Objective To detect of clostridium perfringens by qPCR in mouse models and a clinical case in order to offer early diagnosis.Methods 40 Kunming mice were randomly grouped and intramuscular injected clostridium perfringens type A in leg 0.1 ml(3.5 × 109cfu/ml or 3.5 × 108cfu/ml or 3.5× 107cfu/ml,diluted with saline),while control group was injected with 9% sodium chloride 0.1ml.The mouse models and a clinical case were detected by qPCR.Results The death rate of 3.5 × 109,3.5 × 108,3.5 × 107cfu/ml and the blank group were 90%,70%,10% and 0% after intramuscular injection for 72 h spectively.The mean Ct values among these groups were 21.21 ±2.69,28.45 ±2.74,32.49 ±2.87 and 0.00 ± 0.00(P ＜ 0.05).The Ct values of the patient were 30.67 and 30.44.Conclusions Cclostridium perfringens could be successful identified with qPCR in mouse models when the mice still did not show any symptoms.

Objective To evaluate the clinical application of computer-aided electromagnetic imaging navigation in nasal endoscopic surgery.Methods Twenty-two cases of nasal endoscopic surgery with intraoperative imaging navigation were retrospectively reviewed,including 16 cases of sinusitis with or without polyp;5 cases of nasal inverting papilloma; 1 case of maxillary capillary hemangioma.All cases were operated with computer-aided electromagentic imaging navigation and nasal endoscope.Results The preoperative preparing time would take 4-10 minutes.In 22 cases,the localization accuracy between 3-D image landmarks of navigation system and actual anatomical landmarks was less than 1 mm.The optic nerve and other anatomical landmarks could be orientated accurately in intraoperative procedures.No complication occurred.Conclusions Nasal endoscope combined with computer-aided electromagnetic imaging navigation provides accurate anatomical localization of nasal cavity,sinuses and anterior skull base.It could improve the effectiveness and decrease surgical complications,especially in complicated cases.

Objective To study the changes of coagulation function and platelet-related parameters of patients with acute pancre-atitis(AP) and their clinical significance .Methods 36 patients with AP were served as the AP group ,38 healthy people ,the control group .Healthy people in the control group and patients in the AP group at the time of admission and remission were subjected to detection of serum amylase ,lipase ,leukocytes ,neutrophils ,lymphocytes ,platelets ,mean platelet volume(MPV) ,platelet distribution width(PDW),prothrombin time(PT),thrombin time(TT),activated partial thromboplastin time(APTT),prothrombin activity (PTA) ,international normalized ratio (INR) ,fibrinogen(FIB) and D-dimer(D-D) levels .Results Levels of serum amylase ,lipase , TT ,INR ,FIB ,and D-D of patients in AP group were significantly higher than those in the control group(P<0 .01) ,while their PT , PTA levels were significantly lower than those in the control group (P<0 .01) .Differences of leukocytes ,neutrophils ,lymphocytes and MPV of AP patients between at the time of admission and remission were statistically significant (P<0 .01) .Conclusion De-tections of coagulation function and platelet-related parameters changes of AP patients contribute to evaluation of the patients′con-dition and prognosis .

Objective:To investigation of the long-term-siRNA treatment with HBV transgene mice on inhibit replication of hepatitis B virus .Methods:The constructed siRNA expressed vectors was transfected HBV transgene mice by hydrodynamics -based in-jection via vena caudalis .Different groups were set including:specificity siRNA groups ( pSilencer5.1/C2,pSilencer4.1/C2,pSilenc-er3.1/C2),PBS group and negative vector group (n=10).The effect was observed in different periods (6 d,21 d,1 months,3 months, 6 months and 9 months after injection ) .HBsAg was analyzed by Chemiluminescence method , HBV-DNA was analyzed by real time quantitative PCR ( RQ-PCR) .Results:Compared with the PBS group , specificity siRNA groups showed decreased levels of HBsAg and HBV-DNA (P<0.05).Negative vector group did not show such changes ,there were no significant differences (P>0.05).Conclu-sion:The siRNA based on the expression vector can suppress the expression and replication of HBV in HBV transgene mice .The inhi-bition effects of long-term-siRNA treatment was specific .

