3,4,5-Tri-O-caffeoylquinic acid methyl ester isolated from Lonicera japonica Thunb. Flower buds facilitates hepatitis B virus replication in HepG2.2.15 cells.

Caffeoylquinic acids are well known for their prominent antiviral activities. Beyond our expectations, we initially found 3,4,5-Tri-O-caffeoylquinic acid methyl ester (3,4,5-CQME) from L. japonica can facilitate HBV DNA and antigens secretion. This study aimed to investigate its underlying molecular mechanism. The results indicate that 3,4,5-CQME signally increased intracellular and secreted HBsAg levels by more than two times in HepG2.2.15 cells and HepAD38 cells. Furthermore, levels of HBeAg, HBV DNA and RNA were significantly enhanced by 3-day 3,4,5-CQME treatment; it didn't directly affect intracellular cccDNA amount, although it slightly increased cccDNA accumulation as a HBV DNA replication feedback. In addition, treatment with 3,4,5-CQME significantly induced HBx protein expression for viral replication. We utilized a phospho-antibody assay to profile the signal transduction change by 3,4,5-CQME to illuminate its molecular mechanism. The results indicate that treatment with 3,4,5-CQME activated AKT/mTOR, MAPK and NF-κB pathways verified by immunoblot. Moreover, 3,4,5-CQME upregulated the expression of nuclear transcriptional factors PGC1α and PPARα. In short, 3,4,5-CQME promotes HBV transcription and replication by upregulating HBx expression and activating HBV transcriptional regulation-related signals. As caffeoylquinic acids are widely present in traditional Chinese medicines, the risk of intaking caffeoylquinic acids-containing herbs for hepatitis B treatment requires more evaluation and further research.

ANTIVIRAL ACTIVITY OF DIANTHUS SUPERBUSN L. AGAINST HEPATITIS B VIRUS IN VITRO AND IN VIVO.

BACKGROUND: Hepatitis is a viral infection of hepatitis B virus (HBV). Limitations of drug used in the management of it opens the interest related to alternative medicine. The given study deals with the antiviral activity of Dianthus superbusn L. (DSL) against HBV in vitro & in vivo. MATERIAL AND METHODS: In vitro study liver cell line HepG2.2.15 was used by transinfected it with HBV. Cytotoxicity stduy was performed by using different concentrations of DSL such as 50, 100, 200, 500 & 1000 µg/ml. Anti HBV activity of DSL was estimated by assesing the concentration of HBsAg and HbeAg in cell culture medium by using ELISA. Whereas in vivo study was performed on ducklings and antiviral activity of DSL (100, 200, 400 mg/kg) was confirmed by estimating the serum concentration of HBV DNA and histopathology study of hepatocytes in HBV infected ducklings. RESULT: Result of the study suggested that >500 µg/ml concentration of hydroalcoholic extract of DSL was found tobe cytotoxic. It was also observed that DSL significantly (p<0.05) reduces the concentration of antigenes in cell culture media as per the concentration and days of treatment dependent. Moreover in vivo study confirms the anti viral activity of DSL (200 & 400 mg/kg) as it significantly (p<0.05) decreases the serum concenetration of HBV DNA in HBV infected dukling compared to control group. Histopathology study was also reveals the hepatprotective effect of DSL in HBV infected ducklings. CONCLUSION: The given study concludes the antiviral activity DSL against HBV by in vitro and in vivo models.

