1.Preliminary study on genotype of hepatitis B virus detected from Tibetans in China.
Yan-qing XU ; Yong-dong ZHOU ; Sheng-li BI
Chinese Journal of Experimental and Clinical Virology 2005;19(2):118-120
<b>OBJECTIVEb>To determine the main genotype of hepatitis B virus (HBV) detected from Tibetans in China and provide basic data for hepatitis control and prevention.
<b>METHODSb>The S gene and C gene were amplified by PCR from the sera of HBsAg positive Tibetans. After sequencing, the gene sequences were analyzed and the phylogenetic trees were drawn by the software MEGA3.
<b>RESULTSb>In trees based on S gene, the sequences of most samples clustered at genotype D, while in trees based on C gene, the sequences of all samples clustered at genotype C.
<b>CONCLUSIONb>The dominant genotype of HBV detected from Tibetans in China is a C/D hybrid.
Genotype ; Hepatitis B ; blood ; epidemiology ; virology ; Hepatitis B Core Antigens ; genetics ; Hepatitis B Surface Antigens ; blood ; genetics ; Hepatitis B virus ; classification ; genetics ; immunology ; Humans ; Phylogeny ; Tibet ; epidemiology
3.Replication and encapsidation of HBV mutants with the truncated C gene.
Ju-qiang HAN ; Da-rong HU ; Jin-hua XIONG ; Xue-ling HU ; Gong-ren FAN ; Juan LI ; Chao-ying LIU ; Yi-pin DI ; Yi-pin WU
Chinese Journal of Experimental and Clinical Virology 2004;18(1):39-42
<b>OBJECTIVEb>To evaluate the replication and encapsidation of HBV mutants with the truncated C gene.
<b>METHODSb>The HBV mutants with the truncated C gene were constructed by molecular cloning and PCR-based deletion in vitro. The replication and encapsidation of HBV mutants were investigated by Southern blotting, PCR and real-time fluorescence PCR respectively after transfecting the HBV mutants plasmid into HepG2 cells by using liposome.
<b>RESULTSb>The C-truncated HBV mutant vectors were constructed successfully and confirmed exactly by clone sequencing and enzymes digestion. The C-truncated HBV mutants were replication defective, however, all types of HBV DNA could be detected positive in the cytoplasm and supernatant after co-transfecting the C-truncated HBV mutants plasmid and the helper constructs into HepG2 cells. The C-truncated HBV mutants were proved to produce 3-40 folds more progeny DNA than that of the wild-type HBV by DNA quantitative assay.
<b>CONCLUSIONb>The C-truncated HBV mutants are replication-deficient and could not replicate and encapsulate in the hepatocytes when transfected solely, however, the progeny HBV-variant viruses are encapsidated more effectively to secrete into supernatant when co-transfected with the helper construct which lacks part of 5 prime-proximal HBV RNA packaging signal Epsilon.
Cell Line, Tumor ; Hepatitis B Core Antigens ; genetics ; Hepatitis B virus ; genetics ; physiology ; Humans ; Mutation ; Plasmids ; genetics ; Transfection ; Virus Replication
5.Construction and antigenic evaluation of a recombinant MVA virus-like particle expressing HBV C gene.
Xiang-ling LUAN ; Wei KONG ; Su-jun LIU ; Li LEI ; Yan HU ; Jun HOU ; Hong-hui SHEN ; Yi-chen WU ; Shao-li YOU ; Pan-yong MAO ; Shao-jie XIN
Journal of Southern Medical University 2008;28(2):252-254
<b>OBJECTIVEb>To construct the virus-like parcel expressing hepatitis B virus (HBV) C gene and identify its immunogenicity.
<b>METHODSb>HBV C gene was cloned into the shuttle vector pSC11, and the resulted plasmid pSC11-C was transfected into modified vaccinia virus Ankara (MVA).
<b>RESULTSb>pSC11-C was correctly constructed as verified by sequence analysis and PCR, and the recombinant virus-like parcel possessed good immunogenicity.
<b>CONCLUSIONb>The MVA-C expressing HBV C gene has been successfully constructed to provide important basis for gene therapy research of chronic HBV infection.
