Abstract:Synucleins, as a family of small proteins enriched in presynaptic element in CNS, share a secondary structure feature of periodicity of amphipathic α-helix with 11 amino acid residues throughout the N-terminus. The physiological function of synucleins is unknown, but large quantities of research suggest that synucleins may be implicated in synaptic genesis and plasticity. Also, the relations of synuleins to many neurodegenerative diseases such as Alzheimer's and Parkinson's as well as to the invasion of breast cancer have been gaining attention of researchers.

Abstract:Insulin-like growth factorⅠ(IGF-Ⅰ) and insulin are two important members in the insulin family with highly homologous structure. They bind to the homologous receptor and produce similar physiological function, but the major function is different. The major function of IGF-Ⅰ is growth-promoting whereas insulin plays a key role in the glucose uptake and metabolism. The bases of the physiological function and its expression of a protein are the molecular structure of the protein and the molecules involved in the expression of the function, such as receptors and signal molecules, etc. The recent progresses in the studies on the structural basis and molecular basis of physiological function of IGF-Ⅰ and insulin are reviewed.

Abstract:The role of the Epstein-Barr Virus-encoded LMP-1 in mediating new signal transduction pathway becomes more attractive than ever.The structural properties,and function of LMP-1,the effects of TRAF/TRADD,the activation of NF-B,AP-1,and JAK/STAT were reviewed.

Abstract:DNA dependent protein kinase（DNA-PK）is a DNA ends binding protein composed of Ku protein and DNA-PKcs.Ku protein can bind directly to DNA ends，which will stimulate DNA-PK catalytic subunit.DNA-PK is a very important cellular factor which plays important role in the multiple cellular processes such as DNA repair，gene recombination，DNA replication and transcription.

Abstract:bcl-x gene is an important member of the bcl-2 multigene family.Three isoforms:Bcl-X_L,Bcl-X_S and Bcl-X_γ,are generated by alternative splicing.Bcl-X_L and Bcl-X_γ inhibit apoptosis. However,Bcl-X_S enhances apoptosis. Bcl-x plays a role in cell development,homeostatic balance, tumorigenesis and prognosis, which regulates apoptosis through interacting with Bcl-2, Bax, Bad and so on.

Abstract:Apoptosis is a highly organized mechanism by which cells undergo programed cell death. Apoptosis participates in many physiological and pathological processes. Recent experimental approaches suggest that sphingolipid metabolites participate in key events of apoptosis. Being a lipid second messenger, sphingomyelin lysis product-ceramide play an important role in inducing and preventing apoptosis.

Abstract:In recent years, the effects of the N-methyl-D-aspartate(NMDA) receptor on the long-term potentiation(LTP) and the intracellular cascade reaction after the activation of the NMDA receptor are emphasised widely by people through antagonists and gene knockout technique. These studies show the processeing of induction and maintenance of the LTP. The results provide evidences for the pre- and post-synaptic mechanism of the LTP.

Abstract:Multicellular morphogenesis is the main content of myxobacterial social behavior. The morphogenesis contains aggregation of cells under nutrient deprivation, autolysis, development of fruiting bodies and formation of myxospores. There is a complicated system of signals and regulation during morphogenesis, which is much more similar to eukaryotes. Myxobacterium is an important model for studying cellular differentiation and development in prokaryotes and biological evolution.

Abstract:Alzheimer's disease(AD) is a neurodegenerative disorder.It is the most common cause of dementia in aged population and also a main cause of death in aged people.The oxidative stress hypothesis of AD has focused on and described from four aspects:Molecular basis and oxidative stress basis of AD pathogenesis,and β amyloid aggregation and cytotoxicity related to reactive oxygen species.

Abstract:Yeast three-hybrid system based on yeast two-hybrid system can be used in research of the interaction among three kinds of proteins and interaction among the proteins and RNA or interaction between protein and drug.Its theory,application and shortcoming were summarized.

Abstract:HSP70 molecular chaperone participate in various cellular processes under normal and stress conditions, including the folding of nascent polypeptides, assembly and disassembly of multimeric protein structures, membrane translocation of secreted proteins and protein degradation. All of these activities rely on the ATP-regulated association of HSP70 with short hydrophobic segments in substrate polypeptides. Significant progress has been made in the understanding of the crystal structure of the C-terminal polypeptide-binding domain and the ATP-dependent mechanisms of HSP70.

Abstract:Using DIG (digoxigenin) labeled GAP-43 cDNA as probe, the 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.

