Efficacy and safety of AnluoHuaxian pills on chronic hepatitis B with normal or minimally elevated alanine transaminase and early liver fibrosis: A randomized controlled trial.

ETHNOPHARMACOLOGICAL RELEVANCE: The AnluoHuaxian pill (AHP) is a widely used patented medicine for chronic hepatitis B (CHB) patients with advanced fibrosis or cirrhosis that has been used in China for more than 15 years. However, data are lacking on whether monotherapy with AHP can be effective in CHB patients with alanine aminotransferase (ALT) levels less than 2 times the upper limit of normal (ALT<2ULN) and early liver fibrosis (F ≤ 2). AIM OF THE STUDY: We aimed to investigate whether monotherapy with AHP improves liver histology in these patients. MATERIALS AND METHODS: In this double-blind, randomized, placebo-controlled trial, 270 CHB patients with ALT<2ULN and F ≤ 2 were treated in 12 hospitals in China. The patients were randomly assigned to an intervention (AHP) group and a placebo group at a ratio of 2:1. Of these 270 enrolled patients, 147 had paired liver biopsies. The primary end point was histological change after 48 weeks of treatment. RESULTS: Per-protocol analysis revealed that the rate of histologic improvement in liver fibrosis patients in the AHP group was significantly higher than that in the placebo group (37.7% vs. 19.5%, P = 0.035) after 48 weeks of treatment, which was consistent with results from intention-to-treat and sensitivity analyses. Moreover, after adjusting for baseline characteristics, AHP was superior to placebo with respect to improving liver fibrosis (odds ratio [OR] = 2.58, 95% confidence interval [CI]: (1.01, 6.63),P = 0.049) and liver histology (OR = 3.62, 95% CI: (1.42, 9.20),P = 0.007). In noninvasive measurement of liver fibrosis (FibroScan®), the level of liver stiffness measurement (LSM) had decreased significantly at 48 weeks (5.1 kPa) compared with that at baseline (5.7 kPa) (P = 0.008) in the AHP group, whereas it did not decrease significantly in the placebo group. Cirrhosis developed in one patient in the placebo group but in no patients in the AHP group. No serious side effects occurred in the AHP-treated patients. CONCLUSIONS: Treatment of CHB patients who had ALT<2ULN and F ≤ 2 with the traditional Chinese medicine AHP for 48 weeks improves liver fibrosis. However, due to the short duration of treatment and the limited sample size of liver pathology, the long-term benefits of AHP in reducing fibrosis and the risk of cirrhosis and hepatocellular carcinoma in these patients need to be further studied in the future.

Traditional Chinese Medicine Syndrome Patterns and Their Association with Hepatitis B Surface Antigen Levels during the Natural History of Chronic Hepatitis B Virus Infection.

The aim of this study is to investigate traditional Chinese medicine syndrome (TCMS) patterns and their association with hepatitis B surface antigen (HBsAg) levels during the natural history of chronic hepatitis B virus infection (CHB). Patients were categorized according to the phase of CHB, as follows: immune tolerance (ITP); immune clearance (ICP); low or nonreplication (LRP); reactivation (RAP); hepatic cirrhosis (HC); and primary liver cancer (PLC). TCMS patterns were classified among the following types: spleen-kidney deficiency (SKD); liver-qi depression (LQD); damp-heat in liver-gallbladder (LGDH); liver-kidney deficiency (LKD); and blood stasis blocking collateral (BSBC). HBsAg levels and other serological indicators were quantified for all patients and their association with TCMS was statistically analyzed and determined. Two hundred and eighty-nine patients with CHB were included. During the natural history of CHB, TCMS patterns were statistically different among the different phases (P < 0.001). The most frequently occurring syndromes among the six progressive phases were SKD, LGDH, LKD, LGDH, BSBC, and LGDH, respectively. The predominant patterns in the inactive stage (ITP + LRP), active stage (ICP + RAP), and late or advanced stage (HC + PLC) were SKD (31%), LGDH (51.8%) and BSBC (34.4%), respectively. Median HBsAg levels were also statistically different among the five patterns of TCMS (P < 0.001). The highest HBsAg levels were observed in SKD (4.48 log10 IU/mL). Medium levels were in LQD (3.91 log10 IU/mL) and LGDH (3.90 log10 IU/mL). The lowest HBsAg levels were in LKD (3.60 log10 IU/mL) and the second lowest levels in BSBC (3.81 log10 IU/mL). In addition, HBsAg levels in LKD and BSBC were significantly lower than those in SKD, LQD, and LGDH (P < 0.05 or 0.001). TCMS was altered during the natural history of CHB and correlated with HBsAg titers. This study could provide further insight into the therapy of CHB.

