The ultrastructure of dissociated brain nerve cells from embryonic chicken cult ivated in vitro was studied both under SEM and TEM. Primitive synaptic junctions among the bipolar and multipolar neuronal cell bodies and its processes forming networks of fibers appeared after 4 days of culture. The presynaptic bags and desmosome-like cell junctions were observed simutaneously. Axo-dendritic, axo-somatic and axo-axonal synapses increased in number after 7 days of incubation onwards. Neuronal degeneration and proliferation of fibrous glial cells were evident in10 and 14 days cultures.

Objective To detect antibodies against the hypervariable region 1(HVR1) of hepatitis C virus in blood screening.Methods The HVR1 antibodies were detected by F4HVR1 antigen,and then were compared with other antibodies of HCV.Results Among HCV-RNA positive samples,HVR1 antibody was 96.8% positive.The positive rate of HVR1 antibodies in 90 suspected HCV-infected samples was 61.1%,which was close to those against C and NS3,and higher than NS4 and NS5(P

With light and electron microscope, we studied the morphology of the thymus of the neonatal mouse. The results showed: 1.the lobules of the thymus had not well developed and there was no distinct demarcation between the cortex and medulla; 2.a cyst composed of epithelial cells with microvilli or cilia might be frequently seen in the medulla; 3.occasionally small lymphocytes with some glycogen particles in their cytoplasm were observed; 4.only a few small-sized thymic corpusles existed in the medulla, The article also described the ultrastructure of the lymphocytes, epithelial reticular cells, macrophage, interdigitating cell and blood-thymus barrier in the thymus of the neonatal mouse.

Objective To study the clinicopathological and immunophenotypical features of gastrointestinal stromal tumor (GIST). Methods A SP Immunohistochemical method was selected to detect the 71 cases of GIST for a panel of antibodies CD117, CD34, Vim, Act, S-100. Results The patient's ages ranged from 21 years to 86 years (mean 55 years) including 34 male and 37 female. Most cases were diagnosed by bellyache and alimentary tract bleeding. Tumor size varied from 0.3 cm to 23 cm (mean 5.8 cm). GIST were composed of spindle cells and (or) epithelioid cells. Various sized eosinophilic globules were observed between the tumor cells and were designated as skeinoid fibers. The positive rate by immunohistochemical methods for CD117 and CD34 were 94.4 % and 73.2 % respectively. Conclusion GIST predominantly occurred in middle aged or older patients over 40 years. The histological characters are similar to smooth muscle tumor and schwannoma. CD117 and CD34 are more specific and sensitive markers. GIST may have been derived from interstitial cell of cajal or show differentiation toward interstitial cell of Cajal.

Objective To establish the enzyme-linked immunosorbent assay (ELISA) for detecting human serum fumarate hydratase (FH) antibody and evaluate its role in the diagnosis of autoantigen in autoimmune hepatitis (AIH).Methods The indirect ELISA was established using FH protein,and the reaction conditions were determined.Then,the anti-FH antibody were detected in the serum of 88 AIH patients,56 primary biliary cirrhosis (PBC) patients,50 chronic hepatitis B (HBV) patients,36 chronic hepatitis C (HCV) patients and 98 healthy controls(HC).The results were analyzed with chi-quare and Kruskal-Wallis H methods.Results The ELISA for detecting human anti-FH antibody was established successfully and the optimal reaction conditions were defined.The positive rate of anti-FH antibody in the AIH group (40％) was significantly higher than HC (3％,x2=38.44,P＜0.01),PBC group (7％,x2=18.45,P＜0.01),CHB group (2％,x2=23.59,P＜0.01) and CHC group (6％,x2=14.29,P＜0.01).Anti-FH antibody which was used to diagnose AIH revealed a sensitivity of 40％ and specificity of 94％.Conclusion We have established the ELISA,which is used to detect human anti-FH antibody.It can be detected predominantly in AIH,and this implies that anti-FH antibody may be useful in improving the diagnosis of AIH.

Objective To investigate immune responses and protective effect induced by two recombinant proteins of hepatitis C virus(HCV)in BALB/c mice.Methods BALB/c mice were immunized with recombinant proteins HCV-T and(or)HCV-F4HVR1 three times.Specific antibodies in sera were tested by enzyme-linked immunosorbent assay(ELISA).Five mice were sacrificed after 14 days of the last immunization.Splenic cells were isolated and levels of interferon(IFN)-γ,interleukin(IL)-4 and cytotoxic T lymphocyte(CTL)cytotoxicity assay were measured in vitro.The remaining mice were subcutaneously injected with 1.0×106 SP2/0-NS3 cells on the back to investigate the protective effects.The differences of means between groups were compared by LSD-t test.Results Compared with phosphate bufter saline(PBS)group,combined immunization with HCV-T and HCV-F4HVR1 induced higher levels of specific IgG against HCV-F4HVR1(t=3.815,3.762,P＜0.05),HCV-NS3-specific CTL response(t=3.971,P＜0.05)and IL-4(t=3.512,3.417,P＜0.05)and IFN-γ(t=3.813,3.426,3.671,P＜0.05)secretions.Conclusion High levels of specific humoral immunity and cellular immunity are induced in vivo after combined immunization with HCV-T and HCV-F4 HVR1,which could effectively prevent from the attack of SP2/0-NS3 cells.

