1.Establishment of a semi-nested PCR for identifying the sub-genotypes (Ba and Bj) of hepatitis B virus of genotype B.
Jie WANG ; Jun-Wei GAO ; Jie LI ; Hui ZHUANG ; Jia WANG ; Ya-Juan LI ; Hui JIN
Chinese Journal of Epidemiology 2008;29(2):177-180
<b>OBJECTIVEb>To establish a sensitive, specific, simple and practicable method for identifying the two sub-genotypes (Ba and Bj) of genotype B isolates of hepatitis B virus (HBV) (HBV/ B).
<b>METHODSb>The entire nucleotide sequences of HBV/B and HBV/C were obtained from GenBank, compared and analyzed with DNAStar software. The specific primers for HBV/B (HB) and Ba (BA), Bj (BJ) were designed respectively. HB as HBV/B specific primer (sense) and HBAS-4V (designed by Japanese scientists Sugauchi et al) as a universal outer primer (antisense) were used in the first-round PCR. In the second-round PCR, HB was also used as sense primer while BA and BJ as inner primers (antisense) and they were added into a single tube for PCR reaction. The two sub-genotypes of HBV/B were identified according to the length of the PCR products. A total of 71 HBV DNA-positive serum samples were selected randomly from our laboratory, including 56 HBV/B samples identified by type-specified PCR method and 15 HBV/C samples identified by direct sequencing in preS and S Region (preS/S). All the 71 samples were detected with this semi-nested PCR method. A plasmid containing full length genomic DNA of HBV/Bj, was presented by Professor Kenji Abe as positive control of Bj. Then, the first-round PCR products of 15 HBV/B were randomly selected and sequenced directly, and their sequences were compared phylogenetically with the above known Ba and Bj sequences using Blast and DNAStar softwares to confirm the results of semi-nested PCR.
<b>RESULTSb>56 HBV/B samples were all identified as HBV/Ba by our semi-nested PCR method. 15 randomly selected PCR products were all sequenced as HBV/Ba. All of the 15 HBV/C samples were negative.
<b>CONCLUSIONb>A simple, rapid, sensitive and specific method for identifying sub-genotypes Ba and Bj was established with might be used for large-scale clinical and epidemiological studies.
Genotype ; Hepatitis B ; virology ; Hepatitis B virus ; classification ; genetics ; pathogenicity ; Humans ; Phylogeny ; Polymerase Chain Reaction ; methods
3.Advancement of researches on relationship between hepatitis B virus and severe hepatitis B.
Journal of Biomedical Engineering 2009;26(4):904-907
Approximately 1% of patients infected with hepatitis B virus (HBV) develops severe hepatitis B, which is characterized by rapid destruction of liver parenchyma. The case fatality rates of HBV-induced severe hepatitis B in China range between 60% and 80%. In other words, more than half of the patients with severe hepatitis B ultimately require liver transplantation or die of liver failure. The pathogenesis of severe hepatitis B is not well understood. Previous studies have indicated that complicated interaction between viral factors and host immune response may be involved. This article tries to review the advancement of researches on the relationship between HBV and severe hepatitis B.
Hepatitis Antibodies
;
blood
;
Hepatitis B
;
immunology
;
virology
;
Hepatitis B virus
;
genetics
;
pathogenicity
;
Host-Pathogen Interactions
;
genetics
;
Humans
;
Mutation
;
Severity of Illness Index
5.Molecular Epidemiology of Hepatitis B Virus.