Addition and Subtraction Theory of TCM Using Xiao-Chaihu-Decoction and Naturopathy in Predicting Survival Outcomes of Primary Liver Cancer Patients: A Prospective Cohort Study.

To investigate the therapeutic effect of combined Xiao-Chaihu-Decoction and naturopathic medicine therapy on survival outcomes of patients' PLC. In XCHD group (n = 76), patients were treated with Xiao-Chaihu-Decoction in accordance with the addition and subtraction theory of TCM; in NM group (n = 89), patients were managed by naturopathic medicine; in combined group (n = 70), the same volume of Xiao-Chaihu-Decoction combined with naturopathic medicine procedures was applied. There were no evident statistical differences of age, gender, KPS score, body weight, smoking status, AFP levels, HbsAg status, TBIL levels, tumor diameters, and numbers among different groups, showing comparability among groups. No significant difference was found regarding the total remission rate and stability rate of tumors in patients treated by Xiao-Chaihu-Decoction and naturopathic medicine, except the combined therapy. KPS scores were significantly improved after treatment among groups. After treatment, 52.8% cases maintained a stable or slight increase in weight, of which 42.1%, 48.3%, and 70.0% cases maintained weight stably in the XCHD group, NM group, and combined treatment group, respectively. Xiao-Chaihu-Decoction associated with naturopathy may predict improved prognostic outcomes in PLC patients, along with improved remission and stability rates, increased KPS scores, and stable weight maintenance.

Hepatic resection versus transarterial chemoembolization for patients with Barcelona Clinic Liver Cancer intermediate stage Child-Pugh A hepatocellular carcinoma.

The present study aimed to compare the overall and recurrence-free survival rates following hepatic resection (HR) and transcatheter arterial chemoembolization (TACE) in patients with Barcelona Clinic Liver Cancer (BCLC) classified intermediate-stage Child-Pugh A hepatocellular carcinoma (HCC). A total of 443 patients were examined, among whom 274 underwent HR, whereas 169 received TACE. The overall survival, recurrence-free survival between groups and subgroups, and risk factors with respect to mortality and recurrence, were analyzed. The 1-, 3- and 5-year overall and recurrence-free survival rates were 70, 46 and 37% and 73, 52, and 37%, respectively after HR, compared with 38, 15, and 12% and 44, 25 and 16%, respectively after TACE. Overall and recurrence-free survival rates were significantly increased following HR compared with TACE. Subgroup analysis in the multi-nodule group showed that the 1-, 3- and 5-year overall survival rates were 68, 38 and 30% after HR, compared with 36, 10 and 0% following TACE. In the solitary tumor group, 1-, 3- and 5-year overall survival rates were 71, 50 and 38% after HR, and 41, 22 and 15% after TACE. The overall survival rate after HR was significantly increased compared with that after TACE in the solitary tumor and multi-nodule groups. The risk factors for mortality include solitary tumor diameter >10 cm, multi-nodules, serum albumin level ≥35 g/l, prothrombin time >13 sec, alphafetoprotein levels >400 ng/ml, and patients with hepatitis B virus. Solitary tumor diameter >10 cm, multi-nodules, and hepatitis B virus (P<0.001) were found to be associated with higher recurrence of HCC. Overall and recurrence-free survival rates were improved after HR compared with those after TACE in BCLC stage B, Child-Pugh A, HCC patients.