Objective Through observing and treating the ovulation dysfunction patients with birth demand ,to study the clinic characteristics and therapy strategy .Methods 630 clinical cases including natural cycles and controlled ovarian stimulated cycles . monitored by transvaginal B-ultrasonography from April 2008 to April 2012 ,The common reasons ,clinical manifestation ,and out-come undergoing different treatment strategies were analyzed .Results In the natural cycles ,41 .61% patients suffered ovulation dysfunction ,PCOS patients occupied the most .Through the therapy of controlled ovarian stimulating on these patients ,60 .84% of ovulation dysfunction patients recovered normal ovulation .The therapeutic regimen of clomiphene citrate (CC) 50 mg and of CC combined with human menopausal gonadotropin(HMG) showed a higher ovulation rate ,66 .49% and 67 .57% respectively(P<0 .05) .Anovulia was the most commonly type of the ovulation dysfunction ,followed small follicle ovulation and luteinizing unrup-tured follicle .Conclusion Ovulation dysfunction is frequent in infertility patients .Understanding the clinical characteristic and the disease cause ,working out the favourable and effective therapeutic regimen can increase their conception possibility .

Following the wishes of volunteer patients,316 cases of type 2 diabetics of our community center in 2008 carried out a three-year intervention from 2008 to 2011 on the basis of rational drug therapy plus the interventions of health education,regular review and lifestyle strengthening.And regular monitoring and recording were performed on the parameters of body mass index (BMI),blood pressure,blood lipids,fasting glucose,2 h postprandial blood glucose,glycosylated hemoglobin (HbA1c) and liver & kidney function.After intervention,some indicators changed:BMI (25.2 ±3.5) vs.(25.0 ±3.3) kg/m2,systolic blood pressure (129.1 ± 11.8) vs.(126.2 ±7.9) mm Hg(1 mm Hg=0.133 kPa),fasting glucose (7.80 ±2.81) vs.(7.25 ± 1.96) mmol/L,2 h postprandial blood glucose (11.04 ±4.60) vs.(9.83 ±3.60) mmol/L,HbA1c (7.39 ± 1.61) vs.(7.17 ± 1.65)％,total cholesterol (5.08 ±1.21) vs.(4.74 ± 1.35) mmol/L,low density lipoprotein cholesterol(LDL) (3.09 ± 0.87) vs.(2.85 ±0.83) mmol/L,high density lipoprotein cholesterol (HDL) (1.27 ± 0.33) vs.(1.41 ± 0.32) mmol/L,serum creatinine 65 vs.72 μmol/L,uric acid 300 vs.317 μmol/L.And the differences were statistically significant (P ＜ 0.05).After intervention,blood pressure compliance rate increased from 72.5％ to 88.0％,LDL-C compliance rate improved from 27.2％ to 38.6％,HDL-C compliance rate of 54.9％ increased to 66.9％.And the differences were statistically significant (P ＜ 0.05).The pre-intervention combined compliance rate of 11.4％ (n =36) rose to 17.7％ (n =56).And there was significant difference (P =0.024).

Objective To investigate the effects of 1,25-dihydroxyvitamin D3[1,25 (OH)2D3] on cell proliferation in hu?man glomerular mesangial cells and it′s effects on the regulation of mTOR/p70s6K signaling pathway in this cell line. Meth?ods The cultured human mesangial cells at passage 3-7 were divided into four groups:control group,VD group (addition of 10-8 mol/L of 1,25-dihydroxyvitamin D3 ),R group (addition of 5 mg/L of rapamycin) and R+VD group(addition of 5 mg/L ra?pamycin combined with 10-8 mol/L of 1,25-dihydroxyvitamin D3). Drug incubation last 48 h. The effect of mesangial cell pro?liferation was measured by CCK-8 colorimetric assay. The cell cycles were measured by flow cytometry. The expression of mTOR and p70s6K were detected by immunofluorescence. Results (1) The absorbance of A450 was higher in control group than that in VD group than that in R group than that in R+VD group. But the inhibition rate (IR) was lower in control group than that in VD group than that in R group than that in R+VD group. All comparisons were of statistic significance. ( 2) Cells in G1 phase were higher while cells in G2/M and S phases as well as proliferation rate (PI) were lower in control group than those in VD group than those in R group than those in R+VD group. All comparisons were of statistic significance except in?dexes between group R and group VD. (3) mTOR and p30s6K expressions in mesangial cells were higher in control group than those in VD group than those in R group than those in R+VD group. All comparisons were of statistic significance ex?cept indexes between group R and group VD. Conclusion 1,25-dihydroxyvitamin D3 might inhibit mesangial cell prolifera?tion significantly through mTOR/p70s6K signaling pathways.

