Objective To observe the injury to the heart,liver and kidney induced by hind limb ischemia/reperfusion(I/R),and to study the protective effect of H2S by attenuating this injury,in order to offer a theoretical basis for therapeutic application.Methods A model of hind limb I/R was reproduced.28 rats were divided randomly into 4 groups:normal control group(C,n=8),ischemia 4h group(I,n=8),ischemia 4h and reperfusion 4h group(I/R,n=6)and I/R+NaHS group(n=6)(NaHS can provide H2S for rats in vivo).The concentrations of CK,CK-MB,Tn-T,ALT,AST,Cr and urea in blood plasma were determined.The pathological changes in the heart,liver and kidney were observed with HE staining.Results The concentration of CK,CK-MB and Tn-T in I group increased significantly compared with that in C group,and that in I/R group was higher than in I group.No significant difference was found in plasma concentration of ALT,AST,Cr and Ur between C group and I group,while it they were increased significantly in I/R group compared with that in I group,and that in I/R+NaHS group were decreased significantly compared with that in I/R group.Pathological examination revealed pathological findings of injury to the heart,liver and kidney in I group,and even more serious injuries were found in I/R group than in I group,while the they were less marked in IR+NaHS group compared with that in I/R group.Conclusion Hind limb ischemia/reperfusion may result in injury to the heart,liver and kidney.H2S may attenuate injury to remote organs follawing hind limb ischemia/reperfusion.

Objective To investigate the effect of meisoindigo inducing primary K562 cell apoptosis and its influence on the expression of conduction signal STAT5.Methods K562 cell was treated with meisoindigo for 12 h、24 h、48 h respectively.Flow cytometry was used to detect cell apoptosis and cell cycle,and Western blot was employed to examine the alteration of the expression of STAT5.Results ①The rate of K562 cell apoptosis treated with meisoindigo for 12 h、24 h、and 48 h was 12.5 ± 0.69,19.4± 1.07 and 42.5 ± 3.22,showing statistical significance compared with the control group (P＜0.05).②Meisoindigo could block K562 cell at the S stage,the ratio of S stage cell was increased to 70.48±6.34 with the decrease of cells at G0/G1 and G2/M stage.③Gradually reduced expression of STAT5 showed after K562 cells treated with meisoindigo from 12h to 48h.Conclusion Meisoindigo accelerrates the apoptosis of K562 cell and the mechanism may relevant to the decreased expression of STAT5.

Objective To investigate factors and neonatal outcomes associated with histologic chorioamnionitis (HCA) in preterm premature rupture of membranes (PPROM).MethodsFrom Jan.2008 to Jun.2011,230 women with PPROM at 28 -33 +6 weeks of gestation undergoing deliveries in the Second Affiliated Hospital of Wenzhou Medical College were studied retrospectively.According to placental histopathologic findings,those patients were categorized into two groups,including 138 cases in histologic chorioamnionitis (HCA group ) and 65 cases in non-chorioamnionitis (control)group.Age,parity,gestational age of PPROM and delivery,latency period,oligohydramnios,white blood cell (WBC) count and serum C-reactive protein (CRP) level at admission and before delivery,the incidence of neonatal respiratory distress syndrome (NRDS),neonatal pneumonia,bronchopulmonary dysplasia,necrotizing enterocolitis,early-onset neonatal sepsis,abnormal brain sonography findings and mortality were compared between two groups.Results( 1 ) The incidence of HCA was 68.0.％ ( 138/203 ) in all 203 cases with PPROM.(2) The occurring ruptured membrane gestation in HCA group was ( 31.1 ± 1.5 ) weeks,which were significantly earlier than (32.0 ± 1.3 ) weeks in control group ( P ＜ 0.05 ).The level of CRP of (8.2 ± 14.9) mg/L before delivery in HCA group was significantly higher than (5.5 ±7.2) mg/L in control group (P ＜ 0.05).The rate of oligohydramnios and cesearean sections were 55.1％ (76/138) and 45.7％ (63/138) in HCA group,which were significantly higher than 30.8％ (20/65) and 29.2％ (19/65) in control group (P ＜0.05).There were no significant difference in patient's age,parity,WBC count and CRP at admission between two groups (P ＞ 0.05 ).The latency period did not show significant difference between (140± 116) hours in HCA group and (129 ± 125) hours in control group (P ＞ 0.05).(3) Using multivariable logistic regression models,oligohydramnios ( OR =2.937 ),gestational age of PPROM ＜ 32 weeks ( OR =2.352),serum CRP level ＞ 8 mg/L before delivery ( OR =4.923 ) and latency period ＞ 48 -168 hours (OR =4.439) were significantly associated with HCA (P ＜0.05).(4) The gestational age of delivery and birth weight of HCA group were significantly lower than those of control group [ ( 32.0 ± 1.5 ) weeks vs.( 32.7 ± 1.5 ) weeks,( 1680 ± 379) g vs.(2017 ± 333) g,respectively,P ＜ 0.05 ].The incidence of Apgar ＜7,abnormal brain sonograhy findings, neonatal pneumonia,bronchopulmonary dysplasia,early-onset neonatal sepsis and mortality in HCA group were significantly higher than those in control group [20.3％ (28/138) vs.7.7％ (5/65),14.5％ (20/138) vs.4.6％ (3/65),12.3％ (17/138) vs.3.1％(2/65),5.8％ (8/138) vs.0,6.5％ (9/138) vs.0,12.3％ (17/138) vs.3.1％ (2/65),respectively,P ＜ 0.05 ].The incidence of necrotizing enterocolitis ( 1.5％,2/138 ) in HCA group was higher than that of controlgroup(0) and the incidence of NRDS ( 18.8％,26/138) in HCA group did not show statistical difference with 21.4％ ( 14/65 ) in control group ( P ＞ 0.05 ).ConclusionsIt was found that HCA was significantly correlated with lower gestational age of PPROM,higher serum CRP level before delivery,prolonged latency period and oligohydramnios in PPROM.HCA could increase the neonatal morbidity and mortality.

