Objective To investigate the applieation of restriction display (RD) technique in the preparation of HCV probes of clinical genotyping microarray. Methods Restriction enzyme Sau3A Ⅰwas chosen to digest the full-length HCV cDNAs of three distinct subtypes, i.e.1a, 1b and 2a. The resultant restrictive fragments were then ligated with universal adapters. PCR primers were designed to match the universal adapters but with one "nesting" base overhanging at the 3′- end. The PCR reactions were performed by ten pairs of different primer combinations. The differential genes were separated through polyacrylamide gel electrophoresis and silver staining. The second-round PCR was performed using the isolated bands as PCR templates. The purified PCR products were then cloned into T-vectors. The recombinant plasmids were extracted from positive recombinant clones and the target gene fragments were sequenced. Results The target HCV gene fragments ranging from 200 to 900bp were isolated and sequenced, which were correlated precisely with the RFLP (Restriction Fragment Length Polymorphism) prediction. A total of 66 different fragments were obtained, averaging about 22 for each subtypes. These fragments could be further used as probes in HCV microarray preparations. Conclusion RD technique is of great value in obtaining a large number of equal sized gene probes, which provide a swift protocol in generating DNA probes for the preparation of microarrays.

According to the teaching practice in biochemistry laboratory course,we describe the characteristics of international students'teaching,teaching preparation,teaching course and so on.These experiences may provide an important source of information for teaching practice in the future.

Objective To investigate the survival rates of main malignant tumors in Nanhui district. Methods Total 5 783 cases, registered as new cases with malignant neoplasm from 2002 to 2004 in Nanhui, were followed up. Data was studied by survival analysis with SPSS 13.0 of life table and Kaplan -Meier. Results The average annual crude incidence of malignant tumors was 267.57 / 100 000, the World standardized incidence rate of 209.98 / 100 000. Cancer Incidence in the top five ranks followed by lung, liver, stomach, colon and esophageal cancer, the incidence was 43.06 / 100 000, 28.65 / 100 000, 26.82 / 100 000, 18.63 / 100 000 and 9.57 / 100 000, respectively. The difference of the survival rates between the top five cancers was remarkable. Survival rates of cancers among males from the lowest to the highest were liver, lung, esophagus, stomach and intestines. Survival rates of cancers among females from the lowest to the highest were of liver, lung, esophagus, stomach, intestines, cervical and breast. The male or female main cancer's relative survival rate droped along with the age rising. Conclusion The survival status was better in the patients with intestine cancers and female patients with breast cancer, but worse in patients with liver, ung and esophageal cancer.

Objective: To investigate the awareness of acquired immune deficiency syndrome (AIDS) knowledge and AIDS-related behaviors among youth students in gay dating sites, and to provide evidences for AIDS prevention education through the internet.Methods: The students in gay dating sites, selected by a snowball sampling, were interviewed by questionnaires.Chi-square tests were used to analyze the awareness of AIDS knowledge among the students of different characteristics.The Logistic regression was used to analyze the factors associated with ever testing for HIV.Results: In the study, 469 youth students in gay dating sites filled in the questionnaires, and a total of 442 (94.2%) valid samples were collected.The awareness of HIV/AIDS knowledge for the public among the youth students in gay dating sites was 83.9% (371).The awareness of HIV/AIDS knowledge for the youth students was 77.1% (341), and the rate of ever testing for HIV was 52.0% (230).The awareness of HIV/AIDS knowledge for the public in the students were different in different age groups (P=0.001), different marital statuses (P<0.001), different sexual orientations (P<0.001), and different genders of the first sexual partner (P<0.001).The awareness of HIV/AIDS knowledge for the youth students were different in different age groups (P=0.010), different marital status (P=0.004), different sexual orientations (P<0.001), and different genders of the first sexual partner (P<0.001).The rate of ever having sexual intercourse among the youth students in gay dating sites was 75.1% (332), and the rate of multiple sexual partnerships among the youth students was 41.3% (137).Compared with homosexual orientation, sexual orientation as heterosexual (OR=0.282,95%CI:0.151 to 0.528) and not sure (OR=0.175,95%CI: 0.035 to 0.885) were risk factors of ever testing for HIV.Multiple sexual partnerships (OR=2.103,95%CI:1.278 to 3.462) were promoting factors of ever testing for HIV.Conclusion: The rate of high-risk behaviors among the youth students in gay dating sites was high.The concern should be raised to heterosexual male students who had tendency to homosexual behavior.The AIDS prevention education should be developed in gay dating sites, to improve the self-protection awareness of the youth students.

Objective To analyze the prevention and treatment of postoperative hemorrhage after pancreaticoduodenectomy. Methods The clinical data of 142 patients undergoing pancreaticoduedenectomy from Jan 1995 to Dec 2008 were retrospectively analyzed. Results The incidence of postoperative hemorrhage was 14.1% (20/142), the mortality caused by this complication was 35% (7/20). Among these patients, intra-abdominal hemorrhage occurred in seven cases, and gastrointestinal hemorrhage occurred in 14 cases, with one case suffering both. There were three and four cases of early and delayed intra-abdominal hemorrhage respectively. Early and delayed gastrointestinal hemorrhage occurred in three and eleven cases respectively. Univariate analysis showed that operative blood loss, blood transfusion, infection and pancreatic fistula were significantly associated with postoperative hemorrhage. Multivariate analysis using Logistic regression identified two variables as independent factors associated with postoperative hemorrhage, namely, infection(OR=6.918) and pancreatic fistula(OR=3.948). Conclusions The incidence and mortality of hemorrhage after pancreaticoduodenectomy is still high. Skillful operation and prevention of pancreatic fistula, infection and stress ulcer are the key paints for reducing postoperative hemorrhage. Proper treatments should be used according to the site, onset and severity of hemorrhage.

