Objective To explore the effect of embryonic stem cells(ESCs) differentiating into cardiomyocytes(CM) by cardiomyocytes.Methods The blastocytes of 3.5 d from Kunming mouse,cultured on mouse embryonic fibroblaste cell feeder layers for 4～5 d,the mouse ESCs were isolated and subsequently cultured.Day 2～3 embryoid bodies(EBs) were derived from ESCs of passage 3 to 5 and then incubated with neonatal cardiomyocyte(CM) to induce into cardiomyocytes and the expression of cardiac specific cardiac troponin-T(TnT) and ?-actin were detected by immunofluoresence.Results The rhythmic beating embryoid bodies were observed on day 3 and reached 93% of rhythmic beating embryoid bodies on 12 day in group 2.All the beating cardiomyocytes derived from ESCs expressed cardiac-specific proteins for TnT and ?-actin,and the group induced by direct cell contact acquired the highest differentiating ratio as 56.5%,and was the highest differentiating ratio compared with other groups(P

OBJECTIVE To investigate the positive rate of ?-lactamase,oprD2,aminoglycosides modifying enzyme and qacE△1 gene among clinical isolated strains of multi-resistant Pseudomonas aeruginosa in our hospital.METHODS P.aeruginosa was determined by VITEK,and MIC was determined by agar dilution method.TEM,IMP,VIM,oprD2,aac(3)-Ⅱ,aac(6′)-Ⅰ,aac(6″)-Ⅱ,ant(3″)-Ⅰ,ant(2″)Ⅰ and qacE△1 in strains were detected by polymerase chain reaction(PCR).RESULTS The positive rate of TEM,aac(3)Ⅱ,aac(6′)-Ⅱ,ant(3″)-Ⅰ and ant(2″)-Ⅰ in multi-resistant P.aeruginosa were 51.4%,48.6%,40.0%,54.3%,45.7% and 60.0%,respectively.No IMP and VIM genes were detected.CONCLUSIONS Positive rate of ?-lactamase, aminoglycosides modifying enzyme is high in 35 tested strains,and that should be paid more attention.

Objective To preparegraphene quantum dots (GQDs) and construct a novel biosensor for determination of dopamine (DA) and uric acid (UA).Methods The GQDs was prepared by carbonization method from citric acid as carbon sources.Differential pulse voltammetry was used by the modified electrodes to detect uric acid and dopamine,and the detection performance was evaluate.Collected three experimenters morning urine on July 29,2016.The proposed sensor was used for biological samples detection.Results The size of GQDs were between 3 to 5 nm,which was observed by transmission electron microscopy (TEM).The proposed sensor had good linear correlation of 50～600 μmol/L UA (r2 =0.996 6) and 2～ 240 μmol/L DA (r2 =0.992 5).In uric acid in urine samples' detection (n=3),RSD value was less than 3％.The standard addition recovery rates of UA and DA were in 95.7％～101.4％ and 96％～102％ respectively.Conclusion The biosensor based on GQDs for determination DA and UA had been constructed successfully.

Objective To investigate the effeets of ulimstatin on expression of intestinal defemin-5 mRNAin the rat model of sepsis.Method The experiment was performed in pharmaco-laboratory of medical college,Sun Yat-Sen University.sixty Sprague-Dawley rals were randomly divided into control,sepsis,pretreated andtreated groups(n=15).Semis was induced in the mts of latter three groups by cecal lifo.and puncture(CLP).The rats of pretreated group received 25 000 U/kg ulinastatin 2 hours before operation and the rats of uli-nastatin treated groups received 50 000 U/kg ulinastatin 2 hours after operation.Some pieces of ileum mucosa weretaken 12 h after CLP.Tge pathological changes were observed and the expression of RD-5 mRNA was detectedwith RT-PCR.All data were managed by SPSS 13.0 software and arIaIyzed by using One-way ANOVA and LSD-ttest.Results The expression of RD-5 mRNA in the rats of sepsis group significantly decreased compared to col-trol(P<0.05).The expression of RD-5 mRNA of pretreated and treated groups sigificantly inereased comparedto sepsis group(P<0.05);pretreated groups had more increased expression of,RD-5 mRNA compared to treatedgroups(P<0.05).Conclusions The expression of intestinal RD-5 mRNA significantly decreases in sepsis,which could be improved by the treatment of ulinadtatin leading to intestinal mucosal protection of the siqnifleant.The pretreatment may be more effective than the theTapeatic treatment in the rat model of sepsis.