The Korean Journal of Internal Medicine 2011;26(3):255-261
Although safe and effective vaccines for hepatitis B virus (HBV) have been available for nearly three decades, this virus kills at least 600,000 people annually worldwide and remains the leading global cause of chronic hepatitis, cirrhosis, and hepatocellular carcinoma. Because the HBV reverse transcriptase lacks a proofreading function, many HBV genotypes, subgenotypes, mutants, and recombinants exist. At least 10 HBV genotypes (HBV-A through J) with distinct geographic distributions have been identified; by definition, their complete genomic sequences diverge by more than 8%. HBV genotype is increasingly becoming recognized as an important factor in the progression and clinical outcome of HBV-induced disease. Infections by HBV-C or -D are significantly more likely to lead to cirrhosis and hepatocellular carcinoma than are infections by HBV-A or -B. Additionally, the hepatitis B e antigen seroconversion response to standard or pegylated interferon is more favorable in patients with HBV-A or -B than in those with HBV-C or -D. However, therapeutic responses to nucleos(t)ide analogues are generally comparable among HBV genotypes. In conclusion, genotyping of HBV is useful in identifying chronic hepatitis B patients who are at increased risk of disease progression, thereby enabling physicians to optimize antiviral therapy for these patients.
Antiviral Agents/therapeutic use
;
Drug Resistance, Viral/genetics
;
Genotype
;
Hepatitis B/diagnosis/drug therapy/*epidemiology
;
Hepatitis B virus/drug effects/*genetics/pathogenicity
;
Humans
;
Molecular Epidemiology
;
Phenotype
;
Prognosis
6.The association of HLA-DRB1 allele polymorphism with the genetic susceptibility to liver cirrhosis due to hepatitis B virus.
Yuan-qiao CHENG ; Ju-sheng LIN ; Li-hong HUANG ; De-ying TIAN ; Ping XIONG
Chinese Journal of Medical Genetics 2003;20(3):247-249
<b>OBJECTIVEb>To assess the association of human leucocyte antigen (HLA)-DRB1 allele with the genetic susceptibility to cirrhosis due to hepatitis B virus(HBV).
<b>METHODSb>One hundred and six patients with cirrhosis due to HBV in Hubei area were investigated for HLA-DRB1 gene by polymerase chain reaction-sequence specific primers technique. The results were compared with those from 108 normal healthy people.
<b>RESULTSb>The frequency of HLA-DRB1*1201/1202 allele was 20.28% in patient group, which was significantly higher than the frequency (6.01%) in control group, the relative risk (RR) being 4.9878 (P<0.01). The frequency of HLA-DRB1*1501/1502 allele was decreased in patient group (patient 6.6%, control 16.67%, RR=0.3043, P<0.05), while the frequencies of other HLA-DRB1 alleles were not significantly different(P>0.05).
<b>CONCLUSIONb>HLA-DRB1*1201/1202 allele may be the susceptibility gene in patients with cirrhosis due to HBV in Hubei Han nationality; HLA-DRB1*1501/1502 allele is a resistant gene in patients with cirrhosis.
Adult ; Female ; Genetic Predisposition to Disease ; HLA-DR Antigens ; genetics ; HLA-DRB1 Chains ; Hepatitis B ; complications ; genetics ; Hepatitis B virus ; genetics ; pathogenicity ; Hepatitis B, Chronic ; complications ; genetics ; Humans ; Liver Cirrhosis ; etiology ; genetics ; Male ; Middle Aged ; Polymorphism, Genetic
7.Association of-238G/A and -857C/T polymorphisms of tumor necrosis factor-alpha gene promoter region with outcomes of hepatitis B virus infection.
Hong-Quan LI ; Zhuo LI ; Ying LIU ; Jun-Hong LI ; Jian-Qun DONG ; Ji-Rong GAO ; Chun-Yan GOU ; Hui LI
Biomedical and Environmental Sciences 2006;19(2):133-136
<b>OBJECTIVESb>To determine whether -238G/A and -857C/T polymorphisms of tumor necrosis factor-alpha (TNF-alpha) gene promoter were associated with outcomes of hepatitis B virus infection.
<b>METHODSb>A total of 246 HBV self-limited infected subjects and 443 chronic hepatitis B (HB) patients were recruited in this case-control study. TNF-alpha-238G/A and -857C/T gene promoter polymorphisms were examined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP).
