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
2.Hepatitis B virus/human immunodeficiency virus coinfection: interaction among human immunodeficiency virus infection, chronic hepatitis B virus infection, and host immunity.
Yi-Jia LI ; Huan-Ling WANG ; Tai-Sheng LI
Chinese Medical Journal 2012;125(13):2371-2377
<b>OBJECTIVEb>This review discusses progress in the studies of hepatitis B virus (HBV)/human immunodeficiency virus (HIV) coinfection and focuses on the interaction among HIV infection, chronic HBV infection, and host immunity.
<b>DATA SOURCESb>Data and studies published mainly from 2008 to 2011 were selected using PubMed.
<b>STUDY SELECTIONb>Original articles and critical reviews concerning HBV/HIV coinfection and HBV and HIV pathogenesis were selected.
<b>RESULTSb>HIV may accelerate HBV progression by lowering CD4 count, weakening HBV-specific immunity, "enriching" HBV mutants, causing immune activation, etc. On the other hand, HBV may enhance HIV replication by activating HIV long terminal repeat (LTR) with X protein (HBX) and cause immune activation in synergy with HIV. Paradoxically, HBV may also inhibit HIV dissemination via dendritic cells.
<b>CONCLUSIONSb>The interaction among HIV, HBV, and host immunity remains poorly understood. Further research is warranted to elucidate the detailed molecular mechanisms and to translate these mechanisms into clinical practice.
HIV ; pathogenicity ; HIV Infections ; immunology ; Hepatitis B virus ; pathogenicity ; Hepatitis B, Chronic ; virology ; Humans ; Immunity, Innate ; immunology
4.Heat impact upon the infectivity of hepatitis B virus in serum.
Xiu-xia SONG ; Li-wen JU ; Guo-rong WEI ; Qing-wu JIANG
Chinese Journal of Preventive Medicine 2011;45(8):723-726
<b>OBJECTIVEb>This article was to explore the impact of temperature on hepatitis B virus infectivity.
<b>METHODSb>HBV positive serum with a HBV DNA titer of 1.33 × 10(8) copies/ml was aliquots into 23 Ep tubes with 1.5 ml, 100 µl in one tube.15 tubes were incubated at 37, 56 and 65°C for 0, 30, 60, 120 and 600 minutes, respectively. The other 8 tubes were incubated at 98°C for 0, 5, 10 and 30 minutes, respectively. Post-treated serum at all time points were selected to infect HepG-2 cell. When 18 hours after infection, these cells were extensively washed with phosphate buffered saline. Cells were harvested after the addition of fresh culture medium to culture cells for 48 hours. HBV DNA was detected by FQ-PCR.
<b>RESULTSb>HBV DNA was detected in cells that were infected by serum at 37°C and 56°C for 30, 60, 120 and 600 minutes, respectively. The titers for the cells incubated at 37°C were (4.85 ± 1.71) × 10(5), (3.85 ± 1.76) × 10(5), (1.67 ± 0.67) × 10(5), (7.86 ± 1.03) × 10(4) copies/ml, and those for the cells incubated at 56°C were (4.01 ± 0.16) × 10(5), (9.77 ± 0.97) × 10(4), (6.36 ± 0.65) × 10(4), (5.05 ± 0.24) × 10(3) copies/ml at different incubation time points. For the cells incubated at 65°C for 60 and 120 minutes, HBV DNAs were (5.15 ± 7.28) × 10(3) and (7.56 ± 10.60) × 10(2) copies/ml, respectively, which were much lower than those in the controls cells ((6.79 ± 1.48) × 10(5) copies/ml). The results of HBV DNA were different (F = 104.4, P < 0.001) in groups treated with different temperature, and results of HBV DNA were also different (F = 144.0, P < 0.001) in groups processed for different period of time. Temperature and processing time had interaction (F = 23.6, P < 0.001). After heating at 98°C for 10 minutes and boiling for 5 minutes, the HBV DNA copy number ((3.02 ± 4.26) × 10(2), (4.31 ± 6.09) × 10(2) copies/ml) in infected cells decreased by about 10 folds than that in the control group ((6.79 ± 1.48) × 10(5) copies/ml). HBV DNAs were not detected in cells that were infected by serum which was heated at 98°C for 30 minutes and boiled for 10 minutes.
<b>CONCLUSIONb>The infectivity of HBV serum in vitro was relatively stable at low temperature, and it would lose its infectivity in short period of time at high temperature.
Hep G2 Cells ; Hepatitis B virus ; pathogenicity ; physiology ; Hot Temperature ; Humans ; Serum ; virology
5.Antiviral therapy of decompensated hepatitis B virus-related cirrhosis.
Guang-Cheng CHEN ; Tao YU ; Kai-Hong HUANG ; Qi-Kui CHEN
Chinese Medical Journal 2012;125(2):373-377
<b>OBJECTIVEb>To review the development, mechanism, necessity and limitation of antiviral therapy in decompensated hepatitis B virus-related cirrhosis.
<b>DATA SOURCESb>Most information was pulled from a literature search (Pubmed 2000 to 2011) using the keywords of antiviral and decompensated hepatitis B virus-related cirrhosis. Relevant book chapters were also reviewed.
<b>STUDY SELECTIONb>Well-controlled, prospective landmark studies and review articles on antiviral therapy in decompesated hepatitis B virus-related cirrhosis were selected.
<b>RESULTSb>Specific antiviral agents not only control viral replication, which permits liver transplantation, but also improve liver function so significantly that patients could be removed from the transplant waiting list. However, the emergence of drug-resistant mutants can result in treatment failure. Combination therapy is a save-strategy in drug-resistant.
