The significance of Toll-like receptor 4 (TLR4) expression in patients with chronic hepatitis B.

Purpose: To investigate the importance of Toll-like receptor 4 (TLR4) expression on hepatocytes obtained from Chronic Hepatitis B patients as well as on hepatocellular carcinoma HepG2 and HepG2.2.15 cell lines.

[Changes of RANTES levels in livers of patients with chronic hepatitis B: the clinical significance and the possible mechanisms].

Objectives: 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.

Methods: 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.

[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].

Objective: 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.

Methods: 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.

[Changes of antigen-specific cytotoxic T lymphocytes during acute flare-ups in chronic hepatitis B patients].

Objective: 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.

Methods: 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).

Results: There was a statistical difference of HBcAg specific CTLs between the patients with acute exacerbations (1.

[Expression of anti-HBc single-chain variable fragment mediated by recombinant replication defective adenovirus in vitro].

Objectives: 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.

Methods: 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.

Results: Green fluorescent protein (GFP) was observed in HepG2 cells after the transfection.

[Clinical investigation of famciclovir in chronic hepatitis B patients irresponsive to alpha interferon treatment].

Objectives: 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.

Methods: 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.

Risk factors for primary liver carcinoma in Chinese population.

Aim: To evaluate the risk factors for primary liver carcinoma (PLC) in Chinese population.

Methods: Chinese Biomedical Literature Database, China Hospital Knowledge Database and MEDLINE were searched. All the related literatures were screened, and the risk factors for PLC in Chinese population were studied.

Effects of c-myb antisense RNA on TGF-beta1 and beta1-I collagen expression in cultured hepatic stellate cells.

Aim: To investigate the effects of c-myb antisense RNA on cell proliferation and the expression of c-myb, TGF-beta1 and beta1-I collagen in cultured hepatic stellate cells (HSC) from rats.

Methods: Recombinant retroviral vector of c-myb antisense gene (pDOR-myb) was constructed, and then transfected into retroviral package cell line PA317 by means of DOTAP. The pseudoviruses produced from the resistant PA317 cells were selected with G418 to infect HSCs isolated from rat livers.

Construction of exogenous multiple epitopes of helper T lymphocytes and DNA immunization of its chimeric plasmid with HBV pre-S2/S gene.

Aim: To design and construct an exogenous multiple epitope of helper T lymphocytes (HTL), and to evaluate its effect on anti-HBs response through DNA immunization.

Methods: Artificial HTL epitope, PADRE and four other HTL epitopes from different proteins were linked together using splicing by overlap extension to generate exogenous multiple epitopes of HTL, MTE5. pcMTE5 and pcHB were generated by cloning MTE5 and fragments of HBV pre-S2/S gene into mammalian expression plasmid pcDNA3.

Promoting hepatic growth factor in the treatment of heavy type hepatitis and severe chronic hepatitis: a multicenter clinical study.

Background: The mortality rate of heavy type hepatitis is high. No special treatment is available except general treatment. This multicenter clinical study was designed to observe the safety and efficacy of promoting hepatic growth factor (PHGF) in the treatment of heavy type hepatitis and severe chronic hepatitis.

[Efficacy and safety in chronic hepatitis B adolescent patients with lamivudine therapy].

Objective: To analysis the efficacy and safety of lamivudine (made in China) therapy for 52 weeks in adolescent patients with chronic hepatitis B (CHB).

Methods: 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.

A 3-year clinical trial of lamivudine in treatment of patients with chronic hepatitis B.

Background: Lamivudine was approved for the treatment of chronic hepatitis B in China in 1999; however the long-term result has not yet been reported in detail. This clinical trial was to evaluate the long-term efficacy and safety of 3-year lamivudine treatment for chronic hepatitis B and the impact of emergence of YMDD mutation of hepatitis B virus (HBV).

Methods: This multi-center, randomized, double-blind, placebo controlled trial began from 1996 to 1999.

The levels of serum fibrosis marks and morphometric quantitative measurement of hepatic fibrosis.

Objective: To study the relationship between the serum levels of hyaluronic acid (HA), procollagen type III (PCIII), collagen type IV (CIV) and the histological degree of hepatic fibrosis evaluated by image analysis, and the clinical significance of serum HA, PCIII, CIV in the diagnosis of hepatic fibrosis in patients with chronic viral hepatitis.

Methods: The concentrations of serum HA, PCIII, CIV in 151 patients with chronic viral hepatitis were measured by radioimmunoassay. Liver biopsies were performed in all the patients.

Serum hyaluronic acid, procollagen type III and IV in histological diagnosis of liver fibrosis.

