Increased levels of soluble HLA‐G molecules in Tunisian patients with chronic hepatitis B infection

Hepatitis B virus (HBV) infection is a global health problem. The mechanisms of immune tolerance in HBV infection are still unclear. The host immune response plays a critical role in determining the outcome of HBV infection. Human leucocyte antigen‐G (HLA‐G) is involved in immunotolerogenic process and infectious diseases. This study aimed to explore the implication of soluble HLA‐G (sHLA‐G) and its isoforms in HBV infection. Total sHLA‐G (including shedding HLA‐G1 and HLA‐G5) was analysed by ELISA in 95 chronic HBV patients, 83 spontaneously resolvers and 100 healthy controls (HC). To explore the presence of sHLA‐G dimers, we performed an immunoprecipitation and a Western blot analysis on positive samples for sHLA‐G in ELISA. The serum levels of sHLA‐G were significantly increased in patients with chronic HBV patients compared to spontaneously resolvers and HC (P<.0001). Interestingly, we found an increased level of sHLA‐G1 in chronic HBV patients than in spontaneously resolvers and HC (P<.001). In addition, the expression of HLA‐G5 seems to be higher in the sera of chronic HBV patients than spontaneously resolvers (P=.026). The analysis of HLA‐G dimers showed the presence of homodimers in 93% of chronic HBV patients, 67% in spontaneously resolvers and 60% in HC. These results provide evidence that sHLA‐G may have a crucial role in the outcome of HBV infection and could be proposed as a biomarker for infection outcome. Based on its tolerogenic function, HLA‐G might be considered as a new promising immunotherapeutic approach to treat the chronic infection with HBV.     

Relationship between HLA‐DPA1 mRNA expression and susceptibility to hepatitis B

Chronic hepatitis B virus (HBV) infection is influenced by both viral and host factors. In genome‐wide association studies, the human leucocyte antigen HLA‐DPA1 and related polymorphism rs3077 were found to be associated with susceptibility to and spontaneous clearance of HBV infection. Here, we evaluated the association between HLA‐DPA1 mRNA expression and the risk of HBV infection. HLA‐DPA1 and rs3077 polymorphisms were investigated in 169 patients with chronic HBV and 217 healthy controls (HCs) from Sichuan Han blood donors using sequence‐based typing and meta‐analysis for HLA‐DPA1 alleles. HLA‐DPA1 mRNA levels were measured by real‐time polymerase chain reaction. The results showed that HLA‐DPA1 and rs3077 were associated with HBV infection in the Sichuan population. Rs3077T and DPA1*01:03 played protective roles in HBV infection, and rs3077C and DPA1*02:02 increased susceptibility to HBV infection. We found that the HLA‐DPA1 mRNA expression was decreased in the CHB group; in particular, the 3077CT, 3077TT, DPA1*01:03 and DPA1*02:01 alleles showed a significant decrease. Our results demonstrated, for the first time, that expression of HLA‐DPA1 alleles and rs3077 affected the risk of HBV infection. Genotypes with lower HLA‐DPA1 expression had a greater susceptibility to HBV infection. Thus, further independent studies are needed to strengthen the associations of these polymorphisms with susceptibility to and clearance of HBV infection in Chinese populations.     

Association of human leukocyte antigen polymorphism with outcomes of hepatitis B virus infection

Background and Aim: Host genetic and environmental factors are viewed as a common basis of the different outcomes of hepatitis B virus (HBV) infection. Human leukocyte antigen (HLA) plays an important role in immunological reaction to HBV infection. In this study, we aimed to determine the association between HBV infection and HLA‐A, B, and DRB1 alleles in northern Iran. Methods: HLA‐A, B, and DRB1 alleles in 33 patients with chronic hepatitis B (CHB) and 31 healthy carriers as the persistent group, and 30 subjects who had spontaneously recovered from HBV infection were analyzed by using the polymerase chain reaction (PCR)–sequence‐specific primer (PCR‐SSP) technique. Results: The frequency of the HLA‐A*33 allele was higher in the persistent group than in the recovered group (10.16% vs 0%, P < 0.008); the frequency of the DRB1*13 allele was lower in the persistent group than in the recovered group (3.13% vs 11.67%, P < 0.03). The frequency of the B*52 allele was higher in CHB patients than healthy carriers (7.58% vs 0%, P < 0.05). The logistic regression model showed that the presence of the HLA‐DRB1*13 allele was the significant factor associated with protection against the persistency of HBV. There were significant differences between the HBV recovered group, CHB patients, and healthy carriers regarding age, hepatitis B e antigen, and anti‐hepatitis B e positivity. Conclusion: HLA‐A*33 was closely related with susceptibility to persisting hepatitis B infection, and HLA‐DRB1*13 was closely related with protection against persisting hepatitis B in an Iranian population. These findings emphasized that the host HLA polymorphism is an important factor in determining the outcome of HBV infection.     

