Abnormal expression of miR-let-7b in primary biliary cirrhosis and its clinical significance / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To evaluate the expression of microRNA (miR)-let-7b in peripheral blood cells of patients with primary biliary cirrhosis (PBC) and investigate its relationship to clinical disease parameters.</p><p><b>METHODS</b>Peripheral blood and serum samples were obtained for study from 60 PBC patients and 60 healthy controls. Peripheral blood cells were extracted and subjected to real-time PCR to measure miR-let-7b expression. Serum levels of interleukin (IL)-18, total bilirubin (TBIL), alkaline phosphatase (ALP), and gamma-glutamyl transferase (GGT) were measured by standard biochemical assays. The relationship between miR-let-7b expression and disease parameters was assessed by Spearman's rank correlation test.</p><p><b>RESULTS</b>PBC patients showed significantly lower expression of miR-let-7b in peripheral blood cells than healthy controls (P less than 0.001); moreover, the miR-let-7b expression level decreased in parallel to increases in disease severity (stage I > II / III > IV). In PBC patients, the miR-let-7b expression was significantly correlated with Mayo risk scores (r = -0.4930, P less than 0.001), IL-18 (r = -0.4643, P less than 0.001) and ALP (r = -4119, P less than 0.001), but not with TBIL or GGT.</p><p><b>CONCLUSION</b>Decreased expression of miR-let-7b may be associated with development and progression of PBC, and this miRNA may represent a novel target of improved diagnostic and preventive strategies for PBC.</p>

Establishment of a mouse model of primary biliary cirrhosis by AMA M2 autoantigen injection / 中华肝脏病杂志

<p><b>OBJECTIVES</b>To establish a primary biliary cirrhosis (PBC) model by AMAM2 autoantigen injection into C57BL/6 mice.</p><p><b>METHODS</b>Mice of the model group were immunized intraperitonealy with 200 microl of purified recombinant AMAM2 autoantigen in complete Freund's adjuvant (CFA). Mice immunized with bovine serum albumin and CFA in the same way were used as negative controls. Sixty-six weeks later, mice were sacrificed and their sera were collected. Sera samples were assayed for AMAM2 autoantibody, alkaline phosphatase (ALP), ALT and total bilirubin (TBil). Their liver, stomach, muscle and kidney tissues were sectioned and stained using HE to observe the pathological changes.</p><p><b>RESULTS</b>Antibodies to AMAM2 autoantigen were readily induced in the model group. The mice in the model group had no significant changes in the level of serum ALT and TBil but had an obvious increase of ALP (P<0.05). The stomach, muscle and kidney tissues showed no evident damage while the livers had obvious pathological changes, including bile duct degeneration or proliferation, and mononuclear cell infiltration.</p><p><b>CONCLUSION</b>The AMAM2 autoantigen-induced PBC animal model was successfully established in C57BL/6 mice in our experiment and its characteristic biochemical and pathology are quite similar to that in the early stage of human PBC. This model may provide a useful experimental approach for further study of the pathogenesis and clinical treatment of human PBC.</p>

Analysis of HLA alleles polymorphism in Chinese patients with primary biliary cirrhosis / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the frequencies of human leuckocyte antigens (HLA) -A, B and DRB1 alleles in Chinese patients with primary biliary cirrhosis (PBC) using polymerase chain reaction-based techniques, and to assess the correlation of HLA molecules with other clinical and laboratory profiles.</p><p><b>METHODS</b>Genotyping of HLA-A, B, and DRB1 were performed in 65 well-characterized patients with primary biliary cirrhosis and 431 healthy controls with PCR amplification with sequence-specific primers (PCR-SSP).</p><p><b>RESULTS</b>The frequency of DRB1*0701 was increased to 29.2% compared with 13.9% in the controls (PC < 0.05, OR = 2.55, 95% CI: 1.4 approximately 4.6). No association was found with HLA-DRB1*08 which had been constantly reported. The A*2 allele (53.8%) was more frequent in the PBC patient group but without a significant statistical difference. The frequencies for the other A, B and DRB1 alleles were similar between patients and healthy controls. There was no difference between patients with or without DRB1*0701 in some clinical and laboratory profiles.</p><p><b>CONCLUSION</b>Susceptibility to primary biliary cirrhosis in Chinese is associated with DRB1*0701 allele and differs from people in North America, South America, North Europe and even in Japan, but the association is not restricted to any particular subgroup of patients. Valine at position 78 of HLA DRbeta1 may play an important role in the pathogenesis of primary biliary cirrhosis.</p>

