Cytotoxic effects of Coptis chinensis and Epimedium sagittatum extracts and their major constituents (berberine, coptisine and icariin) on hepatoma and leukaemia cell growth

1. The present study was conducted to evaluate the cytotoxic effects of Coptis chinensis and Epimedium sagittatum extracts and their major constituents on hepatoma and leukaemia cells in vitro. 2. Four human liver cancer cell lines, namely HepG2, Hep3B, SK-Hep1 and PLC/PRF/5, and four leukaemia cell lines, namely K562, U937, P3H1 and Raji, were used in the present study. 3. Of the two crude drugs, C. chinensis exhibited the strongest activity against SK-Hep1 (IC50 = 7 microg/mL) and Raji (IC50 = 4 microg/mL) cell lines. The IC50 values for C. chinensis on HepG2, Hep3B and PLC/PRF/5 cell lines were 20, 55 and 35 microg/mL, respectively. The IC50 values for C. chinensis on K562, U937 and P3H1 cell lines were 29, 29 and 31 microg/mL, respectively. 4. With the exception of HepG2 and Hep3B, the E. sagittatum extract inhibited the proliferation of all cell lines (SK-Hep1, PLC/PRF/5, K562, U937, P3H1 and Raji), with IC50 values of 15, 57, 74, 221, 40 and 80 microg/mL, respectively. 5. Interestingly, the two major compounds of C. chinensis, berberine and coptisine, showed a strong inhibition on the proliferation of both hepatoma and leukaemia cell lines, with IC50 values varying from 1.4 to 15.2 microg/mL and from 0.6 to 14.1 microg/mL, respectively. However, icariin (the major compound of E. sagittatum) showed no inhibition of either the hepatoma or leukaemia cell lines. 6. The results of the present study suggest that the C. chinensis extract and its major constituents berberine and coptisine possess active antihepatoma and antileukaemia activities.     

钉螺线粒体基因组DNA提取方法的优化及比较

目的优化提取钉螺线粒体基因组DNA(mt DNA)的方法。方法应用高温裂解、蛋白酶K变温消化和醋酸钾纯化对传统的高盐沉淀法进行优化,并将优化的高盐沉淀法与蔗糖密度梯度离心法、传统的高盐沉淀法进行比较。用紫外分光光度计、琼脂糖凝胶电泳和线粒体COX1基因PCR扩增产物鉴定提取的mt DNA,并用核ITS基因PCR扩增产物检测有无核DNA污染。结果优化的高盐沉淀法获得的mt DNA浓度和产量较高,差异有统计学意义(F=3 032.65、10185.00,P均0.01)。用凝胶电泳检测后未发现核DNA与蛋白质污染,质量可满足于PCR、测序等分子生物学研究。结论优化的高盐沉淀法简易高效,成本低,获得的mt DNA能满足相关分析要求。     

New progress in understanding roles of nitric oxide during hepatic ischemia-reperfusion injury

Hepatic ischemia-reperfusion injury (HIRI) is a major clinical cause of morbidity and mortality in liver surgery and transplantation. Many studies have found that nitric oxide (NO) plays an important role in the HIRI and its increase or decrease can affect the progression and outcome of HIRI. However, the role of NO in HIRI is controversial and complicated. NO derived by endothelial NO synthase (eNOS) shows a protective role in HIRI, while excessive NO derived by inducible NO synthase (iNOS) accelerates inflammation and increases oxidative stress, further aggravating HIRI. Nevertheless, the overexpression of eNOS may exacerbate HIRI and iNOS-derived NO in some cases reduces HIRI. Here we review the new progress in the understanding of the roles of NO during HIRI: (1) NO possesses different roles in HIRI by increasing NO bioavailability, down-regulating leukotriene C4 synthase, inhibiting the activation of the nuclear factorκB (NFκB) pathway, enhancing cell autophagy, and reducing inflammatory cytokines and reactive oxygen species (ROS). And NO has both protective and deleterious effects by regulating apoptotic factors; (2) eNOS promotes NO production and suppresses its own overexpression, exerting a hepatoprotective effect reversely. Its activation is regulated by the PI3K/Akt and KLF2/AMPK pathways; and (3) iNOS derived NO mainly has deteriorating effects on HIRI, while it may have a protective function under some conditions. Their expression should reach a balance to reduce the adverse side and make NO protective in the treatment of HIRI. Thus, it can be inferred that NO modulating drugs may be a new direction in the treatment of HIRI or may be used as an adjunct to mitigate HIRI for the purpose of protecting the liver.     

