Objective:To develop a HCV combined HBV DNA-based therapeutic vaccine.Methods:The HBV core gene and HCV core cDNA were inserted into the eukaryotic expression vector with two multiple cloning sites mammalian expression vector under the CMV promoter and RSC promoter respectively, named pRSC-HBV/HCV. Cellular expression of pRSC-HBV/HCV was assessed by Western blot and immunofluorescent study following transfection into SP2/0 cells. The Balb/c mice were immunized by multiple sites intramuscular injection with pRSC-HBV/HCV and the immune responses were detected.Results:The 21 kD and 14 kD core protein were observed. Both anti-HBc Ab and anti-HCV core Ab were detected in all immunized mice.Conclusion:The investigation demonstrated that pRSC-HBV/HCV could express HBcAg and HCV core protein, humoral immune response could be detected in mice immunized with pRSC-HBV/HCV.

Objective:To study the immune responses of HBV based immunization co injecting CpG oligodeoxynucleotides in HBV transgenic mice.Methods:The HBV transgenic mice were immunized by multiple sites intramuscular injection with V HBs and CpG ODN or V HBs alone or V 1012 controls.The anti HBs Ab and HBsAg in serum from immunized mice was measured by ELISA.The expression of HBsAg in liver tissue was detected by immunohistochemistry method(SP).The histological activity index was calculated with Knodell's method.Results:The anti HBs Ab in serum could be detected and HBsAg in serum and liver tissues could not be measured in 2 out of 6 HBV transgenic mice immunized with V HBs+CpG ODN.There were not significantly changes in the level of anti HBs Ab and HBsAg in serum from HBV transgenic mice immunized with V HBs or V 1012 alone.The histological activity indexes in liver tissues from mice immunized with V HBs+CpG ODN were higher than that in mice inoculated with V HBs or V 1012 alone.The large number of lymphocytes could be found in liver tissues in V HBs+CpG ODN immunization group by microscopy.Conclusion:The immuno response,which seems to be responsible for the disappearance of HBsAg,might be elicited in HBV transgenic mice immunize with V HBs+CpG ODN.

Heat shock protein 70 (HSP70) is expressed differently in hepatocellular carcinoma (HCC) tissues. Since its expression and regulatory mechanism remain unclear, whether HSP70 can help with the early diagnosis, treatment, and prognostic evaluation of HCC has become a hot research topic. This article reviews the source of HSP70 and its family, abnormal expression of HSP70 in HCC, and its association with treatment methods and prognostic evaluation of HCC, in order to provide a reference for clinical diagnosis and treatment of HCC.

Objective To investigate the clinical value of glypican-3 (GPC3) in the diaganosis of hepatocellular carcinoma (HCC),the contents of GPC3 in the serum and tissues of HCC patients were detected.Methods ELISA and immunohistochemical staining were applied to detect GPC3 expressing level in the serum and tissues in 79 cases with HCC,35 cases with post-hepatitis cirrhosis and 30 normal liver specimens and the resuits were compared.The influential factor of GPC3 content in the patients with HCC was analyzed by logistic regression model.Results The serum level of GPC3 in patients with HCC was (143.02±40.26) μg/L which was signifcantly higher than that in patients with post-hepatitis cirrhosis [(6.15±4.31) μg/L] and healthy controls [(4.47±3.22) μg/L] (all P ＜ 0.01).The expression levels of GPC3 was signifeantly higher in post-hepatitis cirrhosis tumor-adjacent tissue and tumor-distant tissue.The expression levels of GPC3 was significantly positively correlated with tumor presence of distant mestasis (x2 =13.182,P ＜ 0.0) and clinical stage (x2 =4.250,P ＜ 0.05),and not correlated with sex,age,tumor size and the level of AFP inserum (P ＜ 0.01).Conclusion GPC3 is specific the diagnosis of HCC.The joint diagnosis of GPC3 and AFP will improve the sensitivity of HCC.Therefore,GPC3 could as a biomarker for evaluating HCC condition and prognsis.

BACKGROUNDTo explore the possibility of using retrovirus vector to carry HBV vector, and to prove that replication defective HBV could be normally packaged.

METHODSTwo kinds of full length of mutant HBV gene, which express dominant negative mutants, were inserted into retrovirus vector. After recombinant retroviruses were harvested, they were used to infect Hep G2 and 2.2.15 cell line. Then the expression of HBV core antigen in the Hep G2 cell was examined by immune fluorescence, and the existence of recombinant HB virion in the culture medium was examined by PCR.

RESULTSHigh titer of recombinant retroviruses were obtained in the culture medium of transfected PA317 cell line. Core antigen was detectable in the recombinant retrovirus infected Hep G2 cell. Recombinant HB virion was detectable in the culture medium of recombinant retrovirus infected 2.2.15 cell.

CONCLUSIONSThe results suggested that recombinant retrovirus could carry HBV vector and express HBV products. When structural protein is offered by wt-HBV, the recombinant retrovirus may function as HBV vector, therefore it could be used in anti?HBV gene therapy.

Genetic Therapy ; Genetic Vectors ; Hepatitis B Core Antigens ; biosynthesis ; Hepatitis B virus ; genetics ; Humans ; Recombination, Genetic ; Retroviridae ; genetics ; Tumor Cells, Cultured ; Virus Replication

BACKGROUNDTo study the significance of the expression of HCV core protein in PBMC of patients with chronic hepatitis C and to evaluate the relationship between HCV core protein expression and clinical states.

METHODSIdentification of HCV protein antigen (Ag) in PBMC of 66 hepatitis C patients by immunohistochemical method and clinical status of the patients with HCV protein positive expression were investigated. In 27 out off 66 patients the HCV RNA and HCV Ag in PBMC were detected with RT-PCR and immunohistochemical method.

RESULTSThe HCV Ag (core+NS3) was identified in PBMC in 51 out of 66 patients (77.27%). It was also demonstrated that HCV core protein in nucleus showed strong expression and NS3 protein in cytoplasm showed weak expression. The expression of core protein in nucleus of PBMC were 35.29% in advanced chronic hepatitis patients, which was significantly higher than that from moderate cases (5.88%).

CONCLUSIONSThis study suggested that the expression of PMBC-HCV core protein may be related to the clinical state of the patients. The nuclear expression of HCV core protein in PBMC of patients with hepatitis C may be related to the persistence and activity of the chronic hepatitis C virus and play an important role in the pathogenesis of cirrhrosis and hepatocellular carcinoma.

Hepatitis C Antigens ; blood ; Hepatitis C, Chronic ; virology ; Humans ; Immunohistochemistry ; Leukocytes, Mononuclear ; metabolism ; RNA, Viral ; blood ; Viral Core Proteins ; blood