A study on the relationship between interleukin-10 promoter polymorphism and autoimmune liver disease / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate whether three biallelic polymorphisms at the position -592, -819 and -1082 in the promoter region of the IL-10 gene were associated with the incidence of autoimmune liver disease.</p><p><b>METHODS</b>The IL-10 -592 and IL-10-1082 polymorphisms were genotyped by polymerase chain reaction-restriction fragment length polymorphisms analysis (PCR-RFLP), while polymerase chain reaction- sequence specific primer (PCR-SSP) assay was used to detect IL-10 -819 polymorphisms.</p><p><b>RESULTS</b>Among 54 Chinese patients with AIH or 77 Chinese patients with PBC versus healthy controls, the frequency of AA, GA genotypes at IL-10 gene promoter -1082 position was 87.0% or 83.1% versus 90.0%, 13.0% or 16.9% versus 10.0%, respectively (P > 0.05), the GG genotype in Chinese populations is absent; the frequency of CC, CT, TT genotypes at IL-10 gene promoter -819 position was 11.11% or 9.1% versus 8.1%, 44.4% or 53.3% versus 45.0%, 44.4% or 37.7% versus 46.9%, respectively (P > 0.05); the frequency of CC, CA, AA genotypes at IL-10 gene promoter -592 position was 4.9% or 14.3% versus 10.0%, 51.2% or 53.3% versus 51.9%, 43.9% or 32.5% versus 38.1%, respectively (P > 0.05). No alleles differed significantly in each groups.</p><p><b>CONCLUSION</b>There were no association between IL-10 gene polymorphisms and autoimmune liver disease</p>

Genetic association of tumor necrosis factor (TNF)-alpha polymorphisms with primary biliary cirrhosis and autoimmune liver diseases in a Chinese population / 中华肝脏病杂志

<p><b>OBJECTIVE</b>Autoimmune hepatitis (AIH) and primary biliary cirrhosis (PBC) are two autoimmune diseases of unknown etiology. Genetic factors appear to be involved in the pathogenesis of both diseases. Tumor necrosis factor (TNF)-alpha is one of the proinflammatory cytokines and immunomodulators, and is implicated in the pathogenesis of AIH and PBC. In this study, we studied the association between Chinese patients with AIH, PBC and the polymorphisms in promoter-region polymorphisms of the TNF-alpha gene at position -308 and -238.</p><p><b>METHODS</b>We have investigated four candidate gene loci in 49 patients with AIH, 58 patients with PBC, and 160 healthy controls. The polymorphisms were assessed by the PCR specifically for the single-nucleotide polymorphisms.</p><p><b>RESULTS</b>We found the difference in the TNF-alpha gene at position -308 genotype distributions between Chinese health controls and Caucasian health controls. Although the percent of TNF-alpha*2 was decrease on PBC patient group (10.34% vs. 16.88%), there was no significant difference between PBC patients and health control in the Chinese. There were also no significant differences between AIH and health control on the TNF-alpha gene at position -308 and -238.</p><p><b>CONCLUSION</b>Our findings suggest that the TNF-alpha promoter-region polymorphisms distribution is different between differe of ethnic groups; there are no genetic links of the TNF-alpha promoter-region polymorphisms to AIH and PBC in Chinese.</p>

Genetic association between interleukins gene polymorphisms with primary biliary cirrhosis in Chinese population / 中国医学科学院学报

