiation, controlling cell growth and regulating the cell cycle.]]> 2010/1/7 0:00:00 YANG Shuai,HU Hua,DENG Min,DONG Juan-Hui,WANG Yan,LUO Qiao,HE Xiu-Sheng and CHEN Zhu-Chu 697true P < 0.05), while the expression of p-p38MAPK mRNA decreased after anti-Lewis y antibody treatment (P < 0.05). FCM showed that the apoptosis rate increased in SB203580 treated RMG-I-H cells(P < 0.05). The mRNA level of p38MAPK and caspase-3 increased by treatment with Carboplatin. The mRNA level of caspase-3 also elevated by treatment with SB203580. In conclusion, high expression of Lewis y inhibits apoptosis in ovarian cancer cells, probably due to involvement of Lewis y in regulating p38MAPK signaling pathway, thereby causing drug resistance.]]> 2009/12/15 0:00:00 CONG Jian-Ping,LIN Bei,LIU Juan-Juan,LIU Qing,LI Fei-Fei,LIU Shui-Ce,GAO Song and ZHANG Shu-Lan 696true P < 0.05). Then it was investigated whether Chk1 and Chk2 proteins could influence cell cycle regulation by knocking down their expressions in BGC823 cells. Further investigation revealed that inhibition of the Chk1 expression in Chk1 siRNA transfected group cut down the proportion of the cells in G2/M phase from 58.1% to 10.4% in BGC823 cells after 24 h induced by DADS (P < 0.05).While inhibition of the Chk2 expression in Chk2 siRNA transfected group had little effect on G2/M arrest after treatment with DADS(P > 0.05). Western blot showed that although DADS decreased expression of CDC25C and cyclinB1 in untransfected cells, inhibition of expression of CDC25C and cyclinB1 treated by DADS was blocked by Chk1 gene silence (P < 0.05). On the contrary, Chk2 gene silence can not do so. These results suggest Chk1 gene silence could abrogate G2/M arrest induced by DADS in BGC823 cell line, and Chk1/CDC25C/cyclinB1 pathway was involved at the G2/M arrest induced by DADS.]]> 2009/12/11 0:00:00 LING Hui,LU Li-Feng,WEN Ling,HE Jie,TAN Hui,XIA Hong and SU Qi 695true 2009/12/11 0:00:00 ZHOU Shou-Hong,YANG Xu-Hong,WU Shu-Jin,CHEN Gen-Grong and LIU Li-Ying 694true Pichia pastoris cell surface display system, two new systems based on two different anchor proteins derived from Saccharomyces cerevisiae Flo1 protein (Flo1p) were constructed respectively. The N-terminal anchor system could make the foreign lipase displayed on the P. pastoris cell surface with its C terminus free by fusion with an anchor protein containing N-terminal flocculation functional domain of Flo1p (874 residues, FS) , which was able to adhere to the cell surface via noncovalent interaction with the mannan chain of the cell wall. Conversely, the foreign lipase can kept its N terminus free in another C-terminal anchor system, which utilizes a GPI-attachment signal domain located at C-terminal region of Flo1p (1 101 residues, FL) as anchor protein. Using these systems above, recombinant R. miehei lipase with a pro region (ProRML) , which had its active site near the C-terminus, was displayed on the P. pastoris cell surface, and two surface-displayed RML, named as FSR and FLR, were obtained. Cell-surface display of the RML via Fs or FL anchor system was confirmed by flow cytometer and laser scanning confocal microscope. A strong fluorescence was clearly observed in recombinant yeast cells harboring pKFSR( pKFS-RML), but no fluorescence was detected in the yeast cells harboring pKFLR(pKFL-RML) . The hydrolytic activity of FSR reached 105. 3 U/g·[dry cell weight] with p-Nitrophenyl caprylate (pNPC) as the substrate, which is 2 times as high as that of FLR. In addition, the cell-surface display systems based on FS or FL endowed the displayed RML with different enzymatic properties. The surface-displayed RML with its C-terminus free (FSR) showed a better catalytic performance at temperature, pH and thermostability than the surface-displayed RML with its N-terminus free (FLR) did. The results suggest that the surface display of RML based on FS anchor system is more promising and more effective, especially for N-terminal immobilization of target enzyme whose catalytic site is near the C terminus.]]> 2009/11/6 0:00:00 HAN Shuang-Yan,HAN Zhen-Lin,LIN Ying and ZHENG Sui-Ping 693true 2009/12/15 0:00:00 LI Fei,BO Xiao-Chen,LI Peng,YU Zhao-Hui,PENG Yu-Xing and WANG Cheng-Qi 692true MRS2 encode a magnesium transporter protein in Saccharomyces cerevisiae and deletion of MRS2 gene results in decrease in mitochondrial magnesium concentration, groupⅡ RNA splicing defect and growth defect on nonfermentable carbon source. To obtain more information of magnesium homeostasis in mitochondria, mTn-lacZ/LEU2 transposon library was transformed into mrs2 deletion mutant to screen for suppressor genes of MRS2. YMR166C, a member of mitochondrial carrier family, was identified as a suppressor gene of MRS2. Deletion of YMR166C gene can rescue the defects of mrs2 deletion mutant such as the decrease in mitochondrial magnesium concentration, groupⅡ RNA splicing defect and growth defect on nonfermentable carbon source. For the first time it was demonstrated that YMR166C is involved in mitochondrial magnesium homeostasis.]]> 2009/11/24 0:00:00 WANG Juan,FAN Qiang-Wang and ZHOU Bing 691true mtm1 deletion mutant to screen for suppressor genes of MTM1. The damage caused by MTM1 deletion is irreversible and even overexpression of MTM1 can not rescue the growth defect of mtm1 deletion mutant. Another screening strategy was adopted: a plasmid overexpressing MTM1 was transformed into wild type before the MTM1 gene on chromosome was deleted. The resulting strain, designated YES2MTM1, was transformed with a yeast genomic library. Transformants lost the plasmid overexpressing MTM1 after 5-FOA treatment. Yeast strains able to grow on nonfermentable carbon source with MTM1 deletion and overexpression of some DNA fragments were picked up and candidate suppressor genes were identified. Overexpression of five genes were identified to be able to rescue the growth defect on nonfermentable carbon source. The study will provide reference for MTM1 gene function and screening for suppressor of genes whose deletion result in irreversible damage.]]> 2009/10/28 0:00:00 WANG Juan,ZHANG Ming-Jie,ZENG Ya-Xue,CAI Ying and ZHOU Bing 690true in vitro and in vivo ubiquitination essays. These findings suggest that OGCP may be one of the ubiquitination substrate proteins of Parkin protein and Parkin protein mediates can promote the OGCP’s ubiquitination.]]> 2009/11/30 0:00:00 WANG Chun-Yu,CAO Li,TANG Bei-Sha,ZHANG Hai-Nan,GUO Ji-Feng,HE Dan,LIAO Shu-Sheng,TANG Jian-Guang,YAN Xin-Xiang and TAN Li-Ming 689true E. coli. Expression of rUCP1 can lower host’s growth rate. Immuno-electron microscopy proved expressed rUCP1 was mainly located on membrane. Purified rUCP1 was reconstituted into liposome and exhibited proton translocation activity. These results revealed eukaryotic UCP1 could be expressed in active form by prokaryotes and enable us to obtain enough amount of rUCP1 for structural study.]]> 2009/12/15 0:00:00 ZHOU Qiang-Jun,SUN Ji,ZHAI Yu-Jia and SUN Fei 688true MetaGen, a user-friendly utility to batch process and model the organism-specific multi-level metabolisms in KEGG into enzyme and pathway graphs, was developed. An example was given by expanding a little on the bow-tie structure of metabolic networks to show MetaGen is a useful and promising tool. MetaGen takes advantages of KEGG web service to ensure data reliability, uses relational database to accelerate the modeling process and facilitate data management, and applies advanced software modeling techniques as well as a pluggable software architecture for the easy expansion of functionalities in the future. MetaGen saves researchers from the elaboration on preparing metabolic networks for computation, which paves the way to deepen the researches on the large-scale metabolic networks. MetaGen is fully open-sourced and available at http://bnct.sourceforge.net/.]]> 2009/9/30 0:00:00 ZHOU Ting-Ting,YUNG Kin-Fung,CHAN Chun-Chung Keith,WANG Zheng-Hua,ZHU Yun-Ping and HE Fu-Chu 687true 2009/11/24 0:00:00 GUO Xing-Yan,ZHU Jing,HU Xin-Tian,ZHOU Rong-Wu and LUO Ming-Yang 686true in vivo and in vitro, and phosphorylation immunoblotting showed that ERβ can be phosphorylated by c-abl. Furthermore, the luciferase assay showed that c-abl enhanced ERβ mediated transcriptional activities, while c-abl inhibitor STI571 abolished c-abl mediated upregulation of ERβ transcriptional activity. These data suggest that c-abl was a novel co-activator of ERβ, which may uncover a role of ERβ signaling in cardiovascular disease.]]> 2009/10/22 0:00:00 GAO Yuan,CHEB Xuan-Nan,WEI Cong-Wen,ZHENG Zi-Rui,MA Hong-Fang,NI Cai-Fei,SONG Ting,QI Gou-Xian,ZHONG Hui and HE Xiang 685true 2009/9/27 0:00:00 JIN Xiao-Dong,LI Qiang,LI Ping,WU Qing-Feng,TAO Jia-Jun,HAO Ji-Fang,DAI Zhong-Ying and LIU Xin-Guo 684true Neurospora circadian clock system. The result shows that the correlation time of colored noise can affect strongly the strength of ISSR. For the case of no external signal(ES), the correlation time of colored noise plays a suppressive role for ISSR, and the suppressive role is increased with the increment of correlation time of colored noise. When the ES is injected to the system, with the increasing of the correlation time of colored noise, not only its suppressive role for ISSR is increased, but also the single peak ISSR can be transformed as internal single stochastic bi-resonance(ISSBR). There exists an optimal frequency of ES for the ISSR information amplification, while the ISSR is suppressed for another frequency of ES. In contrast to external colored noise, internal colored noise is more efficient to sustain and amplify the ISSR information. Furthermore, there exists a critical noise intensity for destroying the difference between white and colored noises.]]> 2009/11/6 0:00:00 SHI Jian-Cheng and LUO Min 683true TCID50 is 7.5, HA titer maintain at 1∶256 and EID50 is 8 which was detected using SPF eggs. The stable of reassortant is determined by RT-PCR gene segments from virus which were propagated in eggs. The morphology of reassortant conform the wild type virus. In order to test the immunogenicity, the reassortant viruses were purified. Mice were intranasally immunized and intramuscular injected with inactive whole virus as control. The high HI titer can be detected in both groups, seems hinger in i.m.(intramuscular) immunized groups (P=0.044), but higher IgA titer can only detected in i.n.(intranasal) immunized group. Compared with i.m. immunized group, higer pro-inflammation cytokines IL-1β, TNFα, IFN-α were tested in i.n. groups, means faster and stronger mucosal immune response was induced. Virus load were detected in lung, brain, spleen 4 days after immunization to determine the safety of reassortant as vaccine candidate, no detectable virus were found. All results show that the vaccine candidate can be used for vaccine production.]]> 2010/10/12 0:00:00 DUAN Yue-Qiang,LUO De-Yan,XING Li,YANG Peng-Hui,ZHAO Zhong-Peng,JIA Wei-Hong,LI Pei-Feng and WANG Xi-Liang 682true 2010/9/21 0:00:00 PAN Chang-Jiang,DONG Yun-Xiao,NIE Yu-Dong and WANG Yuan-Liang 681true 2010/9/14 0:00:00 ZHOU Xiao-Hai,LIU Yu,HU Nai-Yue,WANG Chun-Yan and ZHAO Li-Qing 680true Pseudolarix kaemferi Gordon), exhibited cytotoxicity in many human tumor cell lines. High content analysis (HCA) is a fluorescence microscopy-based automated technology used for quantitative analysis of multiple targets in cells. HCA could yield rich information about the temporal-spatial dynamics of the fluorescence-labeled cell constituents. The mechanism of inhibitory effects of pseudolaric acid B on human breast cancer MCF-7 cells was explored by high content analysis and flow cytometry. As shown by sulforhodamine B assay, PAB inhibited the proliferation of MCF-7 cells in a dose-dependent and time-dependent manner, and the 50% inhibition concentration (IC50) for 72 h was (1.80±0.33) μmol/L. Flow cytometry (propidium iodide staining) showed that, after treatment with PAB for 24 h, the proportion of MCF-7 cells at G2/M phase could increase to about 93%. Flow cytometry (annexin V-FITC and propidium iodide staining) showed that, PAB induced apoptosis of MCF-7 cells. High content analysis showed that: after treatment with PAB for 16 h, the mitotic index of MCF-7 could increase to about 40%, and cyclin B1 was upregulated; PAB caused dose-dependent disassembly of microtubules and inhibited the formation of mitotic bipolar spindles; PAB induced increase of mitochondrial mass; PAB induced grape-like giant nuclei indicating mitotic slippage in MCF-7 cells. These results suggest that PAB inhibits MCF-7 cell proliferation and induces apoptosis, these inhibitory effects may be related to disassembly of microtubules, spindle abnormalities, mitotic arrest and increase of mitochondrial mass.]]> 2010/10/12 0:00:00 DUAN Shao-Wei,XU Bo,CHEN Yun-Li,LI Min,FU Hong-Zheng and CUI Jing-Rong 679true 2010/9/21 0:00:00 LI Hong-Xia,ZHAO Xing-Bo,XU Ning-Ying,JIANG Yun-Liang,CAO Meng,LIU Yu-Fang,FANG Chi and LI Ning 678true 2010/10/12 0:00:00 GUO Jian-Xiu,RAO Ni-Ni,LIU Guang-Xiong,LI Jie and WANG Yun-He 677true 2010/6/17 0:00:00 OUYANG Qing,HU Rui-Cheng,DAI Ai-Guo,TAN Shuang-Xiang,XIAO Zhi-Qiang and TANG Cen-E 676true Prunus salicina seed meal via enantioselective transcyanation of acetyltrimethylsilane with acetone cyanohydrin in the water/organic solvent biphasic system was studied by GC analysis on a chiral β-Cyclodextrin Column. The effects of different (R)-oxynitrilase sources, diameter size of crude enzyme, substrate concentration, the ratio of two substrates, enzyme concentration and substrate structure on the enzymatic enantioselective transcyanation were investigated systematically. The results showed that (R)-oxynitrilase from Prunus salicina could efficiently catalyze enantioselective transcyanation of acetyltrimethylsilane with acetone cyanohydrin. The optimal diameter size of crude enzyme, concentration of acetyltrimethylsilane, ratio of acetone cyanohydrin to acetyltrimethylsilane and crude enzyme concentration were 0.3～0.45 mm, 21 mmol/L, 2∶1 and 60.9 g/L, respectively. (R)-oxynitrilase from Prunus salicina could not accept 3, 3-dimethyl-2-butanone as substrate, while both high substrate conversion and high product ee% were obtained with its silicon counterpart acetyltrimethylsilane as the substrate. Under the optimal conditions, both acetyltrimethylsilane conversion and enantiomeric excess of the product were above 99%. These results demonstrated that the silicon atom in substrate served as a more effective atom than the carbon atom to enhance the activity of this enzyme.]]> 2010/8/3 0:00:00 LIU Sen-Lin and ZONG Min-Hua 675true Xenopus tropicalis is an important model animal in immunology. Based on the available genome data, the genomic organization of all three light chain gene (ρ, σ and typeⅢ) loci were recently characterized in Xenopus tropicalis. On the basis of similarity of protein sequences, genomic organization and chromosomal location of the light chain genes among frogs and mammals, the data strongly support the previous suggestions that the ρ genes belong to the κ gene lineage, whereas the typeⅢ genes share a common origin with the λ genes. About 200 cDNA fragments from each of ρ, σ and typeⅢ genes, were cloned and sequenced respectively generating 22, 25 and 44 clones that have unique VJ junctions (after removal of redundant clones). Recombined VJ junctions of cloned light chain cDNA were analyzed, which showed a paucity of N and P nucleotides in expressed ρ, σ and typeⅢ genes. They also show that somatic VJ rearrangements of the σ gene seemed to be dependent on short stretches of homologous nucleotides (microhomology). The microhomology-directed VJ recombination obviously results in very limited diversity. However, most expressed ρ and typeⅢ VJ junctions showed direct ligation or deletions of V and J coding ends. These results help researchers to understand the generation of Ig diversity in X. tropicalis.]]> 2010/6/17 0:00:00 QIN Tong,REN Li-Ming and MENG Qing-Yong 674true cis and the splicing can occur independently of cellular entities exhibiting a splicing activity in and out of the cell. In order to improve the efficacy of intein-based dual-vector delivery of the BDD-FⅧ gene, here an additional acidic region-3 (AR-3) between Pro1640 and Ser1690 of FⅧ proven to be helpful for the secretion of heavy chain was incorporated into BDD-FⅧ heavy chain to examine its effect on secretion and bioactivity of an intein-spliced BDD-FⅧ protein. By co-transfection of the cultured HEK293 cells with genes of the ar-3 incorporated heavy chain and light chain with fused intein (HCAR3IntN and IntCLC), an ELISA and Coatest assay were performed to determine the amount of spliced BDD-FⅧ protein and coagulation activity secreted in the culture supernatant, and the intracellular BDD-FⅧ splicing was observed by Western blotting. The data showed that the amount of spliced BDD-FⅧ protein (173±26) μg/L and coagulation activity (1.31±0.15) U/ml of supernatant from gene co-transfected cells were greater than supernatant from intein-fused ar-3-free heavy and light chain genes (HCIntN and IntCLC) co-transfected cells (102 ± 12) μg/L and (0.79 ± 0.09) U/ml indicating the additional AR-3 in the heavy chain could dramatically improve the secretion and activity of intein spliced BDD-FⅧ. A spliced BDD-FⅧ protein ((35±7) μg/L) and coagulation activity ((0.28±0.08) U/ml) was also detected in the culture supernatant of combined cells separately transfected with the HCAR3IntN and IntCLC genes implying the cellular entities independent BDD-FⅧ splicing of the intein. The total protein from gene co-transfected cells displayed an obvious protein band of the spliced BDD-FⅧ detected by a FⅧ polyclone antibody indicating the intracellular BDD-FⅧ splicing. It paved a way for ongoing study on protein trans-splicing based dual-adeno-associated virus (AAV) delivery of the split BDD-FⅧ gene in gene therapy for hemophilia A animals.]]> 2010/8/17 0:00:00 Zhu Fuxiang,Yang Shude,Liu Zelong,Miao Jing,Qu Huige and Chi Xiaoyan 673true -1) intragastrically daily for 70 days in rabbits resulted in a significant inhibition of endothelium-dependent relaxation (EDR), a decrease of endothelium nitric oxide synthase (eNOS) activity in isolated aorta and the changes of biochemical parameters in plasma, including a decrease of superoxide dismutase (SOD), PON1 and AChE activity and nitric oxide level and an increase of malondialdehyde (MDA) level. In addition, a direct exposure of rabbit aortic rings to paraoxon also inhibited EDR. These findings suggest that subchronic toxicity dose intragastrically trichlorfon or vessels to be exposed directly to paraoxon in vitro could induce dysfunction of vascular endothelium. The mechanisms may involve an inhibition of antioxidases and oxidative stress induced by OPs.]]> 2010/9/13 0:00:00 XIONG Xiao-Ming,DAI Wen,LI Peng,WU Shu-Jin,HU Min and LIU Li-Ying 672true 2+ ACSF-perfused SD rat hippocampal slice, epileptiform discharges originated from CA3b and propagated to CA3c and CA1 region respectively.]]> 2010/8/31 0:00:00 GONG Xin-Wei,YANG Fan,LIU Jian-Sheng,LU Qin-Chi,GONG Hai-Qing,LIANG Pei-Ji and ZHANG Pu-Ming 671true 2010/8/3 0:00:00 LIU Min-Ji,WANG Xiao-Yan,SHEN Shou-Rong,LI Nan,ZHANG De-Cai,PENG Ya,GUO Qin and LI Gui-Yuan 670true in vitro and in vivo, and the effect of HIF-1α targeted RNA interference for reversing cisplatin resistance in human lung adenocarcinoma A549/CDDP. The HIF-1α shRNA was constructed and transfected into A549/CDDP and its xenograft animal model by pll-DCIONP and lipo2000, respectively. Fluorescent microscopy was employed to compare the transfection efficiency in vitro. The expression levels of HIF-1α, MRP1 and LRP after transfection were detected by reverse transcription polymerase chain reaction (RT-PCR) and immunocytochemistry analysis. Immunohistochemical analysis was performed to compare the levels of HIF-1α, MRP1 and LRP in transplanted tumors among different groups. In order to calculate the cisplatin resistant, MTT assay was performed to detect the cell half-maximum inhibitory concentration (IC50). The growth index of transplanted tumors after transfection were detected among groups. Additionally, HE staining of liver, kidney and brain tissues was used after magnetic iron oxide nanoparticles transfection. The method of pll-DCIONP was more efficient on transferring plasmid into cells than the lipids examined in vitro (P < 0.01). The mRNA and protein levels of HIF-1α, MRP1 and LRP of A549/CDDP were decreased after transfection with HIF-1α shRNA, and the resistance to cisplatin of A549/CDDP was reversed by 82%. The protein levels of HIF-1α, MRP1 and LRP in A549/CDDP transplanted tumors were decreased after transfection with HIF-1α shRNA; also, the growth of A549/CDDP transplanted tumors were inhibited by HIF-1α shRNA, cooperating with the synergistic effect of cisplatin. No necrosis of liver, kidney and brain tissue were observed after magnetic iron oxide nanoparticles transfection. The pll-DCIONP could be used as one of the ideal gene carriers for HIF-1α shRNA gene delivery in vitro and in vivo. HIF-1α can be an effective target for reversing cisplatin resistance in lung cancer, the mechanism underlying may be related to the decreased expression levels of HIF-1α, MRP1 and LRP after transfection with HIF-1α shRNA, and magnetic nanoparticle-mediated HIF-1α shRNA transfection has biological safety to some extent.]]> 2010/7/14 0:00:00 TU Xin,MIN Ling-Feng,CHEN Qiong,XIE Ming-Xuan and HE Ling-Ling 669true In vitro cell growth inhibition assay showed that anti-Trop-2 Fab could inhibit the Trop-2 positive cell BxPc3 growth, it illuminated that anti-Trop-2 Fab bound the Trop-2 on Trop-2 positive cell surface. For Trop-2 negative cell line NIH3T3, no significant inhibition among the different dosages of Fab. The results showed that anti-Trop-2 Fab antibody fragments could recognize Trop-2 extracellular domain in native conformation, making them as potential powerful reagents for clinical therapeutic application.]]> 2010/5/20 0:00:00 LIN Hong,LIANG Jie,ZHANG Hui-Lin,TANG Qi,SU Yi-Ping,CAO Brian,ZHU Jin and GUAN Xiao-Hong 668true 2010/6/10 0:00:00 YANG Yue,GONG Wei,WANG Wei-Wei,YU Xiao-Min,HU Song-Nian and YU Jun 667true ZmGT1, was cloned from maize (Zea mays) using RACE-PCR. The deduced ZmGT1 protein is highly homologous to glutathione transporters from other plants. A yeast mutant (hgt1Δ) deficient in glutathione transport was used to characterize the physiological functions of this gene. Complementation by ZmGT1 restored growth of the hgt1Δ mutant strain on a medium containing GSH as the sole sulfur source and also mediated the uptake of a model glutathione conjugate, GS-N-ethylmaleimide (GS-NEM). ZmGT1 was expressed in all organs of maize seedlings, with a higher level of transcript being found in leaves. The expression of ZmGT1 was strongly induced by atrazine with a 4～5 fold increase in transcript level being detected in leaves after 96 h treatment. The strong up-regulation of ZmGT1 by atrazine suggests that this glutathione transporter may be involved in the detoxification of xenobiotics.]]> 2010/6/23 0:00:00 PANG Sen,LI Xue-Feng,LIU Zhi-Qian and WANG Cheng-Ju 666true in vitro and in vivo assays. It was found that three dominant negative typeⅡ TGFβ receptors, which are dnBMPRⅡ, dnActRⅡ and dnActRⅡB, can not only reduce alkaline phosphatase (ALP) activity induced by BMP9 and calcium deposition, but also repress the activation of Smad signal pathway. Moreover, dnBMPRⅡ, dnActRⅡ and dnActRⅡB also showed to inhibit ectopic bone formation induced by BMP9 in vivo. However, target cells expressed BMPRⅡ and ActRⅡ, but not ActRⅡB. Then, when BMPRⅡ and ActRⅡ were silenced by RNA interference in target cells, luciferase reporter activity and ALP activity induced by BMP9 was accordingly inhibited along with knockdown of BMPRⅡ and ActRⅡ. Taken together, those results intensively suggest that BMPRⅡ and ActRⅡ are the functional typeⅡ TGFβ receptors required for BMP9 induced osteogenesis.]]> 2010/7/20 0:00:00 ZhAO Ying-Ze,ZHANG Yan and LUO Jin-Yong 665true n=3, P < 0.01), and over-expression of PIG11 induces translocation of Bax protein from cytoplasm to mitochondria in HepG2 cells. These results suggested that PIG11 protein can combine with Hsp60 in HepG2 cells, and the over-expression of PIG11 induce HepG2 cells apoptosis through mitochondrial pathway and the translocation of Bcl-2 family proteins Bax from cytoplasm to mitochondria maybe play an important role in the process.]]> 2010/8/17 0:00:00 LIU Xiao-Min,HU Rong,LIANG Xiao-Qiu,WANG Xiao-Juan,WANG Yan and ZHANG Yang 664true 2011/4/29 0:00:00 BIAN Cheng-Feng,ZHANG Ying,LI De-Feng and WANG Da-Cheng 663true 2011/7/13 0:00:00 PENG Zhi,JI Zhao-Ning,SHI Tai-Ping and MA Da-Long 662true 2011/3/23 0:00:00 LI Yu-Hui and LI Bing 661true P < 0.05) compared to control for rAAV-IRES-GFP (AAV-GFP) treated group. No effect on HDL-cholesterol concentrations occurred. Ultrasound determined intima-media thickness also has been significantly reduced compared to control. Serum hs-CRP and SOD levels increased significantly (P < 0.01). Serum MDA levels decreased significantly. Gene mRNA expression was detected in atherosclerosis rats' model. The results show that rAAV-hapoAⅠ-IRES-hSR-BⅠ vector can anti-inflammatory, reduce atherosclerotic macrophage content and increases lesion stability of pre-existing plaques through quenching of NF-κB activity and reducing plasma cholesterol. Simultaneous over-expression of apoAⅠ and SR-BⅠ by AAV-mediated gene transfer may have a favorable effect on diet-induced hypercholesterolemia and arteriosclerosis in rats. These results may provide a new method for gene therapy of arteriosclerosis.]]> 2011/3/29 0:00:00 LI Bing-Nan,LI Zhi-Yan,ZHANG Juan,LIU Zhen-Qing and TAN Meng-Qun 660true 2011/4/28 0:00:00 FAN Peng-Ju,YANG Xing-Hua,XIAO Mu-Zhang,LONG Jian-Hong and LEI Shao-Rong 659true via upregulation of phosphorylation of β-catenin and proliferation by the nuclear translocation of β-catenin.]]> 2011/4/11 0:00:00 ZUO Jian-Hong,LI Xin-Hui,LI Mao-Yu,WAN Xun-Xun,ZENG Gu-Qing,HE Qiu-Yan,LI Jian-Huang and XIAO Zhi-Qiang 658true 2011/4/13 0:00:00 ZOU Guan-Yu,JI Chao-Neng,SHEN Qi,XUE Jing-Lun and CHEN Jin-Zhong 657true cis-acting elements. The conserved region from -180 to +235 bp was found in the potential promoter among 6 vertebrate species by the ECR browser, and there have several potential binding sites for transcription factors, such as CCAAT/NFY, STAT1 and SP1. To characterize the NPCEDRG core promoter, transient luciferase and/or EGFP reporter assay were carried out with the construct pGL3-en138. The results demonstrated that the core promoter is located at the conserved region from -146 to -8 nucleotides. Gel shift assay revealed the specific binding of some nuclear proteins to probes containing a putative CCAAT/NFY site，suggesting that the CCAAT/NFY site contributes to the regulation of NPCEDRG gene expression.]]> 2011/4/13 0:00:00 HOU De-Fu,GUAN Yong-Jun,GUAN Rui,OUYANG Yong-Mei,YU Yan-Hui and CHEN Zhu-Chu 656true IL-28A gene was inserted into the insect cells expression vector pIZT-V5-His to generate recombinant vector pIZT/V5-His-hIL-28A, the vector was transferred into silkworm eggs by sperm mediated gene transfer, screening for gfp gene and verified by PCR and Dot blot hybridization. Transgenic silkworms were obtained after a specific band with the molecular mass of 25 ku could be detected in transgenic silkworm by Western blotting using an goat anti-hIL-28A antibody, and the content of hIL-28A in the G3 generation transgenic silkworms estimated by ELISA was approximately 0.198, 0.32, 0.238 ng/g in freeze-dried whole bodies, posterior silk glands and fat bodies, respectively. These results suggested that a heterologous gene could be integrated into silkworm genome by non-transposon vector and expressed successfully.]]> 2011/3/17 0:00:00 CUI Lin-Lin,XUE Ren-Yu,LU Ye,CAO Guang-Li and GONG Cheng-Liang 655true 2011/5/24 0:00:00 YANG Gen-Ling,LUO Wen-Ping,LIU Zhen-Zhen,TAN Dong-Mei,WANG Ying-Xiong and TAN Yi 654true 2011/3/23 0:00:00 LIU Chun-Liang,LIN Dan-Dan,XU Lan,JIANG Zhi and WU Shi-Liang 653true 2011/3/17 0:00:00 LIANG Jing-Wen,WANG Peng,CHEN Li,HU Yi-Qing and SUN Yi 652true Ustilago maydis CYP51 (P450-14DM, UmCYP51) protein in E. coli BL21(DE3), the different lengths of UmCYP51 gene that lacked the coding region for the putative membrane-spanning segment of the N-terminus were truncated. The first one is the wild type, the second one with 20 amino acids (60 base pairs) in N-terminus was truncated and the third one with 35 amino acids (105 base pairs) was truncated. Then these genes were incorporated into different expression vectors (pET28, pET32 and pGEX-KG) to construct nine recombinant expression plasmids (pET28-Um, pET28-Um-20, pET28-Um-35,pET32-Um, pET32-Um-20, pET32-Um-35, pGEXKG-Um, pGEXKG-Um-20 and pGEXKG-Um-35). The expression of recombinant plasmids were performed using 0.5 mmol/L of isopropyl β-D-thiogalactoside (IPTG) at 30℃. The culture harvested every 2 h up to 8 h. It was found that only recombinant plasmid pET32-Um-35 was expressed in E. coli BL21(DE3). Codon usage database (http//:www.kazusa.or.jp/coden) was used for the analysis of rare codon and software RNAStructure 4.5 was employed to study the mRNA secondary structure of translation initiation region. The results showed that rare codons rate in UmCYP51 gene is only 4.63%, the Rosetta (DE3) strain expressing some rare codons is not suitable for the protein expression of UmCYP51. Only the lowest energy of mRNA structure for pET32-Um-35 can obtained protein expression. These results are compatible with the experiments. Moreover, to design novel antifungal compounds against UmCYP51, based on the recently determined X-ray crystal structure human CYP51, a three-dimensional structure model of UmCYP51 was built through homology modeling using MODELLER 9V7 program. After refinement of the energy minimization and MD simulation using GROMACS 4.0.3 package, the UmCYP51 model was evaluated by PROCHECK Ramachandran plot statistics that indicated the designed model was in good quality. Commercial fungicide tebuconazole was docked into the model protein using Autodock 4.2.3 program to form the binding pattern of inhibitor with UmCYP51. The docking conformation of tebuconazole in the active site of UmCYP51 showed that the N-4 of the triazole ring was bound to heme iron with a distance about 0.245 nm. The hydroxy group of tebuconazole formed hydrogen-bonding interaction with the oxygen atom of carbonyl group for Ala265 with a distance about 0.245 nm. The mechanism of inhibitory activity of tebuconazole against UmCYP51 obtained from this study could aid in designing new antifungal compounds targeting this enzyme.]]> 2011/4/2 0:00:00 LI Shu-Xiang,HAN Rui,YUAN Li-Ling,XIONG Li,YUAN Yong-Ze,YANG Jiang-Ke,YAN Yun-Jun and LIU De-Li 651true 2011/4/11 0:00:00 SU Jian-Sheng,HAN Wen-fei,YAN Li,CHEN Li-Li,GU Jia-Mei,JI Ting-Ting and MENG Bo 650true 2+ current were shown after the stimulation. Activated astrocytes have an inhibitory effect on filopodia motility similar to extracellularly administrated ATP and glutamate. This indicates that activated astrocytes could probably inhibit dendritic filopodia motility through releasing ATP or glutamate. A method of selectively activating astrocytes in the mix culture of neuron and astrocyte was established, which provides a novel way for studying astrocytic signal regulation of neuronal development.]]> 2011/6/16 0:00:00 LIU Pu-Qing and ZHU Ke-Qing 649true in vivo and in vitro. However, relevant underlying molecular mechanisms of MR1 in AngⅡ induced cardiac hypertrophy remain to be elucidated. Gene silencing of MR1 by adenovirus-delivered siRNA approach in mice was used, and microarray analysis of myocardial gene expressions was compared before and after MR1 silencing along with AngⅡ treatment. Significant changes of genes expression in several pathways, such as pathways involved in cardiac hypertrophy and mTOR signaling etc., were observed between the two groups. Furthermore, genes that were changed by 10 folds after MR1 silencing were totally listed, with 39 genes being down-regulated and 5 up-regulated. To further verify the microarray results, some of genes that are suggested to be closely related to cardiac hypertrophy were detected by quantitative RT-PCR. As a result, HSP72 and thioredoxin 1 (Trx1) expression were increased, while the calcineurin Phosphatase β(CnAβ) and β-myosin (β-myosin) expression were suppressed upon MR1 silencing. These signaling pathways and genes are closely correlated with cardiac hypertrophy induced by Ang Ⅱ. The alterations of relevant pathways and genes may help understand the molecular role of MR1 in AngⅡ-induced cardiac hypertrophy.]]> 2011/4/28 0:00:00 DAI Wen-Jian,ZHANG Man,CHEN Jin-Jing,WANG Yi-Guang,KONG Wei-Jia and WANG Zhen 648true 2011/4/29 0:00:00 WU Wen-Qi,ZHENG Xiao-Bin,LIU Yong-Chu,TANG Kai and ZHU Huai-Qiu 647true 2011/4/11 0:00:00 LAN Jin-Ping,LI Li-Yun,JIA Lin,CAO Ying-Hao,BAI Hui,CHEN Hao,LIU Sheng-Nan,WU Lin and LIU Guo-Zhen 646true P < 0.05) and greater activation of its downstream mediators Smad2/3 during storage of FFP (P < 0.05). Both Day 0 FFP and Day 5 FFP stimulated EC migration in vitro; however, the effect of Day 5 FFP was significantly reduced. Inhibition of TGF-β typeⅠ receptor blocked FFP-induced Smad3 signaling in EC cells and restored the effectiveness of Day 5 FFP on EC migration to a comparable level as Day 0 FFP. These data suggest that the increased TGF-β levels during FFP storage contributes to the deterioration of stored FFP's effects on EC migration. A novel molecular mechanism contributing to the reduced efficacy of stored FFP was identified.]]> 2011/4/11 0:00:00 ZHANG Chun-Fang,TAN Tan,LI Jian-Guo,MA Hong-Ying,CHEN Zhu-Chu and DUAN Chao-Jun 645true E. coli together. The recombinant bacterial ghosts were then enfolded the Helicobacter DNA vaccine pcDNAKAT by incubating at 24℃ for 30 min. It was confirmed by DNA agarose electrophoresis,fluorescent microscope that plasmids pcDNAKAT could bound to bacterial ghost unspecificly, successful loading also resulted in the fluorescence shift indicated by the MFI of flow cytometric analysis. The anti-katA antibody titer of mice immunized with packaged DNA vaccine was 1∶(520 ± 54), which was significantly higher than that of control groups(P = 0.0058), which was indicated strongly that bacterial ghosts could deliver DNA vaccine effectively.]]> 2011/3/23 0:00:00 ZHANG Rui-Ping,LIU Chun-Jie,TAO Hao-Xia,LIU Xiang-Xin and ZHANG Zhao-Shan 644true 2011/4/11 0:00:00 Matloob Ammar,PAN Li-Na,PAN Hong,LU Jun and HUANG Bai-Qu 643true 2011/3/29 0:00:00 ZHANG Yong,ZU Xu-Yu,LUO Wei-Shi,TANG Sheng-Song and JIANG Yu-Yang 642true 2011/3/23 0:00:00 LIU Hong-De and SUN Xiao 641true -6 mol/L can predominantly promote ROB differentiation and biomineralization.]]> 2011/3/17 0:00:00 MING Lei-Guo,CHEN Ke-Ming,GE Bao-Feng and ZHAI Yuan-Kun 640true 2011/3/8 0:00:00 ZHU Yi-Min,XU Qi,DONG Lai-Rong and LIU Zhi-Ming 639true P <0.01). The expression of ADAM19 protein in the plasma of the normal group was (4.52 ± 0.10) μg/L, while that of the late-onset preeclampsia group was (4.32± 0.11) μg/L and that of the early-onset group was (3.78 ± 0.10) μg/L. There was significant difference between the early-onset group and the control group (P < 0.001), also between the early-onset group and the late-onset group (P < 0.001), while there was no significant difference between the late-onset and the control group (P > 0.05). The over-expression of ADAM19 in the placenta of preeclampsia patients may be associated with the occurrence and development of preeclampsia, and ADAM19 may be a marker used for predicting preeclampsia.]]> 2011/3/17 0:00:00 YANG Ling-Ling,WANG Jing,YUAN Peng-Bo,ZHAO Yang-Yu and WANG Yong-Qing 638true Arabidopsis thaliana (Col-0), treated at 4℃ for 4 h and 24 h together with their 24 h recoveries at 20℃. Co-immunoprecipitation coupled with sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis and MALDI-TOF MS identification was used to explore Rubisco-interacting proteins. Five protein candidates were profiled. The identified AAA-type ATPase family and glycosyltransferase were determined crucial for Rubisco activity. It is also strongly correlated to cold acclimation. Results suggest that the disassembly of Rubisco might have been the main cause of photosynthesis rate reduction under chill conditions, rather than photosystem or biogenesis involvement.]]> 2011/3/23 0:00:00 AN Bai-Yi,LIU Xiao-Yu,TAN Hua,LIN Wei-Hong and SUN Li-Wen 637true in vitro. In summary, it was demonstrated that both syntenin and Pick1 were expressed during RANKL-induced osteoclast differentiation of RAW264.7 cells, and EphB4/ephrinB2 reverse signaling regulates the expression levels of Pick1, but not of syntenin. These data help to preliminarily explore the potential PDZ-domain proteins involved in the downstream of ephrinB2 during the osteoclast differentiation of RAW264.7 cells in vitro.]]> 2011/3/17 0:00:00 MAO Ying-jie,HUANG Xu,ZHAO Juan,SHI Yue-Hua and GU Zhi-Yuan 636true 2011/1/31 0:00:00 XUE Yan,LIU Lin-Lin,HE Fen,JIANG Xue,ZENG Xian-Lu and BA Xue-Qing 635true 2011/2/24 0:00:00 ZHOU Hao,CHEN Zhuan-Hua,XING Zeng-Shu,WANG Gang,LUO Zhi-Gang,QIN Guo-Qing,LI Jian-Jun,QIAN Kun,JIANG Hong-Tao,WANG Fang,WANG Yi and LIAO Duan-Fang 634true ALP, osterix gene expression and mineralized bone modulus. After treatment of SEMF, the NOS activity in SEMF group increased in comparison with the normal control group (P < 0.01), reaching the highest level after 0.5 h. The gene expression of Osterix, ALPase activity and mineralized bone modulus in the SEMF group were also increased significantly. However, these effects were partially blocked in the L-NAME group．Surprisingly, all the osteogenic markers in the SEMF+L-NAME group were slightly higher than those of the control group, but lower than those of the SEMF group．In conclusion, The NO signal pathway was activated by SEMF treatment．The stimulatory effects of SEMF on the differentiation and mineralization of osteoblast were attenuated when NO signal pathway was blocked.]]> 2011/3/8 0:00:00 CHENG Guo-Zheng,GE Bao-Feng,CHEN Ke-Ming,LI Zhi-Feng,ZHOU Jian and WEI Zhe 633true via molecular docking method in previous work. The key residues of IN_L708, 906 complex model were detailedly analyzed from the three perspectives (i.e. distance, energy and hydrogen bond). The results show that the complex model is similar to the IN_5CITEP complex structure from proteins data bank. The difference of the motion modes and correlativity between IN_L708, 906 model and IN monomer was investigated with principal component analysis and dynamical cross-correlation map methods. The computational results indicate that the association with L708, 906 leads to the flexibility decrease of functional loop region, the lose of regular motion, as well as the disordered increase of correlative motion, which may be the main reasons for the activity attenuation of IN. All the simulation results may help the anti-HIV drug design based on the structures of Aryl diketoacids.]]> 2011/1/28 0:00:00 HU Jian-Ping,LIU Wei,Tang Dian-Yong,Zhang Yuan-Qin and Chang Shan 632true 2011/2/24 0:00:00 WEI Xiao-Lan 631true Spirulina platensis. The antihypertensive effect of VEP in spontaneously hypertensive rats (SHRs) was investigated in 24 h after one single dose and in one-week with one single dose per day. The expression regulation of VEP on major components of the renin-angiotensin system (RAS) in the kidney and serum of the SHRs was also explored with Real-Time PCR and enzyme-linked immunosorbent assay (ELISA). The results indicated that the least effective dose of VEP was 5 mg/kg and it exhibited a dose-dependent manner with increased dosages. The lowest weighted systolic blood pressure (WSBP) and weighted diastolic blood pressure (WDBP) occurred in 6 h and 4 h after administration, respectively. During the one-week experiment course, the WSBP of the VEP-treated group (10 mg/kg) was significantly lower than that of the negative control group from the 5th day. Furthermore, the VEP treatment significantly down-regulated the mRNA expression of renin, ACE, and the angiotensinⅡ(AngⅡ) type 1 (AT1) receptor, and up-regulated the mRNA expression of the AngⅡ type 2 (AT2) receptor in the kidney of the SHRs, suggesting that the antihypertensive effect of VEP might be related to its inhibition on the RAS and that it might be of great prospects in prevention and treatment of hypertension.]]> 2011/3/8 0:00:00 LU Jun,YANG Yang,CHEN Lin,REN Di-Feng,CAI Mu-Yi,WANG Jian-Zhong,EGASHIRA Yukari and TANOKURA Masaru 630true 2011/2/24 0:00:00 SHI Lei,JIA Ya-Li,ZHANG Xiao-Mei,YUAN Hong-Feng,ZENG Quan,ZHOU Jun-Nian,YUE Wen and PEI Xue-Tao 629true 2010/12/31 0:00:00 FANG Xin-Kui,WANG Xin and SUN Tao 628true in vivo, but the critical functional step of AFP is still obscurity. In order to explore AFP influences on the transduction of PTEN/AKT signal in HCC, expression of PTEN in human hepatoma cells and the effect of all trans retinoic acid (ATRA) on PTEN expression in these cancer cells were detected by Western blotting. AFP interacted with PTEN was analyzed by co-immunoprecipitation (Co-IP). Laser confocal microscopy was used to observe co-localization of AFP and PTEN. Short small RNA interfering (RNAi) was applied to knockdown the expression of AFP in Bel 7402 cells, and the phosphorylation of protein kinase B(AKT) was detected by Western blotting. Growth of Bel 7402 and HLE cells were assayed by MTT. Results showed that Bel 7402 cells expressed PTEN, and the expression of PTEN was induced by ATRA(160 μmol/L) mildly. Co-IP indicated that AFP has a property to interact with PTEN. PTEN co-localization with AFP was observed in cytoplasm of Bel 7402 cells. Constructed RNAi vector could knockdown the expression of AFP, expression of PTEN was promoted and phosphorylation of AKT was decreased when the expression of AFP was interfered or the cells were treated with ATRA(160 μmol/L). AFP-expressed vector pcDNA3.1-afp was transfected into human hepatoma HLE cells (AFP-non production). Co-IP analysis indicated that AFP interacted with PTEN, and expression of p-AKT(Ser473) was promoted in the tumor cells. It was also proved that pcDNA3.1-afp was able to reduce the role of Ly294002 in inhibiting the activity of AKT in HLE cells. These data provide the first evidence that AFP has a capacity to both interact with and inhibit activity of PTEN, which is also the pivotal events in the process that AFP activated the transduction of PI3K/AKT signal in hepatoma cells. Cytoplasmic AFP plays importance role on the resistance to ATRA induced apoptosis of HCC.]]> 2010/12/27 0:00:00 ZHU Ming-Yue,FU Shi-Gan,LI Meng-Sen,XIE Xie-Ju and LI Gang 627true 2+-binding motif is a stress protein highly expressed in the human esophageal squamous epithelial cells. It is downregulated in esophageal squamous cell carcinoma and functions as a modifier of deoxycholic acid (DCA) mediated cell injury. The S100 domain may be central to the function of CRNN. To further characterize the S100 domain of CRNN, the S100 domain in Escherichia coli, was cloned, expressed, purified and demonstrated that it was properly folded and suitable for biochemical and biophysical studies. More importantly, by nuclear magnetic resonance, gel-filtration, analytical ultracentrifugation, electrospray ionization-mass spectrometry, and Cross-linking analyses, a Ca2+-dependent multimeric property of S100 domain was identified. Furthermore, in response to DCA and ethanol challenge, the multimers have stronger protective effects on cells than dimers do. These data indicate that the S100 domain is a key domain in CRNN, which functions as a survival factor through multimerization. This work helps to further understand the feature of S100 domain and its association with cell injury.]]> 2010/11/23 0:00:00 LI Guo-Ming,CHEN Qiang,DAI Ke-Sheng and ZHENG Xiao-Feng 626true Δ1～42AA were respectively transfected into CNE2 cells by liposome-mediated transfection. Therefore, the CNE2/pcDNA3.1(+)- STGC3 and CNE2/pcDNA3.1(+)-STGC3Δ1～42AA cell lines of stable expression STGC3 were established. Here, it is reported that deletion of LG domain in this STGC3 reduces the tumor suppression activity of it, as demonstrated by drawing growth curve, experimenting plate clone formation as well as detecting cell cycle distribution. The results showed: the ability of STGC3Δ1～42AA suppressing CNE2 cell proliferation was obviously decreased to compare with wild type STGC3 (n=4, P < 0.05). These results indicate that the LG domain is necessary for the tumor suppression activity of the STGC3.]]> 2010/12/27 0:00:00 LI Li,HE Xiu-Sheng,LUO Qiao,ZHANG Zhi-Wei,YAO Xu-Jiong,CHEN Su-Qiong,LI Chun-Cheng,WANG Li-Li,DUAN Rong and CHEN Zhu-Chu 625true Laggera pterodonta is known to have an antiproliferative effects in vitro on human cancer. However, the exact mechanisms retain unclear. HTMF was investigated for its antiproliferative effects on human nasopharygeal carcinoma CNE cells. 3-(4, 5-Dimethylthiazol-2-yl)- 2, 5-diphenyl tetrazolium bromide (MTT) assay was used to deserve the inhibitory effect of HTMF. The changes of the cell and nuclear morphological characteristics were observed under the inverted and fluorescence microscope. The cell apoptosis was displayed by Hoechst 33258 staining and flow cytometry (FCM). The expression of Caspase3 and Caspase9 was detected by Western blotting. The mitochondrial membrane potential were analyzed by FCM and laser confocal microscope with Jc-1 fluorescence staining. MTT assay results show that HTMF significantly inhibited the growth of CNE cells in dose and time dependent manners. The IC50 values of HTMF were 69.02, 28.31 and 3.95 mg/L at 24, 48 and 72 h treatments, respectively. The apoptosis percentage in CNE cells is significantly increased compaired with control group. HTMF with 0, 5.0, 10.0, 20.0 and 40.0 mg/L at 48 h and 40.0 mg/L at 0, 6, 12, 24 and 48 h treatments, respectively increased the expression of Caspase3 and Caspase9 and degraded mitochondrial membrane potential in dose and time dependent manners. The mechanistic investigation revealed that HTMF has high inhibitory effects on the proliferation of CNE cells and induced the apoptosis of CNE cells by the decrease in mitochondrial membrane potential and increasing the expression of Caspase3 and Caspase9.]]> 2010/12/27 0:00:00 CAO Chang-Shu,SHEN Wei-Zai,LI Yao-Lan,WANG Hui and GAO Ming-Ya 624true 2010/11/18 0:00:00 ZHENG Jing-Min,ZHU Xiao-Dong,ZHANG Ming-Chao,WANG Jian-Ping,XU Li-Li and LIU Zhi-Hong 623true 2 (125 μmol/L) for 6 h, 12 h, 24 h, 48 h or 72 h respectively in vitro. RMCs cultures were maintained without CoCl2 in normal control group. The expression of GLAST and GS was determined by RT-PCR, Western blotting and immunocytochemical staining. L-[3,4-3H]- glutamic acid uptake was used to quantify glutamate uptake function of RMCs. The apoptosis of RMCs was confirmed by annexin V-FITC/PI staining. In early-stage of CoCl2-induced hypoxia (treated with CoCl2 for 6 h, 12 h, 24 h or 48 h)，the expression of GLAST was up-regulated (P < 0.001) and reached the maximum when RMCs have been treated with CoCl2 for 12 h (P < 0.05). After RMCs having been treated with CoCl2 for 72 h, the expression of GLAST had no difference compared to normal control (P > 0.05). CoCl2-induced hypoxia (treated with CoCl2 for 6 h, 12 h, 24 h, 48 h or 72 h) also up-regulated the expression of GS (P < 0.001) which reached the maximum when RMCs have been treated with CoCl2 for 48h (P < 0.001). The L-[3,4-3H]-glutamic acid uptake of hypoxia groups were the higher than normal control group (P < 0.05), and after having treated RMCs with CoCl2 for 48 h, the L-[3,4-3H]-glutamic acid uptake was the highest (P < 0.005). Treatments with CoCl2 did not induce apoptosis in RMCs. In early hypoxia stage, GLAST and GS were up-regulated and extracellular glutamate uptake increased. However, continued hypoxia causes gradual dysfunction and reduction of GLAST and GS, as well as glutamate uptake.]]> 2010/11/18 0:00:00 DAI Min and XIA Xiao-Bo 622true 2010/11/5 0:00:00 GAO Fei,SUN Xin-De and ZHANG Ji-Ping 621true Acidianus tengchongensis are hydrophilic.When these hydrophilic residues in ATcpnβ were mutated to hydrophobic residues, i.e. residues 236 from G to Y, 237 from M to F, 288 from D to L, 295 from A to V, 313 from A to V and 314 from K to V, they enhanced the inhibitory effect of ATcpnβ on the refolding of acid-denatured GFP and the inhibition activity of citrate synthase (CS) thermal aggregation. Hydrophobic interaction may contribute more to peptides binding affinity both in groupⅠ and groupⅡ chaperonin. Chaperonins have been proved to have ATP-dependent peptides refolding ability. However, it is still unclear whether peptides binding ability of chaperonins is ATP-dependent. Surface plasma resonance (SPR) analysis is used to test chaperonin binding ability to different peptides with different denaturing level. These assays revealed that ATcpnβ could capture guanidine hydrochloride denatured malate dehydrogenase in a Mg2+/ATP independent manner while it bind thermal aggregated citrate synthase or lysozyme in a Mg2+/ATP dependent manner. It has been proposed that chaperonin conformatinal changes induced by Mg2+/ATP to expose more hydrophobic surface is required for chaperonin capturing thermal aggregated peptides which has larger hydrophobic surfaces.]]> 2010/11/23 0:00:00 WANG Li,ZHANG Kai,FAN Zheng,DONG Zhi-Yang and SUN Fei 620true 2010/12/27 0:00:00 XUAN Bin,WANG Kai and ZHANG Da-Ren 619true MCC) of the 53 fold types were found to be 96.42%, 99.91% and 0.91, respectively. The result indicates that using the functional domain composition to represent a protein is very promising for protein fold recognition. And though based on simple classification rules, LIFCA can concentrate the functional features of proteins, reflecting the corresponding relation between structure and function.]]> 2010/11/10 0:00:00 YAN Jin-Li,CHEN Zhi-Wei,XU Hai-Song and LI Xiao-Qin 618true in vitro. However, there is nearly no reports on cell proliferation in vivo in plants under normal and stress conditions. Here, calmodulin antagonist W7 inhibited the root elongation of Arabidopsis seedlings, and the area and cell number of root apical meristem were reduced, the expression of CYCB1;1 gene which plays a key role in cell proliferation was seriously restrained. This data indicates that activated calmodulin is required for cell proliferation. Abscisic acid (ABA), as a key hormone mediating plant adaptation to various environmental challenges, regulates many processes of plant growth and development, such as seed germination and seedling growth. In the present of W7, wild type arabidopsis seedlings are insensitive to ABA, and cam2cam3cam4 triple mutant had decreased sensitvity to ABA. The results implied that CaM might play important role in cell proliferation of root apical meristem and involved in ABA response.]]> 2010/11/10 0:00:00 CHENG Yu-Dou,HAN Shu-Yun,ZHAO Jun-Feng,GAO Ying-Jie,SUN Da-Ye and CUI Su-Juan 617true 2010/4/20 0:00:00 Zhou Shou-Hong,YANG Xu-Hong,SONG Tao,WU Shu-Jin,HUANG Ning-jiang and LIU Li-Ying 616true 2010/11/23 0:00:00 CHEN Wei-Chun,SONG Jie,LIU Zhen-Jie,JIANG Zhi-Wen,YUAN Yuan,ZHENG Hui-Ling and LIU Xin-Guang 615true vp4, vp6 and vp7 were cloned from the rotavirus infected child stool specimen in Beijing through RT-PCR．The protein sequences and their antigenic determinants were predicted．According to the epitope peptide sequences, 4 epitopes from these structural proteins were chosen, a DNA fragment encoding all these epitopes was synthesized and cloned into the prokaryotic expression vector. Multiple epitope protein (rotavirus multiple epitopes, RME) expressed in E. coli can be recognized by the polyclonal antibody of rotavirus, and induce immune response in mice．The specific antibody IgG induced by RME can recognize human rotavirus (Wa strain), RME itself as well as individual epitope peptides. The antibody titer of IgG to RME is high (1∶40 000), while the titers to EV4, EV6 or EV7 are in a range of 1∶10 000 to 1∶20 000. However the IgG titer to the Wa strain is lower, i.e., 1∶2 500．Intriguingly, the RME-induced IgG can neutralize the Wa strain rotavirus challenge in the MAC145 cell line. This research has laid the foundations of producing effective bioengineering vaccines to rotavirus.]]> 2010/11/10 0:00:00 YANG Yan-Mei,LI Xia,YANG Hui,QIAN Yuan,ZHANG You,HUO Yan,FANG Rong-Xiang and CHEN Xiao-Ying 614true trans-splicing was explored to co-transfer split three fragmented vWF gene into eukaryotic cells by a ternary-vector system and the functional vWF protein was expected to be formed posttranslationally by protein trans-splicing. The vWF cDNA was broken into three fragments before codons of Cys1099 and Ser2004 which required for protein splicing and then fused with Ssp DnaE and Ssp DnaB inteins respectively. A group of three eukaryotic expression vectors were produced by inserting these three fusion genes into pcDNA3.1(+) respectively. By transient co-transfection of 293 cells with these three vectors the conditioned culture supernatant was observed for vWF multimer pattern by electrophoresis, analyzed for vWF antigen and binding capacity of coagulation factor Ⅷ (FⅧ) quantitatively. With FⅧ gene co-transfection, the antigen and activity of FⅧ in the culture supernatant were measured. The data showed that with intein-mediated protein trans-splicing after translation the culture supernatant from cells co-transfected with intein-fused three fragmented vWF genes displayed a vWF multimer pattern and FⅧ binding capacity similar to normal human plasma and cells of vWF gene transfected positive control, and the FⅧ secretion and activity were increased dramatically in FⅧ gene co-transfected cells indicating the functional recovery of spliced vWF as a FⅧ carrier. It suggests that inteins could be used as a powerful means for tri-fragmental co-delivery of the vWF gene and may be valuable for application of intein-based ternary AAV vector in split vWF gene delivery in gene therapy for VWD to overcome the packaging limitation of AAV vectors.]]> 2010/9/16 0:00:00 ZHU Fu-Xiang,YANG Shu-De,LIU Ze-Long,MIAO Jing,QU Hui-Ge and CHI Xiao-Yan 613true chs gene super-family, is a key enzyme in flavonoid biosynthesis. Two independent promoters was isolated for chsA, named PchsA-L (550 bp) and PchsA-S (354 bp) (GenBank accession number EF199747 and EF199748 respectively), from the genomic DNA of Petunia hybrida. PchsA-L differs with PchsA-S mainly in that PchsA-L has a 182 bp fragment from 88～269 bp, and the sequence 103～201 bp has the characteristics of a typical intron. Both promoter sequences contain conserved sequences of TATA box, CCAAT box, cap site (CCATAA), and the flower-specific promoter sequences TACPyAT box, anther box (TAGAAGTGACAGAAAT), G-box (CACGTG), box1 element (ATGTCACGTGCCATC) and box2 element (TGTGTTGAAGGTTTGCTA). The petunia plant used for promoter cloning was a diploid with 14 chromosomes. Southern blotting showed that both promoters had multiple copies in the genome. The two promoters segregated in the offspring but the segregation did not meet the ratio of 1∶2∶1. qRT-PCR analysis showed no significant difference in chsA gene expression in non-UV-treated and UV-treated floral organ of plants with one or both promoters. The expression of chsA in the UV-treated seedling leaves was increased compared with UV-treated floral organ, and PchsA-L-driven chsA expression in the UV-treated seedling leaves was very significantly increased than that driven by PchsA-S while there was no chsA gene expression in non-UV-treated seedling leaves. The results show the presence of two independent promoters PchsA-L and PchsA-S for chsA in the petunia genome; the 182 bp intron-like sequence in PchsA-L promoter could significantly increased chsA gene expression in UV-treated seedling leaves.]]> 2010/11/10 0:00:00 XIANG Tai-He,XU Ji-Ming,WANG Lin and LIN Lei 612true n = 228) composed of Wannan Spotted, Jixi Black and Dingyuan and LTG (n = 92) from Landrace, Large White, Duroc and their hybrid pigs and then detected the SNPs in the coding region of THRSP gene. The effects of SNPs on mRNA folding and protein secondary structure in FTPG and LTPG were predicted using Vienna RNA folding software and YASPIN web services, and their distribution and association with lipogenesis capability in FTG and LTG were analyzed. The results showed that the homologies of nucleotide sequence in coding region of pig THRSP gene with human and cattle were 86% and 88%, respectively. Two SNP sites, G123A and A308G, were located at 123 bp and 308 bp away from the start of the CDS, respectively. The mutation at G123A site was a synonymous mutation, but the mutation at A308G site caused the change of amino acids from lysine to arginine at 103rd site of THRSP protein which resulted in the changes of casein kinase Ⅱ phosphorylation site from TKEE to TREE, mRNA folding and protein secondary structure. The frequencies of alleles 123G and 308A were 0.975 9 and 0.589 9 in FTG, repectively, while that of alleles 123G and 308A were 0.657 7 and 0.815 2 in LTG. The total frequency of gamete 123G308A and 123G308G in FTG achieved 0.975 9, while that of gamete 123A308A and 123G308A in LTG was 0.815 3. Data from the present study imply the association of polymorphism at G123A and A308G sites with lipogenesis capability of pigs and their important regulating role in the expression of lipogenesis genes in pigs.]]> 2010/10/12 0:00:00 ZHOU Qian-Qian,CHEN Hong-Quan,WEI Han-qing,QIN Jie,CHEN Hua and ZHANG Yi-Peng 611true in vivo experiments demonstrated that overexpression of caveolin-1 resulted in significant growth inhibition of the xenograft pancreatic tumors. Immunohistochemistry analysis demonstrated both a marked decrease in the number of proliferating tumor cells and an increase in the number of apoptotic tumor cells in PANC1/cav-1 xenograft tumors. The results provide an initial demonstration that caveolin-1 can function as a tumor suppressor rather than as a tumor promoter in PANC1 cells.]]> 2011/6/17 0:00:00 WANG Xiao-Hui,ZHENG Ya-Min,CUI Ye-Qing,LIU Shuang,SUN Hai-Chen and LI Fei 610true TM, which can induce retroviruses release. Then we used the collected retroviruses to infect RAW264.7 cells and achieved adipophilin gene highexpression and knockdown RAW264.7 cell lines applying puromycin screening. After the infected RAW264.7 cells incubated with 50 mg/L Ox-LDL for 24 h, the lipids accumulation were measured by Oil red O staining and HPLC, the expression of mRNA and proteins of adipophilin and PPARγ were detected by semi-quantitative RT-PCR and Western blot respectively, and phosphorylation of ERK1/2 was analyzed by Western blot too. The results of enzyme-digesting confirmed the recombinant retroviral vectors pQCXIP-HA-Adipophilin and pSuper-retro-adipophilin siRNA as expected. In the situation of Dutch fat, transfected cells with pQCXIP-HA-Adipophilin significantly increased the accumulation of lipids, but reduced the expression of PPARγ and the phosphorylation of ERK1/2, which were reversed in cells with pSuper-retro-adipophilin siRNA transfection. Our results showed that ERK1/2 and PPARγ may be related with lipids accumulation caused by adipophilin expression in macrophages incubated with modified LDL. Therefore, adipophilin might contribute, in vivo, to lipid accumulation in the intima of the arterial wall through the ERK1/2-PPARγ signaling pathway.]]> 2011/7/6 0:00:00 LIU Qing-Nan,DAI Zhi-Bing,LIU Zhi-Qiang,TANG Chao-Ke,TIAN Guo-Ping,DAI Xiao-Song,HE Qiu,YE Ling and YUAN Zhong-Hua 609true 32P of the TNF-α promoter 62～85 bp could bind to nucleic proein STAT3, but not mutant 62～85 bp probe. These data suggested that the effect of STAT3 on TNF-α gene expression did not depend on LPS, and the potential binding site of STAT3 on TNF-α promoter might be between 80 and 85 base radical fragment.]]> 2011/8/24 0:00:00 YANG Li-Ping,YAO Yong-Ming,YE Qi-Nong and SHENG Zhi-Yong 608true 2011/11/14 0:00:00 PAN Wei-Nan,LI Feng,MAO Xiao-Huan,QIN Xu-Ping,DENG Shui-Xiu,FENG Fen,CHEN Feng,LI Lan-Fang,LIAO Duan-Fang and CHEN Lin-Xi 607true 2011/10/24 0:00:00 MAO Xiao-Huan,SU Tao,ZHANG Xian-Hui,LI Fang,QIN Xu-Ping,LIAO Duan-Fang,LI Lan-Fang and CHEN Lin-Xi 606true 2011/8/24 0:00:00 LI Xiang,LI Anan,GONG Ling,LIU Qing and XU Fu-Qiang 605true 2011/5/13 0:00:00 NI Bing,QIAN Chen-Can,WU Rui-Jie,ZHU Hong-Wei,LIU Zu-Xiang and LI Yong-Jie 604true IA was reported, and the rate of inhibition were (63.0 ± 2.2)% (1 mT), (55.0 ± 1.7)% (5 mT), (38.0 ± 1.8)% (10 mT), respectively. Moreover, the characteristics of activation and inactivation were both influenced by EMF, because there were a decreasing both on the half activation voltage and the half inactivation voltage. Additionally, different effects on the channels had been found for the different intensities. In the research, the maximum inhibition rate of current was induced by 1 mT, and the changing of the half activation voltage and the half inactivation voltage were the biggest after exposure to 5 mT, and 10 mT EMF increased the slope factor of inactivation. The results indicated that EMF affected the conformational changes of ion channel protein on cell membrane, and further influenced the normal function of ion channels.]]> 2011/6/17 0:00:00 Li Gang,LI Dan-Dan,LI Yuan-Yuan and LIN Ling 603true 2011/9/27 0:00:00 DENG Hui,ZHANG Yun-Li,ZHONG Bai-Yun,LIU Wei-Dong,YAN Qun,FENG Si-Si and XIAO Zhi-Qiang 602true 2O2. The models of apoptosis induced by H2O2 in HUVEC and the concentrations of TSG on apoptosis model were established via MTT, Hoechst33258 staining, and Flow Cytometry. The expression of Caspase-3 and PARP was detected by RT-PCR and Western blot. Compared with control group, the viability of cells is decreased and the apoptosis ratios is increased with increased concentration of H2O2(P < 0.01) respectively. And the group of 300 μmol/L H2O2 inhibited the cell proliferation, increased the number of apoptotic cells significantly. The viability of cells is increased and the apoptosis of HUVECs is decreased after pretreated with different concentration of TSG. According to the MTT and Flow Cytometry, the optimal concentration for H2O2 to establish apoptosis model and for TSG to protect HUVECs induced by H2O2 were 300 μmol/L and 10 μmol/L respectively. Compared with the control group, the group of 300 μmol/L H2O2 inhibited the cell proliferation, increased the number of apoptotic cells and the expression of Caspase-3 significantly. Compared with H2O2 group, 10 μmol/L of TSG improved the rate of cell proliferation, inhibited cell apoptosis, and decreased the expressions of the Caspase-3 and PARP significantly (P < 0.01). These results indicate that TSG can inhibit H2O2-induced apoptosis of human umbilical vein endothelial cells, and its mechanism may relate to the down-regulation of Caspase-3 and PARP expression.]]> 2011/7/13 0:00:00 LONG Shi-Yin,ZHANG Cai-Pin,QIAO Xin-Hui,HUANG Liang-Zhu,TIAN Ying,GAO Xi-Qiang and TONG Li 601true 642～1290 helicase by fluorescence polarized technology and free phosphorus assay technology; and the parameters of binding between LMX and helicase were studied by fluorescence and ultraviolet absorption spectroscopy, included binding constants, number of binding sites, the type of acting force, and binding distance. The results indicated that the reaction between the helicase and LMX was occurred spontaneously, there was one binding site between two molecules, the helicase and LMX might compound BLM-LMX complexes caused by electrostatic force and hydrophobic interaction force; moreover, the intrinsic fluorescence of the helicase was static quenched by LMX as a result of non-radioactive energy transfer. In this process, the helicase and ATPase activities were inhibited and DNA-binding activity of the helicase was promoted by LMX. The mechanism of effects of LMX on biological properties of BLM helicase may be included as below: LMX could inhibit the ATPase activity by allosteric mechanism and stabilize the conformation of the enzyme in low helicase activity state, destroy the coupling of ATP hydrolysis to unwinding, and inhibit the unwinding dsDNA by blocking helicase translocation. The reason that LMX could promote DNA-binding activity of the helicase may be the substitutional functional groups at C-6 and C-7 of LMX which may enhance enzymes activity and strengthen the attachment of drug-enzyme-DNA complex. The results may provide the relative theoretical basis for studying the molecular mechanism of DNA helicase as drug target and understanding the mechanism of action of quinolone drugs.]]> 2011/6/17 0:00:00 LUO Heng,CHEN Xiang,DING Mei,YANG Qi-Xin and XU Hou-Qiang 600true 2011/6/17 0:00:00 LV Da-Wei,CAI Xu and WANG Yan 599true 2011/6/17 0:00:00 XU Yu-Xia,WANG Hong-Quan,ZHAO Hong,GUO Jing-Chun and ZHU Cui-Qing 598true Tobacco necrosis virus A Chinese isolate (TNV-AC). TNV-AC-based virus induced gene silencing (VIGS) containing different structural arrangements of sequences targeting specific genes and variations in the positions of exogenous fragment insertions were evaluated along with temperature effects on inoculated plants. Replacement of the coat protein (CP) gene by exogenous gene fragments could not be applied to TNV-AC because the chimeric TNV-AC constructs were unable to induce silencing of corresponding endogenous gene and derivatives lacking the CP could not move systematically in Nicotiana benthamiana. We found that the optimal position for insertion of exogenous gene fragments was the region immediately behind the stop codon of the CP gene, and the maximum lengths of fragments that could be accommodated within virions was around 120 nt. An NbPDS fragment inserted into TNV-AC as an inverted repeat produced a VIGS derivative that could silence endogenous NbPDS in N. benthamiana with extremely high efficiency. We also found that temperature could significantly affect gene silencing efficiency of inoculated plants and the stability of heterologous gene fragments. Plants grown at 18℃ exhibited substantially higher gene silencing efficiency compared with 24℃, and the duration of silencing persisted for more than 110 days. The TNV-AC-based VIGS vectors were also able to induce efficient silencing of endogenous Su and ChlH genes in N. benthamiana. Taken together, our results show that TNV-AC has the potential for development into a novel VIGS vector suitable for functional genomics of N. benthamiana.]]> 2011/5/24 0:00:00 GAO Yang,ZHANG Yong-Liang,ZHANG Xiao-Feng,HAN Cheng-Gui,YU Jia-Lin and LI Da-Wei 597true in vitro microneutralization assay, furthermore, the anti-releasing effects of hemagglutination existed in the antisera. Alignment with databases showed that the amino acid residues of two epitope peptides are highly conserved amongst the NA sequences of the strains isolated from the world. These findings indicate that SE8 and RE6 represents an attractive candidate for an effective synthetic peptide-based vaccine against 2009 A H1N1 viruses.]]> 2011/5/16 0:00:00 HUANG Ping,XU Yuan-Sheng,ZHONG Jing,NI Han-Zhong,ZHANG Yong-Hui,CHEN Qiu-Xia,TAN Song-Nuan and LI Hui 596true in vitro and in vivo. In summary, we showed here that the promoter of MHV-68 ORF9 is responsive to RTA activation through ORF9p-RRE. The study provides support to further investigate and understand the role of RTA in regulating viral gene expression and viral life cycle.]]> 2011/5/16 0:00:00 HONG Yun,QI Jing,GONG Dan-Yang,DENG Jian-Wen and DENG Hong-Yu 595true 12 numbers used for ribosome display was constructed. Among the 180 selected clones, four clones RB24, RB71, RB109 and RB156 which exhibited the highest ELISA signals were isolated. The analysis of their sequences showed that they were new human immunoglobulin V genes to RVGp. And the reconstructed VH-Lc-VK antibody fragments can recognize RVGp specifically and antagonize the cytolytic effect of rabies virus on Vero cells. The prepared VH-Lc-VK fragments to RVGp will be useful for preparing engineering antibodies with high affinity against rabies virus. Ribosome display is a rapid means of generating fully human antibody fragments in vitro.]]> 2011/5/16 0:00:00 ZHAO Xiao-Ling,CHEN Wei-Qiang,FENG Hong,SHEN Cheng-Feng,JI Yan,LI Jing-Mei,ZHANG Su-Juan and YANG Zhi-Hua 594true 2011/5/16 0:00:00 SUN Guo-Peng,YE Xian-Long,REN Gui-Ping,LI Jin-Nan and LI De-Shan 593true 2011/4/2 0:00:00 YU Yun,LIN Ju-Qiang,HUANG Hao,CHEN Yan-Ping,FENG Shang-Yuan,SU Ying,CHEN Jie-Si and CHEN Rong 592true in vitro, the newborn rat calvarial osteoblasts were isolated by enzyme digestion and randomly divided into 5 groups after one passage．The EMFs treatment group is exposed to 50 Hz 1.8 mT sinusoidal, triangular, serrate and square wave of EMFs and controls without EMFs treatment. The osteoblasts were exposed to the EMFs for 30 min/(time·day). The cells were observed under the contrast phase microscope each day. The EMFs groups were arranged in spiral appearance after 4～8 days of the EMFs treatment. MTT result showed that sinusoidal wave significantly inhibited osteoblast proliferation, whereas square wave enhanced the osteoblast proliferation. The ALP activity in triangular and sinusoidal wave groups significantly increased (P < 0.01) compared with that in control. The number of calcified nodules, ALP stained result and Collagen-Ⅰimmunohistochemistry tested result are according to the result of ALP activity. There was a common result that sinusoidal and triangular wave significantly increased the Igf, Opg and Runx-2 mRNA expression. The four different wave of EMFs at 50 Hz 1.8 mT enhances the osteoblasts maturation, mineralization and Opg and Runx-2 mRNA expression of the osteoblasts, while sinusoidal and triangular have the strongest effect.]]> 2011/6/22 0:00:00 ZHOU Jian,MA Hui-Ping,CHEN Ke-Ming,GE Bao-Feng,CHENG Guo-Zheng,WANG Jia-Qi and WEI Zhe 591true Helicobacter pylori plays essential roles in the colonization of the bacteria in human hosts and is required for the utilization of heme as the sole iron source. Residue His245, which is highly conserved, coordinates the heme iron through its sidechain imidazole group. Surprisingly, this residue was not required for the enzymatic activities of HugZ. To investigate the roles played by His245 in heme binding and enzymatic mechanisms of HugZ, we have solved the crystal structure of HugZ mutant H245A at 2.55Å resolution and found that a nearby histidine residue, His249, coordinates the heme iron. This substitution is made possible by the fact that both residues 245 and 249 are located in a flexible loop region ranged from Gly239 to the C-terminus. Similar structural features have not been observed in other heme oxygenases so far. We have also performed spectroscopic studies on the heme-binding properties of HugZ and relevant mutants and our results suggest that the flexible C-terminal loop region of HugZ and the presence of multiple histidine residues in this region may play important roles in heme recruiting and in the catalytic mechanisms of HugZ.]]> 2011/12/20 0:00:00 SHEN Xi-Hui,HU Yong-Lin and WANG Da-Cheng 590true in vivo and in vitro. Overexpression of JNK3 inhibited NF-κB- dependent transcription induced by TNFα. It was demonstrated that JNK3 decreased NF-κB binding to its cognate DNA sequences and NF-κB target genes expression. Taken together, these data suggest that JNK3 may function in vivo as a modulator in suppressing the transcriptional activity of p65.]]> 2012/4/28 0:00:00 LI Qiang,HAN Qing,YU Dong-Hui,TANG Liu-Jun,WANG Jian,WANG Xiao-Hui,XU Wang-Xiang,ZHAN Yi-Qun,LI Chang-Yan,GE Chang-Hui,YU Miao and YANG Xiao-Ming 589true 2012/3/13 0:00:00 XIANG Bo,WANG Wei,LI Wen-Juan,TANG Ke,ZENG Zhao-Yang,LI Xiao-Ling and LI Gui-Yuan 588true n=8, in each group): group Ⅰ(5 ml 2, 6-diisopropylphenol was injected peritoneally in the Sprague-Dawley rats by dosage of 100 mg/kg); group Ⅱ(5 ml 2, 6- diisopropylphenol was injected peritoneally in the Sprague-Dawley rats by dosage of 100 mg/kg and giving a course of electroconvulsive shock); group Ⅲ(5ml Saline was injected peritoneally in the Sprague-Dawley rats); group Ⅳ(5 ml saline was injected peritoneally in the Sprague-Dawley rats and giving a course of electroconvulsive shock). The Morris water maze test started within 1 day after the course of electroconvulsive shock finished in order to evaluate learning-memory. The hippocampus was removed from rats within 1 day after the Morris water maze test finished. The content of glutamate in the hippocampus of rats was detected by high performance liquid chromatography. The content of Protein Tau which includes Tau-5 (Total protein Tau), p-PHF1Ser396/404, p-AT8Ser199/202, p-12E8Ser262, GSK-3β1H8 and PP-2A in the hippocampus of rats was detected with Western blotting. The electroconvulsive shock and the 2, 6-diisopropylphenol can induce the impairment of learning-memory in depressed rats, extending the evasive latency time and shortening the space exploration time. And both influences present subtract effect. The electroconvulsive shock can significantly up-regulate the content of glutamate in the hippocampus of depressed rats which was reduced by 2, 6-diisopropylphenol. And both influences present subtract effect. The electroconvulsive shock and the 2, 6-diisopropylphenol does not affect the total protein Tau and protein PP-2A in the hippocampus of rats. The electroconvulsive shock can up-regulate the hyperphosphorylation of protein Tau and the expression of GSK-3β1H8 in the hippocampus of depressed rats, which is reduced by 2, 6- diisopropylphenol. And both influences present subtract effect. Our results indicate that the electroconvulsive shock up-regulates the content of glutamate in the hippocampus of depressed rats, which up-regulates the hyperphosphorylation of protein Tau through up-regulating the expression of GSK-3β1H8, and further induce the impairment of learning-memory in depressed rats. Wheareas, the 2, 6-diisopropylphenol protects against the impairment of learning-memory and reduce the hyperphosphorylation of protein Tau induced by electroconvulsive shock in depressed rats through reducing the expression of GSK-3β1H8 and the content of glutamate in the hippocampus.]]> 2012/3/19 0:00:00 LIU Chao,MIN Su,WEI Ke,LIU Dong,DONG Jun,LUO Jie and LIU Xiao-Bin 587true Drosophila egg-laying behavior. We found that selection of egg-laying site in flies is a good model to investigate the food preference in Drosophila. On sugar food with lower concentration, flies showed significant preference that was reduced upon the inhibition of olfactory pathway. Moreover, to examine the plasticity of egg-laying behavior, a new learning paradigm was devised. We associated light-dark cycle of egg-laying with different food conditions and found that flies could maintain egg-laying memory at least 3 days after training. This egg-laying system provides the basis for further study of molecular and neural mechanism underlying the relationship between environment and behavior.]]> 2012/5/31 0:00:00 WANG Yi-Jin,WEN Sheng-Yun,GONG Hai-Yun,GONG Zhe-Feng and LIU Li 586true in vivo experiments. Furthermore, PROL4 transfected cancer cell strains were found to display an elevated sensitivity to cisplatin treatment than mock-transfectant control cells, and the sensitivity up-regulation was closely related to the rate of cell apoptosis induced by PROL4 and cisplatin cooperately. These results suggest that PROL4 is potentially a new genetic engineering target to enhance the chemotherapeutic effects of cisplatin in lung cancer treatment.]]> 2011/12/31 0:00:00 HE Jin-Jun,LIU Chuan-Jun,LIU Dan-Hui,LIU Ji-Fu,WU Shan-Shan,XIAO Xue-Yuan and HE Da-Cheng 585true P<0.05), and the expressions of TLR4, HMGB1 mRNA and protein were significantly increased (P<0.05); the levels of TNF-α and IL-1β in the spinal cord were also significantly increased after CCI compared with those in the sham goup (P<0.05). Curcumin markedly attenuated CCI- induced mechanical allodynia and thermal hyperalgesia through inhibiting the activation of TLR4 pathway and production of inflammatory cytokines. These results demonstrated that curcumin alleviated neuropathic pain may be by decreasing the expression inflammatory cytokines produced by TLR4 pathway.]]> 2012/2/17 0:00:00 GU Hong-Feng,LIAO Duan-Fang,TANG Xiao-Qing and KUANG Xin 584true 2O2 of L6 rat myoblast cells was investigated in this study. The hydrogen peroxide(H2O2) was used to establish the H2O2-induced apoptosis model by L6 rat myoblast cells that were treated with 0.3, 0.15, 0.75 g/L oxymatrine. The survival rate was measured by MTT method，cell cycles and apoptosis rate of L6 rat myoblast cells were detected by flow cytometry, the injury degree was determined by apoptosis cytochemistry fluorescent antibody of Bcl-2 and Bax, DAPI staining and HE staining. The protein differences were examined by Western blot. Results showed that survival rate of L6 myoblast cells was degraded and the apoptosis rate was increased by damaging H2O2. The survival rate of L6 myoblast cells was heightened by oxymatrine of each dose. They could induce the increase in expression of Bcl-2 and the decrease in expression of Bax. Its degree of protection was increased with raising the dosage of oxymatrine and the protective effect of 0.3 g/L dosage is obvious. The protective effect of 0.15, 0.75 g/L oxymatrine were secondary. The main physiological and biochemical effects of oxymatrine was related to protect rat myoblast apoptosis model through NFκB signal transduction. These results implied that oxymatrine may have the potential as a new antioxidant treatment medicine.]]> 2012/3/23 0:00:00 ZHU Dao-Li,WANG Kang-Le,CHEN Pei-Lin and JIANG Ya-Qiong 583true INK4a and sonic hedgehog signal pathway in cervical squamous cell carcinoma and its precursor lesions. The expression of P16INK4a, Smo, Ptch and Gli in different HPV types positive cell lines were detected by Western-blot. A tissue microarray constructed with 20 normal cervical tissues and 100 uterine cervical cancers and related lesions (28 squamous cell carcinomas, 26 cervical intraepithelial neoplasia (CIN)Ⅲ, 16 CINⅡ, 12 CINⅠ, 18 tumor-adjacent tissue specimens) was immunohistochemically analyzed with anti- P16INK4a, Shh, Patched (Ptch), Smoothened (Smo), Gli antibodies. The correlation between their expressions was analyzed. There was no significant difference among different HPV type cell lines regarding the expression of P16INK4a and Shh, Ptch and Gli proteins(P>0.05). The expression of P16INK4a and the Hh-signaling molecules was greatly enhanced in cervical carcinoma tissues, compared with that in normal epithelium and tumor-adjacent tissues (P<0.05). There was no significant difference between CINⅠ and normal epithelium(P>0.05), whereas, in case of P16INK4a, Shh, Smo, and Gli, the differences among CINⅠ, CINⅡ and CINⅢ were significant (P<0.05). The expression of P16INK4a protein was significantly correlated with that of Shh, Smo and Gli protein in CINⅡ-CINⅢ and cervical carcinoma and was correlated with that of Shh, Smo only in carcinoma tissue. P16INK4a and the Hh-Gli signaling pathways were extensively activated in the development and evolution of cervical cancer, and the overexpression of P16INK4a was correlated with Hh-signaling pathways. The abnormal Hh-signaling pathways maybe much associated with Smo protein overexpression induced by Shh, which can upregulate the expression of Gli protein.]]> 2012/4/28 0:00:00 MIAO Jin-Wei,ZHANG Yong-Qing,XU Chun-Yu,FANG Chun and DENG Xiao-Hong 582true NBS1 plays a key role in the repair of DNA double strand breaks and the maintenance of genomic stability. It has been shown in recent studies some common NBS1 variants maybe associated with genetic susceptibility of tumors. In the present study, rare allele frequency of single nucleotide polymorphisms (SNPs) of NBS1 gene in primary liver cancer were detected by the method of high resolution Single Strand Conformation Polymorphism (SSCP) analysis, with the aims to analyze the correlation between NBS1 SNPs and primary liver cancer, and evaluate the applicability of the high resolution SSCP technique in the genotyping of SNPs. The rare allele frequency of NBS1 SNPs were detected in 327 cases of primary liver cancer and 295 negative controls from Han people of China by the method of high resolution SSCP analysis. The correlation was analyzed between NBS1 SNPs and primary liver cancer. Genotyping of six common NBS1 SNPs in part samples was carried out by both SSCP analysis and direct sequencing simultaneously to compare the difference between them and evaluate the accuracy and applicability of SSCP analysis in genotyping of SNPs. The results from the tissue samples showed, among six NBS1 SNPs(102G>A, 320+208G/A, 553G>C, 1197T>C, 2016A>G and 2071-30A>T), the rare allele frequency of NBS1 SNP 1197T>C was significantly higher in the 119 cases of primary liver cancer (68.1%) than that in the 95 controls of cirrhosis/chronic hepatitis B (57.9%)(P=0.0298). Similar results was obtained by detection with blood samples of 208 cases of hepatocellular carcinoma(66.8%) and 200 cases of health controls(58.8%)(P=0.0170). There was no any significant difference of the rare allele frequency of the other five NBS1 SNPs. The same result was observed for genotyping of NBS1 SNPs either by the high resolution SSCP analysis or by the direct sequencing. However, higher quality of PCR products is requested for the direct sequencing compared with SSCP analysis. These results suggest that the NBS1 SNP 1197T>C may be associated with the risk of primary liver cancer in the Chinese Han population. High resolution SSCP analysis, with the same accuracy as direct sequencing in the genotyping of NBS1 SNPs and easier to handle, is suitable for the genotyping of multiple known SNPs in large scale cohort study.]]> 2012/4/5 0:00:00 HUANG Jian,ZHAO Yan-Ping,LI Qian,ZHANG Jun-Xia,WANG Yan and ZHANG Bei 581true C. elegans genome encodes only one Rim gene, unc-10. In this study, we demonstrate that UNC-10 is involved in the dense core vesicle (DCV) exocytosis in C. elegans neurons. We find that unc-10 mutants exhibit reduced peptide release in vivo. In IDA-1::GFP labelled ALA neurons, which serve as a good model for studying DCV exocytosis, mutations of UNC-10 lead to a visible reduction of the readily released pool (RRP) size. The analysis of vesicular docking by total internal reflection fluorescence microscopy (TIRFM) reveals that the loss of function of unc-10 hinders the docking of DCVs in neurons. The above results provide evidence that UNC-10 is required for the exocytosis of DCVs and may be involved in the docking step of DCV release.]]> 2011/9/21 0:00:00 FENG Wan-Juan,ZHOU Wei,ZHANG Yong-Deng,WU Zheng-Xing and XU Tao 580true +-dependent deacetylase, reduces apoptosis in several different cells. However, the role of Sirtuin 1 in apoptosis of disc nucleus pulposus (NP) cells remains unclear. The present study was performed to determine whether degenerative human NP would express Sirtuin 1, and to investigate the role of Sirtuin 1 in NP cells apoptosis. Here we show that Sirtuin 1 mRNA and protein levels in disc NP from patients (< 25 years) with lumbar vertebra fracture (LVF) are greater than those in degenerative disc NP from patients (> 55 years) with lumbar disc herniation (LDH). The rate of apoptosis was far fewer in resveratrol-treated NP cells than that in siRNA transfected or nicotinamide-treated NP cells. We also tried to explore the signaling molecules that mediate the protective property of Sirtuin 1 by Western blot and inhibitor analysis. After Sirtuin 1 siRNA transfected, NP cells decreased phosphorylation of Akt, while resveratrol phosphorylated Akt. Treatment with LY294002 or Akt siRNA increased the rate of apoptosis. Our results indicate that Sirtuin 1 plays a critical role in survival of degenerative human NP cells through the Akt anti-apoptotic signaling pathway.]]> 2012/2/3 0:00:00 WANG Da-Wu,HU Zhen-Ming,HAO Jie,HE Bin,GAN Qiang,ZHONG Xiao-Ming,SHEN Jie-Liang and FANG Ji 579true in vitro (DIV5) to display the morphological details of the dendrites and dendritic protrusions. Different doses of rosiglitazone (5, 10 and 20 μmol/L) or 20 μmol/L rosiglitazone with the presence of 5 μmol/L GW9662, an antagonist of PPARγ, were applied to neurons for 24 h at DIV6. Live-cell imaging technology was used to investigate the effects of rosiglitazone on the development of dendritic filopodia and dendritic tree in hippocampal neurons at DIV7. Our results have shown that rosiglitazone increased the density of dendritic filopodia in a dose-dependent manner. The filopodia density of cultured neurons in 10 μmol/L ((34.27 ± 2.12)/100 μm, n = 21) and 20 μmol/L ((37.75 ± 2.09)/100 μm, n=21) rosiglitazone treated group was significantly increased compared with those of control group((26.45±1.47)/100 μm, n=21), but the filopodia density of cultured neurons in 5 μmol/L rosiglitazone treated group ((27.66 ± 1.84)/100 μm, n=20) was not significantly altered compared with those of control group. Rosiglitazone treatment of 5, 10 and 20 μmol/L concentrations has no effect on the length and motility of dendritic filopodia. In addition, neither the length nor the number of dendritic branches was altered by treatment of 5, 10 and 20 μmol/L rosiglitazone. 5 μmol/L GW9662 attenuated the increased filopodia density induced by 20 μmol/L rosiglitazone. These results suggest that rosiglitazone may affect the initial stage of hippocampal neuron development through the PPARγ pathway. This may be a possible mechanism of the neuroprotective effects of rosiglitazone.]]> 2012/3/13 0:00:00 LIU Gui-Lan,BAO Xiao-Ming,LU Zhou-Yi,XU Shu-Jun and WANG Qin-Wen 578true 2011/10/14 0:00:00 WANG Zhong-Feng,WANG Xin-Lu and GAO Guang-Xia 577true Oryza sativa), of which OsWAK50 is a typical RLK with an extracellular domain, a transmembrane domain and an intracellular kinase domain. OsWAK50-GFP fusion protein is localized on the cell surface and associated with the cell wall. By the yeast two-hybrid screen, we identified 20 possible OsWAK50 intracellular interacting proteins. Further one to one reverse hybridization indicates that OsSK4, OsSWIB and OsSWI3C interact with OsWAK50 intracellular domain. OsSWIB interacts directly with the kinase domain of OsWAK50, while OsSK4 and OsSWI3C interact with OsWAK50 intracellular domain in a C terminal dependent manner. OsSK4 and OsSWIB can also interact with OsWAK53a, the closest homolog of OsWAK50 in rice, whereas OsSWI3C can not interact with OsWAK53a. The interaction between OsWAK50 and OsSK4, OsWAK53a and OsSK4 was further confirmed by in vivo BiFC assay. These results provide important clues for elucidating the molecular mechanisms of OsWAK50 function in rice.]]> 2011/12/5 0:00:00 SUN Li-Jing,ZHANG Qian,LU Tie-Gang and SUN Ying 576true in vivo treatment effect of CD (cytosine deaminase) suicidal genetic system on the glioma in rat was explored. The lentivirus carrier which was composed of CD genes was fabricated and the mouse BMMSCs were transfected by such CD genes to obtain constantly expressed cells, then BMMSCs were transplanted into the animal model which was built through an intracranial stereotactic inoculating method using a group of 40 SD rats. The rats were divided into 5 groups uniformly according to the cell type inoculated, i.e. ① C6 glioma, ②C6+MSCs(mesenchymal stems cells) cells (1∶1), ③C6+MSC cells (1∶2), ④C6+MSC -codA/eGFP cells (1∶1) and ⑤C6+MSC-codA/eGFP cells (1∶2). After 7 days of tumor formation, the abdominal cavity of rats were injected with 5-FC at 500 mg/(kg·d) for 14 days. Intensive scanning was carried out weekly to observe tumor volume using MRI. Simultaneously, survival times, routine pathological test, RT-PCR assay and HE staining were also operated. The results of MRI showed that the focus of infection in group ① presented round shape. The necrosis area of tumor found in the center, achieved an average volume of 246 mm3, and the survival time of the center area was 15.3 days. About the group ② and ③, the survival times were 16.0 and 16.6 days respectively. But in group ④ and ⑤, the survival times were both larger than 30 days, the necrosis area of tumor was 55 and 40 mm3 respectively at day 14, and the inhibition efficiency was 77.24% and 83.28% respectively after 28 days of treatment. It was concluded that the MRI scanning could clearly show the volume, shape, and internal structure of the tumor, which was highly related to the results of pathology. The treatment effect of bone marrow MSCs with CD genes and 5-FC therapy system on C6 intracranial glioma could be verified through dynamic MRI observation. Moreover, the RT-PCR test confirmed the expression of cytosine deaminase inside tumor organization.]]> 2011/12/20 0:00:00 SONG Fei,XING Qi,SONG Ke-Dong,LIU Jian,JI Guang-Chun,MA Yu-Fang,LIU Tian-Qing,MA Xue-Hu and TIAN Xiao-Feng 575true Ik after exposure to EMF, and with the increase of the voltage depolarization, the inhibition rates of 1 mT and 5 mT almost unchanged and the inhibition rates were (30.0 ± 4.2)% and (20.0 ± 2.2)%，respectively. While the inhibition rate of 10 mT became larger and the maximum inhibition rate was 43.4%. Additionally, 1 mT and 5 mT magnetic fields both affected the activation characteristics of delayed rectifier potassium channel, the half activation voltage became larger and the slope factor unchanged, while 10 mT magnetic fields did not changed anything. This paper indicated that the structure and function of the channel protein on cell membrane may be altered by 50 Hz EMF, and there were different effects on the channel for different strength of magnetic fields, the window effects of strength of magnetic fields were improved in this study.]]> 2011/12/5 0:00:00 LI Gang,LI Dan-Dan,LI Yuan-Yuan and LIN Ling 574true fabI1 and fabI2 (UniProt AC: Q57A95 and Q57EU5), annotated as encodes putative enoyl-ACP reductase in Brucella abortus genome. Sequence alignment found that BaFabI1 and BaFabI2 are 50% and 51% identical to E. coli FabI, respectively. Further analysis identified that the catalytically active triad (Tyr-(Xaa)6-Lys) of E. coli FabI are present in both BaFabI1 and BaFabI2. Expression of either of the two proteins restores the growth and the fatty acid synthesis of the E. coli fabI temperature sensitive mutant JP1111 under nonpermissive condition. In vitro assay identifies that both proteins restore the fatty acid synthetic ability and are active with substrates of all fatty acid chain lengths. These results demonstrated that B. abortus possesses two FabI-like enoyl-ACP reductases and it represents a new kind of diversity of bacterial enoyl-ACP reductase.]]> 2011/12/20 0:00:00 LEI Ming,MA Jin-Cheng and Wang Hai-Hong 573true 70 000 structures), it is actually of high possibility to find a structural homologue for a newly identified sequence. In this work, we present a new method that predicts β-turns by combining homology information extracted from PDB with the results predicted by NetTurnP. Two datasets, the golden set BT426 and the self-constructed dataset EVA937, are used to assess our method. For each sequence in both datasets, only homologues deposited earlier than the sequence in PDB are employed. We have achieved Matthews correlation coefficients (MCCs) of 0.56, 0.52 respectively, which are higher than those obtained by NetTurnP alone of 0.50, 0.46, and the prediction accuracies (Qtotal) obtained using our method are 81.4% and 80.4% separately, while NetTurnP alone achieves 78.2% and 77.3%. The results confirm that combining the homology information with state-of-the-art β-turn predictors like NetTurnP can significantly improve the prediction accuracy. A Java program called BTMapping has been written to implement our method, which is freely available at http://www.bio530.weebly.com together with the related datasets.]]> 2011/11/14 0:00:00 QIAN Gang,WANG Hai-Yan and YUAN Zhe-Ming 572true Cecropin B gene was modified according to the codon bias of the nuclear genome in Chlamydomonas reinhardtii. Four repeats of the Cecropin B gene were fused in tandem and each repeat was separated by inserting a cleavable linker peptide sequence (LWMRFA). The artificial DNA (522 bp in length) was inserted into a site between hsp70-RBCS2 promoter and RBCS2 terminator for constructing the expression vector pCB124. A cell-wall deficient strain of C. reinhardtii CC-849 was transformed by using glass bead method with pCB124. A large number of transformants were selected on Tris-acetate-phosphate media containing 10 mg/L Zeomycin. PCR and RT-PCR analyses on the transformants revealed that tandem repeated Cecropin B gene had been integrated into the genome of C. reinhardtii and could express at transcriptional level. The Western blot results confirmed the presence of recombinant antimicrobial peptide Cecropin B in the transgenic algal cells. The total protein was extracted from transgenic algae and its antimicrobial activity was tested. The results indicated that the extracted proteins from transgenic alga showed very strong antimicrobial activity against both Gram negative bacterium (E. coli JM109) and Gram positive bacteria (Bacillus subtilis and Micrococcus lysodeikticus). This finding has provided a new approach for production and utilization of antibacterial bait-algae.]]> 2011/10/14 0:00:00 MU Fei-Yun,LI Hui and HU Zhang-Li 571true Ampelopsis grossedentata is known to have an anti-proliferative effect in vitro on a few cancer cells. However, the exact mechanisms retain unclear. The aim of this article was to study the effect of DMY on cell invasion of highly metastatic human breast cancer MDA-MB-231 cells, and investigate the possible mechanisms. The anti-proliferation effect of DMY on MDA-MB-231 cells was determined by the MTT assay. The gelatinolytic activity was assessed by gelatin zymography. The mRNA and the protein expression levels of MMP-2/-9 were evaluated by Real-time PCR and Western blot assay, respectively. The effect of DMY on cell invasion was observed by a transwell model. Results indicated that DMY inhibited the proliferation of MDA-MB-231 cells in a dose-dependent manner, and the IC50 value was about 73.6 mg/L after treated by DMY for 48 h. DMY suppressesed gelatinase activity and MMP-2/-9 protein expression levels in a dose-dependent manner, and restrained MMP-2/-9 mRNA expression levels. In addition, DMY inhibited the invasion of MDA-MB-231 cells in a dose-dependent manner without cytotoxicity against the cancer cells. DMY can inhibit proliferation and invasion of MDA-MB-231 cells, and invasion inhibition may be related to the down-regulation of the expression levels of MMP-2/-9 proteins.]]> 2011/11/29 0:00:00 ZHOU Fang-Zhen,ZHANG Xiao-Yuan,ZHAN Yuan-Jing and GUO Yong 570true 2011/7/29 0:00:00 ZHAO Qian,XIAO Jing-Fa and YU Jun 569true CIQ) might be mediated by recombination rate. In this study, a parameter is defined to characterize GRCIQ, and the correlation between GRCIQ and recombination rate is analyzed in the human genome. The results show that there is a positive correlation between them, indicating recombination is likely to be an important pathway to increase GRCIQ.]]> 2012/2/17 0:00:00 LIU Guo-Qing and LUO Liao-Fu 568true trans retinoic acid (atRA) (0.2, 0.67, 2.0 and 6.7 μmol/L) on cell proliferation and apoptosis, and then examined the role of gamma amino butyric acid (GABA) signaling pathway in regulation of cell proliferation and apoptosis by atRA in murine embryonic palate mesenchymal (MEPM) cells. Results showed that atRA (2 μmol/L and 6.7 μmol/L) significantly inhibited cell proliferation and increased apoptosis. The mRNA and protein expression of glutamic acid decarboxylase 67 (GAD 67) which was a key enzyme in synthesis of GABA were significantly down regulated by atRA (0.67, 2.0 μmol/L and 6.7 μmol/L). But the mRNA and protein expression of GABAAR-β3 were shown no obvious change compared with the control group. When GABA (1.0 μmol/L) was added to cell culture system, the effect of atRA (6.7 μmol/L) on the proliferation and apoptosis of MEPM cells was reversed. In conclusion, all-trans retinoic acid inhibits cell proliferation and promotes cell apoptosis through gamma amino butyric acid pathway in murine embryonic palate mesenchymal cells.]]> 2011/6/30 0:00:00 WEI Xue-Tao,QIAO Yang-Zheng,SHANG Lan-Qin and LI Yong 567true Pantoea agglomerans YS19 is an endophytic diazotrophic bacterium isolated from rice (Oryza sativa cv. Yuefu) grown in temperate climatic regions in west Beijing (China). The bacterium forms aggregate structures called "symplasmata", in which several (at least two) to hundreds of individual cells tightly bind together. Our previous study revealed that there were two growth stages for YS19, including the single cell stage existing before exponential growth phase and the symplasmata forming stage starting at the end of the exponential growth phase in liquid LB medium. More strikingly, the symplasmata structures contribute to bacterial stress (e.g., dehydration, heavy metal toxicity, and osmotic shock) resistance and are especially significant for bacterial surviving strategy in suiting an adaptive life to hostile environments. In this research, YS19 was cultivated in LB medium, and the whole cellular protein expression of the cultures sampled at different times (0～12 h) was analyzed by SDS-PAGE. Here, a novel protein differentially expressed at the symplasmata forming stage was captured. The protein was purified and digested by trypsin. The digests were analyzed by MALDI-TOF mass spectrometry and the peptide mass fingerprint of this protein was successfully obtained. Then, database searching with Mascot in the SWISS-Prot database was performed using the recorded peptide mass fingerprint data. It is found that among 53 peptides, there are 14 peptide masses matching to MalE from Enterobacter aerogenes, which is a protein belonging to the periplasmic maltose-binding protein family of the ATP-binding cassette transporters. Then the gene of the MalE protein was cloned from YS19, expressed in the E. coli BL21(DE3), and the recombinant protein was purified to homogeneity. Under stress (acid) treatments, the recombinant protein showed strong anti-aggregation ability, and even exhibited chaperone-like activity. This assay was achieved by comparing the quantity of the substrate proteins remaining in the supernatant under various denaturing acid conditions, where the aggregation of the substrate protein was effectively suppressed in the presence of the recombinant MalE protein. Far-UV circular dichroism spectroscopy examination on the purified MalE proteins under different pHs indicated that the global structures exhibited a sudden transition from a highly ordered conformation to a less ordered one. However, MalE was still capable of maintaining most of its secondary structures under these extreme acidic conditions (pH1～2).Finally, the Bis-ANS fluorescent probe technique was used to explore the effect of acid condition on the hydrophobic surfaces exposing of the protein. It is found that with the decrease of the pH value, the fluorescence intensity of the samples increased gradually, suggesting the MalE protein exposed the hydrophobic surfaces of the molecule to bind the substrates, implying the protein performed its chaperone activity via exposing its hydrophobic surfaces under acidic condition. MalE protein, originally found as a periplasmic maltose-binding protein of the ATP-binding cassette transporters, whose expression in YS19, however, far exceeds the amount being needed for transmembrane transports, is most likely a moonlighting protein for its alternative biological functions as a chaperone protein as revealed by this study. This research provided valuable information on the life styles and survival strategies of microorganisms that forms multicellular aggregates at specific growth stages and also explained the protein basis adopted by YS19 symplasmata-forming cells in surviving the stress conditions.]]> 2011/12/20 0:00:00 LI Qian-Qian,XIONG Xiao-Lu,MU Yu-Fei,YI Ting,LI Er-Li,LU Zhen-Yue,MIAO Yu-Xuan and FENG Yong-Jun 566true 2011/11/14 0:00:00 WANG Duan-Qing,HE Tao,WANG Li,WANG Yu-Min and SHAO Wei-Dong 565true 6±2.54×104) vs wild type (1.17×108±1.46×107), P < 0.001). The infectivity of virus released into the supernatant significantly reduced similarly. Further study found that these five mutations changed secondary structure of HCV core gene. In order to further explore the underlying reasons for the weakening of J6JFH1/ΔF virus replication and infection, we constructed five separate mutant viruses based on J6JFH1, respectively, and form J6JFH1m1 nt406T-A, J6JFH1m2 nt433T-A, J6JFH1m3 nt472G-A, J6JFH1m4 nt479/481G-A, G-A, J6JFH1m5 nt613C-A. These 5 virus mutants do not influence the secondary structure of the core region, while J6JFH1m1, J6JFH1m2, J6JFH1m3, J6JFH1m4, J6JFH1m5 respectively stopped F protein expression shifting from nt374-383, nt417-419, nt436-445, nt474-477, nt597-605, in which J6JFH1m5 stopped all reported forms of F protein. By in vitro transcription and transfection into Huh7.5.1 cells, 48 hours post-transfection, the detection of intracellular viral protein NS5A expression levels, viral RNA in transfected cells, and virus titers in supernatant were carried out. The results showed that no significant differences were found among groups five separate mutants and wild type. These results indicated that the lack of F protein itself did not affect HCV viral replication and virion assembly and infectivity. All the results were seemed to result from the secondary structure of the core gene, but the mechanism was yet to be further studied. We concluded that HCV F protein deficiency did not affect translation of viral replication and packaging and release of virus particles, the core gene secondary structure changes had a great impact on viral replication and protein expression.]]> 2011/7/26 0:00:00 WANG Wen-Bo,XU Gang,WANG Yan,TAO Qing-Yuan,REN Hao and QI Zhong-Tian 564true IC50 of adriamycin to thermotolerant HepG2/TT cells and enhanced adriamycin-induced apoptosis in HepG2/TT cells in a concentration-dependent manner. The levels of P-gp and Hsp70 in HepG2/TT cells were obviously higher than that in HepG2 cells. Treatment with curcumin (10 μmol/L) for 24 h significantly reduced the levels of P-gp and Hsp70 in HepG2/TT cells. The accumulation of intracellular adriamycin in HepG2/TT cells was markedly lower than that in HepG2 cells and curcumin (10 μmol/L for 3 h) significantly increased the level of intracellular adriamycin accumulation in HepG2/TT cells. HepG2/TT cells obviously inhibited caspase-3 activation triggered by adriamycin, but co-treatment with 10 μmol/L of curcumin for 24 h significantly augmented the activation of caspase-3 in HepG2/TT cells treated with adriamycin. Curcumin overcomes the adriamycin-resistance of thermotolerant HepG2/TT cells by promoting adriamycin-triggered caspase-3 activation through down-regulating the activity and expression of P-gp and the expression of Hsp70.]]> 2011/7/29 0:00:00 QIN Chun-Hong,LI Yong-Guo,WU Jiang and HE Hong-Jie 563true et al. The results indicated that Embelin remarkably inhibited human hepatocarcinoma cell Bel-7404 growth. Further experiments found that the mechanism of tumor cell growth inhibition by Embelin was due to the cell apoptosis with the activation of caspase apoptotic pathway and blockage of NF-κB signal pathway. The studies laid a base on the further treatment of human hepatoma carcinoma by using Embelin.]]> 2011/7/26 0:00:00 WANG Shi-Bing,LEI Wen,XU Yan-Hua,GUO Ke-Ni and WANG Yi-Gang 562true -/- mice, 8 weeks old, randomly divided into three groups: ①normal food group, ② high fat group, ③AMD3100 group. Animals from high fat group and AMD3100 group were fed with western food which including 15% fat and 0.25% cholesterol. After feeding 12 weeks, all mice were anesthetized by 0.2～0.3 ml Urethane (20%) and removed eyeball in order to obtain blood preparation. Serum lever of SDF-1α was measured by ELISA. Serum triglyceride (TG), total cholesterol (TC), and high density lipoprotein cholesterol (HDL-C) were determined by commercially enzymatic methods. The Hematoxylin/Eosin dyeing of paraffin section was used to detect the area of atherosclerotic plaque of apoE-/- mice aortic root transaction. The expression of CXCR4, TNF-α and NF-κB was analyzed by real time RT-PCR and Western blot, respectively. As a result, AMD3100 treatment resulted in a significant increase of atherosclerotic lesion area and the expression of TNF-α and NF-κB in apoE-/- mice. Serum TG, TC, HDL-C and LDL-C concentrations were not markedly changed. AMD3100 reduced SDF-1α serum lever and CXCR4 expression on artery wall. It can be concluded that, atherosclerotic lesions in apoE-/- mice were aggravated by long term administration of CXCR4 antagonist AMD3100. Possible mechanisms of this action for AMD3100 are associated with the up-regulation pro-inflammatory factors TNF-α and NF-κB and down-regulation SDF-1α/CXCR4 axis.]]> 2011/9/30 0:00:00 WANG Zuo,SU Wei,ZHOU Xiao-Feng,ZHANG Kai,LI Shuang,MA Xiao-Feng and JIANG Zhi-Sheng 561true Caenorhabditis elegans FAT-1 and FAT-2 genes, which encode ω-3 and Δ-12 fatty acid desaturases respectively and human catalase hCAT gene, based on "self-cleaving" 2A peptides, and they were generated by microinjection. Fatty acids in skeletal muscle were analyzed by gas chromatography. The level of total n-3 polyunsaturated fatty acids in transgenic mice was 2.6-fold higher than that in wild type mice, while there was no significantly difference in the level of total n-6 polyunsaturated fatty acids between transgenic and wild type mice. But the skeletal muscle tissue of transgenic mice had a dramatically reduced ratio of n-6/n-3 fatty acids (P < 0.01). In addition, the expression level of human catalase was identified by Western blot, and the catalase activity of hCAT in skeletal muscle of transgenic mice was significantly higher than that of wild type mice (P < 0.01).]]> 2011/9/21 0:00:00 FANG Rui,PENG Yun-Qian,ZHENG Min and MENG Qing-Yong 560true BDD-FⅧ gene. Here, we plan to achieve this by making Cys mutations at the Met662 in A2-domain of heavy chain and Asp1828 in A3-domain of light chain. By co-transfecting cultured COS-7 cells transiently with mutant heavy and light chain genes, a disulfide cross-linked heavy and light chain was observed by Western blotting. The secretion of heavy chain and coagulation activity were up to (109±13) μg/L and (0.65±0.12) U/ml respectively analyzed by ELISA and Coatest method greater than that of two chains of wild-type BDD-FⅧ gene co-transfected cells((71±11) μg/L and (0.35±0.05) U/ml. It suggests that inter-chain disulfide linking could efficiently improve efficacy of dual-vector delivery of the BDD-FⅧ gene. It provided evidence for ongoing in vivo BDD-FⅧ gene delivery by dual-AAV vector.]]> 2011/11/23 0:00:00 ZHU Fu-Xiang,YANG Shu-De,LIU Ze-Long,MIAO Jing,QU Hui-Ge and CHI Xiao-Yan 559true 111Cd concentration to indicate the concentration of QDs in plasma and organs. The fluorescence images show that QDs can penetrate into the dermal layer and deposit in the organs through blood circulation. ICP-MS result indicates that QDs deposit seriously in liver and kidney, and they are difficult to clear. 111Cd concentration is still more than 14 ng/g in kidney after 5 days. These results suggest that QDs has in vivo transdermal delivery capacity through mouse skin and is harmful to the liver and kidney.]]> 2011/9/30 0:00:00 TANG Lei,ZHANG Chun-Ling,SONG Guang-Ming,XU Zhong-Wei and JIN Xun 558true 2+ as cofactors and a smaller regulatory subunit bound to the end-product feedback signals such as valine, leucine or isoleucine to down regulate the holoenzyme activities. The Escherichia coli ilvN gene that encodes the regulatory subunit of AHAS Ⅰ was cloned into pET28a and expressed in soluble form at high levels in E. coli strain BL21 (DE3). The protein was purified using Ni2+-chelating chromatography followed by size-exclusion chromatography. Crystals of IlvN protein were obtained and diffracted to 2.6 Å. To further study the regulatory mechanism, crystals of IlvN co-crystallized with its effector valine were also obtained and diffracted to 3.0 Å.]]> 2011/6/16 0:00:00 NIU Xu-Hui,LIU Xiang,ZHOU Yan-Fei,XI Zhen and SU Xiao-Dong 557true ARF2, we isolated the SlARF2 gene and analyzed its molecular features, in addition, we observed the subcellular localization of ARF2 in transgenic tomato plants. Physicochemical properties and molecular features of ARF2 were predicted by bioinformatic approaches including physical and chemical properties analysis, hydrophobicity analysis, domain analysis, phylogenetic tree analysis and subcellular localization analysis. Moreover, the full-length of SlARF2 gene was amplified by RT-PCR, and a binary vector consisting of ARF2 fused with the yellow fluorescent protein (YFP) coding sequence was further constructed. Using the method of Agrobacterium-mediated transformation, the recombinant vector was transformed into wild-type tomato, and the transgenic tomato plants were confirmed by PCR. The roots of transgenic tomato seedlings were observed under a fluorescence microscope. Bioinformatic analysis showed that the SlARF2 is an unstable soluble protein, which is rich in Ser, Gly, Pro and Leu residues, and it has typical structure and functional domain of ARF family. Tomato ARF2 shares 70.08 %, 66.94 %, and 60.87 % similarity in amino acid sequences with grapes, cassava and Arabidopsis, respectively. Analyses of restriction enzyme digestion and sequencing confirmed the expression vector pBA-ARF2-YFP was successfully constructed. PCR results showed the fusion protein was expressed in transgenic tomato plants. The subcellular localization of SlARF2 was found mostly in the nucleus. SlARF2 has the key characteristics of transcription factor, which is localized in the nucleus, and it might play a pivotal role in tomato fruit development and ripening. This might provide references for studying the function of SlARF2 gene.]]> 2011/8/24 0:00:00 FENG Yuan-Yuan,HOU Pei,LI Ying-Nan and LIU Yong-Sheng 556true 2011/6/30 0:00:00 ZHAO Lei,SANG Chen,YANG Chun and ZHUANG Feng-Yuan 555true 2011/8/24 0:00:00 GU Huan,YAN Lu,LI Jia and ZHANG Gui-Ying 554true 2011/8/24 0:00:00 LIN Bei-Bei,ZHANG Yi,XU Yang,PEI Chen-Lin,CAO E-Xiang,PENG Guo-Qing,ZHU Xin,ZHAO Yan-Hua and CAI Jing-Ting 553true 2013/2/28 0:00:00 LI Xiao-Jian,JIANG Zhen and WANG Yi 552true in vitro and in vivo assays were conducted to evidence whether BMP9 can regulate osteogenic differentiation of mesenchymal stem cells through ERK1/2 kinase pathway. The results showed that BMP9 can activate ERK1/2 kinase through increase the phosphorylated form of ERK1/2 kinase. ERK1/2 kinase inhibitor PD98059 can increase the ALP activity, OPN expression and calcium deposition of C3H10T1/2 cells induced by BMP9. Furthermore, PD98059 also led to enhancement of Runx2 activity and activation of canonical Smad pathway stimulated by BMP9. When ERK1/2 kinase was silenced by RNA interference, BMP9-activated Smad pathway was further enhanced. Moreover, ALP activity, calcium deposition and in invo ectopic bone formation were accordingly increased along with knockdown of ERK1/2 kinase. Taken together, those results intensively suggested that BMP9 can activate ERK1/2 kinase, and inhibition of ERK1/2 kinase can enhance BMP9-induced osteogenic differentiation of mesenchymal stem cells. ERK1/2 are highly capable of negatively regulate BMP9-induced osteoblastic differentiation of mesenchymal stem cells.]]> 2013/2/28 0:00:00 SONG Tao,HE Juan-Wen,WANG Jin,TANG Min and LUO Jin-Yong 551true In vitro attachment and outgrowth assays were performed in EPCs, and transwell chambeer was performed in decidual cells. In vivo, mouse from D8 to D13 were treated by intraperitoneal injection of different concentrations of GABA . On D14, the placenta structure was examined with the histological method. The results suggested that GB1 mRNA and protein dynamic expressed from D1 to D8 uterus. 100 μmol/L GABA and 5 μmol/L baclofen promoted EPCs outgrowth and surppressed decidual cells invasion. At the same time, 20 μmol/L 2-hydroxysaclofen could reversed both of the functions of GABA on EPCs and decidual cells. The histological structures of placentas changed appearantly on D14. In the layer of labrinthine, the cells arranged crowdly and maternal blood and fetal rat vascular decreased number or dysplasia. In spongiotrophoblast, cytotrophoblast cells becomed smaller and glycogen cells decreased or disappeared. The combined results indicated during the early-middle mouse placenta establishment GABA signal was in favour of trophoblasts invasion but played converse role in decidual cells and damaged placenta structure.]]> 2013/2/28 0:00:00 LUO Wen-Ping,TAN Dong-Mei,YANG Gen-Ling,LU Jun-Jie,ZHAO Hai and TAN Yi 550true Helicobacter pylori is identified as a new carcinogenic factor. Tipα induces high expression of TNF-α through NF-κB activation, thus promoting host inflammation and tumor progression. The homodimer of Tipα as its active form for functional performances is cross-linked by a pair of inter-molecular disulfide bridges (Cys25-Cys25 and Cys27 and Cys27). Tipα (25～192) was cloned into pET22b and expressed as soluble protein in E. coli strain BL21 (DE3). Recombinant active Tipα was first purified through Ni2+-chelating chromatography, and then further purified by cation-exchange chromatography and size-exclusion chromatography to get the pure homodimer protein. Native Tipα and SeMet Tipα were crystallized and optimized using hanging drop and microbatch methods, with diffraction to 2.2? and 2.6?, respectively. These crystals belonged to C2 space group with similar unit-cell parameters. The native protein crystal had unit-cell parameters a=127.01?, b=47.57?, c=96.5?, α=γ=90°, β=127.5°. An attempt to solve the three-dimensional structure of this protein by MAD method is under way.]]> 2013/2/28 0:00:00 GAO Ming-Ming,ZHANG Ying and WANG Da-Cheng 549true in vitro through improving protein trans-splicing. In this study, a pair of plasmid vectors expressing human BDD-FⅧ heavy and light chain fused with lucine zipper and split Ssp DnaB intein was co-injected into C57BL/6 mice via the portal vein. Fourty-eight hours post-injection, the level of heavy chain and FⅧ coagulation activity in collected plasma were determined and shown as (298±67) μg/L and (1.15±0.29) U/ml respectively, greater than that of control mice injected with both vectors without leucine zippers ((179±59) μg/L and (0.58±0.19) U/ml). It demonstrated that leucine zippers fused intein could increase FⅧ coagulation activity in plasma of mice with intein-based dual-vector BDD-FⅧ gene delivery through improved protein trans-splicing. It provided evidence for ongoing hemophilia A gene therapy using dual-AAV vecors.]]