<b>RESULTSb>The frequency of TNF-alpha-238 GG (90.7%) in chronic HB group was significantly lower than that (95.1%) in self-limited group (P = 0.041). The frequency of TNF-alpha-857 CC (79.7%) in chronic HB patients was significantly higher than that (70.9%) in self-limited infected subjects (P = 0.021). Multiple logistic regression analysis revealed that both TNF-alpha-238GA and -857CC were independently associated with chronic HB.
<b>CONCLUSIONSb>TNF-alpha promoter variants are likely to play a substantial role in influencing the outcomes of HBV infection.
Adult ; Case-Control Studies ; Female ; Haplotypes ; Hepatitis B ; genetics ; physiopathology ; Hepatitis B virus ; pathogenicity ; Hepatitis B, Chronic ; genetics ; physiopathology ; Humans ; Male ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Polymorphism, Restriction Fragment Length ; Promoter Regions, Genetic ; genetics ; Tumor Necrosis Factor-alpha ; genetics
8.Molecular Mechanisms of Hepatitis B Virus-associated Hepatocellular Carcinoma.
Neung Hwa PARK ; Young Hwa CHUNG
The Korean Journal of Hepatology 2007;13(3):320-340
Hepatocellular carcinoma (HCC) is one of the most common malignant diseases in the world. The hepatitis B virus (HBV) replicates non-cytopathically in hepatocytes, and most of the liver injury associated with this infection reflects the immune response. Epidemiological studies have clearly demonstrated that a chronic HBV infection is a major etiological factor in the development of HCC. The pathogenesis of HBV-associated HCC has been studied extensively, and the molecular changes during the malignant transformation have been identified. The main carcinogenic mechanism of HBV-associated HCC is related to the long term-inflammatory changes caused by a chronic hepatitis B infection, which might involve the integration of the HBV. Integration of the HBV DNA into the host genome occurs at the early steps of clonal tumorous expansion. The hepatitis B x protein (HBx) is a multifunctional regulatory protein that communicates directly or indirectly with a variety of host targets, and mediates many opposing cellular functions, including its function in cell cycle regulation, transcriptional regulation, signaling, encoding of the cytoskeleton and cell adhesion molecules, as well as oncogenes and tumor suppressor genes. Continued study of the mechanisms of hepatocarcinogenesis will refine our current understanding of the molecular and cellular basis for neoplastic transformations in the liver. This review summarizes the current knowledge of the mechanisms involved in HBV-associated hepatocarcinogenesis.
Apoptosis/genetics
;
Carcinoma, Hepatocellular/pathology/*virology
;
Cell Cycle
;
DNA Mismatch Repair
;
Hepatitis B virus/genetics/*pathogenicity
;
Hepatitis B, Chronic/*complications
;
Humans
;
Liver Neoplasms/pathology/*virology
;
Neovascularization, Pathologic/genetics/virology
;
Telomere/genetics
;
Trans-Activators/*metabolism
;
Transcription, Genetic
9.Relationship between amount of HBV DNA in serum/liver tissue and hepatitis G virus (HGV) infection in patients with chronic hepatitis B.
Qinghua SHANG ; Jianguo YU ; Hailong ZHUO ; Chuanzhen XU ; Ning WANG ; Guangshu ZHANG
Chinese Journal of Experimental and Clinical Virology 2002;16(4):326-328
<b>OBJECTIVEb>To observe the relationship between the amount of HBV DNA in serum/liver tissue and HGV infection in patients with chronic hepatitis B (CH-B) for exploring the effect of HGV infection on hepatitis B virus (HBV) replication of CH-B.
<b>METHODSb>HGV RNA in serum, HGV nonstructural region 5 (NS5) antigen (HGV Ag) in liver tissue and the amount of HBV DNA in serum, liver tissue were detected for 56 patients with CH-B by reverse transcription-polymerase chain reaction (RT-PCR) assay, peroxidase antiperoxidase (PAP) immunohistochemical method and fluorescence quantitative PCR assay, respectively. Then the relationship between HGV Ag expression in liver tissue and HGV RNA expression in serum was analysed and the amount of HBV DNA in serum and liver tissues from the serum HGV RNA or liver tissue HGV Ag positive patients were compared with those of the serum HGV-RNA or liver tissue HGV Ag negative patients, respectively.
