[A phase II, single-arm, open-label, multicenter clinical study to evaluate the efficacy and safety of sofosbuvir combined with ribavirin in patients with genotype 2 chronic hepatitis C virus infection].

Objective: To evaluate the efficacy and safety of sofosbuvir (Nanjing Zhengda Tianqing Pharmaceutical Co., Ltd.) combined with ribavirin in patients with genotype 2 chronic hepatitis C virus infection. Methods: Treatment-naïve or treatment experienced genotype 2 chronic hepatitis C patients from sixteen research centers of China were screened. All subjects received once-daily dose of sofosbuvir (400 mg) combined with ribavirin (body weight < 75 kg, 1 000 mg/day, 400 mg in the morning and 600 mg in the evening; body weight > 75 kg, 1 200 mg/d, 600 mg in the morning and 600 mg in the evening) for 12 weeks. Patients were followed-up for a period of 12 weeks after discontinuation of treatment. Continuous variables were expressed as mean ± standard deviation. The proportion of subjects with virologic response at different follow-up time points and 95% confidence intervals were estimated by maximum likelihood ratio and Clopper-Pearson interval. Results: 132 cases with genotype 2 chronic hepatitis C virus infection from sixteen research centers of China were included, 12 cases of whom were associated with cirrhosis, and the remaining 120 cases were not associated with cirrhosis. One hundred and thirty-one cases completed the study, and one patient lost to follow-up at week 4 after the end of treatment. The sustained virological response rate was 96.2% (95% confidence interval: 92.37% - 99.16%) after 12 weeks of drug withdrawal. Virological relapse occurred in four cases. Of the 132 subjects enrolled in the study, 119 (90.2%) reported 617 adverse events during treatment, of which 359 (76.5%) were TEAE related to sofosbuvir and/or ribavirin. There were nine TEAEs of grade 3 and above, and six cases (4.5%) of them had six severe adverse events. Only one serious adverse event was associated with sofosbuvir and ribavirin (unstable angina pectoris). There were no adverse events leading to drug discontinuation or death. Conclusion: Sofosbuvir combined with ribavirin has a high SVR rate in the treatment of genotype 2 chronic hepatitis C virus infection, and most of the adverse events occurred were mild with acceptable safety profile.

[Evaluation of training course of hepatic disease in grass roots clinical hepatic physicians in some poverty-stricken counties in Shanxi and Shaanxi provinces].

Objective: To understand the awareness of hepatic disease related knowledge among hepatic physicians in poverty-stricken counties in China, assess the effectiveness of training and provide a reference for the training in the future. Methods: The training was conducted in 90 clinical hepatic physicians selected from county hospitals in poverty-stricken counties (or cities) in Shanxi and Shaanxi provinces. An examination was conducted before the training, immediately after the training and at 5(th) month after the training, respectively. One-way analysis of variance and χ(2) test were conducted to evaluate the score and the correct rate. Results: The knowledge score was (42.96±14.02) before the training, (62.86±13.28) immediately after the training and (59.03±17.92) at 5(t)h month after the training, and the differences were significant. After the training, the awareness of all aspects of related knowledge was improved, the difference was significant compared to knowledge score before training, and at 5(th) month after the training, the difference was still significant. Conclusion: After the training, the awareness of liver disease related knowledge of clinical hepatic physicians in poverty-stricken counties (cities) in Shanxi and Shaanxi provinces was improved, and the improvement could be maintained for nearly half a year.

Screening of differentially expressed genes in pathological scar tissues using expression microarray.

Pathological scar tissues and normal skin tissues were differentiated by screening for differentially expressed genes in pathologic scar tissues via gene expression microarray. The differentially expressed gene data was analyzed by gene ontology and pathway analyses. There were 5001 up- or down-regulated genes in 2-fold differentially expressed genes, 956 up- or down-regulated genes in 5-fold differentially expressed genes, and 114 up- or down-regulated genes in 20-fold differentially expressed genes. Therefore, significant differences were observed in the gene expression in pathological scar tissues and normal foreskin tissues. The development of pathological scar tissues has been correlated to changes in multiple genes and pathways, which are believed to form a dynamic network connection.

Total blood volume of Asian patients undergoing cardiac surgery is far from that predicted by conventional methods.

AIM: Current cardiopulmonary bypass (CPB) procedures use non-hematic fluids to prime bypass circuits, often resulting in marked hemodilution. Patients' total blood volume (TBV) is estimated prior to hemodilution. We aimed to evaluate differences between calculation of TBV by Nadler's formula, a classic reference book method, and an established formula calculated by the authors. METHODS: A total of 285 patients of Asian origin received primary cardiac surgery between September 2010 and October 2011 in our institution. Patients' total blood volume was estimated by: 1) standard Nadler formula: TBV (men) =0.417H3+0.045TBM-0.030L TBM (total body mass, Kg); TBV (women) =0.414H3+0.0328 TBM-0.030L; 2) classic reference book method: patient's weight in kilograms times 7% (women) or 7.5% (men); and 3) our practical calculation: TBV=HCT2*(CPB prime volume + intravenous fluids before CPB - urine volume before CPB)/(HCT1- HCT2). RESULTS: Bland-Altman plotting revealed no mean differences between Nadler formula and reference book TBV measurements (Figure 1A). Differences in means (95% limit of agreement) for reference book/Nadler formulas was 0.52 (-0.21, 1.24, N.=285). Comparing authors' results with those of reference book/Nadler, TBV yielded divergent results. TBV correlated positively to patient's height (P=0.001) and body surface area (P<0.01), and correlated positively to height after controlling for age and gender (ß=87.3, SE=42.9, P=0.043). CONCLUSION: Total blood volume of Asian patients calculated by the authors differs markedly from that estimated by Nadler and classic reference book formulas, which suggests that more accurate calculation of TBV is needed for Asian cardiac patients requiring CPB, especially patients with valvular disease.