Objective: To assess the significance of serum hyaluronic acid (HA), procollagen type III (PCIII), collagen type IV (CIV) in the histological diagnosis of liver fibrosis.

Methods: The concentrations of serum HA, PCIII, CIV in 253 patients with chronic liver diseases were measured by radioimmunoassay. Liver biopsies were performed in all patients at the same time.

Liver fibrosis in chronic viral hepatitis: an ultrasonographic study.

Aim: To select valuable ultrasonographic predictors for the evaluation of hepatic inflammation and fibrosis degree in chronic hepatitis, and to study the value of ultrasonography in the evaluation of liver fibrosis and compensated liver cirrhosis in comparison with serology and histology.

Methods: Forty-four ultrasonographic variables were analyzed and screened using color Doppler ultrasound system in 225 patients with chronic viral hepatitis and compensated liver cirrhosis. The valuable ultrasonographic predictors were selected on the basis of a comparison with histopathological findings.

[CpG oligodeoxynucleotide enhances humoral and cellular immune responses against HBsAg in mice].

Objective: To induce stronger humoral and cell mediated immune response against HBsAg and seek more effective methods to treat hepatitis B virus infection.

Methods: 21-mer phosphorothioate oligodeoxynucleotide (ODN) and its control ODN were synthesized and used as an adjuvant of HBsAg and commercial hepatitis B vaccine to enhance their immune responses against HBsAg. Four groups of 33 mice received 2 doses of the mixtures (1.

[Initiative factors of the damage sensitive stage of hepatocytes induced by interferon in patients with chronic hepatitis B].

Objectives: To probe into the initiative factors of the damage sensitive stage of hepatocytes induced by interferon in patients with chronic hepatitis B (CHB).

Methods: 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.

[The long-term efficacy of lamivudine in chronic hepatitis B: interim analysis of 3-year's clinical course].

Objective: To evaluate the long-term efficacy and safety of 3-year lamivudine treatment for chronic hepatitis B and the impact of emergence of YMDD mutation of hepatitis B virus (HBV).

Methods: This multi-center, randomized, double-blind, placebo-controlled trial began in 1996. A total of 429 patients with serum HBsAg, HBeAg and HBV DNA positive were randomized to receive either lamivudine 100 mg daily (n = 322) or placebo (n = 107) in a 3:1 ratio for the first 12 weeks.

[Effect of exogenous epitopes of helper T lymphocyte on humoral immunity of HBV S gene DNA immunity].

Objective: To study the effect of exogenous epitope of helper T lymphocyte (HTL) on humoral immunity of HBV S gene DNA immunity.

Methods: Two universal HTL epitopes, amino acid residue (aa) 830-843 of the tetanus toxoid (TTE) and artificial epitope (PADRE), and 3 unique epitopes, aa1-20 of tubercle bacteria hot shock protein 65 (TBE), aa54-65 of rubella protein E2-4 (ME) and aa35-48 of trachoma hot shock protein 60 (CE) were chosen. Eukaryotic expression vectors were constructed by inserting single or multiple exogenous epitopes in HBV S gene just after the initial code of translation.

[Therapy effect of lamivudine combination with alpha interferon on patients with chronic hepatitis B].

Objective: To evaluate the short-term therapeutic efficacy and safety of lamivudine (LAM) combining with alpha interferon (IFNalpha) on patients with chronic hepatitis B.

Methods: 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.

Results: At the end of treatment, the HBV DNA undetectable rate in LAM+IFN group (90.

[Reciprocal priming-boosting role of HBsAg and DNA vaccines].

Objective: To evoke more effective humoral and cell-mediated immunization against hepatitis B virus (HBV) infection.

Methods: 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.

[Retroviral vector-mediated HBsAg expression and its stability].

Objective: To explore use of retroviral vector in gene therapy of hepatitis B.

Methods: The recombinant vector Plxsn-HBs was constructed by inserting HBV S gene into pLXSN. The pseudovirus, which was produced from PA317 after transferring with pLXSN-HBs by electroporation, were frozen at different temperature.

An extended two-year trial of lamivudine in Chinese patients with chronic hepatitis B.

Objective: To evaluate the long-term efficacy and safety of lamivudine therapy for the treatment of chronic hepatitis B and the clinical influence of emergence of tyrosine methionine aspartic acid (YMDD) motif mutation of hepatitis B virus (HBV).

Methods: This multicenter, double-blind, randomized, placebo controlled trial began in 1996. A total of 429 patients with HBsAg, HBeAg and HBV CNA positives were enrolled.