> 2013/2/28 0:00:00 ZHU Fu-Xiang,LIU Ze-Long,MIAO Jing,QU Hui-Ge and CHI Xiao-Yan 548true Drosophila CAF-1, dCAF-1-p55, together with the other two subunits dCAF-1-p180 and dCAF-1-p105, participates in the deposition of histone H3 and H4 onto the newly synthesized DNA. In addition to the CAF-1 complex, dCAF-1-p55 is also found in several other complexes such as NURF, PRC2 and Sin3-HDAC1 complex, suggesting that it plays multiple roles other than a histone chaperone. Here we report a new allele of dCAF-1-p55 generated by gene targeting. We show that dCAF-1-p55 loss-of-function led to developmental delay and lethality. Further investigations demonstrated that dCAF-1-p55 deficient cells exhibited defects in metaphase chromosome condensation, sister chromatids association and anaphase chromosomes segregation, which are premonitory signs of chromosome instability (CIN). Taken together, these results indicate that dCAF-1-p55 plays a role in the maintenance of chromosomal stability, likely to protect the cells from CIN and the proneness to cancer.]]> 2013/3/25 0:00:00 WU Qing-Hua,LIU Ji-Yong,CHEN Yi-Xu and JIAO Ren-Jie 547true 2013/3/25 0:00:00 PENG Yin-Ting,PU Qing,SUN Xin-De and ZHANG Ji-Ping 546true in vivo. In this study, to improve the stability of FGF21 in vivo, we added two arginines to the C-terminus of FGF21, and named as FGF21-2A. After being calculated by the software pI/Mw, the pI of FGF21-2A reached to 5.84 from 5.43. Then we expressed and purified the mutant protein for the examination of stability and bioactivity. When we used Bandscan5.0 to analyze the supernatant of Rosetta(DE3) lysates after induction, the protein expression levels of FGF21-2A increased by 10.6%. The in vivo half-life time of FGF21-2A was significantly prolonged in pharmacokinetics experiment. And, compared to the wild-type FGF21, FGF21-2A not only had a better capacity of stimulating glucose uptake in 3T3-L1 cells in vitro and stimulating glycogen synthesis of the model mice in vivo, but also provided a significantly long lasting effect on reducing blood glucose in the type Ⅰ and type Ⅱ diabetic animals. Moreover，the results of the real-time PCR showed that, after long term treatment, FGF21-2A could increase the mRNA expression of GLUT1 and GK and reduce the mRNA expression of G6P, which suggests FGF21-2A has the same mechanism of glucose regulation with the wild-type FGF21. In conclusion, modifying FGF21 with arginines increases the protein stability, and enhances the protein bioactivity, which lays the foundation for development of drugs targeting diabetes mellitus.]]> 2013/3/25 0:00:00 HE Kun,ZHANG Ya-Kun,YE Xian-Long,WANG Wen-Fei,CHEN Rui,LIU Ming-Yao,FENG Ming-Fang,XU Jia-Ling and LI De-Shan 545true 2013/3/25 0:00:00 ZOU Zhen-You,WANG Yang,DING Han-Ying,WU Lin-Ping,CHEN Shu-Mei,JIANG Ting,GUO Fang,ZHANG He-Yun,CHEN Xi,ZHANG Qi-Peng,ZENG Ke and ZHANG Chen-Yu 544true in vitro pull-down experiments and immunoprecipitation of complexes assembled in vivo. Subsequently, we showed that the hMBD4 protein expressed in E. coli could be phosphorylated by PrKX. Phosphorylation of hMBD4 by PrKX modulated the binding activity of hMBD4 to fully methylated DNA. These results indicate that MBD4 is a substrate for PrKX.]]> 2013/3/26 0:00:00 LI Wei and LIN Ying 543true 2012/3/1 0:00:00 CHEN Jing-Wen and GUO Liang 542true N and nucleotides were clearly described. We confirmed that the γ-phosphate of ATPγS was not hydrolyzed by Hsp90N. The position of S1 and ATP lid in human Hsp90N-AMPPNP differs significantly from that of the structure of yeast Hsp90-AMPPNP. By analyzing the structure of human Hsp90N-AMPPNP, we found that the interactions of E18-K100 and N40-D127 block the moving of S1 and ATP lid, and then prevent the dimerization of Hsp90N. This reflects the complexity and coordination of Hsp90 on the regulation of the function.]]> 2012/3/6 0:00:00 LI Jian,SUN Li-Hua,XU Chun-Yan,YU Feng,ZHOU Huan,TANG Lin and HE Jian-Hua 541true etc. in eukaryotic system. Previously, Arabidopsis LFR (leaf and flower related) protein localized in nucleus, and its loss-of-function mutants were reported to have pleiotropic phenotypes in leaf, flower and anther development, but the molecular characteristics of LFR protein should be detected further. Firstly, the fusion protein of GAL4 DNA-binding domain (GBD) with full-length of LFR had transactivation activity in yeast Y190 by yeast one-hybrid assay, and at least 2 ARM-repeat domains in the C terminal and full N teminal of LFR were necessary to its transactivation activity. Next, we further analyzed GBD-LFR transactivation activity in Arabidopsis protoplast system, it showed that the transactivation of full-length LFR was not obvious compared with that of a classical transcription activity domain of VP16. In protoplast transient expression system, fluorescence microscopy was used to observed subcellular localization of YFP fusion proteins of truncated or site-mutation LFR, and the data showed that the N terminal 1～25 amino acids of LFR, particular lysine 22 and arginine 4, 23, 25, was responsible for its nuclear localization. In transient co-expressing onion epidermal cell, LFR obviously co-localized with chromatin stucture protein, histone H4, and chromatin-binding protein, HMGA, in nucleus. All these data suggested that LFR might be as one of chromatin-associated proteins to function in Arabidospis development.]]> 2012/3/19 0:00:00 YUAN Can,LI Xiao-Rong,GU Dan-Dan,GU Yue,GAO Ying-Jie and CUI Su-Juan 540true FA), mean diffusivity (MD) and connection strength on fibers linking two brain regions were calculated. Further, these parameters were executed two-sample t-test between CAE group and health control group. Finally, we used Pearson's correlation coefficient to evaluate the relationship between these parameters and epilepsy duration (year). Both CAE and healthy control groups showed similar structural connectivity patterns in DMN. Among these fiber bundles, three were identified in all subjects, with one linking posterior cingulate cortex/precuneus to medial prefrontal cortex, and another two linking posterior cingulate cortex/precuneus to bilateral medial temporal lobes. Furthermore, the significantly decreased FA and connection strength, and increased MD in fiber bundles linking posterior cingulated cortex/precuneus to medial prefrontal cortex, were found in patients compared with the cases in healthy controls (P < 0.05，Bonferroni corrected). Predominantly, the FA values in fiber bundles linking posterior cingulated cortex/precuneus to medial prefrontal cortex were negatively correlated with the epilepsy duration (P=0.006). These findings might reflect the structural basis of the altered functional connectivity in DMN about absence epilepsy. Given that functional connectivity abnormality in our previous work, it is implied that the abnormal fiber connectivity from posterior cingulated cortex/precuneus to medial prefrontal cortex plays an important role in absence epilepsy. This may open up new avenues to understand the pathophysiological mechanisms of absence epilepsy.]]> 2013/9/16 0:00:00 XUE Kai-Qing,LUO Cheng,YANG Tian-Hua,LI Qi-Fu,ZHOU Dong and YAO De-Zhong 539true -/- mice: control group (n=10); IgG group (n=10, 50 μg) and GP Ⅱb/Ⅲa mAb group (n=10, 50 μg). All mice were fed on a Western diet (10% fat and 1.25% cholesterol) for 10 weeks. GP Ⅱb/Ⅲa blockade significantly decreased the atherosclerotic lesion and platelet adhesion to the vessel wall. Immunohistochemistry analysis showed that blocking GP Ⅱb/Ⅲa diminished MOMA-2 and VCAM-1 expression in aortic plaque in ApoE-/- mice. Western blot results indicated that HMGB-1, TLR4, and NF-κB levels were markedly reduced in arteries of ApoE-/- mice treated with GP Ⅱb/Ⅲa mAb (P < 0.05). Moreover, GP Ⅱb/Ⅲa mAb decreased plasma HMGB-1, IL-1β, TNF-α and MCP-1 concentrations. Our findings demonstrated that GP Ⅱb/Ⅲa mAb significantly decreased atherosclerotic lesions and HMGB-1, TLR4 and NF-κB expression in ApoE-/- mice (P < 0.05). The present study has suggested a possibility that GP Ⅱb/Ⅲa blockade attenuates atherosclerosis by inhibiting the HMGB-1/TLR4 pathway.]]> 2013/9/16 0:00:00 GU Hong-Feng,JIANG Jian-Hong,LIAO Duan-Fang,TONG Qiao-Zhen and YANG Yong-Zong 538true FBXO38, which was designated as gFBXO38t1. Real-time RT-PCR analysis showed that gFBXO38t1 was widely expressed in various chicken tissues, with a relatively higher expression level in the pancreas, ileum and abdominal fat tissue. In addition, gFBXO38t1 was expressed in all the chicken abdominal fat tissues used in the present study, and at 3 and 4 weeks of age, the gFBXO38t1 expression in lean males was significantly greater than that in fat males (P < 0.05) and no significant difference was observed at the other ages (P > 0.05). The gFBXO38t1 expression decreased followed by the differentiation of chicken preadipocytes induced by oleate, and gFBXO38t1 expressed more highly in chicken preadipocytes than in mature adipocytes (P < 0.05), suggesting that gFBXO38t1 might play a negative role in the chicken adipogenesis. Additionally, The luciferase reporter assay showed that gFBXO38t1 overexpression inhibited the promoter activities of chicken CCAAT/enhancer-binding protein α (C/EBPα), lipoprotein lipase (LPL), fatty acid synthase (FASN) and fatty acid-binding protein 4 (FABP4). The combined overexpression of gKLF7 and gFBXO38t1 did not lead to an enhanced ability for gFBXO38t1 to regulate the promoter activities of chicken LPL, FABP4 and FASN, indicating that gFBXO38t1 may regulate promoter activities of these genes through a KLF7-independent manner. The current study provides evidence that the gFBXO38t1 is involved in chicken adipogenesis.]]> 2013/9/16 0:00:00 ZHANG Zhi-Wei,SUN Ying-Ning,RONG En-Guang,LI Hui and WANG Ning 537true p/dt) of left ventricular pressure and left ventricular systolic pressure (P < 0.05), as well as a significantly lower rate of left ventricular diastolic pressure (P < 0.05). The deposition of collagen fibers in the myocardium of transplantation group was reduced, according to A value in transplant group is less than control group (P < 0.05). The expression of VEGF and its receptor were significantly higher than those of control group, and the upregulation of VEGF and its receptors was unsynchronized, the capillary density is higher in transplantation group than control group after four weeks. Taken together, our results suggest that BMSSCs transplantation can improve dilated cardiomyopathy's heart function of rats, we found that BMSSCs transplantation prevented the fusion of collagen fibers to some extent, decrease the deposition of collagen fiber and maintained the orderly structural arrangement of myocardial tissue. BMSSCs transplantation can also promote the angiogenesis, in our study we found that capillary density increased, BMSSCs transplantation not only upregulated the expression of VEGF, but also upregulated the expression of VEGF receptors. Unsynchronized upregulation of VEGF and its receptors was observed which may promote angiogenesis continuously and steadily.]]> 2013/9/16 0:00:00 ZHOU Ying,BAI Xiao-Juan and WANG Bo 536true 2013/9/16 0:00:00 XIONG Hui,DING Ling,WANG Jing-Jing,ZHENG Chen,SONG Jin-Zhi,PAN Bin and DENG Yan-Qiu 535true + cells from AML patients, silencing of EDAG led to decreased protein stability of NPM1 protein and increased cell sensitivity to daunorubicin. Although EDAG failed to interact with NPMc+ and regulate its protein stability in normal culture condition, with leptomycin B treatment, EDAG overexpression enhanced the protein stability of NPMc+. In AML patients with NPMc+, the CD34+ cells were more responsive to daunorubicin treatment than the cells from AML with wild type NPM1, and silencing of EDAG weakly increased the sensitivity to daunorubicin. These results suggested a potential role of EDAG in chemotherapy of AML and the "escape" of NPMc+ protein from EDAG stabilization might contribute to the favorable prognosis of AML with NPMc+.]]> 2013/9/16 0:00:00 ZHENG Wei-Wei,YANG Yang,ZHANG Mei-Jiang,DONG Xiao-Ming,TANG Liu-Jun,WANG Xiao-Hui,ZHAN Yi-Qun,YU Miao,GE Chang-Hui,NING Hong-Mei,LI Chang-Yan and YANG Xiao-Ming 534true 2 respectively. Several electron microscopic techniques, including metal shadowing, negative staining and Cryo-EM, have been used to investigate the morphology of the reconstituted 30 nm chromatin fibers. Our results suggest that both histone H1 and divalent Mg2+ can help the formation of 30 nm chromatin fibers, but the resulted chromatin fibers display different topologically architectures. To investigate how the length of linker histone may affect the architecture of chromatin，we measured the diameters of the reconstituted 30 nm chromatin fibers with different nucleosome repeat lengths (NRLs) of 177 and 200 bp and found that these two classes of chromatin fibers present different diameters (P < 0.05).]]> 2013/8/21 0:00:00 SUN Da-Peng,SONG Feng,HUANG Li,ZHANG Kuo,JI Gang,CHEN Ping and ZHU Ping 533true 2013/8/21 0:00:00 LIU Jing-Tao,YUAN Xia,XU Bo,RAN Fu-Xiang,CHU Ming-Ming,JIA Xuan,CHEN Yi-Xin,WANG Zhe,LI Run-Tao and CUI Jing-Rong 532true 5 EPCs. At 3, 7 and 14 days post EPC transplantation，the adhesion, migration abilities and capillary density in calf muscles were assessed. Results indicate that ApoA significantly reduced the adhesion and migration abilities of EPCs. Furthermore, the tubule-like formation of EPCs on Matrigel gels was damaged. In vivo experiments showed the number of EPCs home to ischemic peripheral vascular, and the number of capillary vessels decreased significantly in ApoA transgenic mice. This study demonstrated that ApoA could attenuate the adhesion, migration, and homing abilities of EPCs and could impair the vasculogenesis ability of EPCs.]]> 2013/8/21 0:00:00 WANG Ren,ZHANG Kai,LI Shuang,TONG Zhong-Yi,LI Guo-Hua,ZHAO Zhan-Zhi,LIN Xiao-Long,ZHAO Yue,LIU Feng-Tao,WANG Zuo and JIANG Zhi-Sheng 531true 3H]-ryanodine binding experiments were used to detect the change of free thiols' population, the diameter and distribution of protein complex and the gating state of RyR1. Furthermore, we used photon bleaching assay to determine the electron transfer property of the modulators. The results show that the channel activators and thiol oxidants appear to be electron acceptors that affect RyR1 in the same manner, which is to decrease the free thiols on RyR1 and to increase the size of the protein crosslinking clusters. In contrary, channel inhibitors and thiol reductants appear to be electron donors and their effects on RyR1 are opposite to activators and oxidants. Therefore, we propose that the similar electron transfer properties of channel modulators results in the similar effects on RyR1 by changing the local redox environment around the active thiols, inducing changes in thiol/disulfide balance and RyR1 gating.]]> 2013/8/21 0:00:00 SONG Wei-Tao,ZHU Qian-Rong,YE Yan-Ping and XIA Ruo-Hong 530true 2013/7/25 0:00:00 WANG Wei,LI Xiao-Ling,LI Wen-Juan,YI Mei,TANG Ke,ZHENG Pan,XIANG Bo and LI Gui-Yuan 529true 2013/7/24 0:00:00 LIU Bing and WANG Yi 528true i.e., δ, κ, and μ, which all have their respective agonists. Among them, δ-OPR (DOR) agonists are especially promising pharmaceutical chemicals for their additional anti-depressant and anti-anxiety as well as organ-protective abilities. To investigate the mechanism of δ-OPRs agonists and the receptor, in the present work 102 derivatives of N-substituted spiropiperdines were studied through three-dimensional quantitative structure-activity relationships (3D-QSAR) analysis. In conclusion, PLS analysis (Q2=0.501, R2ncv=0.787, R2pre=0.780) of the optimal CoMSIA model (yielded by hydrophobic and hydrogen-bond donor fields) manifesting good inter predictive capacity, excellent inter-consistency, and outstanding predictive ability, implies the rationality of the model. At the same time, the 3D-QSAR contour map analysis results indicate that the hydrophobic group substitution of R1 is beneficial to the activity of delta opioid agonists, and the hydrophilic or hydrogen-donor substitution of R2 is also favorable. These conclusions are helpful for understanding the mechanism of DOR agonists, as well as the guiding of design and improvement of new δ-OPR agonists in the future.]]> 2013/7/24 0:00:00 GAO Wei-Min,LI Yan,ZHANG Shu-Wei and YANG Ling 527true in situ (CIS) and invasive LSCC. As a result, 1036 proteins and 102 differentially expressed proteins were identified. Among these differentially expressed proteins, some proteins are progressively upregulation, some proteins are progressively downregulation in human bronchial epithelial carcinogenic process, and the other proteins are upregulation or downregulation in a certain stage of the process of carcinogenesis. Functional analysis and subcellular localization were performed on 102 differentially expressed proteins. Functional analysis showed that these differentially expressed proteins were associated with metabolic process, apoptosis, cell proliferation, cell differentiation, signal transduction, transcription, translation, cell adhesion, immune response, and development. Western blotting and immunohistochemistry were performed to detect the expression levels of the two proteins (S100A9 and CKB) in NBE, SM, AH, CIS and invasive LSCC, which also support the findings in MS analysis. The results suggest that these differentially expressed proteins associated with bronchial epithelial carcinogenesis, and they may be early diagnostic marker of lung squamous cell carcinoma. We further study the biological function of the differentially expressed proteins, will help to elucidate the bronchial epithelial carcinogenesis mechanism, and provide new ideas for early diagnosis and pathogenesis research of LSCC.]]> 2013/6/20 0:00:00 ZENG Gu-Qing,CHENG Ai-Lan,YI Hong,LI Mao-Yu,LI Guo-Qing,LI Jian-Ling and LIAO Li 526true 2013/6/20 0:00:00 LIU Miao,WANG Wen-Fei,LIU Ming-Yao,ZHAO Jing-Zhuang,BAI Yin,GUO Mo,REN Gui-Ping and LI De-Shan 525true +/H+ exchanger 1 (NHE1) blocker, on neointimal proliferation induced by AGEs in a carotid artery balloon injury model, the rats carotid artery was balloon injured. The exemplars were collected and stained by HE. The morphology changes were observed. The intima area, media area and the ratio of area between intima and media were calculated using image analysis system. In order to explore the precise mechanism, the experiments were done on the isolated rat vascular smooth muscle cells(VSMC). Cell proliferation was assessed by [3H] thymidine incorporation. RT-PCR and Real-time RT-PCR were used to assay the cyclooxygenase-2 (COX-2) matrix metalloproteinases-2 (MMP-2) and matrix metalloproteinases-9 (MMP-9) expression; Western blot was used to assay NF-κB protein and the degradation of the inhibitor I-κBα. The in vivo results shows that neointima hyperplasia is significantly suppressed treated with cariporide in balloon-injured rats compared with AGEs treatment lonely. The in vitro study shows that cariporide dose-dependently inhibited AGEs-induced upregulation of COX-2, MMP-2 and MMP-9 expression. We also found that cariporide blocked AGEs-induced activation of nuclear factor-κB(NF-κB)and pointed out that inhibition of NF-κB was achieved by inhibiting the degradation of the inhibitor I-κBα. The results identified that NHE1 inhibitor cariporide inhibited AGEs-induced neointimal hyperplasia in rats balloon-injured model by suppressing the proliferation of VSMC and the upregulation of COX-2, MMP-2 and MMP-9 via inhibiting NF-κB activation in VSMC. These results indicated that NHE1 might be a considerable ingredient of the signal pathway in which AGEs played a key role in the processes of vascular damage.]]> 2013/6/20 0:00:00 WU Shu-Jin,YANG Qing-Shan,SONG Tao,ZHOU Shou-Hong,LIU Yu-Hui and LIU Li-Ying 524true ex vivo, there are still some shortages in these cell sources. Depending on the multipotency and infinitive reproductive capability, human embryonic stem cells (hESCs) become the new sources. However, the efficiency for deriving endothelial progenitor/ endothelial cells (EPCs/ECs) from hESCs is low. To increase endothelial induction efficiency, we used a two-stage induction protocol. By the treatment of different cytokine cocktails at different stages and the using of Matrigel matrix, we obtained about 16% CD31+KDR+ cells from hESCs. The following endothelial induction stage with EGM2 medium showed approximate 32% subgroup of EPCs/ECs. The derived cells had the capability for tube-like structure formation on Matrigel and the ability to cohere with fluorescein griffonia (bandeiraea) simplicifolia lectinⅡ. Q-PCR indicated that the induced cells express endothelial specific genes. Immunofluorescence detection indicated the expression of endothelial specific marker-CD31. Taken together, we have successfully derived EPCs/ECs from human embryonic stem cells with a modified method.]]> 2013/6/20 0:00:00 FANG Fang,ZHANG Bo-Wen,HE Li-Juan,ZHOU Jun-Nian,WANG Jing-Xue,YUE Wen,XIE Xiao-Yan and PEI Xue-Tao 523true 2013/6/20 0:00:00 YANG Chao,DAI Wei-Min,ZHANG Xiao-Mei,CHEN Hai-Xu,YANG Bo,GONG Yuan,WANG Chang-Zheng,CHE Yu-Fang and WU Ben-Yan 522true in vitro, AS is able to catalyze the synthesis of an amylose-like polysaccharide composed of only α-1,4-linkages using sucrose as the only energy source. Unlike AS, other enzymes responsible for the synthesis of such amylose-like polymers require the addition of expensive nucleotide-activated sugars. These properties make AS an interesting enzyme for industrial applications. In spite of the great application potential, industrial applications of AS were largely hampered by its low stability. In this work, the structure of Arthrobacter chlorophenolicus amylosucrase (AcAS) was modeled according to crystal structures of Neisseria polysaccharea amylosucrase (NpAS) and Deinococcus geothermalis amylosucrase (DgAS). The structural difference of the three AS were investigated for the sake of exploring the working mechanism of AcAS. The dynamics and functional motions of AcAS were also investigated by the Gaussian network model (GNM). Based on GNM, it was found that AcAS can be divided into two regions with different moving directions. Finally, the unfolding property of AcAS was studied by the iterative GNM, and several kinetically weak regions were identified on the basis of the predicted unfolding pathway. These discoveries may be helpful to the industrial development of AS.]]> 2013/6/20 0:00:00 LIU Ming,LI Ping,LIU BIN,SU Ji-Guo and WANG Cun-Xin 521true 2+ voltage-activated K+ (BK) channels in VSMCs using patch clamp technique. The results showed that the open probability of channels increased significantly during the second and third week of exercise training, then became stable in the fourth week. After the exercise training, the sensitivity of channels to voltage and intracellular Ca2+ was enhanced, and its mean opening time got longer while the mean closed time got shorter. Our results also showed that the exercise training had no effect on channel conductance. These changes in biophysical properties of BK channels after exercise training could be considered to prevent membrane depolarization and vasoconstriction.]]> 2013/6/20 0:00:00 XU Jian-Wen,XIE Jing,WANG Fa and ZHAO Hu-Cheng 520true via self-cleaving 2A sequences. The iPSCs were generated at 3～4 weeks after ASCs have been transiently transfected with the polycistronic plasmid. Then, the iPSCs were identified subsequently via the morphological observation, immunofluorescence with specific antigen, embryoid body formation in vitro and teratoma formation in vivo. The results demonstrated that the characteristics of these generated iPSCs resembled embryonic stem cells (ESCs) in terms of the expression of pluripotent markers, and the ability to differentiate into the three embryonic germ layers in vitro by embryoid body generation together with in vivo by teratoma formation after injection into immunodeficient mice. Remarkably, Southern bolt revealed that the human iPSCs were not integrated with plasmid DNA sequence. Therefore, hASCs derived iPSCs has the pluripotency using polycistronic plasmid method, which provides a useful platform for the further study of hereditary or degenerative disease therapies with the potential to bypass both the insertional mutagenesis and immune rejection barriers.]]> 2013/5/22 0:00:00 QU Xin-Jian,LIU Tian-Qing,SONG Ke-Dong,LI Xiang-Qin,GE Dan and GUAN Shi 519true 1/2 and increasing the expression of caveolin-1. In the present study, we investigated the role of caveolae and caveolin-1 in proliferation of VSMCs and whether there are interaction between the caveolin-1 and ERK1/2 in the inhibitory effect of CGRP signal pathway. VSMCs were prepared from thoracic aorta of male Sprague-Dawley rat by the classic explants method, the passage 3 ~ 10 VSMCs were used for the present study. 10% fetal bovine serum (FBS) was employed as a stimulus for the proliferation of VSMCs. β-Cyclodextrin or filipin was used to deplete cholesterol in the caveolae. Proliferation of VSMCs was estimated by methylthiazoletrazolium (MTT) assay and Flow Cytometry. Western blotting and co-immunoprecipitation were used to determine interaction of p-ERK1/2 or caveolin-1. Results showed that CGRP significantly inhibited VSMC proliferation and down-regulated phosphorylation of ERK1/2. Incubation of VSMCs with β-cyclodextrin or filipin promoted cells proliferation, up-regulated phosphorylation of ERK1/2, attenuated the inhibitory action of CGRP on VSMC proliferation and decreased caveolin-1 expression. Pretreatment with CGRP increased the direct binding of cavolin-1 with phosphorylated(p-) ERK1/2 but not non-phosphorylated ERK1/2 in the presence of 10?S. Our results revealed that caveolae and caveolin-1 may contribute to the inhibitory effect of CGRP on the VSMC proliferation, and the mechanism may be related to the deceased nuclei translocation of p-ERK1/2 because of the increased binding of caveolin-1 with p-ERK1/2.]]> 2013/5/22 0:00:00 CHEN Yun,DAI Zhong,LIU Yan-Mei,TIAN Hai-Hong,DENG Shui-Xiu,CHEN Lin-Xi,Wang H Donna and QIN Xu-Ping 518true 2013/5/22 0:00:00 ZHENG Quan-Hui,ZHANG Ai-Hong,ZHENG Ai-Hua,YAO Wen-Bo,GAO Jin-Ming,ZHANG Qing-Bo and LI Juan 517true in vivo. A possible mechanism of inflammation inhibition involving chemokine mRNA by the peptide was determined by analyzing mRNA expression levels of chemokines and tristetraprolin (TTP) by SqRT-PCR.Nineteen phage clones were selected after four rounds of biopanning with a cut-off of 3-fold higher binding to LPS-activated PBMCs than to normal PBMCs. Nine of the phage clones inhibited IL-8, MCP-1, and MIP-1β production in vitro. Five clones displayed the same peptide(CI-S5)most robustly inhibited the chemotactic activity in vitro and reduced paw oedema and arthritis in Wistar rats in vivo. SqRT-PCR results indicated that mRNA expression of IL-8, MCP-1, and MIP-1β were reduced and TTP mRNA expression was increased in the CI-S5 treatment group. Our data demonstrate that CI-S5 is a broad-spectrum antagonist of pro-inflammatory chemokines as it enhances the expression of TTP to reduce chemokine mRNA expression. This study provides a basis for the development of new peptide-based therapies for the treatment of inflammatory diseases.]]> 2013/5/22 0:00:00 SU Yan,SUN Han-Xiao,LI Xiu-Ying,MO Xue-Mei and ZHANG Guang 516true 2013/5/22 0:00:00 ZUO Zhen-Tao,LUO Huan and ZHOU Ke 515true sll1466, from Synechocystis PCC 6803. Thin-section electron micrographs showed that the mutant Synechocystis contained more carboxysomes when grown in the absence of CO32- and while more glycogen granules when grown in the presence of 0.5 mol/L NaCl than the wild type Synechocystis. Absorption spectra of the mutant grown under different light intensities differed significantly from wild type Synechocystis. At the molecular level, the mutant differed from the wild type in 3 aspects: (1) glycosylation of two carbohydrate-selective OprB porins in the thylakoid membrane; (2) phosphorylation of the rod-core linker, CpcG1 (Slr2051), in the supernatant of the thylakoid membrane; and (3) transcriptional variations of genes related to the above changes. These results suggested a profound role for Sll1466 in regulating physiology, metabolism, and energy transfer.]]> 2013/4/23 0:00:00 WANG Xing,ZHENG Bing-Hui,LIU Lu-San,DONG Liang-Liang,ZHAO Kai-Hong and ZHOU Ming 514true 2013/4/23 0:00:00 WANG Yang,JIA Lei-Min,DONG Ping and LIANG Xing-Guo 513true Xa21-mediated Xanthomonas oryzae pv. oryzae (Xoo) resistance responses by Western blotting. The results indicated that OsWRKY13, OsWRKY23 and OsWRKY71 were expressed increasingly with leaf growth till the flowering stage and their expression were declined slightly at filling stage. However, the expression of OsWRKY45 and OsWRKY53 were not detected. In the Xa21-mediated resistance response, OsWRKY45, OsWRKY53 and OsWRKY71 proteins were induced, while the expression of OsWRKY13 and OsWRKY23 proteins were stable. Comparison among the resistance reaction, susceptible reaction and mock treatment revealed that the expression of OsWRKY45, OsWRKY53 and OsWRKY71 proteins had a similar pattern between resistance and susceptible reactions. This result suggested that OsWRKY13 and OsWRKY23 may play roles in rice leaf growth, OsWRKY45 and OsWRKY53 may play roles in rice-Xoo interactions and OsWRKY71 is functioning in both situations.]]> 2013/4/23 0:00:00 XU Wen-Jing,MIAO Liu-Yang,LI Li-Yun,LIU Zhao,LIU Yu-Meng,JIANG Guang-Huai,YANG Feng-Huan,HE Chen-Yang and LIU Guo-Zhen 512true in vivo extracellular recording, the present study investigated the intensity-tuning properties in AI neurons of mouse (Mus musculus, Km). We also examined the effect of cortical application of the GABAa receptor antagonist bicuculline on AI intensity tuning in order to indentify the possible source of inhibition. The intensity-tuning curves were recorded in 72 AI neurons among which 28 showed monotonic responses and 44 showed non-monotonic responses. In non-monotonic neurons, there was no change in the first spike latency but a decrease in the firing duration (P < 0.01) when the sound intensity increased from best intensity to highest intensity. After AI application of bicuculline by electrophoresis (n = 28), the non-monotonic responses did not change in 39.3% neurons, was weakened in 42.9% neurons and was enhanced in 17.9% neurons. It suggested that GABAergic inhibition must be involved in AI intensity tuning but was not the only source. Non-monotonic excitatory input and non-GABAergic inhibition should also be considered. A postulated model was presented to explain the mechanism of intensity-tuning in AI neurons.]]> 2013/4/23 0:00:00 QI Qiao-Zhen,SI Wen-Juan,LUO Feng and WANG Xin 511true in vivo, and decreasing immunogenicity. The effects of pH, reaction time, protein concentrations and mass ratio between the reactants on the PEGylation of mFGF-21 were analyzed. PEG-mFGF-21 was isolated by Capto Q anion exchange chromatography or Superdex 75 gel filtration chromatography. As a result, the optimal reaction method for mFGF-21 PEGylation and purification processes of PEG-mFGF-21 were established. Then we study the physical and chemical properties, immunogenicity, in vivo half-life, in vitro biological activity and in vivo hypoglycemic effects of PEG-mFGF-21. We found that the temperature stability and anti-protease ability of mFGF-21 were significantly improved after PEGylation. The indirect ELISA results for detection of serum antibody levels against mFGF-21 and the target protein concentration showed that PEGlyation of mFGF-21 significantly reduced its immunogenicity and increased its half-life in vivo. The glucose uptake assay results in HepG2 cells demonstrated that the in vitro activity of PEG-mFGF-21 did not decline. However, with the increment of stimulating time, the glucose uptake in cells treated with PEG-mFGF-21 significantly increased than mFGF-21. The experimental results of short-term blood glucose regulation in type 2 diabetic animals showed that the hypoglycemic speed of mFGF-21 was faster than PEG-mFGF-21, but it lasted shorter. The long-term blood glucose regulation experimental results showed that PEG-mFGF-21 had better hypoglycemic effect than mFGF-21. Surprisingly, blood glucose of PEG-mFGF-21-treated mice remained at low level for several days after the drug was withdrawn. In conclusion, under the premise of remaining its biological activity in vitro, PEGylation of mFGF-21 can improve its physical stability and anti-protease ability, decrease its immunogenicity and increase its in vivo half-life. It can also prolong the hypoglycemic effect of mFGF-21 in diabetic animals. This study provides an important technology platform for drug development of FGF-21.]]> 2013/4/23 0:00:00 YE Xian-Long,ZHAO Jing-Zhuang,REN Gui-Ping,YU Dan,LIU Ming-Yao,YU Yin-Hang and LI De-Shan 510true 2013/3/26 0:00:00 LIN Yun,LI Zhi-Hui,WANG Ran,LI Wen-Juan,WANG Jing-Jing,JI Chao-Neng,XUE Jing-Lun and CHEN Jin-Zhong 509true 60Co γ-rays at the rate of 100 cGy/min. (B) i.v. transplanted with physiological saline, (C) i.v. transplanted with MSCs. Mice were weighed daily and observed for 15 days after irradiation. Intestines of mice were obtained in order to assay histological changes by hematoxylin and eosin staining. The immunofluorescence staining showed that the transplanted cells could differentiate into vimentin+/α-SMA+ fibroblastic-like cells in vivo. These data indicated that MSCs could engraft at the injured small intestine, and contribute to the repairment of irradiated intestine. This could provide a valuable strategy for radiation-induced enteritis.]]> 2013/3/22 0:00:00 YANG Chao,ZHANG Xiao-Mei,GONG Yuan,WANG Chang-Zheng,HAN Ying and WU Ben-Yan 508true 642～1290) with double-stranded DNA (dsDNA) by fluorescence polarization technology, and analyzed the related characteristic parameters to understand the DNA-binding and unwinding properties of BLM642～1290 helicase. The results indicated that the binding and unwinding of the helicase and dsDNA was related to the length of 3'-tailed single-stranded DNA (ssDNA) in dsDNA substrates; the helicase preferred to bind the ssDNA tails of dsDNA substrates, and every helicase molecule may bind 9.6 nt ssDNA; the unwinding efficiency was the highest level when the 3'-tail ssDNA length of dsDNA substrates was 10nt, no longer increasing with the length. In addition, the blunt dsDNA was also bound and unwound by BLM642～1290 helicase, but the binding affinity and the unwinding efficiency were smaller than 3'-tailed dsDNA. It may be concluded that BLM642～1290 helicase was monomer in binding and unwinding to dsDNA substrates, and may unwind dsDNA in an inchworm manner. These results will provide the theoretical foundation for further studying the unwinding mechanism of BLM helicase.]]> 2013/3/22 0:00:00 LUO Heng,XU Hou-Qiang,CHEN Xiang,LIU Chao-Qian,XU Qing-He and LI Kun 507true 2013/2/26 2:47:13 ZHAO Jing,BI Hong-Yan and QIAN Yi 506true in vitro and in vivo. Moreover, bioinformatics analysis, flow-cytometric analysis and PCR array analysis were performed to elucidate the mechanism under the role of ZNF403 in cell growth. Knockdown of ZNF403 significantly decreased the rate of DNA synthesis and mitosis. Additionally, a number of key cell-cycle regulating components such as MCM2, p21, ATM and MRE11A were identified to be mediated by ZNF403. Altogether, our findings suggest that ZNF403 is a novel cell cycle regulator, which may play an essential role in tumorigenesis.]]> 2013/2/26 2:55:46 GUAN Rui,HOU De-Fu,RAO Xiang,GUAN Yong-Jun,OUYANG Yong-Mei,YU Yan-Hui,Jim HU and CHEN Zhu-Chu 505true in vitro was detected on breast cancer cell line MDA-MB-231 cells transducted with miR-132 (miR-132 group) and or mock miRNA (control group) by transwell assay. The results demonstrated significantly lower migration ratio in miR-132 case compared to that in control case, indicating inhibition effects on cancer migration of miR-132. To clarify the inhibition mechanism by which miR-132 inhibits cancer metastasis, target genes of miR-132 were screened and identified. They are CHIP (STUB1), G3BP1 and G3BP2. The expression levels of these 3 genes in MCF7 cells (metastasis cell line) and MDA-MB-231 cells with or without transduction of miR-132 or mock miRNA were detected by PCR and Real-time PCR. Two key genes, G3BP1/G3BP2, were founded to be involved in the regulation of miR-132 to tumor metastasis, demonstrating that miR-132 could silence G3BP1/G3BP2, which resulted in the suppression of tumor metastasis. Our research suggests that miR-132 may be an important potential target used for inhibition of cancer metastasis and clinical therapy of cancer, and shed light on the suppression mechanisms of miR-132.]]> 2013/2/26 3:06:13 GUAN Di,LIU Chun-Ying,CHEN Chen and YIN Qin-Wei 504true 2013/2/26 0:00:00 LIU Shuai,NING Xiao-Min,LI Mei-Hang,QIU Yang,LI Yan-Jie,DONG Pei-Yue and YANG Gong-She 503true 2013/2/27 2:37:08 LI Wen,LI Wen-Ting,CAO Shu-Juan,YU Wen-Xiang,LI Wen-Jie,DAI Jun-Biao and SUN Zhi-Rong 502true 2013/2/27 0:00:00 WANG Jing-Jing,WANG Ran,LI Wen-Juan,XUE Jing-Lun and CHEN Jin-Zhong 501true 2013/2/27 0:00:00 ZHAO Xiao-Meng,WANG Cheng,LI Xiao-Feng,ZHANG Xiao-Ting,LIU Xi-Zhi and ZHOU Chang 500true via the Wnt signal pathway, and the Wnt signal pathway might be used as a new target for protecting pancreatic β cells against cytokine-induced cell dysfunction and death.]]> 2013/2/27 0:00:00 DING Ye,DAI Xiao-Qian,ZHANG Zhao-Feng and LI Yong 499true in vitro and three germ layers in vivo. After the cells were used as nuclear donors, the cleavage rate of the cloned embryos was 53.33%, the morula rate 9.03%, the blastocyst rate 2.07%, and the total cell number per hatched blastocyst was 26.5. Compared with embryos cloned from non- lentivirus PFBs, the morula rate and blastocyst rate were lower and significantly different (P < 0.05). Lentivirus can result in the generation of porcine iPS cells from PFBs, so it can be used as ideal material and tool for research such as epigenetic modification and cell reprogramming.]]> 2013/2/27 0:00:00 CAO Hong-Guo,CHEN Tao,YIN Hui-Qun,SUN Xue-Ping,PU Yong,YANG Pan,ZHANG Yun-Hai,LIU Ya,LI Yun-Sheng,TAO Yong and ZHANG Xiao-Rong 498true ZNF185 was cloned and transfected into K562 cells, MTT assay showed that over expression of ZNF185 in K562 cells significantly inhibited its proliferation. To explore the molecular mechanism that regulates ZNF185 expression in myeloid leukemia cells, methylation specific PCR (MSP) was performed to determine the methylation status of ZNF185 promoter in the normal PBL neutrophils, HL-60 and K562 cells. Higher methylation of ZNF185 promoter was detected in HL-60 and K562 cells than that in normal PBL neutrophils. Furthermore, treatment of K562 cells with 5-aza-CdR, a DNA demethylation agent, resulted in demethylation of ZNF185 promoter, increased ZNF185 expression and inhibited K562 cell proliferation. Our results indicate that ZNF185 promoter methylation which reduces ZNF185 expression in K562 cells and its ability to inhibit cell proliferation may be one of the molecular mechanisms that are related to the occurrence or development of chromic myeloid leukemia.]]> 2013/2/28 10:12:14 ZHENG Quan-Hui,ZHANG Ai-Hong,ZHENG Ai-Hua,YAO Wen-Bo,ZHANG Qing-Bo,LU Bin,HU Xue-Lian,WANG Na and ZHAO Yong 497true Escherichia coli (E. coli) is easily cultured and genetically manipulated, thus is potentially an excellent display host, the display technology based on E. coli has not been widely used. One of the problems is lack of efficient display of antibodies on the surface of E. coli. Many proteins have been tested as display carriers in outer membrane display in E. coli. Display systems based on autotransporter protein (AT) and ice nucleation protein (INP) is the most extensively studied. Another problem is unstable survival rates of E. coli when antibody is displayed. In this study, we systematically examined display level, antigen-binding affinity of displayed antibody and survival rate of E. coli using Ag43β(β domain of Antigen43, an AT protein) and INPNC (fragment of N-terminal and C-terminal of INP) as carrier proteins and T7, lac, araBAD as promoters for antibody expression. We found that the antigen-binding ability of the Ag43β based display was superior to that of the INPNC based system. As expected, T7, lac and araBAD promoters drove high, medium and low expressions of antibody. The host survival rate using T7 promoter was extremely low (INPNC: 0.0033%, Ag43β: 0.02%, the host bearing araBAD promoter had the highest survival rate (INPNC: 37.80%, Ag43β: 90.23%), and the lac based system had a survival rate of 2.04% (INPNC) and 13.27% (Ag43β). Balancing the antigen-binding abilities, antibody expression levels and survival rates, a system using Ag43β as carrier protein and lac as promoter is the best choice for antibody display on E. coli.]]> 2013/12/20 0:00:00 YANG Xiao,SUN Shuang,WANG Hai-Feng and HANG Hai-Ying 496true in vitro and in vivo assays were conducted to detect whether BMP9 can induce osteogenic differentiation of MSCs through JNKs kinase pathway. The results showed that BMP9 can activate JNKs kinase through increase the phosphorylated form of JNKs kinase. JNKs kinase inhibitor SP600125 can inhibit ALP activity, OPN and OCN expression, as well as calcium deposition induced by BMP9 in MSCs. Furthermore, SP600125 also led to a decrease in BMP9-induced Runx2 activity and canonical Smad signaling. Moreover, when JNKs kinase was silenced by RNA interference in MSCs, BMP9-induced osteogenic differentiation in vitro and ectopic bone formation in vivo were accordingly inhibited along with knockdown of JNKs. Taken together, those results intensively suggested that BMP9 can induce and regulate osteogenic differentiation of MSCs through activating JNKs kinase pathway.]]> 2013/12/20 0:00:00 XU Jing,ZHAO Dan,WANG Jian,WANG Wen-Juan and LUO Jin-Yong 495true via ubiquitin Lys-48. The HeLa cells were co-transfected with the same quantity of the CD9 plasmids and the gradient increase quantity of the RNF149 plasmids. We found that the exogenous quantity of CD9 was decreasing with the increased expression of the exogenous RNF149. In HEK293T cells, the knocking down RNF149 by shRNA led to the increase of the endogenous CD9. All these evidence suggested that CD9 maybe regulated by RNF149. In addition, the knocking down RNF149 by shRNA led to the inhibition of the cell proliferation in HEK293T cells. This phenomenon suggested that the RNF149 possibly could be considered as the regulatory factor of the cell proliferation.]]> 2013/12/20 0:00:00 LI Yan,RUAN Lin-Hao,SUO Ta-Lin,WANG Peng and TANG Jie 494true etc, CGTase has become the focus of scientific research nowadays. Calcium binding sites widely exit in α-amylase family. Previous studies indicated that these sites had very important roles for α-amylase. It was known that CGTases also possess two or three calcium binding sites. However, their structure and function are not very clear. In the present study, structure and function of calcium binding sites in CGTases were analyzed. Sequence comparisons were performed using the ClustalX 1.8 sequence alignment program. Based on the results and crystal structure analysis, it was found that calcium binding sites CaⅠ and CaⅡ exist commonly in CGTase. Most amino acids at these two calcium binding sites are highly conserved, but the residue 29 at CaⅠ and residue 199 at CaⅡ have significant differences between different types of CGTases. The residue 29 in α-CGTase primarily producing α-cyclodextrin or γ-CGTase primarily producing γ-cyclodextrin is Asp, while others are Asn. The residue 199 in γ-CGTase is Ser, while others are Asp. Calcium binding site CaⅢ only exists in few CGTases. The site consists of residues 315 and 577. In addition, site-directed mutagenesis was used to investigate the functions of calcium binding sites in CGTases. The replacement of Asp29 by Asn and Arg resulted in 23% and 35% increase in β-cyclization activity, respectively. Mutant D29R and D315A showed higher stability than wild-type CGTase at 60℃. Moreover, the mutant D315A had higher β- and γ-cyclodextrin specificity. These results suggested that calcium binding sites might be related to cycling activity, thermal stability, and product specificity of CGTases, which provided the directions for further revealing biological functions of calcium binding sites of CGTases.]]> 2013/12/20 0:00:00 BAN Xiao-Feng,LI Cai-Ming,Bao Chun-Hui,Gu Zheng-Biao and Li Zhao-Feng 493true 2013/12/20 0:00:00 WANG Peng-Yun,LI Juan,LI Hui-Jie,LI Bing,LI Xin and ZHOU Ting 492true Pseudomonas aeruginosa. Las and Rhl are two major quorum-sensing components, and they are regulated by their corresponding autoinducers, N-3-oxododecanoyl homoserine lactone (3-oxo-C12-HSL) and N-butyryl-L-homoserine lactone (C4-HSL). Recent progress has demonstrated the potential of quorum-sensing molecules, especially 3-oxo-C12-HSL, for modulation of the host immune system. Here we show the specific ability of 3-oxo-C12-HSL to induce apoptosis in mice-derived macrophages (RAW264.7) and reduce phagocytosis of the cell. When RAW264.7 cells were incubated with synthetic 3-oxo-C12-HSL, the significant loss of viability was observed in a concentration (6.25 to 100 μmol/L) and incubation time (2 to 24 h) dependent manner. The cytotoxic activity of 3-oxo-C12-HSL was also observed in RAW264.7 cells. The cells treated with 3-oxo-C12-HSL revealed morphological alterations indicative of apoptosis. Acceleration of apoptosis in 3-oxo-C12-HSL-treated cells was confirmed by multiple criteria (caspases 3, 8 and 9, mitochondrial depolarization, phosphatidylserine expression). Phagocytosis of neutral red assay demonstrated that higher concentration of 3-oxo-C12-HSL significantly reduced phagocytosis of RAW264.7 cells (P < 0.05). High concentration of 3-oxo-C12-HSL also obviously lowered RAW264.7 cells for gobbling up of P. aeruginosa capability (P < 0.001). These data suggest that 3-oxo-C12-HSL specifically promotes induction of apoptosis and reduces the phagocytosis of RAW264.7 cells, which may be associated with 3-oxo-C12-HSL-induced cytotoxicity in RAW264.7 cells. Our data suggest that the quorum-sensing signal molecule 3-oxo-C12-HSL has critical roles in the pathogenesis of P. aeruginosa infection, not only in the induction of bacterial virulence factors, but also in the modulation of host responses.]]> 2013/11/22 0:00:00 YANG Wang,CHEN Song,YANG Zhen-De,XIAO Xin and LIAO Fang 491true 33-35 in C57BL/6 mice was performed. From the clinical course, we found no significant difference between male and female mice in the incidence and onset time, but males had a more serious disease score than that in females. And histological examination of spinal cord was performed at day 21 post-immunization. Compared with male mice, female mice had a dramatic increase of leukocyte infiltration in the spinal cord. Luxol fast blue staining also revealed enhanced demyelination in male mice compared to female mice. The CNS leukocyte infiltration was also quantified by flow cytometry analysis at day 19 post-immunization. The results again confirmed that the CD4 T cells accumulated in the CNS of EAE mice were increased in male mice. We further quantified the number of TH-1 and TH-17 cells in the CNS infiltrates and found that the absolute number of both TH-17 and TH-1 cells were significantly increased in male mice. Taken together, these data indicate that sex difference exists in EAE induced by MOG33-35 in C57BL/6 mice, and this will provided useful indications for further studies on the selection of animals in the EAE model.]]> 2013/11/22 0:00:00 WANG Ting-Ting and DU Chang-Sheng 490true FMR1)gene, which is widely expressed in cells of the nerve, muscle, and testes. Fragile X related protein 1 (FXR1P) is encoded by a homologous gene to FMR1——Fragile X related gene 1 (FXR1) and can interact with proteins and RNAs. Many illnesses were involved in the altered expression of FXR1. To understand the biological effect of the interaction between FXR1P and CMAS, we constructed a FXR1 overexpression vector and investigated its expression in PC12 (the rat pheochromocytoma) cells and VSMC (vascular smooth muscle cell) and the effect of the overexpression on cell morphology and several cell processes related to CMP-N-acetylneuraminic acid synthetase (CMAS) activity. We demonstrate that the overexpression of FXR1 gene can increase activity of CMAS in PC12 cells and provide a certain degree growth protection for that cells. Thus, it suggests FXR1P is a tissue-specific regulator to alter the concentration of GM1 in PC12 cells, but not in VSMC.]]> 2013/11/22 0:00:00 MA Yun,QIN Ling-Xue,DONG Xiao,LI Bin-Yuan,WANG Chang-Bo,XU Can,WANG San-Hu and HE Shu-Ya 489true P < 0.01). However, influenza virus growth were not inhibited with Geldanamycin on 36 h and 48 h(P > 0.05)after infection with influenza virus H5N1. Analysis of proinflammatory cytokines in Geldanamycin-treated A549 cells revealed significant reductions in IFN-α，TNF-α or IL-6 on 12 h and 24 h after infection with influenza virus H5N1(P < 0.05). All the results indicated that Geldanamycin have the dual effect of combine antivirus with anti-inflammation, which provided the science data for the potential use of Geldanamycin as a novel, highly effective anti-influenza virus drug.]]> 2013/11/22 0:00:00 HU Yi,LIU Peng-Peng,ZHANG Zhi-Ming,LU He-Zhen,LUO Jing,WANG Cheng-Min,LIU Jun-Xu,ZHAO Bao-Hua and HE Hong-Xuan 488true PR) genes were known to be induced by pathogen invasion. Immerging evidences revealed that the transcription of PR genes also up-regulated under abiotic stresses, however, limited data for PR proteins expression was reported. To characterize the expression of rice PR proteins under stresses, we examined the expressions of eight PR proteins under cold, hot, drought, submerge and salt stresses at seedling stage by Western blotting (WB) via antibody-based proteomics strategy. The results showed that the expressions of PR8 was up-regulated under cold stress, the expressions of PR1a, PR3, PR5 and PR16 were down-regulated under hot stress. The expressions of PR1a, PR2 and PR8 were up-regulated while PR5 and PR16 were down-regulated under drought stress, the expressions of PR1, PR2 and PR15 were up-regulated and PR1a, PR3, PR5 and PR8 were down-regulated under submerge stress. The expressions of PR2 and PR3 were up-regulated while PR1a, PR5, PR8 and PR16 were down-regulated under salt stress. In addition, a number of stress responsive cis-elements, such as the ABRE, TC-rich repeats and HSE, were identified in the promoter region of PR genes. These data supported that PR proteins play important and specific roles in the process of stress tolerance.]]> 2013/11/22 0:00:00 GAO Qing-Hua,ZENG Xiang-Ran,JIA Lin,NIU Dong-Dong,LI Xue-Jiao,GUAN Ming-Li,JIA Meng,LAN Jin-Ping,DOU Shi-Juan,LI Li-Yun,LIU Li-Juan and LIU Guo-Zhen 487true E. coli, FabA, a bifunctional enzyme, is the key enzyme of the classic anaerobic pathway of unsaturated fatty acid synthesis and introduces the cis double bond into a 10-carbon intermediate. This intermediate is then elongated by FabB (one of long chain 3-ketoacyl-ACP synthases) to form the unsaturated fatty acids found in the membrane phospholipids. Sequence alignments indicated that Sinorhizobium meliloti SmFabA and SmFabB are 60.6% and 61.1% identical to E. coli FabA and FabB, respectively. Further analysis showed that the conservative active-site histidine residue in EcFabA and the Cys-His-His catalytic triads in EcFabB, are also found in SmFabA and SmFabB. The genetic complementary revealed that SmfabA is able to restore the growth and the fatty acid synthesis of the E. coli temperature sensitive mutant CY57 at nonpermissive temperature under addition low concentration triclosan to inhibit enoyl-acyl carrier protein reductases. Moreover, SmfabB is able to complement temperature sensitive mutant CY242. In vitro assay identifies that SmfabA, like E. coli FabA, is able to introduce the cis double bond into a 10-carbon intermediate, and SmfabB, like FabB, is able to condense the acyl-ACPs with malonyl-ACP to long-chain acyl-ACPs. However, we also attempted to inactivate the fabA and fabB genes by allelic replacement but none of the fabA and fabB deletion mutant was obtained, and it seemed likely that fabA and fabB are essential genes in S. meliloti. These results demonstrated that SmFabA and SmFabB are key enzymes in unsaturated fatty acid synthesis in S. meliloti.]]> 2013/11/22 0:00:00 HU Zhe,MA Jin-Cheng,JIANG Jing-Jing and WANG Hai-Hong 486true 2013/11/22 0:00:00 XU Bing,HAN Jin-Duo,WANG Ao-Jin and HU Kun-Sheng 485true Populus tomentosa 4CL1 (Pt4CL1) complexed with its enzymatic intermediate 4-coumarate-AMP anhydride. The Pt4CL1 was found to adopt the conformation of the second step reaction based on comparisons with other members from the same superfamily. Structural analysis revealed that the residue His-234, which forms a hydrogen bond to the phosphate group of the 4-coumarate-AMP anhydride through its side chain imidazole group, plays essential roles in the enzymatic mechanisms of Pt4CL1, including facilitating both steps of the reactions by forming a hydrogen bond to, and reducing the negative charge of, the phosphate group; de-protonating the thiol group of CoA; and regulating the access of the active site to CoA through the conformational changes of its side chain. Enzymatic assays on Pt4CL1 and relevant mutant verified that His-234 is an essential residue of this enzyme.]]> 2013/11/22 0:00:00 REN Bai-Guang,LI De-Feng,ZHENG Cai-Xia,GAI Ying,JIANG Xiang-Ning and HU Yong-Lin 484true 487SPKK490" located at the dimer interface. We demonstrated that the phosphorylation-mimetic mutation of Ser487 leads to the dissociation of the CC1-FHA dimer. Moreover, the Ser487-mutation could regulate the motor activity of KIF1A and the KIF1A-mediated axonal transport activity of SVs in C. elegans. Thus, the highly conserved "487SPKK490" motif may be a key site in the CC1-FHA tandem for regulating CC1-FHA dimerization and the subsequent activity of KIF1A.]]> 2014/9/22 0:00:00 LIU Bei,YUE Yang,YU Yong,REN Jin-Qi,FENG Wei,HUO Lin and XU Tao 483true v3 channels are fast inactivating rate and strong voltage-dependence. We have investigated the molecular basis for determining the voltage-dependence of inactivation for Cav3.1, focusing on domain Ⅰ and Ⅱ. We made chimeras between Cav3.1 and Cav1.2. Chimeras were expressed in oocytes and currents were recorded by voltage clamp. For domain Ⅰ, replacement of S1～S4 or S5～S6 shifted the steady state inactivation curve significantly. These changes were mainly or partially caused by activation-inactivation coupling, rather than molecular modification. Replacement of domain Ⅱ shifted the inactivation curve significantly and these changes refer to molecular modification, indicating that domain Ⅱ contributed to the voltage-dependence of inactivation for Cav3.1. Furthermore, both voltage sensor region S1～S4 and pore region S5～S6 in domain Ⅱ were also involved, but Ⅰ-Ⅱ linker has no contribution. In addition, we found that the Ⅰ-Ⅱ linker and S5～S6 in domain Ⅰ contributed strongly to inactivation rate for Cav3.1, while S1～S4 in domain Ⅰ and Ⅱ was not involved. Taken collectively, our results suggest that domain Ⅱ plays a key role in determining the voltage-dependence of inactivation for Cav3.1, which was different from the molecular determinants for inactivating rate and for voltage-dependence of activation.]]> 2014/9/22 0:00:00 HE Bing-Jun,HU Fen,SHANG Xue-Liang,HAN Li-Xin,WU Guang-Yan,LI Jun-Ying and SUN Jin-Sheng 482true Escherichia coli. In addition to participating synthesis of long acyl chains, FabB is a key enzyme responsible for a condensation reaction in de novo unsaturated fatty acid synthesis and to form palmitoleoyl-ACP. While the FabF was found to be required for the elongation of cis-9-hexadecenoyl-ACP to cis-11-octadecenoyl-ACP and not involved in de novo unsaturated fatty acid synthesis. It has been reported previously that FabF homologues were found to have both KAS Ⅰ and Ⅱ activity just like FabB and FabF of E. coli in Enterococcus faecalis, Lactococcus lactis, Clostridium acetobutylicium and Ralstonia solanacearum. To test if this phenomenon is prevalent, we have carried out functional identification of five fabF homologous genes, Bacillus subtilis BsfabF, Sinorhizobium meliloti SmfabF, Vibrio cholera VcfabF, Pseudomonas aeruginosa PafabF1 and PafabF2. Our data revealed that five FabF homologues all have the long-chain 3-ketoacyl-ACP synthase activities in vitro. Analysis of phospholipid compositions show that SmfabF, VcfabF, PafabF1 and PafabF2 possessed 3-ketoacyl-ACP synthase Ⅱ (FabF) activity when complemented the E. coli fabF mutation CL28. The results of genetic complementation and thin-layer chromatographic analysis show that only PafabF2 gene could complement E. coli fabB mutation and PaFabF2 possessed partial function of 3-ketoacyl-ACP synthaseⅠ(FabB). These results demonstrated that not all FabF homologues have dual activity of KAS Ⅰ and KAS Ⅱ.]]> 2014/9/22 0:00:00 MA Jin-Cheng,DENG Li-Ting,TONG Wen-Hua,ZHU Lei and WANG Hai-Hong 481true Chaetoceros curvisetus(C. curvisetus) and Karenia mikimotoi (K. mikimotoi), were chosen to study the effects of temperature on microalga growth, nitrate reductase activity (NRA), and their relationship under laboratory set-up. At the beginning of the incubation, DIN, PO4-P, and SiO3-Si were added to 4 500 ml sea water in 5 L bottles to form a final concentration of 32 μmol/L, 1.5 μmol/L and 32 μmol/L, respectively. The bottles were then incubated under ambient sunlight(about 50 W·m-2) at different temperatures (10℃, 15℃, 20℃, 25℃, and 30℃) for 10～20 days. Triplicates were employed for all treatments. The microalgae cell numbers were counted by hemacytometer counting method. S-logistic 2 population growth model was used to simulate microalgae growth , and calculate two growth-related parameters, the maximum growth rate (μmax) and the final biomass (Bf). The activity of nitrate reductase was measured in vitro and its relations to microalgal growth were elucidated. The two microalgae were observed to adapt to different temperature ranges. C. curvisetus could grow normally in the range of 10℃～30℃, while K. mikimotoionly grew well between 15℃～25℃. Both μmax and Bf reached the peak values at 20℃ for C. curvisetus and at 25℃ for K. mikimotoi, respectively. These results indicated that compared to C. curvisetus, K. mikimotoi had a relatively narrow range of suitable temperature and a high optimum temperature (Topt). Throughout the incubation time, NRA of the two species changed similarly under different temperatures. NRA raised slowly at the first 3～5 days, and then dropped rapidly. The maximum NRA values (NRAmax) usually occurred at the early exponential growth phase, prior to appearing time of μmax and Bf. The relations of μmax, NRAmax and Bf with temperature followed the Shelford tolerance law. All above results suggested that temperature could affect the nitrate assimilation ability of microalgae, and subsequently affect the growth of algae. μmax and the NRAmax per unit volume of the C. curvisetus were higher than those of K. mikimotoi under the same environmental conditions, indicating that C. curvisetus might have a better ability of absorbing and utilizing nitrate for self-growth than K. mikimotoi.]]> 2014/9/22 0:00:00 DAI Ai-Quan,SHI Xiao-Yong,DING Yan-Yan,TANG Hong-Jie,WANG Li-Sha and WANG Xiu-Lin 480true 2014/9/22 0:00:00 NIU Ge,MA Juan,LIU Chang-Zhen,MA Pan,HAN Hua-Min,ZHAO Yun-Feng,ZHANG Ge,TAO Hua and GAO Bin 479true E. coli. Pull-down assay was carried out by using the purified SelW′, and exogenously direct interaction between SelW′ and PSAP was also verified. Finally, co-IP method was applied to successfully verify the endogenous interaction between the two proteins in the brain tissues of Kunming mice. All these results show that SelW interacts with PSAP directly both in vivo and in vitro. SelW may play a key role in the brain development and neurodegenerative disease formation by interacting with PASP.]]> 2014/8/19 0:00:00 CHEN Ping,LIU Qing,MA Xiao-Jie,WANG Shi-Jie,LIU Qiong and NI Jia-zuan 478true -/-, ABCG4-/- and ABCG1-/- ABCG4-/- double knockout mice. ABCA1 mRNA expression were detectable in multiple tissues, and ABCG1, ABCG4 were highly expressed in adult brain. ABCG1 and ABCG4 mRNA levels peaked at 42 days of age, while ABCA1 mRNA levels were near baseline. ABCG1 protein levels peaked at day 28 then decreased, while ABCG4 levels peaked at day 42. ABCA1 levels remained near baseline. Interestingly, circulating plasma and brain esterified cholesterol levels exhibited a biphasic distribution, which peaked at day 42. Loss of ABCG1 is compensated by increased ABCG4 and vice versa. Loss of both ABCG1 and ABCG4 results in altered expression of cholesterol synthesis related genes and cholesterol accumulation in the brain. The data suggest that ABCG1 and ABCG4, but not ABCA1 are important for transporter function in the developing brain. ABCG1 and ABCG4 play complementary roles in maintaining brain cholesterol homeostasis.]]> 2014/8/20 0:00:00 SUN Hai-Mei,CHEN Li and MIAO Jin-Wei 477true E. coli is well studied in fatty acid synthesisⅡ. FabA, a bifunctional enzyme, is the key enzyme of the classic anaerobic pathway of unsaturated fatty acid synthesis. Sequence alignments indicated that Lactococcus lactis lacks homologues of E. coli FabA, however it owns two homologues of E. coli FabZ, LlfabZ1 and LlfabZ2. LlFabZ1 and LlFabZ1 are 41% and 45.1% identical to E. coli FabZ, respectively. Further analysis showed that the conservative active-site in E. coli FabZ is also found in LlFabZ1 and LlFabZ1. Although the genetic complementary revealed that LlfabZ1 and LlfabZ2 were not able to restore the growth and the fatty acid synthesis of the E. coli temperature sensitive mutant CY57 at nonpermissive temperature, cell-free extract data showed LlFabZ1 was able to catalyze the isomerization of the trans-2-double bond to the cis-3 species. Moreover, LlfabZ2 was able to complement the EcfabZ knock out mutant HW7. In vitro assay identified that LlFabZ1 was able to introduce the cis double bond into a 10-carbon intermediate, and LlFabZ2 was able to dehydrated 3-hydroxyacyl-ACP to trans-2-decenoy-ACP. However, we also attempted to inactivate the fabZ1 and fabZ2 genes by allelic replacement but none of the fabZ1 and fabZ2 deletion mutant was obtained, and it seemed likely that fabZ1 and fabZ2 are essential genes in L. lactis. These results demonstrated that LlFabZ1 and LlFabZ2 are key enzymes in fatty acid synthesis in L. lactis.]]> 2014/8/19 0:00:00 MA Jin-Cheng,LUO Biao,HU Zhe and WANG Hai-Hong 476true C. elegans by observing worm defecation behavior. A series of genes that function in the intestine to regulate DCV biogenesis or exocytosis were successfully identified, including CAB-1, which was further determined to be a specific regulator for DCV exocytosis. In the intestine, cab-1 mutation causes reduced secretion of intestinal DCV cargoes. In the nervous system, the loss of CAB-1 leads to the accumulation of DCV markers in the presynaptic region, but synaptic vesicles are not affected. This work demonstrates that CAB-1 is a regulatory factor specifically involved in DCV secretion.]]> 2014/8/19 0:00:00 XIA Zhi-Ping,CHEN Yan,SHENG Yi,YI Ya-Lan,SONG E-Li and XU Tao 475true 2+ from cytoplasm into sarcoplasmic reticulum against the Ca2 concentration gradient. By this means, the Ca2 concentration in cytoplasm may decrease and contracted muscle cells relax. SERCA1a is the structurally and functionally best studied representative of the P-type ion transporting ATPase. Studies of SERCA1a may provide with enlightening information for research about other SERCA isoforms and P type ATPase. To understand the functional roles of the linker between the transmembrane helix M7 and the transmembrane helix M8 (L78), mutational studies about some L78 amino acid residues were performed to evaluate the effect of their single substitutions on the SERCA1a reaction cycle. Mutant SERCA1a cDNAs were obtained by Quick Change site directed mutagenesis. SERCA1a wild type and mutant proteins were expressed in COS-1 cells separately and extracted by microsome preparation. Single mutant SERCA1a proteins, wild type SERCA1a proteins and control microsomes were checked with radioactive [γ-32P]ATP, 45CaCl2, and 32Pi independently to determine their ATPase activities, Ca2 transport rates, amount of total EP formed from ATP, and amount of E2P formed from Pi. Results showed that all single mutants except for G864A transported Ca2 at lower rates than that of the SERCA1a wild type protein. Single mutations of G862 or P863 lead to severe decreases of the ATPase activity and the amount of EP formed from ATP or Pi. Measurements about ATP hydrolysis and EP formation of A893P gavde ata similar to those obtained from G862 P863 single mutant proteins, whereas single mutants of G864 and FMQ873-875 did not induce significant reduction of ATPase activity and EP amount. Experimental results above indicated that L78 is involved in the Ca2 transport across the endoplasmic reticulum membrane, and moreover, appropriate turns at GPG862-864 and near A893 are essential for D351 phosphorylation at the cytoplasmic domain P. Thereby it is suggested that the accurate configuration and flexibility of L78 play important roles for the successive conformational changes accompanying the SERCA1a reaction cycle.]]> 2014/8/20 0:00:00 WANG Guo-Li,DAIHO Takashi,YAMASAKI Kazuo,DANKO Stefania,WANG Biao,SU Wen-Hui and SUZUKI Hiroshi 474true 2014/8/20 0:00:00 ZHAO Jun and WANG Jin-Hui 473true 2O2)．That result indicated that subcellular localization of Nmi can be affected by the oxidation environment of cells.]]> 2014/7/19 0:00:00 SHEN Juan and LIU Ying-Fang 472true 2014/7/19 0:00:00 LI Xin-Xin,WANG Ju-Ke and LIU Xin-Qi 471true P < 0.05). According to PPARδ gene, the designed PPARδ siRNA primer silenced the PPARδ expression and depressed the induction of GW501516 on PAI-1 expression. Then, the HUVECs were treated by SB431542 or SIS3, which is the TGFβ-Smad3 signaling pathway blocker, and found that PAI-1 expression of cells were down-regulated.At last, we found that SB431542, SIS3 and GW501516 inhibited the expressions of pSmad3 protein. These results suggested that TGF beta-Smad3 signaling pathway involved in the regulation of GW501516-induced PAI-1 expression in HUVECs.]]> 2014/7/19 0:00:00 MO Zhong-Cheng,LI Xia,ZHANG Da-Di,ZHU Ming-Yan,ZHANG Qing-Hai,ZENG Ying,WU Rong,LU Yan-Ju,CHEN Yi and YI Guang-Hui 470true WRKY42 gene was occurred at seedling stage and anther, while the expression of WRKY42 protein was detected in the leaf blade at all growth stages. In the process of Xa21- mediated resistance to bacterial blight, the induction of WRKY42 protein was apparent in the late stage after inoculation; further comparison among incompatible (R), compatible (S) and mock control (M) reactions revealed similar expression pattern between R and S interactions, and the abundance of WRKY42 in both R and S interactions was significantly greater than that in mock control reaction. This evidence suggested that WRKY42 protein may play a role in the interactions between rice and Xoo. Next, the WRKY42 protein was expressed in E. coli. The purified WRKY42 protein was used to investigate its binding capacity with cis-elements in the promoter region of downstream genes using microscale thermophoresis (MST) technique. It was found that the WRKY42 protein binds to W-box-containing oligonucleotide derived from the promoter region of PR1a and PR1b specifically, and their dissociation constant (Kd) were 73.3 μmol/L and 58.3 μmol/L, respectively. Taken together, the data provide direct evidences for the function of WRKY42 to regulate downstream pathogenesis-related genes, and further support that WRKY42 play a role in the process of rice resistance to Xoo. In the last, a working model of WRKY transcription factors in rice-Xoo interactions was proposed.]]> 2014/7/19 0:00:00 MIAO Liu-Yang,ZHOU Liang,YANG Shuo,LI Li-Yun,LI Xue-Jiao,FAN Wei,LAN Jin-Ping,SHI Jia-Nan,LIU Li-Juan and LIU Guo-Zhen 469true 2014/7/19 0:00:00 TAO Xian-Ming,WANG Peng-Fei,WANG Yang and LIANG Xing-Guo 468true Vibrio harveyi is a serious infection which brings a heavy damage to aquaculture. Accurate and rapid detection of the microorganism is the first and necessary step to control the disease. Aptamers have a good potential application in the detection and identification of microorganisms because of their strong affinity, high specificity and good stability. In the present paper, aptamers targeting to V. harveyi were selected by the technique of systematic evolution of ligands by exponential enrichment (SELEX), and their affinities and specificities to V. harveyi were also evaluated. After 15 rounds of selection, the affinity of the aptamers in the enriched pool increased to 58.95, which was 16.8 times of that (3.51) in the original pool. After cloning and sequencing, 52 supposed aptamers were obtained from the enriched pool. These aptamers were divided into 8 families based on the homology analysis. Over 50% of these aptamers in the first and the second families shared higher homology, suggesting the selection was convergent and efficient. Six high-frequency aptamers were also found to have significant affinities and specificities to V. harveyi (P < 0.01). Five of them (S1,S25, S26, S27, S35) were proved to have higher affinities, and their affinity constants (Kd) were as followed: (32.6±7.1), (45.3±10.1), (24.7±5.8), (34.8±5.6) and (12.9±4.0) nmol/L, respectively. The production mechanism and the application values of high-frequency aptamers were also discussed. The present paper was the first report of aptamers with high affinity and specificity to the target V. harveyi, which would lay the foundation for the detection of the pathogenic microorganism based on aptamers.]]> 2014/7/19 0:00:00 ZHENG Jiang,HAO Ju-Min,SONG Lin-Sheng and LIU Rui 467true 2014/6/19 0:00:00 LI Hong-Yan,ZHOU Zhi-Ming,PAN Qiang,YANG Dong-Mei,GU Jin-Fa,SHEN Jia-Ni,WANG Shi-Bing and ZHANG Kang-Jian 466true 2014/6/19 0:00:00 XING Yu-Hua,LIU Da-Tao,TAN Jun-Jie,HU Li-De,LIU Gang,FU Xue-Qi and CHEN Hui-Peng 465true in vivo function, we generated a transgenic mouse line overexpressing miR-30b under the control of the 5.5 kb promoter of α-myosin heavy chain (α-MHC). qRT-PCR results demonstrated that miR-30b was significantly increased in the heart tissues of miR-30b transgenic mice (P < 0.05). miR-30b transgenic mice did not exhibit significant heart/body weight and left ventricular(LV)/body weight changes and abnormal myocardium structure. At present, little is known about how miR-30b regulates myocardial infarction. We constructed I/R models by the coronary artery ligation method and sham-operated mice were used as controls. Biochemical detection results and TTC-Evans blue results showed that after ischemia-reperfusion, these transgenic mice had lower releases of LDH, CK and cTnⅠ(P < 0.05)and their hearts exhibited a smaller infarct size compared to those from control mice(P < 0.05). Echocardiographic results indicated that cardiac function of transgenic mice was markedly improved compared to that of control mice. In conclusion, miR-30b has protective effect upon ischemic-reperfusion injury. And it may provide a new therapeutic approach for preventing and treating myocardial infarction.]]> 2014/6/19 0:00:00 FAN Yuan-Yuan,LONG Bo,LIU Fang,ZHOU Lu-Yu,WANG Kun and LI Pei-Feng 464true IC50 values of indometacin, aspirin and acetaminophen were 0.254, 0.564 and 0.656 mmol/L, respectively; and their average inhibitory constants were 0.154, 1.38, and 2.98 mmol/L, respectively. Competitive inhibitory type was observed for all the three drugs by plotting the Lineweaver-Burk curves. According to the kinetics of the hydrolysis, IC50 values, inhibitory constants, and Michaelis constants, the inhibitory abilities of the three drugs were ranked as: indometacin>aspirin>acetaminophen. This chemiluminescent method is especially valuable for evaluation of those drugs which the UV method cannot work for.]]> 2014/6/19 0:00:00 MU Xiao-Jing,GONG Yu-Hua,CHEN Zhi-Tao,LIU Min,BI Wen-Jie and XU Hong 463true 2014/6/19 0:00:00 LIU Jing,ZHU Hong-Tao,FENG Hong-Li,GONG Min-Qing and ZHU Ping 462true 2014/6/19 0:00:00 XU Xian-Jin,SU Ji-Guo,CHEN Wei-Zu,WANG Cun-Xin,CANNISTRARO Salvatore and BIZZARRI Anna-Rita 461true 2014/6/19 0:00:00 TAN Heng and WANG Yi 460true Drosophila innate immunity, Drosophila mutants, E(spl) and Su(H), which are located in the downstream of Notch signaling pathway, were utilized to analyze the survival rate, in vivo phagocytosis and production of antimicrobial peptides after pathogen infections. In addition, the number of circulating plasmatocytes in third instar larvae of mutant and wild type flies was also measured. The results indicate that the E(spl) mutant showed lower survival rate, phagocytosis and the expression of AMP genes after pathogen infections. Furthermore, an abnormal increase in the number of plasmatocytes in E(spl) mutant larvae was also observed. Compared with E(spl) mutant, the mutant of Su(H) only showed sensitive to fungi, however, phagocytic ability of hemocytes was normal in fungi infection. At the same time, the expression of antimicrobial peptides was decreased in Su(H) mutant. These results show that the Notch pathway not only affects the growth and development of individuals, but also plays an important role in regulating innate immunity of Drosophila.]]> 2014/5/22 0:00:00 WANG Xiao-Chun and JIN Li-Hua 459true 2014/5/22 0:00:00 DONG Chuan,CAO Zan-Xia,ZHAO Li-Ling,SUO Zhen-Peng and WANG Ji-Hua 458true 2014/5/22 0:00:00 LI Shan,LIU Bin,LI Chun-Hua,TAN Jian-Jun,ZHANG Xiao-Yi and WANG Cun-Xin 457true 2014/5/22 0:00:00 YANG Zhi-Lai and WANG Jin-Hui 456true 2S) against the neurotoxicity and oxidative stress of formaldehyde (FA). Paraoxonase-1 (PON-1) is a pivotal endogenous antioxidant. The aim of this present study is to investigate whether PON-1 mediates the protection of H2S against FA-induced neurotoxicity. In the present work, PC12 cells treated with FA were established to the model of FA-induced neurotoxicity. Treatment of PC12 cells with NaHS (a donor of H2S) not only upregulates the activity of PON-1，but also significantly restores FA-induced downregulation of PON-1 activity and expression. A selective inhibitor of PON-1, 2-hydroxyquinoline (2-HQ), markedly attenuated H2S-induced neuroprotection against FA-induced cytotoxicity, apoptosis, and accumulation of intracellular reactive oxygen species (ROS) in PC12 cells. Furthermore, 2-HQ blocks H2S to reverse FA-caused activation of caspase-3 and downregulation of bcl-2 expression in PC12 cells. These results indicate that H2S protects PC12 cells against FA-induced neurotoxicity in a PON-1-dependant manner. Our findings suggest a promising role of PON-1 as a novel therapeutic target for neuronal damage after FA exposure.]]> 2014/5/22 0:00:00 ZHANG Ping,LI Xiang,REN Yan-Kai,WANG Chun-Yan,ZOU Wei and TANG Xiao-Qing 455true IT/IC), which would cancel out the influence of the stability of QDs and antibody. The immunoreaction time and the sample volume are optimized to get a higher sensitivity. Under optimal conditions, the QD-based lateral flow immunoassay system has a wide dynamic range and is capable of detecting a minimum 0.85 IU/L HCG standard analyte in 15 min with 50 μl sample volume. The novel QD-based lateral flow immunoassay biosensor offers a rapid, sensitive and quantitative tool for point-of-care testing of the level of serum HCG and shows great promise in other protein markers.]]> 2014/5/22 0:00:00 YANG Qiu-Hua,WANG Xin,YANG Jiu-Min,LI Xue,SONG Tao,GONG Xiao-Qun,ZHANG Bing-Bo and CHANG Jin 454true in vitro through co-immunoprecipiation in combination with Western blotting and immunostaining. Knock down of endogenous CLU expression in osteosarcoma cells significantly inhibit cell proliferation and cell invasion ability in vitro. This preliminary study on the molecular mechanisms that how IEX-1 promote the occurrence and development of osteosarcoma provides novel clues for early-stage diagnosis and target gene therapy of this malignant bone tumor.]]> 2014/5/22 0:00:00 LIU Li-Hong,LI Hui,LI Jie,OUYANG Yu-Rong,GUO Hon-Bin,CUI Yun-He,LIU Ying-Qi and XIAO Tao 453true 2014/4/23 0:00:00 YI Mei,LI Ji,SU Juan,ZHANG Jiang-Lin,CHEN Xiang,DU Qian-Jun,XIANG Bo,XIONG Wei,LI Xiao-Ling and XIE Hong-Fu 452true via the tail vein into HBV transgenic mice or hydrodynamically injected mice. In the HBV transgenic mice, with the treatment of X-519, HBV pregenomic RNA in the liver decreased by 81%. Cationic liposome further increased the inhibition effectory to 91%. But no significant differences were observed for HBV antigens and HBV DNA copy number in the sera. In acute HBV infection mouse model by hydrodynamic injection, ELISA and quantitative real-time PCR analysis showed that X-519 significantly repressed HBV replication, as measured by HBV pregenomic RNA, antigens expression, and presence of HBV DNA in the sera. Taken together, the synthesized antisense RNA oligonucleotide X-519 repressed HBV replication and antigens expression in vitro and in vivo.]]> 2014/4/22 0:00:00 ZAN Yan-Lu and TIAN Bo 451true 2014/4/22 0:00:00 WANG Hai-Long,DENG Yu-Lin,ZHANG Ze-Bo,WANG Chen,YANG Zu-Ye,LI Fu-Tao and MA-Hong 450true 2014/4/22 0:00:00 CAO Jia-Yue,FENG Zhou-Yan,ZHENG Xiao-Jing and CHEN Bai-Lu 449true 2014/4/22 0:00:00 CHEN Xiao-Bin,ZHU Wei-Bin,RUAN Lin-Hao and TANG Jie 448true in vivo adult brain plasmid electroporation to label mature GCs in the OB directly were adopted. Here, the spines in EPL were defined as distal spines and the spines in GCL as proximal spines. It was revealed that the density of spines on granule cells decreased significantly in EPL (distal spines) of CNGA2 KO mice, but did not change significantly in GCL (proximal spines), as same as the result of optical density analysis in the VGluT1 immunolabeling. These data suggest that the structural plasticity of the distal dendrodendritic synapses, rather than the proximate axon-dendritic synapses on granular cells of the OB, are significantly affected by the peripheral olfactory inputs.]]> 2014/2/21 0:00:00 RAO Xiao-Ping,XU Zhi-Xiang,WANG Li and XU Fu-Qiang 447true 2014/2/21 0:00:00 DENG Chun-Shan and WANG Yi 446true in vitro. The results showed that calcium signaling responded to mechanical stimulus and propagated to neighboring cells in isolated BV-2 microglial cells, suggesting that the paracrine pathway mediated intercellular calcium waves. Further data indicated that gap junction was involved in mechanically-induced intercellular calcium waves when the cells had physical contact with each other. More importantly, tunneling nanotubes (TNTs)-like structure between BV-2 microglial cells was found in vitro and mediated the propagation of intercellular calcium wave. Taken together, our studies demonstrated that three distinct pathways, including paracrine, gap junction and TNTs-like structures, could mediate the intercellular calcium waves in BV-2 microglial cells.]]> 2014/2/21 0:00:00 WU Xian,PAN Lei-Ting,LIU Ying,XING Meng-Xi,ZHANG Xin Zheng and XU Jing-Jun 445true 18-labeled MVs were injected intravenously into BALB/c mice; the frozen section of liver was observed through fluorescent microscopy. Finally, miR-122 targeting AldoA in the metabolism of ob/ob mice was confirmed by qRT-PCR and Western blotting. AldoA mainly catalysed the transformation between dihydroxyacetone phosphate, glyceraldehyde-3- phosphate and fructose 1, 6 - bisphosphate in glycolytic pathway. MiR-122 may play an important role in the pathologenesis of ob/ob mice through AldoA pathway.]]> 2014/2/21 0:00:00 FANG Zhi-Juan,LI Peng,DIAO Wen-Li,JIANG Ting,ZHANG Chen-Yu and XIANG Yang 444true Saccharomyces cerevisiae) protein kinase Sch9 is homologous to the mammalian kinase S6K1. S6K1 is a substrate of mammalian target of rapamycin (mTOR) and phosphatidylinositol-3 kinase (PI3K) and relates to many diseases, including obesity, diabetes and cancer. Both Sch9 and S6K1 are important to the regulation of cell growth in response to different nutrient and stress factors. The residue T570 is a conserved phosphorylation site in the activation loop of Sch9, also called PDK1 site. Whereas another conserved phosphorylation site, T737, in the hydrophobic motif of C terminus is called PDK2 site. The phosphorylation of these two sites are important to Sch9 kinase activity. Upstream kinases Pkh1/2 phosphorylate the PDK1 site, while the Target of Rapamycin Complex 1 (TORC1) phosphorylates the PDK2 site. To better understand the intracellular function of protein kinase Sch9, it is important to elucidate the dynamics and regulation of the phosphorylation of PDK1 and PDK2 sites in Sch9 under different environmental condition. Using antibody that is specific for T570 site phosphorylated Sch9 or T737 site phosphorylated Sch9, we studied the regulation of the phosphorylation of PDK1 and PDK2 sites in Sch9 under different environmental factors and during chronological aging. Our results demonstrate the regulatory model of the phosphorylation of PDK1 and PDK2 sites in Sch9 during nutrient sensing, stress response, calorie restriction and chronological aging. The results also suggest a novel mechanism by which calorie restriction extends chronological lifespan that involves the regulation of the phosphorylation of Sch9 PDK1 site.]]> 2014/2/21 0:00:00 LIU Jun,LV Lei,QIE Bei-Bei and LIU Ke 443true 2 and (102.73±13.81) μg/cm2 respectively, the HA-GMS nanoemulsion significantly increased the efficiency of MD-CPT through skin, which may provide a good drug delivery approach.]]> 2014/2/21 0:00:00 GAO Yuan-Yuan,KONG Ming,CHENG Xiao-Jie,WANG Zhi-Guo and CHEN Xi-Guang 442true in vitro using IL-15 followed by transferring them into mice with B16-OVA melanoma. We found that TCM conferred a prolonged suppression of tumor growth whereas effector T cell (TEFF) cultured by IL-2 showed shorter suppression for tumor growth, which was correlated with sustained high number of tumor-specific CD8 T cells in spleen from IL-15 group. Interestingly, the expression of MHC-Ⅰ in the tumor from IL-15 group was upregulated which suggested that the cross presentation in IL-15 group was efficient. These data collectively suggested that IL-15 treated CD8 T cells elicit more effective anti-tumor response than IL-2 treated CD8 T cells owing to sustained tumor-specific CD8 T cells in spleen. Furthermore, expression of MHC-Ⅰwas upregulated in tumor in response to IL-15 mediated adoptive T cell transfer, suggesting that antigen presenting cells (APCs) were involved in adoptive T cell transfer. Our research may have implications for developing more effective tumor immunotherapy.]]> 2014/12/19 0:00:00 JU Jian-qi,JIA Ming-Ming,CONG Yan-Ni and WANG Sheng-Dian 441true P < 0.01) ，and there was a very significant difference in BA between the S neurons of harmonic and non-harmonic neurons (95.3 ± 14.0 vs 109.7 ± 7.9 dB SPL, P < 0.01). These results suggested that there is a clear division of labour in IC neurons processing and analyzing echo information during the DSC. S neurons which concentrated in the harmonics can effectively encode the echo information and avoid disturbances of other clutter information by increasing the recovery rate of responses to echo under specific frequency shifts of DSC. And the characteristics of firing patterns and intensity-latency functions of S neurons of harmonic neurons were different than other neurons，which met the need in precise acoustic imaging at complex environments.]]> 2014/12/19 0:00:00 LI Lin,FU Zi-Ying,WANG Li-Ping,WEI Chen-Xue,ZOU Li-Fang,CHEN Qi-Cai and TANG Jia 440true 2014/12/19 0:00:00 PAN Yao-Qian,PAN Bo,LIU Xing-You,LI Rui-Zhen and YUE Jiun-Ming 439true 1/2 respectively compared with intravenous and intramuscular injection; moreover, the curve of transdermal delivery was smooth continuous. In vivo imaging system intuitively reflected the distribution of transdermal drug in mice and the drug content of various organs/tissues. The rest of MD-CPT and HANs entered into the blood circulation, eventually excreted through metabolism without body burden. The longer drug residence at topical area with treatment could provide a continuous and controllable, extended efficacy, which was beneficial to superficial lesions therapy.]]> 2014/12/19 0:00:00 GAO Yuan-Yuan,WANG Zhi-Guo,JIANG Chang-Qing,KONG Ming,CHENG Xiao-Jie,WANG Juan,BAO Zi-Xian,SUN Guo-Hui,HUANG Yu-Hua,LIN Cheng-Bing,YI Hong-Jun and CHEN Xi-Guang 438true 2014/11/21 0:00:00 ZHAO Ping,ZHOU Qi,BI Long-Jun,SHEN Jin-Hua,YANG Xin-Zhou and LIU Qing-Hua 437true 2014/11/21 0:00:00 DU Jing,ZU Yan,XU YUE and YANG Chun 436true IGFBP2 is the only known gene located within this QTL region. We further found that the 1196C>A, a single nucleotide polymorphism (SNP) in 3'UTR of Chicken IGFBP2 gene, is significantly associated with chicken abdominal fat weight and percentage of abdominal fat. Bioinformatics analysis suggests that this SNP is located at a potential binding site of gga-miR-456-3p. We hypothesized that IGFBP2 may be a target gene of gga-miR-456-3p, and this SNP may affect the gga-miR-456-3p-mediated downregulation of IGFBP2. To test and decipher this hypothesis, in the present study we constructed the 3′UTR reporter vectors for the two individual alleles of the IGFBP2 SNP (1196C>A), and compared the reporter activity between the two individual alleles in both DF1 cells and chicken preadipocytes. Using the miRNA mimics and inhibitor of gga-miR-456-3p, we further assessed the effects of gga-miR-456-3p on the reporter activity of the two individual alleles of the IGFBP2 SNP and the endogenous IGFBP2 expression at mRNA and protein levels in DF1 cells. The reporter assay showed that in both DF1 cells and chicken preadipocytes, allele A had higher reporter activities at protein and mRNA levels than allele C, indicating that this SNP is a functional SNP. Further studies demonstrated that in DF1 cells, gga-miR-456-3p mimcs and inhibitor had effect on allele C reporter activity, but not on allele A reporter activity. Quantitative real-time RT-PCR and Western blot analyses showed that gga-miR-456-3p mimcs and inhibitor regulated the endogenous expression of chicken IGFBP2 gene at mRNA and protein levels. Taken together, our results demonstrated that IGFBP2 gene is a target gene of gga-miR-456-3p, and the SNP 1196C> A is a functional SNP. Our findings are of great significance to the marker-assisted selection for lean chicken and clarification of gene function and regulation of chicken IGFBP2 gene in chicken adipose development.]]> 2014/11/21 0:00:00 YU Ying-Ying,QIAO Shu-Pei,SUN Ying-Ning,SONG He,ZHANG Xiao-Fei,YAN Xiao-Hong,LI Hui and WANG Ning 435true Sinorhizobium meliloti adaptation to external environment stress and symbiotic nodulation. The symbiotic plasmid A of S. meliloti encodes a DesA homologue, which has 44% identity to Bacillus subtilis DesA. However, whether S. meliloti DesA is able to play functions in fatty acid synthesis was not determined. We report that though SmdesA gene could not restore the growth of E. coli fabA temperature sensitive mutant CY57 under non-permissive conditions, argentation thin layer chromatography assay show that CY57 carrying desA encoded plasmid could produce palmitoleic acid and octadecadienoic acid. This indicate that SmdesA encodes acyl-lipid desaturase. In addition, deletion of SmdesA gene have led to a little effects on the fatty acid composition of S. meliloti, but show a significant impact on growth rate in low temperature and high salt conditions, and the ability of alfalfa symbiotic nodulation. DesA is not the key enzyme of unsaturated fatty acid synthesis of S. meliloti, but it plays important biological roles in S. meliloti dealing with environment stress and nodular process.]]> 2015/8/18 0:00:00 MA Jin-Cheng, ZHOU Jun-Chao, WU Chu-Yun, HU Zhe and WANG Hai-Hong 434true 2015/8/18 0:00:00 GUO Yu, SU Song-Kun, CHEN Sheng-Lu, ZHANG Shao-Wu and CHEN Run-Sheng 433true 2015/8/18 0:00:00 DU Juan, WU Xia, QI Hong-Xin, SONG Wei-Tao, WANG Zhi-Dan, RAO Ling-Yue, ZHANG San-Jun, CHEN Shu-De and XIA Ruo-Hong 432true 2015/7/20 0:00:00 MO Wei-Chuan, FU Jing-Peng, DING Hai-Min, LIU Ying, HUA Qian and HE Rong-Qiao 431true ENV), polar of FFR's instantaneous phase difference, vector of FFR's grand averaged phase difference, and spectrum signal-to-noise ratio of temporal-fine-structure-related frequency-following response (FFRTFS). Results show that compared to non-musicians, listeners with musical training were both more accurate at reporting the target digit and had a shorter reaction time, especially for attend-pitch trials. For both groups, noise had little effect on the neural encoding of the fundamental frequency (F0) but significantly degraded the neural encoding of harmonics. Compared to non-musicians, musicians showed enhanced phase locking of F0 as well as more robust phase locking to stimulus harmonics in the noise condition. Neural spectrum SNRs of FFRTFS to harmonics were correlated with listeners' behavioral correction rate on the pitch-based auditory attention, for which robustness of musicians' FFR out performed the non-musicians'. These findings suggest musician's better pitch perception in auditory selective attention is relevant to their enhanced neural cognition. Musical training is assumed to be able to enhance the FFRENV phase locking to the F0, the robustness and continuous phase locking of FFRTFS to the harmonics, and spectrum SNR FFRTFS to the harmonics. In conclusion, musical training has a significant effect on listeners' plasticity in auditory selective attention.]]> 2015/7/20 0:00:00 ZHU Li, ZHENG Fen, DENG Juan, YAN Zheng and XU Xiao-Ling 430true 2015/7/20 0:00:00 LEI Zhuo-Fan and WANG Jin-Hui 429true 2015/6/24 0:00:00 LI Xue-Tian, SUN Yu, DU Ya-Rong, WANG Xi-Long, ZHANG Pei-Xin, ZHONG Yao-Gang, MA Tian-Ran, WANG Xiu-Rong and LI Zheng 428true n. Our results showed that the levels of 10 species of LysoPCs were higher in LM-MCF-7 cells than that in MCF-7 cells. Meanwhile, six species of PCs that may produce arachidonic acid (AA) and associated LysoPCs were lower in LM-MCF-7 cells than that in MCF-7 cells. Furthermore, we identified that 4 out of 6 species of PCs were responsible for the generation of AA mediated by cPLA2 in LM-MCF-7 cells. The increased ratio of LysoPCs to PCs corresponds to the activation level of cPLA2. Thus, we conclude that 4 identified species of PCs and 4 corresponding species of LysoPCs mediated by cPLA2, as well as the up-regulation and activation of cPLA2, may involve in the metastasis of breast cancer cells.]]> 2015/6/24 0:00:00 YOU Jia-Cong, YANG Jie, FANG Run-Ping, HU Nan, ZHANG Xiao-Dong, ZHANG Wei-Ying and YE Li-Hong 427true 2015/6/24 0:00:00 LIN Lin, SONG Xian-Qiang, YE Sheng and ZHANG Rong-Guang 426true Salmonella paratyphi A (S. paratyphi A) mutants resistant to phage were analyzed in this study. Phages were dropped on bacteria line on LB plate containing Rif+(200 mg/L)to confirm phage resistance. Six mutants were cultured in 96 well plate with three replications to monitor biofilm formation and morphology. Site-directed mutagenesis and complementation test were used to confirm whether phage resistance was caused by gene mutant. Results of this study showed that all six mutants were resistant to phage, because those could growth with dropped 2.4×109 phages and wild bacteria could not with dropped 1.2×106 phages. Among six mutants, BF formation of mutant of σ-54-dependent translation regulator was significantly enhanced contrast with wild bacteria (P < 0.05, 1.1±0.2 vs 0.5±0.1). Bacteria aggregated to large irregular mass. Site-directed mutation of σ-54-dependent translation regulator with homologous recombination led to be resistant to phages. This resistance could be recovered by supplemented with plasmid expressed σ-54-dependent translation regulator. All those showed that σ-54-dependent translation of the regulator was associated with anti-phage and BF formation.]]> 2015/6/24 0:00:00 MAO Pu-Jia, FENG Meng-Die, HONG Yu, XU Ze-Yang, ZHAO Ji-Hua, YANG Hong-Wen, SONG Wu-Zhan, HUANG Fen, JING Shen-Rong and ZENG Wei-Kun 425true 2015/5/22 0:00:00 TIAN Xiao-Fei, MA Wei-Juan, FANG Guang-Guang, XIAO Tian-Xia, CHEN Jie and ZHANG Jian 424true 2015/5/22 0:00:00 LU Wei, YANG Gang-Long, XUE Peng, LIU Chang-Mei and GUAN Feng 423true 2015/5/22 0:00:00 WANG Cheng, HUANG Si-Si, LU Li, GU Yong-Qing and ZHANG Ling-Qiang 422true R=0.093) and A1C levels (R=0.1246). Breath Acetone concentration was significantly higher in male subjects (1.75×10-6) than female subjects (1.15×10-6), and male subjects' breath acetone was clearly increased with decreasing age (R=-0.154). Besides, breath acetone concentration was significantly correlated inversely with BMI (R=-0.2) in male subjects, and significantly higher in subjects with BMI under 25 (1.75×10-6) than obese subjects with BMI over 25 (1.25×10-6). Breath acetone was correlated positively with years of diabetes in female subjects (R=0.17,). Whereas, negatively correlated in male subjects (R=-0.14). Furtherrnore, there were variations in relation to breath samples' collection conditions in both male and female subjects. Multi-variation analysis showed that gender (β=0.374)、A1C (β=0.083)、BG (β=0.002)、breath samples' collection conditions (β=-0.289)、BMI (β=-0.046) and age (β=-0.009) affect breath acetone concentrations. In conclusion, breath acetone concentration was correlated with gender, breath samples' collection conditions, BMI, age, A1C and BG.]]> 2015/5/22 0:00:00 CHEN Zhu-Ying, SUN Mei-Xiu, GONG Zhi-Yong, YUAN Yuan, ZHAO Xiao-Meng, LIU Wei-Chao, JIANG Chen-Yu, WANG Zhen-Nan and LI Ying-Xin 421true 2015/4/24 0:00:00 ZHANG Qing, YANG Hui, LI Jia, XIE Xin and LI Jing 420true 2015/4/24 0:00:00 WANG Hao, SHE Qiang, GAO Ling-Zhi, ZHA Cheng-Qin, DU Jian-Lin and JING Xiao-Dong 419true 2015/4/24 0:00:00 XUE Peng, ZHOU Yue, DING Xiang, WANG Jun, XIE Zhen-Sheng and YANG Fu-Quan 418true 2015/4/24 0:00:00 YU Jing, MA Zhen-Ling, NIU Ya-Nan, ZHANG Bao-Shan, Lucas S. Broster and LI Juan 417true Acinetobacter baumannii is a new threat in intensive care units (ICUs) for its multiresistance to antibiotics, but little is known about this bacterium. Nucleoside diphosphate kinase (NDK) is an evolutionarily conserved enzyme that catalyzes phosphoryl transformation between nucleosides. In our study, the crystal structure of wild type Acinetobacter baumannii NDK along with its mutant generated through truncation of the C-terminal arginine-threonine-arginine (RTR) residues, were solved. In comparison with Myxococcus xanthus NDK structure, we speculated that Acinetobacter baumannii NDK shared a similar catalytic mechanism with Myxococcus xanthus. Activity assay and CD spectra analysis revealed that E28A mutant might interrupt the secondary structure of the protein leading to declined enzymatic activity. Truncation of the C-terminal RTR residues would lead to the instability of the tertiary structure resulting in reduced kinase activity. Lys33 was a key residue for maintaining dimer interaction when RTR residues were truncated but was not sufficient to keep efficient enzymatic reaction. The structural data can provide a potential target to develop novel therapeutic approaches to overcome multiresistance of the bacterium against antibiotics.]]> 2015/3/24 0:00:00 HU Ying-Song, FENG Feng and LIU Ying-Fang 416true nptⅡ gene, encoding neomycin phosphotransferase Ⅱ, is one of the most commonly used selection marker in transgenic research. In this study, recombinant NPTⅡ protein was expressed in E. coli and monoclonal antibodies were generated. The inheritance, temp-spatial profiling, abundance and subcellular localization properties were investigated using established Western blot (WB) analysis. The results showed that NPTⅡ protein in 0.25% of a single rice grain (about 0.025mg) was detectable, the expression of NPTⅡ protein is consistent with dominant inheritance law, positive seeds and homozygous lines can be identified in transgenic T1 generation, furthermore, the abundance of NPTⅡ protein can provide clues for the identification of homozygous individual in T1 seedlings. Quantitative analyses shows that NPTⅡ protein accounts for about 0.08‰ of total protein in the leaves at seedling stage. The expression profile of NPTⅡ protein in transgenic rice shows that the abundance of NPTⅡ driven by CaMV-35S promoter in the leaves and roots at seedling and adult stage are higher than that at tillering and booting stage. NPTⅡ was detectable at different stages in rice root, stem, leaf, panicle, flower and seed, however, the abundance of NPTⅡ in tillers, node, panicle axis and anther tissues is lower than others. In addition, the NPTⅡ protein is mainly localized in the soluble portion of cytoplasm. Taken together, an applicable immunoblot method was established and the expression properties of NPTⅡ protein in transgenic rice were demonstrated.]]> 2015/3/24 0:00:00 LAN Jin-Ping, WU Peng-Cheng, GUO Mei-Cen, SHI Jia-Nan, WEI Han-Fu, RONG Rui-Juan, HAO Yu-Jie, YANG Shuo, BAI Ying-Ying, LI Li-Yun, WU Lin, LIU Si-Qi, YIN Chang-Cheng and LIU Guo-Zhen 415true Uchl1 in substantia nigra region of MPTP-treated mice were significantly higher than that in the saline controls, with significant decreased expression of mRNA and protein. The methylation levels of promoter region of Arih2 in substantia nigra region of MPTP-treated mice were significantly decreased than that in the saline controls, with significant increased expression of mRNA and protein. These results suggested that DNA methylation were altered in substantia nigra region of MPTP-induced PD model. DNA methylation may play important role in the pathogenesis of PD induced by environmental factors such as MPTP.]]> 2015/3/24 0:00:00 HU Ya-Cen, XU Qian, GUO Ji-Feng, AI San-Xi, SONG Cheng-Yuan, SUN Qi-Ying, WENG Ling, ZHOU Lin, JIANG Hong, SHEN Lu, YAN Xin-Xiang and TANG Bei-Sha 414true 2015/2/6 0:00:00 LI Yang, YAN Cheng-Hui, TIAN Xiao-Xiang, PENG Cheng-Fei and HAN Ya-Ling 413true IC50 were significantly decreased (P < 0.05). The IC50 values of the combination for 24, 48 and 72 h were 2.83, 3.15, 2.19 fold and 2.66, 3.11, 2.45 fold respectively compared to BGC-823 cells treated with CPT-11 and NCTD alone. The results indicated synergistic effect. The sequence of CPT-11 followed by NCTD showed a stronger inhibition than the sequence of NCTD followed by CPT-11 (P < 0.05), and it was superior to co-administration (P < 0.05). For 24 h, CPT-11 60 μmol/L triggered both S and G2-M phase arrest (P < 0.01) in the BGC-823 cells, and NCTD 60 μmol/L induced cell cycle arrest at G2-M phases and apoptosis (P < 0.05) in the BGC-823 cells. The combination of CPT-11 and NCTD (60∶60) μmol/L increased the G2-M phase arrest. Compared with the sequence of NCTD 6 h first followed by CPT-11 and co-administration, the sequence of CPT-11 6 h first followed by NCTD increased the S phase arrest (P < 0.05) and apoptosis in the BGC-823 cells. CPT-11 30，60 μmol/L up-regulated the expression of Pdcd4 (P < 0.05) and down-regulated the expression of p53(P < 0.05) after 12 h; NCTD 30，60 μmol/L down-regulated the expression of Pdcd4 (P < 0.05) and up-regulated the expression of p53(P < 0.05) after 12 h; The combination (30∶30, 60∶60) μmol/L up-regulate the expression of Pdcd4 and p53(P < 0.05). The combination of CPT-11 and NCTD has a synergistic effect mainly due to the fact that it can induce the G2-M phase arrest, and up-regulate the expression of Pdcd4 and p53 as the tumor suppressor. The combination of CPT-11 and NCTD with a sequential schedule has some impacts on the growth of BGC-823 cells. The sequence of CPT-11 followed by NCTD shows a stronger inhibition than the sequence of NCTD followed by CPT-11 and co-administration, and it may be related to the increase of S phase arrest and apoptosis. Pdcd4 and p53 protein may have a negative regulation in BGC-823 cells.]]> 2015/2/6 0:00:00 LI Zi-Mu and SUN Zhen-Xiao 412true 2015/2/6 0:00:00 CHEN Ke, YIN Jiao-Jiao and SONG Xue-Mei 411true 3) of Epstein-Barr virus encoded latent membrane protein 1(LMP1) in nasopharyngeal carcinoma stem cell SP18 cells, the SP18 cell of stable expressed LMP1 and deletion mutant type LMP1 (LMP1△232-351) were established (SP18-LMP1 and SP18-LMP1△252-351). The effect of LMP1-CTAR3 deletion mutant for cellular proliferation, migration and invasion were observed in SP18 cells. The differential expression genes between SP18-LMP1 and SP18-LMP1△252-351 cells were analyzed by cDNA chips, and the expression levels of partial identified genes were verified by fluorescent Real-time quantitative RT-PCR. The relation of differential expression genes were analyzed by bioinformatics. The results showed: (1) The ability of LMP1△232-351 promoting SP18 cell proliferation, migration and invasion was obviously decreased to compare with wild type LMP1(n=3, P < 0.05). (2) 18 genes, 13 up- and 5 down-regulated ones, of LMP1-CTAR3 mediated regulation with migration and invasion were identified from in SP18 cell lines. The differential expression of partial identified genes was similar with cDNA chips separated ones and confirmed by fluorescent Real-time quantitative RT-PCR. (3) 13 differential expression genes can be cross-talk, and among FN1, MMP14, ITGA2, THBS1, IL1B and IL6 genes were frequently correlation. These results suggested that LMP1-CTAR3 probably regulates the expression of FN1, MMP14, ITGA2, THBS1, IL1B and IL6 genes to induce and promote migration and invasion of nasopharyngeal carcinoma stem cell SP18 cell.]]> 2015/1/19 0:00:00 XIAO Juan, ZHANG Zhi-Wei, WU Shi-Hua, LIU Zhong-Yuan, TANG Wei, ZHOU Qiang, LEI Ming-Sheng and YANG Dai-Shui 410true 2015/1/19 0:00:00 LI Hui-Min, SHI Yu, YANG Zhi-Gang and JIANG Shao-Ping 409true 2015/12/18 0:00:00 LENG Fang-Wei, XIE Can and WANG Da-Cheng 408true 2015/12/18 0:00:00 WANG Juan, SHU Mao, LIN Yong, HU Yong, HAN Ying-Zi and LIN Zhi-Hua 407true 2015/12/18 0:00:00 HAN Jun-Ping, SUN Jing, ZHUANG Bin, LIU Peng, ZHAO Xing-Chun, LI Wan-Shui, JI An-Quan, YE Jian, LIU Yao and LI Cai-Xia 406true 2015/12/18 0:00:00 NANG Yi, MEI Han-Xue, ZHAO Yan, HOU Bao-Yan, ZHAO Zhi-Yuan and FAN Gou-Liang 405true 2015/10/20 0:00:00 ZOU Fei, DIAO Zhi-Qing, ZHANG Hong-Chao, XU Shi-Meng, LIU Ping-Sheng, LIANG Bin and WEI Xuan 404true Mycobacterium smegmatis using methods of tandem affinity purification and mass spectrometry and used Far-western and GST pull-down analysis to validate the interactions between MutM and DEAD-box rna helicase, RpsC, and UvrA. Results demonstrated that tandem affinity purification is a suitable method for identifying MutM interacting proteins and provided insights into the mechanism by which MutM is involved in DNA damage repair.]]> 2015/10/20 0:00:00 FAN Shang-Hua, ZHOU Ying, ZHANG Hong-Tai, Joy Fleming, YU Zi-Niu, ZHANG Xian-En and BI Li-Jun 403true 2015/10/20 0:00:00 LIAO Xiang, CHEN Fu-Yong, TAO Wei, LIU Ying, ZHANG Han, LI Yong-Jie and ZANG Yu-Feng 402true Euglena gracilis were investigated during the SIMBOX mission on board the Chinese Shenzhou-8 spacecraft. We found that microgravity decreased Fv/Fm, while chlorophyll a and carotenoid contents were increased. The fluorescence yields were clearly reduced by microgravity, but the shape of fluorescence transients (O-J-I-P) was not changed. The quantum yield of primary photochemistry (φPo), the quantum yield of electron transport (φEo) and the performance index of PSⅡ(PIABS and PICS) in microgravity group were decreased, while ABS/RC and DIo/RC were increased. 77K fluorescence emission spectra indicated that microgravity altered energy distribution between PSⅡ and PSⅠ and there are redshifts under microgravity. These results suggested that microgravity may impair photosynthesis by inhibition of acceptor sides of PSⅡ in electron transfer pathway and alter PSⅠ structure to cause a reduction of energy transfer to PSⅠ in Euglena gracilis.]]> 2016/9/18 0:00:00 LI Xiao-Yan, LI Gen-Bao, LI Dun-Hai, HAO Zong-Jie, WANG Gao-Hong, RICHTER R. Peter, SCHUSTER Martin, LEBERT Michael and LIU Yong-Ding 401true 2016/9/18 0:00:00 WANG Ling-Ling, HUANG Ya-Qi, ZHANG Xue-Fang, QU Jian-Bo and LIU Jian-Guo 400true P21, CD44 and BMI-1 upon treatments. MTT test demonstrated that VPA significantly (P < 0.05) increased toxicity of DDP, which was confirmed by DNA laddering, flow cytometry analysis and significant (P < 0.05) overexpression of P21. Moreover, real time RT-PCR results indicated significant (P < 0.05) down regulation of CD44 and BMI-1 after VPA administration. Present attempt provided evidence, for the first time, that VPA not only improved responsiveness of esophageal stem-like cancer cells to DDP, also negatively regulated cancer stem cells markers in these cells.]]> 2016/9/18 0:00:00 Saboor Maleki,Fatemeh B. Rassouli,MAryam M Matin and Mousavi 399true 2016/8/5 0:00:00 YAN Zhang-Ming, WANG Yao, LIU Ke, XIANG Shu-Nian and SUN Zhi-Rong 398true PstⅠ was able to cleave the pBR322 DNA Ⅳ molecules, indicative of the existence of an intact restriction site in the denatured DNA. AFM images revealed that PstⅠ-treated DNA Ⅳ molecules contained an individual node. The contour length of the condensed molecules was decreased by about 11% compared to DNA Ⅳ. Intriguingly, the Escherichia coli topoisomerase Ⅳ, a type Ⅱ topoisomerase, also introduced nodes into DNA Ⅳ while keeping the molecules in a closed form. The two types of DNA were similar to each other in terms of the contour length and the size of the node based on the AFM analysis. These findings suggest that specific base-paired regions exist in DNA Ⅳ, and that cleavages at these regions may lead to DNA node.]]> 2016/8/5 0:00:00 TIAN Lei, SU Xiao-Yun, ZHANG Zhen-Feng and HUANG Xi-Tai 397true i.e., the Schaffer collaterals) of the hippocampal CA1 region of anesthetized rats. Multi-channel spike signals were recorded in the downstream area of HFS by a microelectrode array. The features of the spike waveforms, such as amplitude and half-height width, were extracted from the sorted unit spikes of interneurons activated by HFS through single synaptic transmissions. The results show that the spike amplitudes decreased and the half-height widths increased significantly during HFS. The percent ratios of the waveform features, compared to their baseline values, show that the amplitude of falling-phase and the amplitude of rising-phase decreased by ～20% and ～40%, respectively, while the half-height widths increased by more than 10%. In addition, the decrease of amplitudes (up to 50%) and the increase of half-height widths (up to 20%) were enhanced during synchronized firings of a large population of neurons or during the O-HFS trains of 100 Hz, a stimulation frequency causing greater excitation to neurons than 200 Hz. Presumably, the excitation of O-HFS could elevate the membrane potentials of downstream neurons and change the kinetics of the ionic channels of the membrane, thereby result in the waveform changes of action potentials. The results support the hypothesis that DBS has excitatory modulation effect on neurons. The study provides guidance for accurately analyzing neuronal unit spike activity during high-frequency stimulation and also provides important clues for revealing the mechanisms underlying the treatment of brain diseases by DBS.]]> 2016/8/5 0:00:00 ZHU Yu-Fang, FENG Zhou-Yan, WANG Zhao-Xiang, YU Ying and GUO Zhe-Shan 396true 2016/8/5 0:00:00 LI Jun-Bei and QIU Xiao-Yan 395true 2016/8/5 0:00:00 YANG Long, XIONG Zhi-Yong, ZHANG Liang-Jun, FENG Xin-Chan, CHEN Kun, LI Yan, CHENG Ying, LONG Qing-Lin, XIAO Tian-Li, CHEN Lei, YAN Wen-Hui, LI Ling-Xin, CHAI Jin and CHEN Wen-Sheng 394true 2016/8/5 0:00:00 LIU Ji-Long, MIAO Zhi-Chao, LI Lei, XIAO Zhi-Xiong and CAO Yang 393true in vitro have been studied by UV spectra and atomic force microscopy (AFM) so as to reveal the structural basis of its anticancer activity. The structural characteristic after the incubation oxaliplatin at different concentrations with DNA for different time was investigated on freshly cleaved highly ordered pyrolytic graphite (HOPG) surface by atomic force microscopy (AFM). Based on the observed results, the interaction mechanism between oxaliplatin and DNA was proposed. High resolution AFM observation indicated that oxaliplatin can induce structural changes of pUC19 DNA from the extended chains to the intertangled structures with many nodes, and finally to a compact particle after 24 h incubation. The present results revealed that electrostatic and coordination interactions are important to condense plasmid DNA to a compact particle which may significantly influence the activity and cytotoxicity of oxaliplatin.]]> 2016/7/19 0:00:00 XU Feng-Jie, LIU Rui-Si, BIAO Lin-Hai, ZU Yuang-Gang and LIU Zhi-Guo 392true χ2-pos) was first proposed, which combined with pseudo position-specific scoring matrix (PsePSSM) and undirected composition of k-spaced amino acid pairs (Undirected-CKSAAP) were used to present protein sequences. Then 5 support vector machines models were constructed with the same proportion of positive and negative samples. At last, by weighted voting, our results showed that the prediction accuracy, Matthew’s correlation coefficient and area under ROC curve reached 89.62%, 0.79 and 0.96 respectively. They were superior to the literature report. It also demonstrated that the combination of three different features χ2-pos, PsePSSM and Undirected-CKSAAP has extensive application prospect in protein sites prediction such as glycosylation and phosphorylation.]]> 2016/7/18 0:00:00 XIANG Yan, CHEN Yuan, TAN Si-Qiao and YUAN Zhe-Ming 391true 1-42, TNF-α and INF-γ by ELISA. Results showed that bamboo leaf flavonoids significantly increased the proliferation of hippocampal neurons and suppressed hippocampal neurons apoptosis compared with isoflurane treatment only. The result of Morris water maze showed that bamboo leaf flavonoids significantly decreased average escape latent period and increased the frequency of crossing platform. The results of ELISA showed that bamboo leaf flavonoids suppressed the expression of Aβ1-42, TNF-α and INF-γ in hippocampus. Our findings suggested that bamboo leaf flavonoids enhanced immunity of aged rats and suppressed the expression of Aβ1-42, leading to the inhibition of isoflurane-induced neuronal apoptosis and cognitive impairment.]]> 2016/7/19 0:00:00 LIU Zhan and LIU Yan-Feng 390true Peg3 from sheep and analyzed the DNA methylation patterns in CpG islands and DMRs of the putative imprinted genes Peg3, Cdkn1c and Gtl2 in cloned lambs. We have provided evidence that Peg3 was highly methylated. The degree of methylation was 95.45% in the kidney and 88.18% in the lung of a natural sheep and 98.18% in the kidney and 87.27% in the lung for one cloned sheep. The bisulphite sequencing results for Cdkn1c show complete non-methylation (0%, 0.53%, 0.53%, 0.53%) in all samples. In addition, Gtl2 was hypomethylated in all lambs, from a linear correlation analysis, there were some differences in the quantitative values from both groups (correlation r2 = 0.77). These data show that the DNA methylation status of the three imprinted genes was similar in cloned and natural lambs.]]> 2016/7/18 0:00:00 WANG Feng, PAN Jing, ZHAO Li-Xia, LIUY Yi-Yi, ZHANG Li, WANG Shen-Yuan, LI Lu, ZHOU Huan-Min and ZHANG Dong 389true 2016/6/22 0:00:00 WEI Yan, WANG Yu-Jing, WU Bei-Bei, ZHANG Ying-Hao and HE Rong-Qiao 388true 2016/6/22 0:00:00 YUAN Hui-Ping, CHE Fu-Gang and YANG Ze 387true Tripterygium wilfordii Hook F, attracts increasing attention from researchers worldwide for its broad anti-tumor spectrum. Triptolide could bind to XPB (a subunit of the transcription factor TFⅡH) and leads to RNA polymerase Ⅱ-mediated transcription inhibition. Also, triptolide induces autophagy and apoptosis processes in cancer cells. Through regulating different kinds of related proteins, tumor repressor p53, plays key roles in the crosstalk between autophagy and apoptosis. Thus, in this study, the authors focused on triptolide induced p53-dependent autophagy and apoptosis processes in HeLa cells, and to demonstrate the regulation of crosstalk through p53-dependent transcription inhibitor and autophagy inhibitors treatment. Using CCK-8 assay and clonogenic assay, triptolide shows significant inhibition effect on HeLa cells. The apoptosis analysis by Western blot has shown that triptolide induces the cleavage of caspase3 and nuclear poly (ADP-ribose) polymerase (PARP). Immuno-fluorescence and Western blot assay have shown that triptolide also leads to LC3-Ⅱ accumulation and p62 degradations. Collectively, triptolide induces autophagy and apoptosis in a time- and dose-dependent manner. Meanwhile, p53 is significantly up-regulated and the mammalian target of rapamycin (mTOR) is down-regulated with triptolide treatment, and ultimately results in activation of autophagy and apoptosis. Western blot assay have shown that triptolide-induced autophagy and apoptosis are partly inhibited by utilizing p53-dependent transcription inhibitor (Pifithrin-α). Furthermore, the combination treatment of autophagy inhibitors with triptolide enhances triptolide induced apoptosis, and significantly improves inhibition effect of triptolide in HeLa cells. Triptolide induces p53-dependent autophagy and apoptosis in HeLa cells. p53 functions as a key regulator in the triptolide induced autophagy and apoptosis crosstalk.]]> 2016/6/21 0:00:00 ZHANG Bin, WANG Yun-Fei, ZHAO Fei, LEI Ming and HUANG Wei-Wei 386true ex vivo single myofibers isolated from weightless atrophied soleus. Finally, Smad3 knockout mice and its wild-type littermates were subjected to hindlimb suspension induced muscle atrophy, and the variability analysis demonstrated that the key role of Smad3 in regulating the plasticity of muscle satellite cells weightless muscle atrophy.]]