Objective To study the characteristics of lamivudine-resistant mutation associated chronic severe hepatitis B during lamivudine treatment.Methods Twenty-seven patients with lamivudine-resistant mutation associated chronic severe hepatitis B during lamivudine treatment were analyzed retrospectively.YMDD motif mutation was detected by gene chips or DNA sequencing.The pathological features of liver tissues from 8 patients undergoing liver transplantation were analyzed.The X2 test were used to perform the stafistical analysis.Results The YMDD motif mutations of 27 lamivudine-resistant patients were 5 cases of YVDD mutation,2 of YVDD+L180M,13 of YIDD mutation,4 of YIDD+L180M,1 of YVDD+YIDD mutations,2 of YVDD+YIDD+L180M,and there was no single L180 M mutation among patients.Twenty-seven patients were divided into cirrhotic group and noncirrhotic group according to whether they were diagnosed with cirrhosis before treatment.Compared to cirrhotic group,incidence of severe hepatitis was lower,prognosis was better,the age of patients was younger and hepatitis Be antigen(HBeAg)positive rate was higher in noncirrhotic group.There were two types of pathological features of liver tissues from 8 patients,which were active hepatic cirrhosis and massive or submassive hepatic necrosis with liver shrinking significantly.Conclusions Hepatic cirrhosis is a risk factor of lamivudine-resistant mutation associated chronic severe hepatitis B.There may be two mechanisms in lamivudine-resistant mutation associated chronic severe hepatitis B.

Adolescent ; Adult ; Child ; Child, Preschool ; Hepatitis B, Chronic ; diagnosis ; therapy ; Humans ; United Kingdom

Objective To construct an X-gene defect and IFN gene contained HBV expression plasmid and explore its expression effect in HepG2 cell line. Methods By employing pBR322-B-HBV plasmid which contained type adrⅠHBV DNA, we destroyed HBV X-gene by inserting mutation and connected the defect HBV with IFN sequence. After recombination, it was introduced into a mammalian plasmid and was constructed as the objective plasmid. At the same time, two control groups were constructed. One was pcDNA3-ES-HBV2, which contained the full length of HBV DNA. The other was pcDNA3-KN-F1F2, which contained the X-gene defect HBV DNA. Then we transfected HepG2 cells with all the plasmid. After the cells were screened by G418, the X-gene defect and IFN gene contained HBV(KN-F1F2-IFN) and IFN protein from culture medium were detected by fluorescence- quantitative-PCR and ELISA respectively. Results pcDNA3-KN-F1F2-IFN, as the objective plasmid, was constructed successfully; contrary with the control virus, the amounts of both HBV and HBsAg were lower; both IFN mRNA and IFN protein were detected.Conclusions An X-gene defect and IFN gene contained HBV expression plasmid was constructed successfully and it can be expressed in HepG2 cell line.

Objective To analyze clinical courses and rescue therapies of adefovir-resistant chronic hepatitis B patients who had lamivudine resistance before and then changed to take adefovir dipivoxil. Methods 15 patients resistant to lamivudine were retrospectively analyzed, who had virological breakthrough after adefovir dipivoxil monotherapy and were treated with rescue therapy.Adefovir-resistant mutations were detected by direct sequencing of the HBV polymerase gene. Results 15 patients with former lamivudine resistance were treated with adefovir dipivoxil monotherapy for a median of 16 months, and 14 patients were found adefovir-resistant mutations at rtA181T/V and(or) rtN236T, only 1 patient was found multi-mutations at rtM204I + rtL180M + rtA181T. Rescue therapies were given to all the 15 patients after drug resistance. Among the 7 patients treated with lamivudine in combination with adefovir for 3 months,whose HBV DNA levels decreased (2.2±0.6)lg copy/mL on average, 5 patients achieved HBV DNA undetectable after 6 months combinative therapy. The HBV DNA levels of the 3 patients treated with entecavir decreased 2.8～3.5 lg copy/mL within 6 months treatment. Conclusion These preliminary data suggest the combination of lamivudine and adeforvir dipivoxil may be an effective rescue therapy for adefovir-resistant patients who have former lamivudine resistance.

OBJECTIVE: To study the therapeutic effect of Bushen Rougan Recipe (BSRGR) on hepatic fibrosis in rats. METHODS: Forty male Wistar rats were randomly divided into normal control group (n=10), model group (n=15), and BSRGR-treated group (n=15). Rats in the model and BSRGR-treated groups were administered intraperitoneally with 0.5% dimethylnitrosamine (DMN), 10 mg.kg(-1).d(-1), successive 3 days per week for 4 weeks to induce hepatic fibrosis. Then rats in the above 2 groups were given normal saline and BSRGR (10 ml.kg(-1).d(-1), ig) for another 4 weeks, respectively. Rats in the 3 groups were all executed at the end of the 8th week. The serum total bilirubin (TBIL), alanine aminotransferase (ALT), aspartate aminotransferase (AST), albumin (A) and globulin (G) were determined and the serum hyaluronic acid (HA), laminin (LN) and collagen IV (IV-C) were measured. RESULTS: The rat model of liver fibrosis was successfully induced by DMN. It was found that the serum TBIL, AST and ALT and the liver fibrosis marks were declined in BSRGR-treated group as compared with those in the model group (P<0.01). The content of total serum protein and the A/G in BSRGR-treated group were both increased as compared with those in the model group (P<0.05). CONCLUSION: BSRGR can be used to treat hepatic fibrosis in rats.

