Abstract:Nonalcoholic fatty liver disease (NAFLD) is a common chronic liver disease with an increasing morbidity. However, its pathogenesis is still elusive and its diagnosis and treatment need to be improved. The emergence of proteomics has pushed NAFLD research forward and twenty-one associated studies have been reported by now. Recent progress in proteomic techniques allows evaluation of molecular changes associated with disease, thereby permitting to identify novel biomarkers and therapeutic targets. Herein, the application of proteomics in the study of the diagnosis, pathogenesis and other related fields of NAFLD were comprehensively reviewed. Firstly, empirical knowledge about categories of subjects and samples, experimental methods and biomarker selection was summarized. Secondly, in addition to studies utilizing proteomics to explore roles of etiological factors, risk factors and important molecules in the pathogenesis of NAFLD, studies of NAFLD pathogenesis by the means of subcellular proteomics, modification-specific proteomics, and the integration of proteomics and transcriptomics were also shown. Thirdly, methods for analyzing differentially expressed proteins and implications of these results were highlighted. It's hoped that this review can help to promote the application of proteomics in exploring disease pathogenesis and discovering novel biomarkers and treatment targets.

Abstract:The basic properties and mechanisms of human hearing are similar to the mammals, therefore, the hearing researches performed on and the results obtained from the animals are very helpful to elucidate mechanisms of auditory processing of human. It was discusses briefly the studies and finding on the sound signal recognition and processing in central auditory neurons. Recognition of sound signal and pattern plays an important role in sound signal perception and processing of auditory center. The auditory neurons, as the base of sound signal and pattern recognition, can generate different responses to different sound patterns, even to fine sound parameters change of the same sound pattern. However, mechanism underlying sound signal recognition of auditory neurons is still not completely clear up to now. On the other hand, sound signal is the carrier of sound information, and different information may be carried by different sound components or parameters of sound signal. The previous studies have demonstrated that central auditory neurons have abilities to encode and discriminate the sound information embedded in different sound signals. Therefore, they can generate responses to sound frequency, amplitude, and duration in changing and encode these sound parameters. These similar results obtained from animals of different species also implied that auditory centers of the animals have intercommunity and universality in recognition, analyzing, and processing of sound signal.

Abstract:As a main approach for disease related biomarkers discovery, quantitative research has become a hot topic in proteomics. With the development of experimental methods, the quantitative data processing methods are updated and improved constantly. Two categories of label-free quantitative algorithms are introduced at first, including database-free methods and database searching-based methods. Then, the analysis strategies and the details of the two kinds of methods are described, the advantages and disadvantages are also investigated, the frequently used tools and the corresponding internet resources are summarized. At last, some suggestions for improving the data processing of label-free quantitative proteomics are proposed.

Abstract:Avian influenza is a highly contagious disease of birds caused by A influenza viruses. The circulation in humans by the highly pathogenic H5N1 avian flu in the past few years have caused most pandemics and have heightened fear that the next influenza pandemic is due. Antibodies could be used as an efficient anti-virus agent in clinical therapy. The full-length HA of the A/Jiangsu/1/2007(H5N1) about 1.7 kb was amplified, subcloned to the pFastBac vector and recombinant bacmid DNA was selected. The recombinant HA was expressed and purified HA about 70 ku was used as the antigen to immunize Balb/c mice. The whole H5N1 virus was used to select 5 mono-antibodies (mAbs)，and all of them were tested using microneutralization assays. 8G10D7, one of the antibodies, had broad neutralizing effect against clade 2 and clade 9 H5N1 avian influenza A viruses, and the IC₅₀ was from 1∶256 to 1∶64. When detected with 8G10D7, all 4 viruses showed 70 ku and 43 ku protein band, which confirms that the binding site of the scFv antibodies were located at the HA1 domain. The nucleuses of MDCK cells infected by 4 viruses were colored purple, and red around the nucleus. 8G10D7 showed HI activity to the 4 viruses, the HI has a positive correlation with neutralization concentration IC₅₀, which also further confirms that the binding site of the scFv antibodies were located at the HA1 domain. When the mAb 8G10D7 was used for the study of prophylaxis and therapeutic effect on influenza A viruses infection in an embryonated chicken eggs model. It had a complete 100% protection effect on the H5N1 viruses in avian host in the prophylactic and therapeutic groups. The 100% preventive protection effect could be reached when challenged with H5N1 avian influenza A viruse in human host in the prophylactic groups, and there is also a 87.5% protection effect with H5N1 viruses in human host in the therapeutic groups. Thereby, the study suggests that the mAb 8G10D7 could be used in therapies to counter the H5N1 influenza A virus, and the epitope could be the key point for the design and implementation of vaccines.

Abstract:To determine whether TGF-βs and the effect on endothelial cell migration were altered during fresh frozen plasma (FFP) refrigeration, ELISA assay was carried out to quantify TGF-βs protein levels in FFP stored at 4℃ for up to 5 days. Human pulmonary microvascular endothelial cells (HPMECs) were treated with various concentrations of Day 0 and Day 5 FFPs or 10% Lactated Ringer's (LR) and were subjected to migration assay or Western blot analysis. ALK5 siRNA or ALK5 inhibitor were used to block FFP-induced Smad3 signaling in EC cells. It was found that TGF-β1 protein levels increased in a time-dependent fashion at a rate of 244.31 ng/(L·d) (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.

Abstract:Whether the immunogenicity of DNA vaccine can be further increased by novel deliver system- bacterial ghost is discussed. First, the bacterial ghost displaying ureB epitope by CS3 fimbriae was constructed, namely, plasmids pCSXureB harboring ureB epitope and plasmids pAcYclysis harboring lysis cassette were transformed into 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.