Chemoprevention against hepatocellular carcinoma of Cornus officinalis in vitro

The water extracts of Cornus officinalis Sieb. et Zuce against hepatocellular carcinoma (HCC) was studied for its chemopreventive potential. Three HCC cell lines (HepG2, SK-Hep1 and PLC/PRF/5) and three leukemic cell lines (U937, K562 and Raji) were tested with XTT assay. Extracts of C. officinalis inhibited all these HCC cells and leukemic cells at a concentration of 100 microg/ml (P < 0.05) and was dose-dependent (P < 0.0001). P53 (P< 0.0001) and Ras (P = 0.001) significantly affected its activity against HCC. Extracts of C. officinalis also possessed the anti-oxidant activity through free radicals scavenging activity at a concentration of 50 microg/ml (P < 0.05). In summary, our experiment implied that C. officinalis might be a candidate for chemopreventive agent against HCC through the antioxidant and anti-neoplastic effects.     

Percutaneous transgastric endoscopic tube ileostomy in a porcine survival model

AIM To introduce natural orifice transgastric endoscopic surgery(NOTES) tube ileostomy using pelvis-directed submucosal tunneling endoscopic gastrostomy and endoscopic tube ileostomy.METHODS Six live pigs(three each in the non-survival and survival groups) were used. A double-channeled therapeutic endoscope was introduced perorally into the stomach. A gastrostomy was made using a 2-cmtransversal mucosal incision following the creation of a 5-cm longitudinal pelvis-directed submucosal tunnel. The pneumoperitoneum was established via the endoscope. In the initial three operations of the series, a laparoscope was transumbilically inserted for guiding the tunnel direction, intraperitoneal spatial orientation and distal ileum identification. Endoscopic tube ileostomy was conducted by adopting an introducer method and using a Percutaneous Endoscopic Gastrostomy Catheter Kit equipped with the Loop Fixture. The distal tip of the 15 Fr catheter was placed toward the proximal limb of the ileum to optimize intestinal content drainage. Finally, the tunnel entrance of the gastrostomy was closed using nylon endoloops with the aid of a twin grasper. The gross and histopathological integrity of gastrostomy closure and the abdominal wall-ileum stoma tract formation were assessed 1 wk after the operation.RESULTS Transgastric endoscopic tube ileostomy was successful in all six pigs, without major bleeding. The mean operating time was 71 min(range: 60-110 min). There were no intraoperative complications or hemodynamic instability. The post-mortem, which was conducted 1-wk postoperatively, showed complete healing of the gastrostomy and adequate stoma tract formation of ileostomy.CONCLUSION Transgastric endoscopic tube ileostomy is technically feasible and reproducible in an animal model, and this technique is worthy of further improvement.     

Effects of apoptosis on liver aging

As an irreversible and perennial process,aging is accompanied by functional and morphological declines in organs. Generally,aging liver exhibits a decline in volume and hepatic blood flow. Even with a preeminent regenerative capacity to restore its functions after liver cell loss,its biosynthesis and metabolism abilities decline,and these are difficult to restore to previous standards. Apoptosis is a programmed death process via intrinsic and extrinsic pathways,in which Bcl-2 family proteins and apoptosis-related genes,such as p21 and p53,are involved.Apoptosis inflicts both favorable and adverse influences on liver aging.Apoptosis eliminates transformed abnormal cells but promotes age-related liver diseases,such as nonalcoholic fatty liver disease,liver fibrosis,cirrhosis,and liver cancer. We summarize the roles of apoptosis in liver aging and age-related liver diseases.     