<p><b>OBJECTIVE</b>To determine the relationship between polymorphisms in the genes encoding IL-1, IL-6, and IL-10 with primary biliary cirrhosis (PBC) in Chinese population.</p><p><b>METHODS</b>Whole-blood samples were taken from 77 patients with PBC and 160 healthy controls. DNA was extracted and the polymorphisms at positions IL-1 +3953, IL-1RN intron 2, IL-6 -174, and IL-10 -1082, -819, and -592 were determined by using sequence-specific polymerase chain reaction (SSP) or polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP).</p><p><b>RESULTS</b>The frequency of IL-1RN1,1 allele in PBC group was significantly higher than in control group (90.9% vs 79.4%, P=0.026), and the frequency of IL-1RN1,2 in PBC group was significantly lower than in control group (6.5% vs 18.8%, P=0.013). There was no significant difference in the frequence of IL-1RN*2 allele between PBC group and control group (P=0.06). Of the 77 patients with PBC, 4 patients were IL-6 -174GC, 73 were IL-6 174GG. All the 160 health controls are IL-6 -174GG (P=0.0036). The frequence of IL-6 -174C allele in PBC group was significantly higher than that in control group (P=0.0038). No significant differences of polymorphisms for IL-1 +3953 and IL-10 (-1082, -819 and -592) were found between PBC group and control group.</p><p><b>CONCLUSION</b>The polymorphisms of IL-1RN and IL-6 -174G/C appear to be associated with PBC, and the polymorphisms of IL-1 +3953 and IL-10 promoter gene are not associated with PBC in a Chinese population.</p>

Prediction and identification of autoepitopes of PDC-E2 specific CD8+ CTL in primary biliary cirrhosis patients / 中国医学科学院学报

<p><b>OBJECTIVE</b>To identify autoepitopes of E2 subunit of pyruvate dehydrogenase complex (PDC-E2) specific CD8+ CTL in primary biliary cirrhosis (PBC) patients.</p><p><b>METHODS</b>An online database SYFPEITHI was applied to predict HLA-A*0201 restricted epitopes which located in PDC-E2 30-50 aa and 150-190 aa where B-cell epitopes clustered with CD4+ T-cell epitopes. T2 cell line reconstitution and stabilization assay, induction of specific CTL lines from peripheral blood mononuclear cells (PBMCs) of patients with PBC and cytotoxicity of peptides-induced CTL were performed to screen the epitopes from those candidates.</p><p><b>RESULTS</b>Five potential epitopes were predicted by database. Of the 5 candidates, two peptides 159-167 aa and 165-174 aa, with highly binding activity to HLA-A*0201 molecules, could stimulate PBMCs from most HLA-A*0201 positive PBC patients to proliferate and peptide-induced CTL lines showed specific cytotoxicity.</p><p><b>CONCLUSION</b>Peptides of KLSEGDLLA (159-167 aa) and LLAEIETDKA (165-174 aa) in the inner lipoyl domain of PDC-E2 are HLA-A*0201 restricted CD8+ CTL immunodominant epitopes in PBC.</p>

Study on the relationship of CTLA-4 -318, +49 polymorphisms with autoimmune hepatitis and primary biliary cirrhosis in a Chinese population / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the association between Chinese patients with autoimmune hepatitis (AIH), primary biliary cirrhosis (PBC) and the polymorphisms of cytotoxic T lymphocyte -associated antigen-4 (CTLA-4) gene promoter (-318) and exon 1 (+49).</p><p><b>METHODS</b>The CTLA-4 promoter (-318 T/C) and exon 1 (+49A/G) polymorphisms were genotyped via restriction fragment length polymorphism methods in 62 Chinese AIH patients, 77 Chinese PBC patients and 160 healthy controls.</p><p><b>RESULTS</b>There was no difference in the distribution of CTLA-4 promoter -318 T/C polymorphisms between AIH patients and controls, but the C allele frequency was significantly increased in patients with AIH, compared to controls (P=0.02, OR=2.43). The distribution of CTLA-4 gene exon 1 49 A/G genotypes exhibited significant difference between PBC patients and controls (P=0.006), and the frequency of G allele showed a significant increase in PBC group as compared with controls (P=0.0046, OR=1.8). Although the genotype distribution of the CTLA-4 exon 1-promoter gene displayed no significant difference between AIH and PBC patients and controls, the occurrence of GG-CC was increased in the patients of the two groups (AIH: 32.3%, PBC: 37.7%; control: 22.5%).</p><p><b>CONCLUSION</b>The above findings suggest that the polymorphisms of CTLA-4 gene probably confer susceptibility to AIH and PBC in the Chinese population.</p>

The relationship between Chlamydia pneumoniae infection and primary biliary cirrhosis / 中华肝脏病杂志

