Objective To investigate hepatic histological features and its influencing factors of HBeAg-negative chronic hepatitis B patients. Methods 134 HBeAg-negative chronic hepatitis B (CHB) patients who underwent percutaneous liver biopsy were recruited in this study. The liver biopsy sections were examined after routine haematoxylin-eosin (HE) staining, and silver staining for assessment of fibrosis. The activity of liver disease was assessed by using a modified Knodell numeric histology activity index (HAI). ALT level, HBV DNA load, HBV serological markers, HBV genotype were assessed with appropriate methods. t test or analysis of variance was used to compare means. Non-parametric was done by Kruskal-Wallis test. The correlation between liver pathological change and clinical factors was analyzed by multivariate linear regression. Results Of 134 HBeAg negative CHB patients, percentages of mild (HAI 4 ~ 8), moderate (HAI 9 ~ 12), and severe hepatitis (HAI 13 ~ 18) were 26.9%, 26.1%, and 47.0%, respectively. As for hepatic fibrosis, 18.7% and 81.3% of the patients had fibrosis score < 3 and ≥3, respectively. Multivariate regression analysis showed that ALT level and hepatic fibrosis were correlated to hepatic inflammation (t = 6.687,P < 0.01; t = 3.478, P < 0.01) while age and hepatic inflammation activity were influencing factors of hepatic fibrosis (t = 3.587, P < 0.01; t =7.136, P < 0.01). However, correlation is not significant between hepatic histological change and other factors, including gender, HBVDNA, HBV genotype and HBeAb status. Conclusions In this study, hepatic histological change tend to become worse in majority of HBeAg-negative chronic hepatitis B , especially in older patients and those with high ALT level.

Objective To investigate the efficacy of individualized interferon (IFN)-alpha therapy in HBeAg-negative chronic hepatitis B patients. Methods Seventy- six Chinese HBeAg-negative chronic hepatitis B patients proven by liver biopsy were treated with 5 MU recombinant IFN-alpha 1b subcutaneously thrice every week. All the patients were followed up for at least 24 months the combined responses were defined as normalization of serum alanine transaminase (ALT) and HBV DNA＜3 log10 copy/mL. An intention-to-treat (ITT) analysis was used in this paper in which all 76 patients were included. Results Six patients were lost. Treatment duration was in the range 2-24 months with a median of 8.5 months, and combined responses were achieved at a median of 6.0 months (range 2-19 months) of treatment duration.Seventy-five-percentile of treatment duration to endpoints was 10.0 months. The combined response rate was 46.1% (35/76) at the end of treatment, 43.3% (33/76) at 12-month follow-up and 40.8% (31/76) at 24-month follow-up. The relapse rate was 20. 0% (7/35) and 25. 7% (9/35) at 12-month and 24-month follow-up, respectively. Higher necroinflammatory activity in liver biopsy predicted a good response, while gender, age, liver fibrosis, baseline ALT, aspartate aminotransferase levels and baseline HBV DNA levels were not impact factors of therapeutic effects by binary Logistic regression analysis.Conclusion Individualized prolonged IFN-alpha regimen lead to considerable sustained disease suppression in patients with HBeAg-negative chronic hepatitis B.

OBJECTIVETo study the clinical and histological features of chronic hepatitis B (CHB) with negative hepatitis B e-antigen (HBeAg).

METHODSA total of 743 in-patients with chronic hepatitis B were recruited into the study and divided into two groups according to the HBeAg status. The correlation among alanine transaminase (ALT) levels, hepatitis B virus (HBV) DNA semiquantification, and the liver histopathological data were detected.

RESULTSOf the 743 successive in-patients, 267 (35.9%) were HBeAg-negative. The HBDAG-negative group had significantly lower serologic HBV DNA levels (63.0% of < 100 pg/ml) vs HBeAg-positive (42.6%, P < 0.001), while more sever inflammation (58.1% of inflammatory scores of histological activity index (HAIinf > or = 9) vs HBeAg-positive group (46.0%, P < 0.001) and severe fibrosis (45.3% of fibrosis scores of histological activity index (HAIfib > or = 3) vs HBeAg-positive group (27.9%, P < 0.001) of liver histology. In HBeAg-positive patients, increasing ALI levels were significantly associated with high inflammation and fibrosis scores and low HBV DNA levels. However, it was not the case in the HBeAg-negative cases. In HBeAg-positive patients, 91.3% of them had HAIinf > or = 9 and 65.7% had HAIfib > or = 3 with HBV DNA > 100 pg/ml, while 8.2% of them had HAIinf > or = 9 and 12.3% had HAIfib > or = 3 with HBV DNA < 20 pg/ml, indicating an obverse correlation between HBV DNA levels and histology scores.

CONCLUSIONSAs regards clinical and histological background, the chronic HBeAg-negative hepatitis B is a different subpopulation from the HBeAg-positive counterpart.