ObjectiveTo study the teaching method of childbirth turning point and enhances teaching effect in the normal birth of the gynecology and obstetrics department teaching. MethodsAfter carting on the different form the teaching, to 382 students of Qujing medical high school, we carry on the one-to-one skill operation inspection and make the contrastive analysis of the achievement test scores of the students in the experimental group and the control group. Results There is remarkable difference between the conventional experiment teaching and the experiment teaching of adding giant pelvis ( P＜0.01 ). ConclusionThe giant pelvis's application can enhance the teaching effect enormously in the childbirth turning point teaching, and this method has the application and the promoted value.

AIM:To investigate the role of peroxisome proliferator-activated receptor β( PPARβ)-nitric oxide (NO) signal pathway in cardiomyocyte hypertrophy induced by high glucose (25.5 mmol/L) and insulin (0.1 μmol/L) ( HGI) .METHODS: The cardiomyocyte hypertrophy was characterized in rat primary cardiomyocytes by measuring the cell surface area, protein content, and the mRNA expression of atrial natriuretic factor (ANF).The mRNA and protein ex-pression were measured by real-time PCR and Western blotting , respectively .The activity of NO synthase ( NOS) and NO content were measured by a reagent kit through ultraviolet spectroscopy .RESULTS:HGI induced profound change of hy-pertrophic morphology , and significantly increased the cell surface area , protein content and mRNA expression of ANF (P<0.01), but decreased the expression of PPARβat mRNA and protein levels (P<0.05).At the same time, the ex-pression of inducible NOS (iNOS) was obviously elevated (P<0.01), which occurred in parallel with the rising NOS ac-tivity and NO concentration (P<0.01).GW0742 (1 μmol/L), a selective PPARβagonist, inhibited the cardiomyocyte hypertrophy induced by HGI ( P<0.01 ) , and up-regulated the expression of PPARβat both mRNA and protein levels . Meanwhile, GW0742 also inhibited the increases in iNOS expression , NOS activity, and NO content induced by HGI , which were abolished by GSK0660 (1 μmol/L), a selective PPARβantagonist (P<0.01).CONCLUSION: PPARβdown-regulation and the following iNOS-NO activation are involved in the cardiomyocyte hypertrophy induced by HGI .

Objective To test whether glucagon like peptide-1 (GLP-1) would regulate the expression of visfatin in adipocytes,and to explore the mechanism of this effect.Methods Fully differentiated 3T3-L1 adipocytes were treated with GLP-1.Total RNA was extracted for analyzing the level of visfatin mRNA by quantitative RT-PCR.The media were collected for measuring the level of visfatin protein by enzyme linked immuno-assay (ELISA).In order to test the involvement of PKA pathway,the adipocytes were pretreated with a specific pharmacological PKA inhibitor H89 for 30 min before GLP-1 was added.Results GLP-1 increased visfatin expression in a time-and dose-dependent manner.The level of visfatin significantly increased at the concentration of 10 10 mol/L GLP-1 (P＜0.05),and reached the peak at 10-9 mol/L (P＜0.01).After incubation for 18 hours,GLP-1 dominantly increased the level of visfatin (P＜0.05).Inhibition of PKA pathway by H89 partially blocked the effect of GLP-1 on visfatin expression.Conclusions GLP-1 may enhance the expression of visfatin in 3T3-L1 adipocyte via the PKA pathway,which might contribute to the improvement in glucose homeostasis.

Objective To investigate the influence of severity of obstructive sleep apnea hypopnea syndrome (OSAHS)on acute ST -segment elevation myocardial infarction (STEMI).Methods 86 STEMI patients were divided into two groups,STEMI with OSAHS(OSAHS group,n=38)and STEMI without OSAHS(control group,n=48).Clinical data about biochemical index,cardiac function index and the duration of STEMI onset were compared between OSAHS group and control group.Logistic statistic analysis was used to investigate the risk factors that influ-ence the circadian rhythm of onset in STEMI.Results A total of 86 patients met the inclusion criteria,they were divided into two groups,STEMI with OSAHS(OSAHS group,n=38)and STEMI without OSAHS(control group,n=48).The incidence rate of STEMI onset during 0600 am~1 159 am was significantly higher in OSAHS group compared to control group(20.8% vs.44.7%,χ2 =5.626,P=0.018).This variation was weaken in mild OSAHS group compared to moderate-severe OSAHS group(20.8% vs.31.3%,χ2 =0.726,P=0.394;20.8% vs.54.5%,χ2 =7.956,P=0.005).Multivariate logistic analysis showed that the severity of OSAHS was a risk factor to the STEMI onset during 0600 am~1159 am(OR=2.458,95%CI 1.110~5.439,P=0.027).Conclusion The severity of OSAHS significantly increases the STEMI onset during 0600 am~1 159 am.

To obtain non-pathogenic rabies virus glycoprotein(RV-G),we expressed RV-G in Saccaromyces Cerevisiae(S.cerevisiae).In our study,tat-G fusion gene was cloned into the expression vector pYes2.0,which allows expression of a foreign gene in the yeast cells under the control of GAl1 promoter.Transforma-tion was performed by using lithium-treated yeast cells and several Ura+-tranformants were isolated.Ac-cording to the relative mobility in SDS-PAGE,we know probably two forms(designated as yGI and yGⅡ) of RV-G analogues produced in S.cerevisiae,their molecular weights were estimated as 66 kD and 56 kD,respectively.On the other hand,there was a specific band about 56 kD shown in western blot result.Com-bining precursors’ achievements,we will draw a conclusion that trans-membrane domain(TD) and cyto-plasmic domain have a negative regulation on RV-G antigen immunogenicity in S.cerevisiae.