> 2016/6/22 0:00:00 LI Jing-Long, LIU Hong-Ju, WANG Fei, HE Jian, LI Wen-Jiong, REN Wei and CHEN Xiao-Ping 385true Rad9, Rad1 and Hus1 are critical for the cell cycle checkpoint and can form a heterotrimer complex called 9-1-1 complex which was supposed to play important roles in the cell cycle checkpoint and other activities required for the maintenance of genome integrity. However, lack of high quality anti-Rad1 antibodies has seriously hindered the research on Rad1 as well as working mechanisms of the 9-1-1 complex at molecular level. In this study, a mouse anti-Rad1 monoclonal antibody (mAb) was successfully generated. The mAb possesses high affinity and specificity, and recognizes both endogenous mouse Rad1 (mRad1) and human Rad1 (hRad1) proteins and was successfully used in ELISA, Western blot analysis, immunoprecipitation and immunofluorescence assays. Using this mAb, we found that mRad1 protein expression was increased in Rad9+/+ mouse embryonic stem (MES) cells after hydroxyurea (HU, a genotoxic agent) treatment while not in Rad9-/- MES cells, suggesting that mRad1 expression is under Rad9 regulation. Furthermore, endogenous mRad1 was distributed mainly in the cytoplasm and did not migrate to the nucleus after HU treatment, contradicting the generally accepted hypothesis that Rad9, Rad1 and Hus1 form the 9-1-1 complex in the nucleus in response to genotoxic stresses. In summary, the exact molecular roles of Rad1 and the 9-1-1 complex are likely more complicated than previously expected and this anti-Rad1 mAb is a powerful tool for the future investigation on Rad1 as well as the 9-1-1 complex.]]> 2016/5/20 0:00:00 WANG Hai-Feng, AN Li-Li, SUN Shuang, CHEN Chuan, YE Chen, HANG Hai-Ying and ZHANG Xiu-Jun 384true 2016/5/20 0:00:00 ZHANG Xin-Min, ZHANG Rong, MENG Shu-Lin, XIE Wen-Jie and WANG Yi-Gang 383true 2016/5/20 0:00:00 SHI Lei, QIN En-Qiang, JIA Ya-Li, WU Dan, JIN Lei, ZENG Quan, JIANG Tian-Jun, NIE Wei-Min, YUE Wen and WANG Fu-Sheng 382true Xanthomonas campestris pv. campestris (Xcc) 8004. However, the protein encoded by XC_0119 shows 59% identity with fabV, the ENR from Pseudomonas aeruginosa, and contains the same catalytic triad Tyr-(Xaa)8-Lys. Expression of XccfabV restored the growth of the E. coli fabI temperature sensitive mutant JP1111 under non-permissive condition. In vitro assay identified that XccFabV catalysed enoyl-ACP with viarable chain lengths to acy-ACP, and the activity was not inhibited by triclosan. Furthermore, genetic research proved that XccfabV is an essential gene, and none of XccfabV deletion mutants was obtained. However, XccfabV in the chromosome could be deleted when plasmid expressing E. coli fabI was introduced into Xcc8004. And the fabI replaced mutant showed similar growth characteristic and fatty acid compositions with wild type, but changed to be sensitive to triclosan. These results domonstrated XccfabV is essential for growth, encodes ENR involved in fatty acid de novo biosynthesis, and FabV confers triclosan resistance on Xcc.]]> 2016/5/20 0:00:00 YU Yong-Hong, MA Jiang-Rong and WANG Hai-Hong 381true 2016/5/20 0:00:00 JIANG Zhi-Sen, YUAN Ding and PANG Lin 380true n=60). As a result, the three kinds of protein were mainly expressed in cell nuclei. Among them, NF-κB p65 and cyclinD1 were higher expressed in breast cancer tissues than in adjacent tissues while LXRα was lower expressed. Then, MTT assay was used to detect the proliferation of MCF-7 cells and Western blot was used to examine the expression of the three kinds of protein. TO901317 (a kind of artificial agonists against LXRs especially for LXRα) could increase LXRα expression, but decrease NF-κB p65 and cyclinD1 expression and suppress the proliferation of MCF-7 cells in a dose- and time-dependent manner (P < 0.05). Finally, the effects of LXRα siRNA and pyrrolidinedithiocarbamic acid (PDTC, an inhibitory of NF-κB) on TO901317 were observed respectively. LXRα siRNA could significantly decrease the up-regulation of LXRα expression and reverse the inhibited effect of TO901317 on cyclinD1 and NF-κB p65 expression and MCF-7 cell proliferation (P < 0.05) while PDTC could strengthen the inhibition of cell proliferation and further down-regulate NF-κB p65 and cyclinD1 expression induced by TO901317 (P < 0.05), but have little effect on LXRα. In a conclusion, the expression of LXRα, NF-κB p65 and cyclinD1 plays an important role in the proliferation of human breast cancer cells, so as to provide a new method for the molecular targeting treatment of breast cancer in the future.]]> 2016/3/21 0:00:00 TU Jian, LIU Xiao-Wang, LI Tao, YU Ping, DING Wei-Ke, LU Kai-Qiang, CHEN Xiao-Xiao, PENG Lu and ZHOU Zhi-Gang 379true 2016/3/21 0:00:00 ZHANG Jin, SHE Qiang, FAN Jie and WEI Fei-Yu 378true in vivo because the invasive measurement would disturb the cellular local structure and physiological activities, and the non-invasive measurement couldn't provide information about myosin Ⅱ activity accurately. In this paper, we developed and applied new images analysis methods, such as protein fluorescent image tracking and image correlation spectroscopy, to quantify the kinetics of disassembly and reassembly of actomyosin networks and compared them to studies by other groups. This analysis suggested the following processes contribute to the assembly of cortical actomyosin and stress fibers: random myosin mini-filament assembly and disassembly along the cortex; myosin mini-filament aligning and contraction; stabilization of cortical myosin upon increasing contractile tension. We found that the number of myosin Ⅱ and focal adhesions are very important to the formation and stability of the typeⅠ, Ⅱ and Ⅲ actomyosin network in HeLa cells, and the activity of myosin Ⅱ, which determines the dynamics of the actomyosin network reorganization, can be quantified through STICS (spatial temporal image correlation spectroscopy). The formation of typeⅠ, Ⅱ, and Ⅲ actomyosin networks was explained through a mechanical model by adjusting the parameters of myosin Ⅱ activities and number density. The STICS method used in this study can be applied to evaluate the activity of other proteins in live cells.]]> 2016/3/21 0:00:00 SHI Xiao-Chuan, SHEN Chao, NIE Wei, LIU Feng and MENG Xiao-Liang 377true 2016/3/21 0:00:00 CHU Wen-Dan, ZHOU Cui-Yan, LU Ya-Fei, XU Yang and LI Wen-Qi 376true Caenorhabditis elegans, the expression of all piRNAs, small of miRNAs were influenced by the mutation of prg-1. Especially, some ncRNA-like, a kinds of small RNA, were found in C. elegans. The ncRNA-like had the similar sequence with piRNAs or miRNAs. The results of the analysis in the gene sites between the ncRNA-like and piRNAs or between ncRNA-like and miRNAs, showed that ncRNA-like had the same gene sites with piRNAs or miRNAs. All of these showed that the ncRNA-like were a kind of small RNA and may be processed from the RNA precursor same to piRNAs or miRNAs.]]> 2016/3/21 0:00:00 HUANG Da-Wei, CUI Dong-Ya, WANG Jia-Jia, HE Shun-Min and ZHANG Peng 375true 2016/3/21 0:00:00 JIANG Min, LI Heng, WANG Miao, SHI Jin-Song, XU Zheng-Hong, LUO Chun-Qin and MA Yan-He 374true in vitro by okadaic acid (OA), and demonstrate that IL-6 maybe the key protein to effect the recovery function of hUC-MSCs to protect the injured cells.]]> 2016/2/19 0:00:00 ZHAI Jing-Lei, CAO Ning, YUE Wen, JIA Ya-Li and PEI Xue-Tao 373true P < 0.05) in myocardial cells after hypoxia/reoxygenation injury. Moreover, myocardial apoptosis was aggravated according to flow cytometry. According to RT-PCR and Western blot, p62 was down-regulated whereas beclin-1 was up-regulated in cardiomyocytes (P < 0.05) after hypoxia/reoxygenation injury. These consequences inditicated that hypoxia reoxygenation induced abnormal myocardial apoptosis and autophagy. miR-21 mimic or inhibitor were transfected into cardiomyocytes via liposome. MiR-21 mimic transfection significantly enhanced myocardial apoptosis (P < 0.05), up-regulated expression of p62, and down-regulated of expression of beclin-1in myocardial cells in rat cardiomyocytes(P < 0.05), while miR-21 inhibitor transfection caused opposite effects. These data suggested that miR-21 in rat cardiomyocytes exposed to myocardial hypoxia reoxygenation injury can accelerate cell apoptosis and inhibit cell autophagy. Bioinformatics projections shown that Rab11a 3'-UTR contains a binding site for miR-21. Dual luciferase report gene assay system, coupled with the overexpression of Rab11a assay validated that Rab11a is one of miR-21 target genes. Overexpression of Rab11a significantly attenuated myocardial apoptosis，up-regulation of p62 and down-regulation of beclin-1 induced by miR-21in hypoxia/reoxygenation injury. In conclusion, miR-21 can promote myocardial apoptosis and inhibit myocardial autophagy by negative regulating Rab11a in rat cardiomyocytes exposed to hypoxia/reoxygenation injury.This research proposes a new strategy for the prevention and treatment of myocardial ischemia-reperfusion injury.]]> 2016/2/19 0:00:00 LI Hui, ZHAO Kai and WANG Lei 372true 2016/2/19 0:00:00 QIAN Chen-Can and LIU Zu-Xiang 371true Caenorhabditis elegans. Determining the optimal focus in an optical microscope is based on a clarity-evaluation function that is applied to images acquired from different focuses of the same field. The maximum value of the function is considered as the point of optimal focus. In this paper, 16 autofocus algorithms which were collected from well-known algorithms as well as the most recently proposed focusing algorithms have been evaluated. Through these evaluations, an optimal algorithm was found for C. elegants lipid droplets to set up an automatic screening system. Many features were assessed in this paper, for instance accuracy, computational time, addition of noise, and focusing curve. Our results have shown that most of the algorithms show an overall high performance for this type of image, and absolute Tenengrad algorithm will be our first choice for its best performance considering accuracy.]]> 2016/2/19 0:00:00 ZHANG Xiang, JIA Ce and XIE Kang 370true 2016/1/19 0:00:00 ZHAI Yan-Hong, LIU Chang-Mei, GUO-Hui, YANG Gang-Long and GUAN Feng 369true 2016/1/19 0:00:00 ZHANG Fei, WANG Shi-Xiang, WANG Ling and SONG Kai 368true 2016/1/19 0:00:00 CAO Zan-Xia, DONG Chuan, ZHAO Li-Ling and WANG Ji-Huan 367true Ceratosolensolmsi, generated five expression modules, which were flagged as blue, turquoise, brown, green and yellow. These involved two female-biased expression modules and three pupa-biased expression modules, respectively. Gene ontologies indicated three functional enrichment gene sets in modules turquoise and yellow. Two functional enrichment gene sets that participate in cell cycle or have nucleotide binding activityclustered in turquoise module. The functionally enriched gene set in yellow module played roles in cell differentiation, especially in neuron morphogenesis.]]> 2016/1/19 0:00:00 CUI Dong-Ya, SUN Xu-Bin, WANG Jia-Jia, ZHANG Peng, SUN Bao-Fa, CHEN Xiao-Wei, Robert W. Murphy, HE Shun-Min and HUANG Da-Wei 366true t test results show that 6 lectins (STL, WGA, BPL, DBA, PTL-Ⅰ and MAL-Ⅰ) showed different binding signals between two groups (P < 0. 05). Among these, STL, WGA, BPL and DBA exhibited increased binding signals in the MCN cyst (the ratio value > 2.0), which indicated the abundance of Tn antigen in MCN group was higher than that in SCN group. It may correlated with a significant increase in the mucin secretion of cancer epithelial cells. Conversely, the glycopatterns of GalNAcα-1, 3Gal and Galβ-1, 4GlcNAc identified by PTL -Ⅰ and MAL -Ⅰ were decreased in MCN group when compared with SCN group (ratio values < 0.5). In order to investigate precisely alterations of glycopatterns associated with MCN and SCN, SDS-PAGE and lectin blotting analysis was performed with STL and BPL staining. The binding signals of this glycoprotein were significantly higher in MCN than that in SCN groups.]]> 2016/12/22 0:00:00 WANG Ying, SUN Yu-Fa, CHAI Ning-Li, XU Wei, FENG Jia, WANG Xiang-Dong, TANG Ping, YANG Jing, ZHANG Wei and LINGHU En-Qiang 365true 3+) with larch tannin according the liquid phase synthesis method, and these complexes were characterized with FT-IR, XPS, UV and coordination number determination method. The antifungal activity of Re3+-LT against Aspergillus niger, Monascus anka, Phanerochaete chrysosporium and Actinomucor elegans was evaluated in vitro by using the oxford cup and agar dilution assay. It was found that the Re3+-LT complexes are effective for these four fungi, and the antifungal activity was in the order of Ce3+-LT > Gd3+-LT > La3+-LT > Nd3+-LT > Yb3+-LT. The minimum inhibitory concentration of Ce3+-LT to these four fungi is 1.6 g·L-1, 1.6 g·L-1, 0.8 g·L-1, and 1.6 g·L-1, respectively, while the Yb3+-LT is 3.2 g·L-1, 1.6 g·L-1 3.2 g·L-1, and 3.2 g·L-1, respectively. In terms of fugicide, Yb3+-LT is the best one and the minimum fugicide concentration of Yb3+-LT to these four fungi is 6.4 g·L-1, 3.2 g·L-1, 3.2 g·L-1, and 6.4 g·L-1, respectively. In addition, the fugicide activity of Nd3+-LT and Gd3+-LT against Aspergillus niger and Actinomucor elegans is less powerful even they have good antifungal activity.]]> 2016/12/22 0:00:00 TIAN Cui-Cui, ZENG Wei-Cai, LI Xia, LIAO Xue-Pin and SHI Bi 364true etc. FOXO1 is a vital member of the Forkhead family of transcription factors known to regulate the transcription of genes involved in cell cycle arrest, DNA repair in response to oxidative stress or apoptosis. However, it is unclear how the NO effect on FOXO1. In this study, we observed vital role of FOXO1 dependent transcriptional activation on neuronal death in response to Nitric oxide over-production. We found that NO donor GSNO or L-Arginine significantly increased the FOXO1 transcriptional activity, which induce the FOXO1 downstream proapoptotic genes (FasL, Bim) expression and finally induce neuronal death. In addition, we screen natural Chinese herb extracts targets to regulating FOXO1 activity. Emodin shows dramatically effect in suppression of FOXO1 transcription activity and protein levels. What’s more, Emodin can attenuate neuronal death induced by L-Arg. The current study first demonstrate that Nitric Oxide could regulate FOXO1 transcriptional activity to damage neuronal cell, which will benefit to deep understanding the neurotoxicity of NO free radical. Secondly, by Chinese herb extracts and antioxidants screen, we identified Emodin as one FOXO1 activity modulator which can attenuate neurotoxicity induced by NO. The current study will also provide a new insight for explaining the mechanisms of pathology of neurodegenerative disease and neuroprotective effects of Emodin.]]> 2016/11/21 0:00:00 CHEN Su-Ling, ZHOU Jie-Chao, ZHANG Jie, ZHUANG Jiang-Xing and LIU Ya 363true ChCel5A and substrate was quantitatively analyzed using fluorescence spectrum. The substrate concentration range when tryptophan can specifically bind the substrate was determined and the change of binding constant caused by a single amino acid mutation in active-site architecture of ChCel5A was measured through quantitative analysis of tryptophan fluorescence quenching. Compared with the measurement of kinetic parameters, fluorescence spectroscopy could accurately characterize the binding affinity between cellulase and substrate, as well as the change of kinetic parameter caused by a single residue mutation. Besides, when using pNPC as the ligand, the binding constant could be overestimated for 20-100 times due to the increase of strong electrophilic groups. Fluorescent spectrometry can measure the kinetic parameter of cellulase-substrate binding process in a sensitive and rapid way, providing a new perspective for the quantitatively analyzing the interaction between proteins and substrates.]]> 2016/11/22 0:00:00 WANG Pei-Pei, ZHANG Li-Li, ZHANG Qing, WANG Lu-Shan and CHEN Guan-Jun 362true 2016/10/24 0:00:00 ZHANG Xiang, LI Min and JIA Ce 361true fabH genes (Bacillus subtilis BsfabH1 and BsfabH2, Staphylococcus aureus SafabH and Streptomyces coelicolor ScofabH), and three homologous Gram-negative bacterial fabH genes (Escherichia coli EcfabH, Ralstonia solanacearum RsfabH, and Xanthomonas oryzae pv. oryzae XoofabH) for further study. Expression of each of the seven fabH genes restored the growth of the R. solanacearum fabH mutant RsmH in the absence of octanoic acid, indicating that all encoded FabHs have 3-ketoacyl ACP synthase Ⅲ activity. Furthermore, fatty acid profiling showed that complementation with the four Gram-positive bacterial fabH genes or the Gram-negative XoofabH rendered the R. solanacearum fabH mutant able to produce a large quantity of BCFAs, while the RsmH mutant harbouring EcfabH or RsfabH could only synthesize straight chain fatty acids, suggesting XooFabH performs a role different from EcFabH or RsFabH in BCFA sysnthesis. In vitro analysis showed that, similar to the four Gram-postive FabHs, XooFabH has a preferance for using branched chain acyl-CoAs as primers. XooFabH also displayed a strong activity towards short or medium chain acyl-CoAs (~C4-C10), unlike other Gram-positive FabHs. These results confirmed that FabH from different bacteria have different biological characteristics, and display a preferance for branched chain acyl-CoAs, which are essential for BCFAs synthesis.]]> 2016/10/24 0:00:00 YU Yong-Hong, MA Jian-Rong, MIAO Xin-Yu and WANG Hai-Hong 360true via a “kindling” effect. It can also produce changes of synaptic plasticity that may persist. However, a long HFS train with a duration several minutes or longer can be safely applied in deep brain stimulation to treat various brain diseases in clinic. Therefore, we speculate that a sustained HFS could change the neuronal responses induced by a short HFS. To verify this hypothesis, 100 Hz HFS with durations of 5 s and 2 min were applied to the Schaffer collateral of afferent fibers in the hippocampus CA1 region of rats. The response potentials of downstream populations of neurons, i.e., population spikes (PS), evoked by a single test pulse were monitored after the termination of HFS trains. The evoked-PS potentials following the two types of stimulation were compared. The results showed that after-discharge events with epileptiform activity appeared immediately following 5 s short HFS trains. In addition, the changes of amplitude and latency of the evoked-PS suggest an increase of excitability persisting for tens of minutes after the termination of 5 s short HFS. In contrast, silent periods of a few tens of seconds without any neuronal firing appeared immediately following 2 min long HFS trains. Furthermore, the amplitude of evoked-PS by test pulses recovered to the baseline level in a few minutes after the termination of long HFS. Because long HFS trains include short HFS trains, these results indicate that the late stimulation of long trains can change the effects on the downstream neural networks produced by their early phases and eliminate the long-term excitatory effects induced by short trains. These findings are of significance for further revealing HFS mechanisms and for advancing the clinical applications of deep brain stimulation.]]> 2017/9/21 0:00:00 ZHOU Wen-Jie, FENG Zhou-Yan, QIU Chen and MA Wei-Jian 359true pro was established and a small-molecular compound library was screened. The virus EV71-MZ was isolated from an HFMD patient, and identified by PCR. A 3Cpro recognition site was inserted into the middle region of YFP open reading frame to generate the mYFP by insertion mutation. The full length mYFP proteins were observed by fluorescence microscope and the protein level was measured by using microplate reader in Ex(500 nm)/Em(535 nm). Change of the fluorescence value reflected the degree of the inhibition on 3Cpro activity. A small-molecular compound was screened by using the established screening model in the high throughput drug screening system, then the antiviral activity of the active compounds was further evaluated by plaque assay. As a result, mYFP expressed well in 293A cell; the expression of 3Cpro reduced the fluorescence signal remarkably, however, the signal was recovered by adding Rupintrivir, an inhibitor of 3Cpro. These results indicated that the screening model targeting 3Cpro was established successfully. 26 of 26 000 compounds significantly reverted the fluorescence signal of the mYFP in the presence of 3Cpro; noticeably, two of the 26 compounds, i.e. numbering 3 and 8, exhibited strong antiviral activity by plaque assay. All together, 3C-mYFP co-expression system is an optimized and effective screening method for high-throughput screening of anti-EV71 3Cpro drugs.]]> 2017/9/21 0:00:00 ZENG Shi-Nuan, LI Qian-Wen, PAN Ting, MENG Xiao-Bin, HUANG Qing-Yuan and GUO Xue-Min 358true 2017/9/21 0:00:00 LI Gao-Feng, HU Guo-Dong, ZHANG Chen, JI Bao-Hua and WANG Ji-Hua 357true via imperfect complementary target base pairing．We confirmed miR-21 regulates PTEN and TPM1 protein expression without significantly affecting PTEN and TPM1 mRNA expression via imperfect complementary target binding in HEK293T and HeLa cells. Furthermore, Overexpressing miR-21 could significantly shorten half-lives of PTEN and TPM1, miR-21 enhanced PTEN and TPM1 decay．Cells were transfected with lncRNA-GAS5 expression vector, we found lncRNA-GAS5 competitively bound miR-21 and increased the half-lives of PTEN and TPM1. Besides, miR-21 bound with lncRNA-GAS5 could lead rapid decay of lncRNA-GAS5. This study indicates lncRNA-GAS5 functions as a miR-21 “sponge” that inhibits mRNA degradation of the miR-21 imperfect complementary targets PTEN and TPM1, and miR-21 also could regulate lncRNA-GAS5 stability by base pairing．Further exploration of the mechanisms may improve our understanding of the lncRNA-miRNA-mRNA sophisticated regulatory feedback loop.]]> 2017/7/17 0:00:00 ZHENG Wei, DONG Jie, LI Shao-Hua, DING Hong-Mei, LI Hui, HUANG Ai-Xue, XIA Wei, BAI Chen-Jun, GUO Xiao-Hua, LI Da, GENG Jie, LI Jie and SHAO Ning-Sheng 356true 2017/7/17 0:00:00 WU Zhang-Song, LUO Zhi-Qiang, ZHANG Ming-Yu, LIAN Cui-Hong, LIN Xin-Yan, LIANG Yu-Hong, GU Hong-Sheng and ZHU Yan-Xia 355true 2017/7/19 0:00:00 WANG Jing, ZHOU Jun, MO Wei-Chuan, HE Ying-Ge, WEI Yan, HE Rong-Qiao and YI Fa-Ping 354true 2017/7/18 0:00:00 ZHANG Di, Vincent Nourrit, SANG Xin-Zhu and Jean-Louis de Bougrenet de la Tocnaye 353true Cre-miR914 were down-regulated significantly under multiple stresses (heat shock, UV-B and salinity) in Chlamydomonas reinhardtii through Q-PCR screening experiments, and bioinformatics analysis showed that the target gene of Cre-miR914 may be RPL18. But the functions of Cre-miR914 and its target gene in heat shock adaptation are unclear, this study addressed these issues through multiple experiments. In this study, we identified the target of Cre-miR914 through bioinformatics and degradome sequencing, and validated expression of Cre-miR914 and RPL18 under heat shock through Q-PCR. Then we constructed cell lines of Cre-miR914 overexpression and RPL18 overexpression for further study. And finally we performed stress adaptation experiments under heat shock stress to check the function of microRNA and its target in stress adaptation, which includes cell growth assay, cell vitality counting, reactive oxygen species (ROS) production and lipid peroxidation (MDA) measurements. Bioinformatics and degradome sequencing indicated the target of Cre-miR914 is RPL18/i>; Q-PCR results showed that Cre-miR914 expression reduced under heat shock, but RPL18 expression increased, which confirmed our previous results of screening experiment. Then we got more than 3 cell lines with overexpressing of Cre-miR914 and RPL18. Further growth experiment under heat shock indicated that Cre-miR914 overexpression lines had a lower growth than the wild-type line (cw15), while RPL18 overexpression lines had a higher growth than the wild-type line (cw15). Cells vitality (photosynthesis activity) experiment under stress also demonstrated that Cre-miR914 overexpression lines had a lower vitality than the wild-type line (cw15), while RPL18 overexpression lines had a higher vitality. The cell damage (ROS production and MDA content) experiments showed that there were more cell damages (ROS production and MDA content) in Cre-miR914 overexpression lines than the wild-type line (cw15), while that of RPL18 overexpression lines were lower than the wild-type line (cw15). These results illustrated that overexpression of Cre-miR914 reduced heat shock resistance ability in algae, while overexpression of RPL18 increased heat shock resistance ability. We maybe discovered a new regulation mechanism of heat shock adaptation in algae, in which Cre-miR914 and its target gene RPL18 are engaged in adaptation regulation to heat shock in Chlamydomonas reinhardtii.]]> 2017/6/20 0:00:00 ZHANG Feng-Ge, WANG Bo, WANG Xiao-Lin, LI Xiao-Yan, LI Gen-Bao, LI Dun-Hai, SHAN Ge and WANG Gao-Hong 352true in vitro and inhibited the growth of prostate xenograft tumor in vivo. We have processed a gene expression microarray assay to investigate the concrete mechanism of miR-26b inhibition on prostate cancer cells proliferation and migration. We found that miR-26b significantly up-regulated 57 genes expression level, and simultaneously down-regulated 55 genes expression (fold change >2; P < 0.05) in PC-3. The differential genes were most associated with the regulation process of cell proliferation, apoptotic process, protein phosphorylation and ubiquitination respectively, and enriched in multiple pathways including TNF signaling pathway and TGF-β signaling pathway. Among these filtered genes, CEACAM6 was significantly down regulated by miR-26b with a 2.17-fold. We identified a putative miR-26b binding site on 3′UTR region of CEACAM6 and validated that miR-26b bound to the 3′UTR region of CEACAM6 mRNA, suggesting that CEACAM6 is a direct target of miR-26b. Our results suggest that miR-26b suppresses cell proliferation by targeting CEACAM6 in PCa cells and miR-26b may be a candidate tumor-suppressor in prostate cancer.]]> 2017/6/20 0:00:00 Daguia Zambe John Clotaire, WANG Yun-Fei, HUANG Wei-Wei, ZHAO Fei, ZHANG Bin and LEI Ming 351true Mycobacterium tuberculosis is a successful pathogen of human being that causes about millions people death annually. M. tuberculosis posses an unique cellular envelop which can protect itself from the attack of immune system and harsh environment. The mycobacterial cell wall consists of inner membrane, peptidoglycan/arabinogalactan layers, a mycolic acid-containing layer, and the outer capsular layer. ESX secretion system is involved in transporting substrates through cellular envelop to extracellular environment. ESX secretion system has five pathways: ESX1-5. ESX-1 pathway contributes most in pathogenicity of M. tuberculosis and became a continuing concern on interaction between M. tuberculosis and host. EspB shares a co-dependent secretion way with ESAT-6, a key virulence of ESX-1which is responsible for the escape of M. tuberculosis from host cell to cytoplasm. We solved the crystal structure of EspB from M. tuberculosis. EspB posses conserved WxG domain and YxxxD domain, and so does other substrates of ESX-1 pathway. Research on EspB shows that it can interact with a component of secretion apparatus-EccC. We suppose that WxG domain and YxxxD domain may function together as a signal peptide for the recognition of secretion apparatus during the secretion of EspB. In addition, EspB heptamer has a relative rigid structure and a large enough inner-diameter for the transport of ESAT-6/CFP-10 dimer, which makes it possible to function as a channel for ESAT-6/CFP-10.]]> 2017/5/22 0:00:00 TANG Xiao-Meng and WANG Da-Cheng 350true in vitro and underlying mechanism remain to be investigated. Here, we compared IL-2 expression levels in peritumoral tissue from HCC patients between HBV-positive group and HBV-negative group. We then overexpressed HBV x protein (HBx) in immortalized human liver cell line THLE-2, and mimic microenvironmental oxidative stress with hydrogen peroxide (H2O2) treatment in vitro. We demonstrated that hepatocytes can express IL-2 in vitro through MAP3K7/NF-κB pathway in the presence of both HBx stimulation and mild oxidative stress. Targeting microenvironmental oxidative stress offers a promise for prevention and therapy of HBV-associated HCC metastatic recurrence.]]> 2017/5/22 0:00:00 GAO Dong-Mei, ZHAO Yue, CHEN Jie, ZHAO Yan, CHEN Jun and ZHOU Hai-Jun 349true χ2-statistics of the residuals in least-square fits of the models to the image data. By comparing the detection accuracy with or without the “self-checking” algorithm in simulated images, we found that the detection accuracy with the “self-checking” algorithm was improved significantly. Meanwhile, we proposed an optimized flow chart of vesicle tracking which was applied to analyze the vesicles in mice β cells. We found that the number of vesicle traces will increase and the average docking time of vesicles will decrease after glucose stimulation based on our tracking analysis. This is because β cells will release insulin to regulate glucose balance with the help of vesicle translocation and secretion after glucose stimulation. In a word, we quantified the vesicles activity in mice β cell by tracking analysis on subcellular level.]]> 2017/5/22 0:00:00 ZHANG Xiang, LIU Xin-Yi, LV Ping-Ping and JIA Ce 348true P<0.001). The recombinant virus could induce apoptosis of hepatocellular stem-like cells through caspase pathway. Therefore, the recombinant adenovirus Ad.wnt-E1A (Δ24 bp)- TSLC1 may be a promising therapeutic agent for targeting hepatoma stem cells, which has certain application value to clinical treatment.]]> 2017/4/24 0:00:00 ZHANG Jian, LAI Wei-Jie, LI Qiang, JIN Jin, XIAO Bo-Duan, GUO Wan and WANG Yi-Gang 347true via glycosylation needs further illustrated. In this study, the expression of glycosyltransferases were analyzed using glycan-related gene chip in miR-10b-overexpressed human normal mammary epithelial cells MCF10A. Then, the alteration of N-glycan and O-glycan was analyzed using the methods established in our laboratory. The expression of glycosyltransferase gene Fut8, MGAT3 and OGT and corresponding glycans were evaluated by qRT-PCR, Western blotting and lectin blotting. Taken together, our results provide the theoretical basis of glycomics on the role of miR-10b in breast cancer.]]> 2017/4/24 0:00:00 GUO Dong, GUO Jia, WU Yan-Li and GUAN Feng 346true S = 68.8°/RIU, and the surface sensitivity is S = 1.5°/(mol·L-1). The Raman enhancement factor of the dual-mode biochip reaches up to 106 and the minimum detection concentration of 10-15 mol/L could be achieved. Due to its high-fidelity measurement of the dual-mode chip and the high precision of microscopy imaging technology, multi-channel label-free detection is expected to be widely used in biological detections. Our future work is to achieve the practical application of SPR-SERS dual-mode biological detection.]]> 2017/4/24 0:00:00 liwenxue, shiyuanzhen, jiangzhisen, suyarong and panglin 345true PPARγ, C/EBPα, FABP4 and FATP1). In addition, silencing the PI3K gene by siRNA transfection also inhibited adipogenic trans-differentiation in C2C12 cells. Moreover, suppressing both activation and expression of PI3K/Akt induced apoptosis. Taken together, our findings indicated that the PI3K/Akt pathway plays a key role in adipogenic trans-differentiation of myoblasts.]]> 2017/3/22 0:00:00 QI Ren-Li, HUANG Xiao-Feng, WU Yong-Jiang, WANG Jing, LIU Hong, HUANG Jin-Xiu and WANG Qi 344true 121-138 were systematically investigated using fluorescence anisotropy techniques. As a result, the equilibrium dissociation constant of Mad2 two folds binding with Cdc20121-138 were both within 10-6 mol/L range in lower ionic strength solutions, and the KD value of C-Mad2 and Cdc20121-138 was 5 times lower than that of O-Mad2. While in high ionic strength solution, there was no obvious difference on KD value of C-Mad2 and O-Mad2 binding with Cdc20121-138. The kinetic experiments suggested that the dissociation rate constant (kd) between C-Mad2 and TAMRA-Cdc20121-138 was similar to that of O-Mad2, but the association rate constant (ka) between C-Mad2 and TAMRA-Cdc20121-138 was one order of magnitude higher than that of O-Mad2, which suggested that the binding of C-Mad2 with Cdc20121-138 is thermodynamically more stable and kinetically faster. Studies on interactions between Cdc20 mutants and Mad2 together with influence of ionic strength on their interactions both suggested that the interaction of Mad2 and Cdc20 is possibly not achieved by electrostatic interaction, but through hydrophobic interactions. Our results provide key information for revealing the conformational transition mechanism of metamorphic proteins and their important role in mitosis.]]> 2017/3/22 0:00:00 ZHANG Hui-Ting, ZHAO Yuan-Yuan, GE Bao-Sheng and HUANG Fang 343true 2017/3/22 0:00:00 ZHOU Dan-Dan, YU Yan-Qing, WU Hao, LI Yan-Ni and QIAO Jian-Jun 342true Shigella flexneri was the most common pathogen causing bacillary dysentery. Thioredoxin-dependent thiol peroxidase (SF2523) proteins was from Shigella flexneri 2a str. 301. It belonged to thioredoxin peroxidase family and played an important role in protecting the biological macromolecule by scavenging active oxygen generated in the process of aerobic metabolism. To understand the underlying mechanism, prokaryotically expressed thioredoxin-dependent thiol peroxidase protein was purified using affinity chromatography and gel filtration, crystallized using the vapour-diffusion method. The crystal grew in a condition consisting of 1.8 mol/L tri-Ammonium citrate, pH 7.0 using 1 g/L protein solution at 289 K. A complete data set was collected from a crystal to 2.75 ? resolution using synchrotron radiation at 100 K. The crystal belonged to space group P212121, with unit-cell parameters a = 35.80 ?, b = 50.63 ?, c = 88.52 ?, α = β = γ = 90.00°. One molecule was found in the asymmetric unit with a Matthews coefficient of 2.03 ?3/u, corresponding to a solvent content of 39.56%.]]> 2017/3/22 0:00:00 LIU Yong, LU Fang, GUO Gang-Xing, FENG Duo, ZHANG Bei, GAO Wei and BI Ru-Chang 341true Cucumber mosaic virus (CMV) as a vector expressing a TM sequence effectively inhibited the activity or stability of somemiRNAs in plants, thereby impaired their regulation to the targets．However, the miRNAs binding to their TM sequences carried by CMV inhibited accumulation of the virus to some extent. To analyze underlying reason of the inhibition to virus accumulation when a miRNA binds to its TM sequence in CMV genome, we analyzed influence of various miRNA TM sequences in CMV genome on virus accumulation using RNA blotting．Then, using GFP as a reporter gene, effect of the TM sequences on virus accumulation was analyzed by fluorescence microscopy, Western blotting and RNA blotting．Subsequently, using GFP as a reporter gene again, we analyzed effect of the TM sequences on GFP translation．Finally, the influence of the TM sequences on virus negative-strand RNA synthesis was tested using CMV trans-replication system．The results showed that targeting of five plant endogenous miRNA species tested to their TM sequences carried by CMV genome inhibited virus accumulation to a varied extent．Binding of the miRNAs to their TM sequences suppressed translation of the GFP protein and synthesis of the negative strand. Plant endogenous miRNAs targeting to their target mimics carried by viral genome inhibits translation of viral protein and synthesis of negative strand, thus reduces virus accumulation．The data present here provide possibility for developing a new antiviral methods.]]> 2017/2/22 0:00:00 WANG Mei-Mei, CHEN Wen-Hu, LIAO Qian-Sheng and DU Zhi-You 340true 2+ released from the inositol 1, 4, 5-trisphosphate (IP3) receptor channel in the regulation of the phosphorylation-dephosphorylation cycle process involved in glycogen degradation by glycogen phosphorylase have theoretically investigated using stochastic the Li-Rinzel model for cytosolic Ca2+ oscillations. The results show that the fraction of active phosphorylase is decreased with the total number of receptor channel IP3R increasing and for the small concentration of IP3 with appearance of coherence resonance.]]> 2017/2/22 0:00:00 Anvar ROZI, Ablet MAMAT and Askar ANVAR 339true 2017/1/17 0:00:00 WU Yan-Li, YANG Gang-Long, MIAO Ming-Yong and GUAN Feng 338true -8～10-6 s, and the voltage of intracellular membranes will exceed the epicyte’s. The result also indicates as t increases, the outer and inner membrane field intensity decreases but bounce back when t is about 3.2×10-7ns of parameters group, which means this is a minimum and is a very meaningful value for the following research. The Window Effect provides a research approach to accounts for biomedicine effect about electrical pulses.]]> 2017/1/17 0:00:00 WANG Yue, ZHOU Xiao-Yang, LI Ye-Feng, SUN Jiu-Xun and YANG Hong-Chun 337true Mycobacterium tuberculosis is the pathogen of tuberculosis which causes about millions people death annually. M. smegmatis, as a type of non-pathogenic strain of Mycobacterium closely related to M. tuberculosis, is the most studied model strain in laboratory. Mycobacterium encodes three types of chromatin proteins, i.e. histone-like protein HU, Lsr2 and integration host factor (IHF). To investigate the functional roles of IHF in chromosomal DNA organization, we expressed and purified IHF protein from M. smegmatis (MsIHF) in Escherichia coli, and analyzed the effects of MsIHF on DNA topology in vitro. MsIHF exists as a stable homodimer in solution. MsIHF exhibits preferred binding to negatively supercoiled DNA rather than linear or relaxed DNA. This protein is also capable of constraining negative DNA supercoils. Further analyses showed modulations of topoisomerase activities by MsIHF in vitro. MsIHF obviously inhibits the relaxation of supercoiled DNA by topoisomeraseⅠ from E. coli. By contrast, this protein slightly stimulates E. coli gyrase to introduce negative supercoils into relaxed DNA. These data suggests that MsIHF may alter the topological structure of chromosomal DNA through modulation of the activities of topoisomerases, and therefore regulates the organization of chromosome.]]> 2017/12/19 0:00:00 CHEN Yuan-Yuan, ZHANG Xian-En and BI Li-Jun 336true χ2-pos, and then integrates this feature with the pseudo position-specific scoring matrix (PsePSSM). A Support Vector Machine (SVM) classifier with balanced positive and negative samples was created, and the S, T, Y independent testing results for the Matthew correlation coefficient, the inferior surface integral of the ROC curve and the precision were (0.59, 0.87, 79.74%), (0.55, 0.85, 77.68%) and (0.50, 0.81, 75.22%), respectively, which are significantly superior to the results reported previously. The integration of the χ2-pos and the PsePSSM offers a promising method to predict phosphorylation sites more accurately in proteins.]]> 2017/12/19 0:00:00 TAN Si-Qiao, LI Qian, CHEN Yuan and PENG Jian 335true Bacillus megaterium, and its mutant (A74G, F87V, L188Q, D168H) could hydroxylate indole to indigo in Escherichia coli whereas the bioconversion yield is low under normal culture conditions (37℃, 250 r/min). Chaperones from hyperthermophilic archaea Pyrococcus furiosus were co-expressed with P450 BM3 mutant in E. coli to investigate whether they could improve the production of indigo, and results indicated that prefoldin could significantly increase the indigo production. Our research also revealed that enzyme activity was not the restrictive factor in the indigo bioconversion process. We also demonstrated that prefoldin could increase the intracellular nicotinamide adenine dinucleotide phosphate (NADPH)/NADP+ ratio, and this ratio was probably related to the increase of the indigo bioconversion yield.]]> 2017/12/19 0:00:00 PENG Shuai-Ying, CHU Zhong-Mei, LU Jian-Feng, LI Dong-Xiao, WANG Yong-Hong, YANG Sheng-Li and ZHANG Yi 334true in vitro. The transcriptional factor inhibitor of DNA binding 1 (Id1) can promote mESC self-renewal in serum condition without LIF. However, whether the other Id family members, such as Id2 and Id3, have similar function in mESCs is still unclear. In this study, we overexpressed Id2 and Id3 in 46C mESCs respectively, and found that both factors are able to support mESC self-renewal in the absence of LIF, while the self-renewal-promoting effect of Id2 is stronger than Id3. RNA-sequence approach revealed that Id2 is capable of upregulating c-Myc and n-Myc expression levels. Functional studies demonstrated that knockdown of c-Myc and n-Myc together could greatly reduce the function of Id2 in sustaining the undifferentiated state of mESCs, indicating that Myc family members have functional redundancy and can mediate the self-renewal-promoting effect of Id2 in mESCs. Our results will expand the current understanding of the pluripotency regulation network of embryonic stem cells.]]> 2017/11/20 0:00:00 GUO Meng-Meng, YING Qi-Long and YE Shou-Dong 333true 3. The mRNA expression of C3aR was analyzed by quantitative real-time PCR; the protein level of C3aR was evaluated by Western blotting and immunofluorescence. The levels of HIF-1α and NF-κB in nucleus, as well as the effect of HIF-1α or NF-κB inhibition on the expression of C3aR were also examined. We found that hypoxia induced upregulation of C3aR expression both in mRNA and protein level. The protein levels of HIF-1α and NF-κB in nucleus were increased in hypoxic condition. Pre-incubation with HIF-1α or NF-κB inhibitor, both inhibited the C3aR mRNA and protein expression induced by hypoxia. In addition, supplementation with HIF-1α inhibitor decreased the nuclear translocation of NF-κB. These results indicated that hypoxia could induce the expression of C3aR in tubular epithelial cells through HIF-1α/NF-κB pathway.]]> 2017/11/20 0:00:00 ZHAO Wen-Jin, CHEN De-Jun, LI Li-Juan and ZHENG Jing-Min 332true 2017/11/20 0:00:00 ZHANG Meng-Xia, LIU Qi, WU Hai-Yan, LI Xiang and TANG Sheng-Song 331true t test, confidence interval, etc. The accuracy can be as high as 98% through the machine learning method modeling, which is higher compared with the previous study. The KEGG (Kyoto Encyclopedia of Genes and Genomes) pathway analysis and GO (Gene Ontology) enrichment analysis indicated the significant correlation among eight and eighteen kinds of genes respectively. A functional analysis of the part of the eight pathways showed theirs close relationship at the level of gene regulation which indicted the identified signature genes play an important role in the pathogenesis of breast cancer and is very important for understanding the pathogenesis of breast cancer and the early diagnosis of breast cancer.]]> 2017/11/20 0:00:00 WEN Jian-Xin, WANG Xue-Dong, LI Xiao-Qin and CHANG Yu 330true 2018/6/21 0:00:00 FU Wei-Liang, TAO Wei and CHEN Fu-Yong 329true 2018/6/21 0:00:00 FANG You-Chen, ZHENG Yang, ZHANG Qing, LI Jing and XIE Xin 328true 2S was recognized recently as a new gaseous signaling molecule which plays beneficial roles in vasodilation and facilitation of long-term potentiation. The interaction of mammalian myoglobin (Mb) with H2S occurs via two distinct pathways: the first is the direct binding of H2S to the heme-Fe center of the Mb to form the Mb-H2S adduct, and the second is the covalent modification with H2S at a porphyrin vinyl group of the Mb heme to form the covalently modified sulfheme. We chose Mb as the prototype heme-containing protein and probed the interactions of different Mb mutants with H2S by UV-vis spectroscopy, and X-ray crystallography. The heme active sites of Mb mutants directly affect the binding of H2S to the proteins and alsothe formation of sulfheme derivatives. This provides critical experimental data for elucidating how heme-containing proteins modify the metabolism of endogenous H2S.]]> 2018/4/18 0:00:00 WU Yun-Xiang and YI Jun 327true n = 83) and double-on (DO, n = 34), the former type only discharged impulses to the onset of CF-FM sounds, and the latter type discharged impulses to the onset of both CF and FM components of CF-FM sound. The results also showed that whatever the bat processed the positive DSC signal or negative DSC signal, the 50%inter-pulse intervals(50% IPIs, i.e. 50% recovery time of response to echo)of SO and DO neurons were significantly shortened (P < 0.001) and concentrated in the short recovery region when the paired sounds were changed from the uncompensated condition to the best compensated condition. Moreover, the number of SO neurons whose rate of shortening of 50% IPI exceeded 70% was larger than that of DO neurons, and among those IC neurons which preferred positive DSC, the mean DSC selective range of SO neurons was also significantly wider than that of DO neurons. The above results suggested that SO neurons among IC neurons might make better use of bat's DSC behavior than DO neurons do to improve the response to echo so as to obtain the information of prey to the maximum and determine its relative velocity with the prey.]]> 2018/4/8 0:00:00 WANG Qiao-Chao, KONG Hui-Fang, CHENG Ya-Wen, WANG Hui-Mei, FU Zi-Ying, CHEN Qi-Cai and TANG Jia 326true N-FS36 is collected. High-resolution X-ray crystallography shows that FS36 interacts with Hsp90N at the ATP-binding pocket and this demonstrates that FS36 possibly substitutes nucleotides to bind to Hsp90N. The complex crystal structure and the interactions between FS36 and Hsp90N lay the foundation of the design and majorization of novel antitumor drugs.]]> 2018/4/18 0:00:00 LI Gen, LV Kai-Kai, LI Jian, YIN Xiu-Shan, ZHAO Dong and ZHANG Wan-Zhong 325true P < 0.05). Transfection of pcDNA3.1-MIR31HG significantly increased MIR31HG expression in ESCC cell lines(P < 0.01), inhibited cell viability, reduced S-phase cells numbers and increased the G1-phase cells numbers (P < 0.05). These results suggested that MIR31HG may reduce cell viability of ESCC cells though impeding cell cycle. Moreover, MIR31HG overexpression also significantly increased Caspase3 activity, Caspase3 and p53 expression and decreased Bcl-2 expression in ESCC cells (P < 0.05). Taken together, these results suggest that MIR31HG may hinder the development of esophageal cancer by inhibiting the proliferation of ESCC cells, which may provide a new strategy for the diagnosis and treatment of esophageal cancer.]]> 2018/6/27 0:00:00 CHEN Xiao-Qi, CHEN Xin-Ju, ZHANG Chuan-Lei, WANG Xin-Ting and YI Ai-Ying 324true 2018/5/21 0:00:00 ZHENG Jing-Min, CHEN De-Jun, YIN Guang, ZHAO Wen-Jin, LI Li-Juan and WANG Jian-Ping 323true Sox9 on male gonadal differentiation in Trachemys scripta was investigated through expression analysis and gene-modulating approach, providing important reference for the study of sexual differentiation and gonadal development mechanism of reptiles with TSD and an effective molecular techniques for controlling the sex of animals. Real-time PCR showed that the MPT-specific expression of Sox9 in embryonic gonads initiated early at stage 17, preceding the onset of gonadal differentiation, however, the FPT embryonic gonads exhibited extremely low expression level of Sox9 throughout the embryogenesis. Immunofluorescence further confirmed the SOX9 protein was abundantly expressed and mainly localized in the nuclei of precursor Sertoli cells in MPT gonads. The temperature shifts experiments showed that the expression of Sox9 in MPT→FPT (which was shifted at stage 16) gonads was significantly reduced from the stage 17, indicating that Sox9 can respond rapidly to the new temperature. Meanwhile, the expression of Sox9 was also down-regulated rapidly in the MPT gonads treated with estrogen. Loss-of-function analysis showed that 90.9% MPT embryos with Sox9 knockdown exhibited male to female sex reversal, characterized by obvious feminization gonads with well organized outer cortex and degradated medullary area. These above results indicate that Sox9 is required for testicular differentiation in a TSD system.]]> 2018/4/18 0:00:00 ZHANG Hai-Yan, SUN Wei, ZHOU Ying-Jie, BAO Hai-Sheng, CAI Han, QIAN Guo-Ying and GE Chu-Tian 322true 2018/5/18 0:00:00 LV Hui-Jie, ZHU Ze-Mei, CHEN Wei-Zhao, HE Jian-Qin, YANG Si-Si, ZHANG Kai-Fang, FENG Shui-Dong and LING Hong-Yan 321true 2018/4/10 0:00:00 LI Rui-Fang, LI Hong, GUO Chun-Yang and YANG Sa-Ru-La 320true 2018/3/27 0:00:00 CHEN Jun, JIANG Ting, LI Meng-Qi and ZHAO Guo-Jun 319true 2018/4/19 0:00:00 HUANG Lu, FENG Zhou-Yan and WANG Zhao-Xiang 318true 7, isocoumarins derivative 10 and Clavepictine analogue 11, were determined with low toxicity ultimately according to the predictive ADME simulation and predictive toxic simulation. Binding model analysis revealed that these 3 candidate compounds are all good drug-like PTP1B inhibitors, of which the PTP1B inhibitory activity of compound 10 and 11 haven’t been reported before, of which in vitro PTP1B enzyme inhibition of compound 10 was tested with IC50 values of (74.58±1.23) μmol/L, which is potential for the treatment of type Ⅱ diabete.]]> 2018/4/19 0:00:00 ZHANG Qian, GAN Qiang, LIU Xia, CHEN Xi and FENG Chang-Gen 317true 2018/3/21 0:00:00 FANG Long-Jie, ZHANG Xi-Cheng, ZHANG Cheng, ZUO Hao-Yi, ZHU Jian-Hua, GAO Fu-Hua and PANG Lin 316true 2018/3/21 0:00:00 KONG Zhen-Zhen, ZHAO Li-Ling, TANG Yan, WANG Ji-Hua and CAO Zan-Xia 315true σe) was so low as to make “cross-over” frequencies appear, meanwhile the frequency of the E-field f was adjusted in the range of the “cross-over” frequencies from Low-f0 to High-f0. Subsequently, three parameters affecting cell stretching were systemically discussed, and the results suggested that it was a prerequisite for cell electro-stretching that the conductivity of cell suspension (σe) was lower than that of cell cytoplasm (σc). In addition, increased voltage or decreased frequency of the E-field can also prompt cell stretching. Finally, the correlation between cell mechanical behavior and cell functional variation of NB4 cells was investigated via the optimized cell electro-stretching method, and the results showed that DOX treatment could make NB4 cells produce apoptosis and stiffen. Taken together, this study provides a novel cell electro-stretching method for the simple and efficient detection of cell deformability.]]> 2018/3/21 0:00:00 Bai Guohua, WANG Wen-Xi and TAN Qiu-Lin 314true 2018/1/16 0:00:00 LI Huai-Zhou, ZHOU Hai-Yan, YANG Yang, YANG Xiao-Jing, WANG Hai-Yuan and ZHONG Ning 313true Bartonella henselae secretes 7 Bep effector proteins (BepA-G) and injects these proteins into host cells using a type Ⅳ secretion system. Among these effector proteins, BepD-F contain multiple copies of Glu-Pro-Ile-Tyr-Ala (EPIYA) motifs in their N-terminal regions. After injection into host cells, the tyrosine residues in these EPIYA motifs will be phosphorylated by SFK, and the tyrosine-phosphorylated EPIYA motifs can subsequently interact with various SH2 domain-containing proteins from host cell and interfere with the SH2 signaling pathways of the host cells. In addition to B. henselae Bep effector proteins, numerous pathogenic bacteria effector proteins had been identified previously to contain EPIYA motifs. They were also found to utilize their EPIYA motifs to disrupt the SH2-signaling pathways of respective host cells, although they share no obvious overall sequence similarities. In contrast, only 6 proteins containing EPIYA motifs were found in human proteome, much lower than random occurrences, probably due to its toxicity in human cells. JAM-A is one of EPIYA motif containing proteins occurring in blood platelet, which can recruit Csk to avoid thrombus formation. BepE was reported to interact with Csk and disturb Csk-related signaling pathways. Here we reported the crystal structures of complexes between Csk and the phosphorylated EPIYA motifs from BepE and JAM-A, and biophysical measurements of their binding affinities. The complex structures show that Csk binds with two phosphopeptides in a similar way. The Csk SH2 domain binds with phosphotyrosine. Phosphopeptides are perpendicular to β-sheets in SH2 domain. Results of SPR show that the affinity of BepE with Csk is much higher than that of JAM-A. These results suggest that the EPIYA motif of toxic proteins binds with the SH2 domain of human Csk by a higher affinity, then disturb the SH2 domain proteins involved signaling pathways. The results reported here laid a solid foundation for our understanding of the structural basis of how toxic EPIYA motifs interrupt the SH2 signaling pathways of host cells.]]> 2018/1/16 0:00:00 ZHANG Ke, LIU Qing-Bo, HU Yong-Lin and WANG Da-Cheng 312true 2018/1/16 0:00:00 CHEN Yi-Ting, ZHANG Feng, ZHAO Jia, YU Jia-Feng, SHA Yu-Jie and WANG Ji-Hua 311true -3% and 61.9%, respectively. BMP-2-PLLAms/FGF-2-nHA/PLGA composite scaffolds were obtained with the pore size of 100-200 μm, the porosity of 75.8%, the compressive strength of 6.8 MPa, and the degradation rate of 19.9% at 8 weeks. The cumulative releases of FGF-2 and BMP-2 from the scaffolds were respectively about 77.1% and 44.2% after 7 days, 84.9% and 61.5% after 14 days. The results of BMSCs proliferation and differentiation showed that FGF-2 and BMP-2 released from the composite scaffolds could significantly promote cell proliferation and differentiation, and present a high bioactivity. BMP-2-PLLAms/FGF-2-nHA/PLGA scaffolds realized the sequential delivery of BMP-2 and FGF-2, and could induce BMSCs proliferation and differentiation effectively.]]> 2018/1/16 0:00:00 BAI Yan, BAI Li-Juan, CHEN Hua-Li and ZHOU Jing 310true hdac3 in CD4+CD8+ double positive(DP) stage of thymocytes using the cd4-cre transgene .The CD4Cre-mediated hdac3 deletion did not impact T cell development in the thymus but resulted in a dramatic loss of peripheral T cells. In addition, peripheral T cells in hdac3 knock-out mice showed a dominant activation/effector/memory phenotype. Mechanism analysis revealed an increased cell apoptosis which was accompanied by an accelerated cell proliferation in the peripheral T cells of hdac3 knock-out mice. Moreover, Fas and FasL positive cells and FasL expression increased significantly in the peripheral T cells of hdac3 knock-out mice. In vitro TCR activation did not affect the apoptosis of normal peripheral T cells, but dramatically increased apoptosis of peripheral T cells from hdac3 knock-out mice. Our results presented here indicate an important role of HDAC3 in maintaining homeostasis of peripheral T cells by refraining them from activation- induced cell death.]]> 2018/1/16 0:00:00 WANG Shan, TIAN Feng, QIAN Ye, LIU Ying, LI Hui-Ting, ZHANG Ai-Hong, HOU Zhi-Hong, LIU Ya-Nan, LI Juan, ZHANG Yan-Shu, ZHAO Yong and ZHENG Quan-Hui 309true Oryza sativa ssp. japonica cv. Nipponbare) at 24 h after treatment with 50 μmol/L Cd, was identified in cadmium-responsive transcriptome profiles analyzed by RNA-Seq. In this study, we isolated OsPDR from rice (Oryza sativa cv. Nipponbare) and the metal ions transport activity of OsPDR was analyzed. The results of metal tolerance experiments showed that overexpression of OsPDR can enhance yeast’s tolerance to Co, but not to Zn, Ni or Cd, and higher accumulation of Co was observed compared to the empty vector-transformed yeast as determined by inductively coupled plasma mass spectrometry (ICP-MS). The EGFP-OsPDR fusion protein was localized to the vacuolar membrane using confocal microscopy. These data suggested that OsPDR might play an important role in Co homeostasis. Additional studies are needed to understand the function of OsPDR in plants.]]> 2018/11/1 0:00:00 TIAN Si-Qi, QIAO Kun, WANG Fan-Hong, LIANG Shuang, WANG Hong and CHAI Tuan-Yao 308true Chlamydomonas reinhardtii under nitrogen depletion condition is a model system to investigate the pathway of lipid biosynthesis. Substantial amount of data have been accumulated using “omics” approaches recently, however, the identification of reference and biomarker proteins in nitrogen depletion-mediated triacylglycerol biosynthesis in C. reinhardtii is limited. In this study, C. reinhardtii CC-124 grown in control and nitrogen depleted medium were surveyed to compare the morphology, cell density, lipid content, and total protein content at multiple time points (0, 1, 2, 4 and 6 days). Under nitrogen depletion treatment, the culture turned yellow from green and the A750 and cell number was decreased, indicated that the cell growth was retarted. Furthermore, the concentration of neutral lipid was up-regulated based on nile red staining assay, while the coomassie brilliant blue staining assay indicated that the concentration of total protein was decreased. Western blot (WB) was performed to detect the expression patterns for 20 candidate C. reinhardtii proteins at different time points. To identify the reference proteins under nitrogen depleted condition, the Pearson’s correlation coefficient (PCC) between the content of total proteins and candidate proteins was calculated. Three proteins (Histone H3, RBCL (ribulose-1, 5-bisphosphate carboxylase/oxygenase large subunit) and BCR1 (biotin carboxylase, ACCase complex 1)) were selected due to their significant positive correlations (P < 0.01) in both nitrogen depleted and control conditions. Comparison of average relative fold change (ARF) of the candidate proteins indicated that the top 3 proteins are ATPs-β (ATP synthase CF1 beta subunit), GAP2 (glyceraldehyde 3-phosphate dehydroase 2) and RMT1 (rubisco large subunit N-methyltransferase 1), their ARFs are 180.59, 52.90 and 12.48, respectively, which were then chosen as biomarkers. Futhermore, to compare the earliest time point at which biomarker band can be detectable, protein samples at early stage (0, 2, 4, 8, 12, 18, 24 and 48 h) at nitrogen depletion treatment were collected and analysed by WB, it was showed that the induction bands of ATPs-β, GAP2 and RMT1 appeared at 8, 18 and 12 h, respectively. Thus, ATPs-β was the most appreciate biomarker since it is the earliest showed and the largest degree varied among candidates proteins tested. The reference and biomarker proteins identified in this study will provide help for the mechanism investigation of nitrogen depletion responses and lipid biosynthesis in C. reinhardtii, the expression profiling acculumated for candidate proteins can be refered by the reseach community.]]> 2018/12/21 0:00:00 SHI Jia-Nan, DU Tie-Min, CHEN Yue, ZHOU Yan, YANG Ya-Ru, LI Li-Yun, DOU Shi-Juan, LIU Li-Juan and LIU Guo-Zhen 307true 3+-Catechin complex (Yb3+-C) were systematically studied in this work. The experimental results showed that the optimum coordination mole ratio of Yb3+-C was 1∶4. The minimum inhibitory concentration (MIC) of Yb3+-C was significantly lower than that of single Yb3+ and C. Yb3+-C exhibited high antibacterial activity against Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa and Salmonella, and more effective for Staphylococcus aureus and the MIC value was only 0.0963 mmol/L. Further investigation showed that the sterilization time of Yb3+-C was greatly reduced to 2 h. In addition, the analysis of scanning electron micrograph (SEM), transmission electron microscope (TEM), and adsorption and distribution of Yb3+ in cell membranes and intracellular indicated the stronger affinity and penetration ability of Yb3+-C to bacterial cells. Therefore, the Yb3+ in the form of Yb3+-C is more easily accumulated to the effective concentration in the cytoplasm, resulting the damage of cellular structure and apoptosis.]]> 2018/11/19 0:00:00 YU Kang, LIU Lu, LI Ke, YANG Xi, LI Xia, LIAO Xue-Pin and SHI Bi 306true Rad9 gene, knockdown of Rad9 expression or removal of Rad9 protein led to inefficient DNA end-joining repair. Furthermore, loss of Rad9 impaired the DNA damage-induced binding of Ku70 to the chromatin and attenuated the DNA damage-induced kinase activity of DNA-PKcs. Taken together, our data unveil a novel functional interplay between Rad9 and the NHEJ protein Ku70, indicating a role of Rad9 in NHEJ repair through modulating the activation of the DNA-PKcs/Ku70/Ku80 complex.]]> 2018/10/22 0:00:00 FAN Ying-Jun, LIU Yu-Heng, WANG Yu-Lan, KONG Bing-Jie, ZHAO Yun, YE Chen, AN Li-Li and HANG Hai-Ying 305true Ralstonia solanacearum, a soil-borne destructive plant pathogen, has an unusually wide host range, and causes a bacterial wilt that seriously affects the production of many economically important crops in the world. Thus, investigation of physiological metabolism in R. solanacearum will be helpful to develop new ways to control the bacterial wilt. Fatty acids are compulsory components of bacteria. However, the fatty acid biosynthestic mechanism is still unclear in R. solanacearum. In this paper, we identified acyl-CoA desaturase and cyclopropane fatty acid synthase of R. solanacearum GMI1000, and characterized their functions in biosynthesis of unsaturated fatty acid or cyclopropane fatty acid. First, R. solanacearum RSc2450 (desA) complemented Shewanella oneidensis desA fabA mutant growth on LB without supplemented oleic acid and caused E. coli fabA mutant to produce unsaturated fatty acid. Furthermore, deletion of desA caused R. solanacearum grows weak on BG plate and reduces the palmitoleic acid production. The results showed though R. solanacearum RSc2450 encodes an acyl-CoA desaturase and involves in unsaturated fatty acid biosynthesis, R. solanacearum might possess a novel unsaturated fatty acid biosynthetic pathway. Next, of the two putative cyclopropane fatty acid synthases encoded genes in R. solanacearum, only cfa1 (RSc0776) restored E. coli cfa mutant YYC1257 growth in low pH medium and to produce cis-9, 10-methylene palmitic acid. And deletion cfa1 mutant was sensitive to low pH and high osmotic pressure, and lost the ability to produce cis-9, 10-methylene palmitic acid. These indicated that cfa1 involves in cyclopropane fatty acid synthesis in R. solanacearum and plays roles in adaption to low pH and high osmotic pressure. Moreover, Cfa2 (RSp1446) does not have cyclopropane fatty acid synthase activity, and its function needs further study.]]> 2018/10/22 0:00:00 DONG Hui-Juan, FAN Zhi-Yong, KUANG Cheng-Wei, LI Xian-Qi and WANG Hai-Hong 304true Xenopus. However, as Tiki1 gene is absent in rodents such as mice, it is impossible to use mice or rat to study its role in mammals. In this study, we generated Tiki1 gene modified pigs using CRISPR/Cas9 system combined with somatic cell cloning technology to study the role of Tiki1 gene in pig development. Aligned the human Tiki1 mRNA sequence provided by professor He Xi"s team with the pig genome database, we selected two target sites (g1 and g2) with the top 2 highest sequence identity at the predicted pig Tiki1 gene locus. The sgrRNA plasmid was constructed to transfect porcine fetal fibroblasts, and 52 single cell clones were screened and sequenced. We finally selected 5 single-cell clones with biallelic knockout mutations at target site g1 and 3 single-cell clones with biallelic knockout mutations at target site g2 as nuclear donors for constructing Tiki1 knockout pigs. A total of 720 recombinant embryos were constructed and transferred into three surrogate sows and one of them was successfully pregnant monitored by B ultrasound. A total of 13 cloned piglets (ten living piglets and 3 dead piglets ) were produced, and 12 of them were biallelic knockout mutations at Tiki1 locus. Both the living and dead Tiki1 gene knockout cloned piglets were developed normally and the living piglets have survived healthy till now. The results indicate that the role of Tiki1 gene on early development of pigs is different from that of frogs. The specific role of Tiki1 gene in the early development of pigs needs to be further investigated.]]> 2019/12/20 16:05:25 WU Cai-Xia,LIU Zhao-Ming,YAN Quan-Mei,ZHANG Quan-Jun,ZHAO Yu,OUYANG Zhen,FAN Na-Na and LAI Liang-Xue 303true in vivo. These results demonstrate that Ei24 is required for the development and function of iNKT cells.]]> 2019/12/20 16:04:25 DU Zhi-Wei,ZHANG Yu-Wei,YUAN Lin,BAI Li and ZHANG Hui-Min 302true 2019/11/5 15:25:15 HU Han-Han,FENG Zhou-Yan,WANG Zhao-Xiang,ZHENG Lv-Piao and HUANG Lu 301true 2019/11/5 15:45:00 ZHANG Jun-Wen,CUI Ying-Bin,CHEN Hong,HUANG Bing-Ren and LIU Fu-Sheng 300true 2019/11/5 15:55:52 ZHANG Wei-Wei,GAO Huan-Yu,SUN Qing,LIANG Kai,XIE When-Chun and LI Yan 299true 2019/9/25 0:00:00 WANG Xue-Dong,SHANG Wen-Hui,LI Xiao-Qin and CHANG Yu 298true PLK1, CCNB1, CCNA2 and CDC20, and promoted cell cycle progression. PLK1 protein, found elevated in Centlein knock out cells, interacted with Centlein in vivo. We propose that centrosomal PLK1 exerting control over the cell cycle relies upon the interaction with Centlein.]]> 2019/9/25 0:00:00 YANG Ling,ZHANG Ying and YUAN Li 297true Polygonum cuspidatum Sieb. et Zucc., a traditional Chinese herb used for the treatment of cough, suppurative conditions, and hypertension. The Pc2PS shared 54%–56% amino acid sequence identity with 2PSs of other species. Enzymatic activity analysis showed that Pc2PS could catalyze the condensation of one molecule of acetyl-CoA with two molecules of malonyl-CoA at almost the same efficiency as that of three molecules of malonyl-CoA alone for production of 4-hydroxy-6-methyl-2-pyrone. Thus, the presence or absence of acetyl-CoA in the reaction mixture did not affect catalytic efficiency. This catalytic property of Pc2PS differs from that of previously reported Gh2PSs, which catalyzed malonyl-CoA condensation to form the same products as Pc2PS but at a lower rate in the absence of acetyl-CoA. In addition, we determined the kinetics of Pc2PS with malonyl-CoA alone as the substrate for the first time. Pc2PS showed tissue-specific expression in P. cuspidatum; Pc2PS was predominantly expressed in the root, whereas few transcripts were detected in the leaf. This study enriches the known diversity of 2PS proteins, and provides a novel genetic resource for 2-pyrone biosynthesis.]]> 2019/9/25 0:00:00 CHEN Mo,WU Zhi-Jun,WANG Xiao-Wei,HU Hong-Yan,CAO Jie,CHAI Tuan-Yao and WANG Hong 296true S.typhimurium, the biofilms were detected by crystal violet staining method, biofilm integrity, biofilm biomass, bacterial activity, and the intracellular ROS levels of S.typhimurium S025. Our results showed that, after treatment with iron oxide (Fe3O4) nanozyme and iron sulfide (nFeS) nanozyme during biofilm formation for 48 h, the A550 value, biofilm height, biofilm biomass, and bacteria viability were significantly decreased compared to the untreated group, but the intracellular ROS levels remarkably increased. Moreover, to determine whether there is a similar inhibitory effect on pre-formed biofilms, Fe3O4 nanozyme and nFeS nanozyme were incubated from the apical side of the biofilms for different time. As expected, both nanozymes appreciably destroyed the biofilms that had formed. Importantly, nFeS nanozyme showed better efficacy than Fe3O4 nanozyme in above biofilm treatment. Taken together, these findings clearly demonstrated that iron-based nanozymes, as a suitable agent, regulated intracellular ROS levels to prevent the biofilm formation of S.typhimurium and destroyed the matured biofilm, which is helpful to preventing and treating relevant diseases caused by biofilm of S.typhimurium.]]> 2019/7/1 11:05:29 YIN Yin-Yan,WU Meng-Xing,QIN Tao,HE Kai-Ming,XU Nuo,SHI Yi,LIU Qi,PENG Da-Xin and GAO Li-Zeng 295true 2019/7/1 11:09:07 ZHANG Chan-Juan,DU Ke,AO Bao-Xue,ZHU Neng,YAN Tao,XIE Zhie-Zhong,LIAO Duan-Fang and QIN Li 294true 2019/7/1 11:13:30 LI Duo-Fang,GU Jian-Zhong,GENG Jin-Peng,CAO Tian-Guang,ZHAN Yong,SU Xiao-Jing and TANG Cheng-Chun 293true AC3 deletion. DNA methylation plays a key role in animal development and regulation of gene expression. In the present study, whether the DNA methylation level of gene promoter, as well as their associations with the expression of the genes in MOE will be altered after AC3 deletion, was investigated by using methylated DNA immunoprecipitation chip (MeDIP-chip), methylation-specific PCR (MSP) and real-time fluorescence quantification PCR. The data showed that the DNA methylation levels of promoters of 1 978 genes were altered in AC3-deficient mice, accounting for 9% of the total number of genes. Of which 727 genes with their promoter’s DNA methylation levels were elevated, 1 251 genes with their promoter’s methylation levels were lowered. The functions of these genes are mainly involved with olfactory receptor, neurodevelopmental, cAMP signaling pathway, ATP-binding, calcium regulation, acetylation modification, and transcription factors. It was further confirmed by MSP that methylation levels of promoter of the olfactory receptor genes Olfr1153, Olfr231, Olfr378, Olfr651 and Olfr691 were increased, whereas methylation level of the promoters of Cngb1, Pde4a and Olfr1394 were decreased. In line with MSP results, qRT-PCR data showed that the expression levels of Cngb1, Hcn4, Olfm1, Olfr1394, Olfr1153, Olfr231, Olfr378 and Olfr691 were significantly decreased, whereas the expression levels of Pde4a and Olfr651 were significantly increased. In conclusion, the methylation levels of promoters of olfactory receptor genes, neurodevelopmental related genes and cAMP signaling pathways in MOE are modified significantly after AC3 deletion, which affected the transduction of signal pathways such as nucleotide excision and repair, DNA replication and mismatch repair, thus comprehensively regulating the number and level of gene expression in MOE of mice.]]> 2019/5/22 0:00:00 ZHOU Yan-Fen,WANG Ya-Wen,WANG Xiao-Ting,SHU Li,LI Shu-Juan and WANG Zhen-Shan 292true in vitro culture. In this study, the effect of glycosphingolipid GM1 on the contact inhibition of human mammary epithelial cell lines MCF-10A, human breast cancer cell lines BT-549 and SK-BR-3 were investigated. Changes of cell proliferation of MCF-10A, BT-549 and SK-BR-3 cells at low and high cell density was explored. Expression of GM1 in different cell density was detected by flow cytometry. Exogenous GM1 at different cell density was added to explore the effect on cell proliferation. B3GALT4, the GM1 synthase, was knocked down or overexpressed in BT-549 and SK-BR-3 cells by using lentiviral vectors. Then, proliferating ability were tested by cell counting and related pathways was assayed by Western blot, in stable-transfected cell lines. Results showed cell proliferation was inhibited and GM1 expression increased at low and high cell density compared with normal density. Exogenous addition of GM1 at low and high density cells inhibited cell growth, but it had no influence on cell growth at normal density. Down regulation of GM1 promoted cell proliferation at low and high cell density, and overexpression of GM1 had the opposite effect. Together, these results indicate that GM1 inhibit MCF-10A, BT-549 and SK-BR-3 cells proliferation at low and high cell density in vitro, which may potentially provide the experimental basis for further research on its molecular mechanisms.]]> 2019/5/22 0:00:00 ZHUO Ding-Hao and GUAN Feng 291true 3D dataset showed that the prediction accuracy of donor and acceptor sites were 93.39% and 90.46% respectively, obviously higher than that of the compared methods. The positional features based on the chi-square statistical difference table can effectively characterize DNA sequences, and have application prospects in signal site recognition of molecular sequences.]]> 2019/5/22 0:00:00 ZENG Ying,CHEN Yuan and YUAN Zhe-Ming 290true P<0.001), TNF-α (P<0.001) and IL-6 (P<0.001) in cell culture supernatants. Simultaneously, SCFAs mix inhibited the increase of iNOS, TNF-α, IL-6 (P<0.001) and inflammasome NLRP3 (P<0.001) mRNA in BV-2 cells induced by LPS. Further study showed that SCFAs mix could inhibit LPS-induced high expression of TLR4, MyD88, TRAF6 and NF-κB proteins, which were the key molecules of the inflammatory signaling pathway in BV-2 cells. In conclusion, proper concentration of SCFAs mix could function as a protective role to inhibit LPS-induced microglial inflammatory responses by regulation of the TLR4/MyD88/TRAF6/NF-κB signaling pathway.]]> 2019/5/22 10:23:39 JIA Xue-Bing,ZHOU Zhi-Lan,ZHANG Bo-Ping,ZHAO Li-Ping,QIAO Chen-Meng,SUN Meng-Fei,SUN Xiu-Lan,CHEN Yong-Quan,CUI Chun and SHEN Yan-Qin 289true P<0.05); 2-DG decreases the ATP level of MCF-7 and MCF-7-M cells, and the effect of lowering the ATP level of MCF-7-M cells is more obvious (P<0.01). The glucose uptake and lactate secretion of MCF-7-M cells were significantly higher than those of MCF-7 cells (P<0.01). After the treatment with API-2, the glucose consumption and lactate secretion of MCF-7 and MCF-7-M cells were significantly reduced (P<0.01). The expressions of GLUT-1, HK2 and PKM2 in MCF-7-M cells were significantly higher than those in MCF-7 cells (P<0.01). Both LY294002 and API-2 inhibited the expression of GLUT-1 in MCF-7-M cells (P<0.05). After the treatment with 3-BrPA, the drug sensitivity of MCF-7-M and MCF-7 cells to 5- FU was significantly enhanced (P<0.01). In conclusion, cytoplasmic M-CSF activates glycolysis by induce GLUT-1, HK2 and PKM2 protein expression in MCF-7 cells; PI3K/AKT signaling involves the pathway that glycolysis was activated by cytoplasmic M-CSF in MCF-7 cells.]]> 2019/4/19 0:00:00 TIAN Yun,LIU Zhen,NING Qian,MO Zhong-Cheng,ZHANG Meng-Xia and TANG Sheng-Song 288true 2019/4/19 14:56:36 WANG Qi,DING Yan-Shu,GENG Bao-Bao and NIE Yu-Min 287true V (maximum volume change), AOD(accumulative osmotic damage) and J (accumulative chemical damage) were used to evaluate the damage of oocyte. A comprehensive parameter s, which based on the traditional parameters and the information entropy theory，was proposed to evaluate the damage. Oocytes loading experiments were conducted to verify the effectiveness of s. The results showed differences from the results of traditional damage assessment parameters, which cannot come to the unified conclusion. By analyzing the relationship between blastocyst rate and s, we found that there is a highly negative correlation between s and blastocysts rate. That indicates that the comprehensive damage assessment parameter s can be used to assess cell damage and breaks a new path for cell damage evaluation.]]> 2019/3/25 14:58:29 SHAO Wen-Qi,GUO Ying-Ying,DAI Jian-Jun,ZHANG De-Fu and ZHOU Xin-Li 286true E.coli SUMO expression system, and here we report the structural envelope of the C terminal domain of gp16 obtained through small angle X-ray scattering (SAXS) technology. The gp16 C terminal domain gene was recombinantly expressed using the SUMO tag. The gp16 gene was flanked by an N-terminal 6His-SUMO tag and purified by Ni-NTA affinity chromatography. High purity target protein monomer was obtained by size exclusion chromatography. This brings the purity of the target protein to about 95%. Then the concentration of the target protein to 1.3 g/L, and the SAXS data were collected at Shanghai Synchrotron Radiation Facility (SSRF) beamline BL19U. The resulting molecular envelop highly resembles the homologous protein FtsK, and the curves calculated by the CRYSOL software are highly coincident with the experimental curves. Through recombinant expression, purification and structure analysis, we obtained the solution structure of the gp16. Structural information derived from this study laid the foundation for future structural and functional research of gp16 C terminal, ultimately targeting the viral mechanisms for infection.]]> 2019/3/25 15:02:46 WU Fei-Fei,CAI Ru-Jie,CHEN Ting,ZHANG Yun-Long and LU Chang-Rui 285true -/- mice exhibited a phenotype of glycometabolic dysregulation, such as higher fasting blood glucose level, accumulation of hepatic glycogen, recession of blood glucose profiles, and elevation of gluconeogenesis. The expression levels of PEPCK were remarkably up-regulated in the liver tissues of HBXIP-/- mice. Then, we validated that HBXIP expressions were negatively correlated with those of PEPCK in 30 clinical liver tissues. Mechanistically, luciferase reporter gene assays and ChIP assays showed that HBXIP could inhibit the expression PEPCK at transcription level. Taken together, our findings indicate that HBXIP restrains hepatic gluconeogenesis through suppressing the expression of PEPCK.]]> 2019/2/20 0:00:00 SHI Hui,FANG Run-Ping,ZHANG Wei-Ying,LI Ying-Hui,SI Chuan-Ping,XIONG Hua-Bao and YE Li-Hong 284true 2019/2/20 11:40:55 YUE Hai-Yan,QIN Jing,WANG Wen-Song,YANG Ye-Ning,YI Lan,WANG Juan,TANG Yu-Xian,HE Jie,SU Qi and TAN Hui 283true 2019/1/18 0:00:00 FENG Lei,JIANG Li,LI Shan-Fei,ZHANG Jian,LIU Hai-Bo,JI An-Quan,YE Jian,WANG Gui-Qiang and LI Cai-Xia 282true 2020/4/20 0:00:00 SUN Xiang-Yu,WANG Yong,LIAO Ran,SUN Shu-Qing and MA Hui 281true 2020/4/23 0:00:00 ZHAO Cheng-Hui,HONG Hao,LI Wan-Ying,LI Rui-Jiang,JIANG Shuai,LI Hao,CHEN He-Bing and BO Xiao-Chen 280true in vitro. The catalytic efficiency of AKR1Cs was detected by spectroscopic methods, and the effect of inhibitors on the catalytic reduction of 4-OHA by AKR1Cs was verified. It was found that four AKR1C isoforms can reduce keto-groups and double bonds of 4-OHA. AKR1C4 can rapidly catalyze the structural changes in 4-OHA with a conversion rate of almost 100%, followed by AKR1C3 and AKR1C1 with a conversion efficiency of about 30%. AKR1C2 has the lowest activity towards of 4-OHA, and the conversion efficiency is only about 20%. At the same time, the inhibitor showed a significant dose-effect relationship with AKR1Cs. Non-linear regression analysis showed that the inhibitors had a strong affinity for AKR1C3 and AKR1C4 with IC50 values of 47.4 μmol/L and 54.68 μmol/L, respectively. The inhibition of AKR1C1 and AKR1C2 is relatively weak, and the IC50 values are 77.37 μmol/L and 82.24 μmol/L, respectively. The above results indicate that 4-OHA can be rapidly metabolized by AKR1C4 which is expressed in the liver only which may contribute to the fact that 4-OHA and its many conjugated metabolites is not very effective after oral administration. Our data support the advantage of parenteral administration of the drug as a depot-formulation or as a preparation for transdermal delivery and also provides a new idea for further research on nano-drug carriers and derivatives of the drug in the future.]]> 2020/4/23 0:00:00 KONG Xi,TAO Si-Wen,Frank Heinrich WIELAND,YANG You-Zhe,Alexander Tobias TEICHMANN,MA Gen,CHEN Mu-Lan,XIE He-Song,WANG Hai,WANG Gui-Lin and WAN Run-Lan 279true Bacillus subtilis (BsuDnaG) was constructed from the data of X-ray small angle scattering (SAXS) analysis. The BsuDnaG is in extended state in solution. On the other hand, the ZBD and HBD domains could exhibit continuous conformational changes relative to the RPD domain. This study suggests the domains rearrangement in DnaG primase may facilitate its function in DNA replication.]]> 2020/4/17 0:00:00 LUO Hao,LIU Wen-Lin,ZHOU Ying-Qin,TAO Mei,LIU Zhong-Chuan and WANG Gang-Gang 278true 2020/4/13 0:00:00 LI Bing-Yao,ZHANG Yan-Ting,LIN Qiu-Ping,WANG Lei,LI Si-Na,HAN Ding-An,ZHONG Jun-Ping,WANG Ming-Yi and ZENG Ya-Guang 277true in Arabidopsis was systematically investigated. It’s helpful for elucidating gene expression mechanism in plant genome.]]> 2020/4/13 0:00:00 XING Yong-Qiang,HE Ze-Xue,LIU Guo-Qing and CAI Lu 276true 2020/3/4 10:43:06 YANG Lin,GUO Shuai,MA Xiao-Liang,HOU Cheng-Yu,SHI Li-Ping,LI Jia-Cheng and HE Xiao-Dong 275true Pseudomonas putida has important application values in environmental pollution bioremediation and industrial polyhydroxy fatty acid (PHA) production. Bioinformatics analysis showed Pseudomonas putida genome encodes six FabG homologues with E. coli FabG, of which PpFabG5 shows the highest sequence similarity (76.5%) and other five PpFabGs also have high similarity (about 50%). Except PpFabG4, the other homologs have the conserved catalytic activity sites and N-terminal cofactor binding sites. So the paper used different methods, including the genetic complementary, catalytic activity analysis in vitro, gene deletion in vivo, and mutant characteristic analysis, to study the biological functions of the six homologs. The results showed that only PpfabG1, PpfabG3 and PpfabG5 could restore the growth of E.coli fabG temperature-sensitive mutant CL104 at 42℃, and the complementary strain of PpfabG1 grew weakly. While in vitro analysis, PpFabG1, PpFabG3 and PpFabG5 also showed the catalytic activities in the initial and extension reactions of fatty acid synthesis, although the activity of PpFabG1 was really weak. PpFabG6 only had catalytic activity in the initial reaction. PpfabG5 is an essential gene for growth, which can’t be deleted in the genome. While the deletion of the other PpfabG individually does not affect bacteria growth or the compositions of fatty acids comparing with wild type strain. However, the motility of PpfabG1 or PpfabG3 deletion mutant decreased, PpfabG3 or PpfabG6 deletion affected the biofilm formation, and PpfabG3 or PpfabG6 deletion mutant showed higher tolerance to H2O2. These results proved different FabG homologs contained different biological functions, especially when countered with stressful conditions.]]> 2020/3/4 10:51:36 GUO Jian-Ying,CHEN Bo,LI Xian-Qi,KUANG Cheng-Wei,WANG Hai-Hong,MA Jian-Rong and YU Yong-Hong 274true 2020/3/4 11:00:49 YANG Yan,SUN Yu-Ling,LIU Pei,QIAN Lin-Nan,GUAN Dong-Lang,JIAN Tun-Yu,ZHU Chan,YU Guang,WANG Chang-Ming,ZHU Jing,LI De-Feng and TANG Zong-Xiang 273true 2020/12/21 10:14:48 HAN Jun-Ping,ZHAO Lei,WANG Qing-Guo,JIANG Li,XIE He-Xin,ZHUANG Bin,ZHAO Li-Jian,LI Tang-Song,WU Jin and LI Cai-Xia 272true 2020/12/21 0:00:00 YU Song-lin,YU Tao,ZUO Zhen-tao,WANG Jing,XU Zong-sheng,YANG Kai-yuan,REN Xiao-hui,ZHAO Yuan-li and LIN Song 271true 2020/12/21 0:00:00 WANG Xiao-Yan,QIN Ya-Ling,HAN Qun,GU Xi-En,YAN Zi,FU Kui,LI De-Feng and DENG Kai 270true 2020/12/21 0:00:00 CHEN Ni,QIN Yu-Rong,GAO Huan,LI Yu-Qin and SUN Peng-Fei 269true 2020/10/26 0:00:00 Pei Qing,Lin Yin,Rong Yi,Liu Meng-Xue,Wu Shi-Ping and Yang Xiao-Fei 268true 2020/10/26 0:00:00 LU Lin,LIU Yang and LI Chun-Hua 267true TM 2500, following with analyses including differential expression gene GO annotation and KEGG analysis. As a result, about 94.38% genome database were covered in the human reference genome and 2 242 transcripts were detected as predicted new protein-coding genes. Compared with NOA, 8 045 DEGs were up-regulated and 1 150 were down-regulated in OA1. For OA1 and OA2, there were 1 538 and 420 DEGs for down-regulation and up-regulation. For OA2 and OA3, there were 1 275 for up-regulation and 1 690 for down-regulation. The numbers of up-regulated (1 834) and down-regulated (1 853) DEGs between OA3 and OA4 were similar. KEGG enrichment analysis indicated that DEGs were mainly enriched in RNA degradation pathway, basal cell carcinoma pathway, cancer pathway, melanogenesis pathway and signaling pathways regulating pluripotency of stem cells. The expressions of PRM1, PRM2, TNP1, UBXN6, CXCL16, NUPR2, CCDC136 and CRISP2 genes showed an increasing trend with time-sequence specificity. In addition, different extents of gene fusions were found in 5 cases of azoospermia testicular tissue expressed genes. In summary, the number of DEGs and their function, classification and metabolic pathways were obtained with different degrees of azoospermia. The DEGs such as spermatogenesis and sperm motility were screened out, which enriched transcriptome information for azoospermia and laid a foundation for the research of testicular tissue related genes. It may provide theoretical basis for revealing the molecular mechanisms of spermatogenesis in azoospermia patients and promoting the development of male infertility research. Notably, gene fusion may be associated with azoospermia.]]> 2020/9/22 0:00:00 LUO Bin,Lü Zi-li,SHAN Xu-Dong,ZHAO Qing-Mei,DU Zhen-Zhen,ZHANG Xia,LI Jun-Jun and LIANG Xin 266true 2020/9/18 0:00:00 HU Ping-Dong,MO Wei-Chuan,FU Jing-Peng,LIU Ying and HE Rong-Qiao 265true chi-square test, et al.. All indicated that the risk score calculated by the model can effectively predict the risk degree of patients(P<0.000, hazard ratio=2.551, 95% confidence interval=1.751-3.717). 1-5 year survival rates of HCC patients in the low risk group had 20%-30% higher than in the high risk group. Through the clinical data analysis, it was found that the combined biomarkers have a better prognostic effect than other clinical indicators, and can also be used as an independent prognostic factor. Target genes of four miRNAs were predicted, including AGO2, FOXO1, ROCK2, RAP1B, CYLD, et al., and enriched in biological processes such as cell proliferation, migration, apoptosis and immune response.]]> 2020/8/11 0:00:00 MEN Jing-Rui,TAN Jian-Jun and SUN Hong-Liang 264true Drosophila melanogaster. Simulated microgravity experiments performed using the random positioning machine (RPM) and male Drosophila melanogaster, and the activity and sleep of the flies were monitored using the DAM system (Trikinetics, USA). First, flies were exposed to simulated microgravity for a short time (3 days) with DAM system monitoring. Next, after being exposed to simulated microgravity for a long time (10, 20, and 30 days), flies were removed to be monitored for 3 consecutive days under control conditions. Then, the flies after short-term treatment (3 days) with greater impact were sampled to study the effects of simulated microgravity on the major circadian clock genes period (per), timeless (tim), clock (clk), cycle (cyc), cryptochrome (cry), and ddc, pale, trh genes encoding dopa decarboxylase, tyrosine hydroxylase, tryptophan hydroxylase, that involved in synthesis of neurotransmitter dopamine (DA) and 5-hydroxy-tryptamine (5-HT), and the contents of DA and 5-HT. The results showed that during short-term treatment, total activity accounts and activity accounts in wake period both increased, and total sleep and number of sleep episode both decreased at night; after short-term treatment, the relative expression levels of tim, clk, cyc and cry increased, and the relative expression levels of ddc, pale and trh increased; after long-term treatment, overall effect on activity and sleep was slight. This study suggests that it is feasible to study the effects of microgravity on activity and sleep of Drosophila melanogaster by simulated microgravity, and valuable reference can be provided for aerospace medical research.]]> 2020/7/17 0:00:00 ZHANG Hong-Ying,LI Zhi-Hui,ZHANG Lu,WANG Ya-Hong,ZHANG Zi-Yan and CAI Peng 263true 4/cm2. In this study, we screened the conditions of the best Vero cell culture system and confirmed that the conditions meet the continuous and stable passage of Vero cells.]]> 2020/7/17 0:00:00 YE Hua-Yue,JING Min-Min,SHAO Cong-Wen and ZHU Feng-Cai 262true 2020/6/10 0:00:00 ZHANG Dai,ZUO Zhen-Tao,RAO Heng-Yi and FAN Yong 261true 2021/3/18 0:00:00 WANG Xiao-Feng,TAO Gang,XU Ning,WANG Peng,LI Zhao and ZHOU Jie 260true 2021/3/18 0:00:00 WANG Kang,WANG Lei,LIU Pan,CUI Zhen,FENG Rui,CHEN Rui-Hong,CHEN Zhong-Hao,ZHU Qian-Hui and WANG Nan 259true 2021/3/4 0:00:00 JIAO Peng-Cheng,JI Jiao-Jiao and ZHAO Dong-xu 258true 2021/3/4 0:00:00 CHANG Meng-Yuan,WEI Xiao-Xi and LU Qing 257true 2021/3/4 0:00:00 LIU Heng-Heng,SONG Yan-Zhuo,LI Ji-Qiang,DING Yan-Zhi and GE Bao-Sheng 256true 2 = 28.9%, AUC = 0.82). Age had the greatest impact on the model performance (R2 = 22.9%, AUC = 0.77), and the AUC reached the maximum when BMI and 68 SNPs were added cumulatively (about 0.89). This study indicates that MPB has strong genetic heterogeneity in European and East Asian populations. Selecting a subset of SNPs can achieve prediction accuracy close to that of the complete set. The prediction model constructed here may help understand the genetic mechanism, early diagnosis and prevention of MPB in East Asian populations.]]> 2021/1/19 0:00:00 pansiyu,zhaowenting,fengrui,liqiong,jingxiaoxi,gaoxingjian,chenyan,liyi,taoxianming,liutianzi and licaixia 255true 2021/1/19 0:00:00 ZhangYuanyue,WangShuqing,LiuYan,LiuYankun,LiYuhui and LiYufeng 254true S. thermophilus at 2.94 ? resolution. On the basis of the structure comparison, DltD was considered as the terminal acyltransferase of the dlt operon, and it belonged to the SGNH-like family. An stDltD active center, including four blocks and a catalytic triad, conservatively exists in various Gram-positive pathogens. In addition, structural analysis showed that the stDltD catalytic center formed a strong positively charged groove docking with a glycerolphosphate molecule. Combined with previous reports, an updated working model was proposed for cross-membrane D-alanylation mediated by the dlt operon. The structural evidence provides more implications to clarify the biological function of stDltD and the process of transmembrane acyl modification.]]> 2021/9/23 0:00:00 ZENG Qi,TIAN Li-Fei,LIU Yan-Ping,YAN Xiao-Xue and XU Wen-Qing 253true Sporosarcina pasteurii (S. pasteurii) is a kind of Gram-positive bacteria from soil. Various applications have been developed based on the efficient urease activity that can induce the precipitation of calcium carbonate. However, the metabolic mechanism related to biomineralization of S. pasteurii has not been clearly elucidated. Especially, there are few studies on the gene structure of urease, regulation mechanism of expression and associated metabolism, which play key roles in biomineralization. Nowadays, the uncontrollability and instability of biomineralization reactions in the applications of S. pasteurii root in the lack of research on urease metabolisms. Therefore, it is urgent to further reveal the gene information, expression regulation and related metabolism of urease in S. pasteurii.In this paper, we compared the growth and gene expression of S. pasteurii BNCC 337394 under four different culture conditions through high-throughput transcriptome analyses. The four medium conditions were: (1) control group with yeast extract of 20 g/L; (2) ammonium group with yeast extract of 20 g/L and ammonium chloride of 10 g/L; (3) urea group with yeast extract of 20 g/L and urea of 5.62 g/L; (4) ammonium Tris group with yeast extract of 20 g/L, ammonium chloride of 10 g/L and Tris of 15.75 g/L. Transcriptome data were analyzed by bioinformatics methods of differential gene expression analysis, GO enrichment analysis, KEGG enrichment analysis and operon prediction.The results showed that there were significant differences in the growth of S. pasteurii among the four conditions. The bacteria could not grow and reproduce normally in the control group. There was a growth delay in the ammonium Tris group. The growth rate and trend of the ammonium group and the urea group with the same nitrogen content were similar. A total of 3 090 genes were generated and expressed in S. pasteurii in the experiment. There were 1 152 differentially expressed genes (DEGs) between the ammonium group and the control group, of which 411 were up-regulated and 741 were down-regulated. There were 1 362 DEGs between the urea group and the control group, of which 736 were up-regulated and 626 were down-regulated. There were 803 DEGs between the ammonium group and the urea group, of which 279 were up-regulated and 524 were down-regulated. There were 794 DEGs between the ammonium group and the ammonium Tris group, of which 281 were up-regulated and 513 were down-regulated. Furthermore, it was found that the expression of urease was significantly enhanced in the control group which was short of a nitrogen source comparing with the ammonium group and the urea group.GO and KEGG analyses revealed that the control group needed to enhance its basal metabolism and express more flagellum in order to survive. The pathways of electron transfer activity and oxidative phosphorylation were different in the ammonium group and the urea group. It indicated that the synthesis of ATP was associated with the hydrolysis of urea in S. pasteurii. Meanwhile, ammonium stimulated the expression of urease and ATP synthase in the ammonium group which was more significant than that of the urea group. The results of the ammonium Tris group indicated that a stable and moderate alkaline pH environment favored a high level expression of urease. Finally, the double operon gene structure of urease was predicted based on the two key characteristics of gene expression similarity and gene spacing.]]> 2021/9/23 0:00:00 PEI Di,LIU Zhi-Ming,HU Bi-Ru and WU Wen-Jian 252true 2021/9/23 0:00:00 QIU Fang,CHEN Chen,ZHANG Fang-Tong,MA Rui-Ya,SHI Li-Jun,SHENG Xin-Jun and LIU Xiao-Dong 251true t=60 s), ELF-EMFs were exposed to hippocampal neurons at 0, 0.09, 0.38, 0.76, 7.33 and 14.78 mT, respectively. Reactive oxygen species (ROS) and Ca2+ fluorescence response curves of hippocampal neurons were recorded in real-time, and the autocorrelation function between ELF-EMFs and fluorescence response was established. The results showed as follows: (1) The step properties of fluorescence response were clear at the burst times of ROS and Ca2+, which were important indicator to identify the real-time response; (2) The response time of ROS and Ca2+ to ELF-EMFs were delayed and inconsistent; (3) ROS and Ca2+ had dose-dependent response to ELF-EMFs; (4) The response of ROS to ELF-EMFs was complex; (5) The response of Ca2+ to ELF-EMFs was asymptotically stable. When correlation function exceeded 0.3, the real-time response of ELF-EMFs and ROS/Ca2+ was correlated. It was concluded that a real-time response method of intracellular ROS and Ca2+ to ELF-EMFs exposure was feasible for the evaluation of electromagnetic bioeffects.]]> 2021/9/23 0:00:00 CEN Ze-Nan,HUANG Wei-Wei,BAO Jia-Li,ZHENG Xiu-Xiu and ZHU Chao-Yang 250true in vitro experimental studies, such as lung cancer genetic screening and targeted drug screening. This article studied the vitrification method of living lung cancer tissue. Firstly, the needle immersion method was used to vitrify a single piece of lung cancer tissue, and the concentration and equilibrium time of the required cryoprotectant were optimized. Secondly, multiple lung cancer tissues were vitrified in cryotubes, and the volume of cryoprotectant solution and equilibrium time were optimized. Finally, the effects of the traditional slow freezing, rapid freezing without cryoprotectant, and vitrification methods were compared, and low-temperature microscope was used to analyze the damage mechanism of ice crystal. The results showed when 20% EG+20% DMSO+0.5 mol/L trehalose was used as the cryoprotectant, the equilibration solution and the vitrification solution are loaded for 3 min and 1 min, respectively, the needle immersion method and vitrification in a 0.25 ml cryotube have the highest tissue viability (79.96% and 80.44%) after recovery. Immunohistochemistry showed that the tissue structure of lung cancer after vitrification was less damaged, only a few positive expression of TUNEL in the cells, compared with slow freezing and fast freezing without cryoprotectants. Low-temperature microscopy showed that only a few small ice crystals appeared in and around the vitrified tissue, while obvious ice crystals appeared in slow freezing and rapid freezing.]]> 2021/9/23 0:00:00 LI Jia-Hui,DU Yu-Kun,YU Ke-Ke,XING Jie and ZHOU Xin-Li 249true HER2 genomic DNA reference materials (RM) and clinical samples, ddPCR and qPCR were used to study the commutability of HER2 genomic DNA RM, which can provide traceable RM with commutability for clinical laboratory testing. 29 clinical samples were randomly measured among the 5 levels of RM by real time quantitative PCR (qPCR) and droplet digital PCR (ddPCR). The averaged copy number ratio of HER2 to RPPH1 was calculated. According to the guideline EP30 of American Society for clinical laboratory standardization (CLSI) and the guideline WS/t356-2011 for Guideline for Evaluation of Matrix Effects and Commutability of the People’s Republic of China, a regression curve was drawn with the result of ddPCR as abscissa and the result of qPCR as ordinate, and the commutability of RM was evaluated by Deming regression method. If the test results of the proposed RM fall within the 95% confidence interval of its predicted, it is considered that the analyzed RM is communtable; if it falls outside the range, it is considered that the analyzed RM is not communtable. In addition, the results were compared with the ratio of copy number concentration when CEP17 gene was used as reference gene. HER2/RPPH1 of the five RMs determined by ddPCR and qPCR were: 1.91, 1.86; 5.70, 4.45; 16.94, 12.21; 22.38, 17.19; 35.38, 28.84, respectively, which fall in the prediction range, indicating the five RMs are communtable. Moreover, this was confirmed by the ratio of HER2/CEP17 determined by the two methods. However, the regression coefficient of HER2/RPPH1 (R2 = 0.97) was better than that of HER2/CEP17 (R2 = 0.78) and even one group of clinical sample fell outside the 95% confident interval. We used FISH to detect the cell lines used to prepare HER2 genomic DNA RM. The results showed that in the same nucleus, part of chromosome 17 showed HER2 amplification in the short arm, and part of chromosome 17 showed no HER2 amplification. Additionally, CEP17 was amplified in some cells, which indicates it is not suitable to be used as reference gene as this will cause false negative results. This confirms the necessity of using RPPH1 gene as reference gene when diagnostic of HER2. In conclusion, the five RMs are communtable, which can be used for the method validation and quality control in analyzing of HER2 copy number variation in clinical laboratories.]]> 2021/9/23 0:00:00 XING De-Chun,CHENG Bo,WANG Xia,GAO Ying,LIU Zheng,SUN Suo-Zhu,DONG Lian-Hua and YANG Jing-Ya 248true 2021/8/24 0:00:00 ZHANG Kang-Hui,LI Cheng-Shuang,DONG Lei,WANG Hui-Quan and ZHENG Yu 247true R2 between temperature and resistance of Ti/Pt temperature sensors of different thicknesses was greater than 0.999. Different cells trapped by this microchip were cultured on chip and monitored under a constant temperature environment of (37±0.015)℃. The temperature fluctuation range of human lung adenocarcinoma cell (H1975) (0.173℃) during metabolism was greater than that of hepatic stellate cell (HSC) (0.127℃). The average temperature of cancer cells H1975 (37.001℃) is higher than that of normal HSCs (36.989℃). In conclusion, this integrated microchip provides a tool of real-time monitoring cell temperature variation for the study of cell physiology and pathology.]]> 2021/8/24 0:00:00 ZHAO Xue-Fei,GAO Wan-Lei,YIN Jia-Wen,GUAN Yi-Hua and JIN Qing-Hui 246true P<0.01; P<0.05). At the same time, the transcription levels of LC3, ATG5 and Beclin-1 in oxidative stressed neurons were up-regulated (P<0.05; P<0.05; P<0.01). Correlation analysis showed that GALR2 was positively correlated with LC3, ATG5 and Beclin-1 (P<0.05; P<0.05; P<0.01). The treatment with M871, a specific inhibitor of GALR2, decreased the activity of hippocampal neurons under oxidative stress (P<0.01) , increased the number of autophagosomes (P<0.01) and transcription levels of LC3, Beclin-1 and ATG5(P<0.01) , and increased the ratio of LC3-Ⅱ/actin and P62 protein level (P<0.05) , showing that the autophagy of hippocampal neurons, which was up-regulated by oxidative stress, was inhibited with the inhibition of GALR2 expression, thus weakening the resistance to oxidative damage and decreasing the viability of neurons. Simultaneously, M871 treatment also decreased the up-regulated protein level (P<0.01) and the phosphorylation level of JNK (P<0.05) in oxidative stressed neurons, indicating that JNK is the downstream target enzyme of GALR2 in hippocampal neurons under oxidative stress. However, treatment with JNK specific inhibitor SP600125 lowered the ratio of LC3-II/actin, which was up-regulated by oxidative stress (P<0.01), showing the inhibition of JNK blocks the activation of the autophagic pathway by an up-regulated GALR2 in neurons. To sum up: under oxidative stress, the up-regulated GALR2 in hippocampal neurons can activate the autophagy pathway by up-regulating JNK signal pathway, thus attenuate oxidative stress injury and protect neurons.]]> 2021/7/28 0:00:00 YANG Yang,ZHANG Chen,FENG Lu-Qiu,XIE Qing,YANG Cheng-Ying and GAN Ling 245true 2021/7/28 0:00:00 HU Yi-Fan,FENG Zhou-Yan,WANG Zhao-Xiang and ZHENG Lü-Piao 244true 2021/7/28 0:00:00 WANG Yu-Xiang,JIANG Shuang,SHI Ya-Ning,ZHANG Chan-Juan,LIU Le-Ping,ZHAO Tan-Jun,GONG Yong-Zhen,LIAO Duan-Fang and QIN Li 243true Tm value of primer sequences, how to remove false positive amplification and how to improve sensitivity. Though most MSP primer design tools have proposed various solutions for those difficulties, there are still some defects in consideration of primer influencing factors, multitasking, prediction of specific amplification in MSP primer design and evaluation. Therefore, in this study, after deep exploration of existing MSP primer design tools, a novel MSP primer design and graphic evaluation tool named MethyScan was developed based on the integration of Bowtie, SAMtools, and BEDTools with Python graphic library Matplotlib and third-party functional libraries Biopython and Primer3-py. Three functional modules were involved in MethyScan including primer design, genome indexing and primer evaluation. MethyScan not only has the ability to perform MSP primers design and Nested primers adaptation, but also can evaluate primers specific/non-specific amplification with the analysis of primer binding information on four converted genomic templates and graphically displaying of the difference between non-specific amplification and target. Meanwhile, the comparison of MSP primer design for six potential biomarkers TFPI-2, NDRG4, CDKN2A, CD44, CASP8, and SDHD in esophageal cancer, colorectal cancer, and other malignant tumors suggested that MethyScan can not only obtain primers with more CpG sites, but also obtain primers with same or similar locations to those of other softwares, and the difference of Tm values of primers is even smaller. As the first MSP primer design tool for graphically displaying specific/non-specific amplification differences, MethyScan can effectively improve the accuracy of methylation primer design and provides strong support for the development of clinical DNA methylation detection projects, tests and diagnostic kits. The download address of MethyScan is: https://github.com/bioinfo-ibms-pumc/MethyScan.]]> 2021/6/24 0:00:00 CAO Ying-Hao 242true Xanthomonas campestris pv. campestris (Xcc) causes black rot disease to all cruciferous plants, and great economic losses to the world. To study the 3-hydroxylacyl-ACP dehydratase in Xcc, XC_2876 (XcfabZ) was found based on the sequence alignment with E. coli FabZ. Moreover, XcFabZ shows 46.1% amino acid sequence identity with EcFabZ, and contains the conservative α-helix structure and active residue. XcfabZ was able to genetically complement the EcfabZ knock out mutant E.coli HW7 to restore the growth with inducer IPTG. In vitro assay also identified XcFabZ was able to dehydrated 3-hydroxyacyl-ACP in the initial and elongation reactions. However, XcfabZ in the chromosome could not be deleted directly, indicating XcfabZ is essential for growth. When expression plasmids harboring EcfabZ or XcfabZ was introduced, respectively, XcfabZ deletion mutants were constructed. The EcfabZ replaced mutant showed different fatty acid compositions, much lower tolerance to stressful conditions (high salty, low pH, H2O2 and SDS), and decreased motility compared to the wild-type. While mutant with XcfabZ in the plasmid showed similar phenotype to wild-type. These data demonstrated both XcFabZ and EcFabZ show 3-hydroxylacyl-ACP dehydratase activity, but they may contains different biological function in vivo.]]> 2021/6/24 0:00:00 MA Jian-Rong,CHEN Cheng,YAN Ming-Feng,LI Xian-Qi,ZHANG Wen-Bin and YU Yong-Hong 241true 2+]i amplitude, [Ca2+]i fluorescence ratio, departure velocity, time to peak(TTP), time to peak50%(TTP50%), time to baseline50%(TTB50%), return velocity, ratio amplitude, maximum contraction and diastolic rate of sarcomere(±dl/dtmax), sarcomere length(SL), peak twitch amplitude(PTA) and SL shortening%. The related proteins of PI3K-AKT-PKG-1/p-PLN-SERCA2a signaling pathway and apoptosis were detected by PI3K-AKT-PKG-1/p-PLN-SERCA2a signaling pathway and apoptosis related proteins were detected by Western blotting. Compared with group S, in group MI, PI3K-Akt-PKG-1/p-PLN-SERCA2a signaling pathway is significantly inhibited, apoptosis and left ventricular end-diastolic pressure (LVEDP) are significantly increased, left ventricular systolic pressure (LVSP) and maximum pressure increasing and decreasing rate(±dp/dtmax), [Ca2+]i amplitude, [Ca2+]i ratio amplitude, departure velocity and return velocity all significantly reduce, and TTB50%, TTP and TTP50% both significantly increase. The myocardial cells were significantly reduced at the SL shortening%, PTA, ±dp/dtmax. Compared with MI group, PI3K-Akt-PKG-1/p-PLN-SERCA2a signaling pathway in ME group is significantly activated, apoptosis and LVEDP are significantly reduced, LVSP and ±dp/dtmax are significantly increased, ratio amplitude, [Ca2+]i amplitude, ratio velocity and departure velocity are significantly increased, and TB50%, TTP and TTP50% are significantly shortened. The myocardial cells were significantly inaeased at the SL shortening%, PTA, ±dp/dtmax. It can be concluded that aerobic exercise improves the calcium transient and myocardial systolic function in peripheral infarction area of MI rats, activates the PI3K-Akt-PKG-1/p-PLN-SERCA2a signal, inhibits the apoptosis of myocardial cells, and improves myocardial infarction function. Moreover, the improvement of myocardial infarction function is closely related to the activation of PI3K-Akt-PKG-1/p-PLN-SERCA2a signaling pathway and the inhibition of myocardial cell apoptosis.]]> 2021/6/24 0:00:00 LIU Niu and TIAN Zhen-Jun 240true rd/rd, cl/cl mice as visual weakening group. Freezing behaviors were recorded during fear conditioning, fear extinction, and extinction recall phases. The results indicated that mice with shape vision deprivation are more conducive to auditory fear conditioning. We discuss the possible neuro-mechanism of visual-auditory system interactions.]]> 2021/6/24 0:00:00 FU Juan,ZHAO Xu-Dong,LIU Tian-E,MA Man-Xiu,SUN Hua-Ying,ZHANG Bo and ZHENG Xi-Geng 239true 2021/5/31 0:00:00 Zhou Zi-hua,Yue Chun-xue,Liang Lin,Gao Lu,He Zheng-xi,He Jun-yu,Jin Xi,Gao Meng-xiang,Li Xin and Zhou Yan-hong 238true 2021/5/31 0:00:00 LIU Jin-Wei,CAO Gao-Biao,CHENG He-Yun,DU Wei-Wei,ZHANG Guang-Liang,JIN Guang-Zhe,ZHANG Ping,WANG Zhao-Dong,LIU Zhe,JU Ji-Hui,LE Ying-Ying,FU Yi and HOU Rui-Xing 237true Streptomyces, they have been used to synthesize many biologically active secondary metabolites, such as antibiotics, anti-tumor drugs and immunosuppressants. Due to the fact that the synthesis of these products is often strictly regulated at multiple levels, therefore, the study of the mechanism of secondary metabolism regulation of Streptomyces can not only deepen our understanding of the metabolic regulation network of Streptomyces, but also provide important reference and guidance for the construction of industrial producing strains from the perspective of metabolism. There are two key types of signal transduction system in Streptomyces: two-component system (TCS) and extracytoplasmic function σ (ECF-σ). Both of them play important regulatory functions in the process of antibiotic biosynthesis. Studies have shown that there are a large number of TCS and ECF-σ coding genes in the genome of Streptomyces coelicolor, a model Streptomyces strain. In our previous studies, we have showed that, under certain conditions, the sigQ-afsQs gene cluster in S. coelicolor is involved in the regulation of the biosynthesis of ACT (actinorhodin), RED (undecylprodigiosin) and CDA (calcium-dependent antibiotic) antibiotics. Based on the early stage research on the regulation function of the TCS system afsQ1/Q2, a detailed study of the upstream regulation mechanism toward afsQ1/Q2 was carried out in this work. Through gene function verification experiments, it was found that the loss of sigQ can significantly down-regulate the expression of the membrane protein gene afsQ4 in the sigQ-afsQs gene cluster, and at the same time complementation afsQ4 can restore the phenotype of the sigQ deletion mutant (?sigQ), which indicates that afsQ4 is the downstream regulatory target of sigQ. Further analysis of in vitro phosphorylation experiments showed that the phosphorylation level of the transmembrane kinase AfsQ2 of TCS was significantly reduced in afsQ4 gene deletion mutant, indicating that sigQ can negatively regulate the TCS afsQ1/Q2 through membrane protein AfsQ4, and finally coordinate antibiotic synthesis.]]> 2021/5/12 0:00:00 CHEN Yun-Liang,YANG Yun-Peng,LI Guo-Quan,MAO Xue-Fang,JIA Wei-Dong,SHI Ai-Ping and LU Yin-Hua 236true 2021/5/12 0:00:00 CAI Yang,LI Bao-Wei and ZHOU Yi-Feng 235true in vitro was also retarded. Knockdown of EF4 down-regulated the expression of mitochondrial DNA-encoded subunits of electron transfer chain complexes, and resulted in the dysfunction of mitochondrial oxidative phosphorylation. These results define a tumor-supportive role for EF4 by maintaining the protein synthesis within the mitochondria, which may serve as a potential therapeutic target in bladder urothelial carcinoma.]]> 2021/4/26 0:00:00 HUA Chao-Ju,DAI Fei,DENG Yao,LU Jun-Wan,ZHOU Ning-Ning,LI Wan-Qi,ZHANG Ye,HUANG Ka-Te,LIU Yong-Zhang,Lü Bin and WEI Tao-Tao 234true 2021/4/26 0:00:00 SUN Kai,WEI Qing-Gong,ZANG Chao-Yu,SUN Ru-Xuan,JIANG Dan and SUN Xiao-Yong 233true Ash2l promotes H3K4 trimethylation levels through methyltransferase MLL/SET1 complex, which is essential for mouse embryonic development. Ash2l have two isoforms (Ash2l-1 and Ash2l-2) through alternative promoter usage in mouse. However, the mechanism of this gene in mouse embryonic development and the function of different isoforms remain unknown. In this study, we used CRISPR/Cas9 technology to specifically disrupt Ash2l-1 in mice and investigated the role of Ash2l-1 in mouse early embryonic development. Disruption of Ash2l-1 resulted in embryonic lethality at E9.5-10.5. Particularly, E9.5 Ash2l-1-deficient embryos exhibited severe growth defects, including developmental defects of yolk sac. Gene expression profiling showed that Ash2l-1 deficiency affected the expression of specific genes involved in erythropoiesis and vascular formation. CUT&RUN analysis showed that H3K4me3 levels of the promoter of some specific genes involved in erythropoiesis and vascular formation was down-regulated. Taken together, these results indicate that Ash2l-1 regulates gene expression through H3K4me3 and plays an essential role in the early hematopoiesis of mouse yolk sacs.]]> 2021/4/8 0:00:00 WANG Xiao-Qing,LU Xu-Kun,XIANG Yun-Long and LI Lei 232true In situ self-assembly peptides which can target the tumors and self-assemble into specific nanostructures induced by the specific and highly expressed substances at the tumor tissues are expected to be a new kind of anti-tumor drugs. In this study, we designed a peptide denoted as Fmoc-FFRIKFERQ-OH based on the concept of in situ self-assembly of peptides. This peptide can target lysosomes and self-assemble in situ after being degraded by cathepsin L in lysosomes. The self-assembly properties of Fmoc-FFRIKFERQ-OH and its enzymolysis products degraded by cathepsin L were oberved by AFM and MALDI-TOF-MS. The anti-tumor activity of this peptide was measured by using the MTT and flow cytometry assay. The results demonstrated that cathepsin L could precisely hydrolyze the Fmoc-FFRIKFERQ-OH molecule under acidic conditions in vitro, and its enzymolysis product Fmoc-FFR-OH self-assembled into long nanofibers, which had high antitumor activity against both A375 and SH-SY5Y cells. The relative survival rate was only 36.08% and 25.56% for A375 and SH-SY5Y, respectively. Comparatively, Fmoc-FFRIKFERQ-OH had lower toxic and side effects on normal cells L929. The self-assembled long fibers formed by Fmoc-FFR-OH can disrupt the membrane of lysosome, which can induce cell apoptosis and(/or) death. Thus, the designed Fmoc-FFRIKFERQ-OH is expected to be a new anti-tumor drug due to its high antitumor activity and low toxicity against to normal cells.]]> 2021/4/8 0:00:00 CHEN Cui-Xia,PENG Xiao-Ting,ZHU Yu-Ting,HAO Rui-Rui,YANG Liu-Xin,WANG Tong,NIU Xiao-Ya and ZHAO Yu-Rong 231true Moraxella bovoculi is an inhibitor of Type V-A CRISPR-Cas system. However, the structure and inhibition mechanism of AcrVA2 remain to be elucidated. Here we report the crystal structures of AcrVA2 in the apo state and MbCas12a620-636-AcrVA2 complex. AcrVA2 adopts a novel α-β fold and binds to MbCas12a in free state. The structure of MbCas12a620-636-AcrVA2 complex reveals that AcrVA2 interacts with MbCas12a via hydrogen bonds and salt bridges, as well as hydrophobic interaction. These results suggest that AcrVA2 affect the activity of Cas12a by binding to the MbCas12a in the apo state. These results provide significant insights into the mechanism of AcrVA2 disabling Type V-A CRISPR-Cas system.]]> 2021/4/2 0:00:00 CHEN Peng,SUN Wei,CHENG Zhi,YANG Jing,WANG Min,WANG Jiu-Yu,CHEN Hui-Qing,LIU Liang and WANG Yan-Li 230true Lactobacillus acidophilus is a kind of probiotics. The previous research found that β-glucosidase (BGL) produced by Lactobacillus acidophilus GIM1.208 has high activity. In order to elucidate its structure and characteristics, the target gene of Lactobacillus acidophilus GIM1.208 BGL was obtained with PCR and successfully expressed in E. coli. Protein samples with a purity of more than 90% was obtained by nickel affinity chromatography. Then the secondary structure of BGL was detected by circular dichroism spectroscopy (CD) and its tertiary structure was analyzed by homology modeling method. The enzymatic properties of Lactobacillus acidophilus BGL was also studied. The results show that the molecular mass of Lactobacillus acidophilus BGL is 52 ku, the concentration after purification is 1.88 g/L. The secondary structure of Lactobacillus acidophilus BGL includes 15.9% α-helix, 44.1% β-sheet, 18.1% β-turn, and 27.2% random coil. Homology modeling analyzation showed that 8 β-sheets and 8 α-helices were included in 3D structure of Lactobacillus acidophilus BGL, and the whole protein is conical in shape. Lactobacillus acidophilus BGL has good glucose tolerance and NaCl tolerance. The optimal temperature and the optimal pH of this enzyme is 47℃ and 5.6, respectively. Lactobacillus acidophilus BGL has high stability in the range of 20℃-50℃ and pH 2.2-6.0. The inhibitory effect of ethyl acetate and methanol on enzyme activity is obvious, and Fe3+ and Fe2+ have significant activation effect on enzyme. These results provide important reference basis for the subsequent functional exploration and application research of Lactobacillus acidophilus BGL.]]> 2021/4/2 0:00:00 YU Yi-Hong,DING Xiao-Juan,DING Zhu-Hong,SONG Yu-Ting and WANG Yi 229true 2021/12/23 13:40:56 DENG Xue-Qing,WANG Shi-Hao,GONG Wei-Kang and LI Chun-Hua 228true Lon/Swe transgenic mice, which showed memory deficiency at 3 months of age but normal memory at 6 months. Impaired memory in 3-month-old mice was accompanied by higher levels of formaldehyde and hyperphosphorylated tau than controls. Administration of resveratrol, which is a formaldehyde scavenger, rescued the cognitive deficits in these mice by reducing formaldehyde levels and attenuating tau hyperphosphorylation. With increased expression of formaldehyde catalytic enzymes such as aldehyde dehydrogenase 2 (ALDH2) and alcohol dehydrogenase III (ADH3), 6-month-old AβPPLon/Swe mice displayed similar levels of formaldehyde and working memory as controls. We discovered that brain formaldehyde levels were significantly associated with the progression of memory deficit in AβPPLon/Swe transgenic mice, and that recovery of memory was associated with formaldehyde reduction. Our findings provide valuable insights into the underlying mechanisms of AD.]]