Hepatitis: Wei L

Anonymous00371, McCaughan GW, Omata M, Amarapurkar D, Bowden S, Chow WC, Chutaputti A, Dore G, Gane E, Guan R, Hamid SS, Hardikar W, Hui CK, Jafri W, Jia JD, Lai MY, Wei L, Leung N, Piratvisuth T, Sarin S, Sollano J, Tateishi R. · Centenary Research Institute, AW Morrow Gastroenterology and Liver Centre, Royal Prince Alfred Hospital, University of Sydney, NSW 2006, Australia. · J Gastroenterol Hepatol. · Pubmed #17444847 No free full text.

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Wei L. · No affiliation provided · J Gastroenterol Hepatol. · Pubmed #19120855 No free full text.

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Wei L. · No affiliation provided · Zhonghua Gan Zang Bing Za Zhi. · Pubmed #17302020 No free full text.

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Guo F, Wei L. · Hepatology Institute, Peking University People's Hospital, Beijing 100044, China. · Zhonghua Gan Zang Bing Za Zhi. · Pubmed #15225448 No free full text.

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Hou J, Yin YK, Xu D, Tan D, Niu J, Zhou X, Wang Y, Zhu L, He Y, Ren H, Wan M, Chen C, Wu S, Chen Y, Xu J, Wang Q, Wei L, Chao G, Constance BF, Harb G, Brown NA, Jia J. · Nanfang Hospital, Guangzhou, China. · Hepatology. · Pubmed #18080339 No free full text.

Abstract: Chronic hepatitis B and its life-threatening sequelae are highly prevalent in China. There is a need for effective new therapies to suppress hepatitis B virus (HBV) replication and ameliorate liver disease. In this study, we compared the efficacy of telbivudine, a nucleoside analogue, with lamivudine in Chinese patients. In this phase III, double-blind, multicenter trial conducted in China, 332 patients with compensated hepatitis B e antigen (HBeAg)-positive or HBeAg-negative chronic hepatitis B were randomly assigned to treatment with 600 mg of telbivudine or 100 mg of lamivudine daily for 104 weeks. The primary efficacy endpoint was reduction in serum HBV DNA levels at week 52 of treatment. Secondary endpoints included clearance of HBV DNA to undetectable levels, HBeAg loss and seroconversion, therapeutic response, and alanine aminotransferase (ALT) normalization. Viral resistance and safety were assessed. At week 52, among 290 HBeAg-positive patients, mean reductions of serum HBV DNA were significantly greater in telbivudine recipients than lamivudine recipients (6.3 log(10) versus 5.5 log(10), P < 0.001), and HBV DNA was polymerase chain reaction-negative in significantly more telbivudine recipients than lamivudine recipients (67% versus 38%, P < 0.001). ALT normalization (87% versus 75%, P = 0.007), therapeutic response (85% versus 62%, P = 0.001), and HBeAg loss (31% versus 20%, P = 0.047) were also significantly more common in the telbivudine group. Treatment effects showed similar patterns in the smaller HBeAg-negative group (n = 42). Viral resistance in telbivudine recipients was approximately half that observed with lamivudine; however, this difference was not statistically significant. Clinical adverse events were similar in the two treatment groups. CONCLUSION: In Chinese patients with chronic hepatitis B, telbivudine treatment for 52 weeks provided greater antiviral and clinical efficacy than lamivudine, with less resistance.

Wei L. · People's Hospital, Hepatology Institute, Peking University, Beijing 100044, China. · Zhonghua Gan Zang Bing Za Zhi. · Pubmed #17125613 No free full text.

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Sun J, Gao Y, Chen HS, Wang SX, Li RB, Jiang D, Wei L, Wang Y. · Hepatology Institute, Peking University Health Science Center People's Hospital, Beijing 100044, China. · J Clin Virol. · Pubmed #15994118 No free full text.

Abstract: BACKGROUND AND OBJECTIVES: Host-virus interactions play a central role in determining the prognosis of hepatitis B virus infection. Multi-factors activated immune cells (MAICs) are autologous peripheral blood mononuclear cells activated with anti-CD3 monoclonal antibody, interleukin-2 and interferon-gamma. The present pilot clinical trial was designed to determine whether adoptively transferred MAICs inhibit the replication of hepatitis B virus and is safe for patients. STUDY DESIGN: Fourteen patients with chronic hepatitis B were enrolled in the study. A total of (1.5-4.0)x10(9) peripheral blood mononuclear cells were isolated from each patient and activated by anti-CD3 monoclonal antibody, interleukin-2 and interferon-gamma in vitro for 10 days to produce MAICs. Cell phenotypes and levels of cytokine secretion were determined during cell culture. Patients were followed up for 1 year after the transfusion of MAICs. RESULTS: After 10 days of culture, peripheral blood mononuclear cells were expanded to a mean fold of 4.68+/-1.78 and activated effectively, as demonstrated by CD25 expression and cytokine secretion. Cell activation peaked on day 4. All patients accepted the MAICs transfusion with some slight adverse events, and fulminant hepatitis and bilirubinemia were not observed. Significant HBV inhibition was observed in 8 out of 14 patients, in which 5 patients achieved complete response (defined as that serum HBV DNA levels below the detection limit, occurrence of HBeAg seroconversion and normalization of ALT were observed together after MAICs transfusion and kept for at least 6 months) and 3 patients achieved partial response (defined as that serum HBV DNA levels below the detection limit, occurrence of HBeAg disappearance and normalization of ALT were observed together in 6 months after MAICs transfusion, but kept for less than 6 months). CONCLUSION: These findings strongly suggest that MAICs transfusion, which effectively inhibits the replication of hepatitis B virus, is safe for patients.