Medical disputes are long-existing and tough social contradictions.In recent years,the third-party mediation is tried out and proved to be an innovative solution to medical disputes.This paper takes Jining medical rights safeguard association as an example to darify the significance of the third-party mediation in handling medical disputes and safeguarding legal rights of patients.

OBJECTIVETo investigate the optimal conditions of tri-expression of CYP3A4, POR and cyt b5 in Sf 9 cells.

METHODSThe Sf 9 cells expressing CYP3A4, POR and cyt b5 were cultured in shaker flasks. The optimized conditions, including the temperature and rotation speed, the culture volume, the amount of surfactant and the culture time were studied. The expressed products in microsomes were used to metabolize the testosterone and their metabolic activity was determined.

RESULTSWhen the temperature and rotation speed of the shaker were 27 degree and 90 r/min, the cell density and culture volume were 5X105 cells/ml and 80-120 ml per 250 ml shaker flasks, respectively. When Pluronic F-68 was 0.1% and the culture time was 72 h, the condition was most suitable for culture of Sf 9 cells and expression of targeted proteins. When the ratio of the volume of three added viruses was 1:1:1, the expression condition was optimal, under which the Km, Vmax, and CLint for testosterone metabolism were 119.6 μmol/L,0.52 μmol/(min*g protein) and 4.34 ml/(min*g protein), respectively.

CONCLUSIONThe conditions of tri-expressing of CYP3A4, POR and cyt b5 have been optimized in the study and the product CYP3A4 is obtained with higher metabolic activity.

Animals ; Cytochrome P-450 CYP3A ; biosynthesis ; Cytochromes b5 ; biosynthesis ; Humans ; Insecta ; NADPH-Ferrihemoprotein Reductase ; biosynthesis ; Sf9 Cells

Objective To evaluate the characteristics of the PREVI Isola automated plate streaker (bioMérieux,SA).Methods 80 respiratory tract specimens,70 sterile fluids,52 stools,69 swabs,12 cerebrospinal fluids and 80 urines were collected in Peking Union Medicd College Hospital.Specimens were processed with manual streaking and PREVI Isola system.PREVI Isola system were evaluated comparing to the manual streaking.The quality of results were analyzed by SPSS 16.0, doing Wilcoxon’s Sign Rank Test for the results of finally isolated species,overall numbers of isolated colonies and semi-quantita-tive of the species were both isolated by the two methods.Results PREVI Isola system was highly automatic,which could select the right plates and stake the bar code on the back of the plate indicating the type of the agar and inoculation time, PREVI Isola system could accurately absorb the liquid specimen and use a novel comb streaking procedure for processing of fluid specimens on standard agar plates,like 17 inoculating loops work together.It also had a good reproducibility.The quali-ty of PREVI Isola system results:As to the finally isolated species,there was significant statistical difference between PREVI Isola system and manual streaking method in respiratory tract and stools specimen,there were more species isolated by manual streaking method than PREVI Isola system.There were no differences between the two methods for the other of specimen types.As to the amount of pure clones of the species were both isolated by the two methods,there were significant statistical differences between the two methods for respiratory tract,sterile fluid and stool specimens.The amount of clones isolated by PREVI Isola system was more than manual streaking method.In semi-quantitative results,there were significant statistical differences between the two methods for respiratory tract and urine specimen,Species had wider distribution of PREVI Isola system than manual streaking method.Inoculation efficiency:if the batch of specimen type was simple (mainly the urine and so on),using the same plates,PREVI Isola system was more efficient than manual streaking method.Howev-er,if the batch of specimen type was complicated,manual method was high-performance.Besides,not all specimen type could be inoculated by PREVI Isola system,such as cerebrospinal fluid,catheter and tissues.Conclusion If the lab had simple specimen type,or utilize the specimen type using the same agar plates to be inoculated together,PREVI Isola system belongs to a good performance automated plate streaker.

