Objective To investigate whether γH2AX could be a useful biomarker for evaluating the DNA double‐stranded . Methods Semem samples in case group were from 27 infertile males who were diagnosed in Andriatrics department or reproductive centre in the First Affiliated Hospital of Guangxi Medical University .The other semen samples were from 23 healthy donors with fertility as comparison .The levels of γH2AX were detected by flow cytometry .Single cell gel electropherosis(SCGE)was applied to assess the level of DSBs of sperm .Density gradient centrifugation(DGC) was applied to optimized spermatozoa .Results TheγH2AX levels and the DSBs of the sperm of the infertile subjects were significantly higher than those of healthy males(P<0 .01) , and the levels of γH2AX and the DSBs of sperm significantly decreased in two groups by DGC(P<0 .01) .Conclusion The level of spermatozoaγH2AX is higher in male infertility patients than in healthy donors with fertility ,which might be a useful biomarker for evaluating DSBs of sperm .

Objective To evaluate the effects of propofol on the proliferation of hippocampal neurons in fetal rats in vitro.Methods Pregnant Sprague-Dawley rats at 16-18 days of gestation,were sacrificed and the fetal rats were taken out from the abdominal cavity.The hippocampal neurons of the fetal rats were isolated and seeded in culture plates.After being cultured for 9 days,the neurons were divided into 7 groups using a random number table(n =36 each):control group (C group),intralipid group (I group) and propofol 0.1,1.0,10.0,100.0,1 000.0 μmol/L groups (P1-5 groups).In group I,10％ intralipid was added to the culture media until the final concentration reached 100 μmol/L.In P1-5 groups,propofol was added to the culture media until the final concentration reached 0.1,1.0,10.0,100.0 and 1 000.0 μmol/L,respectively.The neurons were then incubated for 3 h.The proliferation of hippocampal neurons was determined by MTT assay at 12,24,36,48,60 and 72 h after the end of incubation with propofol.Results Compared with group C,the proliferation of hippocampal neurons was significantly decreased in P1-5 groups (P ＜ 0.05),while no significant change was found in group I (P ＞ 0.05).Compared with group P1,the proliferation of hippocampal neurons was significantly decreased in group P5 (P ＜ 0.05),while no significant change was found in P2-4 groups (P ＞ 0.05).Conclusion Propofol can decrease the proliferation of hippocampal neurons in fetal rats in vitro.

Objective To explore the effect of interferon-?(IFN-?)on phenotypic transition of human Tenon’s conjunctival capsular fibroblast(HTCF). Methods Cultured HTCF derived from 4 operated human cataracts was induced for 48 hours in absence or presence of IFN-? and/or transforming growth factor-?1(TGF-?1). Then immunocytochemistry and Western blot technology were used to detect the ?-smooth muscle actin(?-SMA)expression and identificate the cell phenotype. Results In contrast to normal HTCF, IFN-?(10 ng/mL) inhibited the expression of ?-SMA(P

Objective To evaluate the effect of dexmedetomidine on the activity of glycogen synthase kinase-3 beta (GSK-3β) during propofol-induced apoptosis in hippocampal nerve cells of newborn rats.Methods Sixty male 7-day-old Sprague-Dawley rats,weighing 10-15 g,were divided into 6 groups (n=10 each) using a random number table:normal saline group (group NS),fat emulsion group (group F),propofol group (group P) and different doses of dexmedetomindine groups (group D25,group D50 and group D75).Normal saline and fat emulsion 100 μl were injected intraperitoneally in group NS and group F,respectively.In group P,propofol 50 mg/kg was intraperitoneally injected,and an increment of propofol 50 mg/kg was added after righting reflex completely recovered,with the total amount of 100 mg/kg.In group D25,group D50 and group D75,dexmedetomidine 25,50 and 75 μg/kg were intraperitoneally injected,respectively,and 30 min later propofol 100 mg/kg was administered.At 2 h after emergence,the rats were sacrificed,their brains were removed for determination of apoptosis in hippocampal nerve cells (by TUNEL),and the hippocampi were isolated for detection of the expression of GSK-3β and phosphorylated GSK-3β (p-GSK-3β) by Western blot analysis.The apoptosis index (AI) and ratio of p-GSK-3β/GSK-3β were calculated.Results Compared with group NS,AI was significantly increased,the expression of p-GSK-3β was down-regulated,and the p-GSK-3β/GSK-3β ratio was decreased in P,D25,D50 and D75 groups (P＜0.05).Compared with group P,AI was significantly decreased,the expression of p-GSK-3β was up-regulated,and the p-GSK-3β/GSK-3β ratio was increased in D25,D50 and D75 groups (P＜0.05).Compared with group D25,AI was significantly decreased (P＜0.05),and no significant change was found in the expression of p-GSK-3β or ratio of p-GSK-3β/GSK-3β in D50 and D75 groups (P＞0.05).Compared with group D50,AI was significantly decreased (P＜0.05),and no significant change was found in the expression of p-GSK-3β or ratio of p-GSK-3β/GSK-3β in group D75 (P＞0.05).Conclusion The mechanism by which dexmedetomidine attenuates propofol-induced apoptosis in hippocampal nerve cells may be related to inhibition of GSK-3β activity in newborn rats.