Genes, Viral ; Genetic Vectors ; Hepatitis B Core Antigens ; genetics ; Recombination, Genetic ; Vaccinia virus ; genetics
8.Relationship between basal core promoter combined point mutation of hepatitis B virus and TCM syndrome type.
Fei ZHOU ; Ling-tai WANG ; Jian-jie CHEN ; Gang ZHAO ; Bin ZHANG
Chinese Journal of Integrated Traditional and Western Medicine 2005;25(9):777-779
<b>OBJECTIVEb>To investigate the relationship between basal core promoter (BCP) combined point mutation of hepatitis B virus (HBV) and TCM syndrome type.
<b>METHODSb>One hundred and two patients with chronic hepatitis with positive HBV DNA and hadn't ever been treated by Lamivudine and interferon were differentiated according TCM syndrome differentiation into 5 types, two excess types (damp-heat blocking zhong-jiao type and blood stasis blocking collaterals type) and three deficiency types, gan-stagnation with pi-dificiency type, gan-shen yin-deficiency type and pi-shen yang-deficiency type. The serum HBV DNA, hepatic biochemical indexes, and the mutation of BCPnt 1762A-T and nt1764G-A combined point were determined, respectively.
<b>RESULTSb>The variant strain positive rate detected in the excess type was significantly higher than that in the deficiency type, the highest rate appeared in patients of damp-heat blocking zhong-jiao type.
<b>CONCLUSIONb>BCP combined point mutation may be liable to happen in patients of TCM excess type, especially in patients of damp-heat blocking zhong-jiao type.
Diagnosis, Differential ; Hepatitis B Core Antigens ; genetics ; Hepatitis B virus ; genetics ; Hepatitis B, Chronic ; diagnosis ; virology ; Humans ; Medicine, Chinese Traditional ; Point Mutation ; Promoter Regions, Genetic ; genetics
9.The influence of pre-core and BCP mutations on the severity of chronic hepatitis B.
Peng-Jian WENG ; Guo-Sheng GAO ; Shi-Xiong DING ; Xiao-Yue LIANG ; Xiang-Rong TANG
Chinese Journal of Hepatology 2006;14(10):769-771
Adolescent
;
Adult
;
Aged
;
DNA, Viral
;
Female
;
Hepatitis B Core Antigens
;
genetics
;
Hepatitis B virus
;
genetics
;
Hepatitis B, Chronic
;
genetics
;
Humans
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Male
;
Middle Aged
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Mutation
;
Young Adult
10.Study of the quasispecies dynamics of serum hepatitis B virus in a patient with acute exacerbations of chronic hepatitis B.
Lin LIU ; Yu-ming WANG ; Lin LAN ; Jun-gang LI ; Guo-hong DENG ; Jie XIA ; Xue-lan ZHAO ; Xu-qing ZHANG
Chinese Journal of Hepatology 2006;14(1):29-32
<b>OBJECTIVESb>To investigate the quasispecies dynamics of hepatitis B virus (HBV) during the course of exacerbation and resolution of chronic hepatitis B in a patients.
<b>METHODSb>Five serum samples were collected from a patient with two episodes of exacerbation and resolution of chronic hepatitis B. A method of PCR-TA cloning-conformation sensitive gel electrophoresis (CSGE)-sequencing was performed to study the dynamic changes of HBV quasispecies in basal core promoter (BCP), precore and core regions of HBV genome.
<b>RESULTSb>Quasispecies complexity was 10 and 12 at the points of exacerbations, and 14 and 17 at the points of resolutions (t = 3.133, P < 0.05). Ratio of dominant quasispecies in HBV population was high (42.4% and 51.5%) during exacerbations and low (30.3% and 21.2%) during resolutions (t = 3.295, P < 0.05). All dominant quasispecies, except the one during the second resolution, carried core P5T, L60V, S155T, and precore G1896A mutations.
<b>CONCLUSIONb>The composition of HBV quasispecies changes due to the change of host immune status, and immune pressure might lead to the selection of immune escape mutants.
Adolescent ; Hepatitis B Core Antigens ; genetics ; Hepatitis B virus ; classification ; genetics ; Hepatitis B, Chronic ; immunology ; virology ; Humans ; Male ; Mutation ; Promoter Regions, Genetic ; genetics