Changes of Toll-like receptor (TLR) 2 and TLR4 on the peripheral blood mononuclear cells in patients with chronic hepatitis B and chronic severe hepatitis B / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To study the changes of TLR2 and TLR4 on peripheral blood mononuclear cells (PBMCs) and their role in the pathogenesis of chronic hepatitis B and chronic severe hepatitis B.</p><p><b>METHODS</b>The expressions of TLR2 and TLR4 on 10000 CD14+ PBMCs were determined by flow cytometry in 30 healthy controls, in 31 patients with chronic hepatitis B and in 30 patients with chronic severe hepatitis B. The level of serum tumor necrosis factor alpha (TNF alpha) was determined by ELISA. The differences of expression of TLR2 and TLR4 on PBMCs and serum TNFalpha among the three groups of study subjects were determined by Student-t test. The correlations between TLR2, TLR4 and TNF alpha were determined by linear correlation test.</p><p><b>RESULTS</b>The values of mean fluorescence intensity (MFI) of TLR2 on PBMCs of the healthy controls, patients with chronic hepatitis B and patients with chronic severe hepatitis B groups were 21.5+/-2.7, 39.0+/-4.1, and 47.7+/-21.4; TLR4 of those groups was 2.3+/-1.1, 3.7+/-2.3, and 6.9+/-4.1. The serum TNF alpha(ng/L) of the respective groups was 53.8+/-38.1, 164.3+/-89.9, and 359.8+/-140.0. There was a gradual increase of these values from the group of healthy controls to the group of patients with chronic hepatitis B and patients with chronic severe hepatitis B. No significant positive correlations between TLR2, TLR4 and serum TNFalpha were found.</p><p><b>CONCLUSION</b>TLR2 and TLR4 may have a role in the pathogenesis of chronic hepatitis B and chronic severe hepatitis B.</p>

Changes of RANTES levels in livers of patients with chronic hepatitis B: the clinical significance and the possible mechanisms / 中华肝脏病杂志

<p><b>OBJECTIVES</b>To study the relationship between intra-hepatic levels of regulated on activation, normal T-cell expressed and secreted (RANTES) and the disease severity and liver inflammatory degrees in patients with chronic hepatitis B and the possible mechanism of the changes of intra-hepatic levels of RANTES.</p><p><b>METHODS</b>The expression of RANTES of the livers was studied using immunohistochemical stainings and morphometric quantitative measurements in liver specimens from 10 normal subjects and 64 patients with chronic hepatitis B with different degrees of liver inflammation and different clinical severity. The expressions of RANTES protein and mRNA in cell line HepG2, HepG2.2.15 and HepG2 treated with 10 ng/ml TNFa at different times were quantified by ELISA and one-step RT-PCR.</p><p><b>RESULTS</b>The expression of RANTES of the livers in patients was significantly higher than that in the normal controls. Hepatic RANTES levels increased significantly and the increases were parallel to the increases of the severity of the hepatitis, from mild, moderate to severe hepatitis (the positive units were 3.7+/-1.5, 15.6+/-6.9, 24.0+/-4.0, 37.9+/-11.1, respectively) and from G0 degree to G4 degrees of liver inflammation (the positive units were 3.7+/-1.5, 15.0+/-5.7, 21.6+/-5.9, 30.3+/-8.2, 40.9+/-12.3, respectively). The expressions of RANTES protein and mRNA of HepG2.2.15 were higher than that of HepG2. RANTES protein and mRNA were induced in HepG2 by TNFa.</p><p><b>CONCLUSION</b>RANTES may have an important role in the pathogenesis of chronic hepatitis B. The elevation of hepatic RANTES may be caused by hepatitis B virus and TNFa.</p>

Expression of anti-HBc single-chain variable fragment mediated by recombinant replication defective adenovirus in vitro / 中华肝脏病杂志

<p><b>OBJECTIVES</b>To define the expression of single-chain variable fragment (ScFv) against hepatitis B virus core protein (HBc) mediated by recombinant replication defective adenovirus carrying the anti-HBc ScFv gene in vitro and to define the activity of anti-HBc ScFv combining HBcAg.</p><p><b>METHODS</b>The recombinant adenoviruses carrying anti-HBc ScFv gene generated by homologous recombination in bacteria and packaged in 293 cells were transfected into HepG2 cells, and the anti-HBc ScFv was detected using SDS-PAGE and Western blot.</p><p><b>RESULTS</b>Green fluorescent protein (GFP) was observed in HepG2 cells after the transfection. SDS-PAGE displayed a protein strap about 2.7 x 10(4), and the result of Western blot displayed a positive reactive strap.</p><p><b>CONCLUSION</b>Anti-HBc ScFv can be expressed in cells mediated by recombinant replication defective adenovirus carrying the anti-HBc ScFv gene.</p>