Abstract:To explore the DNA binding proteins of mouse mbcl-2 regulatory region in the apoptotic process of mouse fibroblast cell line (C3H 10 T1/2 Cl 8) and its transformed counter part, the mouse bcl-2 (mbcl-2) regulatory region was amplified by PCR. The PCR products were subcloned into pGEM-T vector system and then checked by sequencing. Using the product as probe, DNA binding proteins of mbcl-2′s regulatory region in nuclear protein extracts of both cell lines were studied. Southwestern blotting results showed that a 53 ku protein binds with the probe.Its binding signal strengthened after exposing both cells to 5-Fu for 12 h, as another DNA binding protein (100 ku) also binds with the probe though its binding signal weakened after 5-Fu treatment. The results suggested that p53 protein may be the negative regulatory factor of mbcl-2 gene. An unidentified 100 ku protein may be the positive regulatory factor of mbcl-2 gene.

Abstract:To detect cleavage activity of anti-HBV two-unit ribozyme and single ribozyme,and compare two conected ribozyme cleavage efficiency with two mixed ribozymes, firstly, Rz1,Rz3 and Rz13 ribozymes transcription vectors were constructed,then cleavage activity of Rz1,Rz3 and Rz13 ribozymes on target RNA were observed. The results showed that antiviral activity of two-unit ribozyme,whether connected (Rz13)or two mixed ribozymes, has higher cleavage efficiency than single ribozyme. Cleavage efficiency has no difference between connected (Rz13) and two mixed ribozymes(n=2,P＞0.05).

Abstract:A eukaryotic vector highly expressing PKC_γ,was constructed by DNA recombination and transfected into C₃H₁₀T_1/2 cells by gene transfection. The NCP4 cells stably expressing PKC_γ 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 PKC_γ 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 PKC_γ level directly affected the increase of growth rate and lead to some transforming phenotypes in C₃H₁₀T_1/2 cells.

Abstract:Prephosphorylation of τ by cAMP-dependent protein kinase (PKA) significantly stimulated its consecutive phosphorylation catalyzed by glycogen synthetase kinase-3 (GSK-3). After digestion of phosphorylated τ with trypsin, 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.

Abstract:Atherosclerosis(As)rabbit model was developed by feeding with high-cholesterol diet for 12 weeks,then As rabbits were injected intravenously with human plasma HDL preparation for 10 weeks,the effect of HDL on activity of LDL receptors on As rabbit liver plasma membranes was investigated. It was shown that the value of B_max of LDL receptor was significantly lower in As rabbit than that in normal rabbit(P＜0.01), while the value of K_d showed no difference; in HDL-treated rabbits, the value of B_max increased significantly compared with the rabbits without HDL treatment(P＜0.01), while the value of K_d 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.

Abstract:In order to observe the difference in gene expression in the kidney of SHR and WKY rats, mRNA differential display was undertaken to screen differently expressed genes in the kidney of SHR and WKY rats aged 12 weeks by using T11G and AP6 primer. One cDNA fragment was identified in the kidney of SHR and was selected as interesting band. The techniques of T-A cloning, Northern hybridization, 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.

Abstract:Expression of active horseradish(Armoracia rusticana) peroxidase (HRP) in microbes is necessary not only to gain an insight into its mechanisms, structure and function, and physiological role in plants, but also to supply large quantities of this useful enzyme. Exploiting new approach to express HRP in methyltrophic yeast, 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.

Abstract:Acetylcholine localization was studied immunohistochemically in neurons of the area 17 and 18 in the hamster visual cortex using anti-acetylcholine serum. It was found that the Ach-labeled cells are nonpyramidal neurons primarily and a few pyramidal neurons also exist. These neurons were found to be distributed in layer Ⅱ～Ⅵ, concentrated in the layer Ⅱ～Ⅳ. The neurons in the area 17 having greater density are arranged to form a columns perpendicular to the surface of the visual cortex and the neurons in the area 18 having smaller density are distributed dispersely. In the different layers of the visual cortex, the type of stained neurons has variation with that the majority of these neurons were bipolar in layer Ⅱ, Ⅳ and the multipolar cells in the other layers.

Abstract:Human plasminogen activator inhibitor 1 (PAI-1) gene was recombined into a replication-defective adenovirus vector AdCMVHSgD, co-transfected with plasmid pJM17 into human embryo kidney cell line 293 cells. Recombinant virus was produced without use of agarose overlay, identified by PCR analysis and used to infect B16 (F10) cells. The inhibition of plasminogen activator (PA) activity of PAI-1 was revealed in the elute and supernatant of infected B16 cells by regular and reverse agarose-fibrin autography.

Abstract:The routine iodination methods are easy to lead to biological inactivation of cytokines, which is one of reasons for failure of ligand receptor binding assay. By using two-phase iodination of chromine-T, rhG-CSF and rhEPO were iodinated with Na¹²⁵I and the characteristic of EPO receptor in BET-2 cell and G-CSF receptor in NFS-60 cell were observed by receptor ligand binding assay. It was showed that the radioactivity of ¹²⁵I-G-CSF and ¹²⁵I-EPO were much higher than that obtained by other methods and it was found that there are high and low affinity EPO receptors in BET-2 cells and one high affinity G-CSF receptor in NFS-60 cells. These results are in accord with literature data. It suggests that the two-phase iodination of chromine-T is an ideal method to iodinate cytokines.

