Objective Ouabain is a cardiotonic steroid that can induce the apoptosis of many tumorous cells .This study was to investigate the anti-tumor mechanisms of ouabain by observing its effects on the apoptosis of T lymphoblastic leukemia Jurkat cells and the expressions of hTERT and c-myc mRNA and protein . Methods Jurkat cells were treated with ouabain at the concentrations of 50 and 100 nmol/L for 24 and 48 hours, and those treated with 1 ×PBS served as the control .Then the apoptosis rate of the cells was detected by flow cytometry after Annexin V/PI staining, the expressions of hTERT and c-myc mRNA determined by RT-PCR, and those of hTERT and c-myc protein by Western blot . Results The apoptosis rates of the Jurkat cells in the 50 and 100 nmol/L oua-bain groups were (5.67 ±3.71)%and (9.63 ±4.83)%respectively at 24 hours, and (19.67 ±4.55)%and (37.60 ±11.89)%at 48 hours, significantly higher than (4.23 ±1.01)%in the PBS control group at 48 hours (P<0.05).Compared with the control, the expressions of hTERT and c-myc mRNA were decreased by 200%and those of hTERT and c-myc protein by 224%and 400%, re-spectively, at 48 hours (P<0.05).There was a positive correlation between the reduction of the mRNA levels and that of the protein levels of hTERT and c-myc (P<0.05). Conclusion Ouabain can down-regulate the mRNA and protein expressions of hTERT and c-myc, which may be one of the mechanisms of its induction of the apoptosis of Jurkat T lymphocyte leukemia cells .

Objective To discuss the clinical character,mechanism and prevention of cerebral-cardiac syn-drome(CCS) during acute stroke. Methods 189 patients with CCS caused by acute stroke were Analysed retrospec-tively. Results The incidence rate of CCS following stroke was 55.0%. 84.6% patients with CCS occur in the first 72h. The dead rate of acute stroke with CCS was 26.9%, significartly higher than that of without CCS( P<0.01 ). Conclusion CCS is associated with type and pathologic site of acute stroke. The prognosis of acute stroke with CCS was worse than that without CCS.

Objective Discoidin domain receptor 2 is a kind of receptor tyrosine kinases, which was found to be over-expressed in synovial fibroblasts in rheumatoid arthritis (RA). The present study was to construct and express wild type (FLDDR2), Fc chimera (FcDDR2) and truncated form of mouse discoidin domain receptor 2 (ttDDR2) for further study. Methods Full-length, truncated form of mouse DDR2 were amplified by RT-PCR. A chimera form of mouse DDR2 was constructed by replacing part of the extracellular domain with Fc fragment of human immunoglobulin G1 (IgG1). Eukaryotic expression vectors of the different forms of mouse DDR2 were constructed by subcloning the PCR products into pcDNA3.1(+) or pMKIT-Neo. Then COS-7 cells were transient transfected with the eukaryotic expression vectors of full-length (FLDDR2), truncated (ttDDR2) and chimeric form of mouse DDR2 (FcDDR2). Successful transfection and expression was confirmed by Western blot(WB) and Immunoprecipitation (IP)/WB. With or without stimulation with soluble type I collagen for 3h, phosphorylation level of transfected cells were detected by IP/WB. Eukaryotic expression vectors of full-length, truncated form and chimeric forms of mouse DDR2 were successfully constructed and confirmed by sequencing. After transient transfection, the expression of these three forms in the respectively transfected cells was observed by Western blot. Results The result of IP/WB suggested that the chimeric form of mouse DDR2(FcDDR2) could be properly expressed in the COS-7 cells. Under the condition of collagen, decreased tyrosine phosphorylation of FLDDR2 was detected in the COS-7 cells that were cotransfected with ttDDR2 and FLDDR2, comparing with that in COS-7 cells transected with only FLDDR2. There was no obvious difference in phosphorylation level between FcDDR2 without collagen stimulation and FLDDR2 with the collagen stimulation. Conclusion Three different forms of mouse DDR2 were successfully constructed. FcDDR2 could be an activator for its auto-phosphorylation. And ttDDR2 could be a partially negative competitor.

Aim To investigate total flavones of epimedium (TFE) modulation of glycine-activated current (IGly) in acutely dissociated rat spinal dorsal horn neurons. Method Whole-cell patch-clamp technique was used. Result TFE inhibited IGly reversibly in a concentration-dependent manner. It shifted the concentration-response relationship to the right without altering the maximum response and Hill coefficient of the IGly. TFE did not change the ion selectivity of glycine receptor either. Conclusion These results indicate that TFE directly inhibited the glycine receptor response by decreasing the affinity of glycine to its receptor.