Rao HY, Wei L, Li J, Zhang LF, Chen HY, Zhu LM, Liu F, Sun Y, Wang H. · Peking University People's Hospital, Peking University Hepatology Institute, Beijing, China. · Hepatogastroenterology. · Pubmed #19579592 No free full text.

Abstract: BACKGROUND/AIMS: We aimed to assess liver fibrosis in biopsies from patients with chronic hepatitis C and relationship to different responses to interferon-beta-la. METHODOLOGY: 21 patients with chronic hepatitis C were divided into two groups randomly and treated with recombinant human interferon-beta-la (IFN-B-1a) or IFN-beta-1a plus ribavirin (RBV) for 24 weeks, then followed up for another 24 weeks. 42 liver biopsies of 21 patients before and after treatment respectively were evaluated on conventional histological assessment. Then we studied 21 patients liver biopsies by immunohistochemical analysis of alpha-smooth muscle actin (alpha-SMA) and collagen type III. RESULTS: A significant improvement in HAI fibrosis staging was detected after therapy in all sustained viral responders (SVR) and non-responders (NR) patients. The percentages of cases with HAI scores and fibrosis staging decreased obviously were 100.0% and 71.4% in SVR patients and 50.0% and 42.9% in NR patients. The patients with combination therapy or normal ALT on 48w would more often receive the HAI and fibrosis staging decrease. The significantly lower alpha-SMA-positive HSCs and mean expression level of collagen type III were detected in the post-treatment biopsies. The HAI, alpha-SMA, collagen type III values were significantly correlated with the values of the semiquantitative indexes of fibrosis. CONCLUSIONS: IFN-beta-1a therapy is effective for patients with chronic hepatitis C on liver histology regardless of viral response. The alpha-SMA-positive HSCs and collagen type III expression are responsible for liver fibrosis.

Wang J, Jiang D, Zhang H, Lv S, Rao H, Fei R, Wei L. · Peking University Hepatology Institute, Peking University People's Hospital, Beijing, China. · Proteomics. · Pubmed #19242931 No free full text.

Abstract: Hepatitis B virus (HBV) infection is a worldwide health problem and may develop to liver fibrosis, cirrhosis, and even hepatocellular carcinoma. To investigate the global proteome responses of liver-derived cells to HBV infection and IFNalpha treatment, 2-DE and MS-based analysis were performed to compare the proteome changes between HBV stably transfected cell line HepG2.2.15 and its parental cell line HepG2, as well as HepG2.2.15 before and after IFNalpha treatment (5000 IU/mL for 72 h). Compared to HepG2, 12 of 18 down-regulated and 27 of 32 up-regulated proteins were identified in HepG2.2.15. After IFNalpha treatment, 6 of 7 down-regulated and 11 of 14 up-regulated proteins were identified. Differentially expressed proteins caused by HBV infection were involved with cytoskeletal matrix, heat shock stress, kinases/signal transduction, protease/proteasome components, etc. Prohibitin showed a dose-dependent up-regulation during IFNalpha treatment and might play a potent role in anti-HBV activities of IFNalpha by enhancing the crossbinding p53 expression to achieve the apoptosis of HBV infected liver cells. Down-regulation of interferon-stimulated gene 15 (ISG15) in HepG2.2.15 and recovery by IFNalpha suggested its relationship with IFNalpha's anti-HBV effect.

Mu JS, Wang HF, Wang JH, Pan XB, Zhang L, Wei L. · PLA Postgraduate Medical School, Beijing 100039, China. · Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi. · Pubmed #19031712 No free full text.