[Prevention and treatment mechanism of qingxia therapy (based on yinchenhao decoction and dachengqi decoction) on hepatocyte apoptosis in rats with acute hepatic injury induced by lipopolysaccharide/D-galactosamine].

OBJECTIVE: To study the prevention and treatment mechanism of Qingxia therapy (based on Yinchenhao Decoction and Dachengqi Decoction) on hepatocyte apoptosis in rats with acute hepatic injury induced by lipopolysaccharide plus D-galactosamine (LPS/D-GalN). METHODS: The acute hepatic injury model was established by LPS/D-GalN and then intervened with Qingxia therapy. Serum liver function, PT and liver tissue pathology were observed, hepatocyte apoptosis index was detected by Tunel, protein expressions of BCL-2, BAX and Caspase-3 were detected by Western blotting. RESULTS: Qingxia therapy could significantly decrease serum ALT, AST and TBIL levels (P < 0.01 or 0.05), reduce hepatocyte necrosis and inflammatory cell infiltration. There were more apoptotic cells in model group, which had significant differences compared with Qingxia group and control group. Protein expressions of BAX and Caspase-3 in model group were significantly higher than those in control group and Qingxia group (P < 0.05), but BCL-2 protein expression in model group was lower (P < 0.05). CONCLUSION: Qingxia therapy can ameliorate the liver function and hepatic tissue pathology of rats with hepatic injury induced by LPS/D-GalN, alleviate hepatocyte apoptosis in rats, prevent and treat hepatocyte apoptosis by down-regulating the protein expressions of Caspase-3 and BAX, up-regulating the protein expression of BCL-2, and adjusting the balance of BCL-2/BAX.

Chinese medicine syndrome distribution of chronic hepatitis B virus carriers in immunotolerant phase.

OBJECTIVE: To explore Chinese medicine (CM) syndrome distribution of chronic hepatitis B virus (HBV) carriers in immunotolerant phase (ITP). METHODS: One hundred and eighty-five chronic HBV carriers in ITP, seen in the Third Affiliated Hospital of Sun Yat-sen University from May 2009 to December 2010, were admitted in an observational study under the guidance of CM. Patients' CM symptoms and signs, demographics, liver biochemistries, and qualitative HBV DNA were recorded in the questionnaires. CM syndromes were then differentiated to 15 detailed types and analyzed by generalization. Lastly, the location, pathogenic factors and nature of the disease were also assessed. RESULTS: When CM syndrome patterns were differentiated to 15 types, there were 27 (15%) no syndrome cases, 94 (50%) single syndrome cases and 64 (35%) compound syndromes cases. The main detailed syndromes included Liver (Gan)-qi depression (LQD), Kidney (Shen)-qi deficiency (KQD), Spleen (Pi)-qi deficiency (SQD) and Kidney-yang deficiency (KYAD). After CM syndromes generalized to five types, their frequency was Spleen-Kidney deficiency (SKD)>LQD>inner dampness-heat retention (IDHR)>Liver-Kidney deficiency (LKD)>blood stasis blocking collateral (BSBC). SKD and LQD occupied 64%. The disease location included Liver, Gallbladder (Dan), Spleen, Stomach (Wei) and Kidney. The pathogenic factors were mainly qi stagnation, qi deficiency, yang deficiency, concurrently dampness-heat and blood stasis. The deficiency syndrome was more than excess syndrome in its nature. CONCLUSIONS: Most of chronic HBV carriers in ITP have their CM syndrome, and the most common types are SKAD, LQD. This study suggests that the natural history may be improved through breaking the state of immune tolerance or shorten the time of ITP by strengthening Spleen-Kidney and reliving Liver qi.

Down-regulation of telomerase activity and activation of caspase-3 are responsible for Tanshinone I-induced apoptosis in monocyte leukemia cells in vitro.