<b>CONCLUSIONSb>Although the treatment of end-stage liver disease is still a challenge worldwide, antiviral therapy has altered the natural history of hepatitis B patients with decompensated cirrhosis. The approval of the new generation of antivirals is opening new perspectives for finding the optimal antiviral treatment for patients with decompensated cirrhosis and preventing antiviral resistance. A combination of antivirals may be one of the future strategies for fulfilling these goals.
Antiviral Agents ; therapeutic use ; Hepatitis B virus ; drug effects ; pathogenicity ; Humans ; Liver Cirrhosis ; drug therapy ; virology
6.Studies on the association between beta 2-glycoprotein I and hepatotropism of hepatitis B virus.
Yan-hang GAO ; Pu-jun GAO ; Dan WANG ; Yang SHI ; Tong-yu TANG ; Yun-feng PIAO ; Han-yi YANG
Chinese Journal of Hepatology 2006;14(8):569-571
<b>OBJECTIVEb>To further study the binding character of hepatitis B surface antigen (HBsAg) and beta 2-glycoprotein I (beta2GP I) and to explore whether beta2GP I plays an important role in the hepatotropism of hepatitis B virus.
<b>METHODSb>Using Western blot technique, we observed the binding character of the HBsAg with reduced and non-reduced beta2GP I.
<b>RESULTSb>rHBsAgs with reduced and non-reduced beta2GP I showed identical binding activity.
<b>CONCLUSIONSb>The binding activity of HBsAg is dependent on tandem residues, but not on conformational structures of beta2GP I. There is a specific binding between HBV and beta2GP I, which may play an important role in HBV infection and is one of the reasons of hepatotropism of HBV.
Hepatitis B ; virology ; Hepatitis B Surface Antigens ; metabolism ; Hepatitis B virus ; pathogenicity ; Humans ; Viral Envelope Proteins ; blood ; beta 2-Glycoprotein I ; blood
7.The analysis to the serological epidemiology of hepatitis virus of Fangshan district in Beijing.
Hai-Zhu YU ; Na XIANG ; Lan-Mei CUI
Chinese Journal of Epidemiology 2008;29(2):205-205
Adolescent
;
Adult
;
Age Distribution
;
Child
;
Child, Preschool
;
Female
;
Hepatitis A
;
epidemiology
;
virology
;
Hepatitis A Virus, Human
;
pathogenicity
;
Hepatitis B
;
epidemiology
;
virology
;
Hepatitis B virus
;
pathogenicity
;
Humans
;
Infant
;
Infant, Newborn
;
Male
;
Middle Aged
;
Socioeconomic Factors
;
Young Adult
9.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
10.Detection of carboxyhemoglobin in patients with hepatic encephalopathy due to hepatitis B virus-related cirrhosis.
Xiao-yu SUN ; Zhi-jun DUAN ; Yan-lian LI ; Qing-shan CHANG
Chinese Medical Journal 2012;125(22):3991-3996
<b>BACKGROUNDb>The heme oxygenase/carbon monoxide (HO/CO) system plays an important role in the development of hepatic fibrosis. The level of the HO/CO can be directly obtained by determining the carboxyhemoglobin (COHb) level. The aims of this study were to reveal the significance of COHb in patients with hepatitis B virus-related cirrhosis (HBC) complicated by hepatic encephalopathy (HE), and to further investigate the influence of the HO/CO pathway on the end-stage cirrhosis, hoping to find a reliable indicator to evaluate the course of HBC.
<b>METHODSb>According to the diagnostic criteria, 63 HBC inpatients with HE were enrolled in group H. Patients regaining awareness with current therapies were categorized into group P-H. Comparisons were made with a control group (group N) consisting of 20 health volunteers. The levels of COHb, partial pressure of oxygen (PaO2) and oxygen saturation (SaO2) were determined by arterial blood gas analysis method. The incidences of hepatorenal syndrome (HRS), upper gastrointestinal bleeding, esophagogastric varices and spontaneous bacterial peritonitis (SBP) in group H were recorded. COHb levels in different groups were compared, and the correlations of COHb levels with HE grades (I, II, III, and IV), PaO2, SaO2 and hypoxemia were analyzed.
<b>RESULTSb>The COHb level in group P-H ((1.672 ± 0.761)%) was significantly higher than that in group N ((0.983 ± 0.231)%) (P < 0.01), and the level in group H ((2.102 ± 1.021)%) was significantly higher than groups P-H and N (P < 0.01). A positive correlation was observed between the COHb concentration and the grade of HE (r(s) = 0.357, P = 0.004). There were no significant differences of COHb levels between HE patients with and without complications such as esophagogastric varices ((2.302 ± 1.072)% vs. (1.802 ± 1.041)%, P > 0.05) or the occurrence of SBP ((2.960 ± 0.561)% vs. (2.030 ± 1.021)%, P > 0.05). Compared with HE patients with HRS, the level of COHb was significantly higher in HE patients without HRS ((2.502 ± 1.073)% vs. (1.981 ± 1.020)%, P = 0.029). The COHb level had a negative correlation with PaO2 (r = -0.335, P = 0.007) while no statistically significant relationship was found with SaO2 (r = -0.071, P > 0.05). However, when the above two parameters met the diagnostic criteria of hypoxemia, the COHb concentration increased ((2.621 ± 0.880)% vs. (1.910 ± 0.931)%, P = 0.011).
<b>CONCLUSIONSb>COHb is a potential candidate to estimate the severity and therapeutic effect of HE. The levels of COHb may be tissue-specific in cirrhotic patients with different complications.
Adult ; Aged ; Carboxyhemoglobin ; metabolism ; Female ; Fibrosis ; complications ; virology ; Hepatic Encephalopathy ; metabolism ; Hepatitis B virus ; pathogenicity ; Humans ; Male ; Middle Aged