Abstract: OBJECTIVE: To construct a eukaryotic expression vector for expressing hepatitis B virus (HBV) recombinant HBsAg-EGFP fusion protein and obtain a stable transfected Chang Liver cell line. METHODS: The coding region of HBsAg gene of HBV was amplified by PCR and was digested by BamH I/EcoR I . This fragment was inserted into pEGFPN1 with T4 ligase and transformed E-coli TG1. The positive recombinant plasmid was selected, then the recombinant plasmid was transfected into Chang Liver cell by Lipofectamine 2000 cells containing stable transformants were selected by the ability of resistance to G418 and isolated with a limited dilution. The stable transfected cell line expressing high level HBsAg-EGFP fusion protein was obtained. RESULTS: The eukaryotic expression vector named pEGFPN1-HBsAg was successfully constructed and the stable transfected Chang Liver cell line expressing pEGFPN1-HBsAg fusion protein was obtained. CONCLUSION: The stable transfected Chang Liver cell line could express pEGFPN1-HBsAg fusion protein, could be used to screen the proteins differentially expressed in HBsAg expression Chang Liver cells, which brought some new clues for studying the potential molecular mechanism of HBsAg protein.

Pan XB, Han JC, Gao Y, Wei L. · Peking University People's Hospital, Peking University Hepatology Institute, Beijing 100044, China. · Zhonghua Yi Xue Za Zhi. · Pubmed #18756990 No free full text.

Abstract: OBJECTIVE: To investigate the effects of high level hepatitis B virus (HBV) replication on the hepatocytes. QSG-7701 cells. METHODS: Human hepatocytes of the line QSG-7701 were cultured and transfected with the plasmid pUC18-HBV1.2 or pUC18 containing 1.2 full length HBV DNA by the standard calcium phosphate precipitation method. Other QSG-7701 cells were transfected with the plasmid pUC18 as controls. Cell growth curves were drawn for 7 days after transfection. Four 4 days after transfection, HBV DNA in the culture medium was detected by using fluorescence quantitative real-time PCR. Cell apoptosis was detected by using terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling and electronic microscopy. Differential expressed genes were analyzed by using Oliga signal pathway micro-array. RESULTS: The curves of cell growth showed that the amount of control QSG-7701 cells increased by (8.3 +/- 1.2) times, significantly faster than the pUC18-HBV1. 2 transfected QSG-7701 cells that increased only by (1.1 +/- 0.2) times (P < 0.01). Four days after transfection, the HBsAg positive rate of the pUC18-HBV1.2 transfected cells was 35.4% +/- 6.7%, and the apoptotic rate was 15.2% +/- 4.3%. The HBV DNA level in the culture supernatant peaked 4 days adder transfection with the maximum value of (5.8 +/- 2.6) x 10(6) copies/ml. Genes related to cell growth and apoptosis, such as CASP3 (2.7981) ,CASP7 (2.2643), 3-Apr (3.5013), CDC2 (0.4380), MAPK6 (0.4447), and MAP3K2 (0.2785), were differentially expressed. CONCLUSION: High replicated HBV markedly inhibits the growth of hepatocytes and induces cell apoptosis.

Peng D, Han Y, Ding H, Wei L. · Hepatology Institute, Peking University, China. · J Gastroenterol Hepatol. · Pubmed #18707599 No free full text.

Abstract: BACKGROUND AND AIMS: Hepatic steatosis is commonly seen in chronic hepatitis C (CHC) patients. It has been reported to be associated with both metabolic factors and viral factors, and affects the severity of fibrosis in CHC. However, the relationship between hepatic steatosis and chronic hepatitis B (CHB) is unclear. The aims of this study were to investigate the frequency of hepatic steatosis in CHB patients, to identify the factors associated with its presence, and assess the relationship between the stage of steatosis and the severity of fibrosis. METHODS: Medical records of 153 adult patients with CHB who had undergone a liver biopsy within the past 4 years were included in the study. RESULTS: Body mass index (BMI) and age of CHB patients with steatosis was significantly higher than the patients without steatosis (P < 0.05), as determined by the univariate analysis. Steatosis was found to correlate with the BMI values and alanine aminotransferase (ALT) levels, and ALT levels were associated with hepatitis B virus (HBV)-DNA levels and histology activity index (HAI) scores, stages of fibrosis were associated with the HAI score and HBV-DNA, as determined by the multivariate analysis. In contrast, there was no significant association between advanced stages of fibrosis and steatosis. CONCLUSION: Our data indicate that hepatic steatosis is more frequently present in CHB patients than in the general population. We hypothesize that steatosis in CHB patients may be due to metabolic factors and the ability of HBV to indirectly facilitate the development of steatosis. In the present study, steatosis in CHB patients was not found to be associated with the severity of fibrosis.

Wu GY, Zheng XJ, Yin CC, Jiang D, Zhu L, Liu Y, Wei L, Wang Y, Chen HS. · Hepatology Institute, People's Hospital, Peking University, Beijing, China. · J Chemother. · Pubmed #18676226 No free full text.