<b>RESULTSb>Ten (17.9%) and eight (14.3%) patients were positive for serum and liver tissues,respectively.HGV RNA expression in serum was closely related to HGV Ag expression in liver tissues, but there was HGV RNA in serum from some of the liver tissues HGV Ag negative patients ?cases of HGV RNA and HGV Ag positive or negative,HGV RNA positive but HGV Ag negative, HGV RNA negative but HGV Ag positive, respectively: 5,43,5,3,(P<0.01). There was no significant difference in the amount of HBV DNA in serum and liver tissues between HGV RNA or HGV Ag positive and negative patients (P>0.05).
<b>CONCLUSIONSb>HGV infection may not affect HBV replication. Liver is the site of HGV replication, but HGV probably also replicates in extrahepatic tissues. HGV hepatic pathogenicity is probably mild and further studies are still needed.
Adult ; DNA, Viral ; analysis ; blood ; Female ; Flaviviridae Infections ; complications ; virology ; GB virus C ; genetics ; immunology ; pathogenicity ; Hepatitis Antigens ; analysis ; Hepatitis B virus ; genetics ; physiology ; Hepatitis B, Chronic ; complications ; virology ; Hepatitis, Viral, Human ; virology ; Humans ; Liver ; virology ; Male ; RNA, Viral ; blood ; Virus Replication
10.Efficacy and Safety of Tenofovir Disoproxil Treatment for Chronic Hepatitis B Patients with Genotypic Resistance to Other Nucleoside Analogues: A Prospective Study.
Jing ZHOU ; Yue-Ying LIU ; Jiang-Shan LIAN ; Li-Fang PAN ; Jian-Le YANG ; Jian-Rong HUANG
Chinese Medical Journal 2017;130(8):914-919
<b>BACKGROUNDb>Tenofovir disoproxil (TDF) is a promising salvage therapy for patients with chronic hepatitis B (CHB) who failed regimens of other nucleoside analogues (NAs). In this study, we aimed to investigate the clinical efficacy and safety of TDF monotherapy in Chinese CHB patients with genotypic resistance.
<b>METHODSb>A total of 33 CHB patients who had failed treatment with other NAs and had genotypic resistance were switched to TDF monotherapy for 48 weeks. Patients' demographic data (age, sex, history of hepatitis B virus [HBV] therapy), laboratory testing results (hepatitis B e antigen [HBeAg] status, HBV DNA levels, alanine aminotransferase [ALT] levels, serum creatinine, urinary protein, genotypic assay), clinical symptoms, and liver color ultrasound examinations were collected for evaluation at day 0 (baseline) and the 12th, 24th, 36th, and 48th weeks after initiating treatment. Statistical analyses were carried out using rank sum test or rank correlation.
<b>RESULTSb>With regard to efficacy, the study found that all patients who switched to TDF monotherapy had undetectable HBV DNA levels after 48 weeks. In addition, patients with lower baseline HBV DNA levels realized earlier virological undetectability (rs = 0.39, P = 0.030). ALT levels were normal in 30 of 33 patients (91%). HBeAg negative conversion occurred in 7 of 25 patients (28%), among whom HBeAg seroconversion (12%) and HBeAg seroclearance (16%) occurred. The time of complete virological response was significantly affected by the number of resistance loci (rs = 0.36, P = 0.040). Concerning safety, the study found that no adverse events were observed during the 48 weeks.
<b>CONCLUSIONb>TDF monotherapy is an effective and safe salvage treatment for CHB patients who are resistant to other NAs.
Adult ; Anti-HIV Agents ; adverse effects ; therapeutic use ; DNA, Viral ; genetics ; Drug Resistance, Viral ; Female ; Genotype ; Hepatitis B virus ; drug effects ; pathogenicity ; Hepatitis B, Chronic ; drug therapy ; Humans ; Male ; Middle Aged ; Prospective Studies ; Tenofovir ; adverse effects ; therapeutic use