Objective Placental barrier is the nutrients and certain drugs,pathogens,hormones from the mother into the baby's only way. Isolated and cultured trophoblast cells in vitro to study its function and material exchange between maternal and fetal specific molecular mechanism of the cellular basis. The purpose of this study is to explore the in vitro isolation and culture of human placental trophoblast cells infected with hepatitis C virus （HCV） after HCV NS5 in trophoblast cell-specific expression. Methods Trophoblastic cells have been prepared from human term placentae digested with trypsin,and centrifuged on Percoll gradient. Trophoblastic cells were incubated in HCV positive serum,and then the HCV infection of syncytiotroblasts was detected with RT-PCR. Ultrastructure characteristics were observed with laser confocal microscopy. Results The HCV RNA was intermittently detected in the supernatent of the culture medium during the 40 days culture. The antibody of HCV NS5 could be observed around the nucleus with laser confocal microscopy. Conclusion Infected with HCV present in human trophoblast cells expressing HCV NS5. To further in-depth study of HCV molecular mechanism of mother to child transmission to lay an experimental basis.

Objective To establish HLA-A*2402 restricted HBV core antigen（HBcAg）-specific CTL clones. Methods PBMC obtained from a patient with chronic hepatitis B with HLA-A*2402 molecule positive were stimulated by using synthetic peptides（HBc117-125,EYLVSFGVW） and rHBcAg,and T cell clones subsequently were established by limiting dilution technique. T cell clones were then characterized by immunofluorescence staining,flow cytometry analysis and LDH release. Results After 3wk of in vitro stimulation,PBMC from the patient displayed peptide-specific cytolytic activity（43.7%,E/T ratio=60：1）;Eleven HBV-specific T cell clones were established from these activating PBMC by limiting dilution technique;Nine of these clones were CD8⁺T clones,and the other two were CD4⁺T clones;The nine CD8⁺T cell clones,including 2 Tc1,3 Tc2 and 4 Tc0,displayed peptide-specific cytolytic activity（33.9% to 90.2%,E/T ratio=2.5：1）. Conclusion HBc117-125 is capable of inducing Ag-specific cytotoxic T cells in PBMC of HLA-A*2402 positive patient with chronic hepatitis B. The Nine CD8⁺T cell clones display HBcAg epitope-specific cytolytic activity.

Objective To study the relationship between variation of calponin（CaP） expression and liver microcirculation disturbance of CHB. Method The liver biopsies samples from 200 patients with chronic hepatitis B（CHB） and 10 normal liver biopsies were stained for CaP expression by immunohistochemonical method. Results In control，CaP expression was showed in blood vessel and bile duct；In sinusoidal and microangiopathy mild （G1，G2） groups，CaP expression were showed dispersion or interrupt on hepatocyte，endotheliocyte and fat storing cell of domain of inflammation with vasculitis in hepatic tissue，and the positive was majority and a few showed negative and strong positive occupied 6%；But，in severe groups，not only CaP expression was showed diffuse by hepatocytes，but also there were all CaP expression in the liver sinusoidal walls and domain of capillarization and muscle fiber bundle. CaP expression ratio of strong positive was upgraded by degrees with sinusoidal expansion and vascular rebuilding or fibrosis. Between of the interclass was significantlly difference（P<0.01）. Conclusion The variation of CaP expression was related to abnormal microcirculation in livers of CHB.

Objective To determine the role of Pre-S1 protein in diagnosing viral replication,hepatic inflammation and fibrosis in patients with chronic hepatitis B. Methods 103 consecutive patients with chronic hepatitis B were included in the study,and liver biopsies were performed in all patients. Serial serum samples were studied with the quantitative determination of HBV DNA by a quantitative PCR assay,and determination of Pre-S1 protein by ELISA. The degrees of hepatic inflammation were divided into G1,G2,G3,G4,and degrees of hepatic fibrosis were divided into S1,S2,S3,S4. Results HBV DNA>10³copies/ml as positive criteria for diagnosing viral replication. The rate between Pre-S1 protein and HBV DNA detection was 88.1%;The rates between Pre-S1 protein and HBV DNA detection were 88.1%,96.3%,92.9%,83.3% in G1,G2,G3,G4 grading;The rates between Pre-S1 protein and HBV DNA detection were 84.2%,91.7%,90.9%,75.0% in S1,S2,S3,S4 staging. Conclusion The efficiency and sensitivity of Pre-S1 test were higher than of HBeAg,more sensitive than HBeAg test in diagnosing viral replication in patients with chronic hepatitis B.And Pre-S1 protein as a good sign in diagnosing degrees of of hepatic inflammation and fibrosis.

Objective To study the correlation between HBV PreS1 antigen,HBV M and HBV DNA. Methods PreS1 was detected by ELISA;HBV M by time-resolve dimmuno-fluorescent method;Liver fuction tests by kinetic method and HBV DNA by fluorescent quantitation PCR. Results The positive rates of HBV DNA and PreS1 in patients with HBeAg positive were 97.2% and 92.9%,respectively;That in patients with anti-HBe positive were 39.9% and 37.2%,respectively;and that in patients with HBsAg and anti-HBc positive were 62.7% and 54.2%,respectively. The positive rates of preS1 antigen in patients with chronic hepatitis B of mild,moderate and severe degree were not significantly different. Conclusion The detection of preS1 antigen can reflect the HBV replication and the implication of PreS1 is to be studied.