> 2021/11/23 0:00:00 LU Jing,HE Huan,MIAO Jun-Ye,ZHU Yan,LI Ting,CHEN Xi-Xi,TONG Zhi-Qian,HE Rong-Qiao and LIU Ying 227true 2021/11/23 0:00:00 YAO Ze-Huan,CHEN Wei,LI Chen,YANG Hao-Yi,HE Yu-Lin,TAN Yu-Song and LI Fei 226true via caveolae-mediated pathway rather than clathrin mediated pathways or macropinocytosis. Our study provided a potential probe for nucleolus and this finding is of great significance for the development of functional nanomaterials targeting subcellular organelles.]]> 2021/11/26 0:00:00 WANG Qian,FENG Zhen-Zhen,MAO Jian,LIU Li-Hua,CHEN Xiao-Liang,WEI Xiao-Yun and HUANG Fang 225true M. jannaschii tyrosyl-tRNA synthetase (Mj. TyrRS) mutants reported in the literature to compare and analyze the sequence and structural feature and difference between mutant and wild type Mj. TyrRS. Mj. TyrRS is used as an example since the sequences and structures of many unnatural amino acid specific Mj. TyrRS mutants have been reported. Based on the crystal structures of different Mj. TyrRS mutants and Rosetta modeling result, we found D158G/P is the critical residue which influences the backbone disruption of helix with residue 158-163. Our results showed that compared with random mutation, Rosetta modeling and score function calculation can elevate the enrichment ratio of desired mutants by 2-fold in a test library having 687 mutants, while after calibration by machine learning model trained using known data of Mj. TyrRS mutants and ligand, the enrichment ratio can be elevated by 11-fold. This molecular modeling and machine learning-integrated workflow is anticipated to significantly benefit to the Mj. tyrRS mutant screening and substantially reduce the time and cost of wet-lab experiments. Besides, this novel process will have broad application in the field of computational protein design.]]> 2021/11/26 0:00:00 DUAN Bing-Ya and SUN Ying-Fei 224true 2021/11/26 0:00:00 WANG Xiao-Xi,LI Xiao-Qin,CAO A-Cheng,HOU Zhi-Chao and GAO Bin 223true Objective To investigate the effect and mechanism of Zuogui pill-containing serum (ZGP-containing serum) on chemotherapy-damaged granular cells (GCs) and theca cells (TCs).Methods GCs and TCs were cultured respectively, the model group was established with phosphoramide mustard (PM), then treated by ZGP-containing serum. The survival rates of GCs and TCs was determined by CCK-8. Real-time fluorescent quantitative PCR (RT-PCR) and Western blot methods were used to detect the expression of Beclin-1, light chain 3 (LC3B), p62, Bax and Caspase3.Results 10% ZGP-containing serum had the best effect on the recovery of cell survival rate. Compared with the blank control group, Beclin-1, LC3B, Bax and Caspase3 have higher expression in the model group (P<0.05), and 10% ZGP-containing serum can down-regulate the expression of them (P<0.05). Moreover, the expression of p62 is lower in the model group than the blank control group (P<0.05), and 10% ZGP-containing serum can up-regulate it (P<0.05). In addition, in the groups of GCs, after activating or inhibiting the autophagy pathway, the expression of autophagy-related proteins and apoptosis-related proteins both changed correspondingly.Conclusion PM can damage GCs and TCs by promoting apoptosis and activating autophagy/lysosomal degradation pathways, 10% ZGP-containing serum can alleviate the damage. There is a cross-talk between autophagy and apoptosis in the process of PM damaging GCs which can be alleviated by 10% ZGP containing serum.]]> 2022/9/21 0:00:00 LIU Tao,SUN Xiao-Feng,ZENG Gui-Rong,HUANG Xin-Yi,ZENG Wen-Zhuo,GONG Li-Min,YANG Song-Wei and LIU Meng-Lin 222true Objective Dendrobium officinale has long been used as an important medicinal herb in oriental medicine. Polysaccharide, flavonoid, and alkaloid are the major active ingredients, the production and accumulation of which are frequently affected by numerous environmental cues. Phytohormone supplemented in culture medium has facilitated the mass production of orchids. However, their mechanism of action on the production of active components in Dendrobium officinale is far from clear.Methods Here, major medicinal metabolites were comparatively analyzed in Dendrobium officinale seedlings exposed to the most commonly used phytohormones (NAA and/or 6-BA), and transcriptomes corresponding to the treatments were generated.Results Results showed that phytohormones greatly affected the accumulation of polysaccharide, alkaloid, and flavonoid, and triggered tremendous transcriptional changes. It was demonstrated that 6-BA induced more transcripts than NAA and that β-glucosidase (BGLU) expression was closely related to polysaccharide production. Further functional analysis revealed that factors including phytohormone category, concentration, treating duration, and seedling growth stages, can drastically affect the BGLU expression and the corresponding polysaccharide production, thus partially answered the key question at molecular level why the medicinal constituents are unstable in tissue culture derived Dendrobium plants.Conclusion Altogether, the present study clearly demonstrated that BGLU is a key regulator for polysaccharide production in Dendrobium officinale in response to phytohormone treatments.]]> 2022/9/21 0:00:00 WANG Zhi-Cai,ZHAO Mei-Li,ZHANG Xiao-Jie,ZHANG Zhen-Liang,LI Su-Zhen,LI Jian,CUI Hong-Qiu,LI Wu-Jiao,LIU Yan-Chun,WANG Yu,LI Li-Qiang,GU Li-Li and WANG Mei-Na 221true Objective TP53 is an important tumor suppressor gene, however, mutations of p53 occur in over 50% of all cancers and are indicative of highly aggressive cancers that are hard to treat. Targeting mutant p53 (mutp53) is one of the effective strategies in cancer therapy.Methods We constructed a high throughput screen system which contains the p53 targeted puma, H1299-p53R273H-PUMA-luciferase, and H1299-p53R175H-PUMA-luciferase to screen compounds targeting mutp53. Using immunofluorescence assay to detect the effect of andrographolide on the expression of mutp53 in HT29 (R273H) and SK-BR-3 (R175H) cells. Western blotting experiment and qRT-PCR analysis were used to further observe the protein and mRNA expressions of andrographolide on p53 downstream target genes PUMA, p21, and Noxa in mutp53 tumor cells. Then MTT and flow cytometry were used to detect andrographis paniculata The effect of lactone on tumor cell proliferation and apoptosis. In addition, after knocking down Hsp70 expression by siRNA, the reactivation effect of andrographolide on mutp53 downstream genes was also studied.Results Andrographolide enhanced PUMA-luciferase expressions in both cell lines. Andrographolide could reduce the expressions of mutp53 in HT29 (R273H) and SK-BR-3 (R175H) cells, while the expressions of wild-type p53 increased by immunofluorescence assay. Andrographolide can inhibit the cancer cell proliferation and induce apotposis in mutant p53 cancer cells. Andrographolide can enhance the p53 downstream target protein and mRNA expressions of PUMA, p21, and Noxa. Andrographolide increased the expression of molecular chaperone Hsp70 in HT29 and SK-BR-3 cells, as the chaperones play important role in p53 structure function, we knocked down Hsp70 by siRNA and found that the upregulation of p53 targeted genes was reversed.Conclusion Andrographolide restores the wild-type-like p53 function dependent on Hsp70.]]> 2022/9/21 0:00:00 SONG Bin,WANG Jia-Jian,SU Yong-Nan,WANG Yu-Ling,YANG Fan and ZHANG Ji-Hong 220true Objective Hair shaft is one kind of common biological evidences at the crime scene. However, it fails to play an important role in the crime investigation due to lack of effective method of individual identification. The single amino acid polymorphisms (SAPs) in the hair shaft proteome contain information of individual genetic differences.Methods In order to study SAPs in the hair shaft, the proteome of single 2 cm hair shaft samples (6 people, 2 hairs per person) were extracted using ionic liquid following with LC-MS/MS detecting. The protein composition of the hair shaft was analyzed. A custom SAP protein sequence database was built for East Asian population as the searching database. Based on the custom SAP and SNP corresponding annotation table information, the nsSNP profiles were imputed corresponding to SAP. The accuracy of SAP was verified by comparing the imputed nsSNP profiles from SAP with nsSNP profiles obtained from the whole exome sequencing. The validated SAPs were used to calculate the random matching probability.Results In 12 samples, 321 SAPs were obtained, with an average of (131±17) for each sample. The value of random matching probability for 6 people ranged from 1.4×10-4 to 1.0×10-9.Conclusion In this paper, a method for detecting SAP in hair shaft proteins of East Asian populations was established, and the ability of individual identification application was verified, which provided a powerful tool and new ideas for individual identification of hair shaft in forensic science.]]> 2022/9/21 0:00:00 WU Jia-Lei,JI An-Quan,DING Dong-Sheng,FENG Lei and YE Jian 219true Objective As for the unique properties of good biocompatibility and excellent optical performance, silica nanotubes have shown potential applications in different fields. The properties of the silica nanotubes were well correlated with their size and morphologies. In order to prepare silica nanotubes with much larger size and expand their applications in different fields, this study selected wide peptide nanotubes as templates for biomimetic synthesis of silica materials.Methods In order to obtain silica nanotubes with relative large size, here we prepared silica nanotubes by using wide nanotubes (with diameters of approximately 40 nm) self-assembled by the bola peptide Ac-KI3VK-CONH2 as templates through a biomimetic mineralization route. The stability of the peptide nanotubes was firstly investigated and the results demonstrated that these nanotubes can be easily destroyed by dilution, addition of organic solvents, or changing the solution pH value, showing poor stability. Then, we chose methyl n-silicate (TMOS) with fast reaction rate as the precursor to prepare silica nanomaterials and systemically explored the influence of different factors on the size and morphology of the silica nanotubes.Results The results demonstrated that the ideal condition for preparing silica nanotubes with uniform shape and size was using TMOS as precursor and keeping its concentration at 1.11%-3.33% (v/v) under neutral or weak alkaline conditions.Conclusion This work successfully prepared silica nanotubes with relative large diameter by using self-assemblies formed by a bola peptide as templates and silicon source with fast reaction rate, which is of great significance and may expand the applications of wide silica nanotubes in different fields.]]> 2022/9/21 0:00:00 ZHAO Yu-Rong,WANG Li,LI Xing-Fan,ZHANG Li-Min,WANG Dong and CHEN Cui-Xia 218true Objective Based on chemical structural characteristics, the bioactivity of polyoxypregnane glycosides from Dregea sinensis, and the molecular interaction between compounds and targeted proteins were investigated.Methods The bioactive screening of 191 polyoxypregnane glycosides (>800 u) , and the kinetic evaluation on human immune-related proteins was carried out by molecular docking and SPR experiment.Results Seven compounds (6, 18, 23, 30, 78, 79, and 80) and 3 immune-related proteins (IL-2Rα, TLR4, and TNF-α) were selected through virtual screening. Compounds 30 and 78 showed the significant binding tendency with IL-2Rα and TLR4 in SPR experiment, and KD values with IL-2Rα were 2.41×10-6 and 2.14×10-6 mol/L, meanwhile KD values with TLR4 were 1.96×10-5 and 5.60×10-6 mol/L, respectively. The interactions between targeted proteins and compounds were further characterized by discovery studio. The analysis of molecular docking revealed the binding pocket residues for SPR-positive molecules 6, 18, 30, 78, and 80.Conclusion The result indicated these glycosides could bind to targeted proteins through forming hydrogen bonds and Pi-Pi interactions. The study is meaningful for bioactive evaluation of polyoxypregnane glycosides, and provides valuable exploration to the underlying mechanisms of effective compounds with low abundance.]]> 2022/9/21 0:00:00 WANG Zhi,SONG Juan,DAI Rong-Ji,DENG Yu-Lin and Lü Fang 217true Objective Chemotaxis and electrotaxis are the primary mechanisms underlying directed cell migration, and they play important roles in the physiology and pathology of organisms. However, there are differences between the two. This paper presents a comparative study of the roles of Dictyostelium discoideum genes gbpC and gbpD in cell electrotaxis and chemotaxis in order to gain further insight into the differences between these two mechanisms of migration.Methods The gbpC/gefT- mutant strain and gbpD/gefU- mutant strain were placed in a 12 V/cm direct current (DC) electric field to investigate the direction and velocity of cell movement and the changes in cell electrotaxis; Lifeact-GFP (F-actin) was electroporated into cells and the distribution of F-actin during cell movement was observed under a fluorescence microscope; Western blot was used to quantify the phosphorylation of myosin regulatory light chain (RLC) in cells stimulated by DC electric fields.Results The gefT- mutant cells lost polarization but retained electrotaxis at a level similar to the wild-type cells; the gefU- mutant cells showed hyperpolarization but significantly reduced electrotaxis; in a DC field, F-actin was predominantly distributed in the pseudopods in both mutant and wild-type cells; there were differences in myosin RLC phosphorylation between the cell lines in electric fields. Phosphorylation was time-dependent in wild-type cells, whereas phosphorylation first decreased rapidly and then increased in gefT- mutant cells. Time-dependent dephosphorylation occurred in gefU- mutant cells.Conclusion Our findings indicate that gbpC and gbpD play differntal roles between the chemotaxis and electrotaxis of Dictyostelium discoideum and provide further evidence that electrical and chemical signals drive the directed migration of cells through different mechanisms.]]> 2022/8/19 0:00:00 JIANG Rui-Da,WANG Jia-Jia,Zhao San-Jun,WANG Xiao-Yan and GAO Run-Chi 216true Objective The present study was to clarify the improving effect of artemisinin on diabetes-induced cognitive deficit and the underlying mechanisms in mice.Methods Type 2 diabetes mellitus (T2DM) mouse models were established by a single dose of STZ injection (100 mg/kg, i.p.). Those animals were then treated with vehicle or artemisinin (40 mg/kg, i.p.) once daily for 4 weeks. Cognitive performances of the mice were evaluated by novel object recognition, Y maze test and Morris water maze test. After behavioral tests, the expressions of PI3K, Akt, SYN and PSD-95 proteins in the hippocampus were measured by Western blot. Changes in the synaptic ultrastructure of the hippocampal CA1 region were observed by transmission electron microscope.Results Our results indicated that artemisinin significantly ameliorated cognitive deficit in T2DM mice. Furthermore, PI3K and phosphorylated Akt protein levels in the hippocampus of T2DM mice treated with artemisinin were elevated, accompanied with increases in the number of hippocampal neurons, as well as the protein contents of SYN and PSD-95. Meanwhile, synaptic plasticity was also rescued, indicated by an increase in synapse number and synaptic curvature, the thickness of postsynaptic density, and a decrease in the width of synaptic cleft in the hippocampal CA1 region.Conclusion Taken together, these results demonstrate that artemisinin can protect T2DM mice against cognitive decline, at least partially through activating PI3K/Akt pathway to improve synaptic plasticity in the hippocampus. These findings demonstrate that artemisinin may serve as a novel therapeutic agent for diabetic cognitive impairment.]]> 2022/8/19 0:00:00 QIU Ming-Yue,LUO Yan,LI Shao-He,NIE Ya-Xiong,CHEN Ru-Meng,TANG Ya-Ling,LI Chao and GU Hong-Feng 215true Objective More than 200 million people are visually impaired or blind worldwide. Artificial vision system has been widely studied for a long time. There are two main technical paths for the research of artificial vision system, the first is implantable artificial vision device, and the second is non-implantable and wearable device. This study demonstrates a non-implantable system prototype based on a wearable touch-vision device designed for the head that can help blind people and visually impaired people complete complex tasks in life such as walking.Methods The image information front of the subjects is collected, and transmitted wirelessly to the operator. After analyzed and processed, it is wirelessly transmitted to wearable devices such as headgears in the form of tactile coding to trigger the multi-point head tactile sensation, thus the subject’s information or precise action instructions is obtained. The system also assists the subjects with voice information, allowing them to learn more about road conditions and environmental information.Results 5 healthy volunteers (2 males and 3 females) and 1 young blind person (male) were tested with the prototype. The results show that, the prototype makes full use of the head’s natural distinguishing ability for directions including front, back, left and right, and quick response ability to the touch, and the prototype can send clear instructions such as going straight, adjusting to the left or right, and turning. The test showed that the response time of subjects to tactile commands was within 0.5 s, and the mean value of the standard deviation of the deviation of walking for about 7 m is reduced to (16± 10) cm compared with the case without wearing the device.Conclusion Experimental results show that the system can provide environmental image information in a concealed and accurate manner, and help visually impaired people complete daily activities such as walking, avoiding obstacles, going up steps, entering cafes and other public places, fetching objects on the table, which meets expectations of assisting blind people improve their quality of life. In the future, the prototype system will be further improved, especially miniaturization and smart chip. The tactile devices are made into neck rings, belts, bracelets, foot rings, etc., which will be widely used in a variety of work scenarios such as travel assistance for the blind, night field trips, and deep sea diving.]]> 2022/8/19 0:00:00 GE Song,LIN Yan-Ni,LAI Shun-Nan,XU Jing-Jing,HE Yu-Li,ZHAO Qi,ZHANG Hong and XU Sheng-Yong 214true Objective To investigate the effects of lactoferrin (LF) deficiency during lactation and LF supplementation from different sources on intestinal development of newborn mice.Methods Female LF knockout mice were used to produce LF deficiency milk for newborn mice intake during lactation, and 100 mg/kg bovine Lactoferrin (bLF), recombinant human Lactoferrin (rhLF) and bovine serum albumin (BSA) were artificially fed every day from the 3rd day to the 21st day. Samples were taken at the age of 21 days to determine the small intestine development index of mice in each group.Results In this experimental cycle, rhLF supplementation significantly increased the villus length/crypt depth of ileum (P<0.05), up-regulated the expression of Occludin and ZO-1 in ileum (P<0.05) and increased the ratio of maltase activity/lactase activity in duodenum (P<0.05), jejunum (P<0.05) and ileum (P<0.05) of mice. In addition, the supplementation of bLF only significantly increased the ratio of maltase activity/lactase activity in ileum (P<0.05), indicating that the supplementation of LF during lactation can promote the intestinal development of newborn mice.Conclusion For newborn mouse without LF intake during lactation, the supplement of LF during lactation can enhance the intestinal digestion and absorption of nutrients, promote the development and maturity of the intestinal tract, and enhance the intestinal barrier function. Moreover, rhLF plays a more effective role than bLF.]]> 2022/8/19 0:00:00 CHENG Zhi-Mei,WANG Wen-Li,LI Ze-Yang,MENG Qing-Yong,DAI Yun-Ping and ZHANG Ya-Li 213true Objective Quick TarSeq integrated system developed by our groups for on-field fast analysis is a fully automated and integrated system, which can be applied to the InDel ancestry inference, and complete the “sample-in result-out” rapid automatic InDel typing in about 2 h. In this paper, the performance of the InDel ancestry inference microfluidic chip amplification system was studied, hoping to provide a reference for practical application.Methods The system was validated by sensitivity, inhibitor tests, success rate, concordance tests, sizing precision and accuracy, peak height balance, case sample tests, and the accuracy of infering ancestral origins.Results The microfluidic chip amplification system was concordant with traditional methods, with 95.65% success rates and 98.85% concordance rates for 138 testing samples. The complete InDel profiles can be obtained from more than 5 ng of template DNA input. The best collection of buccal swab was the donor who performed 8 swipes inside of cheek. The best collection of dried blood spot samples was 6 pieces (Φ=2 mm). The average heterozygous peak height ratio across all locus was 0.86. The standard deviation of allelic ladder for 10 runs was within 0.3 bp. The size differences between allele and corresponding allele in allelic ladder was within 0.5 bp.Conclusion Microfluidic chip amplification system enable providing reliable InDel profiles from buccal swabs, dried blood spot, dried saliva spot samples and cigarette butt with accurately ancestral inference. It is possible to yield “sample-in result-out” results of InDel typing in 2 h. This system can meet the requirements of on-site testing and can be used for forensic casework.]]> 2022/8/19 0:00:00 ZHANG Bo-Yuan,WANG Ying-Xi,ZHAO Lei,JIANG Li,WAN Qun,ZHUANG Bin,ZHAO Li-Jian,YANG Rui-Qin and HAN Jun-Ping 212true Objective As a new type of transcranial magnetic stimulation, the intermittent theta burst stimulation (iTBS) has been widely used to explore brain cognitive function and neuroregulation, but its electrophysiological regulation mechanism is still unclear. Therefore, exploring the underlying electrophysiological mechanism and the effect of iTBS on the brain’s cognitive functions is of great significance for the treatment of brain diseases and clinical application of magnetic stimulation.Methods In this study, the model rats were treated with iTBS, and the local field potential (LFPs) signals in the ventral hippocampus (vHPC) and medial prefrontal lobe (mPFC) were collected and recorded during the working memory (WM) tasks in rats. The effect of iTBS on neural network coordination and information interaction across brain regions was investigated by using Granger causality network.Results The results suggested that iTBS enhanced the WM ability and the behavioral performance of the rats, reducing the time to complete the WM task by (2.67±1.63) d (P<0.05). At the same time, iTBS enhanced the self-causal network connections in vHPC and mPFC during WM, and increased the network connection strength, connection density and global efficiency (P<0.05). In addition, iTBS enhanced the cross-brain network connection between vHPC and mPFC, and increased the node degrees and causal flow (P<0.05) from vHPC to mPFC.Conclusion iTBS has a significant positive effect on the behavior of WM and neural network in related brain areas of rats. iTBS can promote the cognitive ability of rats and improve the efficiency of information interaction and transmission of neural network. The neural regulation mechanism of iTBS may be to enhance the WM ability by enhancing the network connection and information interaction between vHPC and mPFC.]]> 2022/8/19 0:00:00 ZHANG Tian-Heng,GUO Miao-Miao,XU Gui-Zhi,JI Li-Hui and WANG Zhong-Hao 211true Objective PAMM (peroxiredoxin-like 2 activated in M-CSF stimulated monocytes) is a secreted protein which shows high expression in white adipose tissues, but the roles of PAMM in many biological processes are still unknown. To provide new clues for PAMM function research, this study is intended to investigate the possible role of PAMM in white adipogenesis as well as the downstream genes regulated by PAMM.Methods Adipogenic differentiation and adipogenic inhibition models of human ADSCs (adipose-derived stem cells) were established using adipogenic cocktail (AC) medium or AC plus IL-1α, respectively. Expression of PAMM in ADSCs was suppressed or overexpressed using siRNA interference or plasmid transfection. Gene array, mRNA sequencing and quantitative RT-PCR were employed to detect the mRNA expression level. Western blot was used to evaluate protein expression and Oil red O staining was adopted to assess lipid droplets accumulation.Results Expression of PAMM was increased following white adipogenic differentiation of ADSCs and decreased following adipogenic inhibition. However, when PAMM was knocked down or overexpressed before adipogenic differentiation of ADSCs, the downregulation and upregulation of PAMM expression generally did not exert evident influence on the formation of lipid droplets and the expression of adipogenesis-related genes. Similarly, PAMM knockdown in highly differentiated adipocytes had no obvious effect on cellular morphology and the accumulation of lipid droplets. Finally, a bunch of functional genes and gene sets regulated by PAMM, such as SULF1, A2M genes and P53 stability gene set, were screened out and confirmed through siRNA interference, mRNA sequencing and qRT-PCR.Conclusion This study suggests PAMM could serve as a useful marker of white adipogenic differentiation of ADSCs, but it exerts no evident effect on white adipogenic differentiation. The unveiled downstream genes and gene sets regulated by PAMM would provide new clues for the functional research of PAMM.]]> 2022/7/20 0:00:00 ZHANG Ming-Meng,WANG Ying,LIANG Jie,WU Hong-Fu,SHI Yu-Cang,WU Zhi-Yuan,RAO Min-La,PENG Jian-Yu,JIANG Zhi-Wen,LIU Xin-Guang and SUN Xue-Rong 210true 2 group IIA (sPLA2-IIA) plays an important role in the regulation of cellular lipid metabolism and signal transmission, and participates in a variety of acute and chronic inflammatory responses. Investigating the relationship between their dynamics, allostery and functions is of important significance.Methods The elastic network model (ENM), perturbation-response scanning (PRS) and protein structure network (PSN) methods are utilized to analyze the structural dynamics and allostery of 31 human sPLA2-IIA members, and explore the relationship between their shared/specific dynamics and functions.Results The results show that the catalytic residues and cysteine residues involved in disulfide bond formation, important for the enzyme’s catalysis and structural stability respectively, are of minimal mobility, which are the requirements for the enzyme’s shared functions; however, the 5 regions involved in the association with calcium ion/membrane are of high mobility, which embody the specificity of sPLA2-IIA members. Additionally, the PRS analysis reveals that the above five regions have a high sensitivity to external perturbations, suggesting their important roles in allosteric modulation, while those residues with a low sensitivity play an important role in maintaining structural stability. Finally, the ANM analysis indicates that the strong correlation movements around the catalytic sites of sPLA2, are helpful for the enzyme’s catalytic function exertion.Conclusion This study is helpful for the deep understanding of the dynamics and functionally allosteric mechanism of human sPLA2-IIA, and can provide a guide for drug design and accurate design of proteins with finely tuned activities.]]> 2022/7/20 0:00:00 ZHANG Shan,GONG Wei-Kang,ZHANG Na and LI Chun-Hua 209true Objective Allostery plays important roles in regulating protein biological functions. How to effectively identify the allosteric signal transduction pathway and the related key residues from the tertiary structure of proteins is a hot scientific problem in the research field of protein structure-function relationship.Methods In the present work, a method based on elastic network model (ENM) combined with force distribution calculation was used to investigate the response of proteins to the external loading forces, and then the allosteric pathway and associated key residues in proteins were identified based on the analysis of internal force distribution. In this method, external forces were exerted on the allosteric site of the protein, and then the deformations and internal force distributions within the protein in response to the external loading forces were analyzed. Based on these analyses, the key sites that are coupled with the deformation of the force-loading region were identified, and the transduction pathway of the force signals in the protein was obtained.Results By using the proposed method, two proteins, the human phosphoglycerate kinase (hPGK) and protein tyrosine phosphatase (PTP) PDZ2 domain, were investigated to identify the allosteric pathway and the related key residues in the systems. For hPGK, two allosteric pathways were identified, which mediate the transduction of force signals from the substrate binding site to the hinge region of the protein. For PTP PDZ2, two long-range allosteric pathways, from the ligand binding site to the distantly opposite side of the protein, were also successfully revealed. The calculation results are consistent with the experimental observations and the results obtained with molecular dynamics simulations.Conclusion This study provides an effective method for the identification of key residues and allosteric pathway in proteins.]]> 2022/7/20 0:00:00 LI Jiao,WANG Wei-Bu and SU Ji-Guo 208true Objective The prediction model based on PSSM (position-specific scoring matrix) has achieved good results, and various optimization methods based on PSSM are also being continuously developed. However, the accuracy rate is relatively lower. In order to further improve the prediction accuracy rate, this paper does further research based on the CNN algorithm.Methods In this paper, PSSM is used to process the letter sequence into a numeric matrix, and through a convolutional neural network (CNN) algorithm for classification. The 3 promoter sequences of Sigma38, Sigma54 and Sigma70 of E.coli K-12 (Escherichia coli K-12, hereinafter referred to as Escherichia coli) are used as the positive sets, and the sequences of the Coding and Non-coding regions of Escherichia coli are the negative set.Results In the prediction of Escherichia coli for the two-classification for promoters, the accuracy rate reaches 99%, and the success rate of promoter prediction is close to 100%; in the three-classification for Sigma38, Sigma54 and Sigma70 promoters, the prediction accuracy rate is 98%, and for each the prediction accuracy of these sequences can reach 0.98 or more. Finally, we tried 4 classifications of 3 promoters of Sigma38, Sigma54 and Sigma70 with Coding area or Non-coding area sequences respectively, the accuracy of prediction was 0.98. The prediction accuracy of the ten-fold cross-validation of the balanced samples of the Sigma promoters can reach more than 0.95, the Hamming distance is 0.016, and the Kappa coefficient is 0.97.Conclusion Compared with other classification algorithms such as SVM (support vector machine), the CNN classification algorithm has more advantages, and based on the classification advantages of CNN, the coding method can also be simplified.]]> 2022/7/20 0:00:00 PENG Bao-Cheng,ZHANG Xiao-Wei,LIU Yang and Fan Guo-Liang 207true Objective Male pattern baldness (MPB), or androgenetic alopecia (AGA), is a common type of hair loss in men, with an estimation that approximately 80% of the phenotypic variance can be explained by genetic factors. Most prediction models were developed in European and few MPB associated (single nucleotide polymorphisms,SNPs) have been validated in East Asian population. In this study, MPB associated SNPs in European were verified in Chinese population, and MPB risk prediction models were built based on those SNP data.Methods We examined 486 genetic variants previously reported associated with MPB, and assessed their impacts on hair loss in 312 Chinese individuals. Different sets of SNPs were selected by stepwise regression and Lasso regression. Logistic regression algorithm was used to construct the prediction models and the evaluations were conducted by the method of 10-fold cross validation. We further compared the prediction accuracy among logistic regression, k-nearest neighbor classifier, random forest and support vector machine.Results 174 SNPs demonstrated significant associations with MPB (P<0.05). Among those SNP markers, 22 SNPs and 25 SNPs were selected by different screening methods. Two logistic regression model considering the genotypes of 22 and 25 SNPs demonstrate that the risk of MPB were predictable at AUC (area under curve) level of 0.85 and 0.84. Prediction accuracy was slightly reduced after performing 10-fold cross validation, 0.81 and 0.77 respectively. Moreover, the AUC of both models reaches maximum (0.89) when age was added as a predictive factor. From the running results, the logistic regression prediction model had obvious advantages.Conclusion Overall, although the accuracy obtained here has not reached a clinically desired level, our model still has great potential for genetic prediction of MPB, which may assist decision making on early MPB intervention actions and in forensic investigations.]]> 2022/7/20 0:00:00 XUE Si-Yao,LI Cai-Xia,YUN Ke-Ming,CONG Bin and ZHAO Wen-Ting 206true Objective Bees are born with rich olfactory recognition capabilities. Foraging, mating, navigation and social activities all rely on their olfactory system. It is an ideal model for studying behaviors and neural mechanisms of olfactory perception, learning and memory. Bees can distinguish a compound odor as a configural character, and can distinguish the components individually as well, but yet it is not clear whether the feature component of a compound odor is stored into the memory as a key cue in a feature-dependent context.Methods In the feature-positive (FP: AB+, B-) and feature-negative (FN: AB-, B+) olfactory discrimination tasks, we train bees to learn to associate an odor and a sugar reward. During the mid-term memory (3 h) and long-term memory (24 h) tests, response to the trained odors AB and B, and the feature odor A were tested.Results We found that in the FP task, bees can form stable mid-term and long-term memories of the trained odors. The memory of the feature odor was well stored as the rewarded compound. In the FN task, bees were able to distinguish the two trained odors, but their response to the unrewarded compound increased with the passage of time.Conclusion Our results suggest that bees selectively consolidate the reward associated information into long-term memory no matter it is the compound or the components. Interestingly, the feature component is not the key factor to be consolidated into the memory system. Our study indicates that selective memory consolidation is supposed to be an important strategy for simple animals to efficiently encode survival-related information.]]> 2022/7/20 0:00:00 YAN Wei,MENG Zhi-Qiang and LIU Chang 205true Objective The T cell receptor-co-receptor complex (TCR-CD3) plays an important role in the adaptive immune response, and the interaction between its subunits has always been a research hotspot. Due to the limitations of traditional experiments, the study of transmembrane protein TCR-CD3 complexes cannot go deep into the microscopic level.Methods Therefore, we used molecular dynamics simulation methods to analyze the self-assembly mechanism of TCR-CD3 complexes.Results Through coarse-grained simulation (CGMD), we found that the TCR-CD3 complex has an assembly sequence in which αβ sequentially binds δε", γε, and ζζ" during the assembly process, and explained that the αR253 mutation reduces the interaction between αK258 and δε".Conclusion We demonstrated that only the transmembrane region of the TCR-CD3 complex is not sufficient to mediate the specific interaction between the subunits of the TCR-CD3 complex. The results of steered dynamics (SMD) and all-atom simulation (AAMD) indicate that the extracellular interaction between ζζ" and the rest of TCR-CD3 is stronger than the transmembrane region. The absence of ζζ" makes the stability of αβδε"γε little change, while the absence of δε" greatly reduces the stability of αβγεζζ".]]> 2022/6/21 0:00:00 WANG Feng-Li,CHEN Jia-Lin,HE Cheng-Zhi and LUO Shi-Zhong 204true Objective Magnetic seizure therapy (MST) is an emerging neuromodulation intervention technique that is widely used in the treatment of major depressive disorder (MDD), but its antidepressant mechanism is still unclear. Therefore, exploring the regulatory effect of MST on the brain functional network of MDD patients with different therapeutic effects is of great significance for the study of the antidepressant mechanism of MST treatment.Methods In this study, the resting-state EEG of 18 MDD patients pre- and post-MST was recorded, and the Pearson correlation method was used to construct the brain functional network; the complex network theory was used to compare and analyze the changes in the topology of the brain functional network; according to the rate of change of the HDRS-17 score, it was further distinguished. The network characteristics were performed a secondary comparison between responders and non-responders.Results The brain functional network connectivity, mean node degree, mean clustering coefficient, and mean global efficiency values were significantly higher than that pre-MST treatment. Mean path length values were significantly lower, and small world attribute significantly increased post-MST treatment. Compared with the group of non-responders, the changes in the characteristic parameters of the brain functional network in the group of responders were significantly larger.Conclusion MST significantly changes the topology of the brain functional network in patients with MDD, and it has a certain modulation effect on the brain functional network of patients. These results provide experimental support and a theoretical basis for the study of the antidepressant mechanism of MST treatment.]]> 2022/6/21 0:00:00 TIAN Shu-Xiang,XU Gui-Zhi,WANG Long-Long,LIU Xiang-Xiang and FITZGERALD B Paul 203true Objective The phenotypes of population, such as biogeographical and physical characteristics, are the result of the interaction between genotype and environment. A large number of genomic studies have shown that the Han Chinese are admixed population which displayed obvious genetic differences between the north and the south. The purpose of this study is to explore whether there are differences in epigenomes between northern and southern Han populations, and to screen biogeographical specific genetic loci.Methods Glint software was used to perform epigenome-wide association study (EWAS) analysis on genome-wide methylation microarray data of 483 Han Chinese individuals, and Lasso regression was used to screen the sites. Multiple logistic regression algorithm was used to construct the prediction model for the Han population in the south and the north, and the evaluation was conducted by the method of 10-fold cross validation.Results A group of CpG sites with significant differences between southern and northern Han populations were screened out, with accuracy of 99.03% and Kappa of 0.979 6.Conclusion This study shows that there are epigenetic differences between the southern and northern Han populations, which lays a foundation for further studies on epigenetic differences between the Han populations in different regions.]]> 2022/6/21 0:00:00 SUN Chang-Chun,XU JI-Chen,JIANG Li,GUO Xiao-Yuan,ZHAO Wen-Ting,YE Jian and LI Cai-Xia 202true Objective Current forensic SNP genotyping methods often require imported platforms and are labor-intensive, time-consuming and costly. Droplet digital PCR (ddPCR) is a new generation of PCR technology that allows rapid qualification of rare target DNA sequence, and is less susceptible to PCR inhibitors. This study is intended to establish a SNP genotyping method on domestic ddPCR platform and explore the applicability of ddPCR in forensics.Methods Genotyping system of the high altitude adaptive EPAS1 haplotype (rs115321619, rs73926263, rs73926264, rs73926265 and rs55981512) was established by ddPCR. Then we tested the specificity of primers and probes, evaluated its accuracy, stability, sensitivity and adaptability separately. Meanwhile, ddPCR and SNaPshot minisequencing technology were compared in terms of inhibitor resistance ability. Finally, preliminary application in 70 samples were conducted.Results The ddPCR assay only required a total run time within 2.5 h, and showed high accuracy and repeatability in SNP genotyping. The detection sensitivity was 0.312 5 ng. DdPCR assays also exhibited better tolerance to inhibitors than SNaPshot. The tested individuals showed good consistence with their background information.Conclusion The SNP genotyping assay based on ddPCR is accurate, rapid, easily-used and shows great resistance to perturbations by inhibitors, which has strong application potential in the field of rapid forensic detection, and is suitable for forensic analysis.]]> 2022/6/21 0:00:00 LIN Pei-Shuang,WANG Han,ZHU Ling-Xiang,BEI Lei,HUANG Jiang,ZHAO Wen-Ting and LI Cai-Xia 201true Objective TRIM28 (TRIpartate motif-containing protein 28), also called KAP1 (KRAB-associated protein1) or TIF1β (transcription intermediary factor 1β), is a heterochromatin related protein, composed of RBCC domain in N terminal and PHD,BROMO,NHD domain in C terminal, which participates in the establishment of H3K9me3 modification and SUMOylation pathway. The present study is to further explore the function of TRIM28.Methods We used CRISPR/cas9 technology, chromatin imm unoprecipitation assay, Western blot, quantitative real-time assay in HEK293F cell line to further study the function of TRIM28. We knocked out Trim28 gene in HEK293F cell line by CRISPR/cas9 technology.Results By analyzing experimental data, we found that TRIM28 mainly suppresses the transcription of genes with low endogenous expression level. Further analysis by GO bioinformation assay, we found that TRIM28 regulates zinc finger protein family genes and protocadherin protein family genes. According to the results of ChIP-seq, we found that in Trim28 knockout cell lines, the global distribution pattern of H3K9me3 decreased dramatically and H3K4me3 dropped moderately in TRIM28 target sites, indicating TRIM28 not only regulates the establishment of heterochromatin but also affects cis-acting elements. Meanwhile, H3K27ac modification, H3K4me1 modification and H3K4me3 modification in zinc finger protein family genes increased significantly, H3K9me3 decreased dramatically. H3K4me3 modification in protocadherin protein family genes increased significantly, H3K9me3 decreased dramatically. Previous study showed that protocadherin protein family genes mainly engaged in the identification and recognition of axon and dendrites in neuron cell.Conclusion In our study, it’s the first time to show the relationship between TRIM28 and protocadherin protein family genes. These results suggest that TRIM28 regulates zinc finger and protocadherin β genes by changing the chromatin accessibility and provide a new idea for further study of the relationship between TRIM28 and chromatin higher order structure.]]> 2022/6/21 0:00:00 ZHANG Xiang-Yu and BU Jia-Chen 200true Objective Opioid receptor, a kind of G protein-coupled receptors (GPCRs), mainly mediates an analgesic response via allosterically transducing the signal of endogenous ligand binding in the extracellular domain to couple to effector proteins in the intracellular domain. δ opioid receptor (DOP) is associated with emotional control besides pain control, which makes it an attractive therapeutic target. However, its allosteric mechanism and key residues responsible for structural stability and signal transmission are not completely clear. This paper aims to analyze the structural dynamics and allosteric effects of DOP.Methods Firstly, the relationships between DOP structure dynamics and function were explored by means of residue fluctuations in slow motion mode and fast motion mode from the anisotropic network model (ANM). Then, perturbation response scanning (PRS) was used to identify key residues related to allosteric communication in DOP.Results The DOP segments and functional sodium-binding sites can be well identified by the slowest motion modes, and the key residues that play a crucial role in protein structural stability can be identified by the fastest motion modes. Correlation analysis of residue motions reveals positive correlations between extracellular/intracellular transmembrane helices and loops, which promote the DOP structural stability and the binding of DOP with ligands. Key residues with high sensitivity and high effectiveness in PRS analysis play an important role in the allosteric communication of DOP.Conclusion This work sheds light on the allosteric communication mechanism of δ opioid receptor and provides valuable information for drug design.]]> 2022/6/21 0:00:00 CHEN Lei,GONG Wei-Kang and LI Chun-Hua 199true Objective Renal cell carcinoma is a malignant tumor originating from the renal tubular epithelial system. In the field of miRNA biomarkers for renal cancer, many previous researches had ignored the large gap in the amounts of samples between different subtypes of renal cancer, this may lead to differences in the diagnostic ability of selected miRNA biomarkers among patients with different subtypes of renal cancer, and may cause missed diagnosis and misdiagnosis. Therefore, we considered two subtypes of kidney cancer common markers for the study.Methods Statistics and two machine learning methods were performed to screen the expression profile data of clear renal cell carcinoma (ccRCC, KIRC) and papillary renal cell carcinoma (pRCC, KIRP) respectively and the results were intersected to obtain common miRNA markers for both types of kidney cancer. Then, ROC curve was used to verify the diagnostic ability of these biomarkers, machine learning methods using external data set (KICH) were also conformed to these biomarkers, the two methods further proved that these miRNA biomarkers’ diagnostic ability and avoided over-fitting. The rationality of these biomarkers was also verified by existing experimental literature. The molecular mechanisms of miRNA markers were investigated using bioinformatics methods.Results A total of 6 common miRNA markers for both types of kidney cancer were obtained (miR-21, mir-210, mir-185, mir-188, mir-362, mir-199a-2), 4 of them have been reported to be associated with renal cancer. Mir-188 and mir-199a-2 have not been reported to be associated with renal cancer, and maybe novel miRNA biomarkers of renal cancer. Then, we performed bioinformatic analysis on these 6 miRNA biomarkers, the results showed that the newly discovered biomarkers (mir-188 and mir-199a-2), were involved in the regulation of two renal cancer related pathway, MAPK signaling pathway and TGF-β signaling pathway. The differential expression of miRNA and its target genes in the pathway was verified, which further proved the reliability of miRNA as a marker and its regulatory effect on target genes. Also a possible mechanism of how 9 target genes of mir-185 (all belonging to the UGT1A gene family) participate in renal cancer was found, and there was no related literature.Conclusion The present study identifies possible new common miRNA markers for both types of kidney cancer and discovers a mechanism of kidney carcinogenesis that has not been seen in kidney cancer-related fields.]]> 2022/4/22 0:00:00 NIE Ren-Qing,TANG Tang,ZHANG Xiao-Yi and ZHANG Jin 198true Objective To explore the mechanism of heat shock factor (HSFl) alleviating coagulatory dysfunction in sepsis and protecting mice from acute lung injury.Methods In this study, a mouse model of sepsis was established by cecal ligation and puncture (CLP). We tested the coagulation indexes and pathological changes in the lungs of mice. Protein C expression was detected by ELISA, qRT-PCR and Western blotting. The expression level of protein C was observed by inhibiting or enhancing the expression of HSF1 by plasmid transfection, and the mechanism of HSF1 regulating protein C transcription was explored by bioinformatics, EMSA and dual luciferase reporter gene experiments.Results In the mouse model of sepsis, we discovered that the coagulatory activity of the HSF-/- mice was significantly enhanced and the lung injury was aggravated after CLP, compared with the HSF1+/+ mice. ELISA, qRT-PCR and Western blot showed that the expression level of protein C in the plasma and lung tissue of the HSF-/- mice was lower than that in the wild-type mice in sepsis. In vitro studies also demonstrated that HSF1 interference inhibited lipopolysaccharide (LPS)-induced protein C expression, while HSF1 overexpression enhanced protein C expression in bEnd.3 vascular endothelial cells. Further bioinformatics analysis indicated that the protein C promoter region contains HSF1 binding element (HSE). EMSA and dual luciferase reporter gene experiments showed that HSF1 bound to the HSE in the promoter region of protein C, thereby directly upregulating protein C transcription.Conclusion This study revealed that HSF1 was involved in acute lung injury in sepsis mouse model. HSF1 alleviated the coagulatory dysfunction in sepsis by directly upregulating protein C transcription, thus playing a protective role in mouse lung tissue.]]> 2022/4/22 0:00:00 WANG Hao,LI Tao,XIAO Gui,LIU Mei-Dong,LIU Ke,ZHANG Hua-Li,ZHU Ya-Xi and XIAO Xian-Zhong 197true Objective We investigated the role of glutathione transferase omega 1 (GSTO1) in regulating function of bone marrow-derived dendritic cells (BMDCs).Methods The expression of MHCII on the surface of Gsto1-/- BMDCs and GSTO1 inhibitor-treated BMDCs was detected by flow cytometry and confocal microscope. To investigate the role of BMDCs, they were cocultured with CD4 T cells from OTII mice in the presence of OVA and IFN-γ production was measured. The recycling and internalization of MHCII molecule in BMDCs treated with GSTO1 inhibitor were measured by flow cytometry. The transcriptional level of MHCII molecule in BMDCs treated by GSTO1 inhibitor was detected by real-time quantitative PCR. The total protein level of MHCII molecule in BMDCs treated by GSTO1 inhibitor was detected by Western blot. The ubiquitination level of MHCII after GSTO1 inhibitor treatment was detected by co-immunoprecipitation assay.Results GSTO1 deficiency and GSTO1 inhibitor did not affect proliferation and apoptosis of BMDCs, but reduced the surface expression of antigen presentation molecule MHCII. Gsto1-/- BMDCs and BMDCs treated with GSTO1 inhibitor showed impaired CD4 T cell-activating capability. In GSTO1 inhibitor-treated BMDCs, the recycling and internalization of MHCII molecule were normal. GSTO1 inhibitor showed no influence on the transcription of MHCII but reduced its total protein level. Furthermore, BMDCs treated with GSTO1 inhibitor showed higher level of ubiquitinated MHCII molecule and treatment with proteasomes inhibitor MG132 recovered the surface MHCII expression in these cells.Conclusion These data demonstrate that GSTO1 in BMDCs promotes MHCII expression and thus CD4 T cell-activating capacity by inhibiting ubiquitination of MHCII molecule.]]> 2022/4/22 0:00:00 LIU Ji-Wei,LI Shu-Hang,XIE Di,ZHANG Hui-Min,LI Qing and BAI Li 196true Objective Social cues such as eye gaze, head direction, and walking direction of biological motion are critical for human survival and social interaction. Since social and peripheral cues both have reflexive characteristics of attentional orientation, social attention is often regarded as one kind of exogenous attention. However, empirical evidence suggests that this explanation cannot fully account for all phenomena of social attention. Whether social attention and exogenous attention possess the same mechanism remains unclear.Methods Here, we used a typical spatial cueing paradigm to systematically examine the effects of cue validity on social attention and exogenous attention, triggered by eye gaze and peripheral cues, respectively.Results The results showed that both kinds of attention were affected by cue validity. With the increase of cue validity, the attention effects of eye gaze and peripheral cues increased. When the cue validity was noninformative (0.5) or strongly predictive (0.8), there was no significant difference in the attentional effects between social attention and exogenous attention. More importantly, however, when the cue validity was 0.2 (i.e., counterpredictive), the attentional effects of both cues were significantly different. While the facilitation effects of the eye gaze cue were weakened, the attentional effects of the peripheral cue were reversed and showed an inhibition pattern, suggesting that gaze-triggered attention is more strongly reflexive than exogenous attention orienting.Conclusion Our finding thus provides new evidence supporting the theoretical hypothesis that there exist significant differences between social attention and classical exogenous attention, at least in certain stages of their processing. Our study also offers a new method to distinguish social attention and exogenous attention through voluntary attentional control.]]> 2022/3/21 0:00:00 ZHANG Gui-Ting,YANG An-Min,SUN Jia-Lun,ZHOU Li-Qin and ZHOU Ke 195true Objective To evaluate the accuracy of predicting the kinship relationship between individuals based on the identity-by-state (IBS) algorithm.Methods The Illumina GSA chip was used to perform whole-genome detection on 253 samples. Based on high-density single nucleotide polymorphism (SNP) data, the IBS sharing statistics between two individuals was calculated to predict the kinship relationship. Filtering SNP by different conditional parameters to evaluate the influence of the number of sites on the accuracy of the algorithm’s prediction.Results The prediction accuracy rate of 1st-4th degree of relatives proved to be as high as 99%, with a paired difference of 1st degree and no false positive. Decrease in the number of SNPs has no significant impact on the accuracy of prediction, and the algorithm still achieves a higher accuracy rate even in the lower density of SNP markers.Conclusion The IBS algorithm provides an effective method for forensic genealogy inference, which has good application value for forensic on-site inspection materials with trace degradation.]]> 2022/3/21 0:00:00 GUAN Shan-Shan,ZHANG Wen-Jie,WEI Yi-Liang,LI Ying-Xiang,ZHAO Wen-Ting,FAN Hong and LIU Jing 194true Objective This study opens a new direction for photoacoustic tomography (PAT) to image human cervical trachea.Methods A multi-wavelength PAT experiment was performed on the cervical trachea of seven volunteers. The characteristics of high resolution (up to 150 μm) images of the respiratory cycle and different layers of tracheal wall were studied by PAT, and the imaging advantages and defects of this technique were evaluated.Results Our study demonstrated the ability of PAT to image human cervical trachea, and delineate the structural characteristics of it and its surrounding tissues. Variation of tracheal lumen at different phases of a respiratory cycle by PAT indicated its high time resolution. Collagen, as an endogenous contrast of PAT, was abundant in tracheal cartilage, which has a wavelength of absorption coefficient ranging from 900 nm to 1 300 nm. This study shows that the wavelength of 910 nm is best for tracheal cartilage imaging.Conclusion It can be considered as a non-invasive, convenient and non-ionizing radiation imaging method for real-time evaluation of human trachea. Given the high spatial resolution and real-time capabilities, PAT has the potential to become a useful tool for non-invasive airway imaging in future. We believe this study sets the ground work for the analysis of the human trachea using PAT in vivo.]]> 2022/3/21 0:00:00 WEN Yan-Ting,WU Dan,ZHANG Jing,GUO Dan,XIONG Chun-Yan,CHI Zi-Hui,CHEN Yi,LI Lun and JIANG Hua-Bei 193true Objective Lung cancer is one of the most commonly diagnosed cancers and the leading cause of cancer deaths worldwide. Despite significant advancements in combined therapy, the numbers of lung cancer cases and deaths continue to rise. Thus, novel therapeutic strategies are necessary to improve cure rates.Methods The effect of concomitant use of deferoxamine and chloroquine with frontline chemotherapy drugs was evaluated in lung cancer cells. The autophagy-dependent degradation of ferritin (i.e. ferritinophagy) was determined by western blot and immunofluorescence. The cell proliferation was quantified by cell member and cell cycle. The mitochondrial oxidative phosphorylation was determined by measuring the cellular oxygen consumption.Results The iron chelator deferoxamine induced ferritinophagy in cancer cells, and this process was blocked by chloroquine, an anti-malaria drug. Exposure of lung cancer cells to deferoxamine and chloroquine together inhibited mitochondrial oxidative phosphorylation, caused the accumulation of G1 phase cells and blocked cell proliferation. More importantly, the use of deferoxamine and chloroquine concomitantly with cisplatin or etoposide resulted in a significant increase in cytotoxicity in lung cancer cells when compared with the same concentration of cisplatin or etoposide alone.Conclusion These data indicate the efficacy of targeting ferritinophagy for the sensitization of iron-addicted cancer cells to chemotherapy, and introduce a potential targeted therapeutic approach for the treatment of cancer.]]> 2022/2/21 0:00:00 WANG Yong-Wei,XIAO Min,ZHANG Yu,HUO Yan-Wu and WEI Tao-Tao 192true Objective Lung cancer is one of the most common cancers in the world. Lung adenocarcinoma (LUAD) has the highest annual mortality rate among lung cancer patients. It has been reported that changes in gene spectrum were associated with the process of tumorigenesis and its development. The purpose of this study is to identify the gene signatures associated with LUAD and to further analyze their prognostic significance.Methods Weighted gene co-expression network analysis (WGCNA), differential gene analysis, cox regression analysis, and protein-protein interaction (PPI) network analysis were used to screen the hub genes highly related to LUAD based on The Cancer Genome Atlas (TCGA) database. The RNA-seq data sets from TCGA and GTEx (Genotype Tissue Expression) database were combined and divided into a training set and a validation set, which were used to construct the diagnostic model by support vector machine recursive feature elimination feature (SVM-RFE) algorithm. GSE32863 and GSE31210 were used to verify the diagnostic accuracy of the model and the prognostic value of our obtained gene signatures, respectively.Results The results demonstrated that the model of 5 gene signatures (anln, cenpa, plk1, tpx2, cdca3) obtained by the SVM-RFE algorithm had an outstanding performance in the classification of LUAD patients. Functional enrichment analysis showed that these 5 gene signatures were highly related to the biological process of tumor initiation and progression. What’s more, LUAD patients with high expression of these 5 genes also exerted a poor outcome in survival status.Conclusion Therefore, we could conclude that our study obtained useful models with 5 gene signatures for the diagnosis and prognosis of LUAD, which were essential for the development of novel targets applied in precision therapy.]]> 2022/2/21 0:00:00 WANG Mei,WANG Ke-Xin,TAN Jian-Jun and WANG Jing-Jing 191true Objective In order to analyze the number and morphology of cells in the process of cell culture conveniently.Methods In this paper, we introduce a cell counting method which can directly count cells in culture dish from images of commercial optical microscope by applying deep learning technology.Results In order to implement cell segmentation and counting, labeling and training is carried out on the image of adherent cells and suspension cells by a U-Net structure network. The cell growth curve is plotted and the inhibition rate of inhibitor is calculated by this algorithm, which shows the practicability of the algorithm.Conclusion It is feasible to do cell segmentation in dish by deep learning method.]]> 2022/2/21 0:00:00 JIA Ce,CAO Guang-Fu,WANG Xiao-Feng and ZHANG Xiang 190true Objective To investigated the function and the target gene of miR-216b in osteoclast differentiation and explored its effect on osteoclast cholesterol efflux.Methods The cell model of RAW264.7 osteoclast precursor cell differentiation induced by RANKL stimulation was established. Tartrate-resistant acid phosphatase (TRAP) staining assay was conducted to evaluate osteoclasts differentiation. MiR-216b target gene, ABCG1 3′ untranslated region (3'UTR) sequence and free energy were predicted and analyzed by bioinformatics analyses and dual-luciferase reporter assays. MiR-216b mimic or inhibitor transfection was performed to verify the role of miR-216b in osteoclast differentiation. Liquid scintillation counting was used to measure [3H]-labeled cholesterol efflux from RAW264.7 macrophage-derived osteoclasts. The lipid accumulation in RAW264.7 macrophages was detected by high performance liquid chromatography (HPLC). Real-time quantitative PCR (RT-qPCR) and Western blot assays were used to assess the transcriptional and post-transcriptional levels of ABCG1 in osteoclasts.Results Our results showed that the number of osteoclasts, the average diameter of osteoclasts and the fusion index were significantly increased when cells were transfected with miR-216b mimic, as revealed by tartrate-resistant acid phosphatase-positive staining and microscopy assay. MiR-216b inhibitor showed the complete opposite outcome which brought additional evidence to our findings. Bioinformatics analysis and dual-luciferase reporter assays showed that miR-216b targets the 3′UTR of ABCG1. Moreover, miR-216b suppressed both the mRNA and protein levels of ABCG1 in osteoclasts. Besides, we found that silencing of ABCG1 by ABCG1 siRNA increased the number of osteoclasts, the average diameter of osteoclasts and the fusion index. MiR-216b reduced cholesterol efflux from osteoclasts by inhibiting ABCG1 expression.Conclusion Collectively, these findings suggest that miR-216b downregulates ABCG1 expression and inhibits osteoclast cholesterol efflux, which disturbs cholesterol homeostasis and promotes osteoclastogenesis.]]> 2022/2/21 0:00:00 YANG Wu-Zhou,LI Heng,YU Xiao-Hua,ZHANG Jie,HUANG Xin-Yun,ZHAO Zhen-Wang,CAO Qi and TANG Chao-Ke 189true Objective To investigate the effect of two indomethacin preparations on the healing process of oral ulcers.Methods After the right cheek of rats were burned with nails,they were divided into indomethacin gel to medicine group (YN), compound benzocaine gel positive control group (NP), indomethacin spray to medicine group (YP), stomatitis spray, normal control group (PP), positive control group (N) and the negative control group (M). The reepithelialization of ulcer surface and granulation tissue formation was evaluated after HE staining with histological quantitative standard evaluation. The level of nitric oxide (NO) and nitric oxide synthase (NOS) in ulcer tissues was detected by colorimetry. The expressions of epidermal growth factor (EGF), as well as its receptor (EGFR) and vascular endothelial cell growth factor (VEGF) in rat ulcer tissues, were detected by immunohistochemistry.Results The ulcer area of the YP, YN, NP, and PP groups was smaller than that of the M group. The quantitative histological evaluation showed that the scores of the YP, YN, NP, and PP groups were higher than that of the M group (P<0.05). After administration, the level of NO and NOS in the YP group were significantly lower than that of M group (P<0.05). The EGF expression of YP, YN, NP, and PP groups was higher than that of the M and N groups (P<0.05). EGFR was expressed in the epithelium from the cytoplasm to the membrane, and the expression level in the YP, YN, NP, and PP was significantly higher than that of the N group (P<0.05). The VEGF expression of YP, YN, NP, and PP groups was lower than that of the M group (P<0.05).Conclusion Both two indomethacin preparations can reduce the inflammatory response and promote the healing of oral ulcers in rats.]]> 2022/2/21 0:00:00 ZHANG Shao-Lu,LUO Shu-Yu,QIU Yu-Ling,ZHANG Xiang-Yu,ZHONG Yu-Xu and KONG De-Xin 188true Objective To rescue a novel recombinant oncolytic influenza virus carrying OX40 ligand and evaluate its oncolytic efficacy in hepatocellular carcinoma.Methods The gene of encoding OX40L was optimized and inserted into the NS fragment of influenza virus A/PuertoRico/8/34 (PR8). Then, the recombinant plasmid along with the remaining 7 plasmids pHW191-PB2, pHW192-PB1, pHW193-PA, pHW194-HA, pHW195-NP, pHW196-NA and pHW197-M of influenza virus PR8, were co-transfected into COSⅠ/ MDCK cells. A recombinant oncolytic influenza virus, named rFlu-OX40L, was successfully generated using reverse genetics (RG). The titers of rFlu-OX40L were determined with hemagglutination and TCID50 methods. We observed the morphology and size distribution of rFlu-OX40L under electron microscopy. The growth curve of rFlu-OX40L was detected on MDCK cells. Cell viability of rFlu-OX40L was examined on various hepatoma cells with MTS assay. Cell death patterns induced by the rFlu-OX40L virus was analyzed with flow cytometry. The antitumor effect of rFlu-OX40L was evaluated based on the hepatocellular carcinoma tumor-bearing mice model.Results The oncolytic influenza virus rFlu-OX40L could be stably at the HA titer of 27－8 after passage in chicken embryos. The viral titers were 7-8 LgTCID50 /ml. The growth curve of rFlu-OX40L was consistent with that of wild-type PR8 virus and reached the peak values at 72 h. MTS assays revealed that rFlu-OX40L at dose of 3 MOI could significantly reduce the cell viability on HCC cells, without destroying normal cells in a time and dose-dependent manner. Flow cytometry results displayed rFlu-OX40L at dose of 3 MOI could induce cell apoptosis on HCC cells in a time- and dose-dependent manner. Compared with PR8 and PBS group, the CD3+, CD4+, CD8+, CD45+, CD69+ T cell numbers in spleen cells of mice inoculated with rFlu-OX40L increased significantly.Conclusion The recombinant oncolytic influenza virus rFlu-OX40L carrying OX40L could selectively destroy hepatocellular carcinoma cells in vivo and in vitro, which will provide a novel immunotherapy strategy for clinical management of hepatocellular carcinoma.]]> 2022/12/20 0:00:00 YANG Hao,DENG Zhuo-Ya,SUN Fang,LEI Guang-Lin,CHENG Jin-Xia,YU Hong-Yu,TIAN Chong-Yu,ZHANG Shao-Geng and YANG Peng-Hui 187true Objective Hepatic stellate cells (HSCs) are the main producers of fibrotic extracellular matrix (ECM) and play a critical role in the initiation, progression, and regression of hepatic fibrosis (HF). Dihydromyricetin (DMY) has hepatoprotective properties, but the mechanism of this protection is unclear. Our study examined the effects of DMY on the activation of HSCs triggered by ferric ammonium citrate (FAC) in HSC-T6 cells and explored the possible mechanisms of the hepatoprotective properties of DMY.Methods Cell viability was evaluated using MTT assay. The levels of ECM in the culture supernatant were examined usingenzyme linked immunosorbent assay. The iron deposition levels in HSC-T6 cells were assessed using Prussian Blue staining. The total iron and free iron levels in HSC-T6 cells were measured using colorimetric assay and calcein-AM assay, respectively. The ultrastructure of HSC-T6 cells was observed using transmission electron microscopy. The expression levels of ferritin heavy chain 1 (FTH1), α-smooth muscle actin (α-SMA), nuclear receptor coactivator 4 (NCOA4), microtubule-associated protein 1 light chain 3(LC3), and p62/SQSTM1 were measured using Western blotting.Results Compared with the FAC group, the DMY+FAC group had a significant reduction in the main components of ECM, total iron and free ironlevels, expression levels of α-SMA, NCOA4, and LC3-Ⅱ proteins, and the ratio of LC3-Ⅱ/LC3-Ⅰ; there was also a significant upregulation of the protein expression levels of FTH1 and p62. Furthermore, rapamycin partially blocked the effects of DMY, which inhibited the activation of HSCs induced by FAC.Conclusion DMY inhibits the activation of HSCs induced by iron overload, and the underlying mechanism may be involved in the inhibition of ferritinophagy.]]> 2022/12/20 0:00:00 ZENG Bin,ZHOU Shou-Hong,XU Zi-Wei,CHU Yu-Yang and DUAN Wu-Xia 186true Objective The stable isotopic composition of human tissue is related to the individual’s diet, environment and metabolic status. Once grown, human hair is no longer exchanged with the body and is chemically stable and easy to collect, making it a good subject for studying the stable isotope composition of human tissue. The oxygen and hydrogen elements that make up the human body are mainly derived from the water and food, of which the oxygen and hydrogen stable isotope composition is recorded in hair keratin. Differences in the stable isotope composition of oxygen and hydrogen in the human hair of different regions can be used to infer information about people’s diets, living areas and activity trajectories, which is of great importance in forensic science and other research fields.Methods In this study, oxygen and hydrogen stable isotope ratios of hair samples from permanent residents in different regions of China were detected and analyzed using an elemental analyzer-isotope ratio mass spectrometer (EA-IRMS).Results The results showed that there were significant differences in δ18O and δ2H between some cities, and a significant positive correlation between overall δ18O and δ2H. The overall discrimination accuracy of the stable isotope data was 63.9% for the cross-validation, and increased significantly when combined with the carbon and nitrogen stable isotope data, reaching an overall discrimination accuracy of 76.0% for the cross-validation. As the type of stable isotopes used in the discriminant analysis increased, the model’s discriminant ability was significantly enhanced.Conclusion The overall discriminative accuracy of the multi-layer perceptron neural network model built by using the stable isotope data of the 4 elements was 82.8%, and the overall discriminative accuracy of the radial basis function neural network model was 78.8%. The multilayer perceptron neural network model had the highest discriminative accuracy among the three traceability mathematical models.]]> 2022/12/20 0:00:00 YU Zi-Yang,YANG Rui-Qin,HUANG Yang,QIN Hao,HU Can,ZHU Jun,WANG Ping,GUO Hong-Ling and MEI Hong-Cheng 185true Objective In recent years, neuromodulation techniques for brain function regulation have developed vigorously, and many methods have been popularized and applied in clinical practice, mainly including electrode deep brain stimulation, transcranial magnetic stimulation, photogenetic technology, ultrasonic deep brain stimulation, etc. However, these regulatory techniques have some problems, such as poor flexibility of stimulating target change, insufficient spatial resolution, and need to inject virus transfection. Compared with these techniques, terahertz (THz) wave regulation can intervene neural activity with higher spatial and temporal resolution without introducing foreign genes. Laser nerve stimulation is a specifically targeted stimulation method, which can control the excitation or inhibition of nerve by adjusting different laser parameters (laser wavelength, pulse energy, etc.). However, due to the lack of experimental means and platforms in this research direction, few relevant studies have been carried out.Methods To sum up, this paper sets up different testing platforms for related researches at molecular, cellular and somatic levels, starting from auditory nerve.Results The experimental results show that these systems have good coupling and targeting properties in time and space, and the measured signals are less disturbed by noise.Conclusion These systems can effectively test the response to terahertz stimulation and precisely control the timing and location of the stimulus.]]> 2022/12/20 0:00:00 TAN Xiao-Xuan,YANG Xiao,CHEN Shuo,YIN Jun-Kai,FENG Yi-Fei,YUAN Yi-Fang and WU Kai-Jie 184true Objective The present study attempted to design novel miRNA expression cassettes (MEC) targeting specific consensus sequences of hTERT and hTR, which remedied the degradability and cytotoxicity and provided a convenient method for miRNA synthesis.Methods The MECs specific to hTERT and hTR were constructed by overlap polymerase chain reaction (PCR). Telomeric repeat amplification protocol (TRAP)-silver staining and TRAP real-time PCR analysis were used to determine the telomerase activity. The telomere length was determined by real-time PCR, whereas cell viability was determined using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The cell apoptosis rate and cell cycle were assessed using the annexin V/propidium iodide (PI) double staining and PI single staining assays, respectively, coupled with flow cytometry.Results The telomerase-specific MECs were successfully constructed. Each MEC inhibited the telomerase activity differently. Telomerase silencing could induce immediate growth arrest in the G0/G1 phase and led to retinoblastoma (RB) cell apoptosis.Conclusion miRNA-mediated telomerase silencing is an efficient strategy to impair RB cell growth. A robust system must be developed to fully explore the efficacy of miRNAs. The constructed MECs exhibited a strong RNAi effect and thus may be utilized to effectively screen RNAi-targeted sequences for RB gene therapy.]]> 2022/12/20 0:00:00 ZHAO Dan-Dan,DING Cai-Yun and LI Wen-Tao 183true Objective To investigated the N-linked glycans in bladder cancer FFPE tissue sections, and explored abnormal N-linked glycans modification in bladder cancer FFPE tumor tissues sections.Methods An experimental procedure for in situ extraction of N-linked glycans based on FFPE tissue sections was developed. FFPE tissue sections were digested by PNGase F and liberated N-linked glycans. Permethylation was performed to modify the free end of the N-linked glycans. The relative intensity of N-linked glycans was detected by MALDI-TOF/TOF-MS. Database matching was performed to determine possible glycoforms of N-linked glycans. The accuracy of significantly different N-linked glycans as a predictive bladder cancer biomarker were predicted by ROC analysis.Results The data of MALDI-TOF/TOF-MS detection of premethylation-modified N-linked glycans showed that, in tumor and peritumoral FFPE tissue sections of 16 patients with bladder cancer, the relative intensity of N-linked glycans N2H6, N2H7, N2H8, N2H9 (high mannose) and N5H6F1 (complex) were increased significantly in the tumor tissues of bladder cancer. At the same time, N2H5 (high mannose), N3H5(hybrid) and type N3H4, N4H4, N5H6F1S2 (complex) N-linked glycans were significantly decreased. ROC analysis showed that the biantennary N-linked glycan N3H4 (AUC=0.90) and N4H4 (AUC= 0.91) were reliable in distinguishing tumor and peritumoral tissue of bladder cancer patients separately or combined together, and these N-linked glycans may become a potential biomarker for bladder cancer.Conclusion Abnormal N-glycosylation of proteins in bladder cancer FFPE tumor tissue, N-linked glycans N3H4 and N4H4 may be potential biomarkers of bladder cancer.]]> 2022/10/21 0:00:00 CHENG Ying,SUN Cheng-Wen,QIN Yan,SHI Shuai,LI Yue-Yang,FAN Qi-Gao,YANG Gang-Long and GAO Xiao-Dong 182true Objective Molecular imaging technology has the characteristic of “early detection”, since aberrations at molecular levels occur much earlier than the changes at the anatomical level. In this study, intracellular photoacoustic molecular imaging (PMI) method was used to map the folic-acid-conjugated gold nanorods (FA-AuNRs) targeted on cancer cells.Methods We synthesized the FA-AuNRs and studied their properties including morphology, absorption spectrum, and biocompatibility. The conjugated folic acid has endowed the FA-AuNRs with the ability to target folate-receptor-expressing cancer cells. Then, PMI experiment was performed to study the target specificity of FA-AuNRs on cancer cells.Results FA-AuNRs were rod-shaped with a near-infrared absorption peak at ~800 nm. We observed the strong photoacoustic signals in the cytoplasm of cancer cells, while weak photoacoustic signals in the normal cells, indicating the selective uptake of FA-AuNRs inside cancer cells via folate-receptor-mediated endocytosis. The study demonstrated the ability of PMI to precisely map the FA-AuNRs targeted on cancer cells.Conclusion With the aid of specific targeting, information about the surface molecules of cancer cells can be obtained by PMI. This method is expected to visualize, characterize and quantify biological processes at cellular and molecular levels.]]> 2022/10/21 0:00:00 ZHONG Jun-Ping and CHEN Jin-Ying 181true Objective To investigate the toxic effect and molecular mechanism of silicon dioxide nanoparticles (SiO2NPs) on mouse Sertoli cells (TM4), TM4 cells were exposed to medium containing various concentrations of SiO2NPs (0, 1, 10, 100 mg/L) for 24 h.Methods After treatment, the morphology and viability of TM4 cells were detected by optical microscope and cell counting kit (CCK-8). The level of intracellular reactive oxygen species (ROS) was measured by fluorescent probe DCFH-DA, and the content of malondialdehyde (MDA) and activity of superoxide dismutase (SOD) were detected by kits according to the manufacturers’ protocol. The percentage of apoptotic cells in TM4 cells was detected by Annexin V-FITC/PI kit, and the expression levels of apoptotic molecules including Fas, FasL, Caspase-8, Caspase-3, Bax and Bcl-2 were detected by Western blot.Results The results showed that SiO2NPs inhibited the cell survival rates, decreased cell concentrations and changed cell morphology of TM4 cells in a dose-dependent manner. In addition, the level of ROS in TM4 cells was significantly up-regulated after exposure, followed by an increase in the content of MDA and the activity of SOD. Further study found that SiO2NPs increased cell apoptosis rate and activated the pro-apoptotic signaling pathway mediated by Fas/FasL. Interestingly, inhibition of oxidative stress by NAC in TM4 cells can alleviate the cell injure and apoptosis induced by SiO2NPs.Conclusion In summary, SiO2NPs induce TM4 cells apoptosis through oxidative stress and activation of Fas/FasL signaling pathway.]]> 2022/10/21 0:00:00 WANG Xu-Ying,GUO Zhi-Yi,HAO Hui-Yu,SUN Fan-Li,ZHANG Pin-Zheng,MENG Fang-Yu,CHEN Zi-Yun,LI Jin-Ze and SHANG Xuan 180true Objective To prepare and evaluate the biocompatibility of placental acellular matrix and to explore its feasibility as tissue repair material.Methods The waste placental tissue from delivery was handled by virus inactivation, acellular treatment and freeze-drying to obtain the acellular spongy matrix material of placenta. The effect of acellular was observed by HE staining and the microstructure of the materials was observed by scanning electron microscope. At the same time, 39 healthy male SD rats with body mass ranging from 120 g to 150 g were selected, randomly divided into experiment 1 group, experiment 2 group and control group. The subcutaneous implantation of the constructed matrix material was performed in rats, experimental group 1 implanted matrix materials, experimental group 2 implanted matrix materials and umbilical cord mesenchymal stem cells, the control group was the sham operation group. Routine blood tests were performed on the 3rd, 5th and 7th day after surgery to analyze the number of lymphocytes, granulocytes and other inflammatory cells. At 1, 2, 4, 8 and 9 week after the operation, the tissue samples of the implanted material and surrounding tissues were taken for HE staining analysis.Results The constructed acellular matrix of placenta appears milky white spongy in macroscopic view, HE staining indicated no cell residue. Under electron microscope, the internal void of the material was obvious, and the crosslinking degree of the material was good. The total porosity was (77.54±2.53)%. After subcutaneous implantation, the incision healed well, and no obvious inflammatory cell increase was observed in blood routine examination. Seven days after the operation, blood vessel formation can be observed by HE staining in the sections of the implanted materials. Moreover, the addition of umbilical cord mesenchymal stem cells can accelerate the fusion of the materials with the body, promote the in-depth growth of cells and vascularization.Conclusion Placental acellular sponge matrix material has good biocompatibility and can be an ideal source of tissue engineering materials.]]> 2022/10/21 0:00:00 LIU Xiu,ZANG Chuan-Bao,LIN Yan-Chao,CHENG Long and FU Xiao-Meng 179true Objective Although many improvements in soybean transformation have been made, the efficiency of transformation is still low. Thus, a more useful and efficient soybean transformation system required to be established. The objective of this study is to develop high frequency induction and transformation of somatic embryos (SEs) using mature seed-derived leaf (MSDL) of soybean as target explants.Methods The leaves detached from 7-d-old germinated seedlings were divided into 1.2 cm×1.2 cm sections. These sections were used to induce somatic embryogenesis on the embryo induction medium (EIM) and to infect with Agrobacterium EHA105 strain harboring the pCAMBIA1301 for transformation.Results The MSDL including primary leaf and secondary leaf was able to produce SEs. Most of SEs occurred along the incision of leaves. This system was shown to be applicable to different genotypes. The highest induction frequency of SEs was 95.0% and the average transformation efficiency based on β-glucuronidase (GUS) assay reached to 75.4%. The transgenic events were further con?rmed by Southern blot.Conclusion High frequency induction and transformation of somatic embryos using soybean MSDL as target tissue were developed in this study. This system can be applied to the development of genome editing technology in soybean.]]> 2023/8/14 11:14:27 SUN Ming-Jie,HOU Heng,ZHAO Chun-Mei and XUE Ren-Gao 178true Objective The study aims to explore the role and molecular mechanism of circPVT1 in invasion and migration of nasopharyngeal carcinoma (NPC), a malignant tumor originating from nasopharyngeal epithelium with the clinical feature of lymph node metastasis. circPVT1, a circular RNA formed by back splicing of exon 2 of PVT1 gene, has been found to be upregulated in various tumors and plays an important role in tumorigenesis. The study investigated the role and molecular mechanism of circPVT1 in invasion and migration of NPC cells.Methods We detected the expression of circPVT1 and its downstream miRNAs and FSCN1 in NPC cells using RT-qPCR, and evaluated the effect of circPVT1 on NPC cell invasion and migration using Transwell and scratch healing experiments. RNA pull-down experiment was used to detect the miRNAs bound by circPVT1, and the dual luciferase reporter assay was used to detect the targeting of FSCN1 mRNA by miR-24-3p and let-7a-5p.Results The study found that overexpression of circPVT1 in NPC cells can promote invasion and migration, while knockdown of circPVT1 can inhibit NPC cells invasion and migration. Further studies revealed that circPVT1 upregulates the expression of FSCN1 by competitively adsorbing miR-24-3p and let-7a-5p.Conclusion circPVT1 promotes invasion and migration of NPC cells through the miR-24-3p/let-7a-5p/FSCN1 axis, and is an important driving factor in the development of NPC.]]> 2023/8/14 11:14:38 MO Yong-Zhen,WANG Yu-Min,FAN Chun-Mei,YAN Qi-Jia,ZENG Zhao-Yang,JIANG Wei-Hong and XIONG Fang 177true Objective The effects of zirconium dioxide nanoparticles (ZrO2-NPs) exposure on the histone H3 modification in human skin keratinocytes (HaCaT) were clarified, and the underlying mechanism of histone H3 modification changes was explored in this study to provide a theoretical basis for the further safe application of nanomaterials.Methods The ZrO2-NPs were firstly characterized in detail by means of scanning electron microscope, laser particle size analyzer, X-rad diffraction, etc. Subsequently, the effects of ZrO2-NPs exposure on the cell viability, intracellular accumulation and histone H3 modification were evaluated by Western blotting and flow cytometry.Results The results showed that ZrO2-NPs after dispersion treatment exhibited obvious agglomeration, and their specific surface area decreased whereas the secondary particle size increased. The phosphorylation of histone H3 at serine 10, the acetylation of histone H3 at lysine 9 and 14, the trimethylation of histone H3 at lysine 4 and 27 were induced to upregulate by ZrO2-NPs in a short time (1 h). Through further analysis, it was found that the intracellular accumulation of ZrO2-NPs and the level of DNA damage caused by ZrO2-NPs were linearly correlated with the level of histone H3 modification induced by ZrO2-NPs.Conclusion These results suggested that the changes of the common modification sites of histone H3 in HaCaT cells were induced after exposure of ZrO2-NPs. The intracellular accumulation of ZrO2-NPs is one of the key factors in its induction of changes in histone H3 modification, and regulation mechanism of histone H3 modification may be involved in DNA damage repair pathways.]]> 2023/8/14 11:14:50 ZHAO Xiao-Xu,YAO Yan-Fang and HOU Qiao-Li 176true Objective Phosphatidylinositol 3-kinase gamma (PI3Kγ) plays a critical role in the immune system, thus identifying PI3Kγ as a potential therapeutic target for inflammation. However, there are few reports of selective PI3Kγ inhibitors being used in the field of anti-inflammation thus far, therefore, great efforts would be needed to study the anti-inflammatory effect of PI3Kγ inhibitors. In the present study, Eganelisib, the PI3Kγ inhibitor in clinical studies, was chosen to investigate the anti-inflammation and PI3Kγ inhibitory preference.Methods Firstly, lipopolysaccharide (LPS)-induced RAW264.7 macrophage was used as in vitro inflammation model to estimate the anti-inflammation of Eganelisib. Then, a modeling strategy integrated common feature pharmacophore, receptor-ligand pharmacophore, and molecular dynamics simulation was employed to reveal the γ-selective mechanism of Eganelisib.Results Eganelisib treatment could downregulate the production of inflammatory mediators by suppressing the PI3K signaling pathway. The common feature and receptor-ligand pharmacophore models found that the hydrogen bond and hydrophobic features play key roles in the interaction between Eganelisib and PI3Kγ protein. Several key residues such as Val882, Met804, Trp812, and Ile963 contributing to favorable PI3Kγ binding were highlighted in molecular dynamics simulation.Conclusion Our findings provide the initial anti-inflammatory effect of Eganelisib, and reveal the mechanism of selective binding between inhibitors and PI3Kγ, which may provide some guidelines for the development of anti-inflammatory PI3Kγ inhibitors.]]> 2023/8/14 0:00:00 XIONG Wen-Dian,JIA Lei,CAI Yan-Fei,CHEN Yun,JIN Jian and ZHU Jing-Yu 175true Objective Magnetically-coupled resonant wireless power transfer (MCR-WPT) has the advantages of large transmission distance, high transmission efficiency and penetration, which brings convenience to people’s lives. However, the safety of bioelectromagnetic effects caused by MCR-WPT has attracted much attention. In this paper, the influence of MCR-WPT electromagnetic environment on the characteristics of neuronal K+ channels in the dentate gyrus (DG) region of the mouse hippocampus was studied, in order to provide an experimental basis for the development and rational development and application of WPT technology.Methods Mice were continuously irradiated for 5 h for 30 d, and the learning and memory ability, instantaneous outward K+ channel current (IA) and delayed rectification K+ channel current (IK) of neurons in the hippocampal DG region were compared and analyzed by the control group and the magnetic field exposure groups of 5 d, 15 d and 30 d.Results The activation process of transient outward potassium channels in the magnetic field exposure group was inhibited, and the activation characteristics of delayed rectified potassium channels shifted to the depolarization direction, which reduced the outflow of K+ and enhanced the nerve excitability, but there was no significant change in the behavior of the magnetic field exposed group mice.Conclusion Long-term exposure to the electromagnetic environment of MCR-WPT altered the I-V properties and kinetic properties of K+ ion channels, inhibited IA and IK, and changed the frequency of neuronal action potential issuance, but these changes did not cause a decrease in learning memory capacity and cognitive dysfunction in mice.]]> 2023/8/14 11:15:40 ZHAO Jun,ZHANG Bing-Qian,ZHAO Yi-Hang and WANG Xiao-Xuan 174true Objective Chitosan (CS) is a natural broad-spectrum antibacterial active substance. Previous studies revealed that the interaction between chitosan and bacterial cell membrane play a key role in its antibacterial activity. However, Due to the limited characterizing capability of current experimental techniques, the exact mechanism of the interaction between chitosan and bacterial cell membranes remains to be studied. This paper aims to study the molecular mechanism of the interaction between chitosan and bacterial cell membranes.Methods In this study, all-atom molecular dynamics simulations were used to explore the dynamic interaction between totally deacetylated chitosan in different degrees of polymerization (8-, 12-, and 16-saccharides) with the outer membrane (OM) of Gram-negative bacteria and the cytoplasmic membrane (CM) of Gram-positive bacteria.Results The amino groups, carbon 6 hydroxyl groups and carbon 3 hydroxyl groups of chitosan play the determinant role in the initial attachment to the polar headgroup regions of the OM and CM. Interestingly, the terminal glycosyl units of chitosan inserted into the OM with an averaged depth of 1 nm where the sugar formed stable hydrogen bonds with the carbonyl groups on the fatty acid tails of the OM lipid A molecules. In contrast, chitosan could not insert into the CM steadily. Further, we found that the binding of chitosan to both OM and CM reduced their area per lipid, displacing the Ca2+ and Na+ from the headgroup regions of the membranes and thereby attenuating the cation-mediated interactions between membrane lipids.Conclusion Our results demonstrated that the positively charged amino groups of chitosan are essential for the interaction with bacterial membranes, which reduced the interlipid interactions and lead to the structural disorganization of bacterial membranes. These information provides new insights into the antimicrobial mechanism of chitosan at the atomic level.]]> 2023/8/14 0:00:00 ZHU Jing-Yi,MA Zhen-Yu,XIAO Min,WANG Lu-Shan and JIANG Xu-Kai 173true Objective The unclear mechanism of depression occurrence and the undesired clinical efficacy of drugs are the difficult problems all over the world. Previous studies have found that gaseous formaldehyde (FA) exposure or intraperitoneal injection of FA can directly induce depression-like behaviors in mice. Whether endogenous FA leads to depression is unclear. This study was to explore whether lipopolysaccharide (LPS) induces depression-like behaviors in mice by stimulating endogenous FA generation, and to observe whether non-invasive physical therapy——630 nm red light irradiation can activate FA dehydrogenase and degrade FA, thereby attenuate depression-like behaviors in mice.Methods Male adult C57BL/6J mice were randomly divided into: (1) control group, intraperitoneal injection of phosphate buffered saline (PBS); (2) depression model group, intraperitoneal injection of LPS at a concentration gradient; (3) red light intervention group, these mice were intraperitoneally injected with LPS and treated with 630 nm red light irradiation. The depression-like behaviors of mice were evaluated by open field test, sucrose preference test, tail suspension test and forced swimming test. The concentration and distribution of FA in brain tissue of mice were detected by FA fluorescence (Na-FA, specific FA fluorescent probe) quantitative method and whole brain FA fluorescence imaging. The concentrations of IL-1β, TNF-α, IL-6 and semicarbazide-sensitive amine oxidase (SSAO) were detected by ELISA.Results One hour after acute injection of LPS, FA accumulated rapidly in the whole brain, especially in the midbrain. With the increase of IL-1β and TNF-α content; 24-hour depression-like behaviors, for example, open field total movement distance, central area movement time (distance) decreased significantly; the total immobility time of tail suspension test and forced swimming test were increased. However, 630 nm red light treatment significantly attenuated LPS-induced depression-like behavior in mice accompanied by a decrease in midbrain inflammatory factors IL-1β and TNF-α.Conclusion The accumulation of FA in the midbrain is an initiating factor for depression, stimulating a large release of inflammatory factors and leading to depression-like behaviors. However, red light irradiation can scavenge FA and reduce neuroinflammation. It is a promising new strategy for non-invasive treatment of depression.]]> 2023/7/19 12:03:26 ZHAO Dan-Rui,ZHANG Feng-Ji,LIU Yi-Ying,WU Yi-Qing,ZHAO Hang,LIAO Hai-Kang,PAN Wen-Hao,CHENG Jian-Hua and TONG Zhi-Qian 172true Objective To investigated the effects of obesity on the structure of seminiferous tubules, autophagy and apoptosis-related proteins in testes tissue, and the effect of exercise on autophagy and apoptosis in testis and regulatory mechanism.Methods Fifty 6-week-old SD male rats were randomly divided into the standard diet group (SD, n=20) and the high-fat diet group (HFD, n=30). HFD group was fed 8 weeks to establish an obesity model of rats, and 20 rats were randomly selected to exercise intervention. Rats in SD and HFD groups were randomly divided into normal diet control group (CC), normal diet+exercise group (CE), high-fat diet control group (OC), and high-fat diet+exercise group (OE), 10 rats in each group. The rats in the CE and OE groups underwent moderate intensity treadmill training for 60 min/d, 5 d/week. CC and OC groups maintained the same feeding conditions. After 48 h of the last exercise, the rats were anesthetized intraperitoneally and weighed, the testes were taken from both sides of the rats, weighed and the testicular index was calculated. Paraffin sections of the testes were prepared to discover the testicular tissue structure by HE staining. Western blot was used to detect the protein expression of p62, LC3II, LC3I, BCL-2, Bax, and AMPK in testicular tissue, and the ratio of LC3II/LC3I was calculated. Immunofluorescence was used to detect LC3 and BCL-2 protein expression positions in the testis.Results The testicular index was reduced; HE staining revealed a significant decrease in the diameter of the seminiferous tubules (P<0.01), spermatocytes, and lipid droplet deposition in the testicular tissue; the protein expression of p62 and Bax were significantly increased (P<0.01), AMPK, LC3II/LC3I ratio, and BCL-2 were significantly decreased (P<0.01), and the number of LC3 proteins on sperm cells were significantly reduced in the testis tissue of rats in the OC group compared with the CC group. After exercise intervention, the testicular index was increased in the OE group; HE staining showed that the diameter of seminiferous tubules (P<0.01) and the number of spermatogonia (P<0.05) were significantly increased, and the structure of seminiferous tubules was improved; the protein expression of p62 and Bax were significantly decreased (P<0.01), AMPK, LC3II/LC3I ratio, and BCL-2 were significantly increased (P<0.01); the level of BCL-2 proteins on spermatocytes and sperm cells increased significantly in the OE group compared with OC group. AMPK was strongly correlated with p62, LC3II/LC3I, and Bax in all rat testicular tissue.Conclusion High-fat diet-induced obesity inhibits autophagy and promotes apoptosis in rat testes; 8 weeks of moderate-intensity exercise may activate autophagy and inhibit apoptosis in testis tissue via AMPK, correcting the adverse changes in spermatogenesis caused by obesity.]]> 2023/7/19 12:03:28 XU Rui,DONG Jia,CAO You-Xiang,HU Hui,XIA Qiu,WANG Peng-Yin,YU Li,FENG Wan-Yu and SHENG Lei 171true Objective To investigate the effects of Polygonatum odoratum (Mill.) Druce extract (POD) on ovarian function and inflammation of rats with polycystic ovary syndrome (PCOS).Methods Three-week-old female SD rats were used in this study. PCOS model was established by gavage of letrozole for 21 d. Then PCOS model rats were treated with different concentrations of POD by gavage. The changes of water and food intake, body mass and estrous cycle were recorded, blood related indicators were detected, serum testosterone level was measured, glucose tolerance was detected, and the morphological changes of ovarian tissue were observed by HE staining. Western blot (WB), fluorescent quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR) and immunohistochemistry were used to detect the expression of anti-Mullerian hormone (AMH), inflammatory cytokines interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) in ovarian tissues.Results Compared with the control group, the PCOS model rats had disordered estrous cycle, polycystic ovarian changes, significantly increased serum testosterone level, up-regulated ovarian AMH expression, and impaired glucose tolerance (P<0.01). The expression of IL-1β and TNF-α was increased (P<0.01). After the PCOS model rats were treated with different concentrations of POD, the estrous cycle gradually returned to normal, the number of corpus luteum in the ovary increased, the number of cystic follicles decreased, the serum testosterone level decreased, and the impaired glucose tolerance was alleviated (P<0.01). The expression of inflammatory cytokines IL-1β and TNF-α in ovarian tissue was down-regulated.Conclusion POD can improve the ovarian function of PCOS model rats and inhibit inflammatory response, and has a certain therapeutic effect on PCOS model rats.]]> 2023/7/19 12:03:30 CHE Xuan,ZHENG Biao,CHENG Di,XIE Yuan-Jie,ZENG Zhao-Ming and MO Zhong-Cheng 170true Objective Deinococcus radiodurans (D. radiodurans) is an extremophile with strong resistance to ultraviolet (UV), ionization, desiccation and chemical reagents. However, the molecular responses of this bacterium in the early recovery stage after UV irradiation are not fully understood. The aim of this work is to reveal the transcriptomic responses of D. radiodurans at this stage.Methods In this study, the transcriptomes of D. radiodurans under normal and UV irradiation culture conditions were determined by using RNA-seq technique. To identify the key genes and their regulatory relationships among the differentially expressed genes (DEGs), functional enrichment analysis was performed. Some key DEGs were selected and validated by real-time quantitative PCR. The transcriptome data from previous studies were adopted to find DEGs common to UV irradiation, ionizing radiation and desiccation stresses. The protein-protein interaction (PPI) network of DEGs was constructed; the hub genes and major modules in the PPI network were identified; functional enrichment analysis was performed for these hubs and modules.Results The results showed that the number of up-regulated genes was more than twice that of down-regulated genes in the early recovery stage after UV irradiation, and most of them were related to stress response and DNA repair. The main repair pathways in the early stage of recovery include single-strand annealing (SSA) pathway (involving genes ddrA-D), nonhomologous end joining (NHEJ) pathway (involving genes ligB and pprA) and nucleotide excision repair (NER) pathway (involving genes uvrA-C), the first two of which are for homologous recombination (HR), while the NER pathway removes pyrimidine dimers caused by UV irradiation. By comparing the transcriptome data under UV irradiation, ionizing radiation and desiccation stresses, it was found that the common responsive DEGs mainly involve Deinococcus-specific genes and the genes related to DNA/RNA metabolism. Several important hub genes and interaction modules were identified from the PPI network of DEGs, whose functions are concentrated in double-strand break repair, DNA topological change and replication.Conclusion These results indicate that in the early recovery stage after UV irradiation, a variety of genes in D. radiodurans undergo responses at transcriptome level, several repair pathways are initiated to cope with this stress, and some repair pathways are common to other stress conditions.]]> 2023/7/19 12:03:32 ZHANG Cai-Yun,QIU Qin-Tian and MA Bin-Guang 169true Objective In this study, we propose three-dimensional electrical impedance tomography (3D-EIT) combined with two-dimensional electrical impedance tomography (2D-EIT) to study the electrical characteristics of gastric volume changes and the response to changes in the gastroesophageal liquid condition with the objective of investigating the possibility of applying EIT technique to monitor gastroesophageal reflux disease (GERD).Methods Eight subjects were asked to fast 4 h before the experiment to ensure that the gastric was in an emptying condition at the start of the experiment. The subject was asked to drink 400 ml of rehydration water in two parts, 3D-EIT was applied to detect the conductivity distribution in the 3D space of the abdominal cavity at the condition of 200 ml intake and at the condition of 400 ml intake, respectively. In order to quantify the variation of electrical characteristics in different conditions, the spatial-mean conductivity () of 3D-EIT was analysed by using a paired-samples t-test. In addition, numerical simulation tools were used to establish two sizes of gastric volume models to verify that the cause of conductivity changes in the 3D space of the subject’s abdominal cavity in the experimental results is due to the changes in gastric volume. The trends of electrical characteristics when gastric cavity was filled with different ratios of rehydration water were investigated, and 2D-EIT numerical simulation was applied to reconstruct the conductivity distribution image σ.Results The results of the paired-sample t-test for spatial-mean conductivity ) in the abdominal 3D-EIT experiment in eight subjects show that the spatial-mean conductivity increased from 200 ml=0.226 in the C200 ml case to 400 ml=0.387 in the C400 ml case (n=8, P<0.05). Thus, the of subject’s abdominal gastric region increased significantly with increasing gastric volume. The 2D-EIT numerical simulation results show that the measured voltage difference (ΔV) is decreased gradually with increasing the rehydration water ratio of the gastric lumen filling. ΔV is sensitive to the ratio of gastric lumen filling rehydration water. The average voltage difference is smaller in the big gastric lumen model with the same ratio of gastric lumen filling rehydration water compared to the small gastric lumen model. is sensitive to the size of gastric lumen model. In the small gastric lumen model, the spatial-mean conductivity is increased with the ratio of gastric lumen filled with rehydration water, from A=-0.29 for 25% filling to D=-0.41 for filling 100%; in the big gastric lumen model, the is also increased with the ratio of gastric lumen filled with rehydration water, from E=-1.85 for 25% filling to H=-2.12 for filling 100%.Conclusion The 3D-EIT satisfactorily monitors the differences in electrical characteristics of different gastric volumes by 3D-EIT images. 2D-EIT shows the same trend according to ΔV, , and in different gastric volume models. Therefore, we conclude that 3D-EIT combined with 2D-EIT technique satisfactorily monitors liquid condition changes in the different gastric volumes.]]> 2023/7/19 12:03:35 ZHANG Quan-Cheng,WANG An-Xin,SUN Bo and YAO Jia-Feng 168true Objective Coxsackie virus group B type 5 (CVB5) is one of the causative agents of hand-foot- mouth disease, which can cause clinical symptoms such as fever, rash or herpes, and neurological complications or even fatalities. The innate immune response is the first line of defense against the viral infection, and the nuclear factor-κB (NF-κB) is a master regulator in the control of immune responses. However, little research has been reported on the regulation of the NF-κB mediated signaling pathway after CVB5 infection. This study explores the regulatory mechanism of virus and the host innate immune response, providing targets for the development of drugs against CVB5 infection.Methods In this study, promoter activity, proinflammatory factor and key proteins expression were detected to investigate the regulatory mechanism of CVB5 on NF-κB signaling.Results CVB5 infection inhibited the expression of proinflammatory factors and the phosphorylated p65 protein expression. Non-structural protein (NSP) of CVB5 inhibited the expression of proinflammatory factor and important proteins, such as the phosphorylated p65 and IκBα. CVB5 3CD interacted with the host polycytosine binding protein 1 (PCBP1) was performed via the STRING 11.1 database, and the PCBP1 inhibited viral replication by promoting the phosphorylation of IκBα and p65.Conclusion These results showed that CVB5 NSP negatively regulated NF-κB signaling pathway, and the PCBP1 protein which interacted with 3CD could inhibit CVB5 replication through activate the NF-κB pathway.]]> 2023/6/20 16:50:12 ZHANG Jia-Yu,TENG Pei-Ying,Lü Wei-Min,YANG Fan and CHEN Wei 167true Objective The venom of carnivorous cone snails provides a valuable source of biologically active peptides, which are composed of a complex mixture of disulfide-rich neurotoxins, commonly known as conotoxin. In this work, a novel O2 superfamily conotoxin Tx7.29 was reported, and through functional research, it is expected to discover a new analgesic drug candidate.Methods The cDNA sequence of Tx7.29 was obtained from the venom duct cDNA library of the molluscivorous Conus textile collected from the South China Sea. The mature peptide Tx7.29 with modified amino acids and disulfide bonds was synthesized and identified by mass spectrometry. Patch clamp and animal experiments were used to determine the biological function of Tx7.29.Results The cDNA of Tx7.29 encodes a 68 amino acid residues conotoxin precursor, which consists of 19 residues in the signal peptide, 28 residues in the pro-region and 22 residues in the mature peptide. Circular dichroism (CD) spectra showed that β-turn and antiparallel sheet structures were dominant contents in Tx7.29. Patch clamp experiments on the rat DRG neurons showed that Tx7.29 could significantly inhibit calcium currents, but it had no obvious effects on the sodium and potassium currents. Tx7.29 increased the hot plate latency from 0.5 to 4 h in a dose dependent manner in the mice hot plate assay and had low toxicity to ND7/23 cells.Conclusion This novel conotoxin Tx7.29 may be a useful tool for analgesic drug development and could expand our visions of the molecular targets of O2-conotoxins.]]> 2023/6/20 16:50:14 WU Yun,YANG Man-Yi,ZHANG Wei,ZHOU Mao-Jun and CAO Kun 166true Objective Orbital bone fracture has become very common in recent years, and its related treatments and therapies aim at repairing the defects. Mineralized poly (γ-glutamic acid)/2-hydroxyethyl methacrylate/poly(ethylene glycol) (γ-PGA/HEMA/PEG) polymeric cryogel is a new type of scaffolding material with an interconnective porous structure. The study aimed to examine its efficacy in the repair of orbital bone defects.Methods The γ-PGA/HEMA/PEG polymeric cryogel was prepared by the cryogelation technique. Orbital bone defects were prepared on twenty-four New Zealand white rabbit. Three groups were made depending on the implanted materials: (1) blank control group; (2) polymeric cryogel group (Gel group); (3) mineralized polymeric cryogel group (M-gel group). Specimens were taken 8 weeks and 16 weeks after implantation for gross observation, micro-computed tomography (μ-CT) and hard tissue grinding slices and tissue sections were used to observe the osteogenesis outcome.Results Radiographic results showed that the mineralized cryogel could effectively facilitate the repair of the orbital bone defect completely, with the defective area completely replaced by bone tissue. Histological results proved that the mineralized polymeric cryogel scaffolds could increase the expression of runt-related transcription factor 2 (Runx-2), alkaline phosphatase (ALP), osteopontin (OPN), and platelet endothelial cell adhesion molecule-1 (CD31), which indicated the strengthened angiogenesis and osteogenic capability after the mineralized cryogel transplantation.Conclusion The mineralized polymeric cryogel served as a potential engineering scaffold in the repair of orbital bone defects via angiogenesis and osteogenesis.]]> 2023/6/20 16:50:17 XIONG Ke,LIU Chun-Tao,WU Zhao-Ying,ZHANG Wei and ZHANG Chao 165true Objective Biosensors with multilayer biomedia are widely applied in various fields, and quantitative characterization of biosensors is still a problem for the development of sensors. This study is to quantitatively characterize the electrical properties of multilayer biomedia.Methods Combined with conformal mapping theory, the quantitative characteristics of biosensors are explored based on electrical impedance spectroscopy for clarifying the law of influence on impedance, and this study provides a basic theory for the characterization of biosensors. The impedance (Z*) of each biomedia layer is extracted, and the simulation and calculation are executed to study the correctness.Results An experimental system has been established, results show that the impedance (Z*) of the detection area continues to rise from the frequency (f) = 0.1 MHz to f=50.0 MHz in the coating process. This trend is explained that the solution in the original detection area is covered by the coating of biological medium with different dielectric properties, resulting in a decrease in the conductivity of the detection area and an increase in the impedance. Theoretical calculation results and simulation results show a great agreement with experimental results.Conclusion This study confirms that the multilayer biosensors are able to be quantitatively characterized based on electrical impedance spectroscopy and conformal mapping, which has certain practical value for the further development of biosensors.]]> 2023/6/20 16:50:25 HE Li-Dong,LI Jian-Ping,WEI Bi-Qian,WEN Jian-Ming,LIU Hao,MA Ji-Jie,HU Yi-Li,ZHANG Yu,WAN Nen and LI Ning 164true Objective Electrical impedance tomography (EIT) was applied to study the electrical characteristics of human calf muscles under neuromuscular electrical stimulation (NMES), with the objective of using EIT as a long-term monitoring method to visualize the effectiveness of NMES training on human calf muscles.Methods Sixteen subjects were randomly assigned to a control group (CG, n=8), which kept a normal lifestyle and without NMES or other muscle training; an optimal voltage intensity training group (OG, n=8), in which 23 min of NMES training was performed on the right calf using commercial NMES equipment 3 times a week for 5 weeks. EIT was applied to obtain the conductivity distribution before the start of each training cycle on Monday. Bioelectrical impedance analysis (BIA) was also used to measure the extracellular volume ratio (ECW/TBW) βrl in the right leg.Results In the CG, subjects showed no significant differences in the electrical characteristics of right calf muscle and in the extracellular volume of right calf compared to the first week. However, in the OG, subjects showed the same tendency of increase in the spatial-mean conductivity <σ>M1 of right calf muscle M1 muscle compartment and βrl of right leg compared to the first week.Conclusion NMES training caused an increase in the stimulated muscle compartment spatial-mean conductivity <σ>M1, with the most dramatic increase in <σw2>M1 of second week, followed by an increase in the third, fourth and fifth weeks compared to the previous week with less growth rate. This implies that a significant increase in muscle fiber volume and sarcoplasmic hypertrophy occurred in the second week, after which the subjects gradually adapted to NMES training and the physiological response of the muscle slowed but continued to grow.]]> 2023/6/20 16:50:28 SUN Bo,Darma Panji Nursetia,ZHANG Quan-Cheng and YAO Jia-Feng 163true Objective Giving up arises when an animal repeats a behavior to avoid danger or to obtain reward without success. It is a common and fundamental behavior that has been extensively studied in model animals such as mice, but the underlying neural mechanisms have not yet been well elucidated. Zebrafish larvae are important model organisms in neuroscience due to their suitability for whole brain imaging. Negative stimuli such as continuous electric shock have been used to induce giving up behavior in zebrafish, however, whether reward stimuli can induce giving up in zebrafish has not been reported.Methods By presenting larval zebrafish visual stimuli of paramecium, the hunting behavior of zebrafish on virtual food was examined, and the changes of the zebrafish predation frequency and predation duration over time were compared.Results Virtual visual stimulation of food can induce the predatory behavior of zebrafish. After receiving virtual stimuli for 25 min, the predation frequency and single predation duration of over 8 days post fertilization (dpf) zebrafish decreased significantly.Conclusion Our study enriches the paradigm of zebrafish give up behavior. The experimental results show that virtual food stimuli lacking real rewards can induce zebrafish to abandon predation. The current behavioral paradigm provides a new method to study animal give up behavior and the underlying neural mechanisms.]]