<p><b>OBJECTIVE</b>The aim of this study was to evaluate the association between Chlamydia pneumoniae (CP) infection and primary biliary cirrhosis (PBC).</p><p><b>METHODS</b>Chlamydia pneumoniae IgG and IgM were determined by enzyme-linked immunosorbent assay (ELISA) in 41 well-established PBC patients and two race-matched control groups, PHC, n = 70; healthy controls, HC, n =57).</p><p><b>RESULTS</b>The mean levels and seroprevalence of CP IgG in PBC group and PHC group were significantly higher than in the HC [(46.8 +/- 43.4) RU/ml, (49.5 +/- 45.2) RU/ml vs (28.3 +/- 32.7) RU/ml, P = 0.042 and P < 0.001 respectively; 68.3%, 71.4% vs 42.1%, chi2 values were 5.389 and 11.110 respectively]. There was a markedly elevated seroprevalence of CP IgM in patients with PBC (22.0%) compared with the PHC and HC groups. The odds ratios (ORs) for the presence of CP IgG and IgM for the PBC patients versus the HC were 2.7 (95% CI 0.9 to 6.1) and 5.1 (95% CI 1.4 to 18.5). Though there was no correlation in the level of CP IgG with total IgG in sera of patients with PBC (r=-0.857, p=0.344), CP IgM was related with the abnormally high concentrations of total IgM in the PBC group.</p><p><b>CONCLUSIONS</b>The results of this study do not support the hypothesis that infection with Chlamydia pneumoniae may be a triggering agent for PBC, but suggest that Chlamydia pneumoniae infection probably contributes to the high level of IgM presented in most of the patients with PBC</p>

Detection of anti-M2 autoantibody using recombinant BCOADC-E2 / 中华风湿病学杂志

Objective To detect anti-M_2 autoantibody using recombinant BCOADC-E_2.Methods We purified recombinant BCOADC-E_2 by Ni~(2+)affinity chromatography column and then detect anti-M_2 autoan- tibody in the sera of patients with primary biliary cirrhosis(PBC)by Western blot test(WBT)and enzyme linked immunosorbent assays(ELISA).Results Among 60 sera from PBC patients,33 were positive,all of controls were negative.Conclusion The recombinant BCOADC-E_2 can be used to detect anti-M_2 autoanti- body specifically and sensitively.It is helpful for the diagnosis of PBC.

Detection of the expression level of Toll-like receptor3 by establishing real-time fluorescence quantitative method / 中华检验医学杂志

Objective To establish a specific fluorescence quantitative method for determining the mRNA expression of Toll-like receptor3(TLR3)in peripheral blood mononuclear cells(PBMCs).Methods Using the Beacon Designer 2.1 software,specific primers and Taqman-MGB probe were designed.The plasmid pMD18-T-TLR3 was constructed as calibrator and the amplified fragment was obtained by reverse- transcript-PCR(RT-PCR).RNA quantification based on cycle threshold values(Ct)was used to establish the standard curve.According to which,the TLR3 mRNA levels in 30 normal individuals,20 patients with primary biliary cirrhosis(PBC)and 20 ones with chronic liver cirrhosis induced by HBV were calculated automatically by software after the fluorescence of PCR product was detected continuously during amplification.Results The linear detection range of the assay for TLR3 gene and ?-actin was 10~2-10~8(r= -0.9974)and 10~3～10~8(r=-0.9984),respectively.The coefficient of variation of both intra-and inter- assay reproducibility for high concentration sample were 6.7% and 8.7%,respectively,and those for low concentration sample were 12.3% and 14.0%.The TLR3 mRNA expression level ranges from 3.46?10~2- 4.51?10~3 copies/?g RNA,4.92?10~2-1.42?10~4 copies/?g RNA and 2.58?10~2-7.17?10~3 copies/?g RNA for 30 healthy individuals,20 PBC patients and 20 ones with chronic liver cirrhosis induced by HBV, respectively.Conclusion We have successfully set up a FQ-RT-PCR method for detecting TLR3 mRNA, which may be used as an excellent tool for the clinic and basic study on the expression of TLR3 gene.