Objective To evaluate the diversity of biochemical virological indicator and the different clinical prognosis between HBeAg-positive and HBeAg-negative chronic hepatitis B group（CHB） patients with the long term therapy of lamivudine. Methods Sixty-three patients with chronic hepatitis B were treated with lamivudine,100mg/d and observed for three years. The serum HBV markers,HBV DNA,ALT,AFP and other biochemical indicators were detected. Results In the HBeAg-positive therapy group,the ratio of ALT normalization,the ratio of HBV DNA negative conversion,the incidence rate of hepatocellular carcinoma and the ratio of HBsAg negative conversion were 64.7%,64.7%,0%,0% after three years respectively;In the HBeAg-negative therapy group,these rates were 70.4%,77.8%,0%,0% respectively. There was no significant difference between the two groups. The ratio of YMDD mutation of HBeAg-positive patients was 43.3%,which was significantly higher than that（18.2%） in HBeAg-negative,x²=4.720,P<0.05. The proportion of patients with liver cirrhosis in HBeAg-negative group（37.0%） was higher than that in HBeAg-positive group（5.9%）,x²=3.866,P<0.05. Conclusion The patients with HBeAg-positive is more susceptible to YMDD mutation than HBeAg-negative patients;HBeAg-negative patients is more susceptible to liver cirrhosis than HBeAg-positive patients in the treatment of lamivudine.

Objective To explore the immunological location and expression of transforming growth factor TGF-β1 and platelet derived growth factor PDGF-BB in different stages on the hepatic fibrosis of chronic hepatitis B,and to discuss the possible effects and correlation in the development of liver fibrosis. Methods A total of 100 cases of chronic hepatitis B were obtained from the liver biopsy. The expression of TGF-β1 and PDGF-BB in fibrotic liver tissue were observed by immune histochemical,and the levels of liver function serum ALT,PTA were simultaneously measured. Results TGF-β1,PDGF-BB were positively related with the stages of hepatic fibrosis,correlation coefficient r for 0.824,0.856（P< 0.05）. There was obvious positive correlation between the expression of TGF-β1 and PDGF-BB,correlation coefficient r for 0.724（P< 0.05）. TGF-β1,PDGF-BB were negatively related with PTA,correlation coefficient r for -0.757,-0.656（P< 0.05）. TGF-β1,PDGF-BB were no related with ALT（P> 0.05）. Conclusion The expression of TGF-β1 and PDGF-BB may play important role in the process of hepatic fibrosis. TGF-β1 and PDGF-BB play a reciprocal promoting role in liver fibrosis.

Objective To explore main molecular mechanism of introhepatic microcirculation obstruction induced by chronic hepatitis B virus. Methods The liver biopsy tissues from patients with chronic hepatitis B,cirrhosis and healthy control were obtained. α-smooth muscle actin（α-SMA）,cyclooxygenase2（COX₂）,COX₁,and angiotensinII in the fibrotic livers and cirrhosis were examined by using immunohistochemistry. Results α-SMA,COX₂,COX₁ and angiotensin II in hepatic sinusoidal wall,little portal venules branch,hepatic arteries branch,central vein,vein wall beneath lobules were positive. They were obviously up-regulated as compared to control. Conclusions Intrahepatic microcirculation changes was found in chronic hepatitis B and cirrhosis.

Objective To analyze the relationship between liver MRI scores and Child-Pugh grades. Methods Liver MRI plain scans were performed in 48 patients with rival hepatitis-induced cirrhosis. Parameters of the length of liver,the thickness of ascites and the presence of collaterals were measured. The correlation were assessed between these parameters and Child-Pugh grades. Results The thickness of ascites and the number of collaterals were significantly and positively correlated with Child-Pugh grades（P<0.01）. The length of liver was significantly and negatively correlated with Child-Pugh grades（P<0.05）;Significant and positive correlation was found between liver MRI scores and Child-Pugh grades（r=0.935,P<0.01）;The accuracy of MRI scoring system in distinguishing between Child-Pugh grade A and grades B/C was 93.8%,with the sensitivity 95.2%,and the specificity 92.6%. Conclusion The clinical severity of liver cirrhosis can be well evaluated by using a liver MRI scoring system based on the presence of collaterals，the thickness of ascites and the liver length.

Objective To explore genotype distribution and clinical significance of HCV in Taiyuan region. Methods To amplify 5’ UTR of HCV RNA in 62 patients using RT-nested-PCR,to cut 5’UTR fragment into different length clips by using incision enzyme HaeⅢ and Bsh1236Ⅰ. Results In 62 patients with hepatitis C,43 cases were infected with genotype 1b,9 cases were infected with genotype 2a,5 cases was 1b/2a mixed-type,1 case was type 1a,1 case was type 3a,and 3 cases were not tested. The patients with 1b genotype were more serious than the patients with non-genotype 1b in the hepatic function and liver fatty degeneration. Conclusion In this locality hepatitis C were dominated by genotype 1b，and the pathogenetic condition of genotype 1b patients more serious than non-genotype 1b patients. HCV genotypes 1b accounted for 69.4% of HCV infections in patients from this geographical region. HCV genotype 1b was associated with more serious clinical performance.

