[Study of disease burden of chronic hepatitis B and C patients in Shanghai based on Bronfenbrenner' s ecological systems theory: a community-based survey].

Objective: To systemically analyze family burden, quality of life of chronic hepatitis B and C patients in Shanghai and related influencing factors. Methods: A representative sample of chronic hepatitis patients (n=1 478) and their family members (n=1 478) was randomly selected through a multi-stage cluster sampling from 30 communities in 10 districts of Shanghai. One patient and one family member of each family were interviewed using different questionnaires to collect related information. Based on Bronfenbrenner' s ecological systems, psychological measurement, two-level random intercept model and multivariable structural equation model were applied to determine the effects and directions of the factors between life quality of chronic hepatitis patients and family burden. Results: The mean score of quality of life of chronic hepatitis patients in Shanghai was 78.70 ± 13.25, the score of " specific module" was highest and the score of " social function" was lowest. Additionally, the mean score of burden reported by the family members was 12.62±10.74, the score of " financial burden" was highest, and the score of " effect on family member' s health" was lowest. Multivariable structural equation model indicated that eight factors were related with life quality and family burden of patients with chronic hepatitis. Among them, HCV infection, elevated serum alanine aminotransferase level, average monthly cost for patient >3 000 yuan (RMB) and poor health of family members were the direct risk factors for the life quality of the patients as well as family burden. The factor of drinking more than once a week influenced the patients' life quality directly and family burden indirectly. On the contrary, the factors of local household registration, hospitalization and family member's indifferent attitude to hepatitis B vaccination influenced the family burden of the chronic hepatitis patients directly and the life quality of the patients indirectly. Conclusion: The findings could be used in the development of community based management and intervention of chronic hepatitis patients in Shanghai.

Case-control study of risk factors for human infection with avian influenza A(H7N9) virus in Shanghai, China, 2013.

The first human infection with avian influenza A(H7N9) virus was reported in Shanghai, China in March 2013. An additional 32 cases of human H7N9 infection were identified in the following months from March to April 2013 in Shanghai. Here we conducted a case-control study of the patients with H7N9 infection (n = 25) using controls matched by age, sex, and residence to determine risk factors for H7N9 infection. Our findings suggest that chronic disease and frequency of visiting a live poultry market (>10 times, or 1-9 times during the 2 weeks before illness onset) were likely to be significantly associated with H7N9 infection, with the odds ratios being 4.07 [95% confidence interval (CI) 1.32-12.56], 10.61 (95% CI 1.85-60.74), and 3.76 (95% CI 1.31-10.79), respectively. Effective strategies for live poultry market control should be reinforced and ongoing education of the public is warranted to promote behavioural changes that can help to eliminate direct or indirect contact with influenza A(H7N9) virus.

Metformin inhibits hepatitis B virus protein production and replication in human hepatoma cells.

Hepatitis B virus surface antigen (HBsAg) plays an important role in maintaining the tolerance and may interfere with host innate and adaptive immune responses; therefore, novel therapeutic strategies to reduce HBsAg loads in patients infected with hepatitis B virus (HBV) are emerging as an attractive but challenging issue. Metformin could regulate hepatic metabolism while the latter interacts with HBV infection. We hypothesized that metformin could affect HBsAg expression and HBV replication and may work synergistically when combined with current antivirals. In our study, a notably inhibitory effect on HBsAg production, as well as a moderate inhibition in HBV replication and HBeAg expression was observed following metformin treatment. The 50% effective concentration (EC50) for extracellular HBsAg and intracellular HBsAg in HBV-producing HepG2.2.15 cells was 2.85 mm and 2.75 mm, respectively, with a similarly selective index of about 18. When administered in combination, metformin enhanced the inhibitory effects of interferon-α2b on HBsAg expression and HBV replication and provided a complimentary role in HBsAg expression for lamivudine (LMV). This novel action of metformin derives partially from its inhibition on multiple HBV cis-acting elements. By the virtues of preferably hepatocyte distribution and safety profile, collectively, our results suggest that metformin would be potentially clinically helpful as an HBsAg production inhibitor.

The multidrug resistance of tumour cells was reversed by tetrandrine in vitro and in xenografts derived from human breast adenocarcinoma MCF-7/adr cells.