> 2023/6/20 16:50:41 ZHANG Xiang-Yu,CHAI Yu-Ming,QI Ke-Xin and WEN Quan 162true Objective Understanding the dynamics of protein adsorption at the aqueous gold interface is key to advancing the development of many applications based on gold nanostructures such as plasmonic sensors and biomedical materials. However, the protein conformation in metallic nano-slits has not been fully studied. In this paper, the spacing effect of protein conformation changes in gold nano-slits is studied by building gold atomic layers with different nano-slits using molecular dynamics simulations.Methods We used GOIP-CHARMM force fields with GROMACS to simulate the dynamics of Aβ1-42 protein monomers between Au(111) gold atomic interfaces to study the dynamics of Aβ protein and conformational changes in aqueous environments, 5.0 nm nano-slit, 5.5 nm nano-slit and 8.5 nm nano-slit.Results When the nano-slit spacing is increased from 5.0 nm to 8.5 nm, the interaction of Aβ protein with the gold interface is varied from double-interface adsorption, single-side interface adsorption to no adsorption.Conclusion The interaction of Aβ protein with the metallic interface is greatly affected by the spacing of gold nano-slits. As the slit spacing and the distance of the protein from the interface changes, the protein could exhibit 3 distinct states: single-side surface adsorption, two interface adsorption and no adsorption. We anticipate that the dynamic features of protein adsorption to Au(111) interfaces of nano-slits showed in this work can be extended to other nanostructures and proteins. The results could find applications in design of biosensors for diagnosis of Alzheimer’s disease.]]> 2023/6/20 0:00:00 WANG Lei,WANG Yan-Hong,WU Jing-Zhi and LI Meng-Wei 161true Objective Deep brain stimulation (DBS) utilizes sustained high-frequency stimulation (HFS) of electrical pulses to modulate neuronal activity. The therapy is expected to be used to treat more brain disorders. To deeply understand the mechanisms of the HFS to advance the DBS development, the present study investigates the effect of axonal HFS on neuronal somata during HFS-induced axonal block.Methods Antidromic high-frequency stimulation (A-HFS) with a 100-Hz pulse frequency and a 1-min duration was applied at the axons of pyramidal neurons in the hippocampal CA1 region of anesthetized rats. To investigate the responses of somata, a multi-channel microelectrode array with a vertical linear configuration was implanted to record the evoked potentials in the lamellas around the somata of CA1 pyramidal neurons at the upstream area of stimulation site, including the antidromic population spikes (APS) evoked by the pulses of A-HFS as well as the orthodromic population spikes (OPS) evoked by orthodromic test pulses applied during the A-HFS. Current-source densities (CSD) of the evoked potentials were calculated to evaluate the generation and propagation of action potentials around the somata of pyramidal neurons during A-HFS.Results A-HFS on the axons of pyramidal neurons slowed down the propagation speed of both antidromic and orthodromic excitations around somata. In addition, the occurrence and recovery of the changes of somata were slower than the A-HFS-induced axonal block.Conclusion Axonal HFS can induce soma alterations that might be caused by changes in membrane potentials nearby somata. The finding is helpful for deeply revealing the mechanisms of electrical stimulations of brain nervous system.]]> 2023/3/22 0:00:00 XU Yi-Peng,FENG Zhou-Yan,YUAN Yue,HU Yi-Fan,YE Xiang-Yu and WANG Zhao-Xiang 160true Objective To investigate the role of histone deacetylase 3 (HDAC3) in the differentiation and function of peripheral CD4+ T cells.Methods CD4cre enzyme mediated HDAC3 heterozygous gene deletion mice (Hdac3fl/flCD4cre+/-) and wild-type normal control (Hdac3fl/fl, WT) mice were used. The effects of HDAC3 deletion on the proportion and number of peripheral CD4+ and CD8+ T cells were detected by flow cytometry. The effects of HDAC3 deletion on the expression of IFN-γ, IL-4 and IL-17A in CD4+ T cells and Tfh cells were detected under the in vitro PMA and Ionomycin stimulation. The effects of HDAC3 deletion on the expression of IFN-γ, IL-4 and IL-17 in serum were detected by ELISA. The naive CD4+ T cells of Hdac3fl/flCD4cre+/- and WT mice were sorted and cultured in Th1 and Th2 differentiation conditions respectively. The effects of HDAC3 deletion on the expression of Th1, Th2 and Th17 related cytokines and their specific transcription factors were detected by intracellular staining. The effects of HDAC3 deletion on the expression of genes related to CD4+ T cell differentiation subsets were detected by gene expression microarray. The mice treated with streptozotocin (STZ) were used to construct type 1 diabetes mellitus (T1DM) disease model, and the effects of HDAC3 deletion on the pathogenesis of T1DM were detected.Results Compared with WT mice, the proportion and number of peripheral CD4+ and CD8+ T cells in Hdac3fl/flCD4cre+/- mice decreased significantly. The expression of IFN-γ in CD4+ T cells and serum of Hdac3fl/flCD4cre+/- mice decreased significantly, while the expression of IL-4 and IL-17A increased significantly, and the proportion of Tfh cells also increased significantly. HDAC3 deletion inhibited the differentiation of CD4+ T cells into Th1 cells, but promoted their differentiation intoTh2 cells. Microarray analysis showed that the deletion of HDAC3 resulted in the decrease of gene expression in Th1 cell lineage, while the increase of gene expression in Th2, Th17 and Tfh cell lineage. Under the condition of STZ induction, HDAC3 deletion inhibited the development of T1DM and the differentiation of CD4+ T cells into Th1.Conclusion HDAC3 promotes the differentiation of peripheral CD4+ T cells into Th1 cells and aggravates the occurrence of T1DM.]]> 2023/3/22 0:00:00 GUO Han,LI Wen-Ting,ZHANG Ting,ZHANG Ai-Hong,ZHENG Ai-Hua,TIAN Feng and ZHENG Quan-Hui 159true Objective The efficacy of repeated transcranial magnetic stimulation (rTMS) as a noninvasive neuromodulation technique in patients with Parkinson’s disease (PD) has not been fully Validated. In this study, we combined clinical scale evaluation, brain electrical source and brain network to explore the effects of high-frequency repetitive transcranial magnetic stimulation on patients with akinetic-rigid Parkinson’s disease (AR-PD).Methods A total of 18 patients with AR-PD were included. The EEG signals were traced by standard low-resolution electromagnetic tomography (sLORETA), then the brain functional network was constructed by complex network theory, and the network topological characteristics were compared and analyzed from the perspective of collaborative work between brain regions.Results The results showed that there were significant differences in prefrontal cortex and primary motor cortex after magnetic stimulation (P<0.05). The network connectivity of brain regions associated with motor sensory production, motor planning, and motor execution was significantly enhanced (P<0.05) and the change in the mean cluster coefficient of topological features of brain functional networks was significantly correlated with the change in Parkinson’s uniform rating scale score (P<0.05).Conclusion It is speculated that rTMS improves the information transmission ability of AR-PD patients from motor sensation generation to motor execution. This study can provide a theoretical basis for the relationship between the improvement of motor symptoms of AR-PD patients by rTMS and the reintegration of sensorimotor network.]]> 2023/3/22 0:00:00 LI Jia-Li,YIN Ning,YAO Yao,FENG Ke-Ke,LI Run-Ze,LIU Shuo,YIN Shao-Ya and XU Gui-Zhi 158true Objective Locomotor training has been shown to preferentially affect muscle function in many chronic diseases which has been used in the treatment of sarcopenia. In this study, we propose electrical impedance tomography (EIT) to study the electrical characteristics of physiological response in calf muscle compartments induced by locomotor training, which is intended to visualize the effectiveness of locomotor training on the increase in volume of calf muscle fibers.Methods Experimental subjects were asked to perform unilateral heel rises on the right and left legs on 5 consecutive experimental days. EIT was applied to detect the conductivity distribution of calf muscles after locomotor training. In order to quantify the effectiveness of locomotor training, a paired sample t-test was used to analyze the spatial-mean conductivity <σ> from reconstructed EIT images.Results The results show that the spatial-mean conductivity <σ>M1 of M1 compartment (which is recognized as the position of the gastrocnemius muscle) is significantly increased in post-training by locomotor training. In addition, the spatial-mean conductivity in pre-training <σpre>M1 shows an increasing tendency in EIT measurement results on 5 consecutive experimental days. The lean mass of all subjects in pre- and post-training parts which was performed in the morning of experimental day 1 is linearly related to <σ>M1; the increase in <σ>M1 and spatial-mean conductivity difference ratio Δ<σ>M1 between pre- and post-training followed the same tendency as extracellular water/intracellular water (ECW/ICW) τ by ECW/ICW difference ratio Δτ.Conclusion Since conductivity is sensitive to changes in extracellular volume which correlates with sarcoplasmic hypertrophy. The <σpre>M1 tended to increase during 5 consecutive experimental days of EIT measurements, which implies an increase in muscle fiber volume with sarcoplasmic hypertrophy occurring. Therefore, EIT is able to effectively detect the effectiveness of locomotor training to increase the volume of human calf muscle fibers.]]> 2023/3/22 0:00:00 SUN Bo,Sejati Prima Asmara and YAO Jia-Feng 157true Objective Wool is a high-grade raw textile material, and the physical properties of wool are directly related to the quality of the wool. The present study was to search for the genes affecting wool traits and to explore the complex molecular mechanism affecting wool traits.Methods This study selected 3 Suffolk sheep and 3 small-tailed Han sheep and took samples of their back skin tissue using RNA sequencing (RNA-seq) and proteome sequencing analysis of genes, proteins, and related signalling pathways that cause differences in wool traits.Results RNA-seq showed that after sequencing, 230 406 674 raw data points and 222 049 370 clean data points were obtained, of which the percentage of Q20 bases was over 99.9%, and the percentage of Q30 bases was over 98%. With a fold change (FC)≥1.4 or FC≤0.714 and P<0.05 as the standard, 1 213 differentially expressed genes (DEGs) were screened out, among which there were 644 upregulated genes and 569 downregulated genes in Suffolk sheep in comparison with small-tailed Han sheep. The gene ontology (GO) enrichment found that intermediate filament, calcium ion binding, and keratin filament were significantly enriched, indicating that they might be related to wool traits. The Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment found that the signalling pathway affecting wool traits might be the ECM-receptor interaction. Proteome sequencing showed that with FC≥1.4 or FC≤0.714 and P<0.05 as the standard, 99 differentially expressed proteins (DEPs) were screened out, among which there were 47 upregulated proteins and 52 downregulated proteins in Suffolk sheep in comparison with small-tailed Han sheep. The GO enrichment found that intermediate filament was significantly enriched, indicating that it might be related to wool traits. The KEGG enrichment found that the signalling pathways affecting wool traits might be the peroxisome proliferator-activated receptor (PPAR) and the ECM-receptor interaction. A combined analysis of RNA-seq and proteome sequencing found that a total of 15 significantly different genes were detected in both RNA-seq and proteome sequencing, of which 13 were positively correlated and 2 were negatively correlated. Intermediate filament was significantly enriched, KRT35, KRT13 and KAP13-1-like genes might be the key candidate genes to affect wool traits. The PPAR signalling pathway was significantly enriched and might be a key candidate pathway to affect wool traits. The FABP4 gene might be a key candidate gene to affect wool traits.Conclusion Among them, KRT35 might affect wool bending and diameter, KRT13 might affect wool differentiation, while KAP13-1-like might affect wool hardness and toughness, FABP4 might affect wool diameter. These results will expand our understanding of the complex molecular mechanisms affecting sheep wool traits and provide a basis for subsequent studies.]]> 2023/3/22 0:00:00 WU Jin-Qiang,HAO Xiao-Jing,ZHAO Hong-Xia,DONG Ya-Jie,WANG Rong,ZHANG Peng-Xiang,WANG Hai-Dong and HE Xiao-Yan 156true Objective In multicellular organisms, cell communication allows multiple cells to coordinate with each other and involved in various important biological processes, and the abnormal cell communication pattern have also been demonstrated to be associated with many diseases. In the past decades, the rapid development of single-cell RNA sequencing technology makes it possible to study cell communication pattern through constructing cell communication network based on inferring ligand-receptor interactions, and many methods have been proposed. However, most of them only consider the expression of ligands and receptors to quantify the cell-cell interaction strength, but ignore the effect of specific receptor to its downstream gene regulatory network which reflects whether that receptor participates in particular biological process or not. To fill this gap, we propose a novel method (named IRRG) to construct cell communication networks at cell type level.Methods Our IRRG algorithm consists of the following three main steps. (1) IRRG uses the signaling pathway database to construct the gene regulatory network for each receptor in each cell type, and then calculates receptor impact score (RIS) for each receptor based on a random walk algorithm. RIS represents the degree of receptor impacts on downstream genes during cell communication. (2) IRRG uses permutation test to identify ligand-receptor pairs with significant specificity across cell types in order to discover more biologically significant cellular communication phenomena. (3) IRRG combined ligand-receptor co-expression with RIS to calculate the strength of all ligand-receptor interactions between cell types, and then constructed cellular communication networks.Results To validate the effectiveness of our IRRG, we construct cell communication networks of mouse epidermal tissue and human kidney cancer tissue separately. For mouse interfollicular epidermis (IFE) dataset, IRRG constructs biologically meaningful cellular communication networks, reasonably infers the different roles played by cells at different levels of epidermal tissue in cooperation, and discovers ligand-receptor pairs that are closely related to epidermal physiological processes. We also verified the robustness of the IRRG calculated receptor impact score, indicating that the IRRG calculated receptor impact score can reasonably reflect the signaling of cellular communication processes. By counting the supporting literature possessed of the top ranked ligand-receptor pairs, we also indirectly verified that the cell communication network constructed by IRRG is more reliable. In addition, we identify the cellular communication patterns of tumor tissues in clear cell renal cells (ccRCC) using IRRG and analyze how tumor cells influence normal cells to complete their own growth and development or migration process through cellular communication, further understanding the tumor microenvironment.Conclusion In this paper, we propose a novel method for constructing cellular communication networks named IRRG which integrating receptor-regulated gene expression information through random walk. Case studies on two datasets show that IRRG can construct biologically significant cellular communication networks, which can help us understand the mechanism of biological processes from the perspective of cellular communication. The source code and associated datasets used in this work are publicly available at https://github.com/NWPU-903PR/IRRG.]]> 2023/3/22 0:00:00 GUO Shu-Qi,ZHANG Shao-Wu,LI Yan and ZHANG Shi-Hua 155true Objective Anticancer drug combination therapies are a promising therapeutic strategy. Drug combinations exhibiting a highly synergistic effect are crucial to improve the treatment for specific cancers. However, identifying such combinations is very complicated and difficult due to the tremendous screening cost. The availability of large-scale high-throughput combination screening data provides opportunities for computational approaches. The purpose of this study is to optimize the high-throughput virtual screening of anticancer drug combinations in a completely data-driven and computational modeling way, and provide theoretical reference for “old drugs repositioning as new combinations”.Methods Inspired by the matrix completion, we present a nuclear norm regularization-based model, termed NNRM, to predict synergy scores and synergy status of anticancer drug combinations. Symmetric observation matrixes of synergy scores were constructed for given cell lines; a folding technique was employed to sparse the observation matrix; alternating direction multiplier method and soft threshold estimation were applied to solve the model.Results NNRM achieved expected predictive result on the dataset released by O’Neil’s team, the root mean square error of the synergy score prediction was 14.78, and the accuracy of the synergy status prediction was 0.94. It is not only significantly superior to the Random Forest and the Support Vector Machine, but also completely comparable to the state-of-the-art deep learning models including DeepSynergy, Deep learning+PCA and AuDNNsynergy. Moreover, NNRM effectively filled the missing synergy scores most of which are consistent with existing research or clinical practice.Conclusion NNRM could predict the synergistic effect of large-scale drug combinations in batches, which greatly lowers the data requirements by existing models, reduces the computational cost, and shortens the screening time. It indicates that NNRM is an alternative tool for high-throughput virtual screening of anticancer drug combinations.]]> 2023/3/22 0:00:00 SHI Lei-Jing,WANG Bo,ZHANG Shan,REN Fu-Quan and Li Yu-Shuang 154true Objective The comparative proteomic study on the paired lesional epidermis (LE) and non-lesional epidermis (NLE) from vitiligo patients to identify the differentially expressed proteins (DEPs) between LE and NLE, and to further explore the molecular mechanism of pathogenesis of vitiligo.Methods Firstly, the in solution digestion condition for proteins from epidermis were optimized to sequential tandem digestion with Lys-C and trypsin. Secondly, tandem mass tag (TMT) based quantitative proteomic strategy was performed to compare the proteome profile of the paired LE and NLE from three stable non-segmental vitiligo subjects, and differential expressed proteins (DEPs) were identified. At last, the functional enrichment analysis was performed via bioinformatics tool and database (GO, KEGG, STRING, GSEA).Results The optimal sequential tandem digestion condition was the combination of Lys-C (enzyme∶substrate, 1∶100) and trypsin (enzyme∶substrate, 1∶50). A total of 4 496 proteins were identified, and of which 181 were DEPs between LE and NLE from vitiligo patients. Bioinformatics analysis showed that DEPs were mainly related with metabolism, immunity, redox and cell adhesion. Among them, the 119 up-regulated proteins are mainly involved in the processes of keratinization, transcription, oxidative stress, and proteolysis. The 62 down-regulated proteins are mainly involved in intracellular transport, glutathione metabolism and actin filament capping.Conclusion The comparative proteomic study revealed that there were functional differences in keratinization, immunity, lipid metabolism and redox between LE and NLE in vitiligo patients. PRDX1, PRDX2, EEF2, ITGB1, SPTBN2, ANXA1 and PFKL were found as the key proteins to disfunction of LE.]]> 2023/2/22 0:00:00 Tuerxun Ailikemu,CHEN Xiu-Lan,Talifu Ainiwaer,LI Na,WANG Ji-Feng,CAI Tan-Xi,GUO Xiao-Jing,DING Xiang,XIE Zhen-Sheng,NIU Li-Li,ZHANG Meng-Meng,Abbas Ghulam,Aisa Haji Akber and YANG Fu-Quan 153true Objective This study was designed to observe different durations of simulated weightlessness induced depressive-like behaviors and the change of superoxide dismutase (SOD) and catalase (CAT) in hippocampi of rats, so as to explore its effects and possible mechanism.Methods Weightlessness was simulated by hindlimb suspension by tails (HLS) in rats. The rats were divided into the control and the HLS groups for each respective week (s), i.e., 1, 2, 3, and 4. The depressive-like behaviors in rats were investigated by using the open field test (OFT), novel object recognition test (NORT), forced swim test (FST), followed by hippocampal protein level estimation of SOD and CAT by ELISA.Results The OFT results showed increased stagnation time among HLS rats in each respective group compared to their corresponding control groups (P<0.05, P<0.01). In the NORT, the latency of exploration increased, while the number and the time of exploration decreased in the HLS groups in each week, compared to their corresponding control rats (P<0.05, P<0.01). Similarly, FST displayed enhanced immobility with the reduced climbing rate in HLS rats in each respective week, compared to the normal rats (P<0.05, P<0.01). Simulated weightlessness significantly reduced the SOD and CAT levels in the hippocampi in HLS rats compared to the normal rats (P<0.05, P<0.01).Conclusion Results suggest that short or long weightlessness could induce depressive-like behaviors in rats.]]> 2023/2/22 0:00:00 ZHANG Mi-Jia,WANG Meng-Di,AYAZ Ahmed,DONG Li,YAO Qin-Wei,LI Ying-Hui,QU Li-Na,CHEN Shan-Guang,LIU Xin-Min and WANG Qiong 152true Objective Since the first discovery that CRISPR-Cas system provides adaptive immunity of prokaryotic hosts to virus and other mobile genetic elements (MGEs). Numerous studies yielded vital insights into the immune mechanisms, and CRISPR-Cas system has been wildly utilized in gene editing and related research efforts. In the three major immune stages—adaptation, expression and maturation, interference spacer sequences play important roles separately. Although PAM (protospacer adjacent motif) determines the identification of targeted genes by CRISPR-Cas system, what drives the selection of specific genes and reservation on the CRISPR array remains uncertain. To explore the targeting characteristics of CRISPR-Cas systems, the virome of Yangshan harbor surface water and the CRISPR-Cas spacers available in public datasets were subjected to analysis.Methods Based on BLAST searching, viral sequence identification, gene function prediction, and gene conserved domain annotation, the final analysis results were obtained.Results As a result, 25 391 double-stranded DNA viral sequences were identified in the virome; 265 open reading frames (ORFs) were predicted for 238 sequences, and 134 CRISPR-Cas spacer sequences yielded 315 viral hits. 128 viral ORFs and 135 hits were functionally annotated, and the top 5 hits including terminase, capsid protein, portal protein, peptidase, and DNA methyltransferase. The matching of spacer (host)-protospacer (virus) often occurred in conserved domains or key structural domains of viral functional genes. Meanwhile, the number of CRISPR-Cas spacer sequence matches for Type_I systems under the Class 1 was much higher than that for other types of systems, accounting for 89.0% of the total. The results show that the CRISPR system will specifically identify and act on key functional genes of the virus.Conclusion The results of this study reveal the targeting specificity of the CRISPR-Cas system, showing new insights and providing new evidence for a better understanding of the mechanisms of virus-host immune interactions.]]> 2023/2/22 0:00:00 LI Cong,TIAN Jin-Zi and WANG Yong-Jie 151true Objective DUF538 (domain of unknown function 538) domain containing proteins are known as putative hypothetical proteins in plants. Until yet, there is no much information regarding their structure and function.Methods In the present research work, the homologous structures and binding potentials were identified between plant/mammalian lipocalins and plant DUF538 protein by using bioinformatics and experimental tools including molecular dynamics simulation, molecular docking and recombinant technology-based techniques.Results Molecular docking analysis of their interactions with lipidic ligands including cholesterol and palmitic acid revealed the similar and comparable binding potentials between DUF538 and lipocalin proteins. Both the test proteins were found to have more affinity to cholesterol molecule in compare to palmitic acid. By using recombinant technology-based experiments, the heterologously expressed and purified fused product of DUF538 protein exhibited about 61% cholesterol binding ability.Conclusion As a conclusion, plants DUF538 protein family was predicted to be the structural and may be the functional homologues of plants/animals lipocalin superfamily.]]> 2023/2/22 0:00:00 GHOLIZADEH Ashraf 150true Objective The reservoir of human immunodeficiency virus (HIV) latently infected cells is the major obstacle for eradication of acquired immunodeficiency syndrome (AIDS). Due to the noisy environment and multiple influencing factors in the organism, current dynamical models cannot reach a common understanding of the molecular mechanism of HIV latency. In this work, through a new dynamical structure decomposition, the deterministic part of the equation can be separated from the stochastic noise. Thus, the fixed-point analysis of ordinary differential equation is enough to obtain the different steady states of the system.Methods We established a dynamical model of HIV transcription process by using continuous stochastic differential equations, which simplifies the dimensions of equations needed to describe the system and increases the explorable space of the model. Different states between latency and activation of virus and their relations can be intuitively represented by potential functions and probability distribution functions.Results Based on our model, the influence of different dynamical parameters on stability is quantitatively analyzed, the parameter ranges of the system in bistable and monostable states are obtained respectively. The theoretical basis of this work is verified by comparing the effects of different factors on dynamic bifurcation with the results of biological experiments.Conclusion This paper goes beyond previous discrete stochastic methods, and can quantitatively analyze the dynamic mechanism of HIV transcriptional regulation through ordinary differential equations, which is beneficial to the promotion to deal with the high-dimensional situation, and further study the occurrence and development of AIDS in vivo, so as to guide the design of experiments and search for clinical treatment.]]> 2023/2/22 0:00:00 XIONG Rui-Qi,SU Yang and AO Ping 149true Objective Repetitive transcranial magnetic stimulation (rTMS) has been widely used in clinics to realize the treatment of neurological and psychiatric diseases, especially in depression and pain. In recent years, many studies have applied it as an adjuvant approach to treatment of Parkinson’s disease (PD) in order to relieve the patient’s motor symptoms and improve motor function. At present, the unified Parkinson’s disease scale and motor task are used to evaluate the adjuvant treatment effect of rTMS on motor symptoms, but there are few studies exploring the regulatory mechanism of high-frequency rTMS stimulation.Methods In this study, 10 Hz rTMS over the primary motor cortex (M1) on the contralateral side of the limb symptom onset side in 16 PD patients. By comparing the changes of brain neuron activity and the interactions between brain function before and after stimulation, we investigated the regulatory effect of repeated sessions of M1 high-frequency rTMS on brain neuron activity in PD patients.Results The results showed that after 10 d of M1 stimulation, beta oscillations increased and gamma oscillation significantly decreased (P<0.05) in motor cortex, and the connectivity between the frontal and parietal cortex was reduced (P<0.05).Conclusion It shows that 10 Hz rTMS mainly changes the beta and gamma rhythm in PD patients. The changes of beta and gamma oscillations in motor cortex may be related to the improvement of motor function, and the changes in prefrontal gamma oscillations may be due to inhibits abnormal neuron firing activities and modulates the patient’s motor control function.]]> 2023/1/16 15:04:20 LI Run-Ze,YAO Yao,FENG Ke-Ke,YANG Shuo,LI Jia-Li,CHENG Yi-Feng,YIN Shao-Ya and XU Gui-Zhi 148true Objective Beta-secretase 1 (BACE1) is the key enzyme for amyloid β (Aβ) production in the Alzheimer’s disease (AD) brain. The dystroglycan (DG) protein anchors astroglial endfeet onto cerebral blood vessels forming glia limitans, a supportive element in the blood-brain barrier. An untargeted proteomics study predicted that BACE1 downregulates DG expression. Here, this study investigated whether BACE1 modulates the protein levels of DG and its hypothetic mechanism.Methods Transient transfection technique was used to express target protein in HEK-293T cells and in primary mouse astrocytes. And the protein levels of targets were analyzed by Western blot. Quantitative polymerase chain reaction and co-immunoprecipitation were used to explore the potential mechanisms of BACE1-dependent regulation of DG.Results This study found that addition of BACE1 resulted in significantly lower levels of β subunit DG (β-DG) protein, both in HEK-293T cells and in primary mouse astrocytes. And in HEK-293T cells, this down-regulation of β-DG protein dependent on the enzyme activity of BACE1.Conclusion BACE1 lowers β-DG protein levels in HEK-293T cells and in mouse astrocytes.]]> 2023/1/16 15:04:23 YUAN Fang,WU Zu-Jun,LONG Zhen-Yu,BI Dan-Lei and SHEN Yong 147true Objective The aim of this study is to investigate the effect of matrix stiffness changes on the differentiation of neural stem cells (NSCs) and the underlying mechanism.Methods A rat model of spinal cord injury was constructed. Polyacrylamide gel substrates with different stiffness (0.7 kPa, 40 kPa) were prepared, and were cultured with primary rat NSCs. NSCs cells were transfected with piezo type mechanosensitive ion channel component 1 (Piezo1) shRNA plasmid. Immunofluorescence staining was used to detect the percentage of positive cells for the neuron marker doublecortion(DCX) and the astrocyte marker GFAP. Immunohistochemistry experiment and Western blot were used to detect the protein expression of Piezo1.Results The results showed that compared with the 0.7 kPa group, the number of DCX-positive cells in the 40 kPa group increased, while the number of GFAP-positive cells decreased, as well as the protein expression of Piezo1 increased. The expression of Piezo1 was significantly up-regulated in the injured tissue of rats with spinal cord injury compared with the sham group. Silencing of Piezo1 reverse the effect of 40 kPa matrix stiffness on NSCs proliferation, as indicated by decreased number of DCX-positive cells, and increased number of GFAP-positive cells increased. Further studies found that knockdown of Piezo1 leads to decreased expression of collagen IV and fibronectin. Recombinant fibronectin reversed the effect of sh-Piezo1 on the differentiation of NSCs.Conclusion Rigid base stiffness regulates the differentiation of NSCs by promoting the expression of Piezo1 protein and up-regulating the expression of type IV collagen and fibronectin. This study provides a new perspective for the treatment of spinal cord injury based on biomaterials.]]> 2023/1/16 15:04:34 XU Wang-Yang,HUANG Li-Shan,YAO Shun,HUANG Zi-Xiang,ZHANG Li,ZHANG Hui and WANG Ye-Yang 146true Objective To study the anti-breast cancer effects and molecular mechanisms of syringin, and to provide a theoretical basis for the clinical application of syringin.Methods The inhibitory effect of syringin on the proliferation of breast cancer cells was measured with MTT assay. Trypan blue, TdT-mediated dUTP nick-end labeling (TUNEL), and Annexin V-FITC/PI staining were used to detect apoptosis. Caspase-3 activation was detected via Western blot to determine whether apoptosis occurred. The expression of apoptosis-associated protein B-cell lymphoma-2 (Bcl-2) was detected and the effect of syringin on the mitochondrial apoptosis pathway was investigated via JC-1 staining. The PI3K agonist Recilisib was used for comparison. qRT-PCR and Western blot were used to assess the role of syringin in regulating the PI3K/Akt/mTOR pathway and inducing the apoptosis of cancer cells.Results Syringin had a time- and dose-dependent inhibitory effect on the proliferation of breast cancer cells and induced their apoptosis. A further study showed that after syringin treatment, Caspase-3 was activated, Bcl-2 expression decreased, the mitochondrial membrane potential was significantly reduced, and the mRNA and protein expressions of PI3K, Akt, and mTOR were not significantly changed, but the protein phosphorylation levels were significantly decreased. Recilisib partially limits the effect of syringin on the apoptosis of breast cancer cells.Conclusion Syringin has a good inhibitory effect on MDA-MB-231 and MCF-7 breast cancer cells. It can inhibit cell proliferation and induce mitochondrial apoptosis by inhibiting the activation of the PI3K/Akt/mTOR signaling pathway. Syringin is a potential anti-breast cancer drug.]]> 2023/12/22 11:25:40 SHI Ya-Qian,LI Xin,HUANG Si-Yuan,OU Ming-Kun,LI Hong-Na,LU Min,GENG Meng-Li and OU Ye-Tao 145true Objective In recent years, accumulating evidence indicated that epigenetic alterations may be used as the potential biomarkers to detect toxicity caused by metal nanoparticles (MNPs). Thus, the effects of MNPs exposure on the phosphorylation of histone H3 at serine 10 (p-H3S10) were clarified, and the changes of whole gene expression in cells exposed to typical MNPs were explored in this study to provide a theoretical basis for screening the biomarkers of early toxicity of MNPs.Methods The effects of ten kinds of MNPs exposure on the p-H3S10 were evaluated using Western blotting and flow cytometry. In addition, the effects of copper oxide (CuO) NPs, which is a typical MNPs, on the whole gene expression of cells were explored at the transcriptional level by RNA sequencing technology.Results All the tested MNPs except nickel oxide NPs induced p-H3S10 to different degrees. Among, upregulation of p-H3S10 was observed at immediately after CuO-NPs and zinc oxide NPs treatment, and continued for 10 h. Moreover, p-H3S10 increased slightly after aluminium oxide NPs and antimony (III) oxide NPs treatment, and reached the highest point at 0.5 h. After that, p-H3S10 began to decrease and obtained the lowest value at 2 h. However, p-H3S10 began to rise again after treatment over 2 h until the final observation time point. In addition, p-H3S10 was temporarily induced after treatment with iron (II,III) oxide NPs, silica NPs, cobalt (II,III) oxide NPs, chromium (III) oxide NPs or titanium dioxide NPs, but rapidly ceased. Further analysis indicated that MNP-induced p-H3S10 was highly related to the cellular uptake of MNPs, and the sustained release of ions from MNPs inside cells might generate p-H3S10 for an extended period after the initial uptake of MNPs. In addition, RNA sequencing analysis revealed that CuO-NPs treatment caused significant differential expression of 275 genes (P<0.05), 185 of which were upregulated and 90 of which were downregulated. Gene Ontology (GO) analysis showed that the majority of differentially expressed genes (DEGs) were involved in regulation of signaling, transcription factor activity and kinase activity. Through analyzing enriched GO pathways related to stress, two prominent membrane-activated cascades emerged: the mitogen-activated protein kinase (MAPK) cascade, and Janus kinase/signal transducers and activators of transcription (JAK/STAT) cascade. Genetic markers specific to extracellular signal-regulated kinases ERK1/ERK2 regulation, stress-activated MAPK cascades, and JAK/STAT cascade were also significantly altered after CuO-NPs treatment. It was observed that a large number of genes related to MAPK tyrosine/serine/threonine phosphatase activity were differentially expressed due to CuO-NPs treatment. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis showed strong associations between the DEGs and signal transduction, signaling molecules and interactions, immune and endocrine systems. Moreover, several DEGs were also correlated with transport, catabolism, cell growth and cell death. MAPK cascades were also significantly upregulated after CuO-NPs exposure.Conclusion These results indicated that early induction of p-H3S10 by MNPs is highly related to the cellular uptake of MNPs, and the persistent release of ions from MNPs inside cells might generate p-H3S10 for a long time after the initial uptake of MNPs. Taken together, the p-H3S10 has potential as a suitable candidate biomarker for evaluating the toxicity of MNPs.]]> 2023/12/22 11:26:09 ZHAO Xiao-Xu,ZHOU Lu-Lu,HUANG Yu-Chun,XIONG Yi-Fan,LIN Shou-Kai and LI Yu-Shuang 144true Objective To construct an analysis process for identity-by-descent (IBD) algorithm to predict distant relatives and evaluate the prediction accuracy.Methods 253 family samples were detected by using high-density whole genome single nucleotide polymorphism (SNP) chip. IBD algorithm was used to predict the genetic relationship between pairs of individuals, and the prediction accuracy was evaluated. The number of SNPs was randomly reduced to evaluate the effect of SNP numbers on the accuracy of the algorithm prediction.Results Among 1-7th kinship degree, the average confidence interval accuracy of IBD algorithm was 94.72%, and the prediction credibility was 99.77%. The false negative of IBD algorithm was found when kinship degree is 6 or higher. When the number of SNPs decreases, the prediction accuracy will decline to a certain extent.Conclusion The IBD algorithm can accurately predict the genetic relationship within the seventh kinship degree, and it has important application value in population genetics, forensic genealogy inference and other fields.]]> 2023/12/22 11:27:00 LIU Jing,LI Jing,YANG Lan,GUAN Shan-Shan,WEI Yi-Liang,ZHAO Wen-Ting,JIANG Li,ZHAO Dong and LI Cai-Xia 143true Objective The warm sensors located in keratinocytes have the ability to directly detect warm temperature, with TRPA1 in dorsal root ganglion (DRG) neurons being a potential mediator of the downstream transduction process. The present study aimed to elucidate the mechanism by which warm information is transmitted from keratinocytes to TRPA1.Methods TRPA1-transfected HKE293T cells as well as DRG neuron cells from wild-type and TRPA1 knockout mice were cultured separately, and stimulated with warm temperatures using a perfusion apparatus. Single cell calcium imaging was used to monitor calcium influx during stimulation, and the role of H2O2 in this process was also examined. Additionally, RNA-seq analysis was performed on primary keratinocytes cultured at different temperatures to identify potential candidates responsible for keratinocytes-TRPA1 warm transduction.Results Our finding indicated that TRPA1-transfected HEK293T cells or DRG neurons could be activated by warm temperature in the presence of H2O2. However, when TRPA1 was knocked out or blocked by HC-030031, the H2O2-potentiated warm response was significantly reduced. Moreover, Chemokine C—C motif ligand 2 (CCL2) and decorin (DCN), two H2O2 related factors, exhibited different expression patterns in keratinocytes cultured at 33℃ and 37℃, respectively. This result is consistent with previous research showing that mice stimulated at 37℃ induced more DRG activation than those stimulated at 33℃.Conclusion H2O2 can potentiate the TRPA1-dependent warm response, and H2O2-related factors in keratinocytes can be affected by warm temperature, suggesting the possible role of H2O2 related factors in warm transduction from keratinocytes to downstream TRPA1 in DRG neurons.]]> 2023/12/22 11:27:02 ZHENG Tian-Yuan,WANG Lei,JIANG Yan and LIU Xiao-Ling 142true Objective For people with impaired central vision, novel artificial optical devices can provide access to simplified images by projecting them onto retina regions out of the macula, thus allowing them to utilize peripheral vision to perceive information. We investigate perceptual characteristics of peripheral visual field and provide clues for the design of implantable optical artificial vision devices.Methods We propose an experimental environment for investigating perceptual characteristics of peripheral vision, in which pattern stimuli of symbols, numerals, and Chinese characters are applied to the subjects, with variables including the size, the color combination, the eccentricity, and the motion controlled. The relation between perceptual capability and the variables are analyzed using graphical methods.Results The perceptual capability declines as the eccentricity increases, forming two appreciable trends, and it is significantly influenced by color combination and size.Conclusion Variable combinations with considerable perception rate are provided. The results may promisingly act as a reference for the implantation of artificial optical devices in eyes, the development of color symbol codes for special communication, etc.]]> 2023/12/22 11:27:14 LIN Yan-Ni,LI Yan-Cheng,GE Song,ZHOU Tian-Hang,DONG Wen-Tian,LIU Zi-Yuan,DANG Wei-Min and XU Sheng-Yong 141true Objective Many studies have shown that a high-sugar diet can do a lot of harm to the health of teenagers. As one of the important windows for human to perceive the world, the visual system plays an important role in teenagers’ perception ability. Many studies have confirmed that high-sugar diet can damage visual function, but the current research on the effects of high-sugar diet on visual function mainly focuses on the front of the visual pathway, focusing on the eye system and the optic ganglion, the posterior of the visual pathway, such as the visual central cortex, has not been reported. In order to understand the effect of high sugar diet on visual function, the effect of high sugar diet on central cortex was studied.Methods In this study, we used the behavioral paradigm of GO/NO-GO orientation discrimination task and in vivo multi-channel electrophysiological techniques to explore the effects of high-sugar diet on the primary visual cortex of young mice.Results Behavioral results showed that young mice that had undergone two months of high-sugar diet had a reduced ability to distinguish orientation. The response characteristics of single neurons in the primary visual cortex (V1) were analyzed by electrophysiological techniques. It was found that the ability of orientation tuning of single neurons in the primary visual cortex decreased in mice fed with high-sugar diet, the response variability of neurons increased and the signal-to-noise ratio (SNR) decreased significantly. We further analyzed the changes of SNR and response variability at population level. The results showed that SNR decreased significantly and response variability increased significantly at population level. The results of neuron correlation analysis showed that the noise correlation of primary visual cortex in the high-sugar diet group was significantly up-regulated.Conclusion These results suggest that a high-sugar diet impairs the ability of young mice to discriminate orientation by affecting the receptive field characteristics of individual neurons in the primary visual cortex, as well as the information processing capacity of neuronal populations.]]> 2023/12/22 11:27:17 XU Long-Ling,ZHOU Yi-Feng and XU Guang-Wei 140true Objective Recently, single-atom nanozyme (SAN) has been widely studied due to its high atom utilization and various enzyme-mimicking activities. However, most of the SANs present low metal-atom densities, which restrict their further application and development. The aim of this study is to prepare SAN with high atomic loading and to systematically investigate its enzyme-like activities, in the hope of providing ideas for the preparation of highly loaded SANs and theoretical support for the application of SANs in a wider range of fields.Methods In this work, high-loading Cu SAN with metal contents up to 7.66% (w/w) was prepared using in-situ anchoring strategy. The metal salt precursors were confined into the carbon carriers consisting of aminated graphene quantum dots, and then urea was introduced to provide a rich N-coordination environment for Cu atoms. The chemical bond between Cu atoms and the carrier was further stabilized by pyrolysis under the protection of Ar gas. The low-loading Cu SAN used for comparison was prepared according to traditional metal organic framework (MOFs) anchoring method. Then, the peroxidase (POD)-like, oxidase (OXD)-like and superoxide dismutase (SOD)-like catalytic activities of high/low-loading Cu SAN were tested and compared with 3,3",5,5"- tetramethylbenzidine (TMB) and nitrotetrazolium blue chloride (NBT) as chromogenic substrates. The catalase (CAT)-like catalytic activities of high/low-loading Cu SAN were assessed by monitoring the decomposition rate of hydrogen peroxide (H2O2).Results The results suggested that the high-loading Cu SAN possesses the improved catalytic activity and selectivity towards POD-like and SOD-like activities, showing 3.4-fold and 8.88-fold higher than low-loading Cu SAN, respectively.Conclusion This work provides a strategy to synthesize SAN with high metal-atom density. And the results show that atomic regulation can be achieved by adjusting preparation methods which assist to obtain SAN with different catalytic selectivity, and lay foundation for the development and application of SAN in detection and sensing, diseases treatment and environmental protection.]]> 2023/11/22 0:00:00 CHEN Xin-Zhu,LIU Xiao-Yu,ZHANG Shao-Fang,MU Xiao-Yu and ZHANG Xiao-Dong 139true Objective OxLDL can increase Plin2 expression, then promote the formation of foam cells. LOX1 was a scavenger receptor for oxLDL. Here, we investigate the relationship between Plin2 and LOX1 in the progress of atherosclerosis.Methods The data GSE43292 from GEO database were analyzed for Plin2 and LOX1 expressions and the correlation between Plin2, LOX1 and NF-κB pathway. RAW264.7 cells stimulated by oxLDL served as a cellular model of atherosclerosis. The Plin2, LOX1 and p-p65 expressions were analyzed by immunoblotting, the intracellular lipids were detected by BODIPY 493/503 staining.Results The Plin2 and LOX1 expressions in atheroma plaques were significantly higher than that in adjacent carotid tissues by analyzing GSE43292. The expressions of Plin2 and LOX1, the lipid droplets were increased obviously in RAW264.7 cells after treated with oxLDL for 24 h. And Plin2 overexpression significantly increased the expression of LOX1. This change was more obvious after oxLDL incubation. But knockdown Plin2 maked no difference on LOX1 when without oxLDL treatment. Furthermore, the GSEA plot showed that the expressions of Plin2 and LOX1 were positively related with NF-κB activation in atherosclerosis. Meanwhile, although oxLDL incubation, NF-κB inhibitor JSH-23 pretreatment significantly reduced Plin2 and LOX1 expressions and the amounts of intracellular lipids. In addition, the expressions of Plin2 and LOX1 were significantly inhibited by JSH-23 in spite of Plin2 overexpression plus oxLDL incubation.Conclusion Altogether, Plin2 can promote the intracellular lipids accumulation and may participate in pathophysiological process of atherosclerosis by increasing the expression of LOX1, which at least partly through the activation of NF-κB pathway.]]> 2023/11/22 14:29:45 LIU Qing-Nan,ZHAO Xiao-Hui,LI Shuai,GUO Dong-Ming,YUAN Zhong-Hua,LI Jing,DONG Shan-Shan,WANG Yi-Qian and DAI Zhi-Bing 138true Objective To illustrate the interaction process and mechanism of chemokine CCL11 with receptor CCR3 and glycosaminoglycans (GAGs) and shade light on the regulation mechanism of CCL11-GAGs-CCR3.Methods A stably transfected CCR3-EGFP CHO cell line with single-molecule expression level was constructed using genetic engineering. The interaction between GAGs and CCL11 under different in vitro solution was studied by total internal reflection fluorescence (TIRF) and isothermal titration calorimetry (ITC) technique. The regulation of GAGs-CCL11 on chemotactic behavior of CCR3-EGFP stable cells and the effect of CCR3-EGFP aggregation on cell membrane were investigated by chemotactic assay and living cell monomolecular imaging assay.Results With the increase of chondroitin sulfate chain length, the binding heat release of chondroitin and CCL11 increased, indicating that the interaction force was enhanced and the promotion effect of CCL11 aggregation was enhanced. Single molecule fluorescence imaging technology combined with chemotactic test showed that different types and proportions of GAGs could affect the interaction between CCL11 and CCR3. The addition of GAGs inhibited the chemotactic effect of CCL11 on CCR3-EGFP stable cells and promoted the aggregation ability of CCR3-EGFP. With the increase of molecular mass of chondroitin sulfate, the inhibitory effect was significantly enhanced.Conclusion GAGs can affect the aggregation state of CCL11 and its interaction with receptor CCR3. Our study provides a theoretical basis for further elucidating the interaction between CCL11-GAGs-CCR3.]]> 2023/11/22 0:00:00 LI Guan-Lin,LIU Heng-Heng,DING Yan-Zhi,LI Ji-Qiang and GE Bao-Sheng 137true Objective Aging is an increasingly serious social problem. The cognitive function of the elderly, such as attention, shows a significant decline. Investigating the deficits of visual attention regulatory network caused by aging will further our understanding of the neural mechanism of cognition declines of the elderly, and providing us the theoretical basis to find potential intervention methods.Methods In the study, we used a classical two-target attention task. Subjects were asked to keep their gaze on the central fixation cross, a pair of spots appeared separately located on left and right 13.5° from the cross. After 800-1 200 ms fixation, one of the two spots randomly changed its form or both remained unchanged. We recorded neural activity of young adults and old adults during the attention task using electroencephalogram (EEG).Results Through comparing the differences of EEG activity under visual stimuli changed and unchanged conditions, we found that neural activity of channels in frontal, parietal and temporal areas of young adults significantly changed with the change of stimuli, while neural activity of old adults didn’t follow the change of stimuli.Conclusion These results demonstrated that brain network of the elderly could not effectively react to the change of external visual information load in the attention task. Aging is accompanied by the functional decline of the visual attention regulatory network. Besides, decline of this brain network also showed sex difference. Our research provided new evidence for the decline of visual attention regulatory network of the elderly.]]> 2023/11/22 14:31:03 CAO Dan-Dan,LIU Zu-Xiang,LAI Zhao-Yu,Lü Zhao,SHU Wei,WANG Huan and YANG Yan 136true Objective The cochlear microphonic (CM) signal produced by the bat cochlear outer auditory hair cells contains information related to the movement of the cochlear basilar membrane. The aim of this study was to investigate the mechanism of bat cochlear nerve influence on echolocation by analyzing the relationship between frequency modulation (FM) bat CM signals and their received acoustic frequencies.Methods The CM signals of FM bats (Eptesicus and Pipistrellus) were recorded and analyzed in response to pure acoustic (tone burst) stimulation by placing a set of metal electrodes in the ventral cochlear nucleus.Results Two different species of FM bats (Eptesicus and Pipistrellus) exhibited CM mean frequency response curves during high sound pressure stimulation with a distinct concavity in the narrower specific frequency band vocalization. For Eptesicus, the CM RMS frequency response curves showed significant troughs on both sides of the terminal frequency (TF) of the first and second harmonics; moreover, these troughs were repeated at 15 kHz intervals from 15 kHz onwards; for Pipistrellus, the CM RMS frequency response curves showed significant troughs only on both sides of the first harmonic. The valleys are evident on both sides.Conclusion The relationship between the inhibition exhibited by the CM signal and the FM bat’s first and second harmonic TF reflects the fact that the cochlear nerve produces inhibition for specific frequencies, and this inhibition helps the bat to extract target localization information modulated to the vicinity of the TF.]]> 2023/11/22 14:31:12 MA Zhi-Qiang,KONG You-Hua,GAO Chao,REN Guang-Fu and MA Xin 135true Objective East Asia harbors a vast territory, with many populations and a diversity of languages. China has 34 provincial-level administrative districts which distributed in seven geographical divisions, mainly populated by seven linguistic family-speaking populations. Previous studies have focused on the genetic history of origin, migration and fusion of East Asia populations. We collected and analyzed high-density SNP data of 5 147 individuals in the world, studied the genetic relationship and structure between East Asia populations, especially Chinese populations and other world populations from the perspective of geography and language.Methods We collected and carried out quality control of high-density SNP data of 5 147 individuals in the world. We studied the genetic structure of Chinese population. The final obtained 32 789 SNPs were statistically tested by allele frequency difference analysis. Meanwhile, we employed principal component analysis, phylogenetic tree, ancestry component analysis and D-test statistics to explore the genetic relationship between East Asia populations and other populations in the world, as well as the genetic relationship and structure of Chinese populations.Results We found that there were significant differences among East Asian, African, American and European. Chinese population can be divided into seven subgroups. The genetic clustering in different populations has a strong correlation with their geographical distribution, linguistic families and ethnic origin history.Conclusion We studied the genetic relationship and differences between Chinese population and world population, and systematically studied the genetic substructure of Chinese population. This will enrich the research foundation of population genetics and forensic genetics of East Asia population and provide data support for individualized medical work.]]> 2023/11/22 0:00:00 GU Jia-Qi,JIANG Li,XU Jing-Yi,WANG Han,WEI Yi-Liang and LI Cai-Xia 134true Objective The high density SNP genotype data were obtained by whole genome sequencing (WGS), the accuracy of genotype was evaluated, and this study is intended to establish the method of using whole genome sequencing data for SNP kinship relationship prediction.Methods The samples were sequenced at a depth of 30× through the MGISEQ-200RS sequencing platform, and 645 199 autosomal SNP in the Wegene GSA DNA microarray were extracted from the sequencing data. After quality control and filtering, the prediction relationship was calculated and predicted by IBS/IBD algorithm, and the ancestry of the samples was analyzed.Results The coincidence rate between SNP genotype extracted from sequencing data and Wegene GSA genotype is more than 99.62%.The SNP genotype obtained by sequencing can predict kinship from level 1 to level 4 by using IBS algorithm, and confidence interval accuracy of level 4 kinship prediction is 100%. Using IBD algorithm, the confidence interval accuracy of level 1 to 7 kinship prediction is 100%. The pedigree inference ability of SNP obtained from high-depth whole genome sequence data is not significantly different from that of DNA microarray prediction.At the same time, the use of whole genome sequencing data for ancestry inference is consistent with the survey results.Conclusion The whole genome resequencing technique can be applied to SNP genealogy inference to provide clues for case detection.]]> 2023/11/22 14:32:10 XIE Quan,GUI Xu,ZHAO Wen-Ting,FANG Zhi-Xiao,XU Jing-Yi,LIU Jing and LI Cai-Xia 133true Objective Based on bioelectrical impedance spectroscopy (BIS), we propose a label-free method for real-time detection of biological cell activity. The method determines whether the cells are active or not based on the differences in electrical properties of the cells at different concentrations, physiological and pathological states. It is used to assist physicians to quickly and precisely locate the scalded tissues in patients and achieve effective excision during clinical surgery.Methods Active zebrafish embryonic stem cells were used to simulate human scalded tissues, and bioimpedance spectroscopy was used to identify the physiological state of the cellular tissues.Results There were significant differences in the impedance amplitude changes of the cells in different states, from which it could be found that the impedance amplitude of the active cells at the same concentration was on average 17.25% higher than that of the dead cells, and the relaxation frequency of the active cells occurred 25% earlier than that of the dead cells.Conclusion The experimental data showed that the bioimpedance spectroscopy method can effectively distinguish two types of physiological states of embryonic cells; according to the clustering region, it can be seen that the BIS assay has obvious ability to distinguish cell activity and concentration, which can quickly assist physicians to complete the detection of scalded tissues of patients theoretically.]]> 2023/11/22 14:32:29 XU Kai-Jie,YE Xia,DING Li,HU Song-Pei and YAO Jia-Feng 132true Objective To investigate the effect of 8-week aerobic exercise on the improvement of core behaviors of male and female autistic mice induced by valproic acid (VPA).Methods Experimental mice were randomly divided into the control group (CTL), VPA-induced autism group (VPA) and VPA+aerobic exercise group (VEX), with 10 mice in each group. The pregnant mice were injected with VPA intraperitoneally at E12.5, and their offspring were used as autistic mice. Pups were weaned 28 d after birth and began an 8-week aerobic exercise intervention. The day after exercise, mice were tested in behavioral experiments to detect exploratory behavior, social skills, repetitive stereotypic behavior, cognitive ability and mood. The mice were tested for social skills, repetitive stereotyped behaviors, cognitive and learning memory abilities, exploratory behaviors, and emotions by behavioral assays on the following day after the exercise.Results Both male and female mice in the CTL group showed a significant decrease in the total distance and percentage of time spent in the interaction zone in the 4th socialization compared to the 1st socialization (P<0.01); the total distance and percentage of time spent in the interaction zone in the 5th socialization was significantly increased compared to the 4th socialization (P<0.01); in VPA group, both male and female mice showed no significant change in the total distance and percentage of time spent in the interaction zone in the 4th and 5th socialization; in the VEX group, the total distance and percentage of time spent in the interaction zone by male mice in the 4th socialization was significantly decreased compared to the 1st socialization (P<0.01, P<0.05); and in the VEX group the total distance and percentage of time spent in the social interaction zone by both male and female mice in the 5th socialization was significantly increased compared to the 4th socialization (P<0.01, P<0.05). The results of the first phase of three-box socialization experiment showed that male and female mice in the CTL group spent more time socializing with their social partners than in contact with the empty cages (P<0.01); there was no difference in the time spent by male and female mice in the VPA group in socializing with their social partners and the empty cages; and male and female mice in the VEX group spent a longer time socializing with their social partners (P<0.01). The results of the second phase of three-box test showed that male and female mice in the CTL group showed a significant tendency to socialize with new social partners (P<0.01), whereas no significant changes were observed in the mice of VPA group; aerobic exercise significantly ameliorated this deficit in male and female mice with autism. Compared with the CTL group, VPA-induced significant decreases were observed in the total distance freely moved in the central area of the open field, the time and percentage of time spent in the open arm of cross maze, and the distance and time spent in the white box in both male and female autistic mice (P<0.01); a significant increase in the number of bead burials and time spent in self-grooming (P<0.01); a significant decrease in the cognitive index (P<0.01); a significantly longer latency to find the platform, and significantly decreased the percentage of time spent in the target quadrant and the number of times they traversed the platform (P<0.01). Compared with the VPA group, after 8 weeks of aerobic intervention, male and female mice in the VEX group showed a significant increase in total distance, open-arm dwell time, and percentage of free movement in the central area of the empty field (P<0. 05), and a trend toward a decrease in the dwell time of females in the white box was not significant, the number of beads burying and the time of self-combing were significantly lower (P<0.01, P<0.05); and a significant increase in cognitive index (P<0.05), a significantly shorter time to find the platform, and significantly increased percentage of time spent in the target quadrant and the number of times they traversed the platform (P<0.01), showing excellent learning memory.Conclusion Autistic mice severely suffer from social and cognitive impairments, repetitive stereotyped behaviors, decreased activity level, and the exhibition of anxiety. 8 weeks of aerobic exercise can improve the social and cognitive abilities, alleviate the stereotyped repetitive behaviors, increase the activity level, and positively regulate the anxiety in autistic mice. It is hypothesized that aerobic exercise has an important role in motor rehabilitation of autism, in order to provide a theoretical basis for clinical research.]]> 2024/6/27 18:45:12 LIU Niu,XUE Ya-Qi,WANG Shi-Jiao and ZHEN Zhi-Ping 131true Objective To investigate the effect of mucin 1 (MUC1) on the proliferation and apoptosis of nasopharyngeal carcinoma (NPC) and its regulatory mechanism.Methods The 60 NPC and paired para-cancer normal tissues were collected from October 2020 to July 2021 in Quanzhou First Hospital. The expression of MUC1 was measured by real-time quantitative PCR (qPCR) in the patients with PNC. The 5-8F and HNE1 cells were transfected with siRNA control (si-control) or siRNA targeting MUC1 (si-MUC1). Cell proliferation was analyzed by cell counting kit-8 and colony formation assay, and apoptosis was analyzed by flow cytometry analysis in the 5-8F and HNE1 cells. The qPCR and ELISA were executed to analyze the levels of TNF-α and IL-6. Western blot was performed to measure the expression of MUC1, NF-кB and apoptosis-related proteins (Bax and Bcl-2).Results The expression of MUC1 was up-regulated in the NPC tissues, and NPC patients with the high MUC1 expression were inclined to EBV infection, growth and metastasis of NPC. Loss of MUC1 restrained malignant features, including the proliferation and apoptosis, downregulated the expression of p-IкB、p-P65 and Bcl-2 and upregulated the expression of Bax in the NPC cells.Conclusion Downregulation of MUC1 restrained biological characteristics of malignancy, including cell proliferation and apoptosis, by inactivating NF-κB signaling pathway in NPC.]]> 2024/9/19 20:43:11 WU Shou-Wu,LIN Shao-Kun,NIAN Zhong-Zhu,WANG Xin-Wen,LIN Wei-Nian,ZHUANG Li-Ming,WU Zhi-Sheng,HUANG Zhi-Wei,WANG A-Min,GAO Ni-Li,CHEN Jia-Wen,YUAN Wen-Ting,LU Kai-Xian and LIAO Jun 130true Objective The absorption of substances into blood is mainly dependent on the mesenteric lymphatic pathway and the portal venous pathway. The substances transported via the portal venous pathway can be metabolized by the biotransformation in the liver. On the contrary, the substances in the mesenteric lymph fluid enter the blood circulation without biotransformation and can affect the body directly. Alcohol consumption is strongly linked to global health risk. Previous reports have analyzed the changes of metabolites in plasma, serum, urine, liver and feces after alcohol consumption. Whether alcohol consumption affects the metabolites in lymph fluid is still unknown. Therefore, it is particularly important to explore the changes of substances transported via the mesenteric lymphatic pathway and analyze their harmfulness after alcohol drinking.Methods In this study, male Wistar rats were divided into high, medium, and low-dosage alcohol groups (receiving Chinese Baijiu at 56%, 28% and 5.6% ABV, respectively) and water groups. The experiment was conducted by alcohol gavage lasting 10 d, 10 ml·kg-1·d-1. Then mesenteric lymph fluid was collected for non-targeted metabolomic analysis by using liquid chromatography-mass spectrometry (LC-MS) and bioinformatic analysis. Principal component analysis and hierarchical clustering were performed by using Biodeep. Meanwhile, KEGG enrichment analysis of the differential metabolites was also performed by Biodeep. MetaboAnalyst was used to analyze the relationship between the differential metabolites and diseases.Results The metabolites in the mesenteric lymph fluid of the high-dosage alcohol group change the most. Based on the KEGG enrichment analysis, the pathways of differential metabolites between the high-dosage alcohol group and the control group are mainly enriched in the central carbon metabolism in cancer, bile secretion, linoleic acid metabolism, biosynthesis of unsaturated fatty acids, etc. Interestingly, in the biosynthesis of unsaturated fatty acids category, the content of arachidonic acid is increased by 7.25 times, whereas the contents of palmitic acid, oleic acid, stearic acid, arachidic acid and erucic acid all decrease, indicating lipid substances in lymph fluid are absorbed selectively after alcohol intake. It’s worth noting that arachidonic acid is closely related to inflammatory response. Furthermore, the differential metabolites are mainly related with schizophrenia, Alzheimer’s disease and lung cancer. The differential metabolites between the medium-dosage alcohol and the control group were mainly enriched in phenylalanine metabolism, valine, leucine and isoleucine biosynthesis, linoleic acid metabolism and cholesterol metabolism. The differential metabolites are mainly related to schizophrenia, Alzheimer’s disease, lung cancer and Parkinson’s disease. As the dose of alcohol increases, the contents of some metabolites in lymph fluid increase, including cholesterol, L-leucine, fumaric acid and mannitol, and the number of metabolites related to schizophrenia also tends to increase, indicating that some metabolites absorbed by the intestine-lymphatic pathway are dose-dependent on alcohol intake.Conclusion After alcohol intake, the metabolites transported via the intestinal-lymphatic pathway are significantly changed, especially in the high-dosage group. Some metabolites absorbed via the intestinal-lymphatic pathway are dose-dependent on alcohol intake. Most importantly, alcohol intake may cause inflammatory response and the occurrence of neurological diseases, psychiatric diseases and cancer diseases. High-dosage drinking may aggravate or accelerate the occurrence of related diseases. These results provide new insights into the pathogenesis of alcohol-related diseases based on the intestinal-lymphatic pathway.]]> 2024/9/19 20:43:15 ZHANG Yuan,MENG Zi-Ye,LI Wen-Bo,JING Yu-Meng,LIU Gui-Chen,HAO Zi-Yao,XU Xiu and ZHAO Zhen-Ao 129true Objective Hemoglobin is the iron-containing protein in the red blood cells of many animals. The primary function of hemoglobin is to transport oxygen from lung to tissues. It is composed of two identical α-globin subunits and two identical β-globin subunits. Hemoglobin has unique magnetic properties. The paramagnetism of deoxyhemoglobin, and the diamagnetism of oxyhemoglobin and carboxyhaemoglobin have been reported previously. Studies have also shown that external magnetic field affected blood flow rate, but whether magnetic field may affect the oxygenation rate of hemoglobin remains unknown. Here in this study, we are aiming to address this question with recombinant hemoglobin. Human hemoglobin and yak hemoglobin were selected as the research objects, and a recombinant protein expression and purification system was established to explore the magnetic field effects on the oxygenation rate of hemoglobin, as well as the differences in the oxygenation rate between human hemoglobin and yak hemoglobin under external magnetic field.Methods The recombinant expression and purification system of human and yak hemoglobin was established. The recombinant hemoglobin expression was further optimized and appropriated inducing temperature and IPTG concentration were screened. Recombinant human hemoglobin and yak hemoglobin were purified to homogeneity by affinity chromatography and further by size-exclusion chromatography. SDS-PAGE was used to validate the purification, and UV-Vis spectrum and EPR were used to characterize the biochemical properties of recombinant hemoglobin. Deoxyhemoglobin of human and yak were placed under 0.3 T external magnetic field to test the magnetic field effects on oxygenation rate, and geomagnetic field condition was used as a sham control. The UV-Vis spectrum data were measured every 10 min, and the concentration and proportion of oxygenated hemoglobin, deoxyhemoglobin and methemoglobin were calculated to analyze the effects of magnetic field on the oxygenation rate of hemoglobin. The magnetic properties of human oxygenated hemoglobin and human deoxygenated hemoglobin have been measured by SQUID, a superconducting quantum interference magnetic measurement system. Three biological replications were performed for each experiment. The possible mechanism of the effect of magnetic field on the oxygenation rate of hemoglobin has been investigated and discussed.Results Human and yak hemoglobin were successfully expressed and purified by E.coli prokaryotic expression system. The optimal expression temperature was 30℃, and the most suitable IPTG concentration was 1 mmol/L. EPR results suggested that trace amount of methemoglobin existed both in the purified human hemoglobin and yak hemoglobin proteins. The oxygenation rate of yak hemoglobin appeared to be faster than that of human hemoglobin, and the additional magnetic field treatment significantly increased the oxygenation rate of both human and yak hemoglobin, and yak hemoglobin was more sensitive to magnetic field than human hemoglobin. The paramagnetism of deoxyhemoglobin was verified by SQUID measurement. However, the diamagnetism of oxygenated hemoglobin remains uncertain, probably due to the presence of trace amount of methemoglobin in the sample of oxygenated hemoglobin, which was consistent with EPR results.Conclusion In this study, human and yak hemoglobin were successfully expressed and purified. The purified hemoglobin proteins have similar function and conformational states as native protein. External static magnetic field facilitates hemoglobin oxygenation, and yak hemoglobin seems more sensitive to magnetic field compared with human hemoglobin. These findings provide theoretical basis for the potential applications of applying magnetic field to improve hypoxia symptoms in clinical practice in the future.]]> 2024/9/19 20:43:20 LIU Yan,FANG Yan-Wen,ZHANG Xin and XIE Can 128true Objective Spatial working memory (SWM) is an important function in cognitive behavior, and working memory impairment can seriously affect the patient’s life and cause great stress to the patient. Intermittent theta burst stimulation (iTBS) has been shown to regulate working memory function by entrainment of neural oscillations in different frequencies of the brain, but its regulation of working memory-related neural oscillations and their synchronization is not clear. The purpose of this study was to study the effect of iTBS on neural oscillation and synchronization in local and transbrain regions of rats, and to explore the mechanism of iTBS in regulating working memory.Methods Twenty-four rats were randomly divided into four groups according to their age and whether they received iTBS stimulation (AS: adult stimulation group, AC: adult control group, ES: elderly stimulation group, EC: elderly control group). Using the methods of time-frequency distribution, phase synchronization and phase-amplitude coupling analysis, the changes of local field potential signal neural oscillations in the prefrontal and hippocampal brain regions of theta and gamma bands in the process of spatial working memory behavioral tasks in each group of rats were compared and analyzed, and the relationship between the changes of neural oscillations in the two brain regions and the changes in spatial working memory ability of rats was judged based on the Pearson correlation coefficient.Results With the increase of age, the time taken by the elderly rats to learn the spatial working memory task rules increased significantly (P=0.005 6), and the time taken by iTBS stimulation to learn the SWM task rules in adult rats (P=0.001 1) and elderly rats (P=0.009 0) was shortened. At the same time, compared with adult rats, the time-frequency energy of theta and gamma band neural oscillations in the prefrontal and hippocampal brain regions of elderly rats (theta: P<0.000 1; gamma: P<0.000 1) and phase-amplitude coupling across brain regions (PFC-HPC: P=0.000 2; HPC-PFC: P=0.027 7) decreased to a certain extent, and iTBS stimulation could increase the time-frequency energy of neural oscillations of adult rats (theta: P<0.000 1; gamma: P<0.000 1) and elderly rats (theta: P=0.014 4; gamma: P=0.000 6) and the phase-amplitude coupling effect across brain regions in elderly rats (PFC-HPC: P=0.018 0; HPC-PFC: P=0.022 1). In addition, the time-frequency energy and phase-amplitude coupling of signals in each frequency band of the two brain regions were positively correlated with the behavioral accuracy of rats, while the phase synchronization of theta band and gamma band neural oscillations in the two brain regions during working memory was not correlated with the behavioral accuracy.Conclusion iTBS can enhance SWM ability and cognitive function in elderly rats, and this improvement is associated with increased coupling of time-frequency energy and cross-brain phase amplitude of neural oscillations across theta and gamma bands during SWM tasks. Similarly, in adult rats, iTBS enhances SWM ability and cognitive function by increasing the time-frequency energy of theta and gamma band neural oscillations in both brain regions during SWM tasks. Furthermore, in addition to the main findings, this study provides evidence supporting the state-dependent effects of iTBS stimulation to some extent.]]> 2024/9/19 20:43:23 GUO Miao-Miao,ZHANG Biao,ZHANG Tian-Heng,WANG Tian,ZHAI Hao-Di and XU Gui-Zhi 127true Objective Angle-closure glaucoma (ACG) is one of the major eye-blinding diseases. To diagnose ACG, it is crucial to examine the anterior chamber angle. Current diagnostic tools include slit lamp gonioscopy, water gonioscopy, ultrasound biomicroscopy (UBM), and anterior segment optical coherence tomography (AS-OCT). Slit lamp and water gonioscopy allow convenient observation of the anterior chamber angle, but pose risks of invasive operation and eye infections. UBM can accurately measure the structure of the anterior chamber angle. However, it is complex to operate and unsuitable for patients, who have undergone trauma or ocular surgery. Although AS-OCT provides detailed images, it is costly. The aim of this study is to explore a non-invasive, non-destructive optical reflection tomography (ORT) technique. This technique can achieve low-cost three-dimensional imaging and full-field anterior chamber angle measurement of the porcine eye.Methods The experiment involved assembling an optical reflection tomography system, which included a complementary metal oxide semiconductor (CMOS) camera, a telecentric system, a stepper motor, and a white light source, achieving a spatial resolution of approximately 8.5 μm. The process required positioning the porcine eye at the center of the field of the imaging system and rotating it around its central axis using a stepper motor. Reflection projection images were captured at each angle with an exposure time of 1.0 ms and an interval of 2°. The collected reflection-projection data were processed using a filtered reflection tomography algorithm, generating a series of two-dimensional slice data. These slices essentially represented cross-sectional views of the three-dimensional structural image, and were reconstructed into a complete three-dimensional structural image. Based on the reconstructed three-dimensional structural image of the porcine eye, the anterior chamber angles at different positions were measured, and a distribution map of these angles was drawn. Simultaneously, the ORT measurements were compared with the standard results obtained from optical coherence tomography (OCT) to assess the accuracy of ORT measurements.Results In this study, we successfully obtained the reflection projection data of a porcine eye using ORT technology, reconstructed its three-dimensional structural image, and measured the anterior chamber angle, generating the corresponding distribution map. To better distinguish the different structural parts of porcine eye, the three-dimensional structural image was marked with blue, green, and yellow dashed lines from the outer to the inner layers. The area between the blue and green dashed lines corresponded to the sclera. The area between the green and yellow dashed lines corresponded to the iris. The area inside the yellow dashed line corresponded to the pupil. The three-dimensional structural image clearly revealed the key anatomical features of the porcine eye. It was able to measure the anterior chamber angle at different positions. Additionally, the anterior chamber angle measurements of the porcine eye using ORT were compared with the measurements obtained using a TEL320C1 type OCT system, showing an average deviation of 0.51° and a mean square error of 0.317.Conclusion ORT is a non-invasive, non-destructive, low-cost, and high-resolution imaging technique capable of achieving three-dimensional structural imaging and full-field anterior chamber angle measurement of a porcine eye. This technology offers a new perspective for the diagnosis of angle-closure glaucoma and is significant for the screening, diagnosis, and monitoring of eye diseases, potentially benefiting clinics and small hospitals in remote areas in the future.]]> 2024/9/19 20:43:28 LIU Bi-Wang,ZHONG Jun-Ping,LIN Hai-Na,ZENG Ya-Guang,YU You-Ping,LI Hong-Yi,HAN Ding-An and CHEN Jin-Ying 126true Objective To simulate the microstructure and mechanical properties of tendon tissue and promote its regeneration and repair, electrospinning technology was used to prepare L-polylactic acid (PLLA) fiber membranes loaded with different nano zinc oxide contents and with oriented structures. Physical and chemical characterization and biological performance evaluation were carried out to explore their effects on tendon cell proliferation and differentiation.Methods Preparation of PLLA fiber scaffolds and PLLA/ZnO fiber scaffolds containing different mass fractions of nano ZnO was performed using electrospinning technology. The physicochemical properties of the scaffold were characterized by scanning electron microscopy, mechanical stretching, and EDS spectroscopy. The scaffold was co-cultured with mouse tendon cells to detect its biocompatibility and regulatory effects on cell differentiation behavior.Results The fiber scaffolds were arranged in an oriented manner, and zinc elements were uniformly distributed in the fibers. The tensile strength and Young’s modulus of PLLA/0.1%ZnO fiber scaffolds were significantly higher than PLLA fiber scaffolds. The number of cells on the surface of PLLA/0.1%ZnO fiber scaffold was significantly higher than that of the PLLA group, and the activity was better; mouse tendon cells exhibit directional adhesion and growth along the fiber arrangement direction.Conclusion The oriented PLLA/0.1%ZnO fiber scaffold had excellent physicochemical properties, which can significantly promote the oriented growth, proliferation differentiation of tendon cells. It is expected to be used for tendon tissue regeneration and repair in the future.]]> 2024/8/15 12:35:05 FAN Jia,GU Peng-Cheng,CHENG Xi-Ting,JIANG Qiang,ZHAO Ya,PAN Xiao-Fang and BAI Yan 125true Objective β-Site APP cleaving enzyme 1 (BACE1) is a rate-limiting enzyme involved in the formation of amyloid plaques in Alzheimer’s disease (AD), and its expression and activity play a crucial role in the development of AD. The interacting protein of BACE1 can directly or indirectly regulate BACE1 in the transcription, translation, modification, intracellular transport and other links of BACE1 by directly binding, indirectly binding, and participating in various cell signal transduction pathways, so as to participate in the occurrence of AD and the process of disease. This study aimed to screen and validate the interacting proteins of BACE1, providing new insights into the mechanisms of amyloid plaque formation.Methods Co-immunoprecipitation (Co-IP) and mass spectrometry (MS) were used to enrich and identify BACE1 interacting proteins in the hippocampus of wild type (WT) mice and AD model mice. For candidate BACE1 interacting proteins, GO enrichment analysis and KEGG pathway enrichment analysis were used to explore the subcellular localization, molecular function, participating biological processes and participating signaling pathways of BACE1 interacting proteins. The protein-protein interaction (PPI) network of BACE1 was further constructed to explore the potential proteins interacting with BACE1. By searching the mouse genomeinformation (MGI) website and NCBI database, the more reliable proteins among the potential BACE1 interacting proteins were screened. Co-IP assay and immunofluorescence confocal technology were used to preliminarily verify the interaction between the proteins, and the changes in protein expression levels of the interacting proteins in AD cell models were explored.Results A total of 614 differentially expressed proteins interacting with BACE1 were identified in AD group. GO enrichment analysis showed that the BACE1 interacting proteins in the AD group were mainly located in membrane organelles such as Golgi apparatus, endoplasmic reticulum, endosome, lysosome and vesicles, which had molecular functions such as ion channel regulation, protein kinase activity, transcription factor binding and passive transmembrane transporter activity. It is mainly involved in the biological processes of immune response regulation cell surface receptor signaling pathway, targeting Golgi vesicles transport, circadian rhythm regulation, Purkinje cell layer development, etc. KEGG analysis showed that BACE1 interacting proteins in AD were mainly involved in the PI3K-Akt signaling pathway, mTOR signaling pathway and other neurodegenerative disease-related pathways. The PPI network of BACE1 showed that a total of 12 proteins were identified as high confidence binding proteins, including PRNP, APOE, SYP, NSF, NUMB, SNAP91, HSP90aa1, UCHL1, BIN1, SNX27, Rheb, Ap2m1, of which, NSF, NUMB, SNAP91, HSP90aa1 were newly identified candidate proteins. After further verification, we found that NSF not only interacts with BACE1, but also interacts with amyloid precursor protein (APP), the substrate of BACE1, and the expression level of NSF is up-regulated in the AD cell model constructed by Aβ42 induction.Conclusion BACE1 binding proteins participate in multiple AD-associated biological processes and signal pathways. NSF is a newly identified BACE1 binding protein that interacts with BACE1, and the protein expression level of NSF is up-regulated in the AD cell model. It is predicted that the interaction between NSF and BACE1 is involved in regulating the course of AD, providing a new target and direction for the study of the mechanism of AD.]]> 2024/8/15 0:00:00 LIU Cong-Cong,WANG Ya-Qi and WANG Pei-Chang 124true Objective DnaG primase in Mycobacterium tuberculosis (MtuDnaG) plays a vital role in DNA replication, making it a target for novel antituberculosis drug discovery. However, the mechanism of MtuDnaG priming is not fully understood, which hinders the screening of MtuDnaG inhibitors. In this work, the specific recognition sites (SRS) in ssDNA for MtuDnaG binding was investigated and the interactions between MtuDnaG and ssDNA template was discussed.Methods By biochemical and biophysical methods, the binding of the didomain of MtuDnaG (MtuP49, containing the zinc-binding domain and RNA polymerase domain) to ssDNA template with various trinucleotide sites was evaluated, the affinity of MtuP49 to ssDNA template was measured.Results The present study suggested the 5"-GCG/C-3" as the potential SRS in ssDNA for specific binding to MtuDnaG. Besides, 5"-GCG/C-3" sites were further identified within the oriC region of M. tuberculosis genome. Importantly, the 3" sequence flanking the 5"-GCG/C-3" site markedly affected the binding affinity of ssDNA to MtuP49. Mutagenesis studies showed that substitution of residue Arg31 in the zinc-binding domain affected the binding activity of MtuP49 to template ssDNA. Combined with the predicted structure of MtuP49, an intramolecular rearrangement of zinc-binding domain relative to the RNA polymerase domain was implied to be essential in the binding of MtuP49 to template ssDNA.Conclusion This study firstly identified the SRS in ssDNA for MtuDnaG binding, the key factors affecting MtuDnaG binding to ssDNA was revealed. The above results provide evidence to shed light on the mechanism of MtuDnaG priming, and pave the way for development of novel DnaG-targeted antituberculosis drugs.]]> 2024/8/15 12:35:17 CHEN Jiang,LUO Hao,ZHANG Zhi-Ming,SONG Xu and WANG Gang-Gang 123true Objective Karoshi, death from overwork, is a serious problem with unclear identification standards and mechanisms. This study aims to establish a karoshi rat model by integrating weight-bearing swimming and sleep deprivation. This model will enable us to investigate the adverse effects of acute physical and mental fatigue on cardiac functions and explore the response mechanisms to overwork using integrated omics approaches, specifically metabonomics and proteomics.Methods The experimental design involved healthy male sprague-dawley (SD) rats subjected to weight-bearing swimming and sleep deprivation for 7 d. The rats were monitored for changes in physiological function indexes, including electrocardiogram and respiration. Protein digestion, iTRAQ labeling, and quantitative data analyses were performed to determine differentially expressed proteins (DEPs). Additionally, GC-MS analysis was conducted to identify differential metabolites. The integration analysis of differential metabolites and proteins shared by the fatigue group and the overwork group was performed to construct a relevant metabolic pathway network and integrate the proteomics and metabolomics data. Statistical analysis was carried out using one-way ANOVA and Duncan’s multiple range t-tests.Results The rats subjected to weight-bearing swimming and sleep deprivation showed various physical and behavioral changes associated with fatigue, including hair disorder, decreased muscle tension, reduced food intake, and weight loss. Analysis of cardiac functions revealed cardiac hypertrophy and heart failure in the fatigue and karoshi groups, as evidenced by changes in heart color, myocardial fiber structure, heart rate, respiratory rate, and cardiac ultrasound measurements. Metabolomics analysis using GC-MS identified several differential metabolites in response to overwork, including amino acids involved in various metabolic pathways. Proteomic analysis using iTRAQ technology identified DEPs in the fatigue and karoshi groups, with a subset of DEPs shared by both groups. The GO analysis revealed that the up-regulated DEPs were primarily associated with mitochondria and peroxisomes in the cellular component category. The Reactome analysis further highlighted the enrichment of DEPs in the transfer of ferriheme from methemoglobin to hemopexin pathway. Integration analysis of the DEPs and differential metabolites revealed the activation of autophagy, increased mitochondrial oxidative phosphorylation, enhanced branched-chain amino acid degradation, and altered peroxisomal β-oxidation. These findings suggested complex metabolic adaptations to meet the increased energy demands during overwork while also dealing with oxidative stress. Furthermore, the reprogramming of energy metabolism was observed, with upregulation of fatty acid β-oxidation enzymes and glycolysis-related enzymes in the fatigue group, indicating a shift towards glucose metabolism. In contrast, the karoshi group showed a decreased dependence on fatty acids as an energy source and increased utilization of glucose. The model proposed in this study highlights the interconnected metabolic changes involving mitochondria, peroxisomes, and lysosomes in response to overwork. The findings contribute to our understanding of the mechanisms involved in overwork-related pathologies and provide a basis for further research in the field of karoshi.Conclusion Overall, metabolic reprogramming might provide sufficient energy to the heart, alleviate oxidative stress and damage to cardiac cells in response to excessive exertion and fatigue. Our findings provide an insight into response mechanism to overwork death and lay a foundation for further research on overwork death.]]> 2024/8/15 12:35:43 LI Jia-Min,SU Rui-Bing,YU Xiao-Jun and ZHENG Yong-Xia 122true Objective Sodium hyaluronate (HA) was used as the research object to modify it with phenolic acid in order to obtain the molecular structure with better antioxidant activity or even new activity.Methods In this study, 5 kinds of phenolic acid-sodium hyaluronate was prepared by free radical-mediated grafting method, and the grafts with the highest grafting degree were selected to optimize the synthesis conditions. Then, grafts structure and physicochemical properties were analyzed. The grafts were characterized by IR, UV, 1H NMR, FESEM and TGA spectra. The in vitro antioxidant capacity of grafts was determined by the scavenging ability of DPPH·, ABTS+· and O2-·.Results Among 5 kinds of phenolic acid-sodium hyaluronate, the grafting rate of ferulic acid-sodium hyaluronate copolymer (FA-HA) was highest , which was chosen as experimental sample in the following tests. Firstly, the reaction conditions were investigated and the highest grafting rate was (16.59±0.31) mg/g at the optimal preparation conditions. Then, FA-HA structure and physicochemical properties were analyzed. Data from UV, IR, 1H NMR analyses, TGA showed that FA were successfully grafted to HA. Compared with HA, the results of gel permeation chrematography (GPC) showed that the molecular mass distribution of FA-HA copolymer decreased from 34.4 to 31.5 ku, but the uniformity of molecular distribution was improved. FESEM results showed that the structure of copolymer exhibited a closely connected lamellar structure with a relatively smooth surface. TGA results showed that thermal stability of FA-HA had a little decline. The antioxidant performance in vitro results showed that, during 0.25-10 g/L, FA-HA can eliminate (83.76±4.86)% DPPH·, (76.95±5.06)% ABTS+· and (83.08±2.51)% O2-· respectively at 10 g/L. which were higher than that of native HA and FA.Conclusion FA and HA were successfully grafted together by free radical grafting, and the grafted FA-HA had better antioxidant activity in vitro, which provided a theoretical basis for further research and development of phenolic acid-HA grafts.]]> 2024/8/15 12:35:47 ZHANG Xiao-Yue,WANG Xiao-Na,JIANG Min,HAN Ting-Ting,GONG Jin-Song,LI Qing-Na,YANG Su-Zhen and SHI Jin-Song 121true Objective In recent years, the negative impact of microgravity on astronauts’ nervous systems has received widespread attention. The repetitive transcranial magnetic stimulation (rTMS) technology has shown significant positive effects in the treatment of neurological and psychiatric disorders. The potential benefits of combined frequency stimulation (CFS) which combines different frequency stimulation patterns in ameliorating neurological dysfunctions induced by the microgravity environment, still require in-depth investigation. Exploring the therapeutic effects and electrophysiological mechanisms of CFS in improving various neurological disorders caused by microgravity holds significant importance for neuroscience and the clinical application of magnetic stimulation.Methods This study employed 40 C57BL/6 mice, randomly divided into 5 groups: sham group, hindlimb unloading (HU) group, 10 Hz group, 20 Hz group, and combined frequency stimulation (10 Hz+20 Hz, CFS) group. Mice in all groups except the sham group received 14 d of simulated microgravity conditions along with 14 d of repetitive transcranial magnetic stimulation. The effects of CFS on negative emotions and spatial cognitive abilities were assessed through sucrose preference tests and water maze experiments. Finally, patch-clamp techniques were used to record action potentials, resting membrane potentials, and ion channel dynamics of granule neurons in the hippocampal dentate gyrus (DG) region.Results Compared to the single-frequency stimulation group, behavioral results indicated that the combined frequency stimulation (10 Hz+20 Hz) significantly improved cognitive impairments and negative emotions in simulated microgravity mice. Electrophysiological experiments revealed a decrease in excitability of granule neurons in the hippocampal DG region after HU manipulation, whereas the combined frequency stimulation notably enhanced neuronal excitability and improved the dynamic characteristics of voltage-gated Na+ and K+ channels.Conclusion The repetitive transcranial magnetic stimulation with combined frequencies (10 Hz+20 Hz) effectively ameliorates cognitive impairments and negative emotions in simulated microgravity mice. This improvement is likely attributed to the influence of combined frequency stimulation on neuronal excitability and the dynamic characteristics of Na+ and K+ channels. Consequently, this study holds the promise to provide a theoretical basis for alleviating cognitive and emotional disorders induced by microgravity environments.]]> 2024/7/19 14:48:04 ZHAO Jun-Qiao,ZHU Ming-Qiang,ZHU Hai-Jun,FU Rui,ZHANG Ze,WANG Jia-Le and DING Chong 120true Objective This study aimed to identify a potential miRNA-mRNA axis in neurofibromatosis type 2 (NF2)-negative meningiomas, investigate their target relationships, and determine their biological functions.Methods The GSE17792 dataset, which contains data related to NF2-negative meningiomas, was downloaded from the Gene Expression Omnibus (GEO) database. The limma package of R software was used to determine the differentially expressed miRNAs (DeMiRNAs). The miRWalk 2.0 database was applied to obtain the target genes of DeMiRNAs. The Search Tool for the Retrieval of Interacting Genes (STRING) database was utilized to build protein-protein interaction (PPI) networks, and hub genes were identified via Cytoscape software. The expression and biological roles of the screened miRNAs were further validated.Results Altogether, 86 DeMiRNAs, consisting of 52 upregulated and 34 downregulated miRNAs, were found in NF2-negative meningioma tumor samples compared with arachnoid tissue controls. Fourteen miRNAs associated with 274 target genes were identified among these DeMiRNAs, and miRNA-target gene networks were constructed based on these data. Analysis with cytoHubba showed that two miRNAs (hsa-miR-650 and hsa-miR-623) were among the top 20 key hub genes in the PPI network. Further qRT-PCR experimental verification suggested that the expression of hsa-miR-650 was significantly higher in NF2-negative meningiomas than in normal brain tissues. Downregulation of hsa-miR-650 inhibited the proliferation and induced the apoptosis of NF2-negative meningioma cells. Finally, RAC1 was identified as a target of hsa-miR-650.Conclusion Hsa-miR-650 acts as a tumor promoter and might function as a therapeutic target for patients with NF2-negative meningiomas.]]> 2024/7/19 14:48:06 ZHANG Chao,LI Peng,WANG Bo,WANG Ying and LIU Pi-Nan 119true Objective Exosomes are microvesicles which could be secreted by all cell types with diameters between 30 and 150 nm. It was widely distributed in body fluids including blood, urine, and breast milk. Exosomes are considered as potential biomarkers and drug carriers by reason of containing nucleic acids, lipids, proteins and other bioactive molecules. Milk-derived exosomes have been widely used as drug delivery carriers to treat targeted diseases with a lower cost, higher biocompatibility and lower immunogenicity. Until now, there is no research about the milk-derived exosomes phosphorylation to reveal the difference of protein phosphorylation in different species of milk. To investigate the pathways and proteins with specific functions, phosphorylated proteomic analysis of milk-derived exosomes from different species is performed, and provide new ideas for exploring diversified treatments of disease.Methods Whey and exosomes derived from bovine, porcine and caprine milk were performed for proteomics and phosphoproteomics analysis. The relationship between milk exosome proteins from different species and signaling pathways were analyzed using bioinformatics tools.Results A total of 4 191 global proteins, 1 640 phosphoproteins and 4 064 phosphosites were identified from 3 species of milk-derived exosomes, and the exosome proteins and phosphoproteins from different species were significantly higher than those of whey. Meanwhile, some special pathways were enriched like Fcγ-mediated phagocytosis from bovine exosomes, pathways related with neural and immune system from caprine exosomes, positive and negative regulation of multiple activities from porcine exosomes.Conclusion In this study, the proteomic and phosphoproteomic analyses of exosomes and whey from bovine, porcine and caprine milk were carried out to reveal the difference of composition and related signaling pathways of milk exosome from different species. These results provided powerful support for the application of exosomes from different milk sources in the field of disease treatment.]]> 2024/7/19 14:48:16 LIU Chang-Mei,HU Yi-Fan,CHEN Wen-Yan,LIU Dan,SHI Jie and YANG Gang-Long 118true Objective Human Ku70 protein mainly involves the non-homologous end joining (NHEJ) repair of double-stranded DNA breaks (DSB) through its DNA-binding properties, and it is recently reported having an RNA-binding ability. This paper is to explore whether Ku70 has RNA helicase activity and affects miRNA maturation.Methods RNAs bound to Ku protein were analyzed by RNA immunoprecipitation sequencing (RIP-seq) and bioinfomatic anaylsis. The expression relationship between Ku protein and miRNAs was verified by Western blot (WB) and quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) assays. Binding ability of Ku protein to the RNAs was tested by biolayer interferometry (BLI) assay. RNA helicase activity of Ku protein was identified with EMSA assay. The effect of Ku70 regulated miR-124 on neuronal differentiation was performed by morphology analysis, WB and immunofluorescence assays with or without Zika virus (ZIKV) infection.Results We revealed that the Ku70 protein had RNA helicase activity and affected miRNA maturation. Deficiency of Ku70 led to the up-regulation of a large number of mature miRNAs, especially neuronal specific miRNAs like miR-124. The knockdown of Ku70 promoted neuronal differentiation in human neural progenitor cells (hNPCs) and SH-SY5Y cells by boosting miR-124 maturation. Importantly, ZIKV infection reduced the expression of Ku70 whereas increased expression of miR-124 in hNPCs, and led to morphologically neuronal differentiation.Conclusion Our study revealed a novel function of Ku70 as an RNA helicase and regulating miRNA maturation. The reduced expression of Ku70 with ZIKV infection increased the expression of miR-124 and led to the premature differentiation of embryonic neural progenitor cells, which might be one of the causes of microcephaly.]]> 2024/7/30 17:10:41 HUANG Ai-Xue,LI Rui-Ting,ZHAO Yue-Chao,LI Jie,LI Hui,DING Xue-Feng,WANG Lin,XIAO Can,LIU Xue-Mei,QIN Cheng-Feng and SHAO Ning-Sheng 117true Methods A total of 23 DOC patients were included, including 11 MCS patients and 12 VS patients. Based on the Oddball paradigm, auditory naming stimulation was performed on DOC patients and EEG signals of DOC patients were synchronously collected. The brain functional networks were constructed using multivariate Granger causality method, and the differences in node degree, clustering coefficient, global efficiency, and causal flow of the brain networks between MCS patients and VS patients were calculated. The differences in network characteristics of patients with different levels of consciousness under auditory stimulation were compared from the perspective of cooperation between brain regions.Results The causal connectivity between most brain regions in MCS patients was stronger than that in VS patients, and MCS patients had more brain network connectivity edges than VS patients. The average degree (P<0.05), average clustering coefficient, and global efficiency (P<0.05) of MCS patients under naming stimulation were higher than those of VS patients. The difference in out-degree between each node of VS patients was larger, and the difference in in-degree between each node of MCS patients was smaller. The difference in in-degree of MCS patients was more significant than that of VS patients, and the inflow and outflow of information in the brain functional network of MCS patients were stronger than those of VS patients. MCS and VS patients had differences of causal flow in the frontal and temporal lobes, the direction of information transmission in the parietal lobe and central region was not the same, and MCS patients had more electrodes as causal sources than VS patients.Conclusion The information transmission ability of MCS patients is stronger than that of VS patients under auditory naming stimulation. Compared with VS patients, MCS patients have an increase in the number of electrode channels as the causal source, an increase in information output to other brain regions, and also an increase in the information output within brain regions, which may indicate a better state of consciousness in patients. MCS patients have more electrode channels for information output in the frontal lobe than VS patients, and the number of electrode channels for changing the direction of information transmission in the frontal lobe is the highest. The frontal lobe is closely related to the level of consciousness in patients with consciousness disorders. This study can provide a theoretical basis for the grading evaluation of consciousness levels in DOC patients.]]> 2024/7/30 17:10:42 YIN Ning,YANG Fan,LI Zhong-Zhen,HAN Ya-Mei,LI Ji-Cheng and XU Gui-Zhi 116true Objective Sorafenib is a first-line only drug approved for the treatment of advanced hepatocellular carcinoma (HCC). Resistance to sorafenib means that treatment outcomes are often unsatisfactory. Although the mechanism underlying sorafenib resistance remains unclear, resistance may occur through Akt signaling pathway activation in HCC. Dihydrotanshinone (DHT), a lipophilic component of traditional Chinese medicine Salvia miltiorrhiza Bunge, has multiple anti-tumor activities and inhibits Akt activation. The effect and mechanism of DHT combined with sorafenib on HCC have not been investigated. In this study, we investigate whether DHT potentiates the anti-cancer activities of sorafenib against HCC.Methods In this study, the effects of sorafenib and DHT on the viability, apoptosis and drug sensitivity of Huh7 and HepG2 cells were verified by Cell Counting Kit-8 (CCK-8) and flow cytometry. Akt, P-Akt, Caspase3, GSK-3β, P-GSK-3β, ribosomal protein S6 kinase (S6K), P-S6K, Cyclin D1, Bcl-xl, Bcl-2, and Bax expression levels were analyzed via Western blot. All data were statistically compared using one-way analysis of variance (ANOVA) and Dunnett test. Statistical analysis using SPSS 20.0 statistical software.Results DHT inhibit proliferation and promote apoptosis in HCC cells by reducing Akt activation. DHT inhibits the expression and activation of Akt downstream factors, including GSK-3β and S6K, which regulate the apoptotic response and are activated and upregulated by sorafenib treatment. Both sorafenib and DHT downregulate cyclin D1 expression and DHT upregulates Bax expression and downregulates Bcl-2 and Bcl-xl expression. However, sorafenib had little influence on Bcl-2 family protein expression.Conclusion DHT may enhance the proliferation inhibition and apoptosis induction of sorafenib in HCC cells by inhibiting the activation of Akt signaling pathway, thus enhancing the anticancer effect of sorafenib.]]> 2024/7/30 17:10:43 WANG Wei,JIA Sheng-Nan,FAN Sun-Fu and XU Li-Shan 115true Objective Temporal heterogeneity in lung cancer presents as fluctuations in the biological characteristics, genomic mutations, proliferation rates, and chemotherapeutic responses of tumor cells over time, posing a significant barrier to effective treatment. The complexity of this temporal variance, coupled with the spatial diversity of lung cancer, presents formidable challenges for research. This article will pave the way for new avenues in lung cancer research, aiding in a deeper understanding of the temporal heterogeneity of lung cancer, thereby enhancing the cure rate for lung cancer.Methods Raman spectroscopy emerges as a powerful tool for real-time surveillance of biomolecular composition changes in lung cancer at the cellular scale, thus shedding light on the disease’s temporal heterogeneity. In our investigation, we harnessed Raman spectroscopic microscopy alongside multivariate statistical analysis to scrutinize the biomolecular alterations in human lung epithelial cells across various timeframes after benzo(a)pyrene exposure.Results Our findings indicated a temporal reduction in nucleic acids, lipids, proteins, and carotenoids, coinciding with a rise in glucose concentration. These patterns suggest that benzo(a)pyrene induces structural damage to the genetic material, accelerates lipid peroxidation, disrupts protein metabolism, curtails carotenoid production, and alters glucose metabolic pathways. Employing Raman spectroscopy enabled us to monitor the biomolecular dynamics within lung cancer cells in a real-time, non-invasive, and non-destructive manner, facilitating the elucidation of pivotal molecular features.Conclusion This research enhances the comprehension of lung cancer progression and supports the development of personalized therapeutic approaches, which may improve the clinical outcomes for patients.]]> 2024/7/30 17:10:44 ZHOU Hai-Tao,YAO Wei,CUI Cao-Zhe,ZHOU Xiao-Tong,LIANG Xi-Long,QIN Cheng-Bing,XIAO Lian-Tuan,WU Zhi-Fang and LI Si-Jin 114true Objective In recent years, with the intensification of environmental issues and the depletion of ozone layer, incidence of skin tumors has also significantly increased, becoming one of the major threats to people’s lives and health. However, due to factors such as high concealment in the early stage of skin tumors, unclear symptoms, and large human skin area, most cases are detected in the middle to late stage. Early detection plays a crucial role in postoperative survival of skin tumors, which can significantly improve the treatment and survival rates of patients. We proposed a rapid non-invasive electrical impedance detection method for early screening of skin tumors based on bioimpedance spectroscopy (BIS) technology.Methods Firstly, we have established a complete skin stratification model, including stratum corneum, epidermis, dermis, and subcutaneous tissue. And the numerical analysis method was used to investigate the effect of dehydrated and dry skin stratum corneum on contact impedance in BIS measurement. Secondly, differentiation effect of different diameter skin tumor tissues was studied using a skin model after removing the stratum corneum. Then, in order to demonstrate that BIS technology can be used for detecting the microinvasion stage of skin tumors, we conducted a simulation study on the differentiation effect of skin tumors under different infiltration depths. Finally, in order to verify that the designed BIS detection system can distinguish between tumor microinvasion periods, we conducted tumor invasion experiments using hydrogel treated pig skin tissue.Results The simulation results show that a dry and high impedance stratum corneum will bring about huge contact impedance, which will lead to larger measurement errors and affect the accuracy of measurement results. We extracted the core evaluation parameter of relaxed imaginary impedance (Zimag-relax) from the simulation results of the skin tumor model. When the tumor radius (Rtumor) and invasion depth (h)>1.5 mm, the designed BIS detection system can distinguish between tumor tissue and normal tissue. At the same time, in order to evaluate the degree of canceration in skin tissue, the degree of tissue lesion (εworse) is defined by the relaxed imaginary impedance (Zimag-relax) of normal and tumor tissue (εworse is the percentage change in virtual impedance of tumor tissue relative to that of normal tissue), and we fitted a Depth-Zimag-relax curve using relaxation imaginary impedance data at different infiltration depths, which can be applied to quickly determine the infiltration depth of skin tumors after being supplemented with a large amount of clinical data in the future. The experimental results proved that when εworse=0.492 0, BIS could identify microinvasive tumor tissue, and the fitting curve correction coefficient of determination was 0.946 8, with good fitting effect. The simulation using pig skin tissue correlated the results of real human skin simulation with the experimental results of pig skin tissue, proving the reliability of this study, and laying the foundation for further clinical research in the future.Conclusion Our proposed BIS method has the advantages of fast, real-time, and non-invasive detection, as well as high sensitivity to skin tumors, which can be identified during the stage of tumor microinvasion.]]> 2024/5/21 20:01:10 PENG Jun-Wen,HU Song-Pei,HONG Zhi-Yang,WANG Li-Li,LIU Kai and YAO Jia-Feng 113true Objective This study aims to explore and elucidate the possible mechanism of action of Shakuyakukanzoto (SKT) in improving ulcerative colitis (UC) in mice through regulating energy metabolism and polarization of macrophages.Methods The mouse UC model was constructed by administering 3% dextran sulfate sodium salt (DSS), and the mice were treated with SKT intragastrically. In addition, single-cell sequencing and enrichment of metabolic pathways against two datasets, GSE21157 and GSE210415, were conducted first. Second, the extraction and metabolomics of peritoneal macrophages from UC mice were verified. Then, the pathway of differentially abundant metabolite enrichment and the correlation of UC risk were analyzed depending on univariate Mendelian randomization of two samples weighted by standard inverse variance. Finally, the results were verified by qRT-PCR, Western blot, and flow cytometry.Results According to the HE staining results, SKT can significantly alleviate colon damage caused by DSS. Macrophages, NK cells, T cells, and more than 10 different types of cells, based on single-cell sequencing analysis, are detected in the intestinal wall. In the disease group, we can conclude that the activity of 49 macrophage metabolic pathways, mainly involved in energy metabolism, is significantly upregulated through a comparison of the two datasets. In energy metabolomics, 10 and 18 types of metabolites accompanied by significantly upregulated and downregulated differential expression were identified in the treatment group and the model group, as well as the model group and the blank group, respectively. Meanwhile, these differentially expressed metabolites present an obvious correlation with glycolysis and oxidative phosphorylation. Moreover, it can be inferred that glycolysis and the oxidative phosphorylation-related gene NDUFS1 (OR: 0.56, 95% CI: 0.48-0.98, P=0.000 068) are associated with a reduced risk of UC based on the univariate Mendelian randomization of two samples weighted based on standard inverse variance. By analyzing the difference in transcription levels between the two datasets, the transcription level of NDUFS1 in UC was decreased compared with that in the normal group. The results of qRT-PCR, Western blot, and flow cytometry indicate that SKT can promote the expression of the oxidative phosphorylation protein NDUFS1 in macrophages and inhibit the M1-type polarization of macrophages. Furthermore, knockdown/overexpression of NDUFS1 can affect the effect of SKT on M1-type polarization of macrophages.Conclusion Based on the results of this study, SKT inhibits macrophage polarization toward the M1 phenotype by regulating the level of the oxidatively phosphorylated protein NDUFS1 in macrophages; hence, UC is also relieved in mice. These conclusions not only reveal the therapeutic mechanism of SKT for UC but also provide a new theoretical basis for clinical application.]]