慢性乙型肝炎病毒感染者肝组织HBcAg表达与临床及病理特征的关系

目的探讨慢性HBV感染者肝组织HBcAg表达及其在亚细胞结构的分布与血清HBeAg表达、HBV DNA水平及肝组织病理学损害的关系。方法对371例慢性HBV感染者进行肝穿刺活检,检测肝组织HBcAg、血清HBV标志物、血清ALT及血清中的HBV DNA水平。比较HBcAg阴性与HBcAg表达为核型、浆型、浆核型的病例血清HBeAg阳性率、HBV DNA水平与肝组织病理学损害的关系。同时观察血清HBeAg阳性组及阴性组,肝细胞HBcAg不同表达形式在不同年龄阶段的分布特点。结果肝组织HBcAg阴性组血清HBeAg阳性率为33.1%,比HBcAg阳性各组均低(核型组68.7%;浆型组62.3%;浆核型组84.5%),炎症分级G≥2的患者比例为21.5%,低于HBcAg阳性各组(核型34.3%;浆型67.7%;浆核型69.1%),P<0.01,差异有统计学意义。其中浆型组及浆核型组G≥2的患者比例高于核型组,P<0.01,差异有统计学意义。血清HBV DNA水平在核型组高于其他各组,P<0.05,差异有统计学意义。血清HBeAg阳性组中,年龄≤20岁的HBcAg肝细胞表达为核型的比例为61.5%,高于年龄为20～39岁的11.5%及年龄≥40岁的12.3%,随年龄增加浆型及浆核型表达的比例增加,在三组分别为23.1%/7.7%、26.4%/30.8%、28.4%/45.4%,差异有统计学意义(χ2=53.74,P<0.01)。血清HBeAg阴性组中,肝组织HBcAg阴性的在各年龄组均占大部分(68.1%、61.5%、40.4%)。随年龄增加浆型及浆核型表达的比例增加,在三组分别为4.6%/4.6%、19.3%/7.7%、26.9%/20.4%,差异无统计学意义(χ2=8.94,P>0.05)。结论慢性乙型肝炎患者肝组织HBcAg表达与血清HBeAg表达有关。浆型及浆核型HBcAg表达常伴随明显的肝组织炎症。HBcAg表达形式在不同年龄阶段具有不同特点,与其所处自然史阶段有关。     

Novel hepatitis B virus surface antigen mutations associated with occult genotype B hepatitis B virus infection affect HBsAg detection

The causative factors of occult hepatitis B infection are complicated and not yet been fully elucidated. Mutations in hepatitis B virus (HBV) S gene are one of the factors may contributing to occult infection. In this study, 89 blood donors with genotype B occult HBV infection were investigated. Fifty‐seven hepatitis B surface antigen (HBsAg)‐positive/HBV DNA‐positive blood donors served as control group for comparison. Occult HBV‐related mutations with a high incidence (P < .05) in the S gene were identified. To further verify these occult infection‐related mutations, a conservative full‐gene expression vector of HBV B genotype (pHBV1.3B) was constructed. Then, the mutant plasmids on the basis of pHBV1.3B were constructed and transfected into HepG2 cells. Extracellular as well as intracellular HBsAg was analysed by electrochemical luminescence and cellular immunohistochemistry. Ten occult infection‐related mutations (E2G, Q101R, K122R, M133T, D144E, G145R, V168A, S174N, L175S and I226S) were significantly more frequent in the occult infection group (P < .05). Five of the ten mutations (E2G, D144E, G145R, V168A and S174N) strongly decreased extracellular HBsAg level (P < .05) in the transfection system. Notably, the E2G mutation had the most significant impact on the ratio of extracellular HBsAg (3.8% vs pHBV1.3B) and intracellular HBsAg (239.3% vs pHBV1.3B) (P < .05), and the fluorescence density of E2G mutant HBsAg was significantly higher than that of pHBV1.3B (P < .0001). Hence, ten mutations were associated with genotype B occult HBV infection; E2G and V168A were novel mutations which we confirmed significantly affect HBsAg detection. E2G might cause HBsAg secretion impairment that results in intracellular accumulation and a decrease in HBsAg secretion.     