¿Es posible suspender el tratamiento con antivirales orales en los pacientes con hepatitis crónica B antígeno e negativa? Experiencia y nuevas expectativas / Is it possible to stop treatment with nucleos(t)ide analogs in patients with e-antigen negative chronic hepatitis B? Experience and new expectations

INTRODUCCIÓN: El tratamiento de la hepatitis crónica B antígeno e negativa (HCB HBeAg negativa) con antivíricos orales (AO) suele prolongarse de forma indefinida debido a que la pérdida del antígeno de superficie como objetivo para su suspensión es un hecho infrecuente. Recientemente han aparecido las primeras evidencias que sugieren finalizar la terapia con AO en casos seleccionados. OBJETIVOS: Analizar la tasa de rebote virológico en pacientes con HCB Age negativa que suspendieron el tratamiento con AO. MATERIAL Y MÉTODOS: Estudio retrospectivo observacional que incluyó 140 casos de HCB HBeAg negativa. Veintidós pacientes, que recibieron exclusivamente AO, los suspendieron por diversos motivos realizándose un seguimiento posterior. Todos presentaban transaminasas normales, ADN indetectable y ausencia de cirrosis o comorbilidades importantes al finalizar el tratamiento. RESULTADOS: Doce pacientes presentaron rebote virológico (54,54%), transcurriendo una media de 6,38 meses (± 1,9) desde la suspensión hasta el rebote (el 75% dentro de los 12 primeros meses tras la suspensión). Cinco recibieron adefovir, uno lamivudina más adefovir, uno tenofovir y 5 lamivudina. La duración media del tratamiento, desde el inicio hasta la suspensión, fue de 38,5 meses (± 4,5). El grupo con respuesta sostenida presentaba una edad media y duración del tratamiento superior a los sujetos con rebote, si bien estas diferencias no resultaron estadísticamente significativas. CONCLUSIONES: Los resultados sugieren que es posible suspender la terapia con AO en casos seleccionados de HCB Age negativa, siempre que no exista cirrosis, se cumpla un tiempo mínimo de tratamiento, las transaminasas sean normales y el ADN indetectable de forma mantenida. En estos casos, se debe realizar un seguimiento estrecho durante el primer año y posteriormente de forma indefinida

BACKGROUND: Treatment of HBeAg-negative chronic hepatitis B (CHB) with nucleos(t)ide analogues (NA) is usually indefinite, since the loss of HBsAg, as a criterion for its discontinuation, is a rare event. Recent evidence suggests that discontinuing NA therapy may be feasible in selected patients. OBJECTIVES: To analyze the rate of virological relapse in patients with HBeAg-negative CHB who discontinued treatment with NAs. METHODS: We performed a single-center observational study that included 140 patients with HBsAg-negative CHB. Twenty-two patients, who received only NAs, discontinued treatment for different reasons and were subsequently monitored. All had normal ALT and AST, undetectable DNA and absence of cirrhosis or significant comorbidities before stopping treatment. RESULTS: Twelve patients showed virologic relapse (54.54%). The mean interval between discontinuation and relapse was 6.38 months (± 1.9) (75% relapsed during the first 12 months after discontinuation). Five received adefovir, 1 lamivudine and adefovir, 1 tenofovir and 5 lamivudine alone. The mean treatment duration in this group was 38.5 months (± 4.5). The sustained response group had a higher mean age and longer treatment duration than patients with virologic relapse but these differences were not statistically significant. CONCLUSIONS: The results suggest that NA treatment can be stopped in selected patients with CHB as long as they are not cirrhotic, have completed a minimum period of treatment, have normal ALT and sustained undetectable DNA. These patients should be closely monitored during the first year and then indefinitely

Predictors of relapse in chronic hepatitis B after discontinuation of anti-viral therapy.