Abstract: The authors investigate the effects and mechanisms of the anti-hepatitis B virus (HBV) agent Bay 41-4109. HepG2.2.15 cells were used to investigate the antiviral effects of Bay 41-4109 by using real-time polymerase chain reaction (PCR), western blotting, and immunofluorescence. The C terminally truncated core protein was expressed and purified. Changes in hepatitis B capsid formation were assayed by dynamic light scattering and transmission electronic microscopy. Bay 41-4109 was found to be a highly selective and potent inhibitor of hepatitis B virus replication in HepG2.2.15 cells. This compound was equally effective at inhibiting HBV DNA release and the cytoplasmic HBcAg level, with 50% inhibitory concentrations of 32.6 and 132 nM, respectively. HBV DNA and HBcAg were inhibited in a dose-dependent manner, indicating that the anti-HBV mechanisms are associated with and dependent on the rate of HBcAg inhibition. Our results indicate that Bay 41-4109 treatment disassembled the core capsids and separated them into monomers or dimers, the form in which they could be further degraded into peptides. The core protein assembled in a misdirected manner cannot function effectively. Our results suggest that, based on its particular activities, Bay 41-4109 is a promising anti-HBV candidate.

Zhang HH, Guo F, Fei R, Ma H, Cong X, Wei L, Chen HS. · Hepatology Institute, People's Hospital, Peking University, Beijing, China. · Zhonghua Yi Xue Za Zhi. · Pubmed #18649763 No free full text.

Abstract: OBJECTIVE: To evaluate the inhibition of CD4+ CD25+ regulatory T cells (Treg) in the chronic hepatitis B patients. METHODS: Peripheral blood samples were collected from 22 patients with chronic hepatitis B (CHB) and 18 healthy blood donors to isolate the peripheral blood mononuclear cells (PBMCs). Flow cytometry was used to analyze the proportion of CD4+ CD127(lo)CD25(hi-int) Tregs in the CD4+ T cells so as to calculate the proportion of CD4+ CD25+ Tregs in the CD4+ T cells. BrdU incorporation method was used to evaluate the immune inhibition of the CD4+ CD25+ Tregs. CD4+ CD25- cells were isolated by magnetic bead sorting technique. The CD4- T cells and CD4+ CD25- T cells ere mixed and stimulated by HBVcore 18-27 peptide. The PBMCs of the CHB patients with the Treg depleted and Treg not depleted underwent detection of HBVcore18-27 specific cytotoxic T lymphocytes (CTLs). The IFN-gamma secretion of the CTLs in the PBMCs of CHB patients with Treg depleted and Treg not depleted was detected by HLA-pentamer and enzyme-linked immunospot assay (Elispot). RESULTS: The proportion of CD4+ CD127(lo)CD25(hi-int) Treg in the CD4+ T cells used to reflect the percentage of CD4+CD25+ Tregs in the CD4+ T cells of the CHB patients was 4.3% +/- 2.4%, significantly higher than that of the healthy controls (2.1% +/- 1.3%, t = 3.74, P <0.01). There was no significant difference in the inhibition of CD4+ CD25- T cells by autogenous CD4+ CD25+ T cells between the CHB patients and healthy controls. The frequency of CTLs induced by HBV core 18-27 of the CHB patients with their CD4+ CD25+ cells in circulation depleted was 0.74% +/- 0.31%, significantly higher than that of the patients whose CD4+ CD25+ cells in circulation were not depleted (0.17% +/- 0.08%, t = 4.75, P <0.01). The frequency of IFN-gamma secreting spots of HBVcore18-27-specific CD8+ T cells of the CHB patients with their CD4+ CD25+ cells depleted was (112 +/- 33), significantly higher than that of the CHB patients whose CD4+ CD25+ cells in circulation were not depleted [(23 +/- 14), t =7.828, P<0.01)]. CONCLUSION: The proportion of CD4+ CD25+ Treg in CHB patients is increased compared to the healthy blood donor. The proliferative capacity of CD4+ CD25- T cells is inhibited by the presence of CD4+ CD25+ Treg dose-dependently, and the inhibition of CD4+ CD25+ Tregs in the CHB patients is similar to the inhibition of CD4+ CD25+ Tregs in healthy donors. The elimination of Treg cells followed by stimulation with HBVcore18-27 peptide significantly improves the antivirus CTL responses in CHB patients.

Ma H, Wei L, Guo F, Zhu S, Sun Y, Wang H. · Peking University Hepatology Institute, Peking University People's Hospital, Beijing, China. · J Gastroenterol Hepatol. · Pubmed #18624896 No free full text.