Hepatic HLA antigen display in chronic hepatitis B virus infection

To determine the relationship between hepatitis B virus (HBV) replication and HLA antigen display at a cellular level, the hepatic expression of HLA antigens and HBV genome (HBV-DNA)/gene products (HB_sAg/HB_cAg) was studied in 24 patients with chronic HBV infection using simultaneously immunohistochemistry and nonisotopicin situ hybridization. The effect of interferon- and interferon- on hepatocyte HLA antigen expression was also evaluated using primary hepatocyte culture in eight patients with chronic HBV infection. HLA class I antigens were detected on hepatocyte membrane in 23 patients (95.8%). Hepatocytes positive for HB_cAg and HBV-DNA (cytoplasmic ± nuclear) were either negative or only faintly positive for HLA class I antigens, while hepatocytes positive for HB_sAg showed similar levels of HLA class I antigen expression compared with those hepatocytes with no HB_sAg expression. In contrast, hepatocytes adjacent to inflammatory infiltrates, whether positive for HBV-DNA or HBV antigens or not, were always strongly positive for HLA class I antigens. Furthermore, active liver histology (N=12) was associated with a higher overall level of hepatic HLA class I antigen expression as compared with inactive histology (N=12,P=0.003). Both interferon- and interferon- treatmentin vitro enhanced hepatocyte HLA class I antigen expression. These data indicate that expression of HLA class I antigens is not enhanced on the membrane of hepatocytes with HBV replication, and this may be one factor that permits the development of viral persistence.     

YMDD耐药变异与HLA等位基因多态性的相关性

目的：初步探讨慢性乙型肝炎(CHB)患者拉米夫定治疗中YMDD变异与HLA-A,B,DRB1各位点等位基因分布频率的相关性．方法：对142例CHB患者,采用荧光标记杂交双探针PCR融解曲线法(FH-PCR-MC)检测血浆HBV YMDD变异；对其中56例患者的外周血白细胞,采用序列特异性引物/聚合酶链式反应(PCR-SSP)技术检测人类白细胞表面抗原等位基因(HLA-A-B,DRB1)分型．结果：在用拉米夫定治疗的142例CHB患者中,YMDD变异率为56．3％．HLA-B~*58和DRB1~*03等位基因分布频率在YMDD变异组与YMDD野生组比较有显著性降低(0．013 vs 0．094,P=0．036；0．000 vs 0．063,P=0．024)；HLA-A~*30等位基因分布频率在YIDD组明显增高,与YVDD组比较差异显著(0．158 vs 0．024,P=0．034)；HLA-A~*33等位基因分布频率在YVDD变异组明显增高,与YIDD变异组比较差异显著(0．119 vs 0．000,P=0．028)．结论：YMDD耐药变异与HLA等位基因多态性有一定相关性．携有HLA-B~*58和DRB1~*03等位基因的个体感染的HBV可能不易发生YMDD变异；携有HLA-A~*30等位基因的个体感染的HBV可能易发生YIDD变异：携有HLA-A~*33等位基因的个体感染的HBV可能易发生YVDD变异．     

Correlation of hepatocyte expression of hepatitis B viral antigens with histological activity and viral titer in chronic hepatitis B virus infection: an immunohistochemical study

Background and Aim: The localization of hepatitis B virus (HBV) core antigen to the nucleus or cytoplasm of hepatocytes has biological implications for viral packaging and persistence. This study examined the relationship between the localization of hepatitis B virus antigens, histological activity, and viral titer in patients with chronic HBV infection. Methods: Liver biopsies from 110 patients with chronic HBV infection were studied. Ishak's scoring system was used for the histological analysis. The localization of hepatitis B surface antigen (HBsAg) and hepatitis B core antigen (HBcAg) and the percentage of hepatocytes stained positive by immunohistochemistry were correlated with viral titer, histological activity, and fibrosis indices using Spearman rank correlation. Results: In 88 hepatitis B e‐antigen (HBeAg)‐positive individuals, the nuclear localization of HBcAg correlated significantly with DNA titer (r = 0.435, P = 0.001) and negatively with fibrosis (r = ?0.297, P = 0.005). The cytoplasmic localization correlated significantly with histological activity (r = 0.211, P = 0.049). In 22 HBeAg‐negative individuals, the nuclear localization of HBcAg correlated significantly with histological activity (r = 0.625, P = 0.002), DNA titer (r = 0.651, P = 0.009), and fibrosis (r = 0.447, P = 0.042). The cytoplasmic localization correlated significantly with DNA titer (r = 0.524, P = 0.045) and fibrosis (r = 0.528, P = 0.012). There was no correlation of HBsAg expression with DNA titer, histological activity index, or fibrosis in both groups. HBeAg‐positive patients presented at a younger age. Conclusion: In HBeAg‐positive individuals, nuclear core antigen correlated with DNA titer, and cytoplasmic localization with histological activity, whereas in HBeAg‐negative individuals, nuclear localization correlated with DNA titer, histological activity, and fibrosis, and cytoplasmic localization correlated with DNA titer and fibrosis, but not with histological activity. These observations suggest biological differences between HBeAg‐positive and ‐negative disease.     