Abstract:A method for purifying HA from ox eyes was studied.Using 0.125 mol/L Na₂SO₄ other than 0.1 mol/L NaCl to dissolve the semifinished product, Two HA forms in Na₂SO₄'s solution were found. One in staple could be absorbed by CBP, and the other in powder could be returned by fractional dilution afterwards. In 0.1 mol/L NaCl solution, HA could only be recovered by CBP absorption. It displayed staple. The recoverable rate of the method was 1.7 times than others.The physical and chemical characters of the product were analysed. It was proved that higher purity recovery and lower cost than other methods were gained.

Abstract:A nested PCR was used to amplify the rare target sequence and 5% formamide was used to eliminate the nonspecific amplification. These modifications make clone the flanking sequence adjacent to the known sequence of human genome very effective and highly specific.

Abstract:To investigate the gene expression of growth factor Wnt-5A in different phases during cell cycle at transcriptional level.To synchronize the renal cell carcinoma GRC-1 cell line was by double thymidine blocks and high-pressure N2O gas methods. Using Semi-quantitative RT-PCR (reverse transcriptase polymerase chain reaction) to amplify the cDNAs of Wnt-5A in different phases. Result showed that the mRNA expression of growth factor Wnt-5A was detected in RCC GRC-1cell line. In S phase, the highest level of Wnt-5A was observed, medial and lowest was in G1 and M phase, respectively. The differences between S and M stages were significant (P＜0.05).Therefore growth factor Wnt-5A has the potential effect on tumorigenesis, It contributes to all phases during cell cycle, but gives more influence to S phase during cell cycle.

Abstract:The proliferation of arterial SMC plays an important role in process of atherosclerosis. Oxidized HDL(OX-HDL) can increase ³H-TdR incorporation to DNA of cultured human arterial SMC, so promote proliferation of SMC. A study on effect of OX-HDL on proliferation of cultured human arterial SMC with MTT method was made. It was showed that native HDL(N-HDL) has no effect on proliferation of SMC but OX-HDL significantly stimulated the proliferation of SMC (P＜0.01);N-HDL significantly inhibited proliferation of human SMC induced by OX-LDL (P＜0.01) and OX-HDL significantly enhanced proliferation of human SMC induced by OX-LDL (P＜0.01).

Abstract:In order to study the mechanism of enzyme-linked immunoassay for octapeptide cholecystokinin（CCK-8）,ELISA procedures were based on the pretreatment that polystyrene microplates were irradiated by ultraviolet and activated by glutaraldehyde prior to coating,and the binding nature of CCK-8 to microplates was revealed by different washing methods including dH₂O,PBS-T and SDS. The results showed that this pretreatment led to the covalent attachment of CCK-8 to the solid-phase steadily and fit to different sources of microplates. The treated microplates could be stored at least 4 weeks not to influence their ability of binding antigens.The intra- and inter- variation coefficients were 4.75% and 7.80% respectively.This polypeptide ELISA was characterized by covalent attachment between antigen and solid-phase carrier and was proved to be highly stable,reproducable and available.

Abstract:Ultra-weak chemiluminescence analytical technology have a particular excellence in that determine the types of antioxidant functions and the antioxidant activity of the drugs. The methods and results that to determine antioxidation of several drug using BPCL analyzer was introduced. It is approved that the technology and analyzer were useful in the practical fields. The principle performance of data acquisition and analysis by BPCL was described.

Abstract:The serum cardiac enzymes in 30 cases of patients with acute myocardiac infarction(AMI) were studied. From the onset to 48 hours of AMI, the sera were collected in three different stages. By comparing with electrocardiogram respectively, the activities of AST,CK, LDH and CK-MB were measured. The results revealed that the highest positive rate of serum cardiac enzymes were 90%(AST,CK-MB), and the lowest positive rate was 70%(LDH).By using Q wave as a confirmatory index of AMI, the negative rate of cardiac enzymes was 40%. The early detection of increasing cardiac enzymes could indicate the presence of AMI. An increasing of CK-MB in patients with AMI indicated a possible new infarction occurred. A patient had increasing of cardiac enzymes within 6 hours of AMI might have a poor prognosis.

Abstract:The Chinese nomenclature of “prion” and “ribozyme” is discussed.The existing Chinese terms for these two kinds of molecules are not able to reflect the accurate characters of them.A new rule of Chinese nomenclature for the prion and ribozyme is suggested.

Abstract: