Objective To study the effect of docosahexaenoic acid ( DHA ) and eicosapentaenoic acid (EPA) on cognition impairment and huntingtin associated protein 1 ( HAP1 ) expression in hippocampus of rat model with Alzheimer' s disease(AD). Methods Forty healthy Sprague-Dawley rats were randomly divided into control group, AD group, DHA group and EPA group on average. Alcl3 was injected intraperitoneally and D-galac tose was injected into subcutaneous to establish the model of rat with AD. ABC method of immunohistochemistry was used to observe the expression level of HAP1 in the hippocampus. Morris water maze was used to study the spatial learning and memory in rats. Results Compared with control group,the HAP1-positive neurons of hippo campus decreased and poorer performance in Morris water maze test was observed in AD group. Compared with AD group, DHA and EPA treatments significantly caused the decreases in escape latency and searching distance in the Morris water maze test. The number of HAP1- positive cells in DG region of DHA group(43.57 ±6.14) increased obviously compared with AD group( 28.56 ± 4.23 ) (P ＜ 0.05 ＝. Conclusion The immunoreactivities of HAP1 decrease in hippocampus of AD rat. The applications of DHA and EPA in AD rats significantly improve the ability of learning and memory;and the mechanisms may be related to the decrease of HAP1 expressions in hippocampus.

Objective:To clarify the differences of host immune responses at different stages of HBV infection. Methods:We constructed three HLA-A*0201/HBV tetramers with immunodominant epitopes of core18-27,polymerase 575-583 and envelope 335-343,and analyzed antigen specific CD8+ T cells and the expression of CD127 in peripheral blood mononuclear cells ( PBMCs) from patients infected with HBV using these HLA-A*0201/HBV tetramers. Results: The frequencies and expansion ability of antigen specific CD8+ T cells in most self-limited HBV infected individuals were higher than that in chronically HBV infected patients. In low copy period the frequencies of antigen specific CD8+ T cells were similar to those in immune clearance phase at a high viral load and liver damage and in immune clearance phase, which had no significant correlation with virus quantitation and ALT level. In chronic infection the ability of antigen specific CD8+ T cells proliferation was inversely proportional to the viral titer. In most self-limited HBV infected individuals the IFN-γsecretion functions of antigen specific CD8+ T cells were higher than in chronic infection,but in immune tolerance phase these cells lost the ability. HBsAg level was different at different stages after HBV infection:it was highest in immune tolerance phase,but in immune clearance phase,activity period and low copy period the correlation with HBV DNA replication gradually declined. The frequency of CD8+ CD127+ T cells in chronic HBV infection was lower than the control group and self-limited infection group,especially in immune tolerance with HBeAg+ and immune clearance phase. Conclusion: The frequencies of antigen specific CD8+ T cells are not the main determinant of immune-mediated protection in chronic HBV infection,memory antigen specific CD8+ T cells are not clear or missing,which provides the possibility for therapeutic vaccines and immunization therapy.

Objective To investigate the effects of extracts of Ginkgo biloba leaves ( EGb761 ) on learning,memory and the hippocampal long-term potentiation (LTP) in chronic alcoholic rats.Methods 72 SD rats were randomly divided into control group,chronic alcoholism model group,low-dose EGb761 (100 mg/kg) and high-dose EGb761 (200 mg/kg) groups on average.55％ alcohol was given ( 10 ml/kg) by gavages once every day for 6 weeks to create the model of chronic alcoholism.In low-dose and high-dose EGb761 groups,EGb761 was used for gastric infusion 30 minutes before model.The control group and the model group were infused with the same volume of distilled water.Morris water maze (MWM) was used to study the spatial learning and memory in rats.Electrophysiological methods were used to observe the changes of LTP through the comparison of field excitatory postsynaptic potential (fEPSP) slope in hippocampal CA3 region before and after high frequency stimulation (HFS).Results ( 1 ) The escape latency (EL) of model group ( (93.31 ± 14.21 ) s) was longer than that of the control group( (35.28 ±6.51 )s).The mean slope of fEPSP (relative to baseline) after HFS in model group was (96.91 ±7.65) ％,in control group was ( 177.22 ± 10.12) ％ ; Compared with control group,the ability of learning and memory as well as LTP obviously descended in model group.(2) Compared with model group,EGb761(100,200 mg/kg) could shorten the EL.The slope of fEPSP after HFS in either low-dose (( 126.16 ± 5.84)％ )or high-dose (141.31 ± 5.75)％ EGb761 groups was enhanced significantly compared with the model group (96.91 ± 7.65 ) ％,P =0.034,P =0.009 ).Conclusion EGb761 can significantly improve the alcohol-induced memory impairment by means of accelerating the recovery of the pathological synaptic plasticity.

Objective:To prepare and test tetrameric sH-2K~d-HBc complex for the further measurement of the specific CTL response.Methods:PE labled streptavidin with 4 biotinylated binding sites can bind to 4 biotinylated monomer to form the corresponding tetramer.Mice were immunized via different methods of genetic immunization by use of the construted pcDNA3-C plasmid to get the specific CTLs.Then our prepared tetramer was applied to stain the specific CTLs by the analysis of flow cytometry.Results:We applied our prepared tetramer to stain the cells from the experimental groups and control group.The results showed the tetramer was able to discriminate the frequencies of specific CTL induced by the three immunol methods(0.24%,0.26%,0.36% vs 0.07%,P≤0.05).This demonstrated that the prepared tetramer could bind its targets specifically and efficiently.The three immunol methods induced different levels of immune responses.Compared with the traditional muscle injection,gene gun induced weaker humoral immune response and stronger cellular immune response,and hydrodynamic injection induced the strongest humoral and cellular immune responses.Conclusion:Have successfully constructed the sH-2K~d-HBc tetramer.The techniques and methods can be used for preparation of tetramers of other types of MHCⅠ molecules.

This study investigated the expression profiles of IL-10 gene in three human hepatoma cell lines including Huh7, HepG2, and HepG2 transfected with a plasmid containing hepatitis B virus (HBV) named HepG2.2.15. RT-PCR analysis demonstrated that IL-10 message RNA was absent in HepG2 and Huh7 cells, whereas it was present in HepG2.2.15 cells, which was consistent with ELISA result. Furthermore, except for lamivudine other antiviral treatments did not significantly decrease the HBV DNA level in HepG2.2.15 cells, while they had different effects on the expression of IL-10 protein, although stimulation by LPS had no significant effect. In addition, except for poly(I:C), the other treatments decreased the expression of IL-10 protein to different degrees, but had no significant effects on the expression of NF-κB and MyD88. Meanwhile, all treatments we used had effect on the expression of STAT1. In conclusion, IL-10 was expressed in HepG2.2.15 cells and STAT1 pathway might be involved in the regulation of IL-10 expression in HepG2.2.15 cells, but it was not the sole pathway, the exact mechanism warrants further study.

Hepatitis B virus (HBV)-specific cytotoxic T lymphocytes (CTLs) are believed to play a major role in viral clearance and disease pathogenesis during HBV infection. To clarify the differences in host immune responses between self-limited and chronic HBV infections, we constructed three HLA-A*0201/HBV tetramers with immunodominant epitopes of core18-27, polymerase 575-583 and envelope 335-343, and analyzed the HBV-specific CTLs in peripheral blood mononuclear cells (PBMCs) from patients infected with HBV. The frequencies and expansion ability of HBV-specific CD8(+) T cells in most self-limited HBV infected individuals were higher than those in chronic HBV-infected patients. HBV-specific CD8(+) T cells could be induced by in vitro peptide stimulation from chronic patients with a low level of serum HBV-DNA but not from those with a high level of serum HBV-DNA. In chronic infection, no significant correlation was found either between the frequencies of HBV-specific CD8(+) T cells and the viral load, or between the frequencies and the levels of alanine transaminase. Our results suggested that the frequencies of HBV-specific CTLs are not the main determinant of immune-mediated protection in chronic HBV infection and immunotherapeutic approaches should be aimed at not only boosting a HBV-specific CD8(+) T response but also improving its function.

Objective:To study the changes of cardiopulmonary function in patients with rheumatoid arthritis,and to analyze the correlation between the changes ofcardiopulmonary function with oxidative stress and the peripheral blood lymphocyte attenuation factor.Methods:130 cases of patients with rheumatoid arthritis were studied as case group, and 50 cases of healthy persons were studied as normal control group.Detected the heart function parameters of two groups,which contained EF%,SV%,FS%,E A,E/A;lung function parameters of FVC,FEV1,MVV,PEF;B and T lymphocyte attenuation factor expression and activation level.Peripheral Cytokine(IL-17 and TNF-α,IL-4,IL-35) and oxidative stress index (ROS,MDA,SOD,TAOC) were detected by enzyme linked immu-nosorbent assay.Results:The indexes of cardiac function in the case group were significantly lower than that in the control group.103 cases had abnormal cardiac function index in the case group,which accounted for 79.23% of the case group,while the E/A had the highest abnormal rate.The case group had thickening of LADd, increasing of peak A, decreasing of EF, E peak and E /A, than the normal control group,the differences were statistically significant (P<0.05).Compared with normal control group,pulmonary function parameters were significantly lower in case group.88 cases of case group had abnormal pulmonary function,accounting for 67.69% of the case group.Among them,the abnormal rate of PEF was the highest.Pulmonary function indexes of case group was significantly lower than that of the control group, the difference was statistically significant ( P<0.05 ).Correlation analysis showed that the correlation coefficient of cardiac function indexes EF with CD24+cells and CD19+CD24+cells were respectively -0.353 and -0.457,which had negative correlation,with ROS the correlation coefficient was 0.459,which had positive correlation.The correlation coefficient of the cardiac function indexes in FS with CD24+cells,and CD19+CD24+cells was -0.395 and -0.421,which had negative correlation; the correlation coefficient of peak A and CD19+cell was 0.423,which had positive correlation;the correlation coefficient of E/A and BTLA was 0.393,which had obvious positive correlation.SV and MDA,SOD were positively correlated.The parameters of lung function with hs-CRP and ESR had significantly negative correlation.The correlation coefficient of lung function parameters FVC with BTLA and CD19+CD24+were 0.513 and 0.596,which had a significant positive correlation,with the correlation coefficient and CD24+BTLA+, TNF-αwere -0.451 and-0.351,which had significantly negative correlation.The correlation coefficients of FEV1 with CD24+CD19+, TAOC and IL-4 were 0.535,0.466 and 0.519,which showed a positive correlation,with CD24+BTLA+,MDA were -0.461 and -0.358,which had significantly negative correlation.The correlation coefficient of PEF with SOD,TAOC,IL-4,IL-35 were 0.547,0.482, 0.643 and 0.452,which had significantly positive correlation,with MDA,ROS,IL-17 were -0.451,-0.423 and -0.417,which had a significant negative correlation ( P<0.05 ).Conclusion: RA imbalance of oxidative stress and cell immune disorders which runs through the whole process in the heart and lung injury.Therefore,in clinical treatment,treatment of joint symptoms in RA patients needs restore the body′s redox homeostasis,in order to increase the level of BTLA,activate B cell and,T cell,thereby inhibiting immune and inflammatory response,reducing the heart and lung function impairment.

Previous studies have shown that expression of the interferon-sensitive gene (ISG)I5 protease UBP43 is increased in the liver biopsy specimens of patients who do not respond to interferon (IFN)-α therapy. We hypothesized that UBP43 might hinder the ability of IFN to inhibit HBV replication. In this study, we investigated whether vector-based siRNA promoted by Hi (psiUBP43) could enhance IFN inhibiting HBV replication in cell culture. UBP43 was specifically silenced using shRNA. In HepG2.2.15 cells, the HBeAg and HBV DNA levels were significantly reduced by IFN after transfection of shRNA, imphicated that vector-based siRNA promoted by HI (psiUBP43) could enhance IFN inhibiting HBV replication in cell culture. These data suggest that UBP43 modulates the anti-HBV type I IFN response, and is a possible therapeutic target for the treatment of HBV infection.

The pathogenesis of HBsAg (+)/HBsAb (+) double positive hepatitis B virus infection was investigated by simulating HBsAg/HBsAb coexistence in vitro and establishing HBsAg/HBsAb double positive model in vivo. Eukaryotic expression plasmids PCI-SY, PCI-adw, PCI-adr, PCI-ayw, which expressed S gene product of different serotypes, were constructed and transfected into HepG2 cells. Recombinant proteins were purified from the transfected cells. At the same time, HBsAg mouse antiserum was obtained by immunizing mice with PCI-SY plasmid. HBsAg/HBsAb coexistence was simulated using these antigens and antiserum. Furthermore, the expression plasmids expressing different serotypes of S gene product including PCI-adw, PCI-adr, and PCI-ayw were injected into mice via tail vein. HBsAg and HBsAb in mice sera were tested at the first and 7th day respectively after antigen plasmids injection. Both in vitro simulation and in vivo animal models demonstrated that HBsAg antigen and HBsAb of the same serotypes could not coexist, but HBsAg antigen and HBsAb of different serotype could coexist. HBsAg/HBsAb double positive hepatitis B virus infection could be due to infection of viruses of different serotypes.

The recombinant plasmid pCI-TSLC1 carrying TSLC1 gene was stably transfected into human hepatocellular carcinoma cell line HepG2. Cell proliferation was analyzed by MTT assay. The ability of migration was determined by transwell and FACSort flow cytometry was used to detect the cell cycle distribution and apoptosis. Western blotting revealed that H4 expressed higher amounts of TSLC1 protein than H15 and H0 did. The growth of TSLC1-transfected cells was significantly suppressed in vitro, and the ability of migration was reduced as well. The re-expression of TSLC1 could induce cell apoptosis. It was concluded that TSLC1 strongly inhibited the growth and ability of migration of HepG2 cell line in vitro and also induced apoptosis, suggesting that TSLC1 could reduce the tumorigenicity of human hepatocellular carcinoma cell line HepG2 in vitro, which provided a basis for further exploring the roles of TSLC1 in hepatocellular cellular carcinoma.
Apoptosis/*genetics ; Cell Adhesion Molecules/*genetics ; Cell Proliferation ; Hep G2 Cells ; Immunoglobulins/*genetics ; Neoplasm Invasiveness/genetics ; Transfection ; Tumor Suppressor Proteins/*genetics