DNA, Circular ; isolation & purification ; DNA, Viral ; isolation & purification ; Hepatitis B ; diagnosis ; Hepatitis B virus ; genetics ; Humans

China ; epidemiology ; Hepatitis B ; classification ; epidemiology ; virology ; Hepatitis B Surface Antigens ; blood ; Hepatitis B virus ; genetics ; isolation & purification ; Humans ; Mutation

OBJECTIVESTo study the phenomena of hepatitis B virus (HBV) integration into the tissues of hilar cholangiocarcinoma (HCCA) and to identify the integration sites in the host genome.

METHODSTen fresh HCCA samples were collected from the tissues by surgical ablation, 1 normal hilar bile duct sample selected as control. Cellular DNA were extracted by Wizard SV Genomic DNA Purification System. PCR-derived assay (HBV-Alu-PCR) was employed to amplify the viral-host junctions which contain the HBV sequence and the adjacent cellular flanking sequences. The PCR products were purified and subjected to sequencing by ABI-3730XL Auto DNA Analyzer. The sequence analysis of viral-host junctions was performed by DNASIS MAX 3.0 bioinformatics software. The insertion sites between viral and cellular sequences were identified through homology comparison using NCBI BLAST and MapViewer search.

RESULTSIn 10 HCCA samples, 5 were demonstrated to have HBV integration fragments with total 6 inserted sites identified. Sequence analysis from viral-host junction showed that HBV X gene inserted into host genome at random distribution with truncated fragments. HBV integration recurrently targeted the unknown region in upstream of CXXC finger protein-1 (CpG-binding protein) gene (4 cases). p53 tumor suppressor gene was also found at the integration site.

CONCLUSIONSThere is high integration rate of HBV DNA into cellular genome of HCCA. HBV integration is found frequently into or close to cancer-related genes. The findings demonstrate that HBV infection might have association with the pathogenesis of HCCA.

Aged ; Base Sequence ; Bile Duct Neoplasms ; genetics ; virology ; Cholangiocarcinoma ; genetics ; virology ; DNA, Viral ; genetics ; Female ; Hepatitis B ; virology ; Hepatitis B virus ; genetics ; isolation & purification ; Humans ; Male ; Virus Integration

OBJECTIVETo investigate the presentation of a neutralization epitope-containing peptide antigen of hepatitis E virus (HEV) on chimeric virus-like particles (VLPs) of hepatitis B surface antigen (HBsAg).

METHODSThe gene fragment corresponding to amino acids (aa) 551-607 (HEnAg) of HEV capsid protein, which contains the only neutralization epitope identified to date, was fused via a synthetic glycine linker in frame with the gene of HBsAg. The resulted fusion gene was then integrated through transformation into the genome of Pichia pastoris under the control of a methanol-induced alcohol oxidase 1 (AOX1) promoter and expressed intracellularly. The expression products in the soluble cell extracts were characterized by Western blot, ELISA, CsCl density gradient analysis, and electron microscopic visualization.

RESULTSThe novel fusion protein incorporating HBsAg and the neutralization epitope-containing HEnAg was expressed successfully in Pichia pastoris with an expected molecular weight of approximately 32 kD. It was found to possess the ability to assemble into chimeric HBV/HEV VLPs with immunological physical and morphological characteristics akin to HBsAg particles. Not only did the chimeric VLPs show high activity levels in a HBsAg particle-specific ELISA but they were also strongly immunoreactive with hepatitis E (HE) positive human serum in a HEV specific ELISA, indicating that HEnAg peptide fragments were exposed on VLP surfaces and would be expected to be readily accessible by cells and molecules of the immune system. Similarity between chimeric VLPs to highly immunogenic HBsAg particles may confer good immunogenicity on surface-displayed HEnAg.

CONCLUSIONThe chimeric HBV/HEV VLPs produced in this study may have potential to be a recombinant HBV/HEV bivalent vaccine candidate.

Epitopes ; Genetic Engineering ; Hepatitis Antigens ; biosynthesis ; genetics ; immunology ; Hepatitis B Surface Antigens ; biosynthesis ; genetics ; immunology ; Hepatitis B virus ; genetics ; immunology ; Hepatitis E virus ; genetics ; immunology ; Pichia ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; isolation & purification ; Vaccines, Synthetic

OBJECTIVETo identify the intergenotype recombination of hepatitis B virus (HBV) in Chinese patients and explore new recombination types of HBV.

METHODSComplete genome sequences of HBV from Chinese patients were downloaded from GenBank and analyzed by Fragment-typing method to screen the potential recombinants, and the breakpoints of recombination were identified by using Simplot program.

RESULTSThirty-one recombination types including 755 recombinants were identified from 1642 complete HBV genome sequences, including 22 B/C recombination types with 676 recombinants, 5 C/D recombination types with 75 recombinants, 3 A/C recombination types with 3 recombinants, and 1 C/I recombination type with 1 recombinant.

CONCLUSIONOf the 31 recombination types comprising 755 recombinants, 4 are novel recombination types found for the first time. All these recombination types identified in this study involve genotype C.

Genome, Viral ; Genotype ; Hepatitis B virus ; genetics ; isolation & purification ; Humans ; Phylogeny ; Recombination, Genetic

OBJECTIVETo investigate whether the hepatitis B virus (HBV) has quasispecies character by studying nucleotide sequence polymorphism and mutation features of HBV PreC/C gene region, and preliminaryly explore the heterogeneity of HBV quasispecies.

METHODSThe serum sample was obtained from a patient with chronic hepatitis B, and the whole HBV PreC/C gene region was amplified by PCR and cloned. Thirty-four clones that contained HBV PreC/C gene fragments were sequenced.

RESULTSThere were 28 kinds of different nucleotide sequences in 34 clones, and the nucleotide sequences diversity ranged from 0.2% to 2.1%. The mutation points were almost distributed in the whole region, but there wasn't mutation at PreC region nt.1 896 point in all sequences.

CONCLUSIONHepatitis B virus has complex quasispecies character in the patients with chronic hepatitis B.

Adult ; Hepatitis B Core Antigens ; genetics ; immunology ; Hepatitis B virus ; classification ; genetics ; isolation & purification ; Hepatitis B, Chronic ; virology ; Humans ; Male ; Mutation ; Promoter Regions, Genetic ; genetics ; Sequence Analysis, DNA

This article describes the nomenclature, history and genetic evolution of duck hepatitis A virus, and updates the epidemiology, clinical symptom and surveillances of duck virus hepatitis A. It also summarizes the present status and progress of duck virus hepatitis A and illustrated the necessity and urgency of its research, which provides rationale for the control of duck hepatitis A virus disease in China.
Animals ; Ducks ; virology ; Hepatitis Virus, Duck ; classification ; genetics ; isolation & purification ; Hepatitis, Viral, Animal ; virology ; Picornaviridae Infections ; veterinary ; virology

Genes, Viral ; Genotype ; Hepatitis B virus ; genetics ; isolation & purification ; Hepatitis B, Chronic ; virology ; Humans ; Polymerase Chain Reaction ; methods

Adult ; China ; epidemiology ; Genes, Viral ; Genotype ; Hepatitis B ; epidemiology ; virology ; Hepatitis B virus ; classification ; genetics ; isolation & purification ; Humans ; Male

Animals ; China ; epidemiology ; Disease Reservoirs ; Hepatitis E ; epidemiology ; transmission ; Hepatitis E virus ; genetics ; isolation & purification ; Humans ; Rodentia ; virology ; Swine ; virology