BACKGROUND: Optimal duration of anti-viral therapy in chronic hepatitis B virus (HBV) infection remains unclear. AIM: To investigate factors that could predict relapse after stopping anti-viral agents. METHODS: Chronic hepatitis B patients who were treated with anti-viral agents (lamivudine, adefovir, entecavir) and have stopped the treatment were recruited. Anti-viral agents were stopped according to the recommendations of the Asian Pacific Association for the Study of the Liver. Virological relapse was defined as an increase in serum HBV DNA to >1000 copies/mL after discontinuation of treatment. RESULTS: Eighty-four (69 treatment naïve and 15 lamivudine resistant) patients were eligible for this study. Thirty-seven patients developed virological relapse at 4.3 ± 2.9 (range 1-11) months after discontinuation of therapy. The 1-year cumulative probability of virological relapse was 42% and 47% in HBeAg (hepatitis B e antigen)-positive (n = 41) and HBeAg (hepatitis B e antigen)-negative (n = 43) patients, respectively. On multivariate analysis by Cox proportional hazard model, pre-existing lamivudine resistance, delayed suppression of HBV DNA to undetectable level during anti-viral therapy and to a higher HBsAg (hepatitis B surface antigen) level at the end of treatment were associated with virological relapse. Twelve of the 15 (80%) lamivudine resistant patients developed virological relapse. Among the 11 treatment naïve patients who had HBsAg ≤ 2 log IU/mL at the end of treatment, 1 (9%) of them had virological relapse. CONCLUSIONS: Treatment cessation among lamivudine resistant patients is associated with high risk of virological relapse. Serum HBsAg level at the end of treatment and rate of HBV DNA suppression can provide supplementary information to guide the timing of stopping anti-viral drugs.

El virus de la hepatitis B en niños recién inmigrados / Hepatitis B serology in immigrant children

Fundamento. La inmigración es importante en nuestro medio. Algunos protocolos de atención al niño inmigrante incluyen la determinación del estado serológico respecto al virus de la hepatitis B (VHB). Objetivo. Valorar la conveniencia de la determinación sistemática del antígeno de superficie (HBsAg), anticuerpo contra el core (HBSerologia del virus de l’hepatitis B en infants recentment immigrats Ana Estabanell 1, Rosa M. Masvidal 2, Elisa de Frutos 2, Dolors Riera 3, Cecilia Cruz 1, Beatriz Miguel 2 1 CAP Gòtic (annex Rull). ABS Gòtic. Institut Català de la Salut. Barcelona. 2 CAP Dr. Lluís Sayé. ABS Raval Nord. Institut Català de la Salut. Barcelona. 3 CAP Drassanes. ABS Raval Sud. Institut Municipal d’Assistència Sanitària de Barcelona. cAc), anticuerpo contra el Ag de superficie (HBsAc), en niños inmigrantes. Determinar la prevalencia del HBsAc positivo en los vacunados contra el VHB. Método. Se determinaron HbsAg, HBsAc, HBcAc en niños de 6 meses a 15 años procedentes de países de baja renta que habían llegado hacía menos de 12 meses. Se registró su estado vacunal. Se estimó la prevalencia y el intervalo de confianza del resultado. Resultados. De los 1.226 niños/as, se determinó el HbsAg en 1.098 (89,5%), siendo 8 positivos: 0,79% (intervalo de confianza (IC) 95%: 0,37-1,43), dos de éstos constaban como vacunados para el VHB. En 1.024 se determinó el HBcAc, siendo el único marcador positivo en 7 casos: 0,98% (IC 0,47-1,80); el HBsAc se determinó en 987 y fue positivo en el 33,23%. De los 333 niños vacunados con tres dosis, en 267 se determinó el HBsAc, siendo positivo en el 59,9% (IC 95%: 55,15-64,45). Conclusiones. Consideramos indicada la determinación del HbsAg a los niños inmigrantes, incluyendo los que aportan datos de vacunación del VHB completa. No creemos justificado solicitar el HBcAc de manera sistemática. En niños inmigrantes vacunados puede estar indicado determinar los HBsAc(AU)