DNA, Viral ; Fibrosis ; pathology ; Hepatitis B ; immunology ; pathology ; Hepatitis B e Antigens ; analysis ; Hepatitis B virus ; genetics ; Hepatitis, Chronic ; Humans ; Liver ; pathology

OBJECTIVETo investigate the role of the 25 kD hepatitis B e antigen (HBeAg) precursor that only exist inside hepatocytes and study its effect on the pathopoiesis of hepatitis B and QIAGEN expression and purification system.

METHODSHepatitis B virus (HBV) preC/C gene for the 25 kD HBeAg precursor was cloned into the expression vector pQE30 and the 25 kD HBeAg precursor was expressed in Escherichia coli (E. coli) and purified. Its antigenicity for 21 kD mature HBeAg was tested by western blot analysis.

RESULTSCloned fragments in the expression vector were sequenced and verified to be homogeneous with that of HBV (ayw subtype). Expression of the HBeAg precursor in E. coli under the transcriptional regulation of T5 promoter yielded a soluble cytosolic protein with an apparent molecular mass of 25 kD. Recombinant HBeAg precursor exhibited identical potencies with 21 kD mature HBeAg that reacted with anti-HBeAg antibodies. The purification rate of the expressed HBeAg precursor was up to 89.6% and the yield of purified HBeAg precursor from this procedure was 2.4 mg/L.

CONCLUSION25 kD HBeAg precursor exhibited biological activity and might play an important role in pathopoiesis of hepatitis B.

Cloning, Molecular ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; genetics ; Gene Expression ; Hepatitis B e Antigens ; genetics ; isolation & purification ; metabolism ; Plasmids ; genetics ; Protein Precursors ; genetics ; isolation & purification ; metabolism ; Recombinant Proteins ; isolation & purification ; metabolism

OBJECTIVETo investigate the compositions of Th1/Th2/Th3 cells in chronic hepatitis B virus (HBV)-infected individuals by determining the expression of interleukin-4 (IL-4), inetrferon-gamma (IFN-gamma), and transform growth factor-beta (TGF-beta) in single CD4(+) T cells isolated from peripheral blood mononuclear cells (PBMCs) and the role of polarized Th cell populations in chronic HBV-infection was discussed.

METHODSPBMCs from chronically infected HBV individuals were isolated, stimulated by PMA/Ionomycin/Monensin, and IL-4, IFN-gamma and TGF-beta production by CD4(+) T cells was determined by using fluorescence activated cell sorter (FACS) analysis.

RESULTSThe percentage of IFN-gamma-producing T cells, IL-4-producing T cells and TGF-beta-producing T cells ranged from 2.3% - 18.6%, 1.1% - 8.7% and 0.7% - 7.1% respectively in CD4(+) T cells from non-infected individuals. Most of CD4(+) T cells from PBMCs in chronically infected HBV individuals were Th0 cells. The proportion of Th1 cells increased significantly with hepatic inflammatory activity, and in the active period of chronic hepatitis B infection were higher than those in the non-active period (P < 0.05). Th2 cell percentage in CD4(+) T cells from HBV-infected individuals did not differ significantly (P > 0.05), but were higher than that from controls (P < 0.05). Th3 cell percentage in CD4(+) T cells from asymptomatic carrier (AsC) group was higher than that in the chronic hepatitis B (CHB) and control groups (P < 0.05).

CONCLUSIONSTh1 phenotype cytokines were positively correlated with hepatic inflammatory activity in chronic hepatitis B and Th2 cells may be associated with the persistence of HBV infection. Th3 cells cooperating with Th2 cells can negatively regulate immune responses and may be associated with the immune tolerant state of chronic HBV infection.

CD4-Positive T-Lymphocytes ; immunology ; Hepatitis B, Chronic ; immunology ; metabolism ; pathology ; Humans ; Interferon-gamma ; biosynthesis ; Interleukin-4 ; biosynthesis ; T-Lymphocytes, Helper-Inducer ; immunology ; Th1 Cells ; immunology ; Th2 Cells ; immunology ; Transforming Growth Factor beta ; biosynthesis

OBJECTIVETo determine whether the transfusion-transmitted virus (TTV) is hepatotropic.

METHODSTotal DNA was extracted from various tissues of 5 experimentally infected Rhesus monkeys during the viremic period. A dot hybridization was done with viral double stranded DNA probes or single antisense probes.

RESULTSThe double-stranded probe was hybridized with DNA from the liver, bone marrow, spleen,stomach, small intestine and colon. The single-stranded antisense probe was hybridized with DNA from the liver, small intestine and bone marrow of all 5 monkeys, but not with that from other tissues.

CONCLUSIONSAs the viral genome was of negative polarity, the plus-stranded fragment identified in our study might be a replicative intermediate, and was only demonstrated in the liver, small intestine, and bone marrow by dot blot hybridization with single-stranded antisense probes. It is suggested that the TTV replicates in the liver, bone marrow and small intestine, and TTV might be hepatotropic.

Animals ; Bone Marrow ; virology ; DNA Virus Infections ; virology ; DNA, Viral ; analysis ; Intestine, Small ; virology ; Liver ; virology ; Macaca mulatta ; Torque teno virus ; genetics ; isolation & purification ; physiology ; Virus Replication

OBJECTIVETo discuss the diagnostic value of ultrasonic examination in patients with early liver cirrhosis and the relation with different stages of liver fibrosis.