Tanshinone I (Tan-I) is a diterpene quinone extracted from the traditional herbal medicine Salvia miltiorrhiza Bunge. Recently, Tan-I has been reported to have anti-tumor effects. In this study, we investigated the growth inhibition and apoptosis inducing effects of Tan-I on three kinds of monocytic leukemia cells (U937, THP-1 and SHI 1). Cell viability was measured by MTT assay. Cell apoptosis was assessed by flow cytometry (FCM) and AnnexinV/PI staining. Reverse transcriptase polymerase chain reaction (RT-PCR) and PCR-enzyme-linked immunosorbent assay (ELISA) were used to detect human telomerase reverse transcriptase (hTERT) expression and telomerase activity before and after apoptosis. The activity of caspase-3 was determined by Caspase colorimetric assay kit and Western blot analysis. Expression of the anti-apoptotic gene Survivin was assayed by Western blot and Real-time RT-PCR using the ABI PRISM 7500 Sequence Detection System. The results revealed that Tan-I could inhibit the growth of these three kinds of leukemia cells and cause apoptosis in a time- and dose-dependent manner. After treatment by Tan-I for 48 h, Western blotting showed cleavage of the caspase-3 zymogen protein with the appearance of its 17-kD subunit, and a 89-kD cleavage product of poly (ADP-ribose) polymerase (PARP), a known substrate of caspase-3, was also found clearly. The expression of hTERT mRNA as well as activity of telomerase were decreased concurrently in a dose-dependent manner. Moreover, Real-time RT-PCR and Western blot revealed a significant down-regulation of Survivin. We therefore conclude that the induction of apoptosis by Tan-I in monocytic leukemia U937 THP-1 and SHI 1 cells is highly correlated with activation of caspase-3 and decreasing of hTERT mRNA expression and telomerase activity as well as down-regulation of Survivin expression. To our knowledge, this is the first report about the effects of Tan-I on monocytic leukemia cells.

Inactivation of PI3k/Akt signaling pathway and activation of caspase-3 are involved in tanshinone I-induced apoptosis in myeloid leukemia cells in vitro.

Tanshinone I (Tan I), a diterpene quinone extracted from herbal medicine Salvia miltiorrhiza Bunge, has recently been reported to have antitumor effects. As the mechanism of its proapoptotic effects on human myeloid leukemia cells has not been extensively studied, we performed an in-depth evaluation of the effects of Tan I on apoptosis in human K562 and HL-60 cells. The results revealed that Tan I could inhibit the growth of leukemia cells and cause apoptosis in a time- and dose-dependent manner. Apoptosis was observed clearly by flow cytometry and Hoechst 33258 staining, as well as DNA fragmentation analysis. After treatment by Tan I for 48 h, the percentage of disruption of mitochondrial membrane potential (Δψm) was increased in a dose-dependent manner. Western blotting analysis demonstrated the cleavage of caspase-3 zymogen protein and a dose-dependent cleavage of poly-(ADP-ribose) polymerase. Tan I-induced apoptosis was accompanied by a significant decrease in survivin and an increase in Bax. Moreover, Tan I treatment remarkably downregulated the phosphorylation of both P85/PI3K and Akt in a time-dependent manner, and the PI3K/AKT-specific inhibitor (LY294002) mimicked the apoptosis-inducing effects of Tan I. We therefore conclude that the induction of apoptosis by Tan I in these leukemia cells is mainly related to the disruption of Δψm, the upregulation of Bax expression, and the activation of caspase-3. This process is highly correlated with the inactivation of PI3K/Akt/survivin signaling pathways. The results indicate that Tan I may serve as an effective adjunctive reagent in the treatment of leukemia.

The protective effects of ambroxol on radiation lung injury and influence on production of transforming growth factor beta1 and tumor necrosis factor alpha.