Abstract: AIM: To compare the clinical features of hepatitis B e antigen (HBeAg)-negative and HBeAg-positive cirrhosis, and to define the survival and prognostic indicators of Chinese HBeAg-negative hepatitis B virus (HBV)-related cirrhosis. METHODS: Two hundred and seventeen patients with chronic hepatitis B cirrhosis were studied. Survival was calculated using the Kaplan-Meier method. Proportional hazards Cox regression procedure was used to identify independent predictors of survival. The relationship between HBV-DNA viral load and prognosis was also investigated. RESULTS: The mean follow-up time was 35 months (3-47 months). HBeAg-negative liver cirrhosis patients comprised the greatest number of cirrhosis patients. Median alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels, median total leukocytes (WBC), hemoglobin (Hb), platelet (Plt) and HBV-DNA levels and the proportion of HBV-DNA > 10(5) copies/mL were lower in HBeAg-negative patients. There were fewer complications in patients treated with lamivudine than in other patients. Survival rates were significantly reduced in patients with HBeAg-negative cirrhosis (P = 0.0024). The baseline Child-Pugh scores and more than one decompensation during follow up were independent variables correlated with survival of HBeAg-negative liver cirrhosis patients (P = 0.006 and P = 0.001, respectively). The HBV-DNA viral load did not correlate with any complications or mortality rates of HBeAg-negative patients. CONCLUSIONS: The clinical features of HBeAg-negative and -positive liver cirrhosis differ. Survival was significantly reduced for Chinese patients with HBeAg-negative than -positive cirrhosis. Factors contributing to the prognosis were baseline Child-Pugh scores and the presence of more than one decompensation during follow up.

Rao HY, Li J, Zhang LF, Chen HY, Zhu LM, Wei L, Sun Y, Wang H. · Peking University People's Hospital, Peking University Hepatology Institute, Beijing 100044, China. · Zhonghua Yi Xue Za Zhi. · Pubmed #18353212 No free full text.

Abstract: OBJECTIVE: To access the effect of pegylated interferon (PEGIFN) beta-1a on the reduction of liver fibrosis in chronic hepatitis C (HVC). METHODS: Twenty-one patients with chronic HVC were divided into two groups randomly and treated with recombinant human PEGIHN-beta-1a (n = 13) or PEGIHN-beta-1a plus ribavirin (RBV) (n = 8) for 24 weeks, and then followed up for another 24 weeks. The clinical manifestations were observed and the clinical effects were evaluated. Biopsy was conducted before and after the treatment to analyze the histological activity index (HAI) and staging of fibrosis according the modified Knodell scoring system. Immunohistochemical analysis was used to examine the levels of alpha-smooth muscle actin (alpha-SMA), marker of activation of hepatic stellate cells (HSCs), and collagen type III in the HSCs. RESULTS: Sustained viral response (SVR) was achieved in 7 patients, and end-of-treatment virologic response (ETVR) was achieved in 9 patients. The HAI after treatment was 4.3 +/- 2.2, significantly lower than that before treatment (6.6 +/- 2.2, t = 4.77, P < 0.01). The fibrosis score after treatment was 1.1 +/- 1.1, significantly lower than that before treatment (1.7 +/- 1.2, t = 1.92, P < 0.05). The alpha-SMA score after treatment was 14 +/- 8, significantly lower than that before treatment (20 +/- 11, t = 2.15, P < 0.05). The integrated light density of collagen type III after treatment was 10 +/- 10, significantly lower than that before treatment (16 +/- 12, t = 1.83, P < 0.05). The improvement levels of fibrosis, alpha-SMA score, and integrated light density of collagen type III of the patients with SVR were all better than those of the patients without SVR; however, the differences were all not significant. The patients with combination therapy, female patients, and the patients with the HCV RNA < 2 x 10(6) copies/ml before treatment all showed higher levels in histology response. HAI, alpha-SMA level, and collagen type III values were all significantly correlated with the values of the semiquantitative indexes of fibrosis (all P < 0.01). CONCLUSION: Resisting hepatitis virus C and inhibiting and reversing the fibrotic progress, IFN-beta-1a therapy improves the liver histology of chronic HVC regardless of the viral response.

Pan XB, Han JC, Gao Y, Wei L. · Peking University People's Hospital; Peking University Hepatology Institute, Beijing 100044, China. · Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi. · Pubmed #18322586 No free full text.

Abstract: OBJECTIVE: The present aimed to observe the effect of phosphatase inhibitor cyclosporine A on the subcellular location and on expression of HBcAg in HepG2.2.15 cells. METHODS: Thirty micrograms/ml of cyclosporine A (CSA) was added into HepG2.2.15 cell culture system and on days 2 and 4 HBcAg and HBsAg were respectively stained with fluorescent immunocytochemistry and observed under confocal microscope. Cell apoptosis was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick end labelling method. RESULTS: HBcAg was mostly expressed in cytoplasm in the control HepG2.2.15 cells. After 2 days CSA administration of the expression of HBcAg and HBsAg in cytoplasm significantly decreased and the signals of HBcAg in nucleus increased , whereas the HBcAg was still mainly expressed in nucleus in about 1/4 of the cells. Cell apoptosis was observed in about 30% of the cells. CONCLUSION: CSA improves the nuclear entry of core protein. The increase of HBcAg in nucleus was likely to be related with it's phosphorylation and cell aging or apoptosis.