Long-term clinical impact of occult hepatitis B virus infection in chronic hepatitis B patients

BACKGROUND/AIMS: Long-term clinical outcomes of occult hepatitis B virus (HBV) infection were studied. METHODS: Fifteen chronic hepatitis B patients were monitored for a median of 4.4 years (range 0.9-15.3) after hepatitis B surface antigen (HBsAg) seroclearance. Serum HBV DNA was measured by real-time detection polymerase chain reaction. Thirteen patients underwent liver biopsies at the end of follow-up and liver histology was evaluated by Ishak score. Liver HBV DNA was also measured for 12 patients. RESULTS: At the end of follow-up, HBV viremia was absent in 13 (87%) patients, and antibody titers to hepatitis B core antigen showed an inverse correlation with time from HBsAg seroclearance (r=-0.554; P=0.0040). However, all patients retained liver HBV DNA and tested positive for the covalently closed circular HBV DNA replicative intermediate. The hepatic HBV DNA loads had no relation to liver histology. Paired biopsies from 11 patients disclosed that each necroinflammatory score significantly improved after HBsAg seroclearance. Amelioration of liver fibrosis was also evident in eight (73%) patients (P=0.0391 by signed rank test). CONCLUSIONS: A long-standing but strongly suppressed HBV infection may confer histological amelioration after HBsAg seroclearance.     

Hepatitis B surface antigen seroconversion is associated with favourable long-term clinical outcomes during lamivudine treatment in HBeAg-negative chronic hepatitis B patients

The aims of this study were to assess hepatitis B surface antigen (HBsAg) seroconversion and to determine its impact on the natural course of the disease in patients with HBeAg-negative chronic hepatitis B (CHB) during lamivudine (LMV) treatment. A total of 183 consecutive patients with HBeAg-negative CHB who were treated with LMV were included in the study. Data were retrospectively collected from outpatient visit charts. The primary endpoint was HBsAg seroconversion to anti-HBs. The secondary endpoint was to determine the development of cirrhosis. Loss of HBsAg was confirmed in 10 patients and seroconversion to anti-HBs in nine patients during LMV treatment or after its discontinuation. HBsAg seroconversion was achieved on-treatment in four patients after a median treatment duration of 30 months and off-treatment in the remaining five patients in a median 61 months after LMV discontinuation. The cumulative probability of HBsAg seroconversion increased from 0.6% at 1 year and 1.9% at 5 years to 21.5% at 10 years of LMV during and after LMV treatment. HBsAg clearance was preceded by undetectable serum hepatitis B virus (HBV) DNA. The majority of the patients responding to treatment had undetectable HBV DNA levels at 24 weeks of treatment. The cumulative probability of LMV resistance increased from 2.2% at 1 year to 37.3% at 5 years. No baseline parameter predicting either HBsAg seroconversion or the emergence of LMV resistance was identified. None of the patients with HBsAg seroconversion experienced virological breakthrough or disease progression during the follow-up period. These results indicate that HBsAg seroclearance can occur in patients with HBeAg-negative CHB under LMV therapy and predicts better clinical outcome.     

Mutations associated with occult hepatitis B virus infection result in decreased surface antigen expression in vitro