Objective:To observe the changes of TLR4 and IL-6 expression in rats hippocampus CA1 region in remote ischemic postconditioning(RIP) and explore its significance.Methods: All the 72 male SD rats were divided into Sham group,Contrast group and RIP group randomly.Each group was divided into 4 time points:12h,24 h,48 h and 72 h group.There were 6 rats in each group.Use the cerebral ischemia-reperfusion model which was established with modified Longa method as the contrast group.The method of RIP was to Fasten rats Posterior limbs by a tourniquet for 30 min immediately,then relax them for 30 min, repeat 3 times.To observe the pathological variation of hippocampus CA1 region by HE dyeing;to test the expression of TLR4 and IL-6 by immunohistochemical staining.Results: Compared to contrast group, neuronal loss and swelling reduced significantly in RIP group.Compared to sham group, the TLR4 and IL-6 expression in contrast group and RIP group increased significantly ( P<0.05 ) .Compared to contrast group,the TLR4 and IL-6 expression in RIP group reduced significantly(P<0.05).Conclusion:RIP dose have protective effect on cerebral ischemia.The effect may be associated with the inhibition of TLR4 and IL-6 expression.

Aim: To compare the stability of Endostar~(TM) and endostatin under different temperatures and pH using polyacrylamide gel electrophoresis( PAGE) and Western blot and to compare the activity of Endostar kept at 4 ℃ and 37 ℃ by inhibition of endothelial cell proliferation. Methods: Endostar and endostatin expressed by Phicha pastoris were kept at 4 ℃, and 37 ℃ for 96 hours, respectively. The electrophoresis of the samples was detected by reduced and non-reduced PAGE. The results were further confirmed by Western blot with rabbit anti-Endostar polyclonal antibody. The inhibitory effect of Endostar stored at different temperatures on HUVEC was also exam-ined by cell-based assay. Results: Single band at 20 kD was detected in all lanes of SDS-PAGE gel loaded with endostatin and Endostar samples under reducing condition. In acidic native PAGE with three different pH values, endostatin showed a smear characteristic, whereas Endostar showed a unique band in acidic non-continuous native PAGE. Although the smear phenomenon was also observed under two conditions of constant native elec-trophoresis, the major band of Endostar could be detected. Similar electrophoretic behavior was found for endosta-tin and Endostar stored at both 37 ℃ and 4 ℃ . Western blot showed similar results to those by PAGE. Furthermore, Endostar stored at these two temperatures also had identical inhibitory effect on proliferation of HUVEC. Conclusion: Endostar and endostatin exhibit similar thermostability at regular conditions, but Endostar was more stable than endostatin expressed in P. pastoris under acidic condition.

Objective To evaluate the role of serine-threonine kinase (Akt)/glycogen synthase kinase-3 beta (GSK-3β) signaling pathway in isoflurane preconditioning-induced inhibition of mitochondrial permeability transition pore protein (mPTP) opening during myocardial ischemia-reperfusion (I/R) in rats.Methods Ninety-six male Sprague-Dawley rats,aged 3-4 months,weighing 200-250 g,were randomly divided into 4 groups (n =24 each) using a random number table:control group (group C);I/R group;isoflurane preconditioning group (group IPC);Akt inhibitor MK-2206 group (group MK).Myocardial I/R was induced by occlusion of the anterior descending branch of the left coronary artery for 30 min followed by 2 h of reperfusion.In group IPC,1.5％ isoflurane was inhaled for 30 min followed by 45 min washout,and then the model of myocardial I/R injury was established.In group MK,MK-2206 300 μg/kg (in dimethyl sulfoxide) was injected intraperitoneally at 30 min before isoflurane inhalation.At 2 h of reperfusion,8 rats were selected and sacrificed,and the hearts were removed for determination of myocardial infarct size.At 2 h of reperfusion,8 rats were selected,and blood samples were collected from the right internal jugular vein for determination of serum cardiac troponin Ⅰ (cTnI) concentrations.The rats were then sacrificed,and myocardial specimens were obtained for determination of the expression of phosphorylated GSK-3β (p-GSK-3β) in cytoplasm and mitochondria (by Western blot) and co-expression of p-GSK-3β with adenine nucleotide translocator (ANT),voltage-dependent anion channel or cyclophilin D in myocardial tissues (using co-immunoprecipitation).At 2 h of reperfusion,8 rats were selected and sacrificed,myocardial cells were obtained,and the opening time of mPTP was determined with a laser scanning confocal microscope.Results Compared with group C,the myocardial infarct size and serum cTnI concentrations were significantly increased,and the expression of p-GSK-3β in cytoplasm and mitochondria was up-regulated in I/R and IPC groups,the co-expression of p-GSK-3β with ANT was significantly down-regulated,and the opening time of mPTP was shortened in group I/R,and the co-expression of p-GSK-3β with ANT was significantly up-regulated,and the opening time of mPTP was prolonged in group IPC (P＜0.05).Compared with group I/R,the myocardial infarct size and serum cTnI concentrations were significantly decreased,the expression of p-GSK-3β in cytoplasm and mitochondria was up-regulated,the co-expression of p-GSK-3β with ANT was significantly up-regulated,and the opening time of mPTP was prolonged in group IPC,and the opening time of mPTP was significantly prolonged (P＜0.05),and no significant change was found in the other parameters in group MK (P＞0.05).Compared with group IPC,the myocardial infarct size and serum cTnI concentrations were significantly increased,the expression of p-GSK-3β in cytoplasm and mitochondria was up-regulated,the co-expression of p-GSK-3β with ANT was significantly down-regulated,and the opening time of mPTP was shortened in group MK (P＜0.05).No co-expression of p-GSK-3β with voltage-dependent anion channel or cyclophilin D was found in myocardial tissues.Conclusion The mechanism by which isoflurane preconditioning inhibits mPTP opening during myocardial ischemia-reperfusion is partially related to activation of Akt/GSK-3β signaling pathway in rats.