Effects of oxidized low density lipoprotein and antioxidant vitamin E on the exp ression of CD40 and CD40 ligand in cultured human monocytes / 第二军医大学学报

Objective： To investigate the effect of oxLDL and VitE on the expression of CD40 and CD40 ligand(CD40L) in cultured human monoc ytes. Methods： The expression of CD40 and CD40L on monocytes su rface were measured by indirect immunorescence technique in combination with flo w cytometry. Results： Low concentration of oxLDL（≤200 μg/L） significantly increased the expression of CD40 and CD40L in a dose and time dep endent manner. High concentration (>200 μg/L）of oxLDL markedly reduced the exp ression of CD40 and CD40L. When VitE was added, it significantly reduced the ex pression of CD40 and CD40L on monocytes surface induced by oxLDL in a dose-depe ndent manner. Conclusion：It is an important mechanism that the high expression of CD40 and CD40L induced by oxLDL may be contributed to the for mation of atherosclerosis. Antioxidan VitE can partially inhibit the high expres sion of CD40 and CD40L on monocytes surface induced by oxLDL.

Effects of cytokines on the expression of CD40 and CD40 ligand in cultured human monocytes/macrophages / 第二军医大学学报

Objective： To investigate the effect of c ytokines (IFN-γ，TNF and IL-1) on the expression of CD40 and CD40 ligand (CD4 0L) in monocytes/macrophages. Methods： The mRNA expression of C D40 and CD40L was measured by RT-PCR and the CD40，CD40L expression on the mono cytes/macrophages were detected by flow cytometric analysis. Results： IFN-γ，TNF and IL-1 could not only significantly up-regulate the mRNA levels of CD40 and CD40L in cultured monocytes/macrophages, but also increase t he expression of CD40 and CD40L. Antioxidant VitE could reduce the expression o f CD40 and CD40L induced by IFN-γ，TNF and IL-1. Conclusion： IFN-γ，TNF and IL-1 can stimulate high expression of CD40 and CD40L . Antio xidant VitE can partially inhibit the expression of CD40 and CD40L induced by cy tokines in cultured monocytes/macrophages.

Effects of hyaluronidase and hyaluronan on proliferation of va scular endothelial cells and the expression of CD44 / 第二军医大学学报

Objective: To investigate the effects of hyaluroni dase (HAase) and hyaluronan (HA) on proliferation of vascular endothelial cells and its mechanism. Methods: The cultured bovine aortic endothel ial cells (BAEC) were treated with HAase or HA. Cell proliferation rate was dete cted by MTT assay. The expression of CD44 and DNA content of the cells were meas ured by flow cytometry (FCM). Results: HAase (50 μg/ml) stimula ted cell proliferation ［(50.10±1.23）% vs control, P<0.01］, incre ased S phase cell rate and induced the expression of CD44, but HA (100 μg/ml) i nhibited cell proliferation and the expression of CD44. Conclusion: HAase may degrade antiangiogenic HA of extracellular matrix, which may stim ulate proliferation of endothelial cells and enhance the curative effect of grow th factors to myocardial ischemia.

Effect of probucol on adhesion of human monocytic line THP-1 induced by oxidiz ed low density lipoprotein / 第二军医大学学报

Objective: To determine the eff ect of probucol on adhesion of human monocytic line THP-1 induced by oxidized low density lipoprotein (oxLDL). Methods: THP-1 cells were induced by oxLDL in vitro. The CD11b, CD54 expressions and adhesion to human umbilic al vein endothelial cells (HUVEC) were measured after treatment with probucol at different concentrations by flow cytometry and β-nitrophenyl N-acetyl-β-D -glucosminide test. Results: Probucol inhibited the adhesion of oxLDL-induced THP-1 cells to HUVEC and down regulated the expression of CD11b in a dose dependent manner (P<0.01), but there was no inhibition on exp ression of CD54. Conclusion: Probucol can inhibit adhesion and a ggregation of monocyte-macrophages to endothelium in circulation, and may have anti-inflammatory action.