> 2024/5/21 20:01:14 XIAHOU Zhi-Kai,XIAO Hong,SONG Ya-Feng and HAN Jun 112true Objective The purpose of this study was to investigate the effects of transcutaneous electrical acupoint stimulation (TEAS) on cognitive function of vascular dementia (VD) rats and its mechanism.Methods VD rat model was established by modified two-vessel occlusion (2-VO). After modeling, TEAS and electroacupuncture (EA) were used to stimulate Baihui and Zusanli points of rats respectively for 14 d. After treatment, novel object recognition test, Morris water maze test, and Y maze test were used to evaluate the spatial memory and learning ability of rats. Hematoxylin and eosin staining was used to observe the morphology of hippocampal neurons. Transmission electron microscopy was used to observe the ultrastructure of hippocampal mitochondria. Enzyme-linked immunosorbent assay kits were used to detected the levels of SOD, CAT, GSH-Px, MDA and ROS in serum of rats. Western blot was used to detect the expression of PGC-1α, TFAM, HO-1, NQO1 proteins in the hippocampus, Keap1 protein in the cytoplasm and Nrf2, NRF1 proteins in the nucleus.Results After treatment for 14 d, compared to the model group, the escape latency of VD rats decreased, while the discrimination index, the times of rats crossing the original platform area, the residence time in the original platform quadrant, and the percentage of alternation increased. TEAS can improve the structure of hippocampal neurons and mitochondria of VD rats, showing that neurons were arranged more regularly and distributed more evenly, nuclear membrane and nucleoli were clearer, and mitochondrial swelling were reduced, mitochondrial matrix density were increased, and mitochondrial cristae were more obvious. The levels of SOD, GSH-Px and CAT in serum increased significantly, while the concentration of MDA and ROS decreased. TEAS also up-regulated the expression levels of PGC-1α TFAM, NQO1 and HO-1 proteins in the hippocampus and Nrf2, NRF1 proteins in the nucleus, but down-regulated the Keap1 protein in the cytoplasm.Conclusion TEAS can improve cognition, hippocampal neurons and mitochondrial structure of VD rats, and the effect is better than EA. The mechanism may be the activation of PGC-1α mediated mitochondrial biogenesis and antioxidant stress, which also provides a potential therapeutic technology and experimental basis for the treatment of VD.]]> 2024/5/21 20:01:24 KANG Ji-Liang,HU Ke,LU Jun-Yue,HU Zi-Wei,XU Biao-Ping,LI Xiao-Mao,ZHOU Jun-Jie,JIN Yu,TANG Min,XU Rong and WEN You-Liang 111true Objective To explore whether miR-375 regulates the malignant characteristics of osteosarcoma (OS) by influencing the expression of MMP13.Methods Plasmid DNAs and miRNAs were transfected into OS cells and HEK293 cells using Lipofectamine 3000 reagent. Real-time quantitative polymerase chain reaction was performed to measure the expression of miR-375 and MMP13 in OS patients and OS cells. Western blot was performed to analyze the MMP13 protein in the patients with OS and OS cells. The targeting relationship between miR-375 and MMP13 was analyzed by luciferase assay. Migration and invasion were analysed by heal wound and transwell assays, respectively.Results miR-375 expression in OS tissues was lower than that in normal tissues. The expression of MMP13 was upregulated in OS tissues. MMP13 expression was negatively correlated with miR-375 expression in patients with OS. Migration and invasion were significantly inhibited in OS cells with the miR-375 mimic compared with OS cells with the miRNA control. MMP13 partially reversed the inhibition of migration and invasion induced by miR-375 in the OS cells.Conclusion miR-375 attenuates migration and invasion by downregulating the expression of MMP13 in OS cells.]]> 2024/5/21 20:01:36 LIU Zhong,HE Lei,XIAO Jian,ZHU Qing-Mei,XIAO Jun,YANG Yong-Ming,LUO Yong-Jian,MO Zhong-Cheng,ZHANG Yi-Qun and LI Ming 110true Objective Acute myocardial infarction (AMI) is a highly prevalent and deadly disease globally, with its incidence continuing to rise in recent years. Timely reperfusion therapy is crucial for improving the prognosis of AMI patients. However, myocardial reperfusion can lead to irreversible myocardial ischemia/reperfusion (MI/R) injury, which is associated with adverse cardiovascular outcomes following AMI. Studies have shown that microRNAs (miRNAs) are abnormally expressed during MI/R injury and play an important role in the fate of cardiomyocytes. Effective preventive and therapeutic strategies against MI/R injury remain lacking in clinical practice, necessitating elucidation of the molecular mechanisms underlying MI/R onset and progression. This study investigated the role of microRNA-878 (miR-878) in the regulation of mitochondria-mediated apoptosis in MI/R injury.Methods The H9c2 cells were flushed with a gas mixture containing 1% O2, 5% CO2 and 94% N2 for 3 h. Then the cells were incubated in complete culture medium under 5% CO2 and 95% air for 6 h to mimic in vivo hypoxia/reoxygenation (H/R) injury. Cell viability were detected by CCK-8 assay. The concentrations of lactate dehydrogenase (LDH) were then measured.The level of apoptosis was analyzed by flow cytometry. The morphology of mitochondria was analyzed by immunofluorescence and laser confocal microscopy. The levels of mitochondrial reactive oxygen species (mtROS) were detected by immunofluorescence. Dual luciferase reporter gene assay was used to study the binding site of miR-878 and Pim1. RNA immunoprecipitation (RIP) assay was used to verify the binding relationship between miR-878 and Pim1. The gene expression levels were detected by real-time fluorescent quantitative PCR (RT-qPCR) and Western blot.Results The study found that compared with the control group, the expression of miR-878 in H/R-treated H9c2 cells was significantly increased ((1.00±0.25) vs (9.70±2.63), P<0.01). In H/R-induced cells, transfection of miR-878 inhibitor significantly increased cell viability ((46.67±3.00) vs (74.62±4.08), P<0.000 1), and decreased LDH release ((358.58±41.71) vs (179.09±15.59), P<0.000 1) and cell apoptosis rate ((43.41±0.72) vs (27.42±4.48), P<0.01). At the same time, downregulation of miR-878 expression significantly inhibited DRP1-mediated mitochondrial overdivision and mtROS production ((6.60±0.57) vs (4.32±0.91), P<0.000 1). The mechanism study showed that miR-878 could target and bind Pim1 and inhibit the expression level of Pim1 ((1.00±0.13) vs (0.38±0.03), P<0.01). Rescue experiments confirmed that down-regulation of Pim1 expression significantly reversed the anti-injury effect of miR-878 inhibitor in H9c2 cells (P<0.01), promoted mitochondrial overdivision and mtROS production ((1.00±0.12) vs (2.41±0.12), P<0.01), and decreased the expression level of p-DRP1 ((1.00±0.15) vs (0.59±0.06), P<0.05).Conclusion The present study demonstrates that miR-878 expression is upregulated in H9c2 cardiomyocytes subjected to H/R injury. Inhibition of miR-878 expression alleviates H/R-induced cardiomyocyte damage. Notably, downregulation of miR-878 significantly inhibits DRP1-mediated mitochondrial fission and mitigates mtROS production. Mechanistically, miR-878 targets and binds to the 3"-UTR of the Pim1 gene, thereby suppressing Pim1 protein expression. Collectively, these findings suggest that under H/R conditions, miR-878 promotes excessive mitochondrial fragmentation through DRP1 activation by targeting Pim1, ultimately contributing to cardiomyocyte injury. Modulation of the miR-878/Pim1 axis may represent a potential therapeutic strategy for mitigating MI/R-induced cardiac damage.]]> 2024/4/19 9:41:52 HU Shu-Wen,ZHANG Jing-Jing,BAI Ming and NIU Xiao-Wei 109true Objective To study the effects of BMI1 on the proliferation and drug resistance of cervical cancer (CC) and endometrial cancer (EC) cells. In addition, the mechanism of paclitaxel (PTX) resistance induced by BMI1 was explored.Methods In this study, we utilized the GTEx, Cbioportal, TCGA, and CPTAC databases to comprehensively analyze the mutation rate as well as mRNA and protein expression profiles of BMI1 in CC and EC. Subsequently, immunohistochemistry (IHC) analysis was employed to evaluate the protein expression levels of BMI1 in 40 pairs of CC and 40 pairs of EC tissue samples. Western blot was conducted to investigate alterations in downstream factor protein levels upon BMI1 knockdown in CC and EC cells. Furthermore, functional experiments were performed to elucidate the role of BMI1 in CC and EC cells. Finally, we assessed the synergistic anti-growth effect by combining BMI1 knockdown with paclitaxel treatment in vitro.Results The Cbioportal database revealed that BMI1 amplification, misinterpretation, and splicing occurred in 1.5% of CC patients and 1.9% of EC patients. Mining the data from TCGA and CPTAC databases, high mRNA levels of BMI1 were associated with the pathological type of CC and lower overall survival, and high protein levels of BMI1 were related to EC’s pathological type and tumor grade. Furthermore, the BMI1 protein level is overexpressed in cancer tissues of CC and EC compared with normal tissues, as detected by IHC analysis. Besides, drug sensitivity experiments showed that overexpression of BMI1 resulted in decreased sensitivity of HeLa and HEC-1-A cells to a variety of anticancer drugs, including paclitaxel. In order to further analyze the relationship between BMI1 and paclitaxel resistance, Western blot was used to detect the changes in the protein levels of downstream factors of BMI1 in HeLa and HEC-1-A cells after BMI1 knockdown. The results showed that the level of anti-apoptotic factor Bcl-2 protein decreased, while that of pro-apoptotic factor BAX increased with BMI1 knockdown. Additionally, we showed that high expression of BMI1 promoted the proliferation and migration of CC and EC cells in vitro. Moreover, CC and EC cells with low BMI1 expression were more sensitive to the paclitaxel.Conclusion The expression of BMI1 is significantly upregulated in tumor tissues from patients with cervical and endometrial cancer, and silencing BMI1 makes CC and EC cells more sensitive to paclitaxel via enhancing pro-apoptotic regulation.]]> 2024/4/19 0:00:00 ZHAO Yi-Ting,LIN Yan,YANG Wei-Li and CHEN Jun 108true in vivo, which plays an important role in apoptosis, and there is now a large amount of literature confirming that LRP1 activates the PI3K/AKT pathway, which has an anti-apoptotic effect. So can PCSK9 affect the PI3K/AKT pathway through LRP1 and thus regulate neuronal apoptosis? This deserves further investigation.The aim of this study was to explore the role of PCSK9 in mediating ox-LDL pro-apoptotic neuronal cell death and its mechanism, and then further elaborate the mechanism of hyperlipidemia leading to neurodegenerative diseases such as AD.Methods Firstly, PC12 cells were treated with different concentrations of ox-LDL (0, 25, 50, 75 and 100 mg/L) for 24 h. Oil red O staining was used to detect lipid accumulation in PC12 cells, Hoechst33258 staining and flow cytometry to detect apoptosis in PC12 cells, ELISA to detect the content of Aβ secreted by PC12, Western blot to detect expression of SREBP2, PCSK9 and LRP1. Then PC12 cells were treated with 75 mg/L ox-LDL for different times (0, 6, 12, 24, 48 h), and Western blot were performed to detect the expression of SREBP2, PCSK9 and LRP1. Finally, after transfecting 100 nmol/L PCSK9 siRNA into PC12 cells for 48 h, PC12 cells were treated with 75 mg/L ox-LDL for 24 h, Hoechst33258 staining and flow cytometry to detect apoptosis rate of PC12 cells, and Western blot to detect PCSK9, LRP1, PI3K, AKT, P-PI3K , P-AKT, NF-κB, Bcl-2, Bax, Caspase-9 and Caspase-3 expression, and ELISA detected Aβ content secreted by PC12 cells.Results ox-LDL increased lipid accumulation and promoted apoptosis and Aβ secretion in PC12 cells, as well as increasing the expression of SREBP2 and PCSK9 and decreasing the expression of LRP1 in PC12 cells. pCsk9 siRNA could be inhibited through the PI3K/AKT pathway and the NF-κB-Bcl-2/Bax-Caspase-9/3 pathway to inhibit ox-LDL-induced apoptosis in PC12 cells while increasing Aβ secretion in PC12 cells.Conclusion ox-LDL plays a bidirectional regulatory role in ox-LDL-induced apoptosis of PC12 cells by inducing an increase in PCSK9 expression and a decrease in LRP1 expression in PC12 cells, which in turn affects different signaling pathways downstream.]]> 2024/4/19 9:42:36 HE Nai-Qi,ZHAO Xue-Shan,XU Qian,ZHANG Hua-Yu,REN Zhong,TANG Zhi-Han,XIANG Qiong and LIU Lu-Shan 107true Objective This study aimed to observe the impact of sinomenine hydrochloride on the proliferation of fibroblasts and the mRNA expression of related genes in knee joint adhesion and contracture in rabbits. Additionally, we sought to explore its potential mechanisms in combating knee joint adhesion and contracture.Methods Fibroblasts were cultured in vitro, and experimental groups with varying concentrations of sinomenine hydrochloride were established alongside a control group. Cell proliferation was assessed using the CCK-8 assay. Changes in the mRNA expression of fibroblast-related genes following sinomenine hydrochloride treatment were evaluated using RT-qPCR. The impact of the drug on serum levels of inflammatory cytokines was determined using the ELISA method, and the expression of related proteins was assessed using Western blot.Results Sinomenine hydrochloride was found to inhibit fibroblast viability, with viability decreasing as the concentration of sinomenine hydrochloride increased. The effects of sinomenine hydrochloride in all experimental groups were highly significant (P<0.05). At the mRNA expression level, compared to the control group, sinomenine hydrochloride led to a significant downregulation of inflammatory cytokines in all groups (P<0.05). Additionally, the expression levels of apoptosis-related proteins significantly increased, while Bcl-2 mRNA expression decreased (P<0.05). The mRNA expression levels of the PI3K/mTOR/AKT3 signaling pathway also decreased (P<0.05). At the protein expression level, in comparison to the control group, the levels of inflammatory cytokines IL-6, IL-8, IL-1β, and TGF-β were significantly downregulated in the middle and high-dose sinomenine hydrochloride groups (P<0.05). The expression levels of cleaved-PARP, cleaved caspase-3/7, and Bax increased and were positively correlated with the dose, while the expression levels of the anti-apoptotic protein Bcl-2 and the PI3K/AKT3/mTOR signaling pathway were negatively correlated with the dose. Sinomenine hydrochloride exhibited a significant inhibitory effect on the viability of rabbit knee joint fibroblasts, which may be associated with the downregulation of inflammatory cytokines IL-6, IL-8, and IL-1β, promotion of apoptosis-related proteins cleaved-PARP, cleaved caspase-3/7, and Bax, suppression of Bcl-2 expression, and inhibition of gene expression in the downstream PI3K/AKT3/mTOR signaling pathway.Conclusion Sinomenine hydrochloride can inhibit the inflammatory response of fibroblasts in adhesive knee joints and accelerate fibroblast apoptosis. This mechanism may offer a novel approach to improving and treating knee joint adhesion.]]> 2024/4/19 9:42:39 HOU Xin-Ju,LEI Hong-Feng,CHEN Yong,LI Fu-Xi,SUN Jing-Ning,LIU Jia-Ming and MA Hong-Mei 106true Objective It was reported that the transthyretin (TTR) has a neuroprotective effect on Alzheimer’s disease (AD), which is manifested by the ability of TTR to inhibit the pathological aggregation of amyloid beta protein (Aβ). In this work, we investigated the mechanism of the interactions between TTR and Aβ at the molecular level to reveal the neuroprotective effect of TTR on AD.Methods Protein-protein docking was used to explore the models of interaction between different structural forms of TTR and Aβ, and molecular dynamics simulation was further applied to investigate the dynamic process of the interaction between the two.Results Both TTR tetramer and monomer can interact with Aβ monomer, and the thyroxine-binding channel of TTR tetramer is the main binding site of Aβ monomer. In addition, the EF helix and EF loop of TTR tetramer were also able to bind Aβ monomer. When the TTR tetramer dissociates, the hydrophobic site of the internal TTR monomer is exposed, which has a strong affinity for Aβ monomer. For the interaction between Aβ aggregates and TTR, a higher degree of aggregation can be formed between TTR monomer and Aβ aggregates due to the β-sheet-rich property of TTR monomer and Aβ aggregates, which may therefore reduce the cytotoxicity of Aβ aggregates.Conclusion Both TTR tetramer and monomer can inhibit Aβ aggregation by “sequestering” Aβ monomer, while TTR monomer can reduce the cytotoxicity of Aβ aggregates by forming large co-aggregation with Aβ aggregates. This work can provide an important theoretical basis for the design and discovery of anti-AD drugs based on the neuroprotective effects of TTR.]]> 2024/3/20 17:39:39 ZHOU Shuang-Yan,HUANG Yao-Xin,LI Xin,BAI Jia-Hui and YUAN Shuai 105true Objective Recent successful restoration of the native conformation and function of the complementary-determining regions (CDRs) of antibodies on gold nanoparticles (AuNPs) demonstrates that the era of molecular conformational engineering is dawning. Basically, molecular conformational engineering aims to precisely tune flexible non-functional molecules into special conformations to carry out novel functions, in the same way as protein folding. In order to explore the general applicability of molecular conformational engineering, as well as to reveal the mechanism of protein structure-function relationship, the objective of this work is to restore the native conformation and function of the CDRs of an antibody on platinum nanoparticles (PtNPs).Methods The CDR fragment of the anti-lysozyme antibody cAB-lys3, which has no stable conformation or function in free state, was conjugated onto the surface of PtNPs through two Pt-S bonds. The original antigen-recognizing function of the CDR restored on PtNPs was assessed by the specific inhibition of the enzymatic activity of lysozyme by the PtNP-CDR conjugates.Results After optimization of the peptide density on the surface of PtNPs and modification of PtNPs with polyethylene glycol (PEG), the resulted PtNP-based hybrid artificial antibody (PtNP-10PEG-30P1), dubbed Platinumbody, could bind specifically to lysozyme and significantly inhibit the activity of lysozyme.Conclusion This is the first time that the fragment of a protein could refold on PtNPs. Together with the previous Goldbody and Silverbody, current work demonstrates that artificial proteins could be generally created by restoration of the native conformation of natural proteins fragments on NPs.]]> 2024/3/20 17:39:44 SHENG Ling-Jie,XU Jia,DAI Jing-Jing,WANG Hai-Fang and CAO Ao-Neng 104true Objective Gastric cancer (GC) seriously affects human health and life, and research has shown that it is closely related to the serine/glycine metabolism. The proliferation ability of tumor cells is greatly influenced by the metabolism of serine and glycine. The aim of this study was to investigate the molecular mechanism of serine/glycine metabolism can affect the proliferation of gastric cancer cells.Methods In this work, a stable metabolic dynamic model of gastric cancer cells was established via a large-scale metabolic network dynamic modeling method in terms of a potential landscape description of stochastic and non-gradient systems. Based on the regulation of the model, a quantitative analysis was conducted to investigate the dynamic mechanism of serine/glycine metabolism affecting the proliferation of gastric cancer cells. We introduced random noise to the kinetic equations of the general metabolic network, and applied stochastic kinetic decomposition to obtain the Lyapunov function of the metabolic network parameter space. A stable metabolic network was achieved by further reducing the change in the Lyapunov function tied to the stochastic fluctuations.Results Despite the unavailability of a large number of dynamic parameters, we were able to successfully construct a dynamic model for the metabolic network in gastric cancer cells. When extracellular serine is available, the model preferentially consumes serine. In addition, when the conversion rate of glycine to serine increases, the model significantly upregulates the steady-state fluxes of S-adenosylmethionine (SAM) and S-adenosyl homocysteine (SAH).Conclusion In this paper, we provide evidence supporting the preferential uptake of serine by gastric cancer cells and the important role of serine/glycine conversion rate in SAM generation, which may affect the proliferation ability of gastric cancer cells by regulating the cellular methylation process. This provides a new idea and direction for targeted cancer therapy based on serine/glycine metabolism.]]> 2024/3/20 17:39:51 FAN Jun-Wu,ZHU Xiao-Mei,FAN Zhi-Yuan,LIU Bing-Ya,AO Ping and CHEN Yong-Cong 103true Objective Disruption of epithelial layer may instantaneously induce the generation of endogenous electric fields, which was proved to play an important role in guiding the cell migration and promoting wound healing. PIEZO1 is a kind of mechanic sensitive channel, may be regulated by voltage, is proved to involve in chemotactic migration of cells and play an important role in the process of wound healing. In this paper, the role of PIEZO1 and its downstream proteins FAK and integrin β1 in the electric field guided cell migration were investigated by HaCaT cells (human immortalized keratinocyte).Methods Cell migration was tracked by Living Cell Imaging System in directed current (DC) electric field (EF). Inhibitors and RNAi techniques were applied to study the function of PIEZO1 and other related proteins in electric fields. Western blot was used to detect the expression and phosphorylation levels of integrin β1 and FAK in electric field guided migration under EF stimulation.Results Piezo1 RNAi as well as Ruthenium red and GsMTx4 treatment all significantly inhibited the electrotaxis of HaCaT cells. Electric field stimulation with GsMTx4 treatment alone increased FAK phosphorylation level and the expression of integrin β1. Electric field promoted the expression level of integrin β1 and the phosphorylation level of FAK. Inhibiting the expression of PIEZO1 by RNAi significantly attenuated the phosphorylation level of FAK under EF stimulation. Inhibition of integrin β1 and FAK by inhibitor significantly decrease the electric field guided cell migration.Conclusion PIEZO1 as well as integrin β1 and FAK are involved in the electric field guided cell migration of HaCaT cells. Electric field signals regulate the expression of integrin β1 and the activation of FAK through PIEZO1-mediated signal pathway to orchestrate cell migration.]]> 2024/3/20 17:39:53 ZHANG Gui-Cheng,GONG Peng,WANG Yi-Fan and ZHAO San-Jun 102true Objective Phosphatidylinositol 3 kinases (PI3Ks) play an important role in cell directional movement by regulating F-actin. However, the structure and function of PI3Ks are complex. The role of PI3Ks in cell electrotaxis is not fully understood. Therefore, in this study, the model organism Dictyostelium discoideum cells were used as experimental materials to explore the role of PI3K1 and PI3K2 in electrotaxis.Methods Firstly, PI3K1 coding gene pikA knockout mutant and PI3K2 coding gene pikB knockout mutant were constructed by CRISPR/Cas9 system. Secondly, two mutants were placed in a DC electric field with a strength of 12 V/cm and the electrotaxis were analyzed.Results Data analysis showed that the direction index of wild-type cells in DC electric field was (0.86±0.03), while the direction index of pikA- and pikB- mutants in DC electric field was (0.95±0.02) and (0.94±0.03), respectively. In addition, the average trajectory speed of wild-type cells in the electric field was (3.34±0.08) μm/min, while the average trajectory speed of pikA- and pikB- mutants were (4.85±0.20) μm/min and (5.48±0.15) μm/min, respectively. The t test showed that there were significant differences in the directedness index and speed between the mutant and wild type. Western blot results showed that both phosphorylated Akt and phosphorylated ERK were significantly increased in pikA- and pikB- mutants.Conclusion PI3K1 and PI3K2 may inhibit the electrotaxis of Dictyostelium discoideum cells by increasing the activity of Akt and ERK.]]> 2024/3/20 17:39:55 GE Xiao-Xue,JIANG Rui-Da,WANG Xiao-Yan,GAO Jing and GAO Run-Chi 101true Objective At present, the matching reagents of commercially available rapid DNA instruments based on microfluidics chip technology are autosome short tandem repeat (STR) individual identification reagents. The non-recombining part of the human Y chromosome is widely used in forensic DNA analysis, particularly in cases where standard autosomal DNA profile is uninformative. Y-STR loci are useful markers to identify males and male lineages in forensic practice. In order to achieve rapid and fully integrated detection of Y-STR loci, this study constructed the RTyper Y27 microfluidic chip rapid detection system and validated the performance of this system.Methods The system was verified and evaluated by sensitivity, success rate, typing accuracy, peak height balance, sizing precision and accuracy, mock case sample tests, mixture detection ability, and inhibition tolerance.Results Complete Y-STR profiles can be obtained when the template amount of DNA standard 9948 was ≥8 ng, the number of blood cards was ≥3 pieces, and the number of oral swab scrapings was ≥7 times. The success rate of fully integrated detection was 91.52%, and the concordance rates was 99.74% for 165 testing samples. The success rate of 115 blood spots in these samples was 90.43%, with a typing accuracy of 99.65%, the success rate of 50 buccal swabs was 94%, with a typing accuracy of 99.92%. There was no significant difference in typing accuracy between blood spots and buccal swab samples. The peak height ratio between different fluorescence channels was 89.81%. The standard deviation of allelic ladder for 10 runs was within 0.5 bp. The size differences between allele and corresponding allele in allelic ladder was within 0.5 bp. The maximum precision CV values within and between batches were 0.48% and 0.68%, respectively, which were lower than 15%. These data indicate that the system has good accuracy and precision. The system was capable of accurately typing oral swabs, blood cards, saliva cards, cigarette butts, blood swabs and seminal stains. Complete Y-STR profiles can be obtained and distinguish at the 1∶3 ratio of minor and major contributors in artificial male DNA mixtures. Complete Y-STR genotyping can be obtained under the interference of inhibitors, such as different concentrations of humic acid (50-400 mg/L), indigotin (20-100 nmol/L) and hemoglobin (100-500 μmol/L).Conclusion In this study, the RTyper Y27 microfluidic chip rapid amplification system is combined with the Quick TargSeq 1.0 integrated system, and the Y-STR profile can be obtained in approximately 2 h. Through a series of verification experiments, the results show that the system has good repeatability, accuracy and stability, can meet the on-site Y-STR detection requirements, and can be used in forensic practice.]]> 2024/3/20 17:39:58 WANG Dao-Yu,WAN Qun,ZHUANG Bin,ZHAO Li-Jian,HAN Jun-Ping and LI Cai-Xia 100true Objective The traditional Chinese medicine Strychnos nux-vomica L. (SN) has the clinical effect of reducing swelling and relieving pain; however, SN is toxic due to its alkaloid components. Little is known about the endogenous metabolic changes induced by SN toxicity in rats and their potential effects on the metabolic dysregulation of intestinal microbiota. Therefore, toxicological investigation of SN is of great significance to its safety assessment. In this study, the toxic mechanisms of SN were explored using a combination of metabonomics and 16S rRNA gene sequencing.Methods The toxic dose, intensity, and target organ of SN were determined in rats using acute, cumulative, and subacute toxicity tests. UHPLC-MS was used to analyze the serum, liver, and renal samples of rats after intragastric SN administration. The decision tree and K Nearest Neighbor (KNN) model were established based on the bootstrap aggregation (bagging) algorithm to classify the omics data. After samples were extracted from rat feces, the high-throughput sequencing platform was used to analyze the 16S rRNA V3-V4 region of bacteria.Results The bagging algorithm improved the accuracy of sample classification. Twelve biomarkers were identified, where their metabolic dysregulation may be responsible for SN toxicity in vivo. Several types of bacteria such as Bacteroidetes, Anaerostipes, Oscillospira and Bilophila, were demonstrated to be closely related to physiological indices of renal and liver function, indicating that SN-induced liver and kidney damage may be related to the disturbance of these intestinal bacteria.Conclusion The toxicity mechanism of SN was revealed in vivo, which provides a scientific basis for the safe and rational clinical use of SN.]]> 2024/2/22 16:33:38 WANG Xi-Ye,BAO Le-Er,JIANG Ming-Yang,LI Dan and BAI Mei-Rong 99true Objective To investigate the genetic polymorphism and structure of 47 autosomal microhaplotypes in the Han population in Changshu City, Jiangsu Province, and to evaluate the forensic efficiencies and forensic parameters.Methods The DNA library of unrelated individual samples was prepared according to MHSeqTyper47 kit manual and sequenced on the MiSeq FGx platform. Microhaplotype genotyping and sequencing depth statistics were processed using MHTyper. The genetic information of samples was then evaluated. The fixation index and genetic distance between the Jiangsu Changshu population and the reference populations in the 1000 Genomes Project phase 3 (1KG) were calculated, and forensic parameters were evaluated.Results The fixation index and genetic distance between the Han population in Changshu, Jiangsu, and the CHB (Han Chinese in Beijing, China) reference population in 1KG were the lowest. The effective allele number (Ae) of each locus is also the closest between the two populations. The combined matching probability (CMP) of the Changshu Han population is close to the 5 populations of the East Asian reference super-population in 1KG, which is 1.25×10-36, and the combined probability of exclusion reached 0.999 999 999 964 1.Conclusion This study reported the genetic polymorphism and allele frequency of 47 microhaplotypes in a Han population in Changshu City, Jiangsu Province. This information provides a data basis for 47 microhaplotypes in forensic applications. In addition, the polymorphism differences between the 1KG reference population and the Han population in Changshu, Jiangsu were compared, and the genetic structure of 47 microhaplotypes in the Han population in Changshu, Jiangsu was revealed. In general, the reference data of the East Asian super-population in 1KG is more in line with the genetic characteristics of Han population in Changshu, Jiangsu.]]> 2024/2/22 16:33:40 PAN Kun-Peng,FENG Yao-Sen,YU Wen-Shuai,LIU Zong-Wei,YAO Yi-Ren,ZHAO Jie,KANG Ke-Lai,ZHANG Chi,WANG Le and WU Jian 98true Objective A growing body of research suggests a strong link between metabolic imbalance and Alzheimer’s disease (AD). In recent years, the development of metabolomics makes it possible to study the characteristic changes of peripheral metabolism in AD. Serum levels of tryptophan and tyrosine were associated with mild cognitive impairment (MCI) and AD. The purpose of this study is to further characterize tryptophan and tyrosine levels in MCI and AD.Methods We stratified 765 participants from the Alzheimer’s Disease Neuroimaging Initiative-1 (ADNI-1) cohort into cognitively normal (CN; n=207), stable mild cognitive impairment (sMCI; n=201), progressive mild cognitive impairment (pMCI; n=171), and dementia due to AD (n=186). Serum tryptophan and tyrosine were analyzed for diagnostic value of MCI and AD. To evaluate the relationships between serum tryptophan and tyrosine and cerebrospinal fluid (CSF) biomarkers, brain structure (magnetic resonance imaging, MRI), cerebral glucose metabolism (18F-fluorodeoxyglucose-positron emission tomography, FDG-PET), and cognitive declines, through different cognitive subgroups.Results Serum tryptophan was decreased in patients with AD, pMCI or sMCI compared with CN. Serum tyrosine was decreased in patients with AD or pMCI compared with CN. Serum tryptophan has diagnostic value for pMCI and AD. Serum tyrosine has diagnostic value for AD.Conclusion Serum tryptophan and tyrosine contribute to the early diagnosis of AD. The detection of tryptophan and tyrosine can contribute to the pathogenesis of AD. Serum tryptophan and tyrosine were not significantly associated with core AD markers, cognitive function, brain structure and brain metabolism, so serum tryptophan and tyrosine may not be good peripheral AD biomarkers.]]> 2024/2/22 16:33:41 REN Shu-Jiang,JING Xiao-Jun and ZHANG Hua 97true Objective Direct continuous monitoring of arterial blood pressure is invasive and continuous monitoring cannot be achieved by traditional cuffed indirect blood pressure measurement methods. Previously, continuous non-invasive arterial blood pressure monitoring was achieved by using photoplethysmography (PPG), but it is discrete values of systolic and diastolic blood pressures rather than continuous values constructing arterial blood pressure waves. This study aimed to reconstruct arterial blood pressure wave signal based on CNN-LSTM using PPG to achieve continuous non-invasive arterial blood pressure monitoring.Methods A CNN-LSTM hybrid neural network model was constructed, and the PPG and arterial blood pressure wave synchronized recorded signal data from the Medical Information Mart for Intensive Care (MIMIC) were used. The PPG signals were input to this model after noise reduction, normalization, and sliding window segmentation. The corresponding arterial blood pressure waves were reconstructed from PPG by using the CNN-LSTM hybrid model.Results When using the CNN-LSTM neural network with a window length of 312, the error between the reconstructed arterial blood pressure values and the actual arterial blood pressure values was minimal: the values of mean absolute error (MAE) and root mean square error (RMSE) were 2.79 mmHg and 4.24 mmHg, respectively, and the cosine similarity is the optimal. The reconstructed arterial blood pressure values were highly correlated with the actual arterial blood pressure values, which met the Association for the Advancement of Medical Instrumentation (AAMI) standards.Conclusion CNN-LSTM hybrid neural network can reconstruct arterial blood pressure wave signal using PPG to achieve continuous non-invasive arterial blood pressure monitoring.]]> 2024/2/22 16:33:43 WU Jia-Ze,LIANG Hao and CHEN Ming 96true Objective TePixD (Tll0078) is a blue light-using flavin (BLUF) photoreceptor protein from Thermosynechococcus elongatus BP-1. TePixD protein has a conserved Tyr8-Gln50-Met93 triad around the FAD pocket to mediate the proton-coupled electron transfer (PCET) process. But the detailed light response mechanism needs further study. We aimed to elucidate the structure and biochemical properties of TePixD mutants at key light response sites to analyze the light response process of TePixD.Methods We employed X-ray crystallography to resolve the crystal structure of the TePixD Y8F mutant. The side chain of Tyr8 is involved in PCET while Phe8 in mutation loses the function due to the loss of its hydroxyl group. We compared the structure of TePixD Y8F mutation to TePixD wild type (WT) and its homology protein SyPixD Y8F. Using multi-angle light scattering (MALS), we analyzed the oligomerization of multiple TePixD mutations (Y8F, Q50L, W91F, Y8F/W91F, and Q50L/W91F), focusing specifically on mutational sites that are critical residues for the protein’s photo response to dark and light conditions.Results We resolved the crystal structure of TePixD Y8F mutant at a resolution of 2.54 ? and found that it shares a similar overall structure with the TePixD WT but exhibits significant differences from the SyPixD Y8F structure. Biochemical analysis revealed differences in molecular mass and elution profiles between the TePixD mutants and the WT under dark and light conditions, indicating the perturbation on the light-induced conformational change by the mutants.Conclusion Our structure determination and biochemical analyses will add information to reveal the light response mechanism of BLUF proteins.]]> 2024/2/22 16:33:46 HU Rui-Xing,ZHOU Ya-Lin,LIN Lin,DING Bei and LU Qing 95true Objective Existing artificial vision devices can be divided into two types: implanted devices and extracorporeal devices, both of which have some disadvantages. The former requires surgical implantation, which may lead to irreversible trauma, while the latter has some defects such as relatively simple instructions, limited application scenarios and relying too much on the judgment of artificial intelligence (AI) to provide enough security. Here we propose a system that has voice interaction and can convert surrounding environment information into tactile commands on head and neck. Compared with existing extracorporeal devices, our device can provide a larger capacity of information and has advantages such as lower cost, lower risk, suitable for a variety of life and work scenarios.Methods With the latest remote wireless communication and chip technologies, microelectronic devices, cameras and sensors worn by the user, as well as the huge database and computing power in the cloud, the backend staff can get a full insight into the scenario, environmental parameters and status of the user remotely (for example, across the city) in real time. In the meanwhile, by comparing the cloud database and in-memory database and with the help of AI-assisted recognition and manual analysis, they can quickly develop the most reasonable action plan and send instructions to the user. In addition, the backend staff can provide humanistic care and emotional sustenance through voice dialogs.Results This study originally proposes the concept of “remote virtual companion” and demonstrates the related hardware and software as well as test results. The system can not only achieve basic guide functions, for example, helping a person with visual impairment to shop in supermarkets, find seats at cafes, walk on the streets, construct complex puzzles, and play cards, but also can meet the demand for fast-paced daily tasks such as cycling.Conclusion Experimental results show that this “remote virtual companion” is applicable for various scenarios and demands. It can help blind people with their travels, shopping and entertainment, or accompany the elderlies with their trips, wilderness explorations, and travels.]]> 2024/1/19 11:50:48 GE Song,HUANG Xuan-Tuo,LIN Yan-Ni,LI Yan-Cheng,DONG Wen-Tian,DANG Wei-Min,XU Jing-Jing,YI Ming and XU Sheng-Yong 94true Objective The scale of microalgae farming industry is huge. During farming, it is easy for microalgae to be affected by miscellaneous bacteria and other contaminants. Because of that, periodic test is necessary to ensure the growth of microalgae. Present microscopy imaging and spectral analysis methods have higher requirements for experiment personnel, equipment and sites, for which it is unable to achieve real-time portable detection. For the purpose of real-time portable microalgae detection, a real-time microalgae detection system of low detection requirement and fast detection speed is needed.Methods This study has developed a microalgae detection system based on deep learning. A microscopy imaging device based on bright field was constructed. With imaged captured from the device, a neural network based on YOLOv3 was trained and deployed on microcomputer, thus realizing real-time portable microalgae detection. This study has also improved the feature extraction network by introducing cross-region residual connection and attention mechanism and replacing optimizer with Adam optimizer using multistage and multimethod strategy.Results With cross-region residual connection, the mAP value reached 0.92. Compared with manual result, the detection error was 2.47%.Conclusion The system could achieve real-time portable microalgae detection and provide relatively accurate detection result, so it can be applied to periodic test in microalgae farming.]]> 2024/1/19 11:50:58 XIANG Rui-Jie,LIU Hao,LU Zhen,XIAO Ze-Yu,LIU Hai-Peng,WANG Yin-Chu,PENG Xiao and YAN Wei 93true Objective Transcription factor NFE2 was observed abnormal expression in myeloproliferative neoplasm (MPN) patients. However, how NFE2 is transcriptionally regulated remains ambiguous. This study aims to explore the elements and molecular mechanisms involved in the transcriptional regulation of NFE2.Methods Active enhancers were predicted by public NGS data and conformed experimentally via dual luciferase reporter assay. After that, PRO-seq and GRO-seq data was used to detect enhancer RNAs transcribed from these enhancers. RACE was utilized to clone the full length enhancer RNA (eRNA) transcripts, and RT-qPCR was used to measure their expression in different leukemia cell lines as well as the transcript levels during induced differentiation. Finally, to investigate the molecular function of the eRNA, overexpression and knockdown of the eRNA via lentivirus system was performed in K562 cells.Results We identified three enhancers regulating NFE2 transcription, which located at -3.6k, -6.2k and +6.3k from NFE2 transcription start site (TSS) respectively. At the -3.6k enhancer, we cloned an eRNA transcript and characterized that as a lncRNA which was expressed and located in the nucleus in three types of leukemia cell lines. When this lncRNA was overexpressed, expression of NFE2 was upregulated and decreases of K562 cell proliferation and migration ability were observed. While knocking down of this lncRNA, the level of NFE2 decreases correspondingly and the proliferation ability of K562 cells increases accordingly.Conclusion We identified an enhancer lncRNA that regulates NFE2 transcription positively and suppresses K562 cell proliferation.]]> 2024/1/19 11:51:02 LU Yan-Fei,QU Song-Ya,ZHU Jing-Jing,LIU Chao,WANG Jian,HAN Bing-She and ZHANG Jun-Fang 92true Objective Our recent study has demonstrated that extracellular acidification promotes lipid accumulation in macrophages via the activation of acid sensing ion channel 1 (ASIC1), but the underlying mechanism remains unclear. This study aims to explore the effect of extracellular acidification on macrophage lipophagy and the underlying mechanism.Methods RAW264.7 macrophages were incubated with 25 mg/L ox-LDL in a pH 6.5 culture medium for 24 h to build macrophage-derived foam cell models induced by extracellular acidification. Then, RAW264.7 macrophages were cultured in the acidic medium of pH 6.5 with or without PcTx-1 (ASIC1 specific blocker, 10 μg/L) or Nec-1 (RIP1 specific inhibitor, 20 μmol/L) for 24 h, intracellular lipid accumulation was observed by oil red O staining. The expressions of total ASIC1, plasma membrane ASIC1, RIP1, p-RIP1 Ser166, TFEB, p-TFEB Ser142, LC3 and p62 were measured by Western blot. The co-localization of lipids (indicated by Bodipy) with LC3II (autophagosomes) and LAMP1 (lysosomes) was analyzed by a confocal laser scanning microscopy, respectively. Morphological changes of lipophagy in the cells were observed by using transmission electron microscopy. ABCA1-mediated cholesterol efflux was determined by cholesterol fluorescence kits.Results Compared with pH 7.4+ox-LDL group, the intracellular lipid accumulation in the pH 6.5+ox-LDL group was significantly increased. Meanwhile, the expressions of plasma membrane ASIC1, p-RIP1 Ser166, p-TFEB Ser142, and p62 proteins were elevated significantly, while LC3II protein level and LC3II/LC3I ratio were decreased. Accordingly, compared with pH 7.4+ox-LDL group, the macrophage lipophagy of the pH 6.5+ox-LDL group was inhibited as indicated by the decreased localization of lipid droplets with LC3 and LAMP1, a decrease in the number of lipophagosomes as well as an increase in lipid droplets. Furthermore, ATP binding cassette transporter A1 (ABCA1)-dependent cholesterol efflux from the macrophages of pH 6.5+ox-LDL group reduced dramatically. However, these above effects of extracellular acidification on RAW264.7 macrophages were abolished by PcTx-1 and Nec-1, respectively.Conclusion These findings suggest extracellular acidification promotes the phosphorylation of TFEB at Ser142 via activating ASIC1/RIP1 pathway, thereby impeding lipophagy in RAW 264.7 macrophages, and that ASIC1 may be a new potential target for preventing aberrant lipid accumulation diseases including atherosclerosis.]]> 2024/1/19 11:51:06 LIU Juan,OU Xiang,LIU Qing,GUO Miao,NING Zi-Ping,GU Hong-Feng and TANG Ya-Ling 91true Objective Although expression of the TEAD1 protein in preadipocytes has been established, its function remains unclear. In this study, we sought to detect transcripts of TEAD1 in chicken and to examine the effects of this protein on the proliferation, migration, apoptosis, and differentiation of immortalized chicken preadipocyte cell lines (ICP1).Methods The full-length sequence of the TEAD1 gene was cloned and the two transcripts were subjected to bioinformatics analysis. The subcellsular localization of TEAD1 transcripts was determined based on indirect immunofluorescence. The effects of TEAD1 transcripts overexpression on the proliferation of ICP1 cells were examined by RT-qPCR, CCK-8, and EdU assays; the effects of TEAD1 transcripts on ICP1 cells migration were examined based on the scratch test; and the effects of TEAD1 transcripts overexpression on ICP1 cells apoptosis were analyzed using apoptosis-Hoechst staining and RT-qPCR. The expression of TEAD1 transcripts in different tissues, cells lines, and ICP1 at different periods of differentiation was analyzed by RT-qPCR. The effects of TEAD1 transcripts overexpression on lipid droplet accumulation and adipogenic-related gene expression in ICP1 cells were analyzed based on Oil Red O and BODIPY staining, RT-qPCR, Western blot, and dual-luciferase reporter gene assays. Finally, the content of triglyceride (TG) was measured in TEAD1 overexpressed ICP1 cells.Results The full-length TEAD1 was cloned and two TEAD1 transcripts were identified. The TEAD1-V1 protein was found to be localized primarily in the cell nucleus, whereas the TEAD1-V2 protein is localized in the cell cytoplasm and nucleus. The overexpression of both TEAD1-V1 and TEAD1-V2 significantly inhibited the proliferation of ICP1 cells. Whereas the overexpression of TEAD1-V1 promoted ICP1 cell migration, the overexpression of TEAD1-V2 had no significant effects on ICP1 migration; the overexpression of both TEAD1-V1 and TEAD1-V2 significantly promoted the apoptosis of ICP1 cells. We found that the different transcripts of TEAD1 have similar expression pattern in different tissues and cells lines. During induced preadipocyte differentiation, the expression of these genes initially declined, although subsequently increased. Overexpression of TEAD1-V1 promoted a significant reduction in lipid droplet formation and inhibited C/EBPα expression during the differentiation of ICP1 cells (P<0.05). However, the overexpression of TEAD1-V2 had no significant effect on lipid droplet accumulation or the expression of adipogenic-related proteins (P>0.05). Overexpression of TEAD1-V1 significantly decreased triglyceride content in ICP1 cells (P<0.05), while overexpression of TEAD1-V2 had no effect on triglyceride content in ICP1 cells (P>0.05).Conclusion In this study, for the first time, identified two TEAD1 transcripts. Overexpressed transcripts TEAD1-V1 and TEAD1-V2 both inhibited the proliferation of chicken preadipocytes and promoted apoptosis of chicken preadipocytes. TEAD1-V1 inhibited the differentiation of preadipocytes and promoted the migration of preadipocytes, while TEAD1-V2 had no effect on the differentiation and migration of preadipocytes.]]> 2024/1/19 11:51:30 PENG Min,XU Hu,JIA Zi-Qiu,YANG Qing-Zhu,PAN Lin,WANG Wei-Yu,KONG Ling-Zhe and SUN Ying-Ning 90true Objective To investigate the expression of cyclin-dependent kinase 8 (CDK8) in esophageal squamous cell carcinoma (ESCC) and its effect on ESCC cells, and to explore its potential molecular mechanism.Methods The expression level of CDK8 mRNA was analyzed using UALCAN database, and then the expression level of CDK8 protein in tumor tissues of ESCC patients was detected by immunohistochemistry (IHC). Esophageal cancer cell lines Kyse-30 and Kyse-150 were stably transfected with lentivirus to achieve knockdown and overexpression of CDK8. EdU proliferation assay, cell colony formation assay, cell cycle assay, cell scratch assay and invasion assay were used to explore the effect of CDK8 protein expression level on the phenotype of ESCC cells. Subsequently, the effect of CDK8 on the growth of esophageal cancer xenografts in vitro was observed by subcutaneous tumor formation assay in mice. Finally, the expression of proliferation and metastasis related proteins was detected by Western blot.Results CDK8 showed high transcription and protein expression levels in ESCC tissues compared with normal esophageal tissues. Knockdown of CDK8 expression significantly inhibited the proliferation, migration and invasion of ESCC cells. In addition, inhibition of CDK8 expression significantly affected the JAK2/STAT3 pathway and the expression of E-cadherin/N-cadherin, while overexpression of CDK8 reversed these effects. Inhibition of STAT3 pathway reversed the promoting effect of CDK8 overexpression on ESCC cell phenotype.Conclusion CDK8 is a cancer-promoting factor of ESCC, which mediates the phosphorylation of JAK2/ STAT3 and epithelial-mesenchymal transition (EMT).]]> 2024/6/19 14:40:02 QU Hang-Shuai,TIAN Xiong,PAN Yi-Xiao,BAO Jia-Qian,YE Lu-Xia and ZHENG Jing-Min 89true Objective Formononetin (FOR), a traditional Chinese medicine, has been widely used for nerve protection and nerve function rehabilitation after cerebral stroke. However, the role of FOR in autophagic lysosome function in cerebral ischemia-reperfusion damage has not been investigated. This study aimed to explore whether the therapeutic benefits of FOR were influenced by the regulation of autophagy flux.Methods Male Sprague-Dawley rats were separated into sham, model, and MCAO+FOR (30 mg/kg) groups after undergoing middle cerebral artery occlusion (MCAO) and ischemia-reperfusion (I/R). Then, the brain tissues in the ischemic penumbra were obtained to detect the proteins in autophagic/lysosomal pathway with antibodies of Beclin-1, LC3, SQSTM1/P62, Ubiquitin, LAMP-2, Cathepsin B (CTSB) and Cathepsin D (CTSD) by Western blot and immunofluorescence, respectively. Meanwhile, the therapeutic effectiveness was evaluated by measuring infarct volume, neurological impairments, and neuronal necrosis.Results The findings of this study demonstrate that FOR treatment exhibits a dual effect by enhancing the autophagic activities of Beclin-1 and LC3 in neurons, while simultaneously improving the autophagic clearance function, as evidenced by reinforced lysosomal activities of LAMP-2, CTSB, and CTSD, as well as reduced autophagic accumulation of Ubiquitin and P62 in the MCAO+FOR group compared to the MCAO group. Additionally, 7 d of FOR treatment dramatically reduced neurological deficits, infarct volume, and neuronal death caused by cerebral ischemia.Conclusion These findings suggest that the neuroprotective mechanism of FOR therapy in accelerating recovery from ischemic stroke may involve the increase of autophagy flux in the penumbra.]]> 2024/12/23 14:28:31 GUO Tao,ZUO Han-Jun,SHI Jin-Sha,SHI Hao-Long,WANG Zhao,CHEN Bo-Lin and Li Juan-Juan 88true Objective Chronic stress can induce cognitive dysfunction, but the underlying mechanisms remain unknown. Studies have confirmed that the high mobility group box 1/Toll-like receptor 4 (HMGB1/TLR4) pathway is closely associated with cognitive impairment. Therefore, this research aimed to explore whether the HMGB1/TLR4 pathway involves in chronic stress-induced cognitive dysfunction.Methods The chronic unpredictable mild stress (CUMS) mouse model was established by randomly giving different types of stress every day for four consecutive weeks. Cognitive function was detected by novel object recognition test, Y-maze test, and Morris water maze test. The protein expressions of HMGB1, TLR4, B-cell lymphoma 2 (BCL2), and BCL2 associated X (BAX) were determined by Western blot. The damage of neurons in the hippocampal CA1 region was observed by hematoxylin-eosin (HE) staining.Results The protein expressions of HMGB1 and TLR4 were significantly increased in the hippocampus of chronic stress mice. Furthermore, inhibition of the HMGB1/TLR4 pathway induced by ethyl pyruvate (EP, a specific inhibitor of HMGB1) and TAK242 (a selective inhibitor of TLR4) treatment attenuated cognitive impairment in chronic stress mice, according to the novel object recognition test, Y-maze test, and Morris water maze test. In addition, administration of EP and TAK242 also mitigated the increase of apoptosis in the hippocampus of chronic stress mice.Conclusion These results indicate that the hippocampal HMGB1/TLR4 pathway contributes to chronic stress-induced apoptosis and cognitive dysfunction.]]> 2024/6/7 19:25:59 HU Wen,KUANG Xin,FENG Xin-Xiang,ZHONG Wen-Long,JIN Xin,JIANG Jia-Mei and ZOU Wei 87true Objective This work examines the impact of external electric fields at terahertz (THz) frequencies on double-stranded deoxyribonucleic acid (dsDNA) systems adsorbed on Au(111) surfaces in aqueous environments.Methods The investigation utilizes a molecular dynamics (MD) approach at the atomic level and vibrational dynamics calculations using the GolDNA-Amber force field.Results The results reveal that the sugar-phosphate backbone of the DNA exhibits reduced adherence to the gold surface, while the side chains display a stronger affinity. When subjecting the hydrated DNA strands to an electric field with frequencies up to 10 THz, peak intensities of vibrational dynamic density (VDoS) are observed at five different frequencies. Moreover, the strong electric field causes hydrogen bonds in the DNA within the slit to break. The sensitivity to the electric field is particularly pronounced at 8.8 THz and 9.6 THz, with different vibrational modes observed at varying electric field strengths.Conclusion These findings contribute to an enhanced understanding of the molecular organization of gold-plated charged biological interfaces.]]> 2024/6/26 0:00:00 JIANG Duan-Jie,WANG Yan-Hong,WU Jing-Zhi and ZHANG Rui 86true Objective The inference of biogeographical ancestry (BGA) using DNA is a significant focus within anthropology and forensic science. Current methods often utilize dozens of ancestry-informative SNPs, employing principal component analysis (PCA) and likelihood ratios (LR) to ascertain individual ancestries. Nonetheless, the selection of these SNPs tends to be population-specific and shows limitations in population differentiation. With the development of high-throughput sequencing technologies, acquiring high-density SNP datasets has become easier, challenging traditional statistical models which are often reliant on prior assumptions and struggle with high-density genetic data. The integration of machine learning, which prioritizes data learning and algorithmic iteration over prior knowledge, has propelled forward new developments in BGA research. This study aims to construct a BGA inference model suitable for high-density SNP data, characterized by broad population applicability, higher accuracy, and strong generalization capabilities.Methods Initially, intersection sites of autosomes from the phase III data of the 1000 Genomes Project and commonly used commercial chips were selected to build a reference dataset after thorough site quality control and filtering. This dataset was analyzed using PCA and ADMIXTURE to study population clustering, ancestral component mixing, and genetic substructures. Utilizing spaces of different principal component (PC), combinations, this study visually assessed the PCs’ capabilities to differentiate between continental and intercontinental populations. Following this, the study employed the supervised learning classification model XGBoost, establishing a multidimensional PC-based PCA-XGBoost model with hyperparameters set through ten-fold cross-validation and a greedy strategy. Subsequently, the model was optimized and evaluated based on the LR, considering accuracy and runtime to determine the optimal number of PCs and training rounds, culminating in the study’s optimal BGA inference model. Finally, the performance of the model was subsequently validated at national and regional levels using test sets from other public data to assess its post-optimization generalization capabilities.Results The reference dataset created contains 307 866 SNP sites. Top PCs reflect varying levels of population differentiation capabilities, with some PCs showing population specificity. Under smaller K values in ADMIXTURE results, genetic ancestral components between continents are elucidated, while larger K values reveal some specific ancestral components of certain populations within continents. The number of PCs and training rounds significantly affect the classification accuracy and efficiency of the XGBoost supervised model. With LR-based evaluation methods, the optimized PCA-XGBoost model achieved a continental prediction accuracy of over 98% in the reference set. For subcontinental population levels within the continents, the model achieved an accuracy of over 95% in the reference set and over 90% in the test set.Conclusion The reference dataset effectively represents the genetic substructures of populations at selected sites. Information derived from PC dimensions significantly aids in population differentiation and inference issues, and incorporating more PC dimensions as features in supervised learning models can increase the accuracy of BGA inference. The model of this study is suitable for high-density SNP data and is not confined to specific regional populations, offering enhanced population-wide applicability. Compared to previous ancestry inference models, the optimized PCA-XGBoost model demonstrates high intercontinental population predictive accuracy. LR-based evaluation methods further enhance the reliability of predictions. Additionally, the model’s strong generalization capabilities suggest that updating the reference population data could enable more detailed population analysis and inference.]]> 2024/6/6 0:00:00 YAO Hao-Tian,JIANG Li,WANG Chun-Nian,FAN Hong and LI Cai-Xia 85true Objective GQDs has become a superstar among zero-dimensional carbon-based materials. As one of the most abundant and important biological elements, its unique optical properties, high dispersion and biocompatibility have attracted extensive attention from scientists. This paper aims to investigate the effect of GQDs on cell viability, apoptosis and inflammatory factor expression in RAW264.7 macrophages and evaluate cell imaging capability of GQDs in vitro, which could provide theoretical basis for the safe application of GQDs in biomedical field.Methods Graphene oxide was prepared by modified Hummer’s method. H2O2 and W18O49 interacted with each other under hydrothermal conditions to produce hydroxyl radicals, which can cut graphene oxide into GQDs using a top-down approach. The microstructure of GQDs was analyzed in detail by X-ray powder diffraction, X-ray photoelectron spectroscopy, transmission electron microscopy, atomic force microscopy, scanning electron microscopy and Fourier infrared transform. The biocompatibility of GQDs on macrophage was evaluated by CCK-8 and dead/alive staining. Flow cytometry results showed the apoptosis of RAW264.7 macrophages induced by GQDs. mRNA expression of inflammatory factors was evaluated by RT-qPCR. Cell imaging is exhibited by laser scanning confocal.Results Hydroxyl radicals are produced by H2O2 and W18O49 under hydrothermal conditions, which contribute to cut graphene oxide into 3-5 nm GQDs in one step. The quantum yield of this method is 43%. Fluorescence lifetime of these blue GQDs is 1.67 ns. The Zigzag-type site and defect state of the triplet carbene radical lead to the excitation wavelength dependence of GQDs, and the optimal excitation and emission wavelengths are 330 nm and 400 nm, respectively. The boundary effect and amphiphilicity of quantum dots make GQDs possess abundant functional groups, vacancy defects and high dispersion, which results in GQDs exhibits good water solubility. RAW264.7 macrophages are incubated with different concentration in DEME medium for 24 h, 48 h and 72 h to evaluate cell. The survival rate of RAW264.7 cells is significantly dependent on the concentration and time of GQDs. CCK-8 and dead/alive staining show that GQDs have high biocompatibility. The effect of 200 mg/L GQDs on apoptosis of RAW264.7 cells is revealed by the scatter plot of bivariate flow cytometry. Under the stimulation of LPS+INF-γ, the expression of TNF-α was increased in RAW264.7 cells, which co-acted with other cytokines to participate in the immune response of RAW264.7 cells in vitro, and mediated the production of IL-1β inflammatory factor in RAW264.7 cells, thereby inducing apoptosis of RAW264.7 cells. The results of RT-qPCR showed that GQDs can inhibit the growth of RAW264.7 cells in vitro, and stimulate them to increase TNF-α expression in RAW264.7 cells, which make cell membrane rupture and produce IL-1β inflammatory factors to induce cell apoptosis. The high biocompatibility of GQDs is attributed to the rich oxygen-containing functional groups (―COOH, ―OH, and CO) on the surface of GQDs, which makes its surface negatively charged and easy to be swallowed into the cell interior when interacting with the cell membrane with low affinity. Transmission electron microscopy (TEM) observed that the GQDs were swallowed into the cells. Furthermore, laser confocal results displayed that blue GQDs has certain ability of cell imaging in vitro.Conclusion The water solubility, low toxicity, fluorescence properties and the induction effect of inflammatory factors of GQDs provide broad prospects for their application in the field of immunotherapy and cell imaging in the future.]]> 2024/11/22 0:00:00 LIU Qi,XU Hai-Yan,SU Yu-Xuan,ZHOU Kai-Hong and LI Chang-Yan 84true Objective To explore the mechanism of treadmill exercise against type 2 diabetes mellitus (T2DM) with non-alcoholic fatty liver disease (NAFLD) based on the regulator effects of exercise on ferroptosis.Methods Eight 8-week-old male m/m mice were used as control group (Con, n=8), and db/db mice of the matched age were randomly divided into T2DM model group (db/db, n=8), exercise group (db+Exe, n=8), p38 mitogen-activated protein kinase (MAPK) inhibitor group (db+SB203580, n=8) and exercise combined with p38 MAPK inhibitor group (db+Exe+SB203580, n=8). After one-week adaptive feeding, the mice in the db+Exe group and db+Exe+SB203580 group underwent moderate intensity treadmill exercise for 40 min/d, 5 d/week lasting 8 weeks. The db+SB203580 group and db+Exe+SB203580 group were treated with SB203580 (a specific inhibitor of p38 MAPK) with a dose of 5 mg/kg, 5 d/week for 8 weeks. And the exercise intervention was performed 2 h later after the intraperitoneal injection of SB203580. The body weight and fasting blood glucose of mice were measured regularly every week during the experiment. After 24 h of the last intervention, the mice were weighted, the liver tissues were taken, weighted and the liver index was calculated. The pathological changes of liver were determined by Oil Red O and hematoxylin-eosin (HE) staining. The levels of blood lipids, liver function, Fe2+ and oxidative stress markers of liver were measured by enzyme linked immunosorbent assay (ELISA). The related mRNA expression levels of lipogenesis and inflammation were evaluated by quantitative reverse transcriptase-mediated PCR (qRT-PCR). The related protein expression levels of lipogenesis and ferroptosis in liver were determined by immunohistochemical (IHC) staining and Western blot.Results The body weight, fasting blood glucose, liver index, blood lipid and transaminase levels in the db/db group were significantly increased compared with the Con group. HE and Oil Red O staining showed severe lipid accumulation and ballooning change in the liver of db/db mice. Biochemical tests showed that Fe2+ and MDA level of liver constitution homogenate increased, while GSH level decreased significantly. The results of qRT-PCR showed that the mRNA levels of MCP-1, IL-6, SREBF1 and ACC1 in liver tissue of db/db mice were all significantly increased. Western blot results showed that the expression levels of SREBF1, ACC1 increased, ferroptosis relative proteins were significantly decreased. The 8 weeks of exercise significantly reduced the rise in body weight, blood glucose, liver index and blood lipid levels in db/db mice. Exercise intervention also alleviated hepatic steatosis and reduced the expression levels of Fe2+, MDA, MCP-1, IL-6, ACC1 and SREBF1, upregulated the expression levels of GSH, NRF2, HO-1, SLC7A11 and GPX4 in liver tissue of db/db mice. The intervention of exercise combined with SB203580 significantly down-regulated the mRNA expression levels of ACC1, MCP-1, IL-6, reduced the levels of Fe2+ and MDA, and up-regulated the level of GSH in db/db mice. Compared with the db+Exe group, the expression of Fe2+, MDA, MCP-1, and SREBF1 in the liver of the db+Exe+SB203580 group mice significantly increased, while the expression level of GSH and expression levels of ferroptosis relative proteins also significantly decreased. In addition, compared with db+SB203580 group, the iron accumulation and lipid peroxidation in the liver of db+Exe+SB203580 group were significantly improved.Conclusion The 8-week treadmill exercise can effectively alleviate liver injury and steatosis, and its mechanism may be related to the inhibition of hepatocyte ferroptosis through p38 MAPK signal.]]> 2024/11/22 12:23:16 ZHANG Bao-Wen,LI Ying,GAO Yuan,SHENG Ke-Yan,WANG Zhi and KOU Xian-Juan 83true Objective Asthma is a common chronic inflammatory airway disease, and severe asthma poses a significant challenge in its diagnosis and management. Immune cells are involved in and altered by asthma pathogenesis, and several lipid metabolites can serve as diagnostic markers for the disease. In this study, we investigated the characterization of severe asthma at the metabolic and cellular level.Methods Differential metabolites in blood samples from severe asthma (41 cases) and controls (18 cases) were screened using multifactorial statistical analysis and independent samples t-tests; key pathways were identified by KEGG enrichment analysis, and biomarkers were characterized based on ROC curves; immune cell types and proportions in the blood were identified based on the results of cell-type annotations (5 severe and 3 control cases); and single-sample Gene Enrichment Analysis (ssGSEA) to investigate the characterization of differential metabolic pathways in single cells.Results Compared with controls, the abundance of 28 metabolites was increased and the abundance of 13 metabolites was decreased in the blood of patients with severe asthma (P<0.05); the differential metabolites were enriched in 4 pathways: sphingolipid metabolism, glycerophospholipid metabolism, nicotinate and nicotinamide metabolism, and histidine metabolism. Among them, 13 differential metabolites could be used as biomarkers for the diagnosis of severe asthma, including L-glutamic acid (AUC=0.809), nicotinamide (AUC=0.886), phytosphingosine (AUC=0.882), and sphinganine (AUC=0.893). In single-cell transcriptome analysis, 5 key cell types were identified: CD4+ T cells, CD8+ T cells, NK cells, B cells, and monocytes. The number of NK cells was increased in patients with severe asthma, and severe asthma exhibited more frequent cellular communication, particularly dense communication between CD8+ T cells and other cell types. In healthy samples, monocytes were the primary cells engaging in dense communication. Single-sample gene enrichment analysis (ssGSEA) showed that 4 pathways enriched for differential metabolites had lower scores (P<0.01) in CD4+ T and CD8+ T cells in severe patients, and it was hypothesized that the expression of genes associated with these pathways was suppressed in these two types of cells. The suppressed genes included DGKA and NT5C3A, which are associated with immune processes. We observed that these genes play key roles in the regulation of T cell signaling, activation, differentiation, and immune responses.Conclusion L-glutamic acid, nicotinamide, phytosphingosine, and sphinganine can be used as biomarkers for the diagnosis of severe asthma; genes of the severe asthma-associated pathway were suppressed in CD4+ T cells and CD8+ T cells.]]> 2024/11/22 12:23:25 JIANG Chen-Rong,CHEN Zhi-Hong and LIU Hong-De 82true Objective Coronavirus is a class of long-standing pathogens, which are enveloped single-stranded positive-sense RNA viruses. The genome all encodes 4 structural proteins: spike protein (S), nucleocapsid protein (N), membrane protein (M), and envelope protein (E). The nucleocapsid protein (NP) serves as a key structural component of coronaviruses, playing a vital function in the viral life cycle. NP acts as an RNA-binding protein, with a critical role in identifying specific sequences within the viral genome RNA, facilitating the formation of ribonucleoprotein (RNP) complexes with viral RNA to stabilize the viral genome and contribute to viral particles assembly. The NP consists of two primary structural domains, the N-terminal domain (NTD) and the C-terminal domain (CTD). The NTD is primarily responsible for RNA binding, whereas the CTD is involved in polymerization. The N protein demonstrated to trigger the host immune response and to modulate the cell cycle of infected cells by interacting with host proteins. The NP, one of the most abundant protein in coronaviruses, is essential in understanding the pathogenic mechanism of coronaviruses through its interaction with host factors, which response for determining the virus pathogenicity. HCoV-229E is a widely distributed coronavirus that typically causes mild upper respiratory tract diseases, accounting for a significant portion of common cold cases. However, its pathogenicity is notably lower compared to other coronaviruses like MERS-CoV, SARS-CoV, and SARS-CoV-2. The exact molecular mechanism behind remains unexplained, and how HCoV-229E N protein influences virus replication, host antiviral immunity, and pathogenesis need to be further explored.Methods Proximity labeling-mass spectrometry technique and bioinformatics analysis were used to screen for potential host factors interacting with the NP of human coronavirus 229E (HCoV-229E). In this study, a recombinant adenovirus Ad-V5-NPHCoV-229E-TurboID was constructed to express the fusion protein of HCoV-229E NP and biotin ligase (TurboID). A549 cells were infected with the Ad-V5-NPHCoV-229E-TurboID. After 30 min biotin treatment, NP interacting proteins were labeled with biotin by biotin ligase, and subsequently isolated with streptavidin cross-linked magnetic beads. The potential interacting proteins were identified using label-free proteomic mass spectrometry and further validated through immunoprecipitation and immunofluorescence assays.Results We identified a total of 584 potential interacting proteins. Gene ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis highlighted the enrichment of glycogen synthase kinase (GSK)3A and GSK3B in the glycolysis/gluconeogenesis pathway, indicating HCoV-229E NP connection to diabetes through aberrant activity. Moreover, SARS-CoV-2 infection can exacerbate hyperglycemia and metabolic dysregulation in diabetic individuals by activating the ACE2 receptor. Moreover, SARS-CoV-2 was observed to cause potentially harm to pancreatic β-cells and leading to insulin deficiency, which not only worsens the condition of diabetic patients but also raises the possibility of new-onset diabetes in non-diabetic individuals. We demonstrated that GSK3A and GSK3B interacted with NP of HCoV-229E, suggesting that the NP may engage in various coronavirus pathogenic processes by interacting with GSK3.Conclusion These findings suggest that proximity labeling-mass spectrometry technique is a valuable tool for identifying virus-host interaction factors, and lay the foundation for future investigations into the mechanisms underlying coronavirus replication, proliferation, and pathogenesis.]]> 2024/11/22 0:00:00 JU Rui-Xia,WANG Hao-Yong,LIU Hai-Nan,Liu Xuan and CAO Cheng 81true Objective As the central hub of the classical visual pathway, the primary visual cortex not only encodes and processes visual information but also establishes dense neural circuit connections with higher-order cognitive brain regions. Numerous studies have shown that 40 Hz flicker stimulation can induce γ oscillations in the brain and significantly improve learning and cognitive impairments in patients with neurodegenerative diseases. Moreover, flickering light phenomena naturally occur in daily environments. Given that the primary visual cortex serves as the brain’s first cortical hub for receiving visual input, it is essential to comprehensively understand how non-invasive light flicker stimulation modulates its information processing mechanisms. This study systematically investigates the effects of non-invasive light flicker stimulation at different frequencies on the functional properties of neurons in the primary visual cortex of adult mice, aiming to uncover how such stimulation modulates this region and, consequently, affects overall brain function.Methods Three groups of adult mice (approximately 12 weeks old) were exposed to light flicker stimulation at frequencies of 20 Hz, 40 Hz, and 60 Hz, respectively, for a duration of two months. A control group was exposed to the same light intensity without flickering. Following the stimulation period, in vivo multi-channel electrophysiological recordings were conducted. During these recordings, anesthetized mice were presented with various types of moving sinusoidal light gratings to assess the effects of different flicker frequencies on the functional properties of neurons in the primary visual cortex.Results The experimental results demonstrate that two months of light flicker stimulation at 20 Hz, 40 Hz, and 60 Hz enhances the orientation tuning capabilities of neurons in the primary visual cortex. Specifically, 40 Hz and 60 Hz stimulation improved contrast sensitivity, whereas 20 Hz had no significant effect. Further analysis revealed that all three frequencies reduced neuronal response variability (as measured by the Fano factor), increased the signal-to-noise ratio, and decreased noise correlation (rsc) between neurons.Conclusion Non-invasive light flicker stimulation enhances orientation tuning (e.g., orientation bias index) and contrast sensitivity (e.g., contrast threshold and C50) in neurons of the primary visual cortex. This enhancement is likely due to improved information processing efficiency, characterized by reduced neuronal variability and increased signal-to-noise ratio. These findings suggest that the primary visual cortex can achieve precise and efficient information encoding in complex lighting environments by selectively adapting to different flicker frequencies and optimizing receptive field properties. This study provides new experimental evidence on how various types of light flicker influence visual perception and offers insights into the mechanisms through which specific frequencies enhance brain function.]]> 2025/6/12 0:00:00 LI Xue-Qi,ZHOU Yi-Feng and XU Guang-Wei 80true Objective Traditional Chinese medicine (TCM) constitutes a valuable cultural heritage and an important source of antitumor compounds. Poria (Poria cocos (Schw.) Wolf), the dried sclerotium of a polyporaceae fungus, was first documented in Shennong’s Classic of Materia Medica and has been used therapeutically and dietarily in China for millennia. Traditionally recognized for its diuretic, spleen-tonifying, and sedative properties, modern pharmacological studies confirm that Poria exhibits antioxidant, anti-inflammatory, antibacterial, and antitumor activities. Pachymic acid (PA; a triterpenoid with the chemical structure 3β-acetyloxy-16α-hydroxy-lanosta-8,24(31)-dien-21-oic acid), isolated from Poria, is a principal bioactive constituent. Emerging evidence indicates PA exerts antitumor effects through multiple mechanisms, though these remain incompletely characterized. Neuroblastoma (NB), a highly malignant pediatric extracranial solid tumor accounting for 15% of childhood cancer deaths, urgently requires safer therapeutics due to the limitations of current treatments. Although PA shows multi-mechanistic antitumor potential, its efficacy against NB remains uncharacterized. This study systematically investigated the potential molecular targets and mechanisms underlying the anti-NB effects of PA by integrating network pharmacology-based target prediction with experimental validation of multi-target interactions through molecular docking, dynamic simulations, and in vitro assays, aimed to establish a novel perspective on PA’s antitumor activity and explore its potential clinical implications for NB treatment by integrating computational predictions with biological assays.Methods This study employed network pharmacology to identify potential targets of PA in NB, followed by validation using molecular docking, molecular dynamics (MD) simulations, MM/PBSA free energy analysis, RT-qPCR and Western blot experiments. Network pharmacology analysis included target screening via TCMSP, GeneCards, DisGeNET, SwissTargetPrediction, SuperPred, and PharmMapper. Subsequently, potential targets were predicted by intersecting the results from these databases via Venn analysis. Following target prediction, topological analysis was performed to identify key targets using Cytoscape software. Molecular docking was conducted using AutoDock Vina, with the binding pocket defined based on crystal structures. MD simulations were performed for 100 ns using GROMACS, and RMSD, RMSF, SASA, and hydrogen bonding dynamics were analyzed. MM/PBSA calculations were carried out to estimate the binding free energy of each protein-ligand complex. In vitro validation included RT-qPCR and Western blot, with GAPDH used as an internal control.Results The CCK-8 assay demonstrated a concentration-dependent inhibitory effect of PA on NB cell viability. GO analysis suggested that the anti-NB activity of PA might involve cellular response to chemical stress, vesicle lumen, and protein tyrosine kinase activity. KEGG pathway enrichment analysis suggested that the anti-NB activity of PA might involve the PI3K/AKT, MAPK, and Ras signaling pathways. Molecular docking and MD simulations revealed stable binding interactions between PA and the core target proteins AKT1, EGFR, SRC, and HSP90AA1. RT-qPCR and Western blot analyses further confirmed that PA treatment significantly decreased the mRNA and protein expression of AKT1, EGFR, and SRC while increasing the HSP90AA1 mRNA and protein levels.Conclusion It was suggested that PA may exert its anti-NB effects by inhibiting AKT1, EGFR, and SRC expression, potentially modulating the PI3K/AKT signaling pathway. These findings provide crucial evidence supporting PA’s development as a therapeutic candidate for NB.]]> 2025/6/16 0:00:00 LIU Hang,ZHU Yu-Xin,GUO Si-Lin,PAN Xin-Yun,XIE Yuan-Jie,LIAO Si-Cong,DAI Xin-Wen,SHEN Ping and XIAO Yu-Bo 79true Objective Tobacco-related diseases remain one of the leading preventable public health challenges worldwide and are among the primary causes of premature death. In recent years, accumulating evidence has supported the classification of nicotine addiction as a chronic brain disease, profoundly affecting both brain structure and function. Despite the urgency, effective diagnostic methods for smoking addiction remain lacking, posing significant challenges for early intervention and treatment. To address this issue and gain deeper insights into the neural mechanisms underlying nicotine dependence, this study proposes a novel graph neural network framework, termed TI-GNN. This model leverages functional magnetic resonance imaging (fMRI) data to identify complex and subtle abnormalities in brain connectivity patterns associated with smoking addiction.Methods The study utilizes fMRI data to construct functional connectivity matrices that represent interaction patterns among brain regions. These matrices are interpreted as graphs, where brain regions are nodes and the strength of functional connectivity between them serves as edges. The proposed TI-GNN model integrates a Transformer module to effectively capture global interactions across the entire brain network, enabling a comprehensive understanding of high-level connectivity patterns. Additionally, a spatial attention mechanism is employed to selectively focus on informative inter-regional connections while filtering out irrelevant or noisy features. This design enhances the model’s ability to learn meaningful neural representations crucial for classification tasks. A key innovation of TI-GNN lies in its built-in causal interpretation module, which aims to infer directional and potentially causal relationships among brain regions. This not only improves predictive performance but also enhances model interpretability—an essential attribute for clinical applications. The identification of causal links provides valuable insights into the neuropathological basis of addiction and contributes to the development of biologically plausible and trustworthy diagnostic tools.Results Experimental results demonstrate that the TI-GNN model achieves superior classification performance on the smoking addiction dataset, outperforming several state-of-the-art baseline models. Specifically, TI-GNN attains an accuracy of 0.91, an F1-score of 0.91, and a Matthews correlation coefficient (MCC) of 0.83, indicating strong robustness and reliability. Beyond performance metrics, TI-GNN identifies critical abnormal connectivity patterns in several brain regions implicated in addiction. Notably, it highlights dysregulations in the amygdala and the anterior cingulate cortex, consistent with prior clinical and neuroimaging findings. These regions are well known for their roles in emotional regulation, reward processing, and impulse control—functions that are frequently disrupted in nicotine dependence.Conclusion The TI-GNN framework offers a powerful and interpretable tool for the objective diagnosis of smoking addiction. By integrating advanced graph learning techniques with causal inference capabilities, the model not only achieves high diagnostic accuracy but also elucidates the neurobiological underpinnings of addiction. The identification of specific abnormal brain networks and their causal interactions deepens our understanding of addiction pathophysiology and lays the groundwork for developing targeted intervention strategies and personalized treatment approaches in the future.]]> 2025/7/11 16:16:00 WANG Xu-Wen,YU Da-Hua,XUE Ting,LI Xiao-Jiao,MAI Zhen-Zhen,DONG Fang,MA Yu-Xin,WANG Juan and YUAN Kai 78true Objective The proteome of biological evidence contains rich genetic information, namely single amino acid polymorphisms (SAPs) in protein sequences. However, due to the lack of efficient and convenient analysis tools, the application of SAP in public security still faces many challenges. This paper aims to meet the application requirements of SAP analysis for forensic biological evidence’s proteome data.Methods The software is divided into three modules. First, based on a built-in database of common non-synonymous single nucleotide polymorphisms (nsSNPs) and SAPs in East Asian populations, the software integrates and annotates newly identified exonic nsSNPs as SAPs, thereby constructing a customized SAP protein sequence database. It then utilizes a pre-installed search engine—either pFind or MaxQuant—to perform analysis and output SAP typing results, identifying both reference and variant types, along with their corresponding imputed nsSNPs. Finally, SAPTyper compares the proteome-based typing results with the individual’s exome-derived nsSNP profile and outputs the comparison report.Results SAPTyper accepts proteomic DDA mass spectrometry raw data (DDA acquisition mode) and exome sequencing results of nsSNPs as input and outputs the report of SAPs result. The pFind and Maxquant search engines were used to test the proteome data of 2 hair shafts of 2 individuals, and both obtained SAP results. It was found that the results of the Maxquant search engine were slightly less than those of pFind. This result shows that SAPTyper can achieve SAP fingding function. Moreover, the pFind search engine was used to test the proteome data of 3 hair shafts from 1 European person and 1 African person in the literature. Among the sites fully matched by the literature method, sites detected by SAPTyper are also included; for semi-matching sites, that is, nsSNPs are heterozygous, both literature method and SAPTyper method had the risk of missing detection for one type of the allele. Comparing the analysis results of SAPTyper with the SAP test results reported in the literature, it was found that some imputed nsSNP sites identified by the literature method but not detected by SAPTyper had a MAF of less than 0.1% in East Asian populations, and therefore they were not included in the common nsSNP database of East Asian populations constructed by this software. Since the database construction of this software is based on the genetic variation information of East Asian populations, it is currently unable to effectively identify representative unique common variation sites in European or African populations, but it can still identify SAP sites shared by these populations and East Asian populations.Conclusion An automated SAP analysis algorithm was developed for East Asian populations, and the software named SAPTyper was developed. This software provides a convenient and efficient analysis tool for the research and application of forensic proteomic SAP and has important application prospects in individual identification and phenotypic inference based on SAP.]]> 2025/7/2 9:26:55 HU Feng,WANG Meng-Jiao,WU Jia-Lei,DING Dong-Sheng,YANG Zhi-Yuan,JI An-Quan,FENG Lei and YE Jian 77true Objective Repetitive transcranial magnetic stimulation (rTMS), a non-invasive brain stimulation technique, offers a non-pharmacological therapeutic option for the management of Alzheimer’s disease (AD). Studies have demonstrated that ferroptosis plays a pivotal role in the pathological onset and progression of AD, and the inhibition of neuronal ferroptosis can significantly ameliorate cognitive impairments associated with AD. The imbalance of calcium ion (Ca2+) homeostasis is intimately associated with the pathology of AD and serves as a catalyst for the induction of ferroptosis through various pathways. This study is designed to investigate whether rTMS can ameliorate AD by inhibiting neuronal ferroptosis or maintaining calcium homeostasis, ultimately establishing a theoretical and experimental framework for the utilization of rTMS in AD treatment.Methods APP/PS1 AD mice were subjected to both 0.5 Hz low-frequency and 20 Hz high-frequency rTMS treatments, and the efficacy of these treatments was evaluated using novel object recognition and Morris water maze tests. ELISA was employed to quantify the levels of glutathione (GSH), malondialdehyde (MDA), superoxide dismutase (SOD), Fe2+ within the hippocampi of mice from each group. HT-22 cells were induced to undergo ferroptosis via Erastin treatment, and subsequent to high- and low-frequency magnetic stimulation, cell viability was assessed using CCK-8 assay, while intracellular calcium ion concentration fluctuations were monitored using Fluo-4 AM.Results The findings revealed that, when compared to normal mice, AD mice displayed a notable decline in cognitive function, accompanied by a substantial increase in ferroptosis levels and intracellular calcium ion concentrations. Both high-frequency and low-frequency applications of rTMS were found to significantly ameliorate cognitive impairments in AD mice, while also effectively mitigating the abnormal augmentation of neuronal ferroptosis and intracellular calcium ion levels.Conclusion The present study underscores that both high-frequency and low-frequency rTMS exhibit efficacy in alleviating cognitive dysfunction in AD mice, potentially through the modulation of ferroptosis and intracellular calcium ion homeostasis.]]> 2025/4/30 11:25:34 ZHANG Meng, ZHANG Ze, FU Rui, REN Zi-Hao, DING Chong 76true Objective Based on functional near-infrared spectroscopy (fNIRS), we investigated the brain activity characteristics of gross motor tasks in children with autism spectrum disorder (ASD) and motor dysfunctions (MDs) to provide a theoretical basis for further understanding the mechanism of MDs in children with ASD and designing targeted intervention programs from a central perspective.Methods According to the inclusion and exclusion criteria, 48 children with ASD accompanied by MDs were recruited into the ASD group and 40 children with typically developing (TD) into the TD group. The fNIRS device was used to collect the information of blood oxygen changes in the cortical motor-related brain regions during single-handed bag throwing and tiptoe walking, and the differences in brain activation and functional connectivity between the two groups of children were analyzed from the perspective of brain activation and functional connectivity.Results Compared to the TD group, in the object manipulative motor task (one-handed bag throwing), the ASD group showed significantly reduced activation in both left sensorimotor cortex (SMC) and right secondary visual cortex (V2) (P<0.05), whereas the right pre-motor and supplementary motor cortex (PMC&SMA) had significantly higher activation (P<0.01) and showed bilateral brain region activity; in terms of brain functional integration, there was a significant decrease in the strength of brain functional connectivity (P<0.05) and was mainly associated with dorsolateral prefrontal cortex (DLPFC) and V2. In the body stability motor task (tiptoe walking), the ASD group had significantly higher activation in motor-related brain regions such as the DLPFC, SMC, and PMC&SMA (P<0.05) and showed bilateral brain region activity; in terms of brain functional integration, the ASD group had lower strength of brain functional connectivity (P<0.05) and was mainly associated with PMC&SMA and V2.Conclusion Children with ASD exhibit abnormal brain functional activity characteristics specific to different gross motor tasks in object manipulative and body stability, reflecting insufficient or excessive compensatory activation of local brain regions and impaired cross-regions integration, which may be a potential reason for the poorer gross motor performance of children with ASD, and meanwhile provides data support for further unraveling the mechanisms underlying the occurrence of MDs in the context of ASD and designing targeted intervention programs from a central perspective.]]> 2025/5/22 15:10:56 ZONG Wen-Hao, LIANG Qi, YANG Shi-Yu, WANG Feng-Jiao, WEI Meng-Zhao, LEI Hong, DONG Gui-Jun, LI Ke-Feng 75true Objective The nucleolar protein PES1 (Pescadillo homolog 1) plays critical roles in ribosome biogenesis and cell cycle regulation, yet its involvement in cellular senescence remains poorly understood. This study aimed to comprehensively investigate the functional consequences of PES1 suppression in cellular senescence and elucidate the molecular mechanisms underlying its regulatory role.Methods Initially, we assessed PES1 expression patterns in two distinct senescence models: replicative senescent mouse embryonic fibroblasts (MEFs) and doxorubicin-induced senescent human hepatocellular carcinoma HepG2 cells. Subsequently, PES1 expression was specifically downregulated using siRNA-mediated knockdown in these cell lines as well as additional relevant cell types. Cellular proliferation and senescence were assessed by EdU incorporation and SA-β-gal staining assays, respectively. The expression of senescence-associated proteins (p53, p21, and Rb) and SASP factors (IL-6, IL-1β, and IL-8) were analyzed by Western blot or qPCR. Furthermore, Northern blot and immunofluorescence were employed to evaluate pre-rRNA processing and nucleolar morphology.Results PES1 expression was significantly downregulated in senescent MEFs and HepG2 cells. PES1 knockdown resulted in decreased EdU-positive cells and increased SA-β-gal-positive cells, indicating proliferation inhibition and senescence induction. Mechanistically, PES1 suppression activated the p53-p21 pathway without affecting Rb expression, while upregulating IL-6, IL-1β, and IL-8 production. Notably, PES1 depletion impaired pre-rRNA maturation and induced nucleolar stress, as evidenced by aberrant nucleolar morphology.Conclusion Our findings demonstrate that PES1 deficiency triggers nucleolar stress and promotes p53-dependent (but Rb-independent) cellular senescence, highlighting its crucial role in maintaining nucleolar homeostasis and regulating senescence-associated pathways.]]> 2025/4/15 13:44:27 ZHANG Chang-Jian,LI Yu-Fang,WU Feng-Yun,JIN Rui,NIU Chang,YE Qi-Nong and CHENG Long 74true Objective This paper proposes a novel real-time bedside pulmonary ventilation monitoring method for the diagnosis of chronic obstructive pulmonary disease (COPD), based on electrical impedance tomography (EIT). Four indicators—center of ventilation (CoV), global inhomogeneity index (GI), regional ventilation delay inhomogeneity (RVDI), and the ratio of forced expiratory volume in one second to forced vital capacity (FEV1/FVC)—are calculated to enable the spatiotemporal assessment of COPD.Methods A simulation of the respiratory cycles of COPD patients was first conducted, revealing significant differences in certain indicators compared to healthy individuals. The effectiveness of these indicators was then validated through experiments. A total of 93 subjects underwent multiple pulmonary function tests (PFTs) alongside simultaneous EIT measurements. Ventilation heterogeneity under different breathing patterns—including forced exhalation, forced inhalation, and quiet tidal breathing—was compared. EIT images and related indicators were analyzed to distinguish healthy individuals across different age groups from COPD patients.Results Simulation results demonstrated significant differences in CoV, GI, FEV1/FVC, and RVDI between COPD patients and healthy individuals. Experimental findings indicated that, in terms of spatial heterogeneity, the GI values of COPD patients were significantly higher than those of the other two groups, while no significant differences were observed among healthy individuals. Regarding temporal heterogeneity, COPD patients exhibited significantly higher RVDI values than the other groups during both quiet breathing and forced inhalation. Moreover, during forced exhalation, the distribution of FEV1/FVC values further highlighted the temporal delay heterogeneity of regional lung function in COPD patients, distinguishing them from healthy individuals of various ages.Conclusion EIT technology effectively reveals the spatiotemporal heterogeneity of regional lung function, which holds great promise for the diagnosis and management of COPD.]]> 2025/4/15 13:48:12 LI Fang,CHEN Bai,WU Yang,LIU Kai,ZHOU Tong and YAO Jia-Feng 73true Objective The aim of this study was to investigate the prophylactic effects of caloric restriction (CR) on lipopolysaccharide (LPS)-induced septic cardiomyopathy (SCM) and to elucidate the mechanisms underlying the cardioprotective actions of CR. This research aims to provide innovative strategies and theoretical support for the prevention of SCM.Methods A total of forty-eight 8-week-old male C57BL/6 mice, weighing between 20-25 g, were randomly assigned to 4 distinct groups, each consisting of 12 mice. The groups were designated as follows: CON (control), LPS, CR, and CR+LPS. Prior to the initiation of the CR protocol, the CR and CR+LPS groups underwent a 2-week acclimatization period during which individual food consumption was measured. The initial week of CR intervention was set at 80% of the baseline intake, followed by a reduction to 60% for the subsequent 5 weeks. After 6-week CR intervention, all 4 groups received an intraperitoneal injection of either normal saline or LPS (10 mg/kg). Twelve hours post-injection, heart function was assessed, and subsequently, heart and blood samples were collected. Serum inflammatory markers were quantified using enzyme-linked immunosorbent assay (ELISA). The serum myocardial enzyme spectrum was analyzed using an automated biochemical instrument. Myocardial tissue sections underwent hematoxylin and eosin (HE) staining and immunofluorescence (IF) staining. Western blot analysis was used to detect the expression of protein in myocardial tissue, including inflammatory markers (TNF-α, IL-9, IL-18), oxidative stress markers (iNOS, SOD2), pro-apoptotic markers (Bax/Bcl-2 ratio, CASP3), and SIRT3/SIRT6.Results Twelve hours after LPS injection, there was a significant decrease in ejection fraction (EF) and fractional shortening (FS) ratios, along with a notable increase in left ventricular end-systolic diameter (LVESD). Morphological and serum indicators (AST, LDH, CK, and CK-MB) indicated that LPS injection could induce myocardial structural disorders and myocardial injury. Furthermore, 6-week CR effectively prevented the myocardial injury. LPS injection also significantly increased the circulating inflammatory levels (IL-1β, TNF-α) in mice. IF and Western blot analyses revealed that LPS injection significantly up-regulating the expression of inflammatory-related proteins (TNF-α, IL-9, IL-18), oxidative stress-related proteins (iNOS, SOD2) and apoptotic proteins (Bax/Bcl-2 ratio, CASP3) in myocardial tissue. 6-week CR intervention significantly reduced circulating inflammatory levels and downregulated the expression of inflammatory, oxidative stress-related proteins and pro-apoptotic level in myocardial tissue. Additionally, LPS injection significantly downregulated the expression of SIRT3 and SIRT6 proteins in myocardial tissue, and CR intervention could restore the expression of SIRT3 proteins.Conclusion A 6-week CR could prevent LPS-induced septic cardiomyopathy, including cardiac function decline, myocardial structural damage, inflammation, oxidative stress, and apoptosis. The mechanism may be associated with the regulation of SIRT3 expression in myocardial tissue.]]> 2024/10/29 22:52:23 ZHANG Ming-Chen,ZHANG Hui,LI Ting-Ting,CHEN Ming-Hua,WANG Xiao-Wen and SUN Zhong-Guang 72true Objective Functional near-infrared spectroscopy (fNIRS), a novel non-invasive technique for monitoring cerebral activity, can be integrated with upper limb rehabilitation robots to facilitate the real-time assessment of neurological rehabilitation outcomes. The rehabilitation robot is designed with 3 training modes: passive, active, and resistance. Among these, the resistance mode has been demonstrated to yield superior rehabilitative outcomes for patients with a certain level of muscle strength. The control modes in the resistance mode can be categorized into dynamic and static control. However, the effects of different control modes in the resistance mode on the motor function of patients with upper limb hemiplegia in stroke remain unclear. Furthermore, the effects of force, an important parameter of different control modes, on the activation of brain regions have rarely been reported. This study investigates the effects of dynamic and static resistance modes under varying resistance levels on cerebral functional alterations during motor rehabilitation in post-stroke patients.Methods A cohort of 20 stroke patients with upper limb dysfunction was enrolled in the study, completing preparatory adaptive training followed by 3 intensity-level tasks across 2 motor paradigms. The bilateral prefrontal cortices (PFC), bilateral primary motor cortices (M1), bilateral primary somatosensory cortices (S1), and bilateral premotor and supplementary motor cortices (PM) were examined in both the resting and motor training states. The lateralization index (LI), phase locking value (PLV), network metrics were employed to examine cortical activation patterns and topological properties of brain connectivity.Results The data indicated that both dynamic and static modes resulted in significantly greater activation of the contralateral M1 area and the ipsilateral PM area when compared to the resting state. The static patterns demonstrated a more pronounced activation in the contralateral M1 in comparison to the dynamic patterns. The results of brain network analysis revealed significant differences between the dynamic and resting states in the contralateral PFC area and contralateral M1 area (F=4.709, P=0.038), as well as in the contralateral PM area and ipsilateral M1 area (F=4.218, P=0.049). Moreover, the findings indicated a positive correlation between the activation of the M1 region and the increase in force in the dynamic mode, which was reversed in the static mode.Conclusion Both dynamic and static resistance training modes have been demonstrated to activate the corresponding brain functional regions. Dynamic resistance modes elicit greater oxygen changes and connectivity to the region of interest (ROI) than static resistance modes. Furthermore, the effects of increasing force differ between the two modes. In patients who have suffered a stroke, dynamic modes may have a more pronounced effect on the activation of exercise-related functional brain regions.]]> 2025/4/1 15:20:05 FU Ling-Di,DOU Jia-Xuan,YING Ting-Ting,YIN Li-Yong,TANG Min and LIANG Zhen-Hu 71true Objective This study sought to investigate the impact of exosomes derived from LoVo cells (LoVo-Exos) in colorectal cancer (CRC) on tumor angiogenesis, as well as to elucidate the potential molecular mechanisms underlying their pro-angiogenic effects.Methods LoVo-Exos were isolated via ultracentrifugation, and their internalization into recipient human umbilical vein endothelial cells (HUVECs) was visualized using confocal microscopy. The influence of LoVo-Exos on angiogenesis was assessed through an in vitro tube formation assay. Additionally, the pro-angiogenic effects of LoVo-Exos were evaluated in vivo using a matrix gluing assay in mice. To investigate the molecular mechanisms through which LoVo-Exos facilitate angiogenesis, Western blot analysis was employed to examine the transfer of pEGFR by LoVo-Exos into recipient cells. Both Western blot and ELISA were utilized to assess the expression levels of key signaling proteins within the EGFR-ERK pathway, as well as the expression of downstream angiogenic core molecules. Furthermore, the impact of EGFR knockdown and ERK inhibitor treatment on angiogenesis was evaluated, with subsequent analysis of the expression of downstream angiogenic core molecules following these interventions.Results Confocal microscopy demonstrated the internalization of LoVo-Exos into HUVECs. In vitro angiogenesis assays further indicated that LoVo-Exos significantly enhanced the formation of tubular structures in HUVECs. Additionally, macroscopic examination of subcutaneous matrix plug formation in mice revealed a substantial increase in vascular-like structures within the matrix plugs following the administration of LoVo-Exos, compared to the PBS control group. Hematoxylin and eosin (HE) staining revealed the presence of erythrocyte-filled microvessels within the matrix plugs combined with LoVo-Exos. Furthermore, immunohistochemical analysis demonstrated the expression of the endothelial cell marker CD31 in these matrix plugs. The presence of CD31-positive cells in the LoVo-Exos-treated matrix plugs was associated with a significant enhancement in the formation of luminal structures. These findings suggest that LoVo-Exos facilitate the in vivo development of vascular-like structures. Subsequent investigations demonstrated that LoVo-Exos facilitated the delivery of pEGFR to HUVEC, thereby enhancing angiogenesis. Conversely, LoVo-Exos with EGFR knockdown exhibited a diminished capacity to promote angiogenesis, an effect that was further attenuated by the ERK phosphorylation inhibitor U0126. Western blot analysis assessing the activation of the EGFR-ERK signaling pathway in HUVEC indicated that LoVo-Exos augmented angiogenesis through the activation of this pathway. Furthermore, analysis of the impact of LoVo-Exos on the expression of downstream angiogenic core molecules revealed an increase in interleukin-8 (IL-8) secretion in HUVEC. The enhancement observed was diminished in LoVo-Exos following EGFR knockdown, and this reduction was counteracted by the ERK phosphorylation inhibitor U0126.Conclusion The underlying mechanism may involve the delivery of pEGFR in LoVo-Exos to HUVECs, leading to increased IL-8 secretion via the EGFR-ERK signaling pathway, thereby enhancing the angiogenic potential of HUVECs. This finding may offer new insights into the mechanisms underlying cancer metastasis.]]> 2025/1/9 10:10:10 CHENG Ya-Jie,ZHOU Xue-Tong,WANG Rui and FANG Jin 70true Objective Hepatocyte nuclear factor 4-alpha (HNF4A) is a critical transcription factor in the liver and pancreas. Dysfunctions of HNF4A lead to maturity onset diabetes of the young 1 (MODY1). Notably, MODY1 patients with HNF4A pathogenic mutations exhibit decreased responses to arginine and reduced plasma triglyceride levels, but the mechanisms remain unclear. This study aims to investigate the potential target genes transcriptionally regulated by HNF4A and explore its role in these metabolic pathways.Methods A stable 293T cell line expressing the HNF1A reporter was overexpressed with HNF4A. RNA sequencing (RNA-seq) was performed to analyze transcriptional differences. Transcription factor binding site prediction was then conducted to identify HNF4A binding motifs in the promoter regions of relevant target genes.Results RNA-seq results revealed a significant upregulation of transmembrane 4 L six family member 5 (TM4SF5) mRNA in HNF4A-overexpressing cells. Transcription factor binding predictions suggested the presence of five potential HNF4A binding motifs in the TM4SF5 promoter. Finally, we confirmed that the DR1 site in the -57 to -48 region of the TM4SF5 promoter is the key binding motif for HNF4A.Conclusion This study identified TM4SF5 as a target gene of HNF4A and determined the key binding motif involved in its regulation. Given the role of TM4SF5 as an arginine sensor in mTOR signaling activation and triglyceride secretion, which closely aligns with phenotypes observed in MODY1 patients, our findings provide novel insights into the possible mechanisms by which HNF4A regulates triglyceride secretion in the liver and arginine-stimulated insulin secretion in the pancreas.]]> 2025/2/12 0:00:00 GUO Yi-Ming,ZHANG Xiao-Fei,FENG Han and ZHENG Li 69true Objective As a second messenger in intracellular signal transduction, Ca2+ plays an important role in cell migration. Previous studies have demonstrated that extracellular Ca2+ influx can promote electric field-guided cell migration, known as electrotaxis. However, the effect of intracellular Ca2+ flow on electrotaxis is unclear. Therefore, in this study, we investigate the effect of Ca2+ flux on the electrotaxis of Dictyostelium discoideum.Methods The electrotaxis of Dictyostelium discoideum was investigated by applying a direct current (DC) electric field. Cell migration was recorded using a real-time imaging system. Calcium channel inhibitors, the extracellular Ca2+ chelator EGTA, Ca2+-free DB buffer, and caffeine were applied to investigate the impact of intra- and extracellular Ca2+ flow on electrotaxis. The involvement of G proteins and ERK2 in directed cell migration mediated by endoplasmic reticulum Ca2+ release was explored using mutants.Results Dictyostelium discoideum migrated toward the cathode in the electric field in a voltage-dependent manner. The intracellular Ca2+ concentration of the cells was significantly increased in the electric field. Inhibition of both extracellular Ca2+ influx and intracellular Ca2+ release suppressed cell electrotaxis migration. Inhibition of endoplasmic reticulum Ca2+ release induced by caffeine significantly impaired the electrotaxis of Dictyostelium discoideum. Deletion of Gα2, Gβ, Gγ, and Erk2 notably reduced the electrotaxis of the cells. Enhancing Ca2+ release mediated by caffeine restored the electrotaxis of the Gα2-, Gβ -, and Erk2- mutant cells partially or completely, but did not restore electrotaxis in the Gγ- mutant cells.Conclusion Ca2+ release from the endoplasmic reticulum regulates electrotaxis migration in Dictyostelium discoideum and is involved in the regulation of cell electrotaxis by G proteins and ERK2.]]> 2025/1/8 15:32:21 WANG Yi-Fan,YUAN Shu-Qin,GAO Run-Chi and ZHAO San-Jun 68true Objective Non-invasive magnetic stimulation technology has been widely used in the treatment of Alzheimer’s disease (AD), but there is a lack of convenient and timely methods for evaluating and providing feedback on the effectiveness of the stimulation, which can be used to guide the adjustment of the stimulation protocol. This study aims to explore the possibility of heart rate variability (HRV) in diagnosing AD and guiding AD magnetic stimulation intervention techniques.Methods In this study, we used a 40 Hz, 10 mT pulsed magnetic field to expose AD mouse models to whole-body exposure for 18 d, and detected the behavioral and electroencephalographic signals before and after exposure, as well as the instant electrocardiographic signals after exposure every day.Results Using one-way ANOVA and Pearson correlation coefficient analysis, we found that some HRV indicators could identify AD mouse models as accurately as behavioral and electroencephalogram(EEG) changes (P<0.05) and significantly distinguish the severity of the disease (P<0.05), including rMSSD, pNN6, LF/HF, SD1/SD2, and entropy arrangement. These HRV indicators showed good correlation and statistical significance with behavioral and EEG changes (r>0.3, P<0.05); HRV indicators were significantly modulated by the magnetic field exposure before and after the exposure, both of which were observed in the continuous changes of electrocardiogram (ECG) (P<0.05), and the trend of the stimulation effect was more accurately observed in the continuous changes of ECG.Conclusion HRV can accurately reflect the pathophysiological changes and disease degree, quickly evaluate the effect of magnetic stimulation, and has the potential to guide the pattern of magnetic exposure, providing a new idea for the study of personalized electromagnetic neuroregulation technology for brain diseases.]]> 2025/2/24 17:38:54 CHEN Shu-Ting,GENG Du-Yan,FAN Chun-Meng,ZHENG Wei-Ran and XU Gui-Zhi 67true Objective The central symptom of Parkinson’s disease (PD) is impaired motor function. Beta-band electrical activity in the motor network of the basal ganglia is closely related to motor function. In this study, we combined scalp electroencephalography (EEG), brain functional network, and clinical scales to investigate the effects of beta burst-period neural electrical activity on brain functional network characteristics, which may serve as a reference for clinical diagnosis and treatment.Methods Thirteen PD patients were included in the PD group, and 13 healthy subjects were included in the healthy control group. Resting-state EEG data were collected from both groups, and beta burst and non-burst periods were extracted. A phase synchronization network was constructed using weighted phase lag indices, and the topological feature parameters of phase synchronization network were compared between the two groups across different periods and four frequency bands. Additionally, the correlation between changes in network characteristics and clinical symptoms was analyzed.Results During the beta burst period, the topological characteristic parameters of phase synchronization network in all four frequency bands were significantly higher in PD patients compared to healthy controls. The average clustering coefficient of the phase synchronization network in the beta band during the beta burst period was negatively correlated with UPDRS-III scores. In the low gamma band during the non-burst period, the average clustering coefficient of phase synchronization network was positively correlated with UPDRS and UPDRS-III scores, while UPDRS-III scores were positively correlated with global efficiency and average degree.Conclusion The brain functional network features of PD patients were significantly enhanced during the beta burst period. Moreover, the beta-band brain functional network characteristics during the beta burst period were negatively correlated with clinical scale scores, whereas low gamma-band functional network features during the non-burst period were positively correlated with clinical scale scores. These findings indicate that motor function impairment in PD patients is associated with the beta burst period. This study provides valuable insights for the diagnosis of PD.]]