Objective To evaluate the long-term efficacy of recombinant human endostatin (rh-Endo) combined with FOLFOX4 as an adjuvant treatment for patients of stage Ⅱ and Ⅲ colorectal cancer.Methods Eligible patients were randomly assigned to receive FOLFOX4 or FOLFOX4 plus rh-Endo regimen in which patients receiving 7.5 mg/m2 Ⅳ on day 1-7,repeated every 2 weeks,to a total of 12 cycles in 6 months.Results A total of 197 eligible patients were accrued in this research with 105 patients in the control group and 92 patients in the experimental arm.Median follow-up period was 42 months.The baseline characteristics distributed were balanced by treatment.Rh-Endo combined with FOLFOX4 regimen resulted in significant improvement on DFS compared to FOLFOX4 regimen for patients with stage Ⅲ colon cancer (HR =0.19,95％ CI0.05-0.75,P =0.0124),and with a 34％ improvement on 3-year DFS and 81％ reduced recurrence.Although rh-Endo combined with FOLFOX4 regimen failed to make significant difference on DFS in the whole (HR =0.75,95％ CI 0.31-1.83,P =0.5589),it was also observed a 17％ improveiment on 3-year DFS.No statistical significant difference on DFS was observed in patients with stage Ⅱ disease.Conclusions Rh-Endo combined with FOLFOX4 regimen significantly improved the disease-free survival for patients with stage Ⅲ colorectal cancer,indicating that patients with stage Ⅲ disease,but not stage Ⅱ disease,can benefit from FOLFOX4 plus rh-Endo regimen in adjuvant treatment.

Objective To establish and evaluate a gene microarray for determination katG mutations of Mycobacterium tuberculosis isolates associated with resistance to isoniazid(INH).Methods A panel of probes were designed and gene chips were prepared by dotting.Mycobacterium tuberculosis isolates resistance to 5 drugs was determined by proportional dilution methods.Amplicons of Mycobacterium tuberculosis isolates were detected by our chip and sequenced.Results The drug resistance rate of the isolates to at least one of the anti-tuberculosis drugs was 70.8%(97/137).45 strains out 137 Mycobacterium tuberculosis isolates was resistant to INH(32.8%).katG was successfully amplified from 100% of the susceptible strains and 88.9%(40/45)resistant strains.4 of 45 INH resistant isolates' katG were deleted.27 of 40(67.5%) katG has been detected to have katG 315 codon mutations.The mutations were 315 AAC(Asn,13/40), ACC(Thr,6/40),ACA(Thr,4/40),ATC(Ile,2/40),AGC(Arg,2/40).The mutation rate of katG analyzed by gene chips we prepared were identical to katG sequencing.Conclusion The gene microarray techniques we developed for determination of Mycobacterium tuberculosis resistance to INH are specific, sensitive and may be used as an alternative in clinical laboratory.

The compatibility of traditional Chinese medicines (TCMs) formulae containing enormous information, is a complex component system. Applications of mathematical statistics methods on the compatibility researches of traditional Chinese medicines formulae have great significance for promoting the modernization of traditional Chinese medicines and improving clinical efficacies and optimizations of formulae. As a tool for quantitative analysis, data inference and exploring inherent rules of substances, the mathematical statistics method can be used to reveal the working mechanisms of the compatibility of traditional Chinese medicines formulae in qualitatively and quantitatively. By reviewing studies based on the applications of mathematical statistics methods, this paper were summarized from perspective of dosages optimization, efficacies and changes of chemical components as well as the rules of incompatibility and contraindication of formulae, will provide the references for further studying and revealing the working mechanisms and the connotations of traditional Chinese medicines.
Chemistry, Pharmaceutical ; statistics & numerical data ; Data Interpretation, Statistical ; Drug Incompatibility ; Drugs, Chinese Herbal ; analysis ; Medicine, Chinese Traditional