Summary. Occult hepatitis B virus (HBV) infection is characterized by the absence of detectable hepatitis B surface antigen (HBsAg) in the serum, despite detectable HBV DNA. Investigations of the mechanisms underlying the development of occult HBV infection are lacking in the current literature, although viral mutations in the surface region, resulting in decreased HBsAg expression or secretion, represent one potential mechanism. Wild‐type HBsAg expression vectors were constructed from genotype‐matched chronic HBV sequences. Site‐directed mutagenesis was then utilized to introduce three genotype A mutations – M103I, K122R and G145A – associated with occult HBV infection in vivo, alone and in combination, into the wild‐type HBsAg vectors. Transfection of Huh7 and HepG2 cell lines was performed, and cell culture supernatants and cell lysates were collected over 7 days to assess the effects of these mutations on extracellular and intracellular HBsAg levels. The G145A mutation resulted in significantly decreased extracellular and intracellular HBsAg expression in vitro. The most pronounced reduction in HBsAg expression was observed when all three mutations were present. The mutations evaluated in vitro in the current study resulted in decreased HBsAg expression and potentially increased hepatic retention and/or decreased hepatic secretion of synthesized HBsAg, which could explain the lack of HBsAg detection that is characteristic of occult HBV infection in vivo.     

GWAS identifying HLA‐DPB1 gene variants associated with responsiveness to hepatitis B virus vaccination in Koreans: Independent association of HLA‐DPB1*04:02 possessing rs1042169 G ‐ rs9277355 C ‐ rs9277356 A

Recently, HLA class II loci, including HLA‐DPB1, have been reported to be associated with interindividual variance in the hepatitis B (HB) vaccine response. In this study, we investigated significant single nucleotide polymorphisms (SNPs) for anti‐HBs antibody levels in 6867 healthy Koreans using a genome‐wide association study (GWAS). In GWAS, the top 20 SNPs that showed significant association with anti‐HBs levels (P < 1.0 × 10^?29) all resided in HLA‐DPB1. Utilizing PCR sequencing, we verified the relationship of the top 3 most significant SNPs (rs1042169, rs9277355 and rs9277356) from the GWAS and genotypes of HLA‐DPB1 with the HB vaccine response in Korean infants who received a scheduled vaccination. The DPB1*04:02 allele has G, C and A nucleotides for the 3SNP sites, and was significantly more frequent in responders than in nonresponders (10.9% vs 1.0%, P_c = 0.018). DPB1*05:01 was significantly more frequent in nonresponders than in responders (49.0% vs 31.1%, P_c = 0.018). In multivariate logistic regression, DPB1*04:02 showed a significant association with both vaccine response (P = 0.037, OR = 8.465) and high‐titre response (P = 0.027, OR = 9.860). The haplotypes rs1042169 G ‐ rs9277355 C ‐ rs9277356 A showed a significant association with a high‐titre response only (P = 0.002, OR = 2.941). In conclusion, DPB1*04:02 possessing rs1042169 G ‐ rs9277355 C ‐ rs9277356 A is an independent predictor of the HB vaccine response in Koreans.     

Hepatocyte hepatitis B surface antigen expression in chronic hepatitis B virus carriers in Singapore: Correlation with viral replication and liver pathology

The hepatocyte hepatitis B surface antigen (HBsAg) expression in 149 liver biopsies from 124 chronic hepatitis B virus (HBV) carriers was correlated with serum HBV DNA status and histologic activity. Hepatocyte HBsAg was stained by the peroxidase-antiperoxidase method and serum HBV DNA was determined by dot blot hybridization. Sixty-five biopsies (44%) showed minimal changes (MC), 82 biopsies (55%) showed chronic liver disease (CLD) and 2 biopsies (1%) showed hepatocellular carcinoma. Hepatocyte HBsAg was found in 144 biopsies (97%). It was present in the cytoplasm of 141 specimens (95%) and/or plasma membrane of 48 specimens (32%). Approximately half (45%) of the cytoplasmic HBsAg-positive biopsies showed discrete distribution, while the other half (55%) were grouped. Fifty-five per cent (77 of 141) of cytoplasmic HbsAg-positive biopsies had CLD, while 44% (62 of 141) showed MC. There was no relationship between the presence of cytoplasmic HBsAg or its topographic distribution with disease activity. Membrane HBsAg distribution was similar for both groups of patients (MC vs CLD: 25 of 65 (38%) vs 23 of 82 (28%); P = NS). Serum HBV DNA was detected in 98 patients (66%) and was seen mostly in association with CLD (CLD vs MC: 61% vs 39%, P less than 0.001). It was also detected more often in the sera of patients with membrane HBsAg than in those with cytoplasmic HBsAg staining (41 of 48 (85%) vs 97 of 141 (67%); P less than 0.02). However, discrete distribution of cytoplasmic HBsAg was associated with positive serum HBV DNA when compared with grouped distribution (52 of 63 (83%) vs 43 of 78 vs (55%); P less than 0.005).     