Objective To explore the effect and the possible pathway of different concentrations of QLT0267,which was the inhibitor of the integrin-linked kinase (ILK),on the process of high glucose-induced tubularepithelial-myofibroblast transdifferentiation (TEMT) in human renal tubular epithelial cells (HK-2).Methods HK-2 cells were exposed to 30 mmol/L GS,and TEMT model was established.After excluding the effect of high osmotic in TEMT,HK-2 cells were divided into 6 groups by different concentrations of GS and QLT0267 for 48 hours.The rate of the cell proliferation was calculated by MTT.The expression of ILK and α-smooth muscle actin (α-SMA) were determined by immunofluorescence and Western blot,and the expression of protein kinase B (AKT),phosphorylated protein kinase B (p-AKT),and E-cadherin were determined by Western blot.Results (1) The expression of ILK,p-AKT,and α-SMA in HK-2 cells were unregulated and the expression of E-cadherin was downregulated for 48 hours with glucose treating vs control (P ＜ 0.05);(2) The proliferation rate in high glucose group was higher than the group which concentration of QLT0267 was greater than 5 μmol/L (P ＜ 0.05);(3) With the concentrations of QLT0267 increased,the expression of p-AKT,α-SMA was gradually decreased (all P ＜ 0.05),and the expression of E-cadherin was gradually increased (all P ＜ 0.05).Conclusions 30 μmol/L of GS can lead to TEMT in HK-2 cell.The QLT0267 with concentration greater than 5 μmol/L may prevent the activation of ILK downstream proteins,then partially inhibits cell proliferation and TEMT in HK-2 cell.

Objective To study the nutritional risks in elderly patients with prostate cancer and explore its related factors.Methods 346 elderly patients with prostate cancer in Wuhan area from May 2013 to May 2014 were chosen as the objects in this study.Nutritional risk screening 2002 was used to evaluate nutritional risk.The sleep quality,anxiety,lower urinary tract symptoms,differentiation and other data of patients were collected.The influencing factors for nutritional risk in the patients were analyzed.Results The ratio of nutritional risk in elderly advanced prostate cancer patients was 35.3％ (122/346).The nutritional risk was increased with age (P＜0.05).The prostate cancer patients with nutritional risk had a higher international prostate symptom score (IPSS) (P＜ 0.05).Monovariate factor analysis showed that sleep quality,operation,pathology staging,radiotherapy,chemotherapy,lower urinary tract symptoms (LUTS) were the impact factors for nutritional status in elderly patients with prostate cancer (P＜0.05).Logistic regression analysis showed that age (OR=0.29),sleep quality (OR=0.25) were the protective factors for the nutritional status,while surgery (OR=12.67),pathological staging (OR=1.65),radiotherapy (OR=2.65),SPSS (OR=1.55),chemotherapy (OR=1.85) were the risk factors for nutritional status (P＜ 0.05).Conclusions The incidence of nutritional risk is high in elderly prostate cancer patients.Age,sleep quality are the protective factors,and operation,pathology staging,SPSS,chemotherapy,and radiotherapy are the risk factors for nutritional status.

Objective To investigate the clinical significance of testing serum kisspeptin in central precocious puberty (CPP) girls. Methods Sixty eight CPP girls and 68 healthy girls was studied from December 2012 to December 2014. HEK293 cells were cultured. Luciferase reporter assay was performed to verify the binding of miR-137 to the 3′UTR of KISS1. Serum miR-137 level was levaluated by qRT-PCR. Level of serum luteinizing hormone , prolactin , follicle stimulating hormone , thyrotropin , free thyroxine and estradiol was evaluated by chemi-luminescence immunoassay. The level of serum kisspeptin was detected by ELISA. Results MiR-137 was confirmed to bind to the 3′UTR of KISS1. The level of serum miR-137 was downregulated and kisspeptin was enhanced in CPP girls. The expression of miR-137 and kisspeptin was negatively correlated. Serum miR-137 level was negatively related to bone age and bone age advancement. According to the results of GnRH stimulating test, serum miR-137 was related to peak LH and peak/basal LH ratio. Conclusions MiR-137 could bind to the 3′UTR of KISS1. MiR-137 may be a potential biomarker for CPP assisted diagnosis.