The aim of this article was to investigate the effect of ambroxol on radiation lung injury and the expression of transforming growth factor beta(1) (TGF-beta(1)), as well as tumor necrosis factor alpha (TNF-alpha) in plasma. Totally, 120 patients with locally advanced lung cancer in radiotherapy were randomized into treatment and control groups. Patients in the treatment group took ambroxol orally at a dosage of 90 mg, three times per day for 3 months from the beginning of radiotherapy. The expression of TGF-beta(1) and TNF-alpha in plasma was analyzed. The clinical symptoms and lung diffusing capacity were monitored using high resolving power computed tomography. The level of TGF-beta(1) in the control group was increased (11.8 +/- 5.5 ng/ml), whereas in ambroxol-treated patients, the increase was not significant (5.6 +/- 2.6 ng/ml, P < 0.001). Radiotherapy-induced elevation of TNF-alpha levels, seen in control patients, was also abolished after treatment with ambroxol (5.1 +/- 1.0 vs. 2.4 +/- 0.8 ng/ml, P < 0.001). In the treatment group, carbon monoxide diffusion capacity was not significantly decreased at 6, 12, and 18 months post-radiotherapy, compared with the control group (P < 0.05). Ambroxol decreased the expression of TGF-beta(1) and TNF-alpha, and minimized the diminishment of lung diffusion capacity after radiotherapy.

[Effects of glycyrrhizin on the expression of hepatitis B virus and Toll like receptors 2,4 in HepG2.2.15 cells expressing low HBsAg].

OBJECTIVE: To investigate the effects of glycyrrhizin (GL) on the expression of hepatitis B virus e antigen (HBeAg), HBV DNA, Toll-like receptors 2,4 (TLR2,4) and proliferation of cells in HepG2.2.15 cell line. METHODS: Real-time PCR examined HBV DNA, ELISA examined HBsAg, HBeAg and MTT examined the proliferation of cells. FCM examined the positive percent of cells expressing TLR2,4 before and after stimulated with GL, in contrast to the blank group. RESULTS: The expression of HBsAg was low in the cell line, so e antigen was studied. The total HBeAg mean was significantly difference on the second day after stimulated (P<0.01), but only in the group with 400 microg/ml HBeAg decreased significantly in contrast to the blank group (P<0.05), the group with 800 microg/ml increased significantly in contrast to the other groups (P<0.01). The total HBV DNA mean on the third day after stimulated was significant, only the group with 50 microg/ml decreased in contrast to the blank group, but P>0.05, the other four groups increased. The intensity of TLR4 expression in the cells was significantly higher than that of the control group (P<0.05), both of total mean TLR2,4 increased significantly (P<0.01). The four groups except the group with 200 microg/ml increased significantly in no dose-dependent manner (P<0.05). GL in three goups under 200 microg/ml all could make cells proliferate, but only 200 microg/ml group had significant difference compared to the blank group (P<0.05). Both 400 and 800 microg/ml groups inhibited the growth of cells (P<0.01). The proliferation of cells were notably negative correlated with the expression of HBeAg, HBV DNA (P<0.05). CONCLUSION: The study suggestes GL could inhibit or promote HBV DNA replicating and e antigen secreting in mutative HepG2.2.15 cell line, the correlation between the proliferation of cells and the both are negative. GL could upregulate TLR2,4 in no dose-dependent manner. Influencing HBV maybe have no correlation to up regulating TLR2,4 by GL at least in vitro.

Ponicidin inhibits monocytic leukemia cell growth by induction of apoptosis.

In this study two monocytic leukemia cell lines, U937 and THP-1 cells, were used to investigate the anti-proliferation effects caused by ponicidin. Cell viability was measured by an MTT assay. Cell apoptosis was assessed by flow cytometry as well as DNA fragmentation analysis. Cell morphology was observed using an inverted microscope and Hoechst 33258 staining. RT-PCR and Western blot analysis were used to detect survivin as well as Bax and Bcl-2 expressions after the cells were treated with different concentrations of ponicidin. The results revealed that ponicidin could inhibit the growth of U937 and THP-1 cells significantly by induction of apoptosis. The suppression was in both time- and dose-dependent manner. Marked morphological changes of cell apoptosis were observed clearly after the cells were treated with ponicidin for 48 approximately 72 h. RT-PCR and Western blot analysis demonstrated that both survivin and Bcl-2 expressions were down-regulated remarkably while Bax expression remained constant before and after apoptosis occurred. We therefore conclude that ponicidin has significant anti-proliferation effects by inducing apoptosis on leukemia cells in vitro, downregulation of survivin as well as Bcl-2 expressions may be the important apoptosis inducing mechanisms. The results suggest that ponicidin may serve as potential therapeutic agent for leukemia.