> 2025/2/28 11:13:03 HAO Yu-Jie,YANG Shuo,LIU Shuo,LOU Xu and WANG Lei 66true Objective Bola-like short peptides exhibit novel self-assembly properties due to the formation of peptide dimers via hydrogen bonding interactions between their C-terminals. In this configuration, hydrophilic amino acids are distributed at both terminals, making these peptides behave similarly to Bola peptides. The electrostatic repulsive interactions arising from the hydrophilic amino acids at each terminal can be neutralized, thereby greatly promoting the lateral association of β-sheets. Consequently, assemblies with significantly larger widths are typically the dominant nanostructures for Bola-like peptides. To investigate the effect of hydrophilic amino acids on the self-assembly behavior of Bola-like peptides, the peptides Ac-RI3-CONH2 and Ac-HI3-CONH2 were designed and synthesized using the Bola-like peptide Ac-KI3-CONH2 as a template. Their self-assembly behavior was systematically examined.Methods Atomic force microscopy (AFM) and transmission electron microscopy (TEM) were employed to characterize the morphology and size of the assemblies. The secondary structures of the assemblies were analyzed using circular dichroism (CD) and Fourier transform infrared (FTIR) spectroscopy. Small-angle neutron scattering (SANS) was used to obtain detailed structural information at a short-length scale. Based on these experimental results, the effects of hydrophilic amino acids on the self-assembly behavior of Bola-like short peptides were systematically analyzed, and the underlying formation mechanism was explored.Results The aggregation process primarily involved three steps. First, peptide dimers were formed through hydrogen bonding interactions between their C-terminals. Within these dimers, the hydrophilic amino acids K, R, and H were positioned at both terminals, enabling the peptides to self-assemble in a manner similar to Bola peptides. Next, β-sheets were formed via hydrogen bonding interactions along the peptide backbone. Finally, self-assemblies were generated through the lateral association of β-sheets. The results demonstrated that both Ac-KI3-CONH2 and Ac-RI3-CONH2 could self-assemble into double-layer nanotubes with diameters of approximately 200 nm. These nanotubes were formed by the edge fusion of helical ribbons, which initially emerged from twisted ribbons. Notably, the primary assemblies of these peptides exhibited opposite chirality: nanofibers formed by Ac-KI3-CONH2 displayed left-handed chirality, whereas those formed by Ac-RI3-CONH2 exhibited right-handed chirality. This reversal in torsional direction was primarily attributed to the different abilities of K and R to form hydrogen bonds with water. In contrast, Ac-HI3-CONH2 formed narrower twisted ribbons with a significantly reduced width of approximately 30 nm, which was attributed to the strong steric hindrance caused by the imidazole rings. The multilayer height of these ribbons was mainly due to the unique structure of the imidazole rings, which can function as both hydrogen bond donors and acceptors, thereby promoting aggregate growth in the vertical direction.Conclusion The final morphology of the self-assemblies resulted from a delicate balance of various non-covalent interactions. By altering the types of hydrophilic amino acid residues in Bola-like short peptides, the relative strength of non-covalent interactions that drive assembly formation can be effectively regulated, allowing precise control over the morphology and chirality of the assemblies. This study provides a simple and effective approach for constructing diverse self-assemblies and lays a theoretical foundation for the development of functional biomaterials.]]> 2025/2/14 0:00:00 GAO Xin-Xin,HAN Yu,ZHOU Yi-Lin,CHEN Xi-Ya and ZHAO Yu-Rong 65true Objective The rapid advancement of bioanalytical technologies has heightened the demand for high-throughput, label-free, and real-time cellular analysis. Electrochemical impedance spectroscopy (EIS) operating in the GHz frequency range (GHz-EIS) has emerged as a promising tool for characterizing cell suspensions due to its ability to rapidly and non-invasively capture the dielectric properties of cells and their microenvironment. Although GHz-EIS enables rapid and label-free detection of cell suspensions, significant challenges remain in interpreting GHz impedance data for complex samples, limiting the broader application of this technique in cellular research. To address these challenges, this study presents a novel method that integrates GHz-EIS with deep learning algorithms, aiming to improve the precision of cell suspension concentration identification and quantification. This method provides a more efficient and accurate solution for the analysis of GHz impedance data.Methods The proposed method comprises two key components: dielectric property dataset construction and Backpropagation (BP) Neural Network modeling. Yeast cell suspensions at varying concentrations were prepared and separately introduced into a coaxial sensor for impedance measurement. The dielectric properties of these suspensions were extracted using a GHz-EIS dielectric property extraction method applied to the measured impedance data. A dielectric properties dataset incorporating concentration labels was subsequently established and divided into training and testing subsets. A BP neural network model employing specific activation functions (ReLU and Leaky ReLU) was then designed. The model was trained and tested using the constructed dataset, and optimal model parameters were obtained through this process. This BP neural network enables automated extraction and analytical processing of dielectric properties, facilitating precise recognition of cell suspension concentrations through data-driven training.Results Through comparative analysis with conventional centrifugal methods, the recognized concentration values of cell suspensions showed high consistency, with relative errors consistently below 5%. Notably, high-concentration samples exhibited even smaller deviations, further validating the precision and reliability of the proposed methodology. To benchmark the recognition performance against different algorithms, two typical approaches—Support Vector Machines (SVM) and K-Nearest Neighbor (KNN)—were selected for comparison. The proposed method demonstrated superior performance in quantifying cell concentrations. Specifically, the BP neural network achieved a mean absolute percentage error (MAPE) of 2.06% and an R2 value of 0.997 across the entire concentration range, demonstrating both high predictive accuracy and excellent model fit.Conclusion This study demonstrates that the proposed method enables accurate and rapid determination of unknown sample concentrations. By combining GHz-EIS with BP neural network algorithms, efficient identification of cell concentrations is achieved, laying the foundation for the development of a convenient online cell analysis platform and showing significant application prospects. Compared to typical recognition approaches, the proposed method exhibits superior capabilities in recognizing cell suspension concentrations. Furthermore, this methodology not only accelerates research in cell biology and precision medicine but also paves the way for future EIS biosensors capable of intelligent, adaptive analysis in dynamic biological research.]]> 2025/2/27 16:40:06 ZHANG An,TAO A-Long,RAN Qi-Hang,LIU Xia-Yi,WANG Zhi-Long,SUN Bo,YAO Jia-Feng and ZHAO Tong 64true Objective This study leverages structural data from antigen-antibody complexes of the influenza A virus neuraminidase (NA) protein to investigate the spatial recognition relationship between the antigenic epitopes and antibody paratopes.Methods Structural data on NA protein antigen-antibody complexes were comprehensively collected from the SAbDab database, and processed to obtain the amino acid sequences and spatial distribution information on antigenic epitopes and corresponding antibody paratopes. Statistical analysis was conducted on the antibody sequences, frequency of use of genes, amino acid preferences, and the lengths of complementarity determining regions (CDR). Epitope hotspots for antibody binding were analyzed, and the spatial structural similarity of antibody paratopes was calculated and subjected to clustering, which allowed for a comprehensively exploration of the spatial recognition relationship between antigenic epitopes and antibodies. The specificity of antibodies targeting different antigenic epitope clusters was further validated through bio-layer interferometry (BLI) experiments.Results The collected data revealed that the antigen-antibody complex structure data of influenza A virus NA protein in SAbDab database were mainly from H3N2, H7N9 and H1N1 subtypes. The hotspot regions of antigen epitopes were primarily located around the catalytic active site. The antibodies used for structural analysis were primarily derived from human and murine sources. Among murine antibodies, the most frequently used V-J gene combination was IGHV1-12*01/IGHJ2*01, while for human antibodies, the most common combination was IGHV1-69*01/IGHJ6*01. There were significant differences in the lengths and usage preferences of heavy chain CDR amino acids between antibodies that bind within the catalytic active site and those that bind to regions outside the catalytic active site. The results revealed that structurally similar antibodies could recognize the same epitopes, indicating a specific spatial recognition between antibody and antigen epitopes. Structural overlap in the binding regions was observed for antibodies with similar paratope structures, and the competitive binding of these antibodies to the epitope was confirmed through BLI experiments.Conclusion The antigen epitopes of NA protein mainly ditributed around the catalytic active site and its surrounding loops. Spatial complementarity and electrostatic interactions play crucial roles in the recognition and binding of antibodies to antigenic epitopes in the catalytic region. There existed a spatial recognition relationship between antigens and antibodies that was independent of the uniqueness of antibody sequences, which means that antibodies with different sequences could potentially form similar local spatial structures and recognize the same epitopes.]]> 2024/12/4 10:35:01 ZHU Zheng,CHEN Zheng-Shan,ZHANG Guan-Ying,FANG Ting,FAN Pu,BI Lei,CUI Yue,LI Ze-Ya,SU Chun-Yi,CHI Xiang-Yang and YU Chang-Ming 63true Objective In the realm of forensic science, dust is a valuable type of trace evidence with immense potential for intricate investigations. With the development of DNA sequencing technologies, there is a heightened interest among researchers in unraveling the complex tapestry of microbial communities found within dust samples. Furthermore, striking disparities in the microbial community composition have been noted among dust samples from diverse geographical regions, heralding new possibilities for geographical inference based on microbial DNA analysis. The pivotal role of microbial community data from dust in geographical inference is significant, underscoring its critical importance within the field of forensic science. This study aims to delve deeply into the nuances of fungal community composition across the urban landscapes of Beijing, Fuzhou, Kunming, and Urumqi in China. It evaluates the accuracy of biogeographic inference facilitated by the internal transcribed spacer 2 (ITS2) fungal sequencing while concurrently laying a robust foundation for the operational integration of environmental DNA into geographical inference mechanisms.Methods ITS2 region of the fungal genomes was amplified using universal primers known as 5.8S-Fun/ITS4-Fun, and the resulting DNA fragments were sequenced on the Illumina MiSeq FGx platform. Non-metric multidimensional scaling analysis (NMDS) was employed to visually represent the differences between samples, while analysis of similarities (ANOSIM) and permutational multivariate analysis of variance (PERMANOVA) were utilized to statistically evaluate the dissimilarities in community composition across samples. Furthermore, using Linear Discriminant Analysis Effect Size (LEfSe) analysis to identify and filter out species that exhibit significant differences between various cities. In addition, we leveraged SourceTracker to predict the geographic origins of the dust samples.Results Among the four cities of Beijing, Fuzhou, Kunming and Urumqi, Beijing has the highest species richness. The results of species annotation showed that there were significant differences in the species composition and relative abundance of fungal communities in the four cities. NMDS analysis revealed distinct clustering patterns of samples based on their biogeographic origins in multidimensional space. Samples from the same city exhibited clear clustering, while samples from different cities showed separation along the first axis. The results from ANOSIM and PERMANOVA confirmed the significant differences in fungal community composition between the four cities, with the most pronounced distinctions observed between Fuzhou and Urumqi. Notably, the biogeographic origins of all known dust samples were successfully predicted.Conclusion Significant differences are observed in the fungal species composition and relative abundance among the cities of Beijing, Fuzhou, Kunming, and Urumqi. Employing fungal ITS2 sequencing on dust samples from these urban areas enables accurate inference of biogeographical locations. The high feasibility of utilizing fungal community data in dust for biogeographical inferences holds particular promise in the field of forensic science.]]> 2024/11/24 23:03:07 ZHANG Wen-Jun,FENG Yao-Sen,PENG Jia-Jin,FENG Kai,Deng Ye,KANG Ke-Lai and WANG Le 62true Objective A multiplex amplification system was constructed based on the capillary electrophoresis platform for simultaneous detection of saliva, semen, and vaginal secretions using tissue-specific RNA markers. The aim of this study is to identify the tissue origin of suspicious body fluid stains found at crime scenes and determine whether the body fluid stains at the crime scene are one or several types among saliva, semen, and vaginal secretions.Methods Thirty saliva samples, forty semen samples, and forty vaginal secretion samples (half from 2015 and half from 2024) were collected from healthy adult volunteers. Through primer designing, system formulation, and PCR condition optimization, a multiplex fluorescent amplification system was constructed. The specificity, sensitivity, and detection ability for mixed samples of this system were investigated, and it was tested using real crime scene materials. In the primer design stage, to reduce the requirements for RNA template quality, the amplification products were set within 80-300 bp. In the system formulation stage, dominant and subordinate primers were mainly considered. By reducing the concentration of dominant primers and increasing that of subordinate primers, a capillary electrophoresis spectrum with an appropriate peak height ratio was finally obtained. Additionally, gradient experiments were designed to adjust the concentrations of PCR reagents and PCR amplification conditions, and multiple versions of DNA amplification enzymes were optimized to achieve the best experimental results.Results Through statistical analysis, there was no significant difference in the capillary electrophoresis of the 3 types of body fluid samples from the two years (2015 and 2024), demonstrating that the sample preservation method in this study can preserve samples for a relatively long time. The composite amplification system constructed in this study exhibited high specificity for all 3 types of body fluid, with no cross-reactions between the markers of each type of body fluid. The minimum detection thresholds for the 3 types of body fluid reached 0.002 9, 0.001 5, and 0.42 mg/L, respectively. This system also had a high degree of discrimination for mixed samples, especially for semen-saliva mixtures, where each body fluid marker could still be successfully detected when the concentration ratio of semen to saliva was 100:1. Meanwhile, in the two actual cases presented in this article, the application of this composite amplification system performed outstandingly.Conclusion The composite amplification detection system constructed in this study can achieve the correct screening of saliva, semen, and vaginal secretions, overcoming the problems such as low specificity and sensitivity of marker tests and unbalanced RFU values of each marker in previous studies. The specificity and sensitivity meet the practical work requirements, and the operation is simple. It provides an analytical and identification method for body fluid stains in actual case and is applicable to the identification of the tissue origin of biological evidence at crime scenes involving sexual assault, indecent assault, and other criminal acts. In the future, more types of body fluid markers will be screened to expand the types of body fluids detected by the system, and body fluid-specific cSNP and cInDel genetic markers will be introduced to infer the sources (individuals and types) of mixed and complex stains more accurately.]]> 2024/11/6 14:47:13 BAI Yi-Fan,ZHAO He-Miao,CHEN Jing,LIU Hong-Di,YANG Rui-Qin and WANG Chong 61true Objective Obesity has been identified as one of the most important risk factors for cognitive dysfunction. Physical exercise can ameliorate learning and memory deficits by reversing synaptic plasticity in the hippocampus and cortex in diseases such as Alzheimer’s disease. In this study, we aimed to determine whether 8 weeks of treadmill exercise could alleviate hippocampus-dependent memory impairment in high-fat diet-induced obese mice and investigate the potential mechanisms involved.Methods A total of sixty 6-week-old male C57BL/6 mice, weighing between 20-30 g, were randomly assigned to 3 distinct groups, each consisting of 20 mice. The groups were designated as follows: control (CON), high-fat diet (HFD), and high-fat diet with exercise (HFD-Ex). Prior to the initiation of the treadmill exercise protocol, the HFD and HFD-Ex groups were fed a high-fat diet (60% fat by kcal) for 20 weeks. The mice in the HFD-Ex group underwent treadmill exercise at a speed of 8 m/min for the first 10 min, followed by 12 m/min for the subsequent 50 min, totally 60 min of exercise at a 0° slope, 5 d per week, for 8 weeks. We employed Y-maze and novel object recognition tests to assess hippocampus-dependent memory and utilized immunofluorescence, Western blot, Golgi staining, and ELISA to analyze axon length, dendritic complexity, number of spines, the expression of c-fos, doublecortin (DCX), postsynaptic density-95 (PSD95), synaptophysin (Syn), interleukin-1β (IL-1β), and the number of major histocompatibility complex II (MHC-II) positive cells.Results Mice with high-fat diet-induced obesity exhibit hippocampus-dependent memory impairment, and treadmill exercise can prevent memory decline in these mice. The expression of DCX was significantly decreased in the HFD-induced obese mice compared to the control group (P<0.001). Treadmill exercise increased the expression of c-fos (P<0.001) and DCX (P=0.001) in the hippocampus of the HFD-induced obese mice. The axon length (P<0.001), dendritic complexity (P<0.001), the number of spines (P<0.001) and the expression of PSD95 (P<0.001) in the hippocampus were significantly decreased in the HFD-induced obese mice compared to the control group. Treadmill exercise increased the axon length (P=0.002), dendritic complexity (P<0.001), the number of spines (P<0.001) and the expression of PSD95 (P=0.001) of the hippocampus in the HFD-induced obese mice. Our study found a significant increase in MHC-II positive cells (P<0.001) and the concentration of IL-1β (P<0.001) in the hippocampus of HFD-induced obese mice compared to the control group. Treadmill exercise was found to reduce the number of MHC-II positive cells (P<0.001) and the concentration of IL-1β (P<0.001) in the hippocampus of obese mice induced by a HFD.Conclusion Treadmill exercise led to enhanced neurogenesis and neuroplasticity by increasing the axon length, dendritic complexity, dendritic spine numbers, and the expression of PSD95 and DCX, decreasing the number of MHC-II positive cells and neuroinflammation in HFD-induced obese mice. Therefore, we speculate that exercise may serve as a non-pharmacologic method that protects against HFD-induced hippocampus-dependent memory dysfunction by enhancing neuroplasticity and neurogenesis in the hippocampus of obese mice.]]> 2024/11/24 21:06:35 YAN Meng-Si,SHU Lin-Jie,WANG Chao-Ge,CHENG Ran,MU Lian-Wei and LIAO Jing-Wen 60true Objective To investigate the influence of histone deacetylase 3 (HDAC3) on the occurrence, development of psoriasis-like inflammation in mice, and the relative immune mechanisms.Methods Healthy C57BL/6 mice aged 6-8 weeks were selected and randomly divided into 3 groups: control group (Control), psoriasis model group (IMQ), and HDAC3 inhibitor RGFP966-treated psoriasis model group (IMQ+RGFP966). One day prior to the experiment, the back hair of the mice was shaved. After a one-day stabilization period, the mice in Control group was treated with an equal amount of vaseline, while the mice in IMQ group was treated with imiquimod (62.5 mg/d) applied topically on the back to establish a psoriasis-like inflammation model. The mice in IMQ+RGFP966 group received intervention with a high dose of the HDAC3-selective inhibitor RGFP966 (30 mg/kg) based on the psoriasis-like model. All groups were treated continuously for 5 d, during which psoriasis-like inflammation symptoms (scaling, erythema, skin thickness), body weight, and mental status were observed and recorded, with photographs taken for documentation. After euthanasia, hematoxylin-eosin (HE) staining was used to assess the effect of RGFP966 on the skin tissue structure of the mice, and skin thickness was measured. The mRNA and protein expression levels of HDAC3 in skin tissues were detected using reverse transcription real-time quantitative polymerase chain reaction (RT-qPCR) and Western blot (WB), respectively. Flow cytometry was employed to analyze neutrophils in peripheral blood and lymph nodes, CD4+ T lymphocytes, CD8+ T lymphocytes in peripheral blood, and IL-17A secretion by peripheral blood CD4+ T lymphocytes. Additionally, spleen CD4+ T lymphocyte expression of HDAC3, CCR6, CCR8, and IL-17A secretion levels were analyzed. Immunohistochemistry was used to detect the localization and expression levels of HDAC3, IL-17A, and IL-10 in skin tissues.Results Compared with the Control group, the IMQ group exhibited significant psoriasis-like inflammation, characterized by erythema, scaling, and skin wrinkling. Compared with the IMQ group, RGFP966 exacerbated psoriasis-like inflammatory symptoms, leading to increased hyperkeratosis. The psoriasis area and severity index (PASI) skin symptom scores were higher in the IMQ group than those in the Control group, and the scores were further elevated in the IMQ+RGFP966 group compared to the IMQ group. Skin thickness measurements showed a trend of IMQ+RGFP966>IMQ>Control. The numbers of neutrophils in the blood and lymph nodes increased sequentially in the Control, IMQ, and IMQ+RGFP966 groups, with a similar trend observed for CD4+ and CD8+ T lymphocytes in the blood. In skin tissues, compared with the Control group, the mRNA and protein levels of HDAC3 decreased in the IMQ group, but RGFP966 did not further reduce these expressions. HDAC3 was primarily located in the nucleus. Compared with the Control group, the nuclear HDAC3 content decreased in the skin tissues of the IMQ group, and RGFP966 further reduced nuclear HDAC3. Compared with the Control and IMQ groups, RGFP966 treatment decreased HDAC3 expression in splenic CD4+ and CD8+ T cells. RGFP966 treatment increased the expression of CCR6 and CCR8 in splenic CD4+ T cells and enhanced IL-17A secretion by peripheral blood and splenic CD4+ T lymphocytes. Additionally, compared with the IMQ group, RGFP966 reduced IL-10 protein levels and upregulated IL-17A expression in skin tissues.Conclusion RGFP966 exacerbates psoriatic-like inflammatory responses by inhibiting HDAC3, increasing the secretion of the cytokine IL-17A, and upregulating the expression of chemokines CCR8 and CCR6.]]> 2024/12/31 11:50:28 XU Fan,ZHANF Xin-Rui,XIA Yang-Chen,LI Wen-Ting,CHEN Hao,QIN An-Qi,ZHANG Ai-Hong,ZHU Yi-Ran,TIAN Feng and ZHENG Quan-Hui 59true Objective To characterize transmembrane protein 68 (TMEM68) in an alternative triacylglycerol (TAG) biosynthesis pathway, and determine the interplay between TMEM68 and the canonical TAG synthesis enzyme acyl-CoA:diacylglycerol acyltransferase (DGAT).Methods Effects of exogenous fatty acid and monoacylglycerol on TAG synthesis and lipid droplet (LD) formation in TMEM68 overexpression and knockout cells treated with DGAT inhibitor or not were investigated by comparing LD morphology, Oil Red O staining, and measurement of TAG levels. LDs were stained with fluorescence dye and observed by confocal fluorescence microscopy. TAG levels were determined with an enzyme-based triglyceride assay kit. Colocalization of TMEM68 and DGAT1 was detected by co-expression and confocal fluorescence microscopy and their interaction was determined by co-immunoprecipitation. RT-qPCR and immunoblotting assay were used to detect the expression of DGAT1.Results The synthesis of TAG catalyzed by TMEM68 was independent of DGAT activity. Surplus exogenous fatty acids and monoacylglycerol promoted TAG synthesis mainly through DGAT in human neuroblastoma cells. The LDs formed by TMEM68 were different in morphology from those by DGAT. In addition, TMEM68 and DGAT1 colocalized in the same endoplasmic reticulum (ER) compartment but did not interact physically. TMEM68 overexpression reduced the expression of DGAT1, the major DGAT enzyme involved in TAG synthesis, while TMEM68 knockout had little impact.Conclusion The TMEM68-mediated TAG synthesis pathway has distinct features from the canonical DGAT pathway, however, TMEM68 and DGAT may coregulate intracellular TAG levels.]]> 2024/11/6 14:54:16 YU Qing,FU Yang-Yang,ZENG Fan-Si,PANG Hui-Min,HUANG Fei-Fei and CHANG Ping-An 58true Objective The aim of this study was to investigate the effect and mechanism of quercetin on lipid droplet formation in foam cells induced by oxidized low-density lipoprotein (ox-LDL).Methods Mouse RAW264.7 cells were induced by 50 mg/L ox-LDL to construct a foam cell model. After different quercetin concentrations were treated for different time, the optimal quercetin concentration and time were screened by CCK8 assay. Based on the constructed foam cell model, the formation of fat droplets was observed by oil red O staining after quercetin treatment with or without AS1842856 (FOXO1 inhibitor). Apoptosis was detected by flow cytometry. The protein expression of FOXO1 in each group was detected by Western blot. Autophagosome formation was observed by acridine orange staining. The mRNA and protein expression levels of Beclin1, LC3II and P62 were detected by qRT-PCR and Western blot.Results After being treated with 100 μmol/L quercetin for 12 h, the formation of fat droplets and apoptosis of foam cells were inhibited (P<0.05). Compared with control group, there was an increase in fat droplet formation and apoptosis (P<0.05), a decrease in autophagosome (P<0.05), a decrease in FOXO1 protein expression (P<0.05), a decrease in Beclin1 and LC3II protein and mRNA expression levels (P<0.05), and the expression levels of P62 protein and mRNA were found to be increased (P<0.05) in model group. Compared with model group, quercetin treatment up-regulated FOXO1 protein expression (P<0.05), induced autophagosome formation (P<0.05), promoted the protein and mRNA expression levels of Beclin1 and LC3II (P<0.05), and inhibited the protein and mRNA expression levels of P62 (P<0.05). In addition, treatment with the FOXO1 inhibitor AS1842856 reversed quercetin’s effect on OX-LDL-induced foam cells.Conclusion Quercetin induced autophagy by upregulating FOXO1 expression and inhibited fat droplet formation induced by OX-LDL.]]> 2024/10/8 21:21:44 ZENG jiang-Qin,SUN Qin-Guo,XU Hong-Jie,DING Xiao-Ming,MOU Yan-Jie and JIANG Yue-Wen 57true Objective Stroke is a leading cause of death and disability worldwide, with ischemic stroke accounting for 80%-85% of cases. Despite the prevalence, effective treatments remain scarce. The compelling evidence suggest that high concentrations of ATP in the brain post-stroke can trigger irreversible neuronal damage and necrosis, contributing to a range of neurocellular dysfunctions. Pyroptosis, a recently identified form of programmed cell death, is characterized by caspase-1 activation and the action of the Gasdermin D (GSDMD) protein family, leading to cell perforation and inflammatory death.Methods In this study, human neuroblastoma SH-SY5Y cells were used to investigate the mechanisms of ATP-induced neurotoxicity and the protective effects of hydrogen sulfide (H2S) against this toxicity through the antagonization of pyroptosis. We employed CCK-8 and LDH assays to assess cell viability. YO-PRO-1 fluorescent dyes and flow cytometry were conducted for detecting changes in cell membrane permeability. Western blot analysis was used to measure protein levels associated with cellular dysfunction.Results Our results indicate that high concentrations of ATP enhance cytotoxicity and increase cell membrane permeability in SH-SY5Y cells, that are mitigated by the H2S donor NaHS. Furthermore, ATP was found to promote the activation of the NOD-like receptor pyrin domain-containing 1 (NLRP-1), caspase-1, and the cleavage of GSDMD, with NaHS significantly attenuating these effects.Conclusion Our research suggests that H2S protects SH-SY5Y cells from ATP-induced neurotoxicity through a mechanism mediated by the NLRP1, caspase-1, and GSDMD pathway.]]> 2024/10/2 0:00:00 REN Yan-Kai,LI Ying-Hong,LI Man-Li,YANG Kun-Li,FAN Zhi-Ru,ZHANG Si-Yu and LI Dong-Liang 56true Objective To investigate whether 2,3,5,4"-tetrahydroxystilbene-2-O-β-glucoside (TSG) ameliorated polycystic ovary syndrome (PCOS)-like characteristics by inhibiting inflammation.Methods PCOS models were established by injecting subcutaneously with dehydroepiandrosterone into female Sprague-Dawley rats, followed by receiving intraperitoneal injection of TSG. The granular cells (GCs) KGN were transfected with small interfering RNAs (si-NC and si-CYP19A1). The cells were preincubated with lipopolysaccharide (LPS) and then treated with or without TSG. The estrous cycle was monitored using vaginal exfoliated cells. The morphology of ovarian follicles was analyzed by H&E staining. ELISA was used to analyze estradiol (E2), testosterone (T), follicle stimulating hormone (FSH), luteinizing hormone (LH), IL-6, TNF-α, AGEs, CRP and Omentin-1 levels in serum. Immunohistochemistry was performed to analyze PCNA and CYP19A1 expressions in the GCs of ovaries. Tunel staining was executed to detect the apoptosis of GCs. Quantitative polymerase chain reaction (qPCR) and Western blot were implemented to measure the expression of CYP19A1 in the ovaries and transfected cells. qPCR was used to analyze the expression of IL-6 and TNF-α in the transfected cells treated with LPS and TSG.Results The estrous cycles were restored in TSG group. Compared with model group, the sinus follicles were reduced and corpus luteums were increased in TSG group. TSG group showed increased E2, and decreased T and LH, compared with model group. Pro-inflammatory factors (IL-6, TNF-α, CRP and AGEs) were decreased, and anti-inflammatory factor (Omentin-1) was increased in TSG group compared with those in model group. TSG could partially inhibit decrease of PNCA-positive GCs and increase of Tunel-positive GCs caused by PCOS. The CYP19A1 expression of GCs in TSG group was upregulated compared with model group. The expressions of IL-6 and TNF-α in si-CYP19A1 cells were increased compared with si-NC cells. Compared with cells (si-NC and si-CYP19A1) treated without LPS, the expressions of IL-6 and TNF-α cells were increased, and the expression of CYP19A1 was downregulated in LPS-preincubated cells. Compared with cells treated with LPS, the expression of IL-6 and TNF-α were decreased, and the expression of CYP19A1 was increased in cells treated with LPS and TSG. Compared with si-NC cells treated with LPS and TSG, the expressions of IL-6 and TNF-α cells were increased in the si-CYP19A1 cells treated with LPS and TSG.Conclusion TSG could alleviate PCOS-like characteristics by increasing the expression of CYP19A1 in GCs to inhibit inflammatory response.]]> 2024/11/19 0:00:00 YAO Jian-Feng,XU Bo,LIU Pin-Yue,CHENG Di,WEI Le,PAN Xin-Yun,MO Zhong-Cheng and LI Ming 55true Objective To explore the mechanism of exercise intervention in improving autism-like behaviors in Shank3 gene knockout (Shank3-/-) induced autism spectrum disorder (ASD) rat models from an autophagy perspective through 8-week swimming intervention.Methods Based on genotype identification and exercise intervention, rats were divided into four groups (n=15): wild-type control group (WC), Shank3-/- control group (KC), wild-type swimming group (WS), and Shank3-/- swimming group (KS). KS and WS groups underwent 8 weeks of swimming exercise, 5 d/week, gradually increasing to and maintaining 40 min/session. Behavioral experiments, including self-grooming test, marble burying test, and hole-board test, were conducted 24 h after the final swimming intervention. Tissue sampling was performed 12 h after behavioral testing. Transmission electron microscopy was used to observe autophagosome numbers in the striatum region. Immunofluorescence staining was employed to observe the expression levels of microtubule-associated protein 1 light chain 3 (LC3) and selective autophagy adaptor protein (p62). Quantitative real-time polymerase chain reaction (qPCR) and Western blot were used to detect protein and mRNA expression of Beclin1, LC3, p62, autophagy-related protein 5 (Atg5), autophagy-related 16-like protein 1 (Atg16L), and lysosome-associated membrane protein 1 (LAMP1) in striatal.Results Compared with the WC group, rats in the KC group exhibited significantly higher self-grooming frequency and duration (P0.05), increased marble burying behavior (P0.01), and elevated frequencies in both hole-board exploration and single-hole exploration (P0.05). Following 8 weeks of swimming intervention, the KS group demonstrated significantly reduced self-grooming duration, marble burying behavior, and single-hole exploration frequency compared to the KC group. Furthermore, compared to the WC group, the KC group displayed abundant autophagosomes in the striatum region, along with significantly elevated protein and mRNA expression levels of Atg5, Atg16L, p62, and LC3II/LC3I ratio (P0.05), increased Beclin1 protein levels (P0.05), and markedly decreased LAMP1 protein and mRNA expression levels (P0.05). Following the 8-week swimming intervention, the KS group exhibited significantly reduced protein and mRNA expression levels of Atg5, Atg16L, p62, and LC3II/LC3I ratio (P0.05), decreased Beclin1 protein levels (P0.05), and significantly elevated LAMP1 protein and mRNA expression levels (P0.05) compared to the KC group.Conclusion Early 8-week swimming can alleviate stereotyped behaviors in Shank3-/- rats by regulating striatal cell autophagy.]]> 2024/10/2 8:03:25 XUE Ya-Qi,LIU Niu,WANG Shi-Jiao,BA Yi and ZHEN Zhi-Ping 54true Objective Electroencephalography (EEG) serves as a non-invasive electrophysiological monitoring technique employed to record brain electrical activity. Nonetheless, traditional EEG electrodes are susceptible to reference activation influences and exhibit limited spatial resolution. Laplacian electrodes, devoid of reference dependencies, possess the potential to amplify the spatial resolution of EEG recordings. Anchored in the utilization of bipolar concentric ring Laplacian electrodes, this study delves into the autonomous referencing attributes intrinsic to Laplacian electrodes. Furthermore, it conducts a comparison of spatial resolution disparities between Laplacian electrodes and their conventional counterparts.Methods A three-dimensional (3D) hemispherical tank experiment was conducted utilizing 21 Ag/AgCl bipolar concentric ring Laplacian electrodes to simulate whole-brain signal acquisitions. A sinusoidal signal with an amplitude of 400 mVpp@13 Hz was employed for detection. The positions of the ground electrodes in the Laplacian electrode array were varied, alongside the reference electrode positions in the case of the traditional electrodes. Subsequently, the spatial distribution of the 13 Hz source frequency component was extracted and subjected to comprehensive analysis.Results With varying ground electrode positions, the spatial distribution of the signal-to-noise ratio (SNR) among Laplacian electrodes maintains remarkable consistency, yielding a correlation coefficient of 0.94. In contrast, for traditional electrodes, the correlation coefficient for SNR distribution under distinct reference electrode positions barely reaches 0.07. While Laplacian electrodes exhibit independence from reference electrodes, traditional counterparts display a notable susceptibility to changes in reference electrode positions. Comparing amplitude’s 3 dB attenuation area ratio, Laplacian electrodes showcase a mere 2.1% reduction, a significantly favorable outcome when juxtaposed with the 6.9% reduction evident in traditional electrodes. Similarly, the SNR’s 3 dB attenuation area ratio for Laplacian electrodes is a mere 1.0%, contrasting with the considerably higher figure of 30.1% for traditional electrodes.Conclusion Laplacian electrodes remain impervious to reference electrode influence, displaying distinctive reference-independent attributes, in addition to boasting a heightened spatial resolution. These characteristics imbue them with the capacity to achieve heightened precision in localizing brain electrical activities, thus constituting a cornerstone for the integration of Laplacian electrodes into brain-computer interfaces (BCIs).]]> 2024/10/5 22:13:45 ZHENG Chun-Hou,HE Feng,SHI Tian-Ning,XUE Jia-Xing,YING Bing-Jie and XU Min-Peng 53true Objective Cancer is a global public health issue that has attracted much attention. Detecting and treating cancer at an earlier time point is the key to improving cancer survival rates. However, due to factors such as high equipment cost, slow detection speed, and poor detection accuracy, the promotion of early cancer screening is limited. Therefore, this paper proposes a high-precision and high-speed bioimpedance spectroscopy detection method for tumor identification based on multi-frequency synchronous bioimpedance spectroscopy technology.Methods First, based on the multi-frequency synchronization technology, this paper built a multi-frequency synchronous bioimpedance spectrum detection system, realized the high-speed detection of bioimpedance spectrum, designed concentric circle sensors to reduce the influence of biological tissue anisotropy on impedance detection, and improved the discrimination of bioimpedance spectrum between different tissues. Secondly, a gastric wall tissue model was established, and the degree of anisotropy influence on traditional four-electrode sensors and concentric circle sensors was studied through simulation. Finally, through pork tissue detection experiments and clinical gastric cancer tissue detection experiments, it was verified that the multi-frequency synchronous bioimpedance spectroscopy detection system using concentric circle sensors has higher detection accuracy.Results The experimental results show that when using concentric circle sensors, the average overlap rate of detection results is 13.4%, which is 41.7% lower than that of traditional electrodes, and the average discrete coefficient (Cv) is 7.6%, which is 54.0% lower than that of traditional electrodes. The multi-frequency synchronous bioimpedance spectrum detection system takes about 20 ms to perform a detection, and the detection method proposed in this paper has higher detection accuracy and detection speed. Finally, the concentric circle electrodes were selected to conduct clinical experiments on human gastric cancer tissue, and normal tissue and tumor tissue were successfully distinguished.Conclusion The high-precision and high-speed bioimpedance spectroscopy detection method for tumor identification proposed in this paper can effectively reduce the influence of anisotropy of biological tissues and obtain higher-precision and higher-speed detection results.]]> 2024/10/18 14:55:14 WANG Zhong-Wei,LIU Kai,HU Song-Pei,ZOU Bin,PAN Min-Hong and YAO Jia-Feng 52true Objective For prepubertal and urgently treated malignant tumor patients, ovarian tissue cryopreservation and transplantation represent more appropriate fertility preservation methods. Current clinical practices often involve freezing ovarian tissue with high concentrations of cryoprotectants (CPAs) and thawing with water baths. These processes lead to varying degrees of toxicity and devitrification damage to ovarian tissue. Therefore, this paper proposes optimized methods for vitrification of ovarian tissues based on sodium alginate hydrogel encapsulation and magnetic induction nanowarming technology.Methods Firstly, the study investigated the effects of sodium alginate concentration, the sequence of hydrogel encapsulation and CPAs loading on vitrification efficiency of encapsulated ovarian tissue. Additionally, the capability of sodium alginate hydrogel encapsulation to reduce the required concentration of CPAs was validated. Secondly, a platform combining water bath and magnetic induction nanowarming was established to rewarm ovarian tissue under various concentrations of magnetic nanoparticles and magnetic field strengths. The post-warming follicle survival rate, antioxidant capacity, and ovarian tissue integrity were evaluated to assess the efficacy of the method.Results The study found that ovarian tissue encapsulated with 2% sodium alginate hydrogel exhibited the highest follicle survival rate after vitrification. The method of loading CPAs prior to encapsulation proved more suitable for ovarian tissue cryopreservation, effectively reducing the required concentration of CPAs by 50%. A combination of 8 g/L Fe3O4 nanoparticles and an alternating magnetic field of 300 Gs showed optimal warming effectiveness for ovarian tissue. Combining water bath rewarming with magnetic induction nanowarming yielded the highest follicle survival rate, enhanced antioxidant capacity, and preserved tissue morphology.Conclusion Sodium alginate hydrogel encapsulation of ovarian tissue reduces the concentration of CPAs required during the freezing process. The combination of magnetic induction nanowarming with water bath provides an efficient method ovarian tissue rewarming. This study offers novel approaches to optimize ovarian tissues vitrification.]]> 2024/8/27 13:39:37 CAO Yu-Kun,YE Na,LI Zheng and ZHOU Xin-Li 51true Objective Chemotherapy is one of the important therapeutic approaches for cancer treatment. However, the emergence of multidrug resistance and side effects significantly limit its application. To address these challenges, chemotherapy is often combined with other drugs or therapies. Among the 13 human fucosyltransferases (FUTs) identified, FUT8 (alpha-(1,6)-fucosyltransferase) is the only enzyme responsible for core fucosylation. Core fucosylation plays an important role in cancer occurrence, metastasis and chemotherapy resistance, making the suppression of FUT8 a potential strategy for reversing multidrug resistance. This study aims to evaluate the feasibility of combining the small molecule FUT8 inhibitor 2FF (2-deoxy-2-fluoro-L-fucose) with the clinical chemotherapeutic drug doxorubicin (DOX) for treating malignant tumors.Methods The human hepatocellular carcinoma cell line HepG2 and mouse colon cancer cell line CT26 cells were treated with 2FF, DOX or their combination and core fucosylation levels were assessed using Lectin Blot. HepG2 and CT26 cells were exposed to 50 μmol/L 2FF for 72 h, followed by treatment with a gradient concentration of DOX for 24 h. Cell viability and IC50 values were determined via the CCK-8 assay. Transwell invasion assays were conducted to evaluate the combited effect of 2FF and DOX on the invasion ability of HepG2 cells. Flow cytometry was performed to analyze the impact of 2FF, DOX and their combination on membrane PD-L1 expression of HepG2 cells. To assess the in vivo effect, 6 to 8 week old female BALB/c mice (20-25 g), were subcutaneously injected with 1×106 CT26 cells into the right axilla (four groups, six mice in each group). After the average tumor volume reached 100 mm3, mice were treated with DOX, 2FF, their combination, or saline (mock group) every other day. DOX was administanted intraperitoneally (2 mg/kg), 2FF intravenously (5 mg/kg), and the combination group, received the both treatment. Tumor size was measured every other day using a vernier caliper.Results This study demonstrated that DOX upregulates the core fucosylation level in HepG2 and CT26 cells,while 2FF effectively inhibits this DOX-induced effect. Furthermone, 2FF enhanced the sensitivity of HepG2 and CT26 cells to DOX. The combination of 2FF and DOX synergistically inhibited the invasion ability of HepG2 cells, and enhanced the anti-tumor efficacy of CT26 subcutaneous tumor model in BALB/c mice. However the combination thertment led to weight loss in mice. In addition, DOX increased the cell surface PD-L1 expression in HepG2 cells, which was effectively suppressed by 2FF.Conclusion The FUT8 inhibitor 2FF effectively suppresses DOX-induced upregulation of core fucosylation and PD-L1 levels in tumor cells, and 2FF synergistically enhances the anticancer efficacy of DOX.]]> 2024/9/4 16:03:12 XIE Zhi-Dong and ZHANG Xiao-Lian 50true Objective At present, the most commonly used photosensitizers in photodynamic therapy are still chemical photosensitizers, such as porphyrin and methylene blue, in order to specifically target cellular tissues, and thus poison cells, chemical photosensitizers need to use antibody conjugation or a transgenically encoded tag with affinity for the modified photosensitizing ligand, e.g. FlAsH, ReAsh or Halo Tag. Gene-encoded photosensitizers can directly poison cells by targeting specific cell compartments or organelles. However, currently developed gene-encoded photosensitizers have low reactive oxygen species production and low cytotoxicity, so it is necessary to continue to develop and obtain photosensitizers with higher reactive oxygen species production for the treatment of microbial infections and tumors.Methods In this study, we developed a photosensitizer LovPSO2 based on the light-oxygen-voltage (LOV) structural domain of phototropin-1B-like from Oryza sativa japonica. LovPSO2 was expressed in E. coli BL21(DE3) and purified to obtain protein samples, the purified protein samples were added 3 μmol/L singlet oxygen probe of SOSG and 5 μmol/L superoxide anion probe of DHE after fixed to A445=0.063±0.003, respectively, then measured every 2 min of singlet oxygen production for 10 min and every 1 min of superoxide anion production for 5 min under blue light irradiation at 445 nm, 70 μmol·m-2·s-1.Results The results showed that LovPSO2 could produce a large amount of singlet oxygen under blue light irradiation at 445 nm, 70 μmol·m-2·s-1, and its singlet oxygen quantum yield was 0.61, but its superoxide anion yield was low, so in order to improve the superoxide anion yield of LovPSO2, a mutant with a relatively high superoxide anion yield was obtained by further development and design on its basis LovPRO2. The stability of proteins is crucial for research in drug development and drug delivery, among others. Temperature and light are the key factors affecting the production of reactive oxygen species (ROS) by photosensitive proteins and their stability, while the temperature in cell culture and mammals in vivo is about 37°C, and the temperature inside tumor cells is about 42-45°C. Therefore, we further analyzed the photostability of miniSOG, SOPP3, LovPSO2, and LovPRO2 and their thermostability at 37℃ and 45℃. The analysis of proteins thermostability showed that LovPSO2 and LovPRO2 had better thermostability at 37℃ and 45℃, respectively. Analysis of the photostability of the proteins showed that LovPRO2 had better photostability. In addition, to further determine the phototoxic effects of photosensitizers, LovPSO2 and LovPRO2 were expressed in E. coli BL21(DE3) and HeLa cells, respectively. The results showed that LovPSO2 and LovPRO2 had better phototoxicity to E. coli BL21(DE3) under blue light irradiation, and the cellular phototoxicity lethality was as high as 90% after 30 min of continuous light irradiation, but the phototoxicity was weaker in HeLa cells. The reason for this result may be that the intracellular environment exacerbated the photobleaching of FMN encapsulated by LovPSO2 and LovPRO2, respectively, which attenuated the damage of reactive oxygen species to animal cellular tissues, limiting its use as a mechanistic tool to study oxidative stress.Conclusion LovPSO2 and LovPRO2 can be used as antibacterial photosensitizers, which have broader application prospects in the food and medical fields.]]> 2024/9/10 0:00:00 XU Shuang,WAN Ben,SHA Na and ZHAO Kai-Hong 49true Objective To investigate the role of paraventricular nucleus (PVN) corticotropin releasing hormone (CRH) neurons in chronic restraint stress (CRS)-induced anxiety-like behavior. And whether exercise relieves chronic restraint stress-induced anxiety through PVN CRH neurons.Methods Twenty 8-week-old male C57BL/6J mice were randomly divided into control (Ctrl) group and chronic restraint stress (CRS) group. The open field test (OFT) and elevated plus maze (EPM) were used to evaluate anxiety-like behavior of the mice. Food intake was recorded after CRS. Immunofluorescence staining was used to label the expression of c-Fos expression in PVN and calculate the co-expression of c-Fos and CRH neurons. We used chemogenetic activation of PVN CRH neurons to observed the anxiety-like behavior. 8-week treadmill training (10-16 m/min, 60 min/d, 6 d/week) were used to explore the role of exercise in ameliorating CRS-induced anxiety behavior and how PVN CRH neurons involved in it.Results Compared with Ctrl group, CRS group exhibited significant anxiety-like behavior. In OFT, the mice in CRS groups spent less time in center area (P<0.001). In EPM, the time in open arm in CRS group were significantly decreased (P<0.001). Besides, food intake was also suppressed in CRS group compared with Ctrl group (P<0.05). Compared with Ctrl group, CRS significantly increase c-Fos expression in PVN and most of CRH neurons co-express c-Fos (P<0.001). Chemogenetic activation of PVN CRH neurons induced anxiety-like behavior (P<0.05) and inhibited feeding behavior (P<0.01). Exercise relieves chronic restraint stress-induced anxiety (P<0.001) and relieved the anorexia caused by chronic restraint stress (P<0.05). Aerobic exercise inhibited the CRS labeled c-Fos in PVN CRH neurons (P<0.001). Furthermore, ablation of PVN CRH neurons attenuated CRS induced anxiety-like behavior.Conclusion CRS activated PVN CRH neurons, induced anxiety-like behavior and reduced food intake. 8-week exercise attenuated CRS-induced anxiety-like behavior through inhibiting PVN CRH neuron. Ablation of CRH PVN neurons ameliorated CRS-induced anxiety-like behavior. These finding reveals a potential neural mechanism of exercise-relieving CRS-induced anxiety-like behavior. This provides a new idea and theoretical basis for the treatment of anxiety and related mental disorders.]]> 2024/9/5 16:38:29 CHEN Jing, CHEN Cong-Cong, ZHANG Kai-Na, LAI Yu-Lin, ZOU Yang 48true Objective Tomatoes are one of the highest-yielding and most widely cultivated economic crops globally, playing a crucial role in agricultural production and providing significant economic benefits to farmers and related industries. However, early blight in tomatoes is known for its rapid infection, widespread transmission, and severe destructiveness, which significantly impacts both the yield and quality of tomatoes, leading to substantial economic losses for farmers. Therefore, accurately identifying early symptoms of tomato early blight is essential for the scientific prevention and control of this disease. Additionally, visualizing affected areas can provide precise guidance for farmers, effectively reducing economic losses. This study combines hyperspectral imaging technology with machine learning algorithms to develop a model for the early identification of symptoms of tomato early blight, facilitating early detection of the disease and visual localization of affected areas.Methods To address noise interference present in hyperspectral images, robust principal component analysis (RPCA) is employed for effective denoising, enhancing the accuracy of subsequent analyses. To avoid insufficient information representation caused by the subjective selection of regions of interest, the Otsu’s thresholding method is utilized to extract tomato leaves effectively from the background, with the average spectrum of the entire leaf taken as the primary object of study. Furthermore, a comprehensive spectral preprocessing workflow is established by integrating multivariate scatter correction (MSC) and standardization methods, ensuring the reliability and effectiveness of the data. Based on the processed spectral data, a discriminant model utilizing a linear kernel function support vector machine (SVM) is constructed, focusing on characteristic wavelengths to improve the model"s discriminative capability.Results Compared to full-spectrum modeling, this approach results in an 8.33% increase in accuracy on the test set. After optimizing the parameters of the SVM model, when C=1.64, the accuracies of the training set and test set reach 91.67% and 94.44%, respectively, demonstrating a 1.19% increase in training set accuracy compared to the unoptimized model, while maintaining the same accuracy on the test set, effectively alleviating issues of underfitting.Conclusion This study successfully establishes an early discriminant model for tomato early blight using hyperspectral imaging and achieves visualization of early symptoms. Experimental results indicate that the SVM discriminant model based on characteristic wavelengths and a linear kernel function can effectively identify early symptoms of tomato early blight. Visualization of these symptoms in terms of disease probability allows for a more intuitive detection of early diseases and timely implementation of corresponding control measures. This visual analysis not only enhances the efficiency of disease identification but also provides farmers with more straightforward and practical information, aiding them in formulating more reasonable prevention strategies. These research findings provide valuable references for the early identification and visualization of plant diseases, holding significant practical implications for monitoring, identifying, and scientifically preventing crop diseases. Future research could further explore how to apply this model to disease detection in other crops and how to integrate IoT technology to create intelligent disease monitoring systems, enhancing the scientific and efficient management of crops.]]> 2024/10/2 9:50:33 BAO Hao,HUANG Li,ZHANG Yan and PANG Hao 47true Objective The controllability changes of structural brain network were explored based on the control and brain network theory in young smokers, this may reveal that the controllability indicators can serve as a powerful factor to predict the sleep status in young smokers.Methods Fifty young smokers and 51 healthy controls from Inner Mongolia University of Science and Technology were enrolled. Diffusion tensor imaging (DTI) was used to construct structural brain network based on fractional anisotropy (FA) weight matrix. According to the control and brain network theory, the average controllability and the modal controllability were calculated. Two-sample t-test was used to compare the differences between the groups and Pearson correlation analysis to examine the correlation between significant average controllability and modal controllability with Fagerstr?m Test of Nicotine Dependence (FTND) in young smokers. The nodes with the controllability score in the top 10% were selected as the super-controllers. Finally, we used BP neural network to predict the Pittsburgh Sleep Quality Index (PSQI) in young smokers.Results The average controllability of dorsolateral superior frontal gyrus, supplementary motor area, lenticular nucleus putamen, and lenticular nucleus pallidum, and the modal controllability of orbital inferior frontal gyrus in the young smokers’ group, supplementary motor area, gyrus rectus, and posterior cingulate gyrus, were all significantly different from those of the healthy controls group (P<0.05). The average controllability of the right supplementary motor area (SMA.R) in the young smokers group was positively correlated with FTND (r=0.393 0, P=0.004 8), while modal controllability was negatively correlated with FTND (r=-0.330 1, P=0.019 2).Conclusion The controllability of structural brain network in young smokers is abnormal. which may serve as an indicator to predict sleep condition. It may provide the imaging evidence for evaluating the cognitive function impairment in young smokers.]]> 2024/6/18 0:00:00 DING Jing-Jing,DONG Fang,WANG Hong-De,YUAN Kai,CHENG Yong-Xin,WANG Juan,MA Yu-Xin,XUE Ting and YU Da-Hua 46true Objective INF2 is a member of the formins family. Abnormal expression and regulation of INF2 have been associated with the progression of various tumors, but the expression and role of INF2 in hepatocellular carcinoma (HCC) remain unclear. HCC is a highly lethal malignant tumor. Given the limitations of traditional treatments, this study explored the expression level, clinical value and potential mechanism of INF2 in HCC in order to seek new therapeutic targets.Methods In this study, we used public databases to analyze the expression of INF2 in pan-cancer and HCC, as well as the impact of INF2 expression levels on HCC prognosis. Quantitative real time polymerase chain reaction (RT-qPCR), Western blot, and immunohistochemistry were used to detect the expression level of INF2 in liver cancer cells and human HCC tissues. The correlation between INF2 expression and clinical pathological features was analyzed using public databases and clinical data of human HCC samples. Subsequently, the effects of INF2 expression on the biological function and Drp1 phosphorylation of liver cancer cells were elucidated through in vitro and in vivo experiments. Finally, the predictive value and potential mechanism of INF2 in HCC were further analyzed through database and immunohistochemical experiments.Results INF2 is aberrantly high expression in HCC samples and the high expression of INF2 is correlated with overall survival, liver cirrhosis and pathological differentiation of HCC patients. The expression level of INF2 has certain diagnostic value in predicting the prognosis and pathological differentiation of HCC. In vivo and in vitro HCC models, upregulated expression of INF2 triggers the proliferation and migration of the HCC cell, while knockdown of INF2 could counteract this effect. INF2 in liver cancer cells may affect mitochondrial division by inducing Drp1 phosphorylation and mediate immune escape by up-regulating PD-L1 expression, thus promoting tumor progression.Conclusion INF2 is highly expressed in HCC and is associated with poor prognosis. High expression of INF2 may promote HCC progression by inducing Drp1 phosphorylation and up-regulation of PD-L1 expression, and targeting INF2 may be beneficial for HCC patients with high expression of INF2.]]> 2024/8/31 22:33:11 WANG Hai-Biao,LIN Man,YE Fu-Sang,SHI Jia-Xin,LI Hong,YE Meng and WANG Jie 45true Objective Ulcerative colitis is a prevalent immunoinflammatory disease. Th17/Treg cell imbalance and gut microbiota dysregulation are key factors in ulcerative colitis pathogenesis. The actin cytoskeleton contributes to regulating the proliferation, differentiation, and migration of Th17 and Treg cells. Wdr63, a gene containing the WD repeat domain, participates in the structure and functional modulation of actin cytoskeleton. Recent research indicates that WDR63 may serve as a regulator of cell migration and metastasis via actin polymerization inhibition. This article aims to explore the effect of Wdr63 deletion on Th17/Treg cells and ulcerative colitis.Methods We constructed Wdr63-/- mice, induced colitis in mice using dextran sulfate sodium salt, collected colon tissue for histopathological staining, collected mesenteric lymph nodes for flow cytometry analysis, and collected healthy mouse feces for microbial diversity detection.Results Compared with wild-type colitis mice, Wdr63-/- colitis mice had a more pronounced shortening of colonic tissue, higher scores on disease activity index and histological damage index, Treg cells decreased and Th17 cells increased in colonic tissue and mesenteric lymph nodes, a lower level of anti-inflammatory cytokine IL-10, and a higher level of pro-inflammatory cytokine IL-17A. In addition, WDR63 has shown positive effects on maintaining intestinal microbiota homeostasis. It maintains the balance of Bacteroidota and Firmicutes, promoting the formation of beneficial intestinal bacteria linked to immune inflammation.Conclusion Wdr63 deletion aggravates ulcerative colitis in mice, WDR63 inhibits colonic inflammation likely by regulating Th17/Treg balance and maintains intestinal microbiota homeostasis.]]> 2024/8/24 21:04:40 ZHU Hao,ZHU Meng-Yuan,CAO Yang-Yang,YANG Qiu-Bo and FAN Zhi-Peng 44true Objective Autism spectrum disorder (ASD) is a neurodevelopmental condition characterized by difficulties with communication and social interaction, restricted and repetitive behaviors. Previous studies have indicated that individuals with ASD exhibit early and lifelong attention deficits, which are closely related to the core symptoms of ASD. Basic visual attention processes may provide a critical foundation for their social communication and interaction abilities. Therefore, this study explores the behavior of children with ASD in capturing attention to changes in topological properties.Methods Our study recruited twenty-seven ASD children diagnosed by professional clinicians according to DSM-5 and twenty-eight typically developing (TD) age-matched controls. In an attention capture task, we recorded the saccadic behaviors of children with ASD and TD in response to topological change (TC) and non-topological change (nTC) stimuli. Saccadic reaction time (SRT), visual search time (VS), and first fixation dwell time (FFDT) were used as indicators of attentional bias. Pearson correlation tests between the clinical assessment scales and attentional bias were conducted.Results This study found that TD children had significantly faster SRT (P0.05) and VS (P0.05) for the TC stimuli compared to the nTC stimuli, while the children with ASD did not exhibit significant differences in either measure (P0.05). Additionally, ASD children demonstrated significantly less attention towards the TC targets (measured by FFDT), in comparison to TD children (P0.05). Furthermore, ASD children exhibited a significant negative linear correlation between their attentional bias (measured by VS) and their scores on the compulsive subscale (P0.05).Conclusion The results suggest that children with ASD have difficulty shifting their attention to objects with topological changes during change detection. This atypical attention may affect the child’s cognitive and behavioral development, thereby impacting their social communication and interaction. In sum, our findings indicate that difficulties in attentional capture by TC may be a key feature of ASD.]]> 2024/6/27 11:29:54 XU Hui-Lin,XI Huan-Jun,DUAN Tao,LI Jing,LI Dan-Dan,WANG Kai and ZHU Chun-Yan 43true Objective Early blight is a common destructive disease in the growth process of Solanaceae crops, which can lead to crop failure and serious losses. Traditional crop disease detection methods are difficult to detect disease characteristics in a timely manner during the incubation period of disease, and thus take scientific and effective prevention and control measures. This study obtained hyperspectral images of early blight of peppers at different infection stages through continuous monitoring with a hyperspectral imager. The earliest identifiable time during the incubation period of early blight in peppers (the earliest identifiable time during the incubation period in this experiment was 24 h after inoculation) was determined using the spectral angle cosine-correlation coefficient and Chebyshev distance.Methods Taking the symptoms of the latent period of early blight in peppers as the research object, 13 characteristic wavelengths were selected using a genetic algorithm. An identification model of crop disease latent period symptoms based on spectral features was established through optimized combinations of characteristic wavelengths combined with a logistic regression model. Simultaneously, a recognition model of the latent period of early blight in peppers based on image texture features was established using local binary patterns.Results The experiment was tested with 120 samples. The accuracy of the identification model of crop disease latent period symptoms based on spectral features reached over 93% in both the training set and the test set. The accuracy of the identification model of crop disease latent period symptoms based on texture features reached 98.96% and 100% in the training set and test set, respectively.Conclusion Both spectral features and texture features can be used to detect and identify crop disease latent period symptoms. Texture features more significantly revealed the characteristics of the latent period of the disease compared to spectral features, effectively improving the detection performance of the model. The research results in this article can provide theoretical references for monitoring and identifying other crop disease latent period symptoms.]]> 2024/7/3 14:18:31 SHEN Meng-Jiao,BAO Hao and ZHANG Yan 42true Objective Non-coding RNA (ncRNA) plays a crucial regulatory role in various biological processes. Numerous studies have shown that the functions of ncRNAs depend not only on their nucleotide sequences but also closely on their spatial conformations, particularly their secondary structures. Traditional methods for predicting RNA secondary structures often have low accuracy, usually around 50 to 70 percent, and their performance further declines when dealing with complex structures or long sequences. Although deep learning methods have improved prediction performance to some extent, they still face challenges such as high model complexity, difficulty in capturing long-range dependencies, and poor generalization in predicting complex structures. Therefore, it is necessary for RNA secondary structure prediction to develop new methods.Methods This study proposed an end-to-end prediction model RMDfold, which employed a feature extraction strategy combining residual Mamba (resMamba) and dense connections (Dense). The model framework consisted of three modules: one-dimensional (1D) modeling, feature mapping, and two-dimensional (2D) modeling. For the 1D modeling module, the model learned contextual dependencies among nucleotides from RNA sequences, providing the foundation for possible base-pairing; for the feature mapping module, the 1D features were projected into a 2D space to form a constraint matrix that represented potential base-pairing relationships; for the 2D modeling module, the model further learned the pairing patterns between nucleotides, and determined the unique pairing of each nucleotide through pairing constraints, to obtain the final secondary structure. For both 1D and 2D modeling modules, a four-layer Dense block composed of batch normalization, ReLU activation, and convolution was used to extract short-range features; and a dual-branch residual structure resMamba based on a state-space model was used to model long-range dependencies, thereby achieving effective integration of short- and long-range features. To validate the effectiveness of the proposed method, it was compared with Ufold, REDfold, TransUfold, and sincfold on the three public datasets RNAStralign, ArchiveII, and bpRNA-new.Results The proposed RMDfold method demonstrates superior performance compared with existing algorithms in structure prediction, pseudoknot prediction, sequence prediction across varying lengths, and model complexity analysis, while requiring fewer parameters and achieving faster inference. For the structure prediction, the method achieved F1, Matthews correlation coefficient (MCC), precision and recall of 0.973 5, 0.973 1, 0.975 6 and 0.972 3 on RNAStralign, 0.854 3, 0.855 6, 0.874 7 and 0.876 2 on ArchiveII, and 0.382 8, 0.401 5, 0.536 5 and 0.318 7 on bpRNA-new, respectively. For the pseudoknot prediction based on ArchiveII, the model achieved F1, MCC, precision and recall of 0.741 4, 0.743 3, 0.752 5 and 0.739 6. For the sequence prediction across different lengths, RMDfold maintained an accuracy of 0.70 for sequences ranging from 200 to 500 nt. In terms of model complexity, RMDfold required 2.886 7 M parameters and achieves an inference speed of 0.026 0 s.Conclusion RMDfold enables highly accurate prediction of RNA secondary structures. It helps to deeply and comprehensively reveal the central roles of RNA molecules in gene expression regulation, molecular recognition, and catalysis. and also provides important structure support for elucidating disease-related variant mechanisms, designing RNA-targeted drugs, and advancing research in evolution and comparative genomics.]]> 2025/10/31 12:07:56 HAO Ze-Zhou,YANG Yan-Ling,ZHOU Liang and YAO Xu-Feng 41true Objective This study proposes a fatigue detection method for police extreme training based on electrical impedance imaging technology to prevent muscle damage caused by overstrain during intense physical training.Methods First, based on the mechanism of human anaerobic exercise, lactic acid was identified as a key indicator of muscle fatigue, demonstrating that measuring muscle lactic acid effectively reflects localized fatigue. Second, a numerical simulation model of the human calf was established, and the internal tissue structure of the calf was analyzed to determine the stages of lactic acid diffusion and change. Then, the reconstruction performance of electrical impedance tomography (EIT) in visualizing lactic acid diffusion was compared under three different regularization algorithms, and the most suitable regularization method for subsequent experiments was selected. Finally, a controlled experiment simulating lactate diffusion was conducted to verify the imaging capability of the TK-Noser regularization algorithm in complex imaging fields.Results Simulation results indicate that both the TK-Noser and TV regularization algorithms achieve superior imaging performance, effectively suppressing artifacts in the visualization of lactic acid diffusion inside muscle tissue. The average ICC/RMSE values reached 0.754/0.303 and 0.772/0.320, respectively, while the average SSIM/PSNR values were 0.677/61 dB and 0.488/60 dB, respectively. In the lactate diffusion experiment, the average ICC/SSIM of the EIT reconstruction results based on the TK-Noser regularization algorithm reached 0.701/0.572, respectively. Additionally, compared with the TV regularization algorithm, the TK-Noser algorithm better preserved the shape and structural integrity of the imaging target, with an SSIM value 21.2% higher than that of the TV regularization results. This enhancement ensures the stability of the experimental results and significantly improves the capability of electrical impedance imaging technology in monitoring lactate diffusion within complex fields.Conclusion The proposed method offers real-time convenience and non-invasiveness, making it a promising approach for dynamic monitoring of muscle lactate levels in police officers during extreme physical training.]]> 2025/4/11 8:39:58 LIU Tao,SHI Shu-Sheng,LIU Jun-Feng,LIU Kai and YAO Jia-Feng 40true Objective To address the challenge of identifying unknown animal-derived components in the fields of forensic evidence and food safety, we developed and validated a sensitive and efficient multiplex polymerase chain reaction-capillary electrophoresis (PCR-CE) detection system capable of simultaneously identifying DNA from 13 species—cattle (Bos taurus), mouse (Mus musculus), dog (Canis lupus familiaris), rat (Rattus norvegicus), pig (Sus scrofa), Chinese hamster (Cricetulus griseus), cat (Felis catus), horse (Equus caballus), human (Homo sapiens), chicken (Gallus gallus), duck (Anas platyrhynchos), donkey (Equus asinus), and sheep (Ovis aries)—within a single-tube reaction.Methods Species-specific primers were meticulously designed targeting hypervariable regions of the mitochondrial DNA (including Cytb, COI, 16S rRNA, and ND2 genes), with primer specificity rigorously verified in silico using BLAST analysis against non-target species. A multiplex PCR system was constructed, and critical reaction parameters, including primer concentrations and annealing temperature, were systematically optimized through gradient experiments to ensure balanced amplification of all 13 targets without non-specific products or primer-dimer formation. Amplification products were subsequently separated and detected based on their characteristic fragment lengths and distinct fluorescent labels (6-FAM, HEX) using a capillary electrophoresis platform. The established multiplex PCR-CE system was comprehensively evaluated across several parameters: specificity was tested against 17 species (13 target species and 4 non-target species); sensitivity was determined using serial dilutions of mixed DNA templates; the ability to detect adulteration was assessed using simulated mixed meat samples with known adulteration percentages (10% to 0.1%); and practical applicability was investigated by analyzing 27 commercially available meat products and authentic casework samples from a fatal dog attack incident.Results The optimized multiplex PCR-CE system successfully demonstrated specific and simultaneous amplification for all 13 target species, with zero cross-reactivity observed with 4 non-target species. The system exhibited high sensitivity, with detection limits ranging from 0.05 ng to 0.001 ng of DNA template depending on the species; ten species were detectable at the 0.001 ng level. In simulated adulteration studies, the system reliably detected duck DNA in sheep meat at 0.5%, pork in beef at 0.5%, and horse meat in donkey meat at a remarkably low level of 0.1%. Analysis of commercial meat products revealed an 18.52% (5/27) mislabeling rate. These findings were consistent with validation tests using national and industry standard methods. Furthermore, the system effectively identified human and dog DNA from real forensic case evidence (clothing fragments from bite marks), and even detected trace pig DNA, suggesting the dog’s prior pork consumption.Conclusion This study successfully established a highly specific, sensitive, and practical multiplex PCR-CE system for the simultaneous identification of 13 species. By combining the power of multiplex PCR with capillary electrophoresis, our approach delivers a significant advantage in terms of throughput, resolution, and automation over traditional gel-based methods. The system’s proven effectiveness in detecting low-level adulteration in complex mixtures and processed food products, along with its successful application to genuine forensic specimens, underscores its substantial value and broad potential for routine use in forensic laboratories for evidence analysis and in regulatory settings for ensuring food authenticity and safety monitoring.]]> 2025/10/17 13:11:36 GUO Run-Ze,HU Sheng,CHEN Li-Ping,ZHANG Ying,HE Yong-Feng,SONG Zhen,YANG Rui-Qin and SUN Qi-Fan 39true Objective This study aimed to comprehensively investigate the potential protective effects and underlying mechanisms of taurine against dihydrotestosterone (DHT)-induced androgenetic alopecia (AGA) in male C57BL/6 mice, with a focus on hair follicle cycle modulation, cellular proliferation/apoptosis, and key related signaling pathways.Methods Six-week-old female C57BL/6 mice were initially used to assess the hair growth-promoting potential of taurine. After acclimatization, they were randomly assigned to three groups (n=8): control (regular drinking water), taurine (drinking water containing 1% taurine), and minoxidil (topical 2% minoxidil, positive control). For the AGA study, male C57BL/6 mice were randomly divided into five groups (n=8): control (physiological saline), DHT (model group, 1 mg/d DHT), DHT+low-dose taurine (1 mg/d DHT+2 mg/d taurine), DHT+high-dose taurine (1 mg/d DHT+10 mg/d taurine), and DHT+minoxidil (positive control, 1 mg/d DHT+topical 2% minoxidil). One day before treatment initiation, dorsal hair was shaved with scissors, and residual hair was removed using a depilatory cream. DHT and taurine were administered via daily intraperitoneal injection. Hair regrowth was assessed by photographing the depilated area at regular intervals and quantified using a four-point grading system (0-3). Dorsal skin samples were collected on day 14 for histological analysis (H&E staining), immunofluorescence staining (Ki67 for proliferation, TUNEL for apoptosis), ELISA (DHT quantification), RT-qPCR, and Western blot analysis to evaluate the expression of key genes and proteins (androgen receptor (AR), transforming growth factor (TGF)-β1, TGF-β2, Dickkopf-1 (DKK1)).Results In female mice, taurine supplementation significantly accelerated hair growth, with effects comparable to minoxidil. This was evidenced by an earlier transition from pink (telogen) to black (anagen) skin and increased hair growth scores. Histological analysis showed that taurine increased hair follicle count and dermal thickness. Immunofluorescence confirmed enhanced keratinocyte proliferation in the hair matrix. In the DHT-induced AGA model, DHT significantly extended the telogen phase, inhibited hair growth, increased skin DHT content, and induced hair follicle miniaturization. Taurine treatment, particularly at the high dose, effectively counteracted these effects: it promoted the telogen-to-anagen transition and improved hair growth scores. Histomorphometric analysis showed that taurine significantly restored DHT-induced reductions in dermal thickness, hair follicle count, hair bulb depth, and follicle size. Taurine treatment also reduced apoptosis and promoted the proliferation of hair follicle cells, as demonstrated by Ki67 and TUNEL assays. Crucially, RT-qPCR and Western blot analyses revealed that DHT significantly up-regulated the expression of AR, TGF-β1, TGF-β2, and DKK1 at both mRNA and protein levels in dorsal skin. Taurine administration markedly down-regulated the expression of these pathogenic factors, bringing them closer to the levels observed in the control group.Conclusion Taurine demonstrates significant efficacy in alleviating DHT-induced AGA in male C57BL/6 mice. Its protective effects are mediated through multi-faceted mechanisms. (1) Promoting hair follicle cycle progression: it accelerates the transition from telogen to anagen, counteracting DHT-induced prolongation of the telogen phase. (2) Modulating cellular dynamics: it stimulates the proliferation of hair matrix keratinocytes and reduces DHT-induced apoptosis within hair follicle cells. (3) Suppressing androgen-driven pathogenic pathways: it downregulates the expression of critical molecules in the AGA pathway, including AR, the cytokines TGF-β1 and TGF-β2, and the Wnt pathway inhibitor DKK1. Given its favorable safety profile and multi-targeted action, taurine emerges as a promising novel therapeutic candidate or adjunct for treating AGA. Further investigation into its clinical potential and precise molecular mechanisms is warranted. This study provides a robust preclinical foundation for considering taurine supplementation or topical application in hair loss management strategies.]]> 2025/9/22 8:44:34 WU Jin-Qiang,GUO Guo-Guo,ZHANG Xin-Ting,LIU Jin-Jia,WANG Ji-Xiang,HE Xiao-Yan and WANG Hai-Dong 38true Objective In the clinical diagnosis and grading of brain glioma from histopathological slides, whole-slide cell nucleus density estimation is a critical task. This metric is a key biomarker directly correlated with tumor malignancy, proliferative activity, and patient prognosis, as defined by the World Health Organization (WHO) classification system. Glioma density estimation typically relies heavily on the performance of underlying nucleus segmentation. However, segmentation accuracy is challenged by substantial heterogeneity in nucleus morphology and significant staining variations both across slides and within individual specimens. This variability often causes standard semantic segmentation models to overfit the training data, leading to considerable errors in density estimation. Such inaccuracies can compromise downstream pathological assessments, particularly the subjective and time-consuming manual selection of regions of interest (ROI) for grading. To address these limitations, this study aims to develop a precise and robust whole-slide nucleus density estimation method that enhances model generalization and mitigates overfitting, thereby providing an objective, automated tool for glioma analysis.Methods We propose a systematic three-stage pipeline. (1) Preprocessing: whole-slide images (WSIs) of glioma undergo comprehensive preprocessing, including automated data cleaning to discard blurry or artifact-contaminated patches, data augmentation through geometric transformations (e.g., rotation, flipping) to increase dataset diversity, and color normalization. The latter, based on RGB channel ratios, remaps the color space of all patches to a standardized target, reducing domain shifts caused by staining inconsistencies and improving model robustness. A rigorous semi-automated ground-truth annotation protocol is also implemented, where initial binarization assists annotators in accurately labeling even faint or blurry nuclei, ensuring high-quality training data. (2) Segmentation: using the preprocessed patches, we construct a U-net-based segmentation model that incorporates the DropBlock regularization module—here termed U-net+DropBlock. Unlike standard Dropout, which removes individual neurons, DropBlock eliminates contiguous, spatially correlated regions within feature maps. This structural regularization disrupts undesirable spatial dependencies, forcing the network to learn a more distributed and robust feature representation, thereby reducing overfitting. (3) Quantitative analysis: for each segmented patch, density is computed as the ratio of the total nucleus area to the total patch area—a more robust approach than simple nucleus counting, as it accounts for variations in nucleus size. Patch-wise density values are then assembled into a whole-slide density heatmap, offering an intuitive, global overview of tumor cellularity.Results The U-net+DropBlock model was evaluated both quantitatively and qualitatively against state-of-the-art nucleus segmentation methods, including standard U-net and Hover-net. Quantitatively, our model achieved an F1 score of 90.1%, outperforming U-net and Hover-net, which both scored 87.6%. Qualitative analysis confirmed that our method effectively balances precision and recall, substantially reducing the over-segmentation artifacts common with U-net and the under-segmentation issues observed with Hover-net. This enhanced segmentation quality directly improved the accuracy and reliability of the proposed density estimation approach.Conclusion The proposed whole-slide nucleus density estimation method provides a powerful tool for improving the precision and efficiency of glioma diagnosis. By enabling automated, rapid, and objective analysis of cellular density, it overcomes key limitations of manual pathological review. The generated heatmaps allow pathologists to rapidly identify high-density “hotspots” critical for accurate grading and prognostic evaluation, supporting a more standardized and reproducible ROI selection process. This work lays a solid foundation for developing advanced AI-assisted diagnostic systems, paving the way for more precise, efficient, and reproducible glioma assessments in clinical practice.]]> 2025/9/9 17:39:37 XIA Rui-Chen,YE Chen,ZHAO Lai-Ding,LIU Kai,PAN Min-Hong and YAO Jia-Feng 37true Objective This study aims to develop a microwave-induced thermoacoustic and ultrasound dual-modality microscopy system that integrates the advantages of both imaging techniques to investigate the dielectric properties of biological tissues at a microscopic level.Methods This paper first discusses a method to enhance system resolution by combining short-pulse microwave excitation with high-frequency point-focused ultrasonic transducer detection. A three-dimensional microwave-induced thermoacoustic microscopic imaging system was constructed based on this approach and further developed into a dual-modality system capable of both thermoacoustic and ultrasonic imaging. The image reconstruction and dual-modality image fusion strategies are also described. Subsequently, experiments were conducted in the following sequence: imaging of copper wires to evaluate the system’s spatial resolution along the X/Y/Z axes; imaging of tubes containing 3% and 6% saline solutions and tubes filled with coupling agent/vegetable oil to demonstrate the complementary information provided by the two modalities; imaging of brain tissue and bone-cartilage samples to assess the applicability of the technology; and osteoporosis detection to validate the disease diagnostic capability of the dual-modality system. The microwave-induced thermoacoustic and ultrasound microscopic images of these samples were verified against corresponding photographs or micro-CT images.Results The thermoacoustic and ultrasonic images of the copper wire closely matched the physical photograph. The three-dimensional resolutions of the microwave-induced thermoacoustic and ultrasound imaging systems, as estimated from the copper wire experiment, were 178×178×88 μm3 and 177×177×42 μm3, respectively. These measured values align well with theoretical predictions. The dual-modality imaging system successfully combines dielectric property differences captured by thermoacoustic imaging and acoustic impedance variations captured by ultrasound imaging, thereby providing both functional and structural information of the samples. Specifically, the system distinguished between tubes containing saline solutions of different concentrations and those containing vegetable oil, demonstrating strong spatial consistency with physical photographs. The thermoacoustic image contrast among saline solutions corresponded to theoretical dielectric properties, while the ultrasonic contrast between saline and oil reflected their difference in acoustic impedance. The system identified multiple brain tissue structures, including the cortex, hippocampus, superior colliculus, corpus callosum, cingulate cortex, and striatum. The bimodal imaging approach exhibited superior performance, visualizing tissue structures with greater clarity and detail than either modality alone. The brain tissue images were consistent with physical photographs, tissue dielectric properties, and publicly available anatomical atlases. The bimodal system clearly delineated cartilage and epiphyseal lines via thermoacoustic imaging, while ultrasonic imaging revealed bone structures. Thermoacoustic imaging alone differentiated bone sections between normal and osteoporotic groups; however, incorporating prior skeletal contour information from ultrasound significantly enhanced discriminatory power, resulting in intergroup differences with higher statistical significance. The imaging results of bone samples corresponded well with physical photographs, micro-CT images, and theoretical analyses of dielectric properties for cartilage, normal bone, and osteoporotic bone.Conclusion The microwave-induced thermoacoustic and ultrasound dual-modality microscopy system developed in this study demonstrates potential for microscopic detection of complex biological tissues based on dielectric properties. It is expected to provide a new imaging tool for functional assessment of brain tissue and the skeletal system, as well as for studies on disease pathogenesis.]]> 2025/9/22 17:53:26 CHI Zi-Hui, NIE Yin-Qiang, GUO Xiang-Wen, DU Shuang, FANG Qiu-Chao, WU Dan, JIANG Hua-Bei 36true Objective In nature, objects cast shadows due to illumination, forming the basis for stereoscopic perception. Birds need to adapt to changes in lighting (meaning they can recognize stereoscopic shapes even when shadows look different) to accurately perceive different three-dimensional forms. However, how neurons in the key visual brain area in birds handle these lighting changes remains largely unreported. In this study, pigeons (Columba livia) were used as subjects to investigate how neurons in pigeon’s ventrolateral mesopallium (MVL) represent stereoscopic shapes consistently, regardless of changes in lighting.Methods Visual cognitive training combined with neuronal recording was employed. Pigeons were first trained to discriminate different stereoscopic shapes (concave/convex). We then tested whether and how light luminance angle and surface appearance of the stereoscopic shapes affect their recognition accuracy, and further verify whether the results rely on specify luminance color. Simultaneously, neuronal firing activity of neurons was recorded with multiple electrode array implanted from the MVL during the presentation of difference shapes. The response was finally analyzed how selectively they responded to different stereoscopic shapes and whether their selectivity was affected by the changes of luminance condition (like lighting angle) or surface look. Support vector machine (SVM) models were trained on neuronal population responses recorded under one condition (light luminance angle of 45°) and used to decode responses under other conditions (light luminance angle of 135°, 225°, 315°) to verify the invariance of responses to different luminance conditions.Results Behavioral results from 6 pigeons consistently showed that the pigeons could reliably identify the core 3D shape (over 80% accuracy), and this ability wasn’t affected by changes in light angle or surface appearance. Statistical analysis of 88 recorded neurons from 6 pigeons revealed that 83% (73/88) showed strong selectivity for specific 3D shapes (selectivity index>0.3), and responses to convex shapes were consistently stronger than to concave shapes. These shape-selective responses remained stable across changes in light angle and surface appearance. Neural patterns were consistent under both blue and orange lighting. The decoding accuracy achieves above 70%, suggesting stable responses under different conditions (e.g., different lighting angles or surface appearance).Conclusion Neurons in the pigeon MVL maintain a consistent neural encoding pattern for different stereoscopic shapes, unaffected by illumination or surface appearance. This ensures stable object recognition by pigeons in changing visual environments. Our findings provide new physiological evidence for understanding how birds achieve stable perception (“invariant neural representations”) while coping with variations in the visual field.]]> 2025/9/10 0:00:00 NIU Xiao-Ke,ZHANG Meng-Bo,PENG Yan-Yan,HAN Yong-Hao,WANG Qing-Yu,DENG Yi-Xin and LI Zhi-Hui 35true Objective Recent studies have highlighted the critical role of NUDT19 in the initiation, progression, and prognosis of specific cancer types. However, its involvement in pan-cancer analysis has not been fully characterized. This study aims to systematically explore the expression patterns, clinical significance, and immune-related functions of NUDT19 in various cancer types through multi-omics analysis, further revealing its potential role in cancer, particularly its functional and therapeutic target value in leukemia.Methods To achieve this goal, various bioinformatics approaches were employed to evaluate the expression patterns, clinical significance, and immune-related functions of NUDT19 in tumors and normal tissues. Additionally, we analyzed the mutation characteristics of NUDT19 and its relationship with epigenetic modifications. Using the single-cell analysis tool SingleCellBase, we explored the distribution of NUDT19 across different cell subpopulations in tumors. To validate these findings, qRT-PCR was used to measure NUDT19 expression levels in specific tumor cell lines, and we established acute myeloid leukemia (AML) cell lines (HL-60 and THP-1) to conduct NUDT19 knockdown and overexpression experiments, assessing its effects on leukemia cell proliferation, apoptosis, and invasion.Results Pan-cancer analysis revealed the dysregulated expression of NUDT19 across multiple cancer types, which was closely associated with poor prognosis, clinical staging, and diagnostic markers. Furthermore, NUDT19 was significantly correlated with tumor biomarkers, immune-related genes, and immune cell infiltration in different cancers. Mutation analysis showed that multiple mutations in NUDT19 were significantly associated with epigenetic changes. Single-cell analysis revealed the heterogeneity of NUDT19 expression in cancer cells, suggesting its potentially diverse functional roles in different cell subpopulations. qRT-PCR experiments confirmed the significant upregulation of NUDT19 in various tumor cell lines. In AML cell lines, NUDT19 knockdown led to reduced cell proliferation and invasion, with increased apoptosis, while NUDT19 overexpression significantly enhanced cell proliferation and invasion while reducing apoptosis.Conclusion This study demonstrates the diverse roles of NUDT19 in various cancer types, with a particularly prominent functional role in leukemia. NUDT19 is not only associated with tumor initiation and progression but may also influence cancer progression through the regulation of immune microenvironment and epigenetic mechanisms. Our research highlights the potential of NUDT19 as a therapeutic target, particularly for targeted therapies in malignancies such as leukemia, with significant clinical application prospects.]]> 2025/7/10 11:14:24 LI Xiao-Jin,FENG Shuai,YUAN Zhong-Tao and YANG Tong-Hua 34true Objective Early detection of crop diseases is crucial for effective agricultural management and yield protection. While visible light imaging has been widely applied for disease detection due to its accessibility and non-destructive nature, most existing methods primarily focus on identifying diseases during the symptomatic phase, when visual symptoms are already prominent. However, detecting plant diseases during the incubation period—when symptoms are still subtle or invisible—remains a major challenge due to the lack of distinctive visual cues and limited research methodologies. This study aims to address this gap by proposing a novel three-channel recognition model to accurately identify early blight symptoms during the incubation stage in Solanaceae crops, particularly in chili and tomato, using only visible light images.Methods We established a controlled experimental setup in which healthy leaves and leaves inoculated with early blight pathogens were photographed continuously over time. A total of 1 258 visible light images were collected, capturing various stages of disease progression. From these images, lesion regions were manually annotated. To quantitatively characterize early and subtle color changes within the lesion areas, we extracted color moments—first-order (mean), second-order (standard deviation), and third-order (skewness)—from multiple color spaces, including Lab and HSV. By analyzing the temporal variation of these color moments across disease progression stages, we identified the first-order moment of the saturation (S) channel in the HSV color space as the most sensitive indicator of lesion development on inoculated leaves. Using this insight, we defined four disease categories: healthy, incubation stage, early stage, and late stage. Subsequently, a three-channel classification model was constructed by integrating features from three color channels that provided complementary information. Three-channel models were constructed based on R-G-B, L-a-b, and H-S-V color spaces, respectively, to evaluate performance across different crops and to determine which color representation provides the most discriminative power for identifying disease symptoms during the incubation period.Results The proposed models demonstrated strong classification performance. The three-channel model built using the Lab color space achieved a 94.44% accuracy in recognizing the incubation stage of early blight in pepper, effectively distinguishing subtle pre-symptomatic features from healthy tissue. The model based on the HSV color space achieved 100% accuracy in detecting incubation-stage symptoms in tomato, underscoring the discriminative power of S-channel variations in this context. These results confirm the model’s capability to identify early blight before visible lesions become pronounced, which is essential for timely disease intervention.Conclusion This study presents a new technical pathway for early-stage disease detection using visible light images by focusing on subtle color feature changes during the incubation period. The proposed three-channel recognition model effectively identifies early blight in both chili and tomato, offering a non-destructive, low-cost, and easily deployable solution for early warning and precision agriculture. Furthermore, this framework can be generalized to other crops and diseases where early detection plays a critical role in minimizing yield losses and ensuring sustainable production. The method lays a solid foundation for future research in pre-symptomatic plant disease recognition and provides valuable tools for intelligent crop monitoring and precision management systems.]]> 2025/9/23 0:00:00 PANG Hao and ZHANG Yan 33true Objective The frequency difference limen (FDL) serves as a fundamental metric utilized for effectively quantifying the precise perceptual capabilities of the central auditory system. However, traditional measurement methods rely heavily on the active behavioral responses of subjects and are consequently highly susceptible to the negative influence of confounding subjective factors. Furthermore, existing research paradigms frequently employ uniform stimulus configurations that overlook critical individual perceptual differences. Based on brain-computer interface (BCI) technology, this comprehensive study aims to establish an objective and quantitative evaluation method for auditory frequency discrimination by systematically analyzing and decoding the specific neural responses elicited at the exact threshold state.Methods We designed a personalized rapid serial auditory presentation (RSAP) paradigm customized based on each individual’s precise FDL. A cohort of eleven healthy participants was recruited to evaluate the paradigm using pure-tone sequences at a baseline frequency of 4 000 Hz. This experimental paradigm simulates a realistic auditory perception environment through the continuous presentation of acoustic stimuli, thereby allowing for an in-depth investigation into the specific neural representations evoked by weak frequency deviations at the threshold state. Given that auditory stimulus-evoked response features exhibit complex and differentiated spatiotemporal distribution patterns across multiple frequency domains, this study further deeply integrates the cross-scale feature interaction module with the dynamic spatiotemporal attention allocation strategy, innovatively proposing the Multi-Scale Spatial-Temporal Dual Attention Network (MS-STAMNet). Specifically, the network constructs parallel processing branches with multiple receptive fields and introduces a dynamic adaptive weighting strategy to precisely localize core neural activity signals, further deeply integrating multi-scale information through cross-branch feature information interaction to achieve robust single-trial decoding of weak auditory evoked responses.Results The comprehensive electrophysiological data analysis demonstrated that subtle auditory frequency deviation stimuli presented at the threshold level successfully elicited pronounced N2 and P3 event-related potential features, reflecting pre-attentive mismatch detection and subsequent cognitive evaluation, which were prominently distributed over the frontal, central, and temporal regions of the scalp. In the complex time-frequency domain, the extracted neural response characteristics exhibited distinct, statistically significant event-related synchronization within both the low-frequency δ and θ frequency bands, which was simultaneously accompanied by a widespread, prominent event-related desynchronization within the higher α band. A comparative analysis of model performance demonstrated that MS-STAMNet achieved an average unweighted average recall (UAR) of (69.67±6.12)% and area under the curve (AUC) of 0.761 8±0.07, significantly outperforming the established baseline models such as EEGNet and PLNet. Furthermore, a distinct dissociation phenomenon was verified between neural decoding and behavioral performance through regression analysis (R2=0.016, P=0.709), indicating that this model can effectively capture the implicit features of subtle frequency deviations, even when they fail to trigger explicit conscious responses. Additionally, attention weight visualization analysis further reveals the highly accurate focus of the network on key features concentrated over the bilateral temporal and fronto-parietal regions.Conclusion This study systematically and comprehensively uncovers the multi-dimensional spatiotemporal evolutionary patterns of complex neural responses processing subtle acoustic variations under long-sequence threshold auditory stimulation. Concurrently, it verifies the efficacy and robustness of the proposed MS-STAMNet architecture in accurately deciphering weak, single-trial electroencephalogram signals amidst complex background noise. Ultimately, these neurophysiological and algorithmic findings lay a solid theoretical and methodological foundation for the objective and quantitative evaluation of individual auditory cognitive capabilities in clinical applications, transcending the fundamental limitations of traditional behavioral paradigms and providing robust technical support for future auditory research and related clinical assessments.]]> 2026/7/6 23:17:29 LI Sheng-Ye,XIAO Xiao-Lin,YU Shi-Hang,ZHANG Bei-Bei,AN Xing-Wei,XU Min-Peng and MING Dong 32true Objective Transcranial focused ultrasound (tFUS) has emerged as a promising noninvasive neuromodulation technique capable of modulating neural activity with high spatial specificity. However, whether tFUS stimulation of the primary somatosensory cortex (S1) can reliably evoke conscious tactile perception without external peripheral sensory input remains unclear. Therefore, this study examined the feasibility of inducing hand-related tactile perception through targeted S1 stimulation and further investigated how sonication duration (SD), a key temporal parameter, influences the consistency of this perceptual response. The findings may provide empirical evidence for optimizing tFUS parameters in human somatosensory modulation.Methods Forty-eight healthy adults participated in the study and were assigned to two experiments. Experiment 1 included 24 participants and was designed to preliminarily examine whether S1-targeted tFUS could evoke hand-related tactile sensations. This experiment comprised two sub-experiments, Experiment 1a and Experiment 1b, with 12 participants in each. Specifically, Experiment 1a used a randomized active stimulation protocol targeting the bilateral S1 to assess the initial feasibility of inducing contralateral tactile perception, whereas Experiment 1b employed an alternating hemispheric design incorporating both active and sham stimulation to verify the specificity of the tFUS-induced sensations. Experiment 2 enrolled another 24 participants and used a real-sham dual-probe design to control for spatial tactile cues that might arise from probe-scalp contact. Five SD levels, including 200, 400, 600, 800, and 1 000 ms, were systematically tested to evaluate how stimulation duration influenced the behavioral response. The primary behavioral outcome was the perceptual consistency rate, quantified according to the accuracy of contralateral tactile localization.Results In Experiment 1, 91.7% of participants reported reliable hand-related tactile sensations during active tFUS stimulation, including numbness, tingling, or similar sensory experiences. Contralateral localization accuracy under active stimulation was significantly higher than the 0.50 chance level (P<0.05). In contrast, localization performance under sham stimulation did not differ from chance, suggesting that the observed tactile responses were mainly induced by active S1-targeted tFUS stimulation rather than nonspecific stimulation cues. In Experiment 2, perceptual consistency rates significantly exceeded the chance level under the 400 and 1 000 ms SD conditions (P<0.05), whereas no significant effect was observed at 200, 600, or 800 ms. Pairwise comparisons further showed that the 400 and 1 000 ms conditions produced significantly higher perceptual consistency rates than the 200 ms condition (P<0.05).Conclusion tFUS stimulation of S1 can elicit distinct hand-related tactile sensations without peripheral sensory input, demonstrating the feasibility of using targeted cortical ultrasound stimulation to induce conscious somatosensory perception. The reliability of this perceptual effect was modulated by sonication duration, indicating that temporal stimulation parameters play an important role in shaping both the occurrence and consistency of behavioral responses. Among the tested conditions, 400 ms appeared to provide a favorable temporal window for generating stable tactile perception, whereas excessively short stimulation may be insufficient to produce consistent responses. These findings highlight the sensitivity of somatosensory networks to ultrasonic modulation and provide useful parameter guidance for the application of tFUS in noninvasive sensory enhancement and cortical functional mapping.]]> 2026/6/19 21:26:30 CHEN Xu-Xu,XI Huan-Jun,LIANG Zhen,WANG Bo and ZHAO Xu-Dong 31true Objective Migraine is a leading neurological disorder and the fourth most common cause of years lived with disability worldwide, affecting nearly 116 million individuals. Although pharmacological treatments are available, their efficacy is often limited by side effects and variable response rates. Repetitive transcranial magnetic stimulation (rTMS) over the dorsolateral prefrontal cortex (DLPFC) offers a safe, non-invasive alternative for migraine management. However, the neurophysiological mechanisms, particularly how rTMS modulates local cortical excitability and distributed pain-related circuits, remain poorly understood. Elucidating these mechanisms is essential for optimizing treatment protocols and improving clinical outcomes.Methods This study employed concurrent transcranial magnetic stimulation and electroencephalography (TMS-EEG) to investigate neuroplastic and neurocircuitry mechanisms of DLPFC-rTMS in migraine. Study 1 compared 30 migraineurs and 28 healthy controls to identify abnormalities in TMS-evoked potentials (TEPs) and significant current density (SCD) within sensory-discriminative regions including the primary somatosensory cortex (S1) and posterior insula (pINS), cognitive-affective regions including the anterior insula (aINS) and midcingulate cortex (MCC), and a descending modulatory region, the periaqueductal gray (PAG). Study 2 used a single-blind, crossover, sham-controlled design in 34 healthy participants. Each participant received both active (10 Hz, 80% RMT, 1 500 pulses) and sham DLPFC-rTMS in counterbalanced order. TMS-EEG and cold pain tolerance were assessed before and after each session.Results In Study 1, migraineurs showed a significantly less negative N120 amplitude compared to healthy controls (P=0.027, Cohen’s d=0.60), indicating local intracortical disinhibition. No group differences were observed for N40, P60, or P180 components. At the source level, migraineurs exhibited significantly higher SCD in the S1, pINS, aINS, and MCC (all Q<0.05), but not in the ventroposterior thalamus (vpTHAL), mediodorsal thalamus (mdTHAL), or PAG. In Study 2, active rTMS significantly reduced SCD from pre- to post-stimulation in the S1, aINS, and MCC (all Q<0.05). Sham stimulation also reduced SCD in the S1 (Q<0.05) but not in the aINS or MCC. Although no significant group-level analgesic effect was observed between active and sham conditions (P=0.107), correlation analyses revealed that greater SCD reductions in the S1 and MCC were significantly associated with higher post-rTMS pain tolerance (R=-0.487 and -0.495, both Q<0.01) and larger improvements in pain tolerance (R=-0.487 and -0.451, both Q<0.05). No such correlations were found following sham stimulation, suggesting that the behavioural relevance of neural changes is specific to active rTMS.Conclusion This study provides novel evidence that migraineurs exhibit both local neuroplastic abnormalities (reduced N120 amplitude) and hyperactivity in key pain-processing regions (S1, pINS, aINS, MCC). A single session of DLPFC-rTMS reduced hyperactivity in the aINS, MCC, and S1. Notably, greater reductions in the S1 and MCC were associated with improved pain tolerance. These findings identify distinct cortical circuitries, particularly within the cognitive-affective pain network, that may serve as potential biomarkers for optimizing rTMS treatment in migraine and other chronic pain conditions. Future studies should validate these results in patient populations experiencing spontaneous migraine attacks and explore multi-session or accelerated rTMS protocols.]]> 2026/6/2 15:59:26 JIN Chen-Xia,TAN Bo-Lin,YE Yang,HE Ji-Qing,WANG Ling-Yan,GAO Zhong-Ming,WANG Yu-Jun,LIU Hui-Li,YAN Yong-Xing and CHE Xian-Wei 30true Objective Type I interferon (IFN-I) signaling is essential for antiviral innate immunity, yet its sustained or excessive activation contributes to the pathogenesis of several autoimmune diseases and interferonopathies, such as systemic lupus erythematosus and Aicardi-Goutières syndrome. Current strategies targeting this pathway, exemplified by JAK inhibitors, act mainly on downstream signal transduction and provide limited direct control over upstream IFN-I production, while also carrying the risk of broad immunosuppression. Phyllanthus emblica L. has long been used in traditional medicine for inflammatory disorders, but the bioactive constituent responsible for its regulation of IFN-I signaling and the underlying molecular mechanism have not been clearly defined. This study aimed to identify the active anti-inflammatory component of P. emblica and to characterize its mechanism of action on the IFN-I pathway in macrophages.Methods Active components of P. emblica and their candidate targets were screened by network pharmacology using the TCMSP and DrugBank databases (oral bioavailability≥30%, drug-likeness≥0.18) and intersected with inflammation-related genes retrieved from public databases. The predicted interaction between EGCG and IFN-I pathway proteins (TBK1, IRF3, STAT1) was evaluated by molecular docking, with BX795 and GSK8612 used as reference TBK1 inhibitors. Mechanistic experiments were performed in THP-1-derived macrophages and primary bone marrow-derived macrophages (BMDM). Upstream signaling was activated by transfection of the nucleic acid analogs poly(I∶C) and poly(dA∶dT) or by lipopolysaccharide (LPS) stimulation, whereas downstream signaling was activated by exogenous IFN-β. An siRNA-mediated TREX1 knockdown model was used to mimic endogenous nucleic acid-driven interferonopathy. Expression of IFN-β1 and interferon-stimulated genes (ISGs) was measured by RT-qPCR, protein phosphorylation by Western blot, and IFN-β secretion by ELISA. Cellular thermal shift assay (CETSA) and drug affinity responsive target stability (DARTS) were used to probe the interaction between EGCG and IRF3.Results Network pharmacology identified (-)-epigallocatechin-3-gallate (EGCG) as a candidate IFN-I-suppressive constituent of P. emblica, with predicted binding to TBK1, IRF3, and STAT1. Molecular docking yielded binding energies of -9.2, -7.2, and -8.2 kcal/mol for TBK1, IRF3, and STAT1, respectively, indicating an affinity for TBK1 comparable to that of the reference inhibitors BX795 (-5.7 kcal/mol) and GSK8612 (-6.4 kcal/mol). EGCG suppressed IFN-β1 and ISG mRNA expression under poly (I∶C), poly (dA∶dT), and LPS stimulation in both THP-1 macrophages and BMDM. At the protein level, EGCG reduced the phosphorylation of TBK1 and IRF3 without affecting the levels of the upstream sensors cGAS and RIG-I, and lowered IFN-β secretion in a concentration-dependent manner. CETSA and DARTS showed that EGCG did not enhance the thermal stability or protease resistance of IRF3, indicating that its effect on IRF3 is indirect. Following IFN-β stimulation, prolonged EGCG treatment reduced STAT1 phosphorylation in a time-dependent manner without an apparent change in IRF9, and partially attenuated ISG transcription; this effect was not monotonicly concentration-dependent, and CXCL10 showed the most consistent suppression. In TREX1-knockdown cells, the elevated mRNA levels of ISG15, ISG56, and CXCL10 were reduced by EGCG.Conclusion EGCG suppresses IFN-I responses by concurrently inhibiting TBK1-IRF3-dependent IFN-β production and JAK-STAT1-mediated downstream transcription. These in vitro findings provide a mechanistic basis for the anti-inflammatory use of P. emblica in traditional medicine and identify EGCG as a candidate for further evaluation in interferon-driven autoimmune disease models.]]