Changes of antigen-specific cytotoxic T lymphocytes during acute flare-ups in chronic hepatitis B patients / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To detect HBV antigen specific cytotoxic T lymphocyte (CTL) changes in patients during acute flare-ups and to study their association with flare-ups and aggravations into grave hepatitis by quantitative analysis of HLA-A2* restricted HBcAg-specific CTL cells.</p><p><b>METHODS</b>The frequency of HBcAg-specific CTL cells in the peripheral blood mononuclear cells (PBMC) from 29 patients with persistent infection with HBV were quantified by flow cytometry using one HLA-A2*HBV peptide pentamers complex (Pro5TM MHC Pentamers).</p><p><b>RESULTS</b>There was a statistical difference of HBcAg specific CTLs between the patients with acute exacerbations (1.4%+/-0.8%) and the patients with immune tolerance (0.6%+/-0.4%) (t = 2.180, P = 0.01-0.05); There was no significant difference between the grave hepatitis group (1.3%+/-1.0%) and the chronic hepatitis group (1.4%+/-0.8%) regarding frequencies of antigen specific CTL (t = 0.215, P = 0.833-0.05). The level of antigen specific CTLs in PBMC in the 6 cases of chronic hepatitis B with acute exacerbations maintained a relatively high level (more than 0.7%) within the 12 week follow-up period.</p><p><b>CONCLUSION</b>HBcAg-specific CTLs may play an important role in hepatic flare-ups in patients with chronic HBV infection, but there was no direct relationship between antigen- specific CTLs and grave hepatitis.</p>

Clinical investigation of famciclovir in chronic hepatitis B patients irresponsive to alpha interferon treatment / 中华肝脏病杂志

<p><b>OBJECTIVES</b>To evaluate the efficacy and safety of famciclovir on the decreasing levels of serum HBV-DNA and ALT and HBeAg/antiHBe seroconversion in chronic hepatitis B patients irresponsive to 3 months treatment with alpha interferon.</p><p><b>METHODS</b>Two hundred and nineteen patients with chronic HBV infection, defined as positive HBsAg, HBeAg and HBV DNA, were enrolled and randomly half-and- half put into famciclovir and placebo groups. The two groups received either famciclovir 500 mg tid or a placebo treatment for 24 weeks, and then were followed-up for another 24 weeks with no treatment.</p><p><b>RESULTS</b>At the end of 24 weeks, the log value of HBV DNA dropped from 6.54+/-1.26 to 5.70+/-2.03 in the famciclovirt group and were elevated from 6.30+/-1.32 to 6.51+/-1.65 in the placebo group (P < 0.01). The rate of cases with persistence HBV DNA dropped 2 log of quantity in the famciclovir group and was 28.28% (28/99); it was 9.47% (9/95) in the placebo group (P < 0.01). Those with persistence negative HBV DNA was 28.28% (28/99) in the flamciclovir treated group and 14.74% (14/95) in the placebo group (P < 0.05). Those persistently being HBeAg negative were 7.69% (7/91) in the famciclovir treated group and 3.33% (3/90) in the placebo group (P > 0.05). The HBeAg/antiHBe seroconversion was 4.40% (4/91) in the famciclovir group and 2.22% (2/90) in the placebo group (P > 0.05). The percentage of cases with normal of ALT level was 15.15% in the famciclovir group and 6.35% in the placebo group (P < 0.05).</p><p><b>CONCLUSION</b>Famciclovir is effective in inhibiting HBV DNA replication and in decreasing serum ALT levels. The rate of HBeAg/antiHBe seroconversion in the famciclovir treated group was similar to that of the placebo group. Famciclovir was well tolerated without severe adverse effects during our treatment.</p>