Objective To observe the changes of the peripheral blood lymphocyte subsets in patients with chronic hepatitis C treated by pegylated interferon α-2a combined with ribavirin. Methods The peripheral blood lymphocyte subsets were detected in 27 patients with chronic hepatitis C and 26 blood donors. The patients received pegylated interferon α-2a and ribavirin combination therapy for 48 weeks. Results The CD4⁺T lymphocytes increased in patients with chronic hepatitis C as compared with control group（P<0.01）,CD8⁺T lymphocyte reduced（P<0.01）,and CD4⁺/CD8⁺ ratio increased（P<0.05）;after PEG-interferon α-2a combined with ribavirin therapy for 24 weeks,the CD4⁺ decreased（P<0.05）,and CD4⁺/CD8⁺ ratio increased（P<0.05）. Conclusion Pegylated interferon combined with ribavirin for chronic hepatitis C patients has a regulatory role in cellular immunity.

Objectives In order to know the changes of HBV DNA in patients with chronic hepatitis B. Methods To observe and compare the model for end-stage liver disease（MELD） scores and HBV DNA loads in 51 cases with acute-on-chronic liver failure and 56 with chronic hepatitis B of severe degree. Results MELD scores in peak and convalescent stages in patients with acute-on-chronic liver failure were 20.5±4.4 and 12.2±6.1,and HBV DNA loads were 6.2±1.5 log10 copies/ml and 4.5±1.5 log10 copies/ml,respectively;MELD scores in peak and convalescent stages in patients with chronic hepatitis B were 11.9±3.2 and 6.4±3.1,and HBV DNA loads were 6.5±1.5 log10 copies/ml and 4.9±1.8 log10 copies/ml,respectively（t=6.692~13.215,all P<0.000）. Conclusion No matter in patients with acute-on-chronic liver failure or chronic hepatitis B,their serum HBV DNA loads has spontaneously decreased in convalescent stages.

Objectives To explore the influence of HBV DNA on prognosis in early,middle and advanced stages of chronic severe hepatitis B patients. Methods To add up the information of HBV DNA in 197 patients in different stages of chronic severe hepatitis B from 2004 to 2008 in our hospital. To measure the three-month mortality of HBV DNA positive group and HBV DNA negative group in early stage,middle stage and advanced stage. To count the changes of model for end-stage liver（MELD） values on two weeks after admission. Results In early stage the three-month mortality of HBV DNA positive group and HBV DNA negative group were 34% and 7.4% respectively（P<0.01）. In middle stage and advanced stage the rates of two groups were 48.8% and 44.4%（P>0.05）,80% and 76.9%（P>0.05）. In early stage △MELD values on two weeks after admission of HBV DNA positive group and HBV DNA negative group were 3.6±4.8 and 0.8±4.1 respectively（P=0.01）. In middle stage and late stage the values of two groups were 1.5±4.2 and 1.1±6.5（P>0.05）,-1.6±7.3 and-1.1±5.8（P>0.05）. Conclusions HBV DNA positive influenced the prognosis of chronic severe hepatitis B in early stage,but not in middle stage and advanced stage.

Objective To explore the risk factor of nosocomial infection in patients with severe hepatitis. Methods 196 patients with severe hepatitis were analyzed retrospectively. Results The nosocomial infection rate in 196 patients was 50%（98/196）,out of which were spontanious bacterial peritonitis（88.8%）,biliary tract infection （42.9%）,intestinal infection（21.4%）,respiratory infection（12.2%） and others（9.2%）. The infection was related to the hospitalization,interventional treatment,hypoalbuminemia,administration of steroid and multiple infection. Conclusion Infection is important factor influencing the prognosis of patients with severe hepatitis.

Objective To evaluate the short-term efficacy of gamma knife radiosurgery in the treatment of patients with malignant obstructive jaundice. Methods 20 patients with malignant obstructive jaundice were treated by gamma knife radiosurgery. According to the size and location of the tumors,different diameters of collimator helmets were used. The radiosurgery plan was designed to cover whole target area at isodose curves of 50% to 70% with a margin dose of 3 to 5 Gy each time and the total margin dose of 35 to 50Gy,which was carried out at a frequency of 3 to 5 times each week. Results The serum levels of total bilirubin decreased from 215.4±143.5μmol/L to 98.4±54.5μmol/L after two week treatment（P<0.01）;CR was in 3 and PR in 14 patients with the effective rate of 85%;The pain intensity measured by numerial rating scale was 6.96±1.21 before the treatment,while it was 2.06±1.07 after the treatment. Conclusion Gamma knife radiosurgery is an effective and safe treatment in patients with malignant obstructive jaundice.