Scavenger receptor class B type I mediates cell entry of hepatitis C virus.

This study assessed the functional role of human scavenger receptor class B type I (SR-BI) as a putative hepatitis C virus (HCV) receptor using Chinese hamster ovary (CHO) cells transfected with human SR-BI (CHO-huSR-BI). The expression of SR-BI by primary Tupaia hepatocytes (PTHs), human hepatocarcinoma cell line (HepG2) cells, untransfected CHO cells and CHO-huSR-BI cells was analysed by Western blotting. Receptor competition assays showed that anti-SR-BI antibodies that block the binding of soluble envelope glycoprotein E2 could prevent HCV infection. Pre-incubation of CHO-huSR-BI and HepG2 cells with anti-SR-BI antibodies resulted in marked inhibition of E2 binding. After incubation with HCV RNA-positive serum from a patient with chronic HCV infection, however, HCV infection could not be detected in CHO-huSR-BI cells, but was detected in PTHs. These results demonstrate that, whilst SR-BI represents an important cell surface molecule for HCV infection, the presence of SR-BI alone is insufficient for HCV entry.

In vitro hepatic differentiation of mesenchymal stem cells from human fetal bone marrow.

We examined whether human fetal mesenchymal stem cells (FMSCs) derived from fetal bone marrow were able to differentiate into functional hepatocyte-like cells in vitro. The surface phenotype of FMSCs was characterized by flow cytometry. To induce hepatic differentiation of FMSCs, we added hepatocyte growth factor, basic fibroblast growth factor and oncostatin M into the cell culture medium. After 21 days of hepatocyte induction, FMSCs expressed the hepatocyte-specific markers, alpha-fetoprotein and cytokeratin 18, as demonstrated by immunofluorescence staining. Differentiated FMSCs also demonstrated in vitro functions characteristic of liver cells, including albumin production, urea secretion and glycogen storage. In conclusion, fetal bone marrow-derived FMSCs are able to differentiate into functional hepatocytelike cells and may serve as a source of cells for liver disease therapy.

Is benzene oxide homoaromatic? A microcalorimetric study.

Rate constants and heats of reaction for the aromatization of benzene oxide (1) and the acid-catalyzed aromatization of benzene hydrate (2) in highly aqueous solution giving phenol and benzene, respectively, have been measured by heat-flow microcalorimetry. The measured heat of reaction of benzene oxide, DeltaH = -57.0 kcal mol(-1), is much larger than that of benzene hydrate, DeltaH = -38.7 kcal mol(-1), despite an unusually low reactivity of benzene oxide, rate ratio 0.08. The measured enthalpies agree with those calculated using the B3LYP hybrid functional corrected with solvation energies derived from semiempirical AM1/SM2 calculations. Comparison with the measured enthalpies of the corresponding reactions of the structurally related 1,3-cyclohexadiene oxide (3) and 2-cyclohexenol (4) of DeltaH = -24.9 kcal mol(-1) (includes a small calculated correction of -1.2 kcal mol(-1)) and DeltaH approximately 0 kcal mol(-1), respectively, gives a smaller aromatization energy for the benzene oxide than for the benzene hydrate reaction (DeltaDeltaDeltaH = 6.6 kcal mol(-1)). This suggests that benzene oxide is unusually stabilized by a significant amount of homoaromatization as has been proposed previously (J. Am. Chem. Soc. 1993, 115, 5458). This unusual stability accounts for more than half of the approximately 10(7) times lower than expected reactivity of benzene oxide toward acid-catalyzed isomerization. The rest is suggested to originate from an unusually high energy of the carbocation-forming transition state.

[Prolongation of survival of skin allograft with anti-Thy-1 monoclonal antibody in mice].

Skin allograft rejection is mainly mediated by T lymphocytes. We investigated the cytotoxicity of anti-Thy-1 McAb to murine thymocytes in vitro, and evaluated the prolonging effect on the survival of murine skin allografts (BALB/c--C57BL/6) in vivo with the McAb. The results showed murine thymocytes were destroyed in vitro by the McAb with complement; the survival of skin allograft was prolonged in vivo with the McAb. Lymphocyte infiltration in skin allografts was inhibited. These results provide a valuable reference for the clinical usefulness of anti-CD3 McAb in prolonging survival of human skin allografts in burn patients.