Pan XB, Han JC, Wei L, Peng DD, Gao Y. · Peking University People's Hospital, Peking University Hepatology Institute, Beijing 100044, China. · Zhonghua Gan Zang Bing Za Zhi. · Pubmed #18226340 No free full text.

Abstract: OBJECTIVES: The hepatitis B virus core protein has been found in nuclei, cytoplasm, or both of hepatocytes transfected with HBV DNA. It is still unclear whether intact core particles could pass through nuclear pores and what could be the mechanism regulating the subcellular localization of the core protein. This study on the distribution of core protein in hepatocytes and its translocation has a potential advantage to learn more about the HBV life cycle. METHODS: Dimethyl sulphoxide (DMSO, 2%), which effects hepatic differentiation, and/or 1 micro mol/L heteroaryldihydropyrimidine Bay41-4109, which interferes with the assembly of core particles, were added into HepG2.2.15 cell culture system for 4 days. The hepatitis B virus core antigen (HBcAg) and hepatitis B virus surface antigen (HBsAg) were stained with fluorescent immunocytochemistry and then observed under a confocal microscope. HBcAg in cytoplasm and nuclei were respectively extracted and analyzed using Western blot. HBV covalently closed circular DNA (cccDNA) was detected by using selective PCR method. RESULTS: The HBcAg was mostly expressed in the cytoplasm and weak signals of cccDNA were detected in the control HepG2.2.15 cells. After DMSO treatment, the expression of HBcAg in cytoplasm was increased about 2.5-fold; the expression of HBcAg and cccDNA in nuclei also increased. With the use of Bay41-4109, the signal of HBcAg in cytoplasm decreased 2/3, but it increased in the nuclei, and cccDNA decreased in the nuclei. When the HepG2.2.15 cells were treated both with DMSO and Bay41-4109, cord-liked distribution of HBsAg was observed in the cytoplasm. HBcAg in cytoplasm was decreased 1/2 but the HBcAg in the nuclei increased about 5-fold, whereas the cccDNA was almost negative. CONCLUSION: In HepG2.2.15 cells, the core protein is mainly assembled as a formation of core particles in the cytoplasm and they are blocked by the nuclear membrane. Bay41-4109 interferes with the assembly of core particles and the dissociated core proteins are able to enter the nuclei. DMSO promotes the nuclear entry of core protein/core particles and facilitates the formation of cccDNA.

Tian Y, Zhang HH, Wei L, Du SC, Chen HS, Fei R, Liu F. · Hepatology Institute, Peking University People's Hospital, Beijing, China. · Viral Immunol. · Pubmed #18158729 No free full text.

Abstract: Hepatitis C virus (HCV) nonstructural (NS) genes are relatively conserved and play critical roles in cellular immune responses against HCV. The aim of the study was to evaluate the immunogenicity of the different HCV NS genes through transduction of DCs and presentation to T cells. Monocyte-derived DCs from healthy donors were infected with the recombinant adenovirus (Ad) harboring HCV NS3 (AdNS3), NS4 (NS4A and NS4B; AdNS4), NS5 (NS5A and NS5B; AdNS5), NS3/NS4 (AdNS3/NS4), and NS4/NS5 (AdNS4/NS5) genes, and then used to stimulate autologous lymphocytes in vitro. Antigen-specific cellular immune responses were detected by interferon-gamma (IFN-gamma), interleukin 4 (IL-4), and Granzyme B (GrB) enzyme-linked immunospot assays (ELISPOT). DCs expressing different HCV NS genes all induced positive immune responses. Furthermore, DCs transfected with AdNS3/NS4 were superior to DCs infected with AdNS3 or AdNS4 in inducing HCV-specific immunity. The same results were obtained when we compared DCs infected with AdNS4/NS5 to AdNS4 or AdNS5. DCs transduced with NS3/NS4 or NS4/NS5 had similar ability to elicit specific immune responses to HCV.

Pan XB, Wei L, Han JC, Gao Y. · Peking University Hepatology Institute, Peking University People's Hospital, Beijing, PR China. · J Med Virol. · Pubmed #18041004 No free full text.