> 2026/6/18 16:26:03 LI Liang,SHENG Qi-Huan,LIU Huan,YANG Wen-Hao,SHI Jia-Lin,SUN Ying-Jie,JING Rui,MAI Wei-Hua,LI Zhi-Min and XIE Xiao-Li 29true Objective To investigate the novel post-translational modifications (PTMs) of SnRK2.6, a central component in the abscisic acid (ABA) signaling pathway, such as SUMOylation, and to establish a foundation for revealing the physiological functions and molecular mechanisms of SnRK2.6 regulated by these new modifications.Methods The interaction between SnRK2.6 and the SUMO E3 ligase SIZ1, as well as members of the SUMO protease family, was examined using yeast two-hybrid and in vitro pull-down assays. An in vitro SUMOylation system in Escherichia coli was utilized to determine whether SnRK2.6 undergoes SUMOylation. Mass spectrometry, combined with site-directed mutagenesis of candidate lysine residues, was employed to identify potential SUMOylation sites on SnRK2.6. In vitro de-SUMOylation assays were performed to assess whether SUMO proteases interacting with SnRK2.6 could catalyze the removal of SUMO moieties from modified SnRK2.6. The protein stability of SnRK2.6 was assessed in a cell-free degradation assay using bacterial-purified SnRK2.6 incubated with total protein extracts from Col and siz1 mutant seedlings. To dissect the genetic relationship between SnRK2.6 and SIZ1, stomatal aperture assays were performed under ABA treatment using snrk2.6, siz1, and snrk2.6 siz1 double mutant plants.Results SnRK2.6 physically interacts with SIZ1 and the SUMO protease ESD4, with the binding domains localized to the C-terminal region of SIZ1 and the N-terminal region of ESD4, respectively. SnRK2.6 was found to be SUMOylated, exhibiting two distinct high-molecular-mass bands ranging from 70 to 100 ku, indicative of modified forms. Bioinformatics analysis predicted four putative SUMOylation sites on lysine residues K57, K63, K142, and K190. Mass spectrometry identified three SUMOylation sites on K63, K142, and K174. However, individual or combinatorial point mutations on these sites had minimal impact on the pattern or intensity of SUMOylation signals, suggesting that these residues may not be responsible for the SUMOylation on SnRK2.6. Instead, such mutations only weaken the protein stability or accelerate the protein mobility of SnRK2.6. Therefore, the exact SUMOylation sites on SnRK2.6 remain unidentified. In de-SUMOylation experiments, incubation of GST-ESD4 with SUMOylated SnRK2.6 for 1-2 h led to the near-complete disappearance of both SUMOylated bands. In contrast, neither the GST control nor the catalytically inactive mutant GST-ESD4C448S exhibited any de-SUMOylation activity. In protein turnover experiments, SnRK2.6 exhibited markedly enhanced half-life in siz1 compared with Col, indicating that SIZ1-dependent SUMOylation promotes SnRK2.6 turnover. Phenotypically, snrk2.6 mutants were completely insensitive to ABA-induced stomatal closure; siz1 mutants displayed pronounced hypersensitivity; and the snrk2.6 siz1 double mutant phenocopied snrk2.6—showing no significant response to ABA beyond that of the snrk2.6 mutant. These data indicate that SIZ1 acts as a negative regulator of ABA-triggered stomatal closure and SnRK2.6 functions as a positive regulator, and the inhibitory activity of SIZ1 is strictly dependent on SnRK2.6, placing SnRK2.6 genetically upstream of SIZ1 in the ABA signaling pathway.Conclusion SnRK2.6 undergoes SUMOylation, although the specific SUMOylation sites have not been defined. SnRK2.6 is dynamically regulated by reversible SUMOylation—catalyzed by SIZ1 and reversed by ESD4—which controls its protein stability. SUMOylation acts as a destabilizing signal for SnRK2.6, and SIZ1 exerts its negative effect on ABA-triggered stomatal closure probably through promoting SnRK2.6 degradation via SUMOylation. These findings uncover SUMOylation as a critical regulatory layer fine-tuning SnRK2.6 abundance in ABA signaling.]]> 2026/6/3 7:43:15 FU Huan-Huan,WEI Jian,LI Meng-Yao and HAN Yong-Feng 28true Objective Flavonoids are clinically significant natural products, yet their oxygen-glycosylation in aqueous environments relies heavily on expensive nucleotide-activated sugar donors such as UDP-glucose. Liquid-liquid phase separation (LLPS) creates specialized, membraneless physicochemical microenvironments capable of modulating enzymatic functions and overcoming mass transfer limitations. This study aims to investigate whether the gut microbiota-derived DgpB/C complex—a multienzyme system traditionally recognized for cleaving stable C-glycosidic bonds and facilitating isomerization—can undergo functional remodeling within phase-separated condensates. Our core objective is to elucidate the role of phase separation in expanding enzymatic catalytic plasticity and to provide a non-canonical, highly cost-effective biocatalytic mechanism for the direct utilization of free sugars in the synthesis of O-glycosylated natural products.Methods An artificial phase-separation platform was constructed utilizing the multivalent arginine-glycine-glycine motif (RGG)-repeat domain derived from the Caenorhabditis elegans LAF-1 protein. To ensure precise spatial compartmentalization, the DgpB/C complex was specifically recruited into the RGG condensates via a high-affinity SZ1/SZ2 heterodimerization tag system. Condensate formation and substrate partitioning were visualized using light and confocal fluorescence microscopy. The chemical structures and regioselectivity of the reaction products were rigorously characterized using high-performance liquid chromatography (HPLC) and liquid chromatography-mass spectrometry (LC-MS)/MS systems. Furthermore, molecular docking and 20-ns molecular dynamics (MD) simulations were performed via the Hermite platform and Uni-GBSA pipeline to elucidate the structural and thermodynamic basis underlying the phase-transition-induced functional shift.Results We observed that the condensates formed by RGG proteins selectively recruited and significantly enriched hydrophobic flavonoid substrates. Strikingly, within the LLPS microenvironment, the DgpB/C complex—which typically exhibits only degradative or isomerase activities—underwent a profound functional remodeling, transforming into an efficient O-glycosyltransferase. Diverging from canonical pathways that require high-energy donors, the sequestered DgpB/C complex directly utilized unactivated free glucose to catalyze the formation of new O-glycosidic bonds. This remodeled activity was found to be evolutionarily conserved across DgpB/C homologs from diverse gut microbiota strains, such as P581a and W974-1. LC-MS/MS analysis further revealed that the phase-separated environment reduced the regioselectivity constraints of the enzyme, enabling catalytic action on multiple hydroxyl positions of the flavonoid scaffold. MD simulations further indicated that the low-water-activity microenvironment of the condensates reconfigured the conformational dynamics of the catalytic pocket, favoring a spatial orientation highly conducive to dehydration condensation.Conclusion This study demonstrates that LLPS drives the functional remodeling of the gut microbiota enzyme DgpB/C through the reconfiguration of the physicochemical microenvironment. These findings challenge traditional perceptions regarding the functional boundaries of metabolic enzymes and reveal a novel biocatalytic pathway that bypasses the requirement for nucleotide-activated sugars. Consequently, this provides a highly promising artificial compartmentalization strategy for the green manufacturing of complex, high-value-added natural products.]]> 2026/4/15 0:00:00 LI Tian-Yu,LI Ping and MA Wen-Fu 27true Objective This study employs a special conformational engineering (CE) technology to construct an α-chymotrypsin-like active center, which includes a catalytic triad, an oxyanion hole, and a substrate-binding site, on silver nanoparticles (AgNPs), thereby creating an AgNP-based artificial hydrolase with high catalytic activity. This study provides a new approach for the design of highly efficient artificial enzymes and enzyme-mimicking.Methods AgNPs were chosen as the scaffold to build the an α-chymotrypsin-like active center. A special CE procedure enables the designed peptide, Triad5, to adopt an α-helical conformation on AgNPs, with the key catalytic residues located on one side of the α-helix forming a catalytic active center with a catalytic triad, an oxyanion hole, and a substrate-binding site. The CE procedure consists of three steps, including conformation induction via trifluoroethanol (TFE), conformation stabilization on AgNPs via Ag-S bonds, and TFE removal via lyophilization. Circular dichroism (CD) spectra were used to confirm the formation and stabilization of the α-helix conformation. Mutations of the key residues combined with stopped-flow kinetic experiments were used to demonstrate the indispensability of each key residue and the synergistic effects among the catalytic triad, the oxyanion hole, and the substrate-binding site.Results CD spectra show that the designed Triad5 alone is in random coil conformation; when conjugated on AgNPs, Triad5 still remains largely unstructured; but after the CE treatment, Triad5 adopts a typical α-helical conformation on AgNPs as designed, thus produces an AgNP-based artificial hydrolase, Silverzyme. Silverzyme exhibits extremely high hydrolytic activity towards p-nitrophenyl acetate (p-NPA), with an extremely high catalytic turnover number per active site of 3.5 s-1, which is even higher than that of α-chymotrypsin. As a comparison, the AgNP-Triad5 conjugate without CE treatment shows much lower catalytic activity than Silverzyme, highlighting the important role of the right conformation of the active center for the catalytic activity and the power of the CE treatment. When the key residues of the catalytic triad of Silverzyme were mutated to alanine, the overall catalytic efficiency of this mutant dropped by about 2 orders of magnitude, unambiguously demonstrating the key role of the designed catalytic triad. Similarly, when the residues for the oxyanion hole were deleted, the mutant with the intact catalytic triad also showed significantly decreased catalytic activity, highlighting the indispensable role of the oxyanion hole for the catalytic activity. Unexpectedly, when both the catalytic triad and the oxyanion hole were kept intact, a slight change of the binding site also resulted in significantly decreased catalytic activity, indicating that the designed binding site is at the right position to align the substrate in the right orientation in the active center for catalytic hydrolysis. These results confirm the synergy among the catalytic triad, the oxyanion hole, and the substrate-binding site, indicating successful mimicking of the active center of α-chymotrypsin. Moreover, Silverzyme shows better thermal stability than α-chymotrypsin, and can even hydrolyze the tough non-activated ester diethyl phthalate, a priority pollutant by the United States Environmental Protection Agency (USEPA).Conclusion This study successfully mimicked the complex catalytic active center of α-chymotrypsin using a conformational engineering strategy, and produced a highly active artificial hydrolase with a well-defined structure and catalytic mechanism. The findings highlight the significant potential of conformational engineering.]]> 2026/5/23 10:07:21 WANG Yan,ZHOU Tong-Tong,GUO Yuan,WANG Hai-Fang and CAO Ao-Neng 26true Objective Cerebral ischemic injury triggers a complex pathological cascade characterized by excessive reactive oxygen species (ROS) accumulation, persistent oxidative stress, and sustained neuroinflammation in the injured brain microenvironment. These events collectively drive mitochondrial dysfunction, microglial overactivation, pro-inflammatory cytokine release, and progressive neuronal apoptosis, ultimately leading to severe and irreversible neurological deficits. However, conventional therapeutic strategies face critical limitations, including poor blood-brain barrier penetration, insufficient local drug concentration, uncontrolled drug release, and off-target systemic side effects. To address this pathological process, we rationally designed and fabricated an injectable ROS-responsive hydrogel loaded with polydopamine nanoparticles (PDA NPs) for spatiotemporally controlled antioxidation, anti-inflammation, and neuroprotection in the ischemic injury microenvironment. The present study aimed to systematically characterize the physicochemical properties, ROS-responsive drug release behavior, biocompatibility, and neuroprotective efficacy of this composite hydrogel system in vitro.Methods PDA NPs were fabricated via oxidative self-polymerization. The ROS-responsive hydrogel was cross-linked using N1-(4-boronobenzyl)-N3-(4-boronophenyl)-N1,N1,N3,N3-tetramethylpropane-1, 3-diaminium (TSPBA) and polyvinyl alcohol (PVA). Morphology, particle size, Zeta potential, and structure of PDA NPs were characterized by dynamic light scattering (DLS), Zeta potential analysis, scanning electron microscopy (SEM), and transmission electron microscopy (TEM). Microstructure, rheological properties, shear-thinning behavior, and ROS-triggered release profiles of the hydrogel were examined by SEM and rheometry. Biocompatibility was evaluated using HT22 mouse hippocampal neurons with CCK-8 and live/dead staining. An oxygen-glucose deprivation/reoxygenation (OGD/R) model was established to simulate ischemic injury in vitro. ROS levels and neuronal apoptosis were detected by DHE staining and TUNEL assay. Microglial polarization and pro-inflammatory cytokine expression were analyzed using immunofluorescence and RT-qPCR in BV-2 microglia. Transwell co-culture was used to verify the indirect neuroprotection mediated by modulated microglia.Results Characterization results confirmed that the as-prepared PDA NPs were monodispersed spherical nanoparticles with uniform diameter and negative surface potential, demonstrating favorable dispersibility and robust ROS-scavenging activity. The TSPBA-PVA hydrogel exhibited a highly porous interconnected network, suitable mechanical strength, and obvious shear-thinning behavior, supporting its application as an injectable implant. More importantly, the hydrogel displayed typical ROS-responsive degradation and on-demand PDA NP release in a ROS-concentration-dependent manner. In vitro cellular experiments demonstrated that the PDA NP-loaded hydrogel possessed excellent biocompatibility with HT22 cells. In the OGD/R model, the hydrogel significantly reduced intracellular ROS accumulation and markedly suppressed neuronal apoptosis. Furthermore, the composite hydrogel effectively redirected BV-2 microglia from the pro-inflammatory M1 toward the anti-inflammatory M2 phenotypes, downregulated the expression of pro-inflammatory cytokines including TNF-α, IL-1β, and IL-6, and reduced inflammatory damage. Transwell co-culture assays further validated that M2-polarized microglia mediated by the hydrogel significantly enhanced the survival of OGD/R-injured HT22 neurons and attenuated apoptosis.Conclusion In this study, we successfully developed a novel injectable ROS-responsive hydrogel loaded with PDA NPs for synergistic antioxidative and anti-inflammatory neuroprotection. This intelligent hydrogel system enables ROS-triggered on-demand release of PDA NPs, efficiently scavenges excessive ROS, inhibits oxidative stress injury, modulates microglial polarization, and suppresses neuroinflammation, thereby exerting robust neuroprotective effects in vitro. This biomaterial platform provides a promising strategy for the targeted and controlled delivery of bioactive nanomaterials in the central nervous system diseases and establishes a solid experimental foundation for the development of in situ injectable therapies for ischemic brain injury.]]> 2026/5/14 19:41:23 XIAO Yang,LIU Wei,SUN Tian-Yi,SHA Chuan-Lu,WANG Chun-Lan and WANG Chang-Yong 25true Objective Hepatocellular carcinoma (HCC) represents 90% of all primary liver cancers. The main risk factors associated with HCC include viral hepatitis (B and/or C), alcohol abuse, and metabolic dysfunction-associated steatotic liver disease (MASLD), which progressively advance to liver fibrosis, cirrhosis, and ultimately evolve into HCC. Surgical resection represents the most effective treatment for HCC, while recent advances in immunotherapy, including immune checkpoint inhibitors and adoptive cell therapies, have provided improved treatment prospects for patients with unresectable HCC. However, the complex metabolic heterogeneity of HCC limits the therapeutic efficacy. Metabolic intermediates acyl-CoA not only provide energy and substrates for numerous biochemical reactions but also serve as donors for protein lysine acylation, a major class of post-translational modification (PTM). Therefore, a deeper understanding of the molecular mechanisms underlying protein lysine acylation and hepatocarcinogenesis is urgently needed.Methods The levels of protein lysine acylation and silence information regulator 5 (SIRT5) expression levels in clinical HCC samples were analyzed by Western blot. Quantitative malonylome and succinylome of HCC samples were analyzed by antibody-based affinity enrichment coupled with tandem mass spectrometry. The proliferation of HCC cells was analyzed with Cell Counting Kit-8 (CCK-8) assays, the apoptosis was quantified by Annexin V-FITC/propidium iodide (PI) staining coupled with flow cytometry, and the ability of cells to migrate was assayed by Transwell assays. The enzymatic activity of glutathione S-transferase Mu 1 (GSTM1) was quantified. Transgenic mice with hepatic overexpression of SIRT5 were constructed using CRISPR-Cas9, and primary hepatocarcinogenesis was induced by administration of diethylnitrosamine.Results Western blot analysis indicated that the expression level of SIRT5 was elevated in clinical samples from HCC patients, and the levels of lysine malonylation, glutarylation, and succinylation were significantly reduced in HCC tissues. Knockout of SIRT5 in MHCC-97H and MHCC-97L hepatoma cells suppressed cell proliferation, and increased the percentage of apoptotic cells significantly. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses of the differentially malonylome and succinylome of HCC samples revealed significant enrichment in two major classes of biological processes: core energy metabolism (e.g., glycolysis/gluconeogenesis, tricarboxylic acid metabolic process, fatty acid beta oxidation) and detoxification and oxidative stress response (e.g., response to toxic substance, chemical carcinogenesis, reactive oxygen species (ROS)). SIRT5 removes malonylation from lysine residues in GSTM1 and restores its detoxification activity, which is crucial for the survival of hepatocytes under stressed conditions. More importantly, in vivo experiment indicated that hepatic-specific overexpression of SIRT5 in mice accelerated diethylnitrosamine-induced liver fibrosis and hepatocarcinogenesis, indicating the critical role of SIRT5 in HCC progression.Conclusion This study highlights the previously unrecognized SIRT5-GSTM1 axis as a key regulator in hepatocarcinogenesis, and suggests a potential target for the treatment of patients with HCC.]]> 2026/5/9 9:17:09 ZHANG Yu,REN Feng-Rui,LI Jia-Yun,CHEN Xiang-Yu,WANG Zi-Yi,SUN Qi,ZHAO Jun-Cheng,ZHANG Ye,HUANG Zhen,HU Hao,WEI Tao-Tao and XIAO Min 24true Objective As a common lifestyle habit, alcohol consumption has a controversial association with the onset of Parkinson’s disease (PD). To demonstrate the correlation between alcohol consumption and PD and to identify associated genes, we integrated findings from clinical surveys, genomics, transcriptomics, and animal experiments.Methods We investigated the alcohol consumption rates (including both before and after disease onset) among 244 PD patients in China and 177 PD patients from the U.S. NHANES database. Mendelian randomization (MR) analysis was performed using genome-wide association study (GWAS) data for three alcohol-related traits and seven PD-related datasets from the MRC IEU OpenGWAS database. Transcriptomic data from the substantia nigra of PD patients were obtained from three GEO datasets (GSE7621, GSE20141, and GSE49036) to analyze RIT2 gene transcription. Finally, three groups of animal experiments (water/20% ethanol/20% liquor, with 4 C57BL/6J mice per group) were conducted to examine changes in brain RIT2 gene expression and transcriptomic profiles following alcohol consumption.Results The alcohol consumption rates among PD patients in China and the U.S. (9%-18.87%) were significantly lower than the general population rates of 15%-45% in their respective regions (P<0.001), suggesting a possible negative association between alcohol consumption and PD. Subsequently, in 21 bidirectional MR analyses using 3 alcohol-related GWAS datasets and 7 PD-related GWAS datasets, the forward MR analyses (alcohol intake as exposure, PD as outcome) yielded 12 negative associations (ORIVW<1) and 9 positive associations (ORIVW>1). Among these, only two negative associations reached statistical significance: alcohol intake frequency (ORIVW=0.75, 95% CI: 0.60-0.93, P=0.010) and alcohol consumption (ORIVW=0.20, 95% CI: 0.05-0.83, P=0.026). The forward MR analysis (alcohol intake→PD) identified 235 SNPs, annotated to 316 genes, while the reverse MR analyses (PD→alcohol intake) identified 37 SNPs, annotated to 53 genes. Notably, only the RIT2 gene appeared in both the forward and reverse MR analyses (alcohol intake→PD: rs28597806, rs8083110; PD→alcohol intake: rs4588066). RIT2 is selectively expressed in the human brain (FPKM: 5.259±2.103), with low or no expression in peripheral tissues (FPKM: <1). Analysis of three human substantia nigra transcriptomic datasets revealed a decreasing trend in RIT2 gene expression in PD patients (GSE20141 array signal: 3.49±1.23 vs. 2.33±0.87, P=0.044). Animal experiments demonstrated that administration of 20% ethanol or 20% liquor (approximately 8% ethanol) stimulated a >2-fold upregulation of RIT2 gene expression in the mouse brain. Furthermore, transcriptomic sequencing revealed that the two alcohol-treated groups exhibited 96 (20% ethanol vs. water control) and 4 (20% liquor vs. water control) differentially expressed genes, respectively, indicating that low-dose alcohol consumption can achieve RIT2 upregulation while minimizing impact on other brain genes. In addition to its anti-infective effects, low-dose alcohol consumption primarily influences signaling pathways related to neurodegenerative diseases such as PD and Prion diseases.Conclusion Alcohol consumption is generally considered as a harmful lifestyle habit. However, some studies have also shown a lower risk of mortality among individuals who consume low doses of alcohol (100 g/week of ethanol) or drink occasionally. Currently, one of the research focuses on alcohol consumption is whether the human body can benefit from low-dose alcohol intake. This study provides new evidence supporting a negative association between alcohol consumption and PD, and for the first time, through MR analysis, identifies the RIT2 gene as a potential mediator of the effect of alcohol consumption on PD. RIT2 is selectively expressed in the human brain. Building upon existing evidence indicating downregulated RIT2 gene expression in PD pathogenesis, our experiments confirm that low-dose alcohol consumption can upregulate RIT2 expression in the brain. In brief, alcohol consumption may suppress the pathogenesis of PD by upregulating RIT2 expression in the substantia nigra. China is facing a serious problem of population aging. This study offers important insights for long-term PD prevention and treatment strategies, with the aim of benefiting more potential PD patients through lifestyle modifications, thereby improving the quality of life of the aging population and reducing the economic burden on healthcare.]]> 2026/5/23 0:00:00 LU Wei,CHENG Xiu-Li,PAN Xiao-Yun,YANG Dan-Dan,ZOU Hui-Ling,DONG Li-Guo,WEI Yi-Liang and CUI Gui-Yun 23true Objective This study aimed to elucidate the mechanistic role of Staphylococcus aureus in the pathogenesis of atopic dermatitis (AD), a chronic inflammatory skin disorder characterized by pruritus and barrier dysfunction. A key focus was screening traditional Chinese medicine (TCM) active components with dual antibacterial and antipruritic efficacy, followed by systematic evaluation of their in vitro antibacterial activity. Additionally, a novel drug delivery system was constructed to enable localized efficient drug delivery, inhibiting S. aureus proliferation and alleviating its induced pruritus, thereby providing new strategies for targeted AD therapy.Methods Male C57BL/6J mice aged 6-8 weeks (body weight 18-22 g) were used to establish an AD model via repeated oxazolone sensitization. On day 14, microbial samples were collected from the lesional area (1 cm2) using sterile cotton swabs, followed by vortex mixing, serial dilution, and plating on 5% sheep blood agar plates (incubated at 37°C for 24 h). Single colonies with complete transparent β-hemolytic zones were isolated and identified as vancomycin-intermediate S. aureus (VISA) via 16S rRNA sequencing. An S. aureus mono-infection animal model was then established by applying gauze saturated with bacterial suspension (McFarland turbidity 0.1) to the nape and back skin of mice. The pruritic phenotype and inflammatory cell infiltration induced by S. aureus were evaluated using comprehensive approaches including behavioral assays (e.g., scratching frequency recording), hematoxylin-eosin (HE) staining, and toluidine blue staining. The in vitro antibacterial efficacy of the TCM monomer pseudolaric acid B (PAB) and double network hydrogel (DN) was separately assessed by disk diffusion assay, while the minimum inhibitory concentration (MIC) of PAB was determined via broth dilution method. Further validation of the pharmacodynamic characteristics of the composite system (PAB@DN, composed of PAB and DN) was conducted through behavioral assays, HE staining, and dermatitis scoring, with its drug release profile evaluated by mass spectrometry analysis. Based on scratching behavioral analysis and dermatitis scoring, the optimal ratio and concentration of PAB@DN were optimized.Results The S. aureus load in AD lesional tissues was significantly higher than in normal skin ((5.3±0.33)×10? CFU vs. (3.6±0.26)×10? CFU, P<0.001). In the S. aureus mono-infection group, mice exhibited a 6.7-fold increase in scratching frequency compared to the control group. HE staining revealed marked epidermal thickening ((10.4±2.39) μm vs. (85.6±1.95) μm, P<0.000 1), and toluidine blue staining showed a 23-fold increase in mast cell degranulation. Pseudolaric acid B exhibited a significant concentration-dependent inhibitory effect on S. aureus growth, with its in vitro antibacterial effect being 57% that of the antibiotic cefepime (inhibition zone diameter: PAB (1.885±0.036) cm vs. cefepime (3.636±0.005) cm, P<0.000 1) and a minimum inhibitory concentration (MIC) of 1 g/L. The carrier double network hydrogel (DN) itself lacked direct antibacterial activity (no significant difference in inhibition zone diameter compared to the control) but effectively ameliorated the dry symptoms of AD-like lesions. The PAB@DN composite system demonstrated a synergistic effect compared to individual components, resulting in a 50% reduction in scratching behavior, an 86% decrease in dermatitis score, and a 60% reduction in epidermal thickening. It also reduced the S. aureus load in mouse skin by approximately 34%, with the optimal effective formulation being PAB at 1 g/L loaded onto DN.Conclusion S. aureus colonization plays a critical driving role in the onset and progression of AD. Using an S. aureus infection model, this study confirmed that the pseudolaric acid B-hydrogel composite delivery system (PAB@DN) can effectively alleviate S. aureus-induced pruritus and skin damage, providing experimental evidence for microbiota-targeted therapy of AD.]]> 2026/4/9 11:05:19 YOU Ye,YANG Yan,LI Tong-Yu,CAI Cheng-Long,WANG Ting,ZHU Chan and TANG Zong-Xiang 22true Objective The Golgi apparatus serves as a central hub in the eukaryotic secretory pathway, responsible for the processing, sorting, and trafficking of proteins and lipids. In mammalian cells, the Golgi typically forms a perinuclear ribbon-like structure composed of laterally connected cisternae stacks.The maintenance of Golgi ribbon structure depends on the balance of membrane flux across multiple intracellular trafficking pathways, yet the specific contributions of distinct trafficking branches to Golgi macroscopic morphology remain elusive. In mammalian cells, the Golgi ribbon is typically organized as a perinuclear, laterally connected structure composed of stacked cisternae, and its integrity is highly dynamic and sensitive to perturbations in membrane trafficking. This study aims to elucidate the role of coat protein complex I (COPI)-mediated retrograde transport in maintaining the Golgi ribbon and to dissect the functional relationship between the transmembrane cargo receptors LEPROT/LEPROTL1 (LEPROTs) and the COPI adaptor GOLPH3.Methods Using siRNA interference and gene-deficient cell lines, we selectively perturbed COPI- or adaptor protein complex 1 (AP-1)-mediated trafficking pathways in HeLa cells. To quantitatively evaluate Golgi morphology, we employed a “Golgi Angle”-based measurement to assess its circumferential distribution around the nucleus. The spatial distribution of the Golgi ribbon was quantitatively analyzed using confocal microscopy, while Golgi ultrastructure and vesicle density were examined via transmission electron microscopy. Additionally, the subcellular distribution of COPI components was assessed by immunofluorescence co-localization.Results Selective inhibition of COPI retrograde transport significantly induced the circumferential extension of the Golgi ribbon around the nucleus, whereas blocking AP-1-mediated anterograde transport resulted in Golgi compaction, indicating opposing roles. These results suggest that different trafficking branches downstream of ARF1 exert distinct and even antagonistic effects on Golgi morphology. LEPROTs-deficient cells exhibited a Golgi extension phenotype highly consistent with COPI impairment. Furthermore, knockdown of GOLPH3 in a LEPROTs double-knockout background produced a significant additive effect on Golgi extension, suggesting that LEPROTs and GOLPH3 play non-redundant roles in regulating COPI-related trafficking processes. Mechanistically, loss of either LEPROTs or GOLPH3 led to the aberrant accumulation of COPI components at endoplasmic reticulum exit sites, accompanied by a reduction in COPI-like vesicles around the Golgi. This redistribution indicates a defect in COPI recycling between the ER-Golgi interface and the Golgi apparatus. Ultrastructural analysis revealed that Golgi cisternae in defective cells became shorter and thicker while maintaining a stable number of stacks. In parallel, the density of Golgi-associated vesicles was markedly decreased, further supporting an impairment in COPI vesicle formation or budding processes.Conclusion This study demonstrates that active COPI retrograde transport is a critical factor in restricting the over-connection of the Golgi ribbon and maintaining its compactness. Rather than causing fragmentation, partial disruption of COPI function leads to a distinct morphological outcome characterized by Golgi ribbon extension at the light microscopy level and cisternal remodeling at the ultrastructural level. LEPROTs and GOLPH3 cooperatively promote the recycling and vesiculation of COPI components, thereby imposing a structural constraint on the Golgi periphery. Our findings support a model in which multiple adaptor proteins act in parallel to sustain efficient COPI cycling, thereby maintaining Golgi structural homeostasis. These findings provide new cell biological evidence for the membrane trafficking basis of Golgi morphological homeostasis.]]> 2026/4/8 11:42:02 GAO Jing-Hu,ZHAO Lin-Yue,ZHANG Yu-Lu,WU Yan-Fang and YAN Bing 21true Objective This study aims to construct a reconstituted high-density lipoprotein (rHDL) delivery system loaded with kinsenoside (KD@rHDL), and to systematically evaluate its function in enhancing the phagocytosis of amyloid β-protein (Aβ) by microglia and improving the inflammatory state of microglia, as well as to preliminarily explore its potential application value in the treatment of Alzheimer’s disease (AD).Methods KD@rHDL was prepared by the film hydration method combined with probe sonication and co-incubation. Its morphology was observed by transmission electron microscopy, and the particle size and Zeta potential were measured by dynamic light scattering. The encapsulation efficiency and drug loading were determined by high-performance liquid chromatography. The affinity between KD@rHDL and Aβ was analyzed by surface plasmon resonance (SPR) to assess its feasibility as a medium for Aβ clearance. At the cellular level, after treating mouse microglial cells (BV-2 cells) with KD@rHDL and adding fluorescently labeled Aβ, the phagocytic efficiency of microglia for Aβ was detected by confocal microscopy. Meanwhile, the CCK-8 method was used to evaluate the effect of KD@rHDL on cell viability to determine its safety. The trans-barrier transport ability of KD@rHDL was detected by Transwell assay. The expression levels of NLRP3 inflammasome and downstream inflammatory factor IL-1β in LPS-induced microglia were detected by Western blot to evaluate the regulatory effect of KD@rHDL on the inflammatory state of cells.Results Characterization results showed that the successfully prepared KD@rHDL presented a typical discoid structure under transmission electron microscopy, with a uniform particle size distribution, an average particle size of approximately (14.4±0.24) nm, and a suitable negative Zeta potential, demonstrating good colloidal stability. The drug content determination results indicated that the encapsulation efficiency of KD@rHDL for kinsenoside was (42.24±1.30)%, and the drug loading was (6.03±0.19)%, indicating a good drug loading capacity. The CCK-8 assay results showed that in the set concentration range, the survival rates of BV2 and HT22 cells in the KD@rHDL treatment group were all above 90%, with no significant difference from the control group, indicating good cell safety of the formulation. The results of the Aβ phagocytosis experiment indicated that, compared with the Aβ oligomers (Aβo) group alone, the fluorescence signal intensity within microglia in the KD@rHDL treatment group was significantly enhanced, and a large amount of fluorescence-labeled Aβ was observed to accumulate intracellularly under a fluorescence microscope. The SPR assay results showed that rHDL had a strong affinity for Aβ, with an affinity constant reaching the nanomolar level. Transwell assay results indicated that KD@rHDL could effectively cross the bEnd.3 cell monolayer barrier and be taken up by BV2 and HT22 cells. Western blot assay results showed that high-dose KD@rHDL treatment could significantly reduce the expression level of NLRP3 protein in LPS-induced microglia and simultaneously down-regulate the maturation and secretion of IL-1β, indicating that KD@rHDL can effectively inhibit the activation of the NLRP3 inflammasome pathway and improve the neuroinflammatory state mediated by microglia.Conclusion This study successfully constructed a reconstituted high-density lipoprotein delivery system loaded with kinsenoside (KD@rHDL). This nano-delivery system not only significantly enhances the phagocytic clearance ability of microglia towards Aβ, but also effectively inhibits the NLRP3/IL-1β-mediated inflammatory pathway, improving the inflammatory state of microglia. The above results indicate that KD@rHDL has a synergistic effect in promoting Aβ clearance and alleviating neuroinflammation, demonstrating potential therapeutic value for AD and providing new ideas and experimental basis for the development of subsequent AD treatment strategies.]]> 2026/5/21 8:57:04 CHEN Lu-Yao,MU Yan and HUA Qian 20true Objective The present study aimed to investigate alterations in white matter microstructure and spontaneous neural activity in male college smokers, and to further explore their associations with nicotine dependence. Given that adolescence and early adulthood represent critical periods for brain maturation, particularly for white matter development, understanding the neural correlates of smoking behavior during this stage is of substantial importance for both neuroscience and public health.Methods A total of 115 male undergraduate students were initially recruited for this study. After quality control and exclusion procedures, 52 male college smokers and 42 demographically matched healthy non-smokers were included in the final analysis. All participants underwent multimodal magnetic resonance imaging (MRI), including diffusion tensor imaging (DTI) and resting-state functional MRI (rs-fMRI). White matter fiber tracts were reconstructed using the automated fiber quantification (AFQ) method, which enables precise identification and quantification of major fiber bundles. Eighteen major white matter tracts were segmented for each participant. Along the core trajectory of each tract, 100 equidistant nodes were sampled. Fractional anisotropy (FA) was calculated at each node to assess white matter microstructural integrity, while amplitude of low-frequency fluctuation (ALFF) was computed to evaluate spontaneous neural activity within white matter tracts. Between-group differences in FA and ALFF were assessed using two-sample t-tests, with appropriate corrections applied for multiple comparisons. Furthermore, Pearson correlation analyses were conducted to examine the relationships between imaging-derived metrics (FA and ALFF values in regions showing significant group differences) and nicotine dependence severity, as measured by the Fagerstr?m test for nicotine dependence (FTND).Results Compared with healthy non-smokers, male college smokers exhibited significantly increased FA values in several white matter tracts, including the left thalamic radiation, right corticospinal tract, forceps major of the corpus callosum, left uncinate fasciculus, and right arcuate fasciculus. These findings suggest altered microstructural organization or increased directional coherence within these pathways. In addition, smokers demonstrated significantly elevated ALFF values in the forceps major, right uncinate fasciculus, and left arcuate fasciculus, indicating enhanced spontaneous neural activity in these white matter regions. Correlation analyses revealed that FA values in the left thalamic radiation and right corticospinal tract were negatively correlated with FTND scores, suggesting that higher levels of nicotine dependence were associated with reduced microstructural integrity or altered fiber organization in these regions. In contrast, ALFF values in the forceps major and right uncinate fasciculus were positively correlated with FTND scores, indicating that greater nicotine dependence was associated with increased spontaneous neural activity in specific white matter pathways.Conclusion The present study provides evidence that male college smokers exhibit distinct alterations in both white matter microstructure and functional activity. These abnormalities are not uniformly distributed but rather localized to specific fiber tracts implicated in sensorimotor processing, interhemispheric communication, and higher-order cognitive and emotional regulation. Importantly, the observed associations between imaging metrics and nicotine dependence severity suggest that these structural and functional alterations may reflect neurobiological mechanisms underlying addiction. The combination of AFQ-based tract profiling and multimodal MRI offers a sensitive approach for detecting subtle changes along white matter pathways, highlighting its potential utility in identifying neuroimaging biomarkers of nicotine dependence. Overall, these findings indicate that smoking during early adulthood may disrupt ongoing white matter maturation, potentially leading to long-term consequences for brain function. This study provides novel insights into the neural basis of nicotine dependence and underscores the importance of early intervention and prevention strategies targeting young smokers.]]> 2026/4/9 10:20:48 LI Xiao-Jiao,YU Da-Hua,XUE Ting,YUAN Kai,MAI Zhen-Zhen,WANG Xu-Wen,DONG Fang,WANG Juan and MA Yu-Xin 19true E. coli). To compare the differential magnetic responses among bacteria expressing various recombinant MagR proteins, we utilized high-throughput automated bright-field microscopic imaging and scanning electron microscopy (SEM). Furthermore, comprehensive biochemical and biophysical characterizations of iron and iron-sulfur cluster binding were performed using Ferrozine colorimetric assays, electron paramagnetic resonance (EPR) spectroscopy, ultraviolet-visible (UV-Vis) absorption, and circular dichroism (CD) spectroscopy. Additionally, 100 mT static magnetic field (SMF) exposure experiments were conducted to assess magnetically tunable phenotypes, while the intrinsic magnetic properties of purified MagR proteins were directly measured using a superconducting quantum interference device (SQUID) magnetometer. Results Our results demonstrated that the heterologous expression of MagR homologs induced varying degrees of bacterial filamentation. From this comprehensive screen, two distinct morphological patterns were identified: hydra (Hydra vulgaris) MagR (hyMagR) promoted uniform cell elongation and filamentation, exhibiting robust magnetic sensitivity manifested as significantly enhanced filamentation under the 100 mT SMF. In contrast, pigeon (Columba livia) MagR (clMagR) induced only low-frequency, extreme filamentation (sporadically exceeding 80 μm) with a relatively weaker magnetic morphological response. Mechanistically, our data unambiguously proved that these phenotypic differences are primarily driven by distinct iron redox preferences rather than total cellular iron accumulation. Specifically, hyMagR preferentially binds ferrous iron (Fe2+), whereas clMagR favors ferric iron (Fe3+) and forms more stable iron-sulfur clusters. Intriguingly, although SQUID magnetometry showed that purified clMagR exhibited approximately five-fold higher mass magnetic susceptibility than hyMagR, its cellular magnetic response was weaker. We hypothesize that the Fe2+-preferred intracellular environment associated with hyMagR overexpression primes the cell for enhanced generation of reactive oxygen species (ROS) via the Fenton reaction. Exposure to an SMF synergizes with this primed redox state, triggering the bacterial SOS response and upregulating cell division inhibitors to efficiently induce uniform filamentation. Conclusion Our findings identify the Fe2+/Fe3+ redox state as a critical determinant of MagR-mediated morphological remodeling and magnetic responsiveness. This discovery suggests a potential strategy for engineering magnetically responsive cellular systems for synthetic biology applications, and provides a plausible framework, which potentially combines intrinsic protein magnetism with redox-state modulation, for further investigating the evolutionary mechanisms of MagR-mediated magnetoreception.]]> 2026/3/26 0:00:00 CHANG Ya-Fei,ZHANG Jing,ZHANG Peng,ZHOU Xiu-Juan,WEI Meng-Ke,CAI Tian-Tian,HE Pei-Qi,WANG Jun-Feng and XIE Can 18true Objective Transcranial magneto-acoustic stimulation (TMAS) is an emerging non-invasive neuromodulation technique that may provide a novel non-pharmacological intervention strategy for Parkinson""s disease (PD). PD is characterized by the progressive degeneration of dopaminergic neurons in the substantia nigra pars compacta (SNc), leading to motor impairments such as bradykinesia, tremor, and rigidity. Increasing evidence indicates that mitochondrial dysfunction and impaired mitochondrial quality control are central mechanisms underlying dopaminergic neuronal loss. In particular, abnormalities in mitophagy and mitochondrial fission-fusion balance contribute substantially to oxidative stress, energy metabolic failure, and neuronal injury. At present, most clinical treatments for PD mainly alleviate symptoms but do not effectively halt disease progression. Therefore, exploring new interventions targeting the core pathological mechanisms is of considerable significance. This study aims to investigate whether TMAS can improve neural damage and motor dysfunction in PD mice by regulating mitophagy and the fission/fusion dynamic balance, thereby providing theoretical and experimental support for its application in PD treatment.Methods Male C57BL/6 mice were used in this study. A PD model was established by intraperitoneal injection of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) for 7 consecutive days. After model induction, mice in the intervention group received TMAS once daily for 14 consecutive days, whereas the corresponding control group received sham stimulation. The stimulation target was positioned over the primary motor cortex (M1). Motor performance was evaluated using the pole test and the open-field test. To verify the activation effect of TMAS on the target cortical region, c-Fos immunohistochemistry was performed in the M1. To assess nigral dopaminergic neuronal injury, tyrosine hydroxylase (TH) immunohistochemistry was used to quantify TH-positive neurons in the SNc. Mitochondrial function was evaluated by measuring reactive oxygen species (ROS) levels and adenosine triphosphate (ATP) content in the SNc. Western blot was further performed to determine the expression of mitophagy-related proteins, including PINK1, Parkin, LC3-II, and p62, as well as mitochondrial dynamics-related proteins, including Drp1 and Opa1.Results TMAS significantly increased the number of c-Fos-positive cells in M1 (P<0.000 1), indicating effective activation of neurons in the targeted cortical region. Compared with the control group, MPTP-treated mice exhibited marked motor dysfunction, including a significant reduction in total distance traveled in the open-field test (P<0.000 1) and mean speed (P=0.000 1), as well as significant prolongation of turn time and total climbing time in the pole test (P<0.000 1). These behavioral impairments were accompanied by a substantial loss of TH-positive dopaminergic neurons in the SNc, whereas TMAS significantly increased TH-positive neuron survival (P<0.000 1). In parallel, MPTP induced a pronounced increase in ROS levels and a significant reduction in ATP content, indicating severe mitochondrial dysfunction and energy metabolism impairment (P<0.01). TMAS treatment significantly improved motor performance, as reflected by the reversal of MPTP-induced impairment in the open-field and pole tests, and significantly reduced ROS accumulation (P<0.01) while restoring ATP production (P<0.001). At the molecular level, MPTP markedly downregulated PINK1 and Parkin, decreased p62 expression, increased LC3-II accumulation, elevated Drp1 expression, and reduced Opa1 expression, whereas TMAS significantly reversed these abnormalities, suggesting restoration of mitophagy-related mitochondrial quality control and re-establishment of mitochondrial fission-fusion balance. Collectively, these findings indicate that TMAS ameliorates MPTP-induced neurotoxicity and restores mitochondrial homeostasis and energy metabolism.Conclusion TMAS effectively attenuates neural damage and improves motor dysfunction in MPTP-induced PD mice. Its neuroprotective effects are closely associated with multidimensional regulation of the mitochondrial quality control system, including restoration of PINK1/Parkin-mediated mitophagy and rebalancing of Drp1/Opa1-related mitochondrial dynamics. Rather than acting only as a symptomatic neuromodulatory intervention, TMAS may influence a key pathological axis of PD by improving mitochondrial homeostasis in SNc and protecting nigral dopaminergic neurons. These findings provide experimental evidence supporting TMAS as a promising non-invasive physical intervention for PD.]]> 2026/4/3 11:25:24 ZHANG Shuai,WANG Yan-Bin,XU Yi-Hao,MI Jin-Rui,LU Xiao-Chao,AN Yu-Chen,LIU Ji-Zhou and SUN Jia-Qi 17true Objective Mitochondria are not only the central organelles responsible for cellular energy metabolism but also play essential roles in regulating cell cycle progression and cytoskeletal dynamics. In recent years, accumulating evidence has demonstrated that mitochondrial homeostasis is closely associated with mitotic progression and cytokinesis. Schizosaccharomyces pombe serves as a classical and well-established model organism. Because its cell cycle regulatory mechanisms are highly conserved throughout evolution, its genetic background is clearly defined, and experimental manipulation is efficient and convenient, it has been extensively applied in studies of cell growth, division, and reproductive mechanisms. The SPBC1604.04 gene encodes a previously uncharacterized mitochondrial carrier protein in Schizosaccharomyces pombe. This gene is located on chromosome II and spans 1 018 base pairs in length. It encodes a protein consisting of 238 amino acids with a predicted molecular mass of approximately 31.03 ku. Bioinformatic analysis predicts that this protein is responsible for the transport of thiamine pyrophosphate (TPP) into mitochondria. However, the effects of SPBC1604.04 gene deletion on mitotic cell dynamics under different temperature conditions have not been fully elucidated.Methods The SPBC1604.04 deletion strain of Schizosaccharomyces pombe was used as the experimental model. Fluorescent protein markers were constructed in the deletion background to label mitochondria, microtubules, actin, myosin, the nuclear envelope, and chromosomes. Live-cell imaging was performed using a TCS-SP8 laser scanning confocal microscope under normal temperature conditions (25℃) and heat stress conditions (37℃). Time-lapse microscopy was applied to dynamically monitor mitochondrial morphology and distribution, spindle assembly and elongation, chromosome segregation, as well as the formation and constriction of the actomyosin ring during cytokinesis. ImageJ software was used for quantitative measurements, including microtubule length during mitosis, spindle length at different mitotic stages, mitochondrial fluorescence intensity as an indicator of mitochondrial content, actomyosin ring length, nuclear envelope area, and chromosome segregation timing. Statistical analyses were conducted to compare phenotypic differences between the wild-type and SPBC1604.04 deletion strains at both temperature conditions. Through these analyses, we systematically investigated the impact of SPBC1604.04 deletion on mitotic cell dynamics in fission yeast under both normal physiological conditions and temperature stress.Results At 25℃, compared with wild-type cells, the SPBC1604.04Δ strain exhibited a pronounced tendency toward mitochondrial fragmentation, accompanied by abnormal mitochondrial content and a significant reduction in mitochondrial fluorescence intensity. These observations suggest impaired mitochondrial homeostasis under normal growth conditions. In addition, the constriction time of actomyosin ring during cytokinesis was markedly prolonged, indicating that deletion of SPBC1604.04 affects the dynamics of the contractile machinery. However, no obvious defects were observed in spindle assembly, spindle elongation, or chromosome segregation. Under heat stress at 37℃, mitochondrial morphology in the SPBC1604.04Δ strain showed a tendency to recover toward a continuous tubular network structure. Mitochondrial content was restored, fluorescence intensity increased, and the constriction time of the actomyosin ring returned to levels comparable to those of wild-type cells. These results indicate that the mitotic defects observed at normal temperature are partially or fully alleviated under heat stress conditions.Conclusion This study demonstrates that deletion of the SPBC1604.04 gene leads to abnormal mitochondrial content in Schizosaccharomyces pombe. The mitochondrial carrier protein SPBC1604.04 participates in regulating actomyosin ring constriction during mitosis but does not appear to be directly involved in the regulation of spindle dynamics or chromosome segregation. Our findings provide key experimental evidence for understanding the functional link between the SPBC1604.04 gene, mitochondrial homeostasis, and mitotic regulation.]]> 2026/3/24 0:00:00 XU Jia-Ni,HE Jia-Yi,ZHENG Lang-Lin,HE Shu-Rong,MA Shuai,DING Xiang and HOU Yi-Ling 16true Objective The malaria parasites remodel the host erythrocyte structure by exporting parasite proteins that interact with the membrane skeleton proteins of red blood cells (RBCs), facilitating their intracellular survival and pathogenicity. Skeleton-binding protein 1 (SBP1) is a conserved exported protein across Plasmodium species. In Plasmodium falciparum, SBP1 has been reported to interact with erythrocyte membrane skeleton proteins 4.1R and spectrin, while its contribution to erythrocyte remodeling and parasite virulence in Plasmodium berghei (Pb) remains unclear. This study aims to determine whether PbSBP1 associates with the host cytoskeletal protein 4.1R and to investigate its role in the remodeling of host RBCs and the pathogenicity of Plasmodium berghei.Methods In Plasmodium berghei, the relationship between PbSBP1 and the erythrocyte cytoskeletal protein 4.1R was examined using co-immunoprecipitation. A Pbsbp1 gene knockout mutant of Plasmodium berghei (Pbsbp1?) was generated based on the principle of double crossover homologous recombination. The deformability of erythrocytes infected with Pbsbp1? parasites was assessed using microfluidic methods. Microchannels with an array of cylindrical pillars were used to detect modifications in infected RBC deformability. The infected RBCs were squashed between the rows and recovered between the columns and the transit velocity (μm/s) of infected RBCs travelling through the microchannel was recorded. The component of the erythrocyte membrane skeleton junctional complex, tropomodulin (TMOD), was fluorescently labeled, and the cytoskeletal network of infected erythrocytes was imaged using super-resolution stochastic optical reconstruction microscopy (STORM) to analyze ultrastructural changes in the cytoskeleton of wild-type (WT) and Pbsbp1?-infected erythrocytes. Actin-based junctional complexes were displayed as individual clusters by the labeled TMOD in the STORM images, and the cluster densities and distances between adjacent clusters of infected RBCs were calculated. Additionally, rodent malaria models (BALB/c mice) and experimental cerebral malaria models (C57BL/6 mice) were employed to monitor the growth of Pbsbp1? and WT parasites during the intraerythrocytic stage and their capacity to induce cerebral malaria in mice.Results PbSBP1 may participate in the remodeling of infected erythrocytes through direct or indirect interaction with the erythrocyte cytoskeletal protein 4.1R. Microfluidic assays revealed that the deformability of erythrocytes infected with Pbsbp1? parasites was significantly enhanced compared to those infected with WT parasites. STORM imaging further demonstrated that the ultrastructure of the erythrocyte cytoskeleton in Pbsbp1?-infected cells was altered relative to that in WT-infected erythrocytes. The distances between nearest neighbors of clusters had a tendency to increase while the cluster densities were decreased in Pbsbp1?-infected RBCs compared to WT-infected RBCs. Subsequent phenotypic analysis indicated that the growth rate of Pbsbp1? parasites during the intraerythrocytic stage was significantly slower than that of WT parasites, and their ability to induce cerebral malaria in mice was also attenuated. These findings suggest that PbSBP1 is involved in the remodeling of the erythrocyte membrane skeleton, likely through its direct or indirect interaction with protein 4.1R, thereby regulating the deformability of infected erythrocytes and influencing the pathogenicity of the blood-stage parasites.Conclusion This study establishes a role for PbSBP1 in host erythrocyte remodeling and parasite virulence, providing new research strategies for the prevention and treatment of malaria.]]> 2026/3/1 21:36:43 GUO Xin-Yue,ZHAO Huan-Qi,ZHONG Yan-Xuan,JIANG Ru-Meng,LI Yao-Xian,PAN Lei-Ting,WANG Qian and SHI Xiao-Yu 15true Objective Zheng’s San Qi San (ZSQS) power, a classic traditional Chinese medicine (TCM) formula, is used for treating soft tissue injuries involving muscles, tendons, and ligaments. However, its underlying therapeutic mechanisms remain unclear. This study aimed to screen and identify pharmaceutically active ingredients and their candidate biomolecule targets, and further elucidate the molecular mechanism of ZSQS in the treatment of skeletal muscle injury.Methods Network pharmacology was employed to construct “ZSQS-component-target”, “protein-protein interaction (PPI)” and “active ingredient-core protein-pathway” networks to predict the key active ingredients and potential core targets of ZSQS for skeletal muscle injury. The predicted results were then validated via microarray data from the GEO database. Molecular docking was then performed to assess the binding ability between the screened active ingredients of ZSQS and the candidate core targets. Moreover, liquid chromatography-mass spectrometry (LC-MS) was used for qualitative and quantitative analysis to verify the active components of the drug and ZSQS serum. Finally, an animal model of eccentric exercise-induced skeletal muscle injury and a myotube cell model of oxidative stress-induced injury were established to validate the effects of ZSQS and its interventional effects on the biological functions of critical targets, thereby demonstrating the potential therapeutic mechanism of ZSQS.Results Among the 111 active components identified in ZSQS and their corresponding 204 targets related to the skeletal muscle injury repair process, 14 core targets (including AKT1) and 4 core active components (quercetin, luteolin, kaempferol, and β-sitosterol) were screened out, while the corresponding metabolites of quercetin, luteolin and kaempferol were detected in the ZSQS serum. Among these targets, 5 candidate genes (IL-6, CASP3, HIF1A, STAT3, and JUN) overlapped with the differential expression screening results with GEO data, and IL-6 was confirmed to be enriched in the PI3K/AKT pathway. Combined with the prediction results of the AKT expression levels, these findings suggest that the phosphorylation level of AKT1 plays a core role in the therapeutic mechanism of ZSQS. Molecular docking analysis further revealed that the PH domain of AKT1 had high binding energy with all 4 core active components, as verified by LC-MS. Finally, animal model studies have shown the promoting effect of ZSQS administration on skeletal muscle injury repair and its possible antioxidant damage mechanism. Cell model studies further demonstrated that ZSQS-containing serum, core active ingredient combination therapy, and quercetin monomer could increase the phosphorylation level of AKT, promote the nuclear translocation of Nrf2, upregulate the expression of downstream antioxidant enzymes (SOD, GPx, and GR), and inhibit the expression of inflammatory factors (IL-6 and TNF-α), thereby alleviating oxidative stress and the inflammatory response.Conclusion ZSQS alleviates skeletal muscle injury mainly by activating the AKT/Nrf2 signaling pathway, enhancing cellular antioxidant and anti-inflammatory capabilities. The results of this study provide a scientific basis for the clinical application and modernized development of ZSQS.]]> 2026/1/23 15:27:52 WANG Ding-Rui,LIU Yun-Xin,XU Jun-Jie,YANG Liu,Lü Jia-Hao,XING Cheng-Yuan,Lü Lei and QIE Bei-Bei 14true Objective The exacerbating trend of global population aging poses profound socioeconomic and public health challenges, making the comprehensive elucidation of biological aging mechanisms and the discovery of effective anti-aging interventions an urgent priority in the life sciences. Based on our previous serum metabolomics findings that dimethylglycine, an intermediate metabolite of amino acid metabolism naturally present in the human body, was significantly enriched in the serum of longevity families, this study aimed to systematically investigate the anti-aging effects of dimethylglycine both in living organisms and in controlled laboratory environments, and to preliminarily elucidate its underlying molecular mechanisms. While existing literature indicates that dimethylglycine possesses antioxidant and immunomodulatory properties, its direct anti-aging efficacy and the specific molecular pathways through which it operates remain largely unexplored.Methods To comprehensively evaluate the anti-aging properties of dimethylglycine, we utilized replicative senescent human embryonic lung fibroblasts, specifically the WI-38 cell line, as an experimental model in a controlled laboratory environment. Cell viability and safety were thoroughly assessed using Cell Counting Kit-8 and lactate dehydrogenase release assays across various concentrations of dimethylglycine. The impact of dimethylglycine on cellular senescence phenotypes, oxidative stress, and proliferative capacity was evaluated via senescence-associated beta-galactosidase staining, reactive oxygen species fluorescence detection, and 5-ethynyl-2"-deoxyuridine incorporation assays. Furthermore, the molecular alterations of senescence-associated secretory phenotype factors and core senescence signaling pathways were quantified using quantitative reverse transcription polymerase chain reaction for the messenger RNA levels of interleukin-6, interleukin-8, p21, and matrix metalloproteinase-1, and enzyme-linked immunosorbent assay for the measurement of p16 and p21 protein expression levels. For the living organism model, the wild-type nematode Caenorhabditis elegans was used to evaluate systemic physiological effects. We conducted a comprehensive lifespan analysis at 20°C, heat stress resistance survival assays at 35℃, senescence-associated beta-galactosidase staining, lipofuscin accumulation tracking, intracellular reactive oxygen species measurement, and Oil Red O staining to ascertain systemic lipid accumulation. Additionally, network pharmacology bioinformatics tools, including PharmMapper and STRING databases, and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis were utilized to predict target pathways, alongside highly detailed molecular docking simulations utilizing SwissDock and Protein-Ligand Interaction Profiler to examine interactions with the cytochrome P450 family 2 subfamily C member 9 protein.Results The experimental outcomes robustly demonstrate the potent anti-aging capabilities of dimethylglycine. At the cellular level, toxicity analyses firmly confirmed that dimethylglycine is highly safe; continuous treatment with 50 mol/L and 70 mol/L of dimethylglycine for 5 d did not induce any cellular membrane damage or cytotoxicity, but rather actively promoted cellular proliferation. Utilizing the optimal standardized concentration of 50 mol/L, dimethylglycine treatment significantly ameliorated senescent phenotypic markers in human embryonic lung fibroblasts, which was evidenced by a drastic and highly significant reduction in the senescence-associated beta-galactosidase positive cell percentage (P<0.000 1) and intracellular reactive oxygen species levels (P<0.000 1), alongside a marked increase in the 5-ethynyl-2"-deoxyuridine-positive proliferation rate (P=0.003 5). On a molecular expression scale, dimethylglycine significantly downregulated the messenger RNA expression of multiple core senescence-associated secretory phenotype inflammatory factors, including interleukin-6, interleukin-8, p21, and matrix metalloproteinase-1. Concurrently, it effectively suppressed the protein expression of critical cell cycle arrest markers, diminishing p16 protein levels by 57.3% (P=0.000 4) and p21 protein levels by 27.2% (P=0.000 7). In the nematode Caenorhabditis elegans animal model, dimethylglycine significantly extended the mean lifespan from 20.402 d to an impressive 23.066 d (P<0.000 1) and notably enhanced overall survival rates under severe heat stress environmental conditions (P=0.017). Furthermore, systemic dimethylglycine intervention significantly mitigated age-related physiological decline by decreasing bodily lipofuscin accumulation (P<0.000 1), significantly reducing senescence-associated beta-galactosidase activity, lowering systemic reactive oxygen species fluorescence (P=0.008), and effectively alleviating overall fat accumulation (P<0.000 1). Mechanistically, extensive network pharmacology and Kyoto Encyclopedia of Genes and Genomes analyses strongly revealed that the potential targets of dimethylglycine are significantly enriched in fundamental drug metabolism and oxidative stress response pathways. Precision molecular docking simulations conclusively demonstrated that dimethylglycine forms highly stable structural interactions with the cytochrome P450 family 2 subfamily C member 9 protein, specifically highlighting the definitive formation of 5 stable hydrogen bonds involving serine 365, leucine 366, and serine 429 residues, as well as two critical salt bridge formations with arginine 97 and histidine 368 residues. It is additionally predicted to interact favorably with glutathione S-transferase family proteins.Conclusion Dimethylglycine exhibits a profoundly significant and multifaceted anti-aging activity at both the cellular and entire living animal levels. By powerfully alleviating oxidative stress, heavily suppressing the core p16 and p21-dependent cellular senescence signaling pathways, and substantially mitigating the detrimental senescence-associated secretory phenotype, dimethylglycine effectively delays fundamental cellular senescence processes and drastically extends whole-organism lifespan. The biological mechanisms driving these robust protective effects are highly likely closely associated with its direct stable interactions with crucial metabolic and detoxifying enzyme systems, such as cytochrome P450 family 2 subfamily C member 9 and glutathione S-transferase family proteins, thereby systemically improving metabolic dysregulation and restoring critical redox homeostasis. This comprehensive study provides highly solid experimental evidence supporting dimethylglycine as a highly potent and safe potential anti-aging intervention agent, while simultaneously offering a clear molecular mechanistic explanation for the previously documented high abundance of dimethylglycine observed within exceptionally long-lived human populations.]]> 2026/3/17 15:43:32 HU Jie,PU Gong-Yu,LI Jun-Lin,CAO Ju,LIN Zhi-Xin,AN Wei-Wei,LI Xue-Meng and AN Jing 13true Objective Gastric hemorrhage is one of the most common and life-threatening emergencies of the upper digestive tract. Early identification and continuous monitoring are essential for reducing rebleeding rates and mortality, particularly within the critical early hours after onset. Although endoscopy and radiological imaging can accurately localize bleeding sites, these approaches are invasive, resource-intensive, and unsuitable for continuous bedside monitoring. Electrical impedance tomography (EIT), as a noninvasive and radiation-free functional imaging technique, offers real-time visualization of conductivity distribution and has the potential for detecting intragastric bleeding based on the electrical contrast between blood and surrounding gastric tissues. In this study, a three-dimensional gastric EIT (3D-gEIT) framework is proposed to achieve noninvasive, real-time, and dynamic monitoring of gastric hemorrhage, with emphasis on spatial localization and quantitative volume assessment.Methods A three-dimensional upper-abdominal simulation model incorporating the stomach, gastric wall, gastric contents, and surrounding tissues was established. Three electrode configurations, namely the dual layer ring, the four layer staggered ring, and the opposed dual plane array, were designed and systematically compared to evaluate their influence on depth sensitivity and spatial resolution. Based on the Tikhonov-Noser hybrid regularization scheme, a region-clustering constraint was introduced to develop the TK-Noser-RCC algorithm. This approach aggregates spatially adjacent elements with similar conductivity variations, thereby enhancing structural continuity and suppressing isolated noise artifacts. To validate the proposed framework, an upper-abdominal physical phantom was constructed using agar to simulate background tissue conductivity. Hemispherical high-conductivity inclusions with volumes ranging from 10 ml to 50 ml were attached to the inner gastric wall to mimic localized bleeding under different gastric filling states. Boundary voltages were acquired under a 120 kHz excitation current and reconstructed using the TK-Noser-RCC algorithm. Furthermore, an in vivo animal experiment was performed using a porcine model with adult-scale abdominal dimensions. A total of 100 ml of autologous blood was injected incrementally into the stomach to simulate progressive gastric hemorrhage, and time-difference EIT reconstruction was conducted at each injection stage to assess the dynamic system response under physiological conditions.Results Simulation results demonstrated that the opposed dual-plane electrode array achieved superior depth sensitivity distribution and spatial resolution. For a 40 ml hemorrhage model, the average ICC and SSIM improved by 55.9% and 38.8% compared with the dual-layer ring configuration, and by 64.0% and 39.5% compared with the four-layer staggered configuration. The proposed region-clustering constraint significantly enhanced reconstruction stability. Under added Gaussian noise of 40 dB and 30 dB, ICC values remained approximately 0.85, indicating effective artifact suppression and preservation of boundary integrity. In physical phantom experiments, reconstructed hemorrhage volumes increased approximately linearly with the preset hemispherical volumes, and the reconstructed high-conductivity regions closely matched the actual bleeding locations. Both empty-stomach and full-stomach conditions were evaluated, demonstrating that the opposed dual-plane configuration maintained stable imaging performance across varying gastric contents. In the animal experiment, reconstructed low-impedance regions expanded progressively with increasing injected blood volume. The spatial localization of the hemorrhage remained stable throughout the procedure, and no significant artifacts were observed. Quantitative analysis showed that reconstructed volume and average conductivity variation exhibited an approximately linear growth trend with injected blood volume, confirming the sensitivity of the system to dynamic intragastric conductivity changes.Conclusion The proposed 3D-gEIT framework enables quantitative reconstruction of gastric hemorrhage volume and spatial distribution with improved depth sensitivity, structural continuity, and noise robustness compared with conventional EIT approaches. By integrating optimized electrode configuration and a region-clustering-constrained reconstruction algorithm, the system provides stable dynamic monitoring under both controlled phantom conditions and in vivo physiological environments. This method offers a noninvasive, real-time, and low-cost imaging strategy for early diagnosis, postoperative monitoring, and bedside surveillance of gastric bleeding.]]> 2026/2/26 9:22:55 ZHAO Zi-Han,SUN Bo,HUANG Jing-Shi,LI Zhi-Wei,WU Yang,LI Nan,YAO Jia-Feng and ZHAO Tong 12true Objective Protein-protein interactions (PPIs) are fundamental to the execution of biological functions within living cells. However, traditional biochemical methods, such as co-immunoprecipitation (Co-IP), often fail to capture transient, weak, or membrane-associated interactions due to the stringent detergent requirements for cell lysis. Proximity labeling (PL) has emerged in recent years as a transformative technology for mapping the proteomes of specific subcellular compartments and identifying dynamic interactomes in situ. Golgi protein 73 (GP73, also known as GOLPH2), a resident type II Golgi transmembrane protein, is a well-recognized clinical biomarker for liver diseases, including hepatocellular carcinoma (HCC). Despite its clinical significance, the comprehensive physiological and pathological functions of GP73 remain partially understood. This study aims to establish an APEX2-mediated proximity labeling system specifically targeting GP73 to map its interactome in a living cellular environment, thereby providing new insights into its molecular roles and regulatory mechanisms.Methods To achieve spatial specificity, we first constructed a stable cell line expressing a fusion protein consisting of GP73 and the engineered soybean peroxidase APEX2. The localization of the GP73-APEX2 fusion protein was validated to ensure it correctly targeted the Golgi apparatus. The proximity labeling reaction was initiated by incubating the cells with biotin-phenol (BP) for 30 min, followed by a brief (1 min) treatment with 1 mmol/L hydrogen peroxide (H2O2). This catalytic reaction converts BP into highly reactive, short-lived biotin-phenoxyl radicals that covalently attach to endogenous proteins within a small labeling radius of the GP73-APEX2 enzyme. Subsequently, the cells were quenched, and biotinylated proteins were enriched using high-affinity streptavidin-coated magnetic beads. The captured “neighbor” proteins were subjected to on-bead digestion and analyzed via liquid chromatography-tandem mass spectrometry (LC-MS/MS) for high-throughput identification. Rigorous bioinformatics analysis, including Gene Ontology (GO) enrichment, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis, and protein-protein interaction network mapping, was performed to interpret the biological significance of the identified candidates.Results Our results demonstrate the successful establishment of a robust and sensitive APEX2-based proximity labeling system for GP73. We identified a total of 95 high-confidence interacting proteins that were significantly enriched in the GP73 proximity proteome compared to control groups. Bioinformatics analysis revealed that these interactors were predominantly associated with biological processes such as vesicular transport, protein localization, and, most notably, molecular functions related to “ribosome binding” and “translation regulation”. This suggested an unexpected role for the Golgi-resident GP73 in the cellular translation machinery. To validate these findings, we performed targeted biochemical assays which confirmed a direct interaction between GP73 and the subunits of the eukaryotic translation initiation factor 3 (eIF3) complex, specifically EIF3G and EIF3I. Furthermore, functional validation using the surface sensing of translation (SUnSET) assay—a non-radioactive method to monitor protein synthesis—revealed that the overexpression of GP73 significantly promoted global protein translation levels in the cell, whereas its depletion or inhibition resulted in reduced translation efficiency.Conclusion This study successfully utilized APEX2-mediated proximity labeling to provide the first systematic map of GP73 interactome in living cells. Our findings uncover a novel, unconventional function of GP73 as a regulator of cellular protein translation, likely mediated through its interaction with the eIF3 complex. This discovery significantly broadens our understanding of the biological roles of GP73 beyond its traditional function in the Golgi apparatus and suggests that it may act as a bridge between Golgi-related trafficking and the protein synthesis machinery. Furthermore, the technical framework established in this study provides a valuable template for investigating other complex organelle-associated protein networks and resolving transient macromolecular interactions in various physiological and pathological contexts.]]> 2026/2/4 9:25:54 LIU Mu-Yi,ZHANG Chang,YANG Meng-Xin,YAN Xin-Long,WAN Lu-Ming and WEI Cong-Wen 11true Objective To explore the effect of oral sodium butyrate on skeletal muscle atrophy in CT26 tumor mice through the gut microbiota-skeletal muscle axis and its potential mechanism.Methods Sixty SPF BALB/c male mice aged 8 weeks were randomly divided into a normal control group (NC, n=18) and a ABX-depleted group (ABX, n=42). The ABX mice were pretreated with a quadruple antibiotic cocktail via oral gavage (0.2 ml per administration, once daily, 6 d per week, for 2 weeks), whereas NC received an equal volume of sterile water. The quadruple antibiotic cocktail consisted of metronidazole (1 g/L), vancomycin (0.5 g/L), ampicillin (1 g/L), and gentamicin (1 g/L). Following successful pretreatment, six mice from each group were randomly selected for gut microbiota sequencing analysis and designated as the Abx group and the NC0 group, respectively. Theremaining mice in ABX were subcutaneously inoculated in the dorsum with 0.2 ml of CT26 cell suspension (at a cell density of 1×107/ml). Then these mice were randomly allocated into three subgroups: a control tumor bearing model group (0_NaB, n=12), a tumor-bearing model group receiving low-dose oral sodium butyrate (L_NaB, n=12), a tumor-bearing model group receiving high-dose oral sodium butyrate (H_NaB, n=12). And mice in NC were inoculated at the same site with 0.2 ml of normal saline. The administration dose for L_NaB was Remaining 0.3 g/(kg·d), that for H_NaB was 0.5 g/(kg·d), while NC and 0_NaB were given the same volume of normal saline (0.2ml per time, once daily, 6 d per week, for 4 weeks). The general condition of mice was monitored, and forelimb grip strength gastrocnemius muscle mass and its muscle fiber cross-sectional area were measured for each group. The structural changes in gut microbiota were assessed by 16S rRNA sequencing of cecal contents. Pathological alterations in the intestinal wall were examined via HE staining. Serum and gastrocnemius muscle levels of TNF-α, IL-6, IL-1β, and LPS were quantified using ELISA. The protein expression of ZO-1 and occludin in the small intestine, as well as proteins associated with the TLR4/MyD88/NF-κB signaling pathway in the gastrocnemius muscle, were detected by Western blot analysis.Results (1) The alpha-diversity in Abx was significantly lower than that in NC0 (P<0.01), a significant decrease of the mass and muscle fiber cross-sectional area of the gastrocnemius (P<0.01), with the majority of gut microbiota being effectively depleted. (2) Compared with NC, the subcutaneous tumors of mice in 0_NaB were prominent, a significant increase of the mass and muscle fiber cross-sectional area of the gastrocnemius, accompanied by a significant decrease in body weight at the end of the 3th and 4th week (P<0.05), and a significant weakening of the forelimb grasping strength at the 5th and 6th week (P<0.01). Compared with 0_NaB, the tumor mass of mice in L_NaB and H_NaB showed a significant decreasing trend, and the grip strength of the forelimbs significantly increased at the 5th and 6th week (P<0.05, P<0.01). (3) Compared with 0_NaB, the Shannon and Observed species indices in α diversity of L_NaB and H_NaB were significantly increased (P<0.05). At the genus level, compared with 0_NaB, L_NaB exhibited a significant decrease in the relative abundance of Parasutterella (P< 0.01), while H_NaB showed significant reductions in the relative abundances of both Escherichia-Shigella and Parasutterella (P < 0.01). (4) Compared with 0_NaB, the small intestinal tissue structure in L_NaB and H_NaB was more intact, the infiltration of inflammatory cells was significantly reduced, and the capillaries were slightly dilated. The expression levels of ZO-1 and occludin proteins in L_NaB were significantly increased (P<0.01). (5) The LPS concentration in the gastrocnemius muscle and the protein expression levels of TLR4, MyD88, p-IκBα, and p-NF-κB p65 in L_NaB and H_NaB were significantly lower than those in 0_NaB (P<0.05). The serum TNF-α concentration in H_NaB and TNF-α concentration in the gastrocnemius muscle of the L_NaB and H_NaB were significantly lower than those in 0_NaB (P<0.05, P<0.01, P<0.01).Conclusion Oral administration of NaB can improve gut microbiota α diversity, adjusting its composition, improving intestinal mucosal barrier function, reducing the LPS-induced pro-inflammatory response, and delaying skeletal muscle atrophy. The underlying mechanism may involve down regulation of TLR4/MyD88/NF-κB signaling in skeletal muscle.]]> 2026/2/6 16:34:46 ZHANG Shu-Ling,WANG Jun-Wei,HU Shi-Liang,WANG Tu-Tu,LI Shun-Chang,FAN Jia and SUN Jun-Zhi 10true Objective This study aims to explore the potential of different orders of magnitude single-nucleotide polymorphism (SNP) locus combinations for predicting distant kinship relationships. A high-density SNP locus set was constructed, and a comprehensive assessment of its inference capability was conducted.Methods Firstly, we selected three commercial chip panels, CGA (Chinese genotyping array, Illumina), GSA (Global screening array, Illumina), Affy (23MF_V2 high-density SNP array, Affymetrix) and merged them after quality control, forming a high-density SNP locus panel(1 180 k). Secondly, we selected 161 samples and collected their peripheral blood samples by using whole-genome sequencing technology. Within this sample population, the levels of kinship relationships fully covered the range from level 1 to level 9, and the number of kinship pairs at each level was consistently maintained at over 50 pairs. From 161 samples data of whole-genome sequencing, the 1 180 k locus set was extracted, which is referred to as the high-density SNP locus set in the following text. The kinship inference was conducted using the identity-by-descent (IBD) algorithm with the selected optimal parameters. To comprehensively evaluate the performance of the high-density SNP locus set in kinship inference, we compared it with the three commercial chip panels, the intersection of these three chip loci, and the control sets constructed by randomly reducing the number of the high-density SNP locus set. Based on the changes in the IBD lengths, as well as the dynamic trends in prediction accuracy, we conducted a scientific assessment of the kinship inference capability of the high-density SNP locus set.Results After screening, a set of 1 184 334 autosomal SNPs was obtained. During the process of screening the optimal IBD length threshold, the result revealed that 0 cM, 1 cM, and 2 cM all demonstrated good applicability. However, to avoid the issue of a large amount of redundant information caused by setting a too low IBD length threshold, this study ultimately selected 2 cM as the optimal threshold. Compared with the average results of three chip panels, the high-density SNP locus set increased the total IBD length and the average IBD length across levels 1-9; the accuracy of the confidence interval for level 8 was 70.97%, which represented a 3.50% improvement; the average confidence interval accuracy for levels 1-8 was 91.39%, representing a 1.00% increase; and the false negative rates at levels 8 and 9 were reduced by 2.42% and 6.76%, respectively. The system efficacy of the high-density SNP locus set for kinship inference of first to eighth degree relationships reached 98.91%. Through random reduction of the high-density SNP locus set results, it is found that increasing the number of SNPs with the panel, the detection efficiency of IBD length showed a significant upward trend. At the same time, the overall trend in the accuracy of kinship relationship prediction as well as the confidence interval accuracy also indicated that both metrics steadily increased with the addition of more loci.Conclusion The results show that the high-density SNPs panel significantly enhances the efficacy of distant kinship inference, accurately covering kinship degrees, with the average confidence interval accuracy for first to eighth degree relationships stably above 90%. The study finds that increasing the number of SNPs panel can improve the ability to predict distant kinship.]]> 2026/1/4 22:38:33 LI Jing,SUN Yi-Jie,ZHAO Wen-Ting,TANG Zi-Chen,LIU Jing and LI Cai-Xia 9true Objective Donkey hide is the sole legally designated raw material for the preparation of the traditional Chinese medicine Ejiao. The quality stability of donkey hide during preservation directly determines the efficacy and safety of Ejiao. This study focuses on the dynamic succession of microbial communities during the preservation of donkey hides from different origins, aiming to clarify the correlation between microbial biodiversity difference and the degradation profiles of hide collagen and critical biochemical components, thereby providing a theoretical foundation for developing targeted preservation strategies based on microbial regulation.Methods Donkey hides originating from four different regions were subjected to an accelerated microbial aging assay to simulate the spoilage process. The microbial community succession was analyzed using high-throughput sequencing. Microstructure changes and pore structure characteristics were assessed by scanning electron microscopy and mercury intrusion porosimetry, respectively. Additionally, the content of major components, including lipids, proteins, and sugars were determined by biochemical methods.Results After 96 h of aging, the collagen fiber structure in Africa donkey hides (ADH) exhibited significant degradation and collapse, followed by Xinjiang donkey hides (XDH). Instead, the microstructure of Dong’e black donkey hides (DDH) and Peru donkey hides (PDH) remained relatively intact. The porosities of DDH, XDH, PDH, and ADH increased from 27.9%, 15.7%, 30.3%, and 46.2% to 36.5%, 52.6%, 42.8%, and 57.7%, respectively, during the aging process, which suggested that the originally compact fiber structure was disrupted by microbial aging. Fourier transform infrared spectrometer analysis revealed the amide bands in XDH exhibited relatively weak intensity, and no collagen amide I band was observed in ADH. Meanwhile, the lipid and protein contents decreased in all four types of donkey hides, indicating that these components served as the primary nutrient sources for the growth of microorganism. Notably, the most severe collagen degradation was observed in XDH and ADH. A substantial increase was detected in the total soluble sugar in PDH aging solution and hydroxyproline in the ADH aging solution, respectively. These results indicated that donkey hides exhibit distinct patterns of structural degradation and nutrient utilization. Furthermore, the viable cells number of donkey hides increased sharply after 48 h of aging. Metagenomic analysis revealed that the relative abundance of Euryarchaeota in ADH, PDH and XDH declining from initial 93.19%, 97.73% and 30.08% to 0.79%, 1.43% and 0.02% after 96 h, respectively. Conversely, a significantly increase was observed in the abundance of Bacillota, with a marked increase in ADH, peaking at 92.75%. Additionally, the abundance of Pseudomonadota in PDH increased from 0.10% to 87.84%, suggesting that Bacillota and Pseudomonadota may be key factors exacerbating donkey hide spoilage. Unlike the other three types of donkey hides, the dominant bacterial phylum in DDH shifted from Pseudomonadota to Bacteroidota, characterized by a substantial abundance increase of Bacteroidota from 0.13% to 44.22%.Conclusion Regional variation in origin significantly influence the microbial aging of donkey hides, leading to distinct patterns of structural deterioration and differential nutrient utilization. Therefore, implementing origin-specific preservation strategies, through the precisely controlling environmental factors to suppress harmful phyla such as Bacillota and Pseudomonadota, is crucial for enhancing the storage quality of donkey hides.]]> 2025/12/22 14:46:59 ZHANG Meng,LI Qiu-Mei,KANG Jia-Wei,YU Jie,LI Xia and YU Yue 8true Objective Multiple sclerosis (MS) is a chronic inflammatory demyelinating disease of the central nervous system (CNS); however, its underlying neurological pathogenic mechanisms remain incompletely understood. Endogenous formaldehyde (FA), a metabolic byproduct of methylation-demethylation cycles, has recently been implicated in neurotoxicity, oxidative damage, and cognitive impairment. This study aimed to investigate whether excessive FA contributes to myelin sheath demyelination in mice and to evaluate the protective effects and mechanisms of two FA-elimination strategies: sodium bisulfite (NaHSO3), a classical FA scavenger, and polyethylene glycol-modified astaxanthin nanoparticles (PEG-ATX@NPs), a brain-targeted nano-antioxidant formulation.Methods A chronic demyelination model was established by feeding female C57BL/6J mice a diet containing 0.2% cuprizone (CPZ) for four weeks, followed by a two-week intervention period. Eighty mice were randomly assigned to four groups: NS (normal saline), CPZ+NS, CPZ+NaHSO3, and CPZ+PEG-ATX@NPs. Behavioral tests, including open-field, Y-maze, and pole-climbing assays, were conducted to assess locomotor activity, motor coordination, and working memory. FA levels in serum, corpus callosum, and spinal cord were measured using an Na-FA fluorescent probe and quantified via in vivo and ex vivo fluorescence imaging. Neuroinflammatory responses were evaluated by measuring TNF-α, IL-1β, and IL-6 levels using ELISA, while oxidative stress was assessed by reactive oxygen species (ROS) fluorescence intensity. Demyelination was examined via Luxol fast blue staining, and microglial activation was analyzed by Iba1 immunofluorescence. Correlation analyses were performed to explore relationships among FA levels, inflammatory cytokines, ROS intensity, and behavioral parameters.Results Compared with the NS group, mice in the CPZ+NS group exhibited significant weight loss, impaired motor coordination and memory, and markedly reduced myelin regeneration (P<0.05). FA levels and pro-inflammatory cytokines were significantly elevated in serum, corpus callosum, and spinal cord (P<0.05). FA-associated fluorescence in brain and spinal tissues, as well as ROS intensity across all tissues examined, also increased substantially (P<0.05). CPZ treatment induced pronounced microglial activation and severe demyelination in the corpus callosum (P<0.01). Both NaHSO3 and PEG-ATX@NPs effectively reduced FA accumulation in the brain and spinal cord, attenuated demyelination, suppressed microglial activation, decreased inflammatory cytokine levels, and improved motor and cognitive performance. These results confirm that CPZ induced severe demyelination accompanied by oxidative stress, neuroinflammation, and abnormal FA accumulation. Following intervention with either NaHSO3 or PEG-ATX@NPs, endogenous FA levels in the CNS were substantially reduced. Both treatments alleviated demyelination and significantly decreased the number of activated microglia. Levels of TNF-α, IL-1β, and IL-6 in serum, corpus callosum, and spinal cord were downregulated. Behavioral performance improved significantly, as evidenced by enhanced locomotor activity, better coordination, and improved memory function. These findings indicate that both FA-scavenging agents mitigate CPZ-induced biochemical and behavioral abnormalities.Conclusion This study demonstrates that excessive endogenous FA is closely associated with cognitive impairment, inflammatory dysregulation, and demyelination in a CPZ-induced chronic demyelination mouse model. Clearing abnormally elevated FA effectively reduces neuroinflammation, suppresses microglial overactivation, decreases oxidative stress, and alleviates demyelination, ultimately improving motor and cognitive outcomes in mice. These results suggest that targeting endogenous FA represents a promising therapeutic strategy for MS and other demyelinating disorders. Further investigations are warranted to explore the long-term safety, dosage optimization, and molecular pathways involved in FA-mediated neurotoxicity.]]> 2025/12/29 0:00:00 Lü Wan-Jia,ZENG Xin,TONG Zhi-Qian,XING Yang,YANG Xu,WU Mei-Na and MA Ping 7true Objective The accuracy of Y-chromosome haplogroup assignment is crucial for tracing paternal lineage in male samples. With the advancement of high-throughput sequencing technologies, high-density Y-SNP genotyping from whole-genome or array-based data has become a standard method for determining Y-chromosome haplogroups. This study systematically evaluated the performance of 4 commonly used high-density SNP genotyping systems—namely, the Global Screening Array (GSA), Chinese Genotyping Array (CGA), Affymetrix array, and the 1240K capture panel—for haplogroup assignment. This work provides a reference for data comparison across different systems.Methods We extracted genotype data for the 4 Y-SNP panels from 30× whole-genome sequencing (WGS) data of 1 590 male samples from the 1000 Genomes Project. Additionally, GSA array genotype data from 384 relative pairs (spanning 1st- to 12th-degree relationships) from 109 Chinese Han families were collected. Haplogroup assignment was performed using Y-LineageTracker v1.3.0 software. We assessed the concordance and resolution of haplogroup assignments between the four Y-SNP panels and the WGS data. The consistency and resolution of haplogroup assignments were also evaluated for both the 1000 Genomes Project samples and the 109 family samples collected in this study. Furthermore, the impact of varying numbers of Y-SNPs on haplogroup assignment was examined.Results The GSA and CGA panels demonstrated superior resolution and discrimination of haplogroup subclades compared with the other two panels. The haplogroup assignments from the GSA, CGA, and 1240K panels showed high concordance with WGS data, with consistency rates exceeding 88.70%, whereas the Affymetrix platform exhibited a significantly lower consistency rate of 61.89%. Specifically, the GSA and CGA panels consistently demonstrated superior performance compared with the other two panels in the assignment of haplogroups O-M175 and H-L901, achieving complete concordance (100%) for both haplogroups. In contrast, the Affymetrix panel erroneously assigned all individuals belonging to haplogroup O-M175 to haplogroup K2-M526. Furthermore, its accuracy for haplogroup H-L901 was exceedingly low, at merely 1.41%. This poor performance was characterized by the misassignment of 98.59% of H-L901 samples—specifically, 1.41% to J-M304 and a predominant 97.18% to F-M89. For haplogroup R-M207, all four panels exhibited uniformly high levels of consistency, with concordance values exceeding 94.00%. Notably, for haplogroup E-M96, the 1240K and Affymetrix panels outperformed the GSA and CGA panels in terms of concordance, representing the first instance in which these two panels surpassed the latter. Conversely, for haplogroups J-M304, Q-M242, and I-M170, all 4 panels showed relatively elevated misclassification rates, with the Affymetrix array demonstrating the poorest overall performance. None of the four panels showed any discordant haplogroup assignments among the familial relative pairs analyzed. A positive correlation was observed between the number of Y-SNPs (ranging from 1 000 to 10 000) and classification consistency; however, classification consistency plateaued when the number of Y-SNPs exceeded 10 000. Furthermore, a random sampling analysis conducted on the GSA and CGA panels demonstrated that the haplogroup misclassification rate exhibited negligible fluctuation across the Y-SNP range of 500 to 1 000. Conversely, a marked enhancement in classification consistency was observed as the number of markers increased from 1 000 to 5 000, ultimately reaching a plateau within the interval of 5 000 to 8 000 markers.Conclusion These findings indicate that the GSA and CGA panels provide high resolution and concordance, delivering reliable Y-haplogroup assignment for forensic investigations.]]> 2025/12/26 11:45:39 ZHANG De-Qin,WANG Chun-Nian,LOU Lin-Lin,NI Meng,GAO Jing,HUANG Jiang and JIANG Li 6true Objective This Study was conducted to investigate the interaction mechemisms between gut microbiota dysregulation and ischemic stroke by establishing a rat model of ischemic stroke and employing fecal microbiota transplantation (FMT).Methods A preliminary experiment was conducted to establish an antibiotic-induced pseudo-sterile (ABX) rat model through antibiotic treatment, and a cerebral ischemia model was prepared using the middle cerebral artery occlusion (MCAO) method. Fecal microbiota from stroke patients and healthy individuals were transplanted via FMT, followed by behavioral testing. 16S rRNA sequencing was used to analyze the microbial community, hematoxylin and eosin (HE) staining to observe histopathological status, transmission electron microscopy (TEM) to examine the tight junction structure of the small intestine, and enzyme-linked immunosorbent assay (ELISA) to detect levels of inflammatory factors and intestinal barrier-related markers.Results 16S rRNA sequencing of fecal samples showed that compared with the normal control group and the metronidazole group, the abundance and diversity of fecal microorganisms in the quadruple antibiotic group were significantly reduced, indicating successful establishment of the ABX model. After transplanting fecal microbiota from stroke patients into ABX rats, significant changes in gut microbiota composition were observed. Behavioral tests revealed that the MCAO model group showed significant decreases in both horizontal movement and vertical exploration abilities. ELISA results indicated that IL-17 concentration in the ABX+mFMT (antibiotic-treated+control fecal microbiota transplantation) group was lower than in the ABX+cFMT (antibiotic-treated+model fecal microbiota transplantation) group, suggesting that IL-17 may serve as a key inflammatory indicator for evaluating the impact of stroke intervention on gut microbiota. Triphenyltetrazolium chloricle staining (TTC) staining suggested that gut microbiota intervention may increase the risk of stroke. HE staining showed that, except for the control group, all groups exhibited ischemic changes and inflammatory infiltration in brain tissues. TEM revealed that microvilli of small intestinal epithelial cells in the ABX+mFMT group were sparser than those in the ABX+cFMT group, indicating that microbial intervention affects intestinal barrier function.Conclusion The ABX model established using broad-spectrum antibiotics showed no significant differences in physiological characteristics compared to normal rats, and the findings were consistent with those from germ-free rat models. Stroke prognosis appears to be influenced by intestinal dysbiosis, accompanied by significantly elevated levels of the pro-inflammatory cytokine IL-17, which may exacerbate neural injury via the gut-brain axis. Behavioral experiments indicated that transplantation of gut microbiota from stroke rats impaired cognitive function. Furthermore, IL-17 demonstrated sensitivity to alterations in the gut microbiota, suggesting its potential as a key therapeutic target for stroke intervention.]]> 2025/12/30 14:23:35 WANG Ting,ZHANG Jing-Hao and JIANG Chao 5true Objective Cleft palate (CP) is a common congenital deformity often associated with velopharyngeal insufficiency (VPI), which disrupts the physiological coupling between respiration and speech. Conventional clinical assessments, such as nasometry and spirometry, provide limited static data and fail to visualize the dynamic spatiotemporal distribution of lung ventilation during phonation. This study introduces spatiotemporal electrical impedance tomography (ST-EIT) to evaluate speech-respiratory functional features in CP patients compared to normal controls (NC). The aim is to characterize multi-domain respiratory patterns and to validate an interpretable machine learning framework for providing objective, quantitative evidence for clinical assessment.Methods Seventy-five participants were enrolled in this study, comprising 37 patients with surgically repaired CP and 38 healthy volunteers matched for age, gender, and body mass index (BMI). All subjects performed standardized sustained phonation tasks while undergoing synchronous monitoring with a 16-electrode EIT system and a pneumotachograph. A comprehensive feature engineering pipeline was developed to extract physiological parameters across 3 complementary domains. (1) Temporal domain: including inspiratory/expiratory phase duration (tPhase), time constants (Tau), and inspiratory-to-expiratory time ratios (TI/TE); (2) airflow domain: comprising mean flow, peak flow, and instantaneous flow at 25%, 50%, and 75% of tidal volume; and (3) spatial domain: quantifying global and regional tidal impedance variation (TIV), global inhomogeneity (GI), and center of ventilation (CoV). Extreme Gradient Boosting (XGBoost) classifiers were trained using 5 distinct data sources (Spirometry, Nasometry, Inspiratory-EIT, Expiratory-EIT, and fused ST-EIT). Model performance was rigorously evaluated via stratified 5-fold cross-validation, and Shapley additive explanations (SHAP) were employed to quantify global and local feature contributions.Results The CP group exhibited a distinct respiratory phenotype compared to controls. In the temporal domain, CP patients showed significantly shorter inspiratory (1.60 s vs. 1.85 s, P<0.001) and expiratory phase durations (2.45 s vs. 3.95 s, P<0.001), indicating a rapid, shallow breathing rhythm. In the airflow domain, while inspiratory flows were comparable, the CP group demonstrated significantly elevated mean and peak flows during the expiratory phase (P<0.001), reflecting compensatory respiratory effort. Spatially, CP patients presented significant ventilation redistribution, characterized by higher regional TIV in the right-anterior (ROI1) and left-posterior (ROI4) quadrants, but lower TIV in the left-anterior (ROI2) quadrant. In terms of diagnostic accuracy, the multi-modal ST-EIT model achieved the highest performance (AUC: 0.915±0.012, Accuracy: 0.843±0.019, F1-score: 0.872±0.017), substantially outperforming models based on spirometry (AUC: 0.721) or nasometry (AUC: 0.625) alone. Interpretability analysis revealed that spatial domain features were the most critical, contributing 53.4% to the model’s decision-making, followed by temporal (25.0%) and airflow (21.6%) features.Conclusion ST-EIT successfully captures the temporal, airflow, and spatial deviations in CP speech respiration that are undetectable by conventional methods—specifically, rapid phase transitions, hyperdynamic expiratory airflow, and regional ventilation heterogeneity. This study validates ST-EIT as a robust, non-invasive, and radiation-free tool for characterizing speech-respiratory dysfunction, offering high clinical value for bedside screening, rehabilitation planning, and longitudinal monitoring of patients with cleft palate.]]> 2026/1/12 16:53:26 WU Yang,ZHANG Xiao-Jing,YU Hao,JIANG Cheng-Hui,SUN Bo and YAO Jia-Feng 4true Objective Pioneer transcription factors (PTFs) possess the unique ability to recognize and bind their target DNA sequences within compacted nucleosomal DNA, thereby initiating chromatin opening and gene expression. They play pivotal roles in fundamental biological processes such as embryonic development, cellular reprogramming, and tumorigenesis. The specific regulatory mechanism by which nucleosomal rotational positioning governs PTF-nucleosome interactions remains inadequately elucidated. This study aims to systematically investigate the role of the rotational orientation of motifs in PTF-nucleosome binding.Methods We employed a DNA deformation energy model to predict the rotational positioning of DNA on nucleosomes. We analyzed high-throughput in vitro data from the NCAP-SELEX assay, which profiles the binding landscapes of numerous transcription factors to nucleosomal DNA. For in vivo analysis, we integrated genome-wide binding data (ChIP-seq) and nucleosome positioning data (MNase-seq) for eight well-characterized pioneer factors (OCT4, SOX2, KLF4, GATA4, MYOD1, FOXA1, CEBPA, and ASCL1) in human cells. Binding motifs were classified as “TF-bound” if they overlapped with ChIP-seq peaks and “TF-unbound” otherwise. DNA bendability profiles and fast Fourier transform (FFT) analysis were used to assess rotational positioning patterns around these motif sites. This analytical framework was further applied to specific biological contexts, including cellular reprogramming from IMR90 fibroblasts to induced pluripotent stem cells (iPSCs) and the differentiation of human embryonic stem cells (hESCs) to human neuroectodermal cells (hNECs).Results Our in vitro analysis revealed a strong dependence of transcription factor binding on the rotational orientation of TF-binding motifs. For SOX7, the unbound motifs at specific enrichment peaks exhibited a rotational phase clearly opposite to that of the SOX7-bound motifs. Similarly, analysis of P53 binding sequences confirmed that successful binding in vitro correlated with model-predicted exposure of the DNA minor groove at the motif center, consistent with P53’s binding mode. Genome-wide in vivo analysis of the eight PTFs showed that their DNA binding motifs were generally associated with DNA sequences exhibiting significant 10-bp periodicity in bendability, suggesting an inherent potential for nucleosome association. Crucially, for most factors (except ASCL1), the average rotational positioning preferences were remarkably similar between TF-bound and TF-unbound motifs. This indicates that, at a global genomic level, rotational positioning is not the primary determinant dictating whether a nucleosomal motif is bound by its cognate PTF in vivo. This phenomenon persisted during cellular reprogramming (IMR90 to iPSC), where the rotational positioning of OSKM factor motifs bound versus unbound in nucleosomal regions showed no significant overall difference. Interestingly, during hESC differentiation to hNECs, SOX2 binding sites underwent comprehensive reprogramming. In hNECs, the rotational positioning of nucleosomal SOX2-bound motifs was significantly different and, unexpectedly, opposite to the general preference observed in hESCs and for unbound motifs in hNECs, suggesting a cell context-dependent rewiring of binding mechanisms.Conclusion This study suggests a distinction in the role of DNA rotational positioning in TF-nucleosome binding between in vitro and in vivo environments. While rotational positioning critically governs the binding efficiency of factors like SOX7 and P53 in simplified in vitro systems, PTFs in vivo appear to overcome this steric hindrance at the binding interface. The ability of PTFs to bind nucleosomal motifs, even when key interaction surfaces are partially buried, might stem from their unique structural properties (e.g., intrinsically disordered regions, DNA distortion/binding domains), nucleosome breathing which transiently exposes DNA, and potential cooperativity with other factors. Our results highlight the unique capacity of pioneer factors to drive chromatin openness through mechanisms beyond rotational positioning.]]> 2025/12/4 14:50:58 LIU Guo-Qing,GUO Xing-Yue,CANG Jing,ZHANG Zhi and LIU Guo-Jun 3true Objective With the continuous evolution of severe acute respiratory syndromes-coronary virus 2 (SARS-CoV-2) Omicron subvariants, particularly the emergence of BA.2.86 and its descendant JN.1, the efficacy of current neutralizing antibodies has faced substantial challenges. The JN.1 variant, noted for its pronounced immune evasion capacity, has rapidly become the globally dominant strain. Elucidating its escape mechanisms is therefore essential to guide the development of next-generation broad-spectrum vaccines and neutralizing antibody therapeutics. This study aimed to investigate the immune evasion mechanisms of JN.1 against broadly neutralizing antibodies, focusing on the effects of key receptor-binding domain (RBD) mutations on antibody binding and neutralization, thereby providing theoretical support for countering ongoing viral evolution.Methods We employed a multidisciplinary approach to systematically assess the binding and neutralizing activities of three broad-spectrum neutralizing antibodies (XGv074, XGv302, and XGv303) against BA.2.86 and JN.1. Binding affinities (KD values) of antibodies to variant RBDs were determined using bio-layer interferometry (BLI). Cryo-electron microscopy (cryo-EM) was used to resolve the structure of the BA.2.86 Spike trimer in complex with antibody antigen-binding fragments (Fabs), achieving a resolution of 3.47 ? for the BA.2.86 S-trimer bound to XGv302. Molecular dynamics simulations and binding free-energy decomposition were conducted to quantify the contributions of key mutations at the antibody-RBD interface. Additionally, sequence alignment and structural modeling were performed to evaluate the role of conformational flexibility in the antibody heavy-chain complementarity-determining region 3 (HCDR3) in mediating tolerance to mutations.Results Experimental data showed that XGv074, XGv302, and XGv303 retained neutralizing activity against BA.2.86 but exhibited markedly reduced binding to JN.1, with only XGv074 maintaining weak neutralization (IC50=2.3 mg/L). Cryo-EM structures revealed that all three antibodies targeted the RBD tip, overlapping with the ACE2-binding region. The JN.1-specific L455S mutation disrupted the hydrophobic interaction network between XGv302 and the RBD (involving key residues such as Y421 and L455), resulting in complete loss of neutralization. Binding free-energy decomposition further identified L455 and Y421 as energetic hotspots (ΔG<-3 kcal/mol), with the L455S mutation directly impairing antibody binding. XGv074, owing to greater conformational flexibility in its HCDR3 region, partially tolerated the mutation and retained weak binding. Molecular dynamics simulations showed that the L455S mutation not only eliminated the energetic contribution of this residue but also caused a concurrent decrease in binding free energy of neighboring residues, thereby reducing overall interface stability.Conclusion The JN.1 variant escapes broad-spectrum neutralizing antibodies primarily through the L455S mutation in the RBD, which disrupts energetic hotspots and remodels the antibody-binding interface. Antibody conformational flexibility enhances adaptability to such mutations, providing new insights for broad-spectrum antibody design. These findings highlight the critical roles of epitope energy distribution and antibody flexibility in maintaining neutralization breadth, offering essential guidance for the rational design of next-generation vaccines and antibody therapeutics: specifically targeting conserved energetic hotspots while enhancing CDR flexibility to counter immune evasion driven by viral evolution.]]> 2025/8/20 14:57:08 XIE Jia-Wen,LIU Tian-Ci,GUO Meng-Tian,FENG Lu-Lu,SUN Ming-Chen,LIU Pan and ZHU Qian-Hui 2true Objective This study aimed to construct a cell co-culture microfluidic chip based on droplet microfluidic to investigate the influence of multicellular interactions in complex microenvironments on the sensitivity of anti-tumor drugs.Methods We constructed a droplet microfluidic chip consisting of 12 co-culture units, with each unit containing 4 microwells for holding cell droplets, enabling the co-culture of 4 types of cells. To evaluate whether the co-culture of multiple cell types can be achieved in the droplet microfluidic chip, as well as to observe and analyze the interactions between different cell types, we investigated the interaction between microenvironmental cells and tumor cells through cell co-culture experiments. To construct a stable co-culture system capable of evaluating drug sensitivity, we conducted diffusion experiments with blue ink and model drugs to investigate the ability of drugs to diffuse in the chip and be taken up by cells. To investigate the effect of the complex microenvironment on cellular drug sensitivity, we carried out cell co-culture experiments combined with drug treatment to explore the changes in the drug sensitivity of tumor cells in the presence of microenvironmental cells. To explore the reasons for drug resistance in tumor cells under co-culture conditions, we detected DNA double-strand break marker using immunofluorescence.Results Experiments on cell culture within droplets showed that cells in each droplet exhibited good proliferation ability and consistent cell status, laying a foundation for the co-culture of multiple kind of cells. Cell co-culture experiments showed that compared with the mono-culture group, the numbers of LoVo cells, HUVECs, and macrophages in the co-culture group increased significantly. This confirms that there are obvious interactions between cancer-associated fibroblasts (CAFs), endothelial cells (HUVECs), macrophages, and tumor cells in the microenvironment, which promotes cell proliferation in the co-culture chip. Experiments on blue ink diffusion showed that drugs could diffuse uniformly and effectively into wells in different directions within the chip. Experiments on the diffusion of doxorubicin (a model drug) demonstrated that identical cells in different wells exhibited consistent drug uptake capacity. Additionally, cell co-culture experiments combined with oxiliplatin treatment revealed that with the concentration of 80 μmol/L, the survival rate of LoVo cells cultured alone was only 25%, whereas it reached 96% under co-culture conditions; With the concentration of 160 μmol/L, the survival rate of LoVo cells cultured alone was merely 2%, while that under co-culture conditions was 50%. These results indicate that the complex microenvironment composed of CAFs, HUVECs, and macrophages significantly reduces the drug sensitivity of LoVo cells to oxaliplatin through intercellular interactions. The immunofluorescence results showed that the expression level of γH2AX in LoVo cells decreased under co-culture conditions.Conclusion Our study achieved co-culture of the main constituent cells of the tumor microenvironment and analysis of their drug sensitivity in a droplet microfluidic chip for the first time. The research found that crosstalk between different microenvironmental cells strongly affects the drug sensitivity of tumor cells to oxaliplatin, suggesting that targeting the interactions between tumor microenvironmental cells is an effective strategy to improve the efficacy of tumor therapy. Our study provides new methods and approaches for the efficacy evaluation of anti-tumor drugs and the screening of new drugs. In addition, the open structural design of the co-culture chip can be combined with various omics technologies to analyze the molecular characteristics of cells under co-culture and drug treatment conditions. This is expected to provide new methods and experimental evidence for elucidating the mechanisms of drug action and identifying novel drug targets in the context of the microenvironment.]]> 2025/12/17 11:41:25 ZHANG Xue-Tong,CHEN Shuo and FANG Jin 1true