Objective To study the hepatocyte cells infected by hepatitis C virus (HCV) positive serum. Methods Human hepatocyte 7701 was incubated with HCV RNA-positive and HCV antibody(Ab) negative sera BP52. Then, the expression of HCV antigen and the presence of HCV-RNA in cell and supernatant were assayed by RT-PCR, sequence analysis, immunofluorescent staining, Western blot, confocal laser microscopy. The ultrastructural changes of infected cells were observed by electro-microscopy. Results Plus-strand RNA and minus-strand RNA were intermittently detected in cell and/or supernatant on day 7-45 after infection. Sequence analysis demonstrated that the positive DNA nucleic acids were identified with HCV 5′-non-coding region(NCR) sequence. HCV core and NS3 protein were expressed in cytoplasm of infected cells. After 2 or 3 weeks, obvious intracellular ultrastructural changes and virus-like particles were observed. Conclusion human hepatocyte 7701 could support replication of HCV in vitro, which could be a useful tool for setting up cell model of HCV infection and studying the mechanism of HCV infection.

Objective To clone and express Mycobacterium tuberculosis fusion antigen ESAT-6 and CFP-10 ( EC) and to evaluate the biological activity of the fusion antigen EC in inducing specific cytokines secretion from THP -1 cells. Methods The fusion antigen EC gene was cloned into pET-30 a prokaryotic expression vector and expressed highly in E.coli BL21.Then, the THP-1 cells were stimulated with purified fusion antigen EC of different concentrations (10 and 20 μg/ml).Culture supernatants were collected after 12 h and 24 h, respectively.The secretion levels of IL-2, IL-4, IL-6, IL-8, IL-10, GM-CSF, TNF-αand IFN-γin THP-1 cell culture supernatants were detected using Bio-PlexProTM Assays kit.Results The M.tuberculosis fusion antigen EC was cloned and expressed successfully .The secretion levels of IL-6, IL-8 and TNF-αin EC infected THP-1 cells were significantly higher than those in THP-1 cells (P<0.05).The secretion levels of other cytokines did not change significantly .Conclusion The obtained M.tuberculosis fusion antigen EC has biological activity in inducing the THP-1 cells to secrete a higher level of IL-6,IL-8 and TNF-α.

ObjectiveTo discuss the relationship between the cross-reactivity of antibody against the hypervariable region 1 ( HVR1 ) of hepatitis C virus and early viral response ( EVR ) in patients undergoing antiviral therapy.MethodsBy ELISA and HCV HVR1 antibody cross-reactivity matrix reagent,the differences of anti-HCV hypervariable region antibody were tested in baseline serum from 46 patients with chronic hepatitis C before antiviral therapy.HCV genotyping and HCV RNA were analyzed by RT-PCR method.The HCV RNA assay was done at three time points:before treatment,pegylated interferon in combination with ribavirin therapy for 12 and 48 weeks.ResultsIn 46 cases of chronic hepatitis C patients,there were 16 cases with HCV 2a type,30 cases with l b and 33 patients obtained EVR.The EVR incidence of type 2a[ 93.8％ ( 15/16 ) ] was higher than that of type 1 b[ 60.0％ (18/30),x2 =4.316,P ＜ 0.05 ].In the EVR group of type 1b chronic hepatitis C patients,the positive number of average multi-target HVR1 antigen was ( 12 ± 4),which was significantly higher than that in the Non-EVR patients [ (7 ± 5 ),t =2.797,P ＜0.01 ].Bv the Fisher exact test,it was showed that patients'serum response to HVR1 antigens numbered 001,003,009,013,016 were higher in EVR group than those in non-EVR group,with statistically significant (P ＜ 0.05 ).ConclusionThe cross-reactivity of HVR1 antibody may play an important role in predicting the effectiveness of antiviral therapy.

Objective To establish a sandwich enzyme-linked immunosorbent (ELISA) method for early diagnosis of hepatitis C. Methods Immunization of Balb/c mice with hepatitis C core antigen were prepared by genetic engineering. Mouse monoclonal antibodies (McAb) to anti-HCV-core Ag were obtained. A sandwich ELISA kit for detecting HCV-core Ag was developed by using four strains of anti-HCV-core Ag McAb. One hundred and twenty five serum specimens with increased ALT but negative for anti-HCV, anti-HIV, and anti-Tp tests were tested. Results Nine of the 125 specimens were positive for HCV-core Ag. Conclusion The double sandwich ELISA kits for detecting HCV-core Ag may be useful for the early diagnosis of hepatitis C .