Multidrug resistance (MDR) is one of the main obstacles limiting the efficacy of chemotherapy treatment of tumours. One of the main causes of MDR is linked to the overexpression of P-glycoprotein (P-gp). This study aimed to characterise tetrandrine (Tet), a potent inhibitor of P-gp mediated MDR. Cytotoxicity was determined by the tetrazolium (MTT) assay. A MCF-7/adr cell xenograft model was established to investigate the effect of Tet on reversing MDR in vivo. Mechanistic experiments were conducted to examine the uptake, efflux and accumulation of doxorubicin (Dox) and Fura-2, and to assess lipid membrane fluidity. Tet potentiated the cytotoxicity of Dox; a 20.4-fold reversal of resistance was achieved in the presence of 2.5 micromol/l of Tet. Accumulation and efflux studies with the P-gp substrates, Dox and Fura-2, demonstrated that Tet inhibited the P-gp-mediated drug efflux. In addition, Tet lowered cell membrane fluidity in a concentration-dependent manner. In mice bearing the MDR MCF-7/adr cell xenografts, coadministration of Tet potentiated the antitumour activity of doxorubicin without a significant increase in toxicity. Tet was an extremely potent MDR modulator both in vitro and in vivo, without apparently enhancing the toxicity of the co-administered drugs. Hence, Tet holds great promise as a MDR modulator for the treatment of P-gp-mediated MDR cancers.

Screening and discovery of novel MDR modifiers from naturally occurring bisbenzylisoquinoline alkaloids.

BACKGROUND: The failure of conventional cancer chemotherapy has been linked to overexpression of a membrane associated P-glycoprotein (P-gp) that acts as an energy-dependent drug efflux pump. A promising strategy to conquer multidrug resistance (MDR) is to develop functional MDR modifiers that can inhibit the activity of P-gp. MATERIALS AND METHODS: We used MTT in combination with other in vitro drug evaluation assays to screen potential MDR modifiers from a series of naturally occurring Bisbenzylisoquinoline Alkaloids (BBIs) that were isolated from natural plants. RESULTS: Our in vitro screening assays indicated that at least six of these natural compounds (FF0019, FF0018, FF0015, FF0014, FF0011 and FF0012) showed potent activities to restore sensitivity of resistant tumor cells, such as MCF-7/adr and KBv200 cells, to many antitumor drugs including doxorubicin and vincristine. Further analyses by measurement of radioactive [3H]-Vincristine indicated that these BBIs increased intracellular drug accumulation in MDR cells, but had little effect on drug-sensitive cells. CONCLUSIONS: These results suggested that the mechanism of these compounds to reverse MDR was associated with the increase in the intracellular drug accumulation through inhibiting the activity of P-gp. Another important feature is that the in vitro cytotoxic effect of these naturally occurring BBIs themselves on tumor cells was very low. Thus, these compounds may possess great promise in being developed into novel MDR modifiers.

Chinese herbal formula XQ-9302: pilot study of its clinical and in vitro activity against human immunodeficiency virus.

OBJECTIVES: To evaluate the effectiveness of XQ-9302--a purified, precise mixture of 20 Chinese herbs--against infection with human immunodeficiency virus in vitro and in the clinic. DESIGN: In vitro cell culture assay, heavy metal content analysis, and pilot non-randomised clinical trial. SETTING: Drug rehabilitation centre and municipal surveillance centre, Shanghai, China. PATIENTS: Forty-eight patients who had various clinical histories, such as drug abuse, cancer, and infection with human immunodeficiency virus, participated in the clinical study. INTERVENTION: During the clinical trial, multiple 15-day courses of XQ-9302 10.8 g/d were given to participants. MAIN OUTCOME MEASURES: CD4 count, P24 antigen level, level of antibody against human immunodeficiency virus, number of copies per millilitre of human immunodeficiency virus in the plasma (viral load), and any side effects. RESULTS: XQ-9302 protected cultured MT4 cells from infection with human immunodeficiency virus in vitro. Clinical tests showed that the herbal formula relieved the symptoms of acquired immunodeficiency syndrome and enhanced CD4 counts in patients infected by the human immunodeficiency virus. There were no observable side effects, even after taking the drug for several months. In three patients who had acquired immunodeficiency syndrome, treatment with XQ-9302 reduced the magnitude of the viral load by more than 1 log. CONCLUSION: XQ-9302 not only improves the immune function of patients infected with the human immunodeficiency virus, but also interrupts viral replication and slows the progression of the disease without detectable side effects. In addition, the heavy metal content of XQ-9302 is well within safety levels set by the Government of China.

Circumvention of tumor multidrug resistance by a new annonaceous acetogenin: atemoyacin-B.