Background. Immigration is an important phenomenon in our environment. Some guidelines for the care of the immigrant child include the evaluation of hepatitis B virus (HBV) serology. Objective. To evaluate the need for the systematic measurement of surface antigen (HBsAg), core antigen antibody (HBcAb), and surface antigen antibody (HBsAb) in young immigrants, and to determine the prevalence of HBsAb in those who receive HBV vaccination. Method. HBsAg, HBsAb, and HBcAb values were determined in children from 6 months to 15 years of age who emigrated from low-income countries within the prior 12 months. The prevalence and confidence intervals were calculated. Results. Of the 1,226 children evaluated, HBsAg was measured in 1,098 (89.5%) and was positive in 8 (0.79%, CI 0.37-1.43), including two cases in whom vaccination had been documented; HBcAb was measured in 1,024 and was the only positive HBV marker in 7 cases (0.98%, CI 0.47-1.80); and HBsAb was measured in 987 and was positive in 33.26%. In 30.8% of the children, the only positive marker was HBsAb. Of the 333 children Background. Immigration is an important phenomenon in our environment. Some guidelines for the care of the immigrant child include the evaluation of hepatitis B virus (HBV) serology. Objective. To evaluate the need for the systematic measurement of surface antigen (HBsAg), core antigen antibody (HBcAb), and surface antigen antibody (HBsAb) in young immigrants, and to determine the prevalence of HBsAb in those who receive HBV vaccination. Method. HBsAg, HBsAb, and HBcAb values were determined in children from 6 months to 15 years of age who emigrated from low-income countries within the prior 12 months. The prevalence and confidence intervals were calculated. Results. Of the 1,226 children evaluated, HBsAg was measured in 1,098 (89.5%) and was positive in 8 (0.79%, CI 0.37-1.43), including two cases in whom vaccination had been documented; HBcAb was measured in 1,024 and was the only positive HBV marker in 7 cases (0.98%, CI 0.47-1.80); and HBsAb was measured in 987 and was positive in 33.26%. In 30.8% of the children, the only positive marker was HBsAb. Of the 333 children(AU)

Anti-hepatitis B virus activities of cinobufacini and its active components bufalin and cinobufagin in HepG2.2.15 cells.

Cinobufacini (Huachansu) is a Chinese medicine prepared from the skin of Bufo bufo gargarizans Cantor (Bufonidae), which has long been used in traditional Chinese medicine (TCM). The aim of present study was to examine the anti-hepatitis B virus (HBV) activities of cinobufacini and its active components bufalin and cinobufagin in the human HBV-transfected cell line HepG2.2.15. The hepatitis B surface antigen (HBsAg), hepatitis B e antigen (HBeAg), and hepatitis B core-related antigen (HBcrAg) concentrations in cell culture medium were determined by chemiluminescent enzyme immunoassay after HepG2.2.15 cells were respectively treated with different concentrations of cinobufacini, bufalin, and cinobufagin for 3 or 6 d. HBV DNA and mRNA were determined using transcription-mediated amplification and real-time polymerase chain reaction (PCR), respectively. On d 3, cinobufacini at a concentration of 1 µg/ml had no activity against HBV virological markers. However, on d 6, cinobufacini at 1 µg/ml effectively inhibited the secretion of HBsAg, HBeAg, and HBcrAg by 29.58, 32.87, and 42.52%. It was more potent than the positive control lamivudine (100 µg/ml). Bufalin and cinobufagin slightly inhibited HBV antigen secretion. Treatment with cinobufacini, bufalin, or cinobufagin had no anti-HBV effect on DNA in cell culture medium. Consistent with the HBV antigen reduction, HBV mRNA expression was markedly inhibited in comparison to the control when HepG2.2.15 cells were treated with cinobufacini, bufalin, or cinobufagin. Results suggested that cinobufacini had more potent activity against HBV antigen secretion than its components bufalin and cinobufagin and this inhibitory role was attributed to the specific inhibition of HBV mRNA expression.

[Preventive and therapeutic effects of qiongyugao on hepatocellular carcinoma via inhibition of the expression of HBxAg in hepatic carcinoma cells].