Abstract: Fluorescence quantitative real-time PCR (FQ-PCR) is a recently developed technique increasingly used for clinical diagnosis by detection of hepatitis B virus (HBV) DNA in serum. FQ-PCR is also used in scientific research for detection of HBV DNA in cell culture. Understanding potential FQ-PCR interference factors can improve the accuracy of HBV DNA quantification in cell culture medium. HBV positive serum was diluted with culture medium to produce three test groups with HBV DNA levels of 5 x 10(7) copies/ml (high), 5 x 10(5) copies/ml (medium), and 5 x 10(3) copies/ml (low). Chromosome DNA was extracted from HepG2 cells and then added to high, medium, and low group samples at final concentrations of 0, 12.5, 25, 50, and 100 microg/ml. The samples were quantified by FQ-PCR and data were evaluated using statistical software. No marked changes were seen in the quantitative curves for high level HBV DNA samples when the samples were supplemented with 0-100 microg/ml of chromosome DNA. Interference was observed in medium level samples when 50 and 100 microg/ml of chromosome DNA was added. Interference was also observed in low level HBV DNA samples when the concentration of added chromosome DNA was greater than 25 microg/ml. The interference was eliminated when samples were digested by DNase I prior to PCR detection. In Conclusions, the presence of cellular chromosome DNA can interfere with the detection of HBV DNA by FQ-PCR. Removal of cellular chromosome DNA from culture media prior to FQ-PCR is necessary for reliable HBV DNA quantitative detection.

Lu L, Wei L, Peng G, Mu Y, Wu K, Kang L, Yan X, Zhu Y, Wu J. · The State Key Laboratory of Virology, College of Life Sciences, Wuhan University, Wuhan 430072, PR China. · Virology. · Pubmed #17964630 No free full text.

Abstract: Hepatitis C virus (HCV) causes chronic hepatitis, which often results in the development of liver cirrhosis and hepatocellular carcinoma (HCC) worldwide. In this study, we demonstrated that the non-structural protein NS3 of HCV enhances cyclooxygenase-2 (COX-2) gene promoter activity, COX-2 mRNA expression, COX-2 protein production, and prostaglandin E2 (PGE2) release in HepG2 cells in a concentration-dependent fashion. We also showed that transcription factor NF-kappaB is required for the activation of COX-2 regulated by NS3. In addition, multiple signaling pathways are involved cooperatively in the expression of COX-2 activated by the viral protein in a calcium-independent manner, which requires signaling components including JNK, ERK, and PKD2. A thorough investigation of mechanism involved in the activation of COX-2 regulated by HCV would provide insights into our understanding the processes of liver inflammatory response and hepatocellular carcinoma development caused by the viral infection and also into the development of novel therapeutics against HCV infection.

Ma H, Guo F, Wei L, Sun Y, Wang H. · Hepatology Institute, Peking University People's Hospital, Beijing 100044, China. · Zhonghua Yi Xue Za Zhi. · Pubmed #17922993 No free full text.

Abstract: OBJECTIVE: Comparing clinical features and prognosis between HBeAg-negative and HBeAg-positive cirrhosis in patients with chronic type B hepatitis. METHODS: 217 chronic type B hepatitis with cirrhosis patients were prospectively studied for 35 months (3 - 47 months). Comparing clinical features and prognosis between HBeAg-negative and HBeAg-positive cirrhosis in patients with chronic type B hepatitis. RESULTS: The numbers of HBeAg-negative cirrhosis in patients with chronic type B hepatitis were more than HBeAg-positive cirrhosis; The median ALT and AST levels of HBeAg-negative patients were lower than HBeAg-positive patients; The median WBC, HGB and PLT levels of HBeAg-negative patients were lower than HBeAg-positive patients; HBV DNA positive rate and proportion of HBV DNA > 10(5) copies/ml of HBeAg-negative patients was lower than HBeAg-positive patients; The mortality rate of HBeAg-negative patients was higher than HBeAg-positive patients; among HBeAg-negative patients group, the presence rate of ascite, portal hypertensive gastrointestinal bleeding and HCC of patients treated with lamivudine were lower than the other patients, the proportion of non-presence of complications patients treated with lamivudine were higher than the other patients, the proportion of presence of one-two complications patients treated with lamivudine were lower than the other patients; among HBeAg-positive patients group, the presence rate of ascite of patients treated with lamivudine were lower than the other patients, the proportion of non-presence of complications patients treated with lamivudine were higher than the other patients. CONCLUSION: Among the liver cirrhosis patients, the numbers of HBeAg-negative cirrhosis were more than HBeAg-positive cirrhosis; HBeAg-negative patients with cirrhosis have lower liver inflammation activity; HBeAg-negative patients with cirrhosis have lower WBC, HGB and PLT levels; HBV DNA positive rate and proportion of HBV DNA > 10(5) copies/ml of HBeAg-negative patients was lower than HBeAg-positive patients; The mortality rate of HBeAg-negative patients was higher than HBeAg-positive patients; Lamivudine treatment is beneficial in HBeAg-negative and HBeAg-positive cirrhosis patients.