METHODSIn the series, 263 patients with chronic hepatitis B were under taken liver biopsy and ultrasonic examination of type B for determination of liver cirrhosis images, width of the main portal vein and the splenic vein, tumefaction of the spleen. Data were analysed statistically.

RESULTSSixties of 263 patients were diagnosed as early liver cirrhosis. The diagnostic sensitivity, specificity, misdiagnostic rate, missed diagnostic rate, and Jonden's index of ultrasonic examination for early liver cirrhosis were 52.5%, 88.3%, 11.7%, 47.5%, and 0.508, respectively. The width of the main portal vein with liver fibrosis of S1, S2, S3, and S4 were 10.93 mm +/- 1.25 mm, 11.35 mm +/- 1.06 mm,11.29 mm +/- 1.52 mm, and 11.4 8mm +/- 1.25 mm, respectively with statistic difference between S4 and S1 (P=0.03). The width of the spleen vein of S1, S2, S3, and S4 were 6.518 mm +/- 2.033 mm, 7.190 mm +/- 1.569 mm, 7.444 mm +/- 1.805 mm and 8.406 mm +/- 2.227 mm, respectively with statistic difference between S4 and S2 (P=0.035). The incidence of tumefaction of the spleen was increased with the degree of liver fibrosis.

CONCLUSIONSThe diagnostic sensitivity of ultrasonic examination for early liver cirrhosis is low. The width of the main portal vein, the spleen vein and the incidence of tumefaction of the spleen are related with the degree of liver fibrosis. The regeneration node of liver cirrhosis may contribute to the development of portal hypertension.

Adolescent ; Adult ; Child ; Female ; Hepatitis B, Chronic ; complications ; Humans ; Liver Cirrhosis ; complications ; diagnostic imaging ; pathology ; Male ; Portal Vein ; diagnostic imaging ; pathology ; Sensitivity and Specificity ; Splenic Vein ; diagnostic imaging ; pathology ; Ultrasonography, Doppler, Color ; methods

Genetic Variation ; Hepatitis B Core Antigens ; genetics ; Hepatitis B e Antigens ; genetics ; Hepatitis B virus ; genetics ; immunology ; Humans ; T-Lymphocytes, Cytotoxic ; immunology

OBJECTIVETo establish a simple and practicable method to identify the different genotypes of hepatitis B virus (HBV).

METHODSBased on the alignment of 114 complete nucleotide sequence for HBV DNA of different genotypes, the specific sequence of each genotype was found. Six primer sets were designed for each of the six genotypes according to the genotype-specific sequence, and used separately for PCR. The genotype of HBV was identified according to the positive result of PCR. Three primer sets for B, C and D genotypes were added into a single tube for PCR reaction, and HBV was genotyped according to the length of the amplified DNA.

RESULTSThere was no difference in the genotyping result of PCR by single or multiplex primers, which was identical to the PCR-RFLP method.

CONCLUSIONSThis multiplex PCR method is simple, precise, sensitive, and easy to use.

DNA Primers ; Databases, Nucleic Acid ; Genotype ; Hepatitis B virus ; classification ; genetics ; Humans ; Polymerase Chain Reaction ; methods ; Sequence Alignment ; methods

OBJECTIVETo investigate the clinical and histological characteristics of fibrosing cholestatic hepatitis (FCH) and the therapeutic effect of lamivudine.

METHODSBy retrospective analysis, 17 cases developed severe jaundice in 794 renal-transplanted recipients, and of them, FCH was clinically suggested in 11 and confirmed by liver biopsy in 6 cases.

RESULTSThe prevalence of chronic HBV infection in renal transplantation patients was 9.3%, of whom the FCH occurred in 22.9%. In 6 liver-biopsied cases, the onset was within 1.5-22 months. Two cases remitted who had early received lamivudine and 4 cases who were treated with the drug before transplantation did not develop the disease. All patients received large amounts of multiple immuno-suppressors after transplantation. About one fifth of HBV-infected cases gradually developed cholestatic hepatitis and some of them rapidly proceeded to hepatic failure. All had very high serum level of HBV DNA. The histology revealed unique lesion combination. The hepatocytes had widespread ballooning change and some ground-glass appearance. There were liver cytolysis and focal cell loss, bile stasis, periportal fibrosis, while only mild lymphocytic infiltration.

CONCLUSIONSFibrosing cholestatic hepatitis may happen following renal transplantation. Lamivudine has marked therapeutic effect for FCH.

Adult ; Anti-HIV Agents ; therapeutic use ; Cholestasis, Intrahepatic ; drug therapy ; etiology ; pathology ; DNA, Viral ; blood ; genetics ; Female ; Fibrosis ; Hepatitis B ; blood ; complications ; Hepatitis B virus ; genetics ; Hepatocytes ; drug effects ; pathology ; Humans ; Kidney Transplantation ; adverse effects ; Lamivudine ; therapeutic use ; Male ; Middle Aged ; Retrospective Studies