Objective To study the characteristics of clustering of cardiovascular risk factors in patients less than 50 years-old of premature stable coronary heart disease(PSCHD)complicated with nonalcoholic fatty liver(NAFL).Methods One hundred and six patients with documented PSCHD were recruited into this study and their clinical data,including biochemical parameters,high-sensitivity C-reactive protein(hsCRP),white blood cell(WBC)count,ete.,were analyzed based on whether they had NAFL by B-type ultrasound scanning and their homeostasis model assessment ratio(Homa-IR)by the criteria for metabolic syndrome formulated by the International Diabetes Federation.Results Thirty-two (30.1percent)of 106 patients of PSCHD complicated with NAFL,and 74(69.9 percent)without NAFL. As compared to patients without NAFL,patients with NAFL had higher fasting blood glucose(FBS),serum level of insulin(INS),total cholesterol(TC),triglyceride(TG),serum activity of alanine aminotransferase(ALT),hsCRP,WBC count,body mass index(BMI),Homa-IR,and higher proportion of those with abnormal blood glucose,hypertension.metabolic syndrome(MS)and carotid atherosclerosis (CA)(P＜0.05),respectively.Bi-variate correlation analysis revealed that hsCRP positively correlated to BMI,TG,ALT and Homa IR(r=0.420,P=0.000;r=0.200,P=0.040;r=0.218,P=0.048:and r=0.546,P=0.000,respectively)and inversely correlated with serum level of high-density lipoprotein cholesterol(HDL-C)(r=-0.220,P=0.023).WBC count positively correlated with FBS(r=0.211,P=0.030).BMI,hsCRP,ALT,and proportions of hypertension,diabetes,MS,NAFL and CA in patients with Homa-IR above median were significantly higher than those in patients with that below median ( P＜0.05,respectively).Conclusions More risk faetors for chronic inflammatory reaction,cardiovascular disease and insulin resistance were clustered more obviously in patients of PSCHD complicated with NAFL.

Objective To investigate the mechanism of BVT. 2733 on insulin resistance, by using diet-induced obese (DIO) mice model. Methods After having been balanced for 3 days, the C57BL/6J mice were randomly divided into a normal diet group and a high-fat diet (HFD) group. After 20 weeks, the obese mice were further randomly divided into an obese control group, a BVT. 2733 group and a pioglltazone (PGZ) group and they were orally administered with placebo, BVT. 2733 and PGZ separately for two weeks.Adiponectin and leptin mRNA expression levels from adipose tissue were analyzed with real-time quantitative PCR. The levels of plasma glucose, serum insulin and adiponectin were measured with biochemical technology, radioimmunoassay and ELISA. Adipocyte sizes were observed with immunohistocbemistry.Results The body weight, plasma glucose and serum insulin levels raised(P<0.05)in the HFD group and the adipocyte sizes were bigger. Serum insulin levels significantly reduced (P<0.05) and adipocyte sizes reduced, while plasma adiponectin level raised (P<0.01)in the two treatment groups as compared with those in obese controls. Both the mRNA expressions of adiponectin and leptin upregulated(P<0.05)in the PGZ group, but their expressions in the BVT. 2733 group did not alter significantly. The body weight of the mice reduced significantly in the BVT. 2733 group. Conclusion BVT. 2733 can reduce body weight significantly and improve insulin resistance, but cannot influence the expression of adipocytokines.

Objective To evaluate the role of opioid receptors in fentanyl-induced inhibition of proliferation and migration of human gastric cancer cell line MGC-803.Methods The human gastric cancer cell line MGC-803 was cultured in DMEM liquid culture medium.The cells were seeded in 6-well or 96-well plates and then randomly divided into 4 groups (n =54 each):control group (group C),fentanyl group (group F),naloxon group (group N) and naloxon + fentanyl group (group NF).The cells were exposed to 0.1 μmol/L fentanyl and 10 μmol/L naloxon in F and N groups,respectively.The cells were incubated with 10 μmnol/L naloxon for 30 min and then O.1 μmol/L fentanyl was added to the culture medium in group NF.The viability of the cells was detected by MTT assay after being incubated with fentanyl for 12,24,36,48,60 and 72 h.The cell apoptosis was assessed by flow cytometry after being incubated with fentanyl for 24 h.The migration of the cells was detected by wound healing assay after being incubated with fentanyl for 48 h.The proliferation of the cells was determined by colony formation assay at 7 day of incubation with fentanyl.Results Compared with group C,no significant changes in the viability of the cells,rate of colony formation,apoptotic rate and rate of cell wound healing were found in group N (P ＞ 0.05),and the viability of the cells,rate of colony formation and rate of cell wound healing were significantly decreased,and the apoptotic rate was increased in F and NF groups (P ＜ 0.05).There was no significant difference in the viability of the cells,rate of colony formation,rate of cell wound healing and apoptotic rate between group NF and group F (P ＞ 0.05).Conclusion Opioid receptors are not involved in fentanyl-induced inhibition of proliferation and migration of human gastric cancer cell line MGC-803 in vitro.