Abstract:SMAD-4 plays an important role in tumor suppression though controversies still exist regarding its behavior in carcinogenesis and its relationship with the phosphatase and tensin homolog deleted on chromosome 10 (PTEN), which is regarded as a key controller of cell cycle progression and cell growth. The role of SMAD-4 on PTEN expression was investigated through TGF-β signaling in 293T cells and in malignant gastric carcinoma MGC-803 cells. Results showed that SMAD-4 and TGF-β enhanced the expression of PTEN in 293T cells, while they suppressed PTEN expression in MGC-803 cells. However, this suppression was relieved upon the inhibition of RAS/ERK pathway. Moreover, the maximum expression of PTEN was achieved by the cooperation of SMAD-4 with TGF-β when SMAD-4 was translocated into the nucleus. Enhancement of early apoptosis of about three folds was achieved with this cooperation, compared with the action of TGF-β alone in MGC-803 cells. These findings shed light on the role of SMAD-4 as a co-Smad protein in TGF-β protein-signaling and in PTEN regulation.

Abstract:The role of CyclinB1 in conferring drug resistance in the treatment of liver cancer was investigated. siRNA delivery was used to knockdown the expression of CyclinB1, and flow cytometry analysis was employed to assess cell apoptosis and cell cycle distribution .Colony formation assay and cytotoxicity assay were used to determine cell growth ability. It was found that siRNA induced CyclinB1 down regulation triggered cell arrest at G2/M by 40%～50% and greatly inhibited cell colony growth ability. Daunorubicin in combination with CyclinB1 siRNA induced more apoptosis than that treated with Daunorubicin alone, whereas this combinational effect decreased in HL-7702 cells, a normal human liver cancer cell line. Those data support the notion that targeting CyclinB1 down regulation combined with chemotherapeutical agents would be a promising new strategy in the treatment of liver cancer.

Abstract:Eukaryotic genome DNA is packaged into condensed chromatin, which creates a natural barrier for functional factors to access to DNA during replication, transcription, repair and recombination. Interactions of these factors with DNA require relaxation of local chromatin structure, and this type of chromatin dynamic alteration is called chromatin remodeling. Increasing evidences have indicated that chromatin remodeling plays key role in DNA damage repair by facilitating the recruitment of DNA damage response proteins to DNA lesions. To investigate the association between chromatin remodeling and DNA repair and its coupling mechanisms, a Lac-repressor and Lac-operator based system was employed. Through this system, the Lac repressor-target gene- EGFP fusion protein can bind to the Lac-operator elements integrated in the genome of AO3_1 cells, and potentially reflects the process of chromatin remodeling after DNA damage. Using this system, it was found that TIP60 strongly promotes the large scale chromatin relaxation, and the p53 inducible gene 3 protein (PIG3) can also promote the large scale chromatin relaxation in the process of cellular response to DNA damage induced by 10 Gy γ-ray irradiation. Taken together, an efficient method was established to screen potential chromatin remodeling proteins associating with DNA damage repair.

Abstract:The method to obtain tumor cells from the primary culture of xenograft is commonly used. As primary culture is distinct from usual culture of single-layer cells, it also follows different patterns of cell growth. In order to observe the cell growth law of xenograft, a cell line (293-BAC) which stably expressed GFP was firstly used to formed transplantation tumor in nude mice. Primary culture of the tumor tissue was then conducted. Due to the stable GFP expression in the 293-BAC cells, the cell growth was observed through GFP readout and by analyzing the morphology of the cells from tumor tissue. Tumor cells in the masses of tissue were released to the out rim, gradually extending to the vicinity space. Cells in thin pieces which were scattered during planting were able to adhere to the flask wall and directly grew. There were about 1%～3% of mouse-origin cells growing among the tumor cells or gathering at the outskirt of newly-grown tumor cells. The mouse-origin cells disappeared after 5 passages of subculture. After the process of tumor forming and serial subpassages, the tumor cells were pure-sourced with stable growth property, thereby becoming good material for further research. Cell growth patterns of reference value for transplantation tumor or related research were provided.

Abstract:The c-Jun N-terminal kinase, JNK, is one of the mitogen-activated protein kinase superfamily members. JNK signaling pathway plays important roles in a variety of biological activities such as cell growth, differentiation and apoptosis. Post-translational modification by the small ubiquitin-related modifier (SUMO), termed as sumoylation, regulates multiple cellular and physiological processes. A very recent report in Development by Huang et al. establishes a link between sumoylation pathway and JNK pathway through the action of homeodomain-interacting protein kinase (Hipk), which advances our understanding of the JNK signaling regulation in Drosophila.

Abstract:This review focuses on the relationship between excess endogenous formaldehyde and memory decline, as well as multi-factors inducing formaldehyde abnormal accumulation and cognitive impairments. Elevation of endogenous formaldehyde concentrations in Alzheimer's animal models and clinical patients was discussed. Injection with formaldehyde (referred to the detected level in the AD animal models) into normal mice obviously induced spatial memory decline. Aging, and some of genetic factors, diet and environmental pollutants led to formaldehyde abnormal accumulation. Formaldehyde scavengers could rescue spatial memory of APP-transgenic mice. These studies addressed that excess formaldehyde-induced damage of brain is one of the critical factors of cognitive impairments of sporadic senile dementia. Scavenging excess formaldehyde may be a novel therapy for Alzheimer's disease.