Stereoselective Disposition of Suspensions of Conventional and Wax-matrix Sustained Release Ibuprofen Microspheres in Rats

Purpose. The aim of the study is to evaluate stereoselective in vivo disposition of suspensions of conventional and wax-matrix sustained release ibuprofen microspheres in rats. Methods. Male wistar rats were dosed IV with 20 mg/kg, or orally with conventional suspension, and three suspension formulations of sustained release microspheres (having three different particle sizes) of racemic ibuprofen. Blood samples were analyzed stereoselectively by reverse phase HPLC. Results. The mean C_max for (S)- and (R)- ibuprofen decreased with increased particle size of the drug or microspheres in the suspension dosage forms, while the T_max increased with increased particle size. The mean S/R ratio (AUC_0–48) of the suspensions decreased with increase in particle size of the drug or microspheres and these ratios (for both conventional and sustained formulations) were higher than that of the IV, an indication of presystemic inversion. Decrease in the ratios with increased particle size is suggestive of formulation dependent inversion. The plasma concentration-time data of the sustained release formulations showed bimodal profiles, irrespective of the particle size of the microspheres. The second peak observed after 8 hours is indicative of colonic absorption. Conclusions. Stereoselective disposition of ibuprofen microspheres showed higher bioavailability compared to the conventional suspension. Bimodal disposition is influenced by dosage form while presystemic inversion is both site-specific, and dosage form dependent.     

The associations between menstrual function and life style/working conditions among nurses in Taiwan

This study was aimed at understanding the relationship among menstrual pattern, dysmenorrhea, life style and working conditions in nurses. The nurses were randomly selected from a medical center in Northern Taiwan. Each subject completed daily records including life and working conditions during the study period. The study showed that there were statistically significant differences in work years, daily working hours and type of work shift among nurses that worked at different units in the hospital. In the perceived regular cycle group, nurses that worked the night shift only exhibited the shortest menstrual cycles, less than 25 d. There was a significant difference (p<0.05) among the nurses' menstrual cycles. Many life factors (such as passive tobacco smoke exposure, perceived life satisfaction and perceived life stress) and working factors (such as work years, perceived work satisfaction and perceived work stress) were not significantly related to menstrual cycle regularity. In addition, 30% of the nurses complained of dysmenorrhea. Some factors including age, marital status and perceived life satisfaction were significantly related to dysmenorrhea. However, other life factors (such as passive tobacco smoke exposure, smoking, coffee, alcohol, cold drink habits, exercise and perceived life stress) and working factors (such as working places, type of work shift, daily work hours, perceived work satisfaction and perceived work stress) showed no correlation with dysmenorrhea. This study indicates that women should pay attention to their menstrual function and dysmenorrhea phenomenon.     

Effectiveness of telephone follow-up and counseling in aftercare for alcoholism

This study investigates the effectiveness of telephone follow-up and counseling for alcoholism outpatients on abstinence, readmission, alcohol consumption, addiction severity and adjustment at three months follow-up. A quasi-experimental pre-post control group design was used. Subjects were recruited from a psychiatric center from March to September 2001 by purposive sampling. 34 cases were studied in each of an experimental and a control group. The structured instruments included an Addiction Severity Index (ASI) and a Progress Evaluation Scale (PES). Data were analyzed using t-test, chi-square, and generalized estimating equation regression analyses. Abstinence rate (p <.05) and adjustment (p <.05) at three months post-discharge were significantly increased in the experimental group. Addiction severity (p <.001) and readmission rate (p <.05) were significantly decreased, whereas decrease in alcohol consumption was not significant. These results showed that the telephone follow-up intervention contributed to better outcomes and demonstrated the importance of aftercare for alcoholism. It is suggested that developing a rehabilitation treatment program should enhance abstinence for alcoholism.