Immune response pattern varies with the natural history of chronic hepatitis B.

Background: Chronic hepatitis B is a highly heterogeneous disease that can be divided into four phases: Immune tolerant (IT), immune active (IA), inactive carrier (IC) and hepatitis B envelope antigen (HBeAg)-negative hepatitis (ENEG).

Aim: To investigate the immune status of natural killer (NK) and T cells in different phases of chronic hepatitis B.

Methods: The frequency, phenotype and function of circulating NK cells, as well as nonantigen-specific and hepatitis B virus (HBV)-specific T cell responses were detected by flow cytometry in healthy and HBV-infected subjects.

Decreased peripheral natural killer cells activity in the immune activated stage of chronic hepatitis B.

Background & Aims: The natural course of chronic hepatitis B virus (HBV) infection is characterized by different immune responses, ranging from immune tolerant (IT) to immune activated (IA) stages. In our study, we investigated the natural killer (NK) cells activity in patients at different immunological stages of chronic HBV infection.

Methods: Blood samples obtained from 57 HBeAg positive patients with chronic hepatitis B (CHB), including 15 patients in the immune tolerant (IT) stage, 42 patients in the immune activated (IA) stage, and 18 healthy individuals (HI).

[The immunization effects of HBV core antigen and surface antigen fusion protein in mice].

Objective: To evaluate the immunization effects of HBV core antigen and surface antigen fusion protein.

Methods: The DNA fragments encoding HBsAg 100-162 aa; HBcAg 1-78 aa and HBcAg 83-144 aa were PCR-amplified, and then cloned into pcDNA3 plasmid. The chimeric gene was subcloned into the prokaryotic vector, pRSET-B.

Interaction of hepatitis C virus envelope glycoprotein E2 with the large extracellular loop of tupaia CD81.

Aim: To further analyze the interaction of tupaia CD81 with hepatitis C virus (HCV) envelope protein E2.

Methods: A tupaia CD81 large extracellular loop (CD81 LEL), which binds to HCV E2 protein, was cloned and expressed as a GST-fusion protein, and interaction of HCV E2 protein with a tupaia CD81 LEL was evaluated by enzyme-linked immunosorbent assay (EIA).

Results: Although tupaia and human CD81 LEL differed in 6 amino acid changes, tupaia CD81 LEL was strongly recognized by anti-CD81 antibodies against human CD81 LEL conformation-dependent epitopes.

[Prokaryotic expression and polyclonal antibody preparation of truncated duck hepatitis B virus core protein].

Aim: To construct a prokaryotic plasmid expressing truncated duck hepatitis B virus core protein (DHBc(1-214)), purify the recombinant protein, and to develop polyclonal antibodies against DHBc.

Methods: DHBc(1-214) was cloned into vector pRSET-B, then expressed in E.coli Rosetta(DE3) pLacI induced by IPTG.

[Genetic evaluation of Toxicodendron vernicifluum cultivars using amplified fragment length polymorphism markers].

Accurate identification of Toxicodendron vernicifluum varieties and understanding their genetic relationships are essential for the improved varieties selection. In this study, Amplified fragment length polymorphism (AFLP) markers was used to evaluate the genetic resources of nine Toxicodendron vernicifluum cultivars from Shaanxi. Eight pairs of AFLP primers with Mse I fluorescent labeled were screened from EcoR I /Mse I primer combinations.

Expression of PTEN, PPM1A and P-Smad2 in hepatocellular carcinomas and adjacent liver tissues.

Aim: To investigate the expressions of PTEN, PPM1A and P-Smad2 in hepatocellular carcinoma (HCC) and their significance.

Methods: The expressions of PTEN, PPM1A and P-Smad2 in 31 HCC tissues, 25 adjacent liver tissues and 13 non-tumor liver tissues were detected by using Envision immunohistochemical technique.

Results: The positive expression (64.

N-terminal and C-terminal cytosine deaminase domain of APOBEC3G inhibit hepatitis B virus replication.

Aim: To investigate the effect of human apolipoprotein B mRNA-editing enzyme catalytic-polypeptide 3G (APOBEC3G) and its N-terminal or C-terminal cytosine deaminase domain-mediated antiviral activity against hepatitis B virus (HBV) in vitro and in vivo.

Methods: The mammalian hepatoma cells HepG2 and HuH7 were cotransfected with APOBEC3G and its N-terminal or C-terminal cytosine deaminase domain expression vector and 1.3-fold-overlength HBV DNA as well as the linear monomeric HBV of genotype B and C.

Inhibition of hepatitis B virus replication by APOBEC3G in vitro and in vivo.