AIM: To investigate the effects of qiongyugao on the expression of hepatitis B x antigen (HBxAg) in BALB/c-nu mice into which human hepatic carcinoma cells were transplanted, and to analyze its specific mechanism in prophylaxis and treatment of hepatocellular carcinoma. METHODS: A nude mouse model with the transplantation of human hepatic carcinoma cells was established to observe the preventive and therapeutic effects of qiongyugao on the body weight and tumor weight of the mice. The expression of HBxAg in tumor and liver tissue wsa detected by immunohistochemical studies. RESULTS: Compared with model control group, prophylaxis and treatment with qiongyugao increased the body weight, depressed the tumor weight and inhibited HBxAg expression. The same efficacy was showed in both qiongyugao prophylaxis group and cyclophosphamide treatment group. CONCLUSION: Qiongyugao can slow down the growth of tumor and inhibit the expression of HBxAg, which may be an essential mechanism in prophylaxis and treatment of hepatocellular carcinoma.

Estudio comparativo de la capacidad inactivante de distintas colectas de la planta Phyllanthus orbicularis / Comparative study of the inactivating capacity of different harvests of Phyllanthus orbicularis

Se estudió la inactivación del AgsHB por extractos de 2 colectas de Phyllanthus orbicularis realizadas en diferentes épocas del año (Marzo/94 y Octubre/94) en 34 sueros positivos a este marcador, y se compararon estos resultados con los obtenidos para la colecta Agosto/93, con el objetivo de conocer la influencia de la época de colecta en la presencia de la actividad anti- hepatitis B de la planta. Todas las concentraciones ensayadas (100, 50 y 25 mg/mL) disminuyeron los valores de absorbancia en los sueros, y se logró una seroconversión a negativos estadísticamente significativa, con excepción de lo ocurrido para las concentraciones de 25 y 12,5 mg/mL de la colecta Octubre/94. La cantidad de sueros negativos fue proporcional a las concentraciones utilizadas. Se observaron diferencias estadísticas entre las colectas Agosto/93 y Octubre/94. Estos resultados sugirieron que podían ocurrir alteraciones en la presencia o concentración de los posibles componentes activos de P.orbicularis producto de la influencia de factores externos como lo son las condiciones climáticas

Estudio comparativo de la capacidad inactivante de distintas colectas de la planta Phyllanthus orbicularis / Comparative study of the inactivating capacity of different harvests of Phyllanthus orbicularis

Se estudió la inactivación del AgsHB por extractos de 2 colectas de Phyllanthus orbicularis realizadas en diferentes épocas del año (Marzo/94 y Octubre/94) en 34 sueros positivos a este marcador, y se compararon estos resultados con los obtenidos para la colecta Agosto/93, con el objetivo de conocer la influencia de la época de colecta en la presencia de la actividad anti- hepatitis B de la planta. Todas las concentraciones ensayadas (100, 50 y 25 mg/mL) disminuyeron los valores de absorbancia en los sueros, y se logró una seroconversión a negativos estadísticamente significativa, con excepción de lo ocurrido para las concentraciones de 25 y 12,5 mg/mL de la colecta Octubre/94. La cantidad de sueros negativos fue proporcional a las concentraciones utilizadas. Se observaron diferencias estadísticas entre las colectas Agosto/93 y Octubre/94. Estos resultados sugirieron que podían ocurrir alteraciones en la presencia o concentración de los posibles componentes activos de P.orbicularis producto de la influencia de factores externos como lo son las condiciones climáticas

Essential fatty acid supplementation in chronic hepatitis B.

BACKGROUND: Dietary supplementation with essential fatty acids and polyunsaturated lecithin may improve biochemical and histological parameters in liver disease. METHODS: Ten patients with serological and histological evidence of chronic hepatitis B received capsules of the polyunsaturated fatty acid-rich evening primrose oil in a dose of 4 g daily for 12 months, while a matched group received liquid paraffin capsules as a placebo. RESULTS: Compared to the placebo group, the patients receiving evening primrose oil showed no improvement in either biochemical or histological indices of liver damage, or in the rate of loss of circulating e antigen. CONCLUSIONS: Dietary, supplementation with this dose of essential fatty acids is unlikely to be of benefit in chronic hepatitis B.