Objectives To evaluate the application of preoperative endoscopic ultrasonography ( EUS) in assessment of invasive risk and selection of therapeutic modalities for gastric stromal tumors ( GST).Methods The clinical data of 135 patients with GST admitted in our hospital from January 2011 to January 2012 were retrospectively analyzed.The invasion extent of GST was assessed by image of EUS before surgery, and compared with pathological results after surgery; the Fletcher 4-tier system was used for predicting the aggressiveness of GST.The selection of therapeutic modalities in 38 patients, who underwent surgical treatment was analyzed.Results No specific clinical manifestations were noticed , but some patients with enormous GST had symptoms of ulcer , hematemesis , and melena.Among 135 patients 97 cases received conservative treatment and followed up;in remaining 38 cases, according to invasion risk assessed by EUS, there were 9 cases in low risk, 18 in intermediate risk and 11 in high risk.The surgical modalities were selected based on the risk assessment:endoscopic therapy was performed in 15 cases, laparoscopic with gastroscopic surgery in 17 cases and laparotomy in 6 cases.The coincidence rate of diagnosis between preoperative EUS and postoperative pathological examination was 79.0%.Conclusions Preoperative endoscopic ultrasonography is of value in assessment of invasion risk and selection of appropriate therapeutic modalities for gastric stromal tumors.

OBJECTIVE To investigate predisposing factors of candidemia in nosocomial infections.METHODS To retrospectively review the clinical features of 120 cases,including 59 who developed candidemia and 61 cases with bacteremia during the period of 1990-2004.RESULTS The incidence of candidemia was stable over a 14-year period.Candida albicans remained the predominant Candida species recovered(30.5%),followed by C.tropicalis(25.7%),C.glabrata(12.9%),C.parapsilosis(12.9%) and others(17.1%).Of the total 59 cases of candidemia,were administrated by broad spectrum antibiotic therapy for long time,urinary catheters,malignant tumor,etc.Multivariate analysis showed that candidemia was related with many factors.CONCLUSIONS C.albicans was the major pathogen in our hospital during 14 years;the candidemia is related with the use of quinolones,ventilator,central venous catheters and radiation-chemotherapy(P

Objective To compare the detecting results of rotavirus (RV)and adenovirus (AdV)antigens using auto stool pretreatment system (machine method)and manual method.Methods A total of 100 stools collectecd from diarrear patients admitted in gastroenterology outpatient department from September 2014 to Octorber 2014 in Peking University Medical College Hospital were detected to identify RV and AdV antigens using machine method and manual method respectively,and the nucletic acids of positive samples were detected by liquid chip method to verify the results.Results The RV,AdV and co-infection antigen positive detection rate using machine method were 17.0% (17/100),25.0% (25/100)and 12.0% (12/100)respectively,whereas those using the manual method were 4.0% (4/100),13.0% (13/100)and 2.0% (2/100),re-spectively.Taking the nucletic acids detection as the golden method,the false positive detection rate of RV,AdV and co-in-fection antigen using machine method were 23.5% (4/17),20.0% (5/25)and 33.3% (4/12)respectively,whereas those u-sing the manual method were 75.0% (3/4),69.2% (9/13)and 50.0% (1/2),respectively.χ2 test for paired data for RV and AdV positive detection rate,false positive detection rate of RV and false positive detection rate of AdV using two meth-ods were statistically significant (χ2 =15.0,52.8 and 47.5,P values <0.05).Two methods for detecting RV and AdV had poor consistency (Kappa value was 0.25,Kappa values <0.4).Conclusion Machine method has much more advantage on RV and AdV positive detection rate and false positive detection rate than manual method,which is good for clinical applica-tion.

Objective To evaluate the in vitro activity of daptomycin, vancomycin, teicoplanin, tigecycline, ceftobiprole and linezolid against 499 strains of blood-isolated gram-positive cocci. Methods Determination of the minimal inhibitory concentration (MICs) of daptomycin with microbrothdilution method and the MICs of other 9 antimicrnhial agents with agar dilution method against 499 strains of blood-isolated gram positive cocci was carried out. The data was analyzed with WHONET 5.4 software. Results The susceptibility rates of staphylococci to daptomycin, tigecycline, linezolid, ceftobiprole, vancomyein and teicoplanin were 100%. All staphylococcus strains were inhibited by daptomycin at a MIC of 1 mg/L. The MIC50 and MIC60 of daptomyein were both 0.5 mg/L against methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-resistant Staphylococcus coagalase-negative (MRSCnN). Among Enterococcus spp, the highest MIC of daptomycin was 4 mg/L. The MIC50 and MIC90 of daptomycin were both 2 mg/L against E.faecalis, whereas they were 2 mg/L and 4 mg/L against E.faecium. One strains of linezolid-resistant E.faecalis(MIC:8 mg/L)was susceptible to daptomycin (MIC: 1 mg/L). Three strains of E.faecium carrying vanA gene with vancomycin MICs above 32 mg/L and teicoplanin MICs also 32 mg/L were susceptible to daptomycin, tigeeycline and linezolid. The MIC range of daptomycin against Streptococcus pneumoniae and Streptococcus viridans was 0.032-0.25 mg/L and 0.125-1.000 mg/L separately. Conclusions Daptomycin has excellent in vitro activity against common gram-positive pathogens isolated from blood. It may be a good choice for clinicians to treat drug-resistant gram-positive cocci.