Efficacy and safety in chronic hepatitis B adolescent patients with lamivudine therapy / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To analysis the efficacy and safety of lamivudine (made in China) therapy for 52 weeks in adolescent patients with chronic hepatitis B (CHB).</p><p><b>METHODS</b>One hundred and five teenage CHB patients were treated with lamivudine 100 mg once daily for 52 weeks. Patients with elevated ALT at baseline were in group 1 and those with normal ALT were in group 2. The changes of HBV DNA, HBV seromarkers and ALT at the end of 12, 24 and 52 weeks after lamivudine therapy were compared with those at baseline. Adverse events were recorded and evaluated.</p><p><b>RESULTS</b>At the end of 52 weeks of lamivudine therapy, HBV DNA-ve, HBeAg loss and anti-HBe seroconversion were observed in 92.0%, 24.4% and 22.0% in group 1 patients and 76.1%, 14.2% and 14.2% in group 2 patients respectively. No significant differences were found between two groups. At 12, 24 and 52 weeks, normalization rates of ALT were 59.0%, 66.7% and 76.0%, normal ALT with undetectable HBV DNA were 44.9%, 64.1% and 70.7% at the same time. During 52 weeks lamivudine treatment 26 mild adverse events were observed in 18 patients.</p><p><b>CONCLUSION</b>Lamivudine can inhibit HBV replication rapidly and normalize ALT in majority adolescent CHB patients. HBeAg loss or seroconversion of anti-HBe was observed in some of these patients. All patients in this study were safety and well tolerated.</p>

Chronic hepatitis B treated with domestic manufactured lamivudine in 2200 patients: a phase IV study / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To further verify the efficacy and safety of locally manufactured lamivudine on patients with chronic hepatitis B (CHB).</p><p><b>METHODS</b>2200 patients with CHB were recruited and received lamivudine orally 100 mg once daily for 12 months. The efficacy assessments included virologic response rate (defined by the absence of serum HBV DNA, HBeAg loss and HBeAg/HBeAb seroconversion), percentage of patients with normalization of alanine aminotransferase (ALT). Meanwhile improvement of quality of life (QOL) measured by mos SF-36 QOL questionnaire and liver histology evaluation were conducted in some patients. The safety assessments included adverse events, serious adverse events and laboratory abnormalities. All 2200 patients received at least one dose of medication and were all included in the safety population.</p><p><b>RESULTS</b>Ninety seven percent of patients (2137/2200) recruited were HBV DNA positive by dot blot (sensitivity GRT or equal to 1.0 pg/ml) at baseline. At the end of 12 months treatment, HBV DNA was undetectable in 80% patients (1538/1920) with HBV DNA positive before treatment. Among the 79%(1744/2200) of the patients recruited had positive HBV DNA accompanied abnormal ALT levels at baseline, 72% patients became ALT normal. And among the 84% (1843/2200) of the patients recruited were HBV DNA and HBeAg positive, anti-HBe negative, 16% (269/1650) patients achieved HBeAg/HBeAb seroconversion after 12 months of lamivudine treatment. The HBeAg/HBeAb seroconversion rate was positive correlation to the ALT level before treatment. A total of 304 patients completed the health-related QOL questionnaire. After 12 months treatment, lamivudine improved both their physical and mental health, especially for their mental health. 133 evaluable, paired liver biopsies were obtained for histological assessment, among whom 115 patients had abnormal ALT levels at baseline. Compared with pre-treatment most of their liver injury got alleviated (51.9%) or no further deterioration (36%), only 12% worsening. During the 12 months treatment, 9% patients withdrew from the study and 17% patients showed at least one adverse event, mild or moderate. There were no obvious difference between this study and the previously reported lamivudine Phase II or III study with regard to the kinds, incidence and severity of adverse events.</p><p><b>CONCLUSION</b>The efficacy and safety profile of the locally manufactured lamivudine 100 mg tablets are similar with those of the previously reported available lamivudine tablets imported in treating Chinese chronic hepatitis B patients.</p>