OBJECTIVE: To study the expression of TIMP-1 and TGF-beta1 mRNA in hepatic fibrosis rats and the therapeutic effects of Bushen Rougan Recipe (BSRGR). METHODS: Hepatic fibrosis was induced in rats by dimethylnitrosamine (DMN). The rats' hepatic tissue was studied by HE staining and Sirius red staining. The rats were divided into normal control group, fibrosis model group and BSRGR treated group (10 g.kg(-1).d(-1), i.g. for 4 weeks). At last, TIMP-1 and TGF-beta1 mRNA were detected by RT-PCR. RESULTS: Pathological study showed that hepatic fibrosis was successfully induced by DMN in rats. The expression quantity of TIMP-1 and TGF-beta1 mRNA were the most in the fibrosis model group, the second in the BSRGR treated group, and the least in the normal control group. CONCLUSION: The expression of TIMP-1 and TGF-beta1 mRNA was increased in the hepatic fibrosis rats, and this is one possible mechanism of hepatic fibrosis. BSRGR can inhibit the advancement of hepatic fibrosis.

To study the effect of heparin on the ConA induced acute liver injury inKunming mice,twenty four mice were randomly divided into groups A,B and C.To the mice in group A, 0 2ml of normal saline (NS) was intravenously administered, while those in group B were given ConA 18mg/kg instead of NS in order to induce severe acute liver injury. Heparin was injected subcutaneously in a dose of 100U per animal at the same time as ConA challenge in group C. Compared with group B, prior injection of heparin in group C significantly decreased the mice death rate and the peak levels of serum ALT within 8h. At the same time, intrasinusoidal congestion and hepatic inflammation were alleviated significantly,and MDA level in liver homogenate was lowered markedly. It suggested that heparin is efficient in protecting the mice from liver injury induced by ConA.

Objective To analyze the clinical features and mutation patterns of HBV polymerase gene in patients with chronic hepatitis B(CHB) after the emergence of drug-resistance during Lamivudine(LAM) therapy.Methods LAM-resistant mutations were detected by direct sequencing of the HBV polymerase gene in hospitalized patients and outpatients of CHB with LAM-resistance in Changhai Hospital from Dec.2005 to Dec.2007.Clinical features after the emergence of LAM-resistant mutations were retrospectively analyzed.Results Two hundred and fifteen patients with CHB were diagnosed as LAM-resistant.Among them 192 patients were found to have LAM-resistant-associated mutations in the HBV polymerase gene.The mean value of serum HBV DNA was 6.25?1.31(log10copies/ml),the mean value of alanine aminotransferase(ALT) was 75U/L(ranged 19-821 U/L).ALT elevation and hepatitis recrudescence were found in 139 among 192(72.4%) patients.99.0%(190/192) patients had YMDD mutations.Four major mutation patterns of LAM-resistant HBV were identified as rtM204I(33.9%),rtL180M+rtM204V(26.0%),rtL180M+rtM204I(21.9%) and rtV173L+rtL180M+rtM204V(11.5%).The rtM204V mutation was accompanied more frequently by the rtL180M mutation compared with the rtM204I mutation(P0.05).Conclusions YMDD is the major mutation pattern of HBV polymerase gene after emergence of LAM-resistance.The mutation patterns of HBV polymerase gene are possibly not related to the clinical severity of CHB patients during LAM therapy.

Objective To evaluate the efficacy and safety of adefovir dipivoxil(ADV) monotherapy and ADV lamivudine(LAM) combined therapy for patients with LAM-resistant chronic hepatitis B.Methods 124 chronic hepatitis B patients with LAM-resistant mutations were enrolled in the present study.74 patients were treated with ADV combined with LAM therapy,and other 50 patients subjected to ADV monotherapy.There were no differences between the two groups in patients' baseline characteristics.Sequencing of the HBV polymerase gene was performed to determine LAM and ADV mutations occurred at baseline or during therapy.All patients were monitored with clinical examinations and routine laboratory tests during the therapy.Results The reduced logarithmic values of serum HBV DNA after 12-week and 24-week treatment were 1.99?0.64 and 2.61?0.80 in ADV group,obviously lower compared with those in ADV+LAM group(2.55?0.74 and 3.19?0.82,respectively,P