Effects of oxidized low density lipoprotein and antioxidant vitamin E on the exp ression of CD40 and CD40 ligand in cultured human monocytes / 第二军医大学学报

Objective： To investigate the effect of oxLDL and VitE on the expression of CD40 and CD40 ligand(CD40L) in cultured human monoc ytes. Methods： The expression of CD40 and CD40L on monocytes su rface were measured by indirect immunorescence technique in combination with flo w cytometry. Results： Low concentration of oxLDL（≤200 μg/L） significantly increased the expression of CD40 and CD40L in a dose and time dep endent manner. High concentration (>200 μg/L）of oxLDL markedly reduced the exp ression of CD40 and CD40L. When VitE was added, it significantly reduced the ex pression of CD40 and CD40L on monocytes surface induced by oxLDL in a dose-depe ndent manner. Conclusion：It is an important mechanism that the high expression of CD40 and CD40L induced by oxLDL may be contributed to the for mation of atherosclerosis. Antioxidan VitE can partially inhibit the high expres sion of CD40 and CD40L on monocytes surface induced by oxLDL.

Effects of cytokines on the expression of CD40 and CD40 ligand in cultured human monocytes/macrophages / 第二军医大学学报

Objective： To investigate the effect of c ytokines (IFN-γ，TNF and IL-1) on the expression of CD40 and CD40 ligand (CD4 0L) in monocytes/macrophages. Methods： The mRNA expression of C D40 and CD40L was measured by RT-PCR and the CD40，CD40L expression on the mono cytes/macrophages were detected by flow cytometric analysis. Results： IFN-γ，TNF and IL-1 could not only significantly up-regulate the mRNA levels of CD40 and CD40L in cultured monocytes/macrophages, but also increase t he expression of CD40 and CD40L. Antioxidant VitE could reduce the expression o f CD40 and CD40L induced by IFN-γ，TNF and IL-1. Conclusion： IFN-γ，TNF and IL-1 can stimulate high expression of CD40 and CD40L . Antio xidant VitE can partially inhibit the expression of CD40 and CD40L induced by cy tokines in cultured monocytes/macrophages.

Effects of hyaluronidase and hyaluronan on proliferation of va scular endothelial cells and the expression of CD44 / 第二军医大学学报

Objective: To investigate the effects of hyaluroni dase (HAase) and hyaluronan (HA) on proliferation of vascular endothelial cells and its mechanism. Methods: The cultured bovine aortic endothel ial cells (BAEC) were treated with HAase or HA. Cell proliferation rate was dete cted by MTT assay. The expression of CD44 and DNA content of the cells were meas ured by flow cytometry (FCM). Results: HAase (50 μg/ml) stimula ted cell proliferation ［(50.10±1.23）% vs control, P<0.01］, incre ased S phase cell rate and induced the expression of CD44, but HA (100 μg/ml) i nhibited cell proliferation and the expression of CD44. Conclusion: HAase may degrade antiangiogenic HA of extracellular matrix, which may stim ulate proliferation of endothelial cells and enhance the curative effect of grow th factors to myocardial ischemia.

Effect of probucol on adhesion of human monocytic line THP-1 induced by oxidiz ed low density lipoprotein / 第二军医大学学报

Objective: To determine the eff ect of probucol on adhesion of human monocytic line THP-1 induced by oxidized low density lipoprotein (oxLDL). Methods: THP-1 cells were induced by oxLDL in vitro. The CD11b, CD54 expressions and adhesion to human umbilic al vein endothelial cells (HUVEC) were measured after treatment with probucol at different concentrations by flow cytometry and β-nitrophenyl N-acetyl-β-D -glucosminide test. Results: Probucol inhibited the adhesion of oxLDL-induced THP-1 cells to HUVEC and down regulated the expression of CD11b in a dose dependent manner (P<0.01), but there was no inhibition on exp ression of CD54. Conclusion: Probucol can inhibit adhesion and a ggregation of monocyte-macrophages to endothelium in circulation, and may have anti-inflammatory action.

Correlation analysis between HBV Pre-S1 antigen with HBeAg and HBV DNA / 第二军医大学学报

Objective:To study the correlation between HBV Pre-S1 antigen,HBeAg levels and HBV DNA copies,so as to assess the clinical value of Pre-S1 in detection of HBV replication.Methods:A total of 363 HBsAg-positive samples were col- lected.The levels of Pre-S1 antigen,HBeAg and HBV DNA copies were determined by ELISA,time-resolved immuno-fluores- cent method and real-time fluorescent quantitative PCR(FQ-PCR),respectively.The correlation between the determination re- sults was analyzed.Results:Pre-S1 antigen level was correlated with the level of HBeAg(X~2=94.4,P