HBeAg血清转换延迟患者的临床病理特征

目的探讨HBV感染者延迟HBeAg血清转换与肝脏病理变化的相关性。方法分析148例慢性HBV感染者的血清HBeAg表达与肝脏病理改变和HBcAg的关系。结果本组病例中HBeAg阳性78例,肝脏病理炎症分级G≥2者,53例（67.9%）,肝脏病理纤维化分期S≥2者,29例（37.2%）,HBeAg阴性70例,G≥2者,36例（51.4%）,S≥2者,25例（35.7%）,HBeAg阳性与阴性组肝脏炎症损害差异有统计学意义,χ2=4.20,P〈0.05,纤维化程度差异无统计学意义,χ2=0.532,P〉0.05。HBeAg阳性组肝脏HBcAg阳性者有60例（76.9%）明显高于HBeAg阴性组26例（37.1%）,χ2=23.98,P〈0.01。两组均以胞浆型为主分别为58.3%,65.4%。结论免疫活动期,HBV感染者延迟HBeAg血清转换组与HBeAg阴性组比较有更明显的炎症程度,血清HBeAg表达与肝脏HBcAg表达有明显相关性。     

Association of an HLA‐G 14‐bp Insertion/Deletion polymorphism with high HBV replication in chronic hepatitis

Identification of an HLA‐G 14‐bp Insertion/Deletion (Ins/Del) polymorphism at the 3′ untranslated region of HLA‐G revealed its importance in HLA‐G mRNA stability and HLA‐G protein level variation. We evaluated the association between the HLA‐G 14‐bp Ins/Del polymorphism in patients with chronic Hepatitis B virus (HBV) infection in a case–control study. Genomic DNA was extracted from 263 patients with chronic HBV hepatitis and 246 control subjects and was examined for the HLA‐G 14‐bp Ins/Del polymorphism by PCR. The polymorphic variants were genotyped in chronic HBV seropositive cases stratified according to HBV DNA levels, fibrosis stages and in a control population. There was no statistical significant association between the 14‐bp Ins/Del polymorphism and increased susceptibility to HBV infection neither for alleles (P = 0.09) nor for genotypes (P = 0.18). The stratification of HBV patients based on HBV DNA levels revealed an association between the 14‐bp Ins/Del polymorphism and an enhanced HBV activity with high HBV DNA levels. In particular, the Ins allele was significantly associated with high HBV DNA levels (P = 0.0024, OR = 1.71, 95% CI 1.2–2.4). The genotype Ins/Ins was associated with a 2.5‐fold (95% CI, 1.29–4.88) increased risk of susceptibility to high HBV replication compared with the Del/Del and Ins/Del genotypes. This susceptibility is linked to the presence of two Ins alleles. No association was observed between the 14‐bp Ins/Del polymorphism and fibrosis stage of HBV infection. We observed an association between the 14‐bp Ins/Del polymorphism and high HBV replication characterized by high HBV DNA levels in chronic HBV patients. These results suggest a potential prognostic value for disease outcome evaluation.     

Inhibition of hepatitis B virus by oxymatrine in vivo

AIM To investigate the anti-HBV effect ofoxymatrine (oxy) in vivo.METHODS HBV transgenic mice were producedby micro-injection of a 4.2kb fragmentcontaining the complete HBV genomes.Expression level of HBsAg and HBcAg in thetransgenic mice liver was determined byimmunohistochemical assay,RESULTS Four groups (6 mice in each group)were injected intraperitoneally with oxy at thedosage of 100,200, and 300 mg/kg or with salineonce a day for 30 days. Both HBsAg and HBcAgwere positive in livers of all the six mice in thecontrol group (injected with saline), and werepositive in livers of two mice in 100 mg/kg groupand 300mg/kg group. In 200mg/kg group,HBsAg and HBcAg were negative in livers of allthe six mice. Based on the results, 200 mg/kg isthe ideal dosage to explore the effect of oxy atdifferent time points. According to the oxytreatment time, mice were divided into fourgroups: 10 d, 20 d, 30 d and 60 d (4 mice in eachgroup). Each mouse underwent liver biopsy twoweeks before the treatment of oxy. Down-regulation of HBsAg and HBcAg appeared aftertreatment of oxymatrine for 10 d and 20 d, Dane-like particles disappeared after the treatment ofoxy for 20d under electron microscopy,however, the expression level of HBsAg andHBcAg returned to normal 60 d later after oxytreatment.CONCLUSION oxymatrine can reduce thecontents of HBsAg and HBcAg in transgenic miceliver, longer treatment time and larger dosagedo not yield better effects.