Aim: To investigate the effect of APOBEC3G mediated antiviral activity against hepatitis B virus (HBV) in cell cultures and replication competent HBV vector-based mouse model.

Methods: The mammalian hepatoma cells Huh7 and HepG2 were cotransfected with various amounts of CMV-driven expression vector encoding APOBEC3G and replication competent 1.3 fold over-length HBV.

[Cloning, prokaryotic expression and polyclonal antibody preparation of HCCR: a new biomarker for hepatocellular carcinoma].

Objectives: To construct a prokaryotic plasmid expressing truncated human cervical cancer oncogene (HCCR-1(167-360)), to express and purify the recombinant protein, and to develop the polyclonal antibody against HCCR.

Methods: HCCR-1(167-360) was amplified by RT-PCR from HepG2 cells and cloned into vector pRSET-B, then expressed in E.coli BL21(DE3) pLysS, which was induced by IPTG.

[Infection of tupaia hepatocytes with hepatitis C virus in vitro].

Objective: Tupaia belangeri (tree shrew) has a close phylogenetic relationship with primates and has been shown to be susceptible to a variety of human viruses. This study was conducted to investigate whether or not hepatitis C virus (HCV) could infect primary tupaia hepatocytes (PTHs) in vitro.

Methods: Serum-derived HCV was cultivated with PTHs, and then positive and negative strand HCV RNA in PTHs, as well as the encapsidated HCV RNA in the culture medium were detected to evaluate the infection.

[Distribution of hepatitis B virus genotypes in Hubei province and its clinical significance].

Objective: To investigate the distribution of hepatitis B virus genotype in Hubei province (China) and its clinical significance.

Methods: Serum samples from 190 HBV DNA positive patients with chronic HBV infection,including 52 asymptomatic HBV carriers (ASC), 56 chronic hepatitis (CH), 32 fulminant hepatic failure (FHF), 22 liver cirrhosis (LC), and 28 hepatocellular carcinoma (HCC) patients were collected and tested for HBV genotypes by type-specific primers.

Results: A simple and precise genotyping system based on PCR using type-specific primers was developed for the determination of genotypes of hepatitis B virus (HBV).

Alternation of AFP-mRNA level detected in blood circulation during liver resection for HCC and its significance.

Aim: To investigate whether the current manipulation of liver resection for hepatocellular carcinoma (HCC) causes dissemination of liver tumor cells into blood circulation.

Methods: Fifty patients with HCC, but without evidences of metastases, were enrolled for the study. Samples of peripheral blood before skin incision and after abdominal wall suture, and those of hepatic venous blood and portal venous blood after liver parenchyma dissection,were obtained.

Hepatitis C virus may infect extrahepatic tissues in patients with hepatitis C.

AIM:To explore the status of extrahepatic hepatitis C virus (HCV) infection and replication in hepatitis C patients,and its potential implication in HCV infection and pathogenicity.METHODS:By reverse-transcriptase poly-merase chain reaction (RT-PCR),in situ hybridization (ISH) and immunohis-tochemistry, HCV RNA, HCV replicative intermediate (minus-strand of HCV RNA), and HCV antigens were detected in 38 autopsy extrahepatic tissue specimens (including 9 kidneys, 9 hearts, 9 pancreas, 5 intestines, 2 adrenal glands, 2 spleens, 1 lymph node, and 1 gallbladder) from 9 hepatitis C patients, respectively; and the status of HCV replication in extrahepatic tissues was studied.RESULTS:By RT-PCR, all 9 patients were positive for HCV RNA in kidney, heart, pancreas, and intestine, but only 6(66.

Analysis of in vivo patterns of caspase 3 gene expression in primary hepatocellular carcinoma and its relationship to p21(WAF1) expression and hepatic apoptosis.

AIM:To detect the expression of caspase 3 gene in primary human hepatocellular carcinoma (HCC) and investigate its relationship top21(WAF1) gene expression and HCC apoptosis.METHODS:In situ hybridization was employed to determine caspase 3 and p21(WAF1) expression in HCC.In situ end-labeling was used to detect hepatocytic apoptosis in HCC.

FADD and TRADD expression and apoptosis in primary hepatocellular carcinoma.

AIM:To investigate the clinical features of FADD and TRADD expressions in primary hepatocellular carcinoma (HCC) and to determine their relationship with hepatic apoptosis.METHODS:FADD and TRADD expressions were detected by immunohis-tochemistry and hepatic apoptosis were determined by in situ end-labeling (ISEL).RESULTS:Ten (25.