Objective To investigate the incidence and pregnancy outcomes of premature rupture of membranes (PROM) in pregnant women in Beijing.Methods A retrospective multicenter study of 18 534 cases delivered in Beijing Obstetrics and Gynecology Hospital,Beijing Friendship Hospital,Daxing MCH Hospital and Tongzhou MCH Hospital from January 2011 to December 2011,was conducted.Results Among 18 534 cases,PROM occurred in 4 504 cases (24.30％),including 3 910 cases of in term PROM (21.10％) and 594 cases of preterm PROM (3.20％).The incidence of premature delivery was 6.17％ (1 144/18 534),and among 1 144 cases of premature delivery 547 cases were PROM (47.81％);the incidence of PROM was 22.75％ (3 957/17 390) in term delivery.The overall cesarean section (CS) rate was 48.50％ (8 989/18 534) and that in pregnant women with PROM was 35.55％ (1 601/4 504),but the CS rate in pregnant women without PROM was 52.66％ (7 388/14 030).The rate of postpartum hemorrhage was 13.12％ (210/1 601)in CS cases and 4.17％ (121/2 903) in vaginal delivery cases (x2 =121.361,P=0.000).The mean hospital stay for PROM was (5.3±2.9) d in CS cases and (4.3±2.3) d in vaginal delivery cases (t =-12.136,P =0.000).Conclusions Without severe maternal or fetal complications,the incidence of PROM is relatively high in Beijing region and PROM may not increase the maternal or fetal complications.Vaginal delivery is the main mode of delivery for PROM.Cesarean section may not cause less neonatal complications,but may lead to more postpartum hemorrhage and longer hospital stay.

Objective To explore the prognostic risk factors of low level malignant obstructive jaundice treated with transhapetic biliary drainage.Methods The clinical data of 142 patients with low level malignant obstructive jaundice received percutaneous transhapetic biliary drainage management from January 2010 to June 2013 were retrospectively analyzed.The study parameters included gender,age,tumor type,preoperative obstructive time,preoperative infection,drainage method,Child-Pugh grade,serum total bilirubin (TBIL),albumin (ALB),serum creatinine (SCr),the postoperative declining degree of bilirubin and postoperative antineoplastic therapy.The prognostic risk factors were evaluated.Results Single variable analysis showed that preoperative infection (P =0.006),Child-Pugh grade (P =0.004),SCr (P =0.043),the postoperative declining degree of TBIL (P =0.001) and postoperative antineoplastic therapy (P =0.015) were the related factors for survival time.The further Logistic regression analysis showed that preoperative infection (OR =3.729,95％ CI 1.332-6.363,P =0.040),Child-Pugh grade ≥ 10 scores (OR =0.513,95％ CI 0.375-1.276,P =0.018) and postoperative antineoplastic therapy (OR =0.668,95％ CI 0.210-2.026,P =0.038) were the related factors for survival time.Conclusion In treating of low level malignant obstructive jaundice with transhapetic biliary drainage,the preoperative infection,Child-Pugh grade and postoperative antineoplastic therapy may be the important related factors that affect the patient's survival time,to evaluate the prognosis of these patients has important reference meaning.

BACKGROUND: Hundreds of mouse embryonic stem cell lines are established presently. Most of these cell lines are collected from 129, C57BL/6J and BALB/C mice. Success rate of creating embryonic stem cell lines with Kunming mice is very low. OBJECTIVE: To explore the method of isolation and culture of murine embryonic stem cells of kunming species to increase the efficiency of establishment of embryonic stem cell lines from Kunming species mouse. DESIGN, TIME AND SETTING: The cell experiment was performed at the Chongqing Key Laboratory of Neurology from April 2006 to June 2007. MATERIALS: Blastocysts of 3.5 days and 4 days from Kunming species mouse were collected. METHODS: The blastocysts of 3.5 days and 4 days from Kunming species mouse were cultured on the fibroblast cell feeder layers for 4-5 days, and the cells from inner cell mass were isolated and subsequently cultured in vitro in 2.5 g/L trypase-0.4 g/L ethylenediamine tetraacetic acid (EDTA) and 1 g/L trypase-0.2 g/L EDTA at room temperature for 2-4 minutes. Embryonic stem cell colony was mechanically straggled. MAIN OUTCOME MEASURES: Colony growth was observed and determined by alkaline phosphatase staining, OCT-4 staining and karyotype analysis. RESULTS: Adherence rate, inner cell mass frequency and cloning efficiency of 4 day post coitum (dpc) embryo were higher than those of 3.5 dpc embryo (P