AIM: To explore the effect of atemoyacin-B (Ate) on overcoming multidrug resistance (MDR). METHODS: Bullatacin (Bul) was used as a positive control. Cytotoxic effects of Bul and Ate were studied with cell culture of human MDR breast adenocarcinoma cells, MCF-7/Dox and human KBv200 cells, and their parental sensitive cell lines MCF-7 and KB. Cytotoxicity was determined by tetrazolium (MTT) assay. The function of P-glycoprotein (P-gp) was examined by Fura 2-AM assay. Cellular accumulation of doxorubicin (Dox) was determined by fluorescence spectrophotometry. Apoptosis was measured by flow cytometry. RESULTS: IC50 of Ate for MCF-7/Dox, MCF-7, KBv200, and KB cells were 122, 120, 1.34, and 1.27 nmol.L-1, respectively. IC50 of Bul for MCF-7/Dox, MCF-7, KBv200, and KB cells were 0.60, 0.59, 0.04, and 0.04 nmol.L-1, respectively. The cytotoxicities of Bul and Ate to MDR cells were similar to those to parental sensitive cells. Bul and Ate markedly increased cellular Fura-2 and Dox accumulation in MCF-7/Dox cells, but not in MCF-7 cells. The rates of apoptosis in MDR cells were similar to those in sensitive cells induced by Ate. CONCLUSION: There was no cross-resistance of P-gp positive MCF-7/Dox and KBv200 cell lines to Bul and Ate as compared with their sensitive P-gp negative MCF-7 and KB cell lines. The mechanism of the circumvention of MDR was associated with the decrease of P-gp function and the increase of cellular drug accumulation in MDR cells.

Reversal of tumor multidrug resistance by 2-phenyl-3-(3',5'-dimorpholinomethyl-4'-hydroxy)-benzoyl-indole (HWL-12).

AIM: To explore the reversal of multidrug resistance (MDR) by indole derivative HWL-12. METHODS: Cytotoxicity was determined by tetrazolium (MTT) assay. The function of P-gp was examined by Fura 2-AM assay. Cellular accumulation of doxorubicin (Dox) was measured by fluorescence spectrophotometry. RESULTS: HWL-12 10 mumol.L-1 markedly increased Fura-2 accumulation and was 17.2-fold reversal of MDR in MCF-7/ADR cells. The cellular Dox accumulation in MDR cells was increased in the presence of HWL-12 on the MCF-7/ADR cells. No effect was observed for Dox accumulation in the presence of high Ca2+ (addition of CaCl2) or low Ca2+ (addition of egtazic acid). CONCLUSION: HWL-12 has a potent MDR reversal action which was associated with the increase of cellular Dox accumulation in MDR cells and not related with calcium ion concentration.

Effect of mitoxantrone on DNA polymerase of Ehrlich ascites carcinoma cells.

AIM: To study the effect of mitoxantrone (Mit) on DNA polymerases of tumor cells. METHODS: DNA polymerases of Ehrlich ascites carcinoma cells were isolated by phosphocellulose column chromatography. The effects of Mit on DNA polymerase alpha, beta, and gamma were detected by method of K Ono. RESULTS: Mit inhibited DNA polymerase alpha, beta, and gamma, IC50 values were 11.9, 6.5, and 11.9 mumol.L-1, and Ki 1.86, 2.22, and 2.05 mumol.L-1, respectively. The inhibitory mode of Mit on DNA polymerase alpha, beta, and gamma was competitive. CONCLUSION: Mit is a strong inhibitor on DNA polymerase alpha, beta, and gamma. The inhibitory mode was competition with respect to template DNA.

[Compared study on Fura-2/AM assay and MTT assay for screening multidrug resistant modulators].

To explore the difference of screening results of reversing multidrug resistance (MDR) by modulators between Fura-2/AM assay and MTT assay, 25 compounds which have active structure were studied for MDR reversal activity with both methods. The fold of MDR reversal was shown to have remarkable relation with the amount of Fura-2 accumulation (Y = -3.66 + 17.5X, gamma = 0.86, P < 0.01). On the other hand, Fura-2/AM assay has several advantages as compared with MTT assay. Fura-2/AM assay needs shorter time (4 h) than MTT assay (96 h), and the MTT assay needs more steps than the Fura-2/AM assay. Furthermore, Fura-2/AM assay was more reliable than MTT assay for screening MDR modulators because MTT assay was dependent on the viable cells, while Fura-2/AM assay was dependent on the function of P-gp. The results suggest that Fura-2/AM assay may replace MTT assay in the screening of MDR modulators on a large scale.

[A comparative study on effect of two bisbenzylisoquinolines, tetrandrine and berbamine, on reversal of multidrug resistance].

A comparative study on the effect of two bisbenzylisoquinolines, tetrandrine (TTD) and berbamine (BBM), and verapamil (VRP) on reversing multidrug resistance was reported. TTD, BBM and VRP showed significant activity in reversing adriamycin (ADR) and vincristine (VCR) resistance in acquired resistant MCF-7/Adr and KBv200 cell lines, and the effect was shown to be dose-dependent. TTD, at the concentration of 10 mumol.L-1, completely reversed ADR resistance in MCF-7/adr cells. TTD, BBM and VRP increased intracellular ADR accumulation in MCF-7/adr cells. There is minor difference in structure between TTD and BBM. TTD showed greater activity than VRP in reversing MDR, while BBM showed similar activity to that of VRP. TTD also showed significant activity in vivo in reversing ADR resistance in MDR MCF-7/Adr solid tumor in nude mice.