Lian M, Zhou X, Wei L, Qiu S, Zhou T, Li L, Gu X, Luo M, Zheng X. · National Laboratory of Protein Engineering and Plant Genetic Engineering, Peking University, Beijing 100871, China. · Virol J. · Pubmed #17892580 links to  free full text

Abstract: BACKGROUND: Hepatitis B virus (HBV) infection is a serious health problem worldwide. Treatment recommendation and response are mainly indicated by viral load, e antigen (HBeAg) seroconversion, and ALT levels. The S antigen (HBsAg) seroconversion is much less frequent. Since HBeAg can be negative in the presence of high viral replication, preS antigen (HBpreSAg) might be a useful indicator in management of chronic HBV infection. RESULTS: A new assay of double antibody sandwich ELISA was established to detect preS antigens. Sera of 104 HBeAg-negative and 50 HBeAg-positive chronic hepatitis B patients have been studied and 23 HBeAg-positive patients were enrolled in a treatment follow-up study. 70% of the HBeAg-positive patients and 47% of the HBeAg-negative patients showed HBpreSAg positive. Particularly, in the HBeAg-negative patients, 30 out of 47 HBpreSAg positive patients showed no evidence of viral replication based on HBV DNA copies. A comparison with HBV DNA copies demonstrated that the overall accuracy of the HBpreSAg test could reach 72% for active HBV replication. HBpreSAg changes were well correlated with changes of HBsAg, HBV DNA and ALT levels during the course of IFN-alpha treatment and follow-up. HBeAg positive patients responded well to treatment when reduction of HBpreSAg levels was more pronounced. CONCLUSION: Our results suggested that HBpreSAg could be detected effectively, and well correlated with HBsAg and HBV DNA copies. The reduction of HBpreSAg levels in conjunction with the HBV DNA copies appears to be an improved predictor of treatment outcome.

Liu LJ, Zhang R, Li JQ, DU SC, Jin D, Wei L. · Hepatology Institute, People's Hospital, Peking University, Beijing 100044, China. · Zhonghua Yi Xue Za Zhi. · Pubmed #17785063 No free full text.

Abstract: OBJECTIVE: To investigate the distribution of Hepatitis C Virus (HCV)genotype 1a in Yanbian area, Jilin province. METHODS: The genotypes of the serum specimens of 47 patients with HCV from Yanbian area were determined by Scheme ABC of RFLP based on HCV 5' noncoding region (5'NCR). The samples of type 1a (Y2, Y4, Y6, and Y8) were amplified from the 5'NCR and NS5B regions by RT-PCR, and then sequenced directly. These sequences were subjected to phylogenetic analysis with 27 reference sequences of HCV complete genomes from GenBank. RESULTS: 44 specimens were HCV RNA positive, among which 19 specimens (43.2%) were of the type 1a/1b, 12 (27.3%) were of the type 1b, 8 (18.2%) were of the type 2a/1b, 4 (9.1%) were of the type 1a, and 1 (2.3%) was of the type 2a, however, the genotypes 2b and 3 approximately 6 lacked. The phylogenetic analysis for 1a samples showed that according to 5'NCR they belonged to type 1a, while on NS5B belonging to type 1b. The most nucleotide identity of 5'NCR was respectively 0.990, 0.990, 0.990, and 0.990 between Y2, Y4, Y6, Y8 and the isolate HC-J1 (genotype 1a), while that of NS5B was respectively 0.936, 0.957, 0.936, and 0.936 between Y2, Y4, Y6, Y8 and the isolate HC-J4 (genotype 1b). CONCLUSION: In Yanbian area the distribution of HCV genotype is different from those in other areas and 1a/1b has even replaced 1b as the most HCV genotype here. The results of genotyping 1a on 5'NCR and NS5B are completely inconsistent. This phenomenon may be the consequence of recombination in evolution.

DU SC, Zhang R, Li JQ, Wei L. · Hepatology Institute, Peking University People's Hospital, Beijing 100044, China. · Beijing Da Xue Xue Bao. · Pubmed #17657275 No free full text.

Abstract: OBJECTIVE: To develop a hepatitis C virus(HCV) reverse transcription-polymerase chain reaction (RT-PCR) assay using Uracil-DNA glycosylase (UDG) for amplicon contamination control and evaluate the temperature and UDG concentrations for anti-contamination. METHODS: In this new HCV RT-PCR assay, reverse transcription, UDG anti-contamination and the first PCR were carried out at the same time. The layer candles were used to prevent the contamination in the second PCR. dU-DNA was used as quality control for UDG anti-contamination and templates to determine the sensitivity of the new HCV RT-PCR assay. HCV cDNA was detected by DNA enzyme immunoassay (DNA-EIA). RESULTS: Complete degradation of amplicon DNA was observed on the conditions of 0.2au UDG per reaction volume respectively at 37 degrees C and 42 degrees C for 40 min. The anti-contamination condition also could eliminate all detectible dU-DNA, including the highest concentration of amplicon DNA.The 1:10(4) dilution of the HCV RNA sample containing 2.110x 10(5)copies /mL copies of RNA could be detected. CONCLUSION: Our results indicate that this new RT-PCR assay can control the contamination stringently and is sensitive as well.