Reciprocal priming-boosting role of HBsAg and DNA vaccines / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To evoke more effective humoral and cell-mediated immunization against hepatitis B virus (HBV) infection.</p><p><b>METHODS</b>HBsAg-primed mice were boosted with HBs-DNA vaccine, and HBs-DNA-primed mice were boosted with HBsAg vaccine. Anti-HBs level was assayed by ELISA and cytotoxic T lymphocyte (CTL) response was tested by lactic acid dehydrogenase (LDH) releasing method two weeks after the boosted immunization.</p><p><b>RESULTS</b>Anti-HBs level and CTL responsive rate at the effector/target cell ratio of 100:1 were 0.38 and 36% in HBsAg/HBs-DNA vaccination group, 0.32 and 27% in HBs-DNA/HBsAg vaccination group, 0.48 and 1.5% in HBsAg/HBsAg vaccination group, 0.24 and 68% in HBs-DNA/HBs-DNA vaccination group, respectively.</p><p><b>CONCLUSION</b>Priming with HBs-DNA vaccine followed by boosting with conventional HBsAg vaccine results in greater antibody response (F = 21.19, P < 0.05), and CTL response after HBsAg vaccination can be improved by boosting with HBs-DNA vaccine (F = 165.59, P < 0.05). It brings to better efficacy by combining HBsAg vaccine with HBs-DNA vaccine.</p>

Therapy effect of lamivudine combination with alpha interferon on patients with chronic hepatitis B / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To evaluate the short-term therapeutic efficacy and safety of lamivudine (LAM) combining with alpha interferon (IFNalpha) on patients with chronic hepatitis B.</p><p><b>METHODS</b>90 chronic hepatitis B patients with HBV DNA and HBeAg positive were subdivided by 1:1:1 proportion into three groups: (1) LAM+IFN group: 6 months therapy of IFNalpha plus lamivudine followed by 6 months of lamivudine; (2) LAM group: lamivudine alone for 12 months; (3)IFN group: IFNalpha alone for 6 months.</p><p><b>RESULTS</b>At the end of treatment, the HBV DNA undetectable rate in LAM+IFN group (90.0%) was much higher than that in LAM group (80.0%) and IFN group (46.7%) (chi2 = 13.017, P < 0.001). ALT normalization occurred 90.0%, 80.0%, and 53.3% in LAM+IFN group, LAM group, and IFN group, respectively (chi2 = 9.932, P = 0.002). HBeAg/anti-HBe seroconversion rates achieved 46.7%, 13.3%, and 33.3% in LAM+IFN group, LAM group, and IFN group, respectively (chi2 = 7.937, P = 0.005). YMDD mutation was not detected in serum samples from LAM+IFN group patients.</p><p><b>CONCLUSIONS</b>LAM+IFN therapy for chronic hepatitis B is tolerated and more effective than IFN monotherapy in inhibiting viral replication and getting ALT normalization. The HBeAg/anti-HBe seroconversion rate with LAM+IFN therapy is higher than that with lamivudine monotherapy. LAM+IFN combination therapy seems to inhibit or postpone YMDD variants appearing in patients with chronic hepatitis B.</p>

Initiative factors of the damage sensitive stage of hepatocytes induced by interferon in patients with chronic hepatitis B / 中华肝脏病杂志

<p><b>OBJECTIVES</b>To probe into the initiative factors of the damage sensitive stage of hepatocytes induced by interferon in patients with chronic hepatitis B (CHB).</p><p><b>METHODS</b>Forty-four CHB patients with positive HBeAg and HBV DNA were treated with interferon. Serum ALT and viral markers levels of HBsAg, HBeAg, anti-HBc and HBV DNA were examined regularly. Liver biopsy was carried out just before the treatment.</p><p><b>RESULTS</b>The rate of HBeAg seroconversion was 75% at the sixth month, and 68.2% after one year of follow up. The rate of damage sensitive stage of hepatocytes was 47.7%. The average onset time was (3.14+-1.49) weeks after the treatment, and lasted for (8.24+-3.52) weeks. The ALT level raised (1.73+-1.13) times. The occurrence of damage sensitive stage of hepatocytes was indicator for good curative effect (Fisher exact probability, P=0.028). Damage sensitive stage of hepatocytes was more often developed in patients with moderate inflammation, overexpression of HBcAg in liver and higher level of HBeAg in blood stream before treatment. HBeAg and anti-HBc levels in peripheral blood decreased in the onset period of damage sensitive stage of hepatocytes.</p><p><b>CONCLUSIONS</b>The initiative factors of the damage sensitive stage of hepatocytes may be: HBeAg decreasing in peripheral blood induced by interferon may dismiss immune lutation of HBeAg and anti-HBc to cytotoxic T lymphocyte (CTL), which recognize HBcAg as target, thus activates the cytotoxicity of HBV-infected hepatocytes mediated by CTL.</p>