An epidemic of hepatitis A attributable to the ingestion of raw clams in Shanghai, China.

An epidemic of hepatitis A in 1988 in Shanghai had an overall attack rate of 4083/100,000 population (292,301 cases). The epidemic curve showed three peaks in January and February. A case-control study of 1208 matched pairs supported that clams were the vehicle for the virus (summary odds ratio, 9.47; P less than .001). Analysis of subsets who had eaten clams indicated that only 3.5% with hepatitis A had cooked their clams compared with 18.1% without hepatitis A, and those with the disease consumed more clams. A historical cohort study indicated that approximately 31.7% of the population had eaten clams one or more times between 9 December 1987 and 3 January 1988. The estimated attack rates in those who had and had not eaten clams were 11.93% and 0.52%, respectively (relative risk, 22.94; attributable risk, 11.41%). The three peaks in the consumption curve correlated with those in the epidemic curve. Hepatitis A virus was demonstrated in clams taken from the Shanghai markets and from the catching area.

Dehydroandrographolide succinic acid monoester as an inhibitor against the human immunodeficiency virus.

Dehydroandrographolide succinic acid monoester (DASM) is the dehydroandrographolyl ester of succinic acid; and andrographolide, from which DASM is made, is the major diterpenoid lactone found in the Chinese medicinal herb, Andrographis paniculata. DASM has been found to be an inhibitor against the human immunodeficiency virus (HIV) in vitro. It was nontoxic to the H9 cell at the concentrations of 50-200 (average, 108) micrograms/ml and was inhibitory to the HIV-1 (IIIB) at the minimal concentration of 1.6-3.1 (average 2.0) micrograms/ml. It was also inhibitory to two other strains of HIV-1 and a strain of HIV-2. This inhibitory effect could also be demonstrated in cultures of activated human blood mononuclear cells; the 50% toxic dose and the 50% HIV inhibitory dose were about 200-greater than or equal to 400 and 0.8-2 micrograms/ml, respectively. At the subtoxic concentration, DASM partially interfered with HIV-induced cell fusion and with the binding of HIV to the H9 cell. Presumably, it also interfered with HIV replication at another unidentified step(s).

The potentiation of the antitumor activity but not toxicity of bleomycin by 3-aminobenzamide.

Our previous studies have demonstrated that 3-aminobenzamide (3AB), an inhibitor of adenosine diphosphate-ribosyl transferase (ADPRT) could enhance the cytotoxicity (in vitro) and antitumor activity (in vivo) of bleomycin (BLM) A5 and peplomycin (PEP) against S-180, hepatoma and Ehrlich ascites carcinoma (EAC). In this study, it was shown that the inhibition rates (INR's) of S-180 in two experiments were increased from 42.5 and 46.1% to 66.2 and 75.9% when BLM 2.5 mg/kg/day x 8 was combined with 3AB 385.4 mg/kg/day x 8, while the decrease of body weight could not be enhanced. BLM at a dose of 5 mg/kg/day x 8 gave INR's of 64.8 and 75%, similar to the combined group but decreased the body weight more significantly. However, the addition of 3AB 385.4 mg/kg/day to BLM did not increase the acute toxicity of BLM alone. There was no significant difference of change of the body weight and subacute toxicity between the BLM and BLM + 3AB group. There was no difference of peripheral blood white cell count and the pathomorphological and ultrastructural change, wet weight and hydroxyproline content (to reflect the collagen content) of the lung of the mice between BLM alone and BLM + 3AB group. Therefore, the study provided experimental evidences for the reasonable use of nontoxic ADPRT inhibitors in adjunct to the chemotherapy of BLM in cancers.

Potentiation of the antitumor activity of cisplatin in mice by 3-aminobenzamide and nicotinamide.

3-Aminobenzamide (3AB) and nicotinamide (NA), inhibitors of adenosine-ribose transferase (ADPRT), potentiated the antitumor activity of cisplatin (DDP) on Ehrlich ascites carcinoma in mice. The mean survival times of the mice increased from 21.2-37.0 days in DDP-treated groups to 47.0-54.6 days in mice treated with DDP plus NA or 3AB. These drugs also potentiated DDP antitumor activity on sarcoma 180, with the inhibition rates increasing from 12.4%-20.8% in groups treated daily with DDP to 29.8%-46.4% in those treated with DDP plus NA or 3AB; however, neither 3AB nor NA alone showed any antitumor activity. The single-dose lethality of DDP on mice was partially reversed by either NA or 3AB. The pathological study revealed that the morphologic changes in the proximal tubules 1 month after a single dose of DDP (10 mg/kg) were partially prevented by a single protective dose (5 mmol/kg) of NA or 3AB. Our results suggest that the combination of DDP with ADPRT inhibitors might be used clinically in the future.