Objective To evaluate the color Doppler flow imaging (CDFI) findings in predicting possible difficulties of laparoscopic cholecystectomy (LC). Methods A total of 99 patients with acute cholecystitis received CDFI examination, and were divided into easy LC group (n=67) and difficult LC group (n=32) according to the indexes including the volume of gallbladder, the thickness of gallbladder wall, the condition of arterial flow in the gallbladder wall, the condition of gallbladder cavity and fossa, and of the intra-and extra-hepatic bile duct. After LC, all the patients were redivided into easy LC group and difficult LC group according to a difficulty scoring system of LC. The relationship among imaging results, operation difficulties and operation findings was investigated. Results According to the final score of difficulty scoring system of LC, the number of patients in easy LC group and difficult group was 61 and 38, respectively. Compared with easy LC group, patients in difficult LC group had bigger gallbladder volume [(39.5±13.2)am3 vs(32.6±10.4)cm3], thicker gallbladder wall [(10.1±4.0)mm vs(3.8±0.9)mm], more stone incarceration in the neck of gallbladder, abundant color blood flow signal of the gallbladder wall and more severe adhesion of gallbladder to the adjacent tissues, with statistical difference between the 2 groups (t=-2.820,-12.318,-3.952,x2=33.548,19.461,P<0.05). Using gallbladder volume, the thickness of gallbladder wall, infarction of stones in the neck of gallbladder and adhesion of gallbladder to the adjacent tissues as prediction indexes, the accuracy of CDFI in predicting the possible difficulties of LC was 94% (93/99). Conclusion Preoperative CDFI is helpful in predicting difficulties of LC for acute cholecystitis.

Objective To investigate the feasibility,safety and efficacy of laparoscopic and endoscopic cooperative surgery (LECS) for elderly patients with gastric gastrointestinal stromal tumor (GIST).Methods 54 cases with GIST aged 65 years and over in Department of Gastrointestinal Surgery in Beijing Hospital from Dec.2008 to Dec.2012 were selected.31 cases underwent LECS and 23 cases underwent open surgery.Clinical data including operation time,intraoperative blood loss,complications,tumor size,length of incision,postoperative gastrointestinal function recovery time,postoperative in hospitalization and follow up data were retrospectively analyzed in patients.Results There were 17 males and 14 females in the LECS group,and the mean age was (72.8±5.9)years.29 patients underwent laparoscopic-assisted partial gastrectomy,and 2 cases underwent laparoscopicassisted endoscopic dissection in LECS group.All operations were successful with no conversion to open surgery and death.There were 13 males and 10 females in the open surgery group,and the mean age was(73.3±6.1)years.The mean operation time was longer in LECS group than in open surgery group [(120.6±32.8) min vs.(86.3±33.5) min].The mean intraoperative blood loss,mean tumor size,mean length of incision,mean time of gastrointestinal function recovery,mean postoperative hospitalization were less or smaller in LECS group than in open surgery group [(40±23.4) ml vs.(130±65.6) ml,(2.3±1.2)cm vs.(3.6 ±1.8)cm,(3.6±1.1) cm vs.(14.4±3.5) cm,(47.7± 10.4)h vs.(61.4±11.9)h,(5.1±2.3)d vs.(7.2±2.5)d,respectively,t=3.192,1.831,5.212,2.014,3.519,P=0.002,0.012,0.000,0.015,0.001].According postoperative tumor risk assessment,13 cases were at very low risk,15 cases at low risk and 3 cases at middle risk in LECS group; 4 cases were at very low risk,14 cases at low risk and 5 cases at middle risk in open surgery group.The risk degree was lower in LECS group than in open surgery group (x2 =5.63,P=0.017).During a follow-up of 5 53 months,death without GIST was found in 5 patients in LECS group and 4 cases in open surgery group.Hepatic metastasis was found in 1 case in open surgery group.Conclusions LECS is a safe and feasibility alternative approach for elderly patients with gastric GISTs.It has more minimal invasion,fast recovery and satisfaction with short term outcomes as compared with conventional open surgery.

Objective To investigate the effect of gene silencing of Y-box binding protein-1(YB-1) by RNA interference on the proliferation and migration in prostate cancer cells lines PC-3 cells .Methods YB-1 siRNAs(pGenesil-1-YB-1-1 and pGenesil-1-YB-1-2) were synthesized and transfected into cloned into the the PC-3 cells by liposome .The expressions of YB-1 were measure by RT-PCR and Western blotting .The proliferation and migration were respectively detected by MTT and Transwell method . Results ThemRNA and protein expressions of YB-1 were significantly decreased by pGenesil-1-YB-1-1 and pGenesil-1-YB-1-2 (P<0 .05) ,compared with the control group ,the inhibition ratio of mRNA expression was 36 .23% and 39 .42% respectively and the inhibition ratio of protein expression was 41 .56% and 55 .33% respectively .The proliferation and migration were significantly decreased by pGenesil-1-YB-1-1 and pGenesil-1-YB-1-2(P<0 .05) .Conclusion YB-1 gene silencing by RNA interference inhibits the proliferation and migration in prostate cancer cells lines PC-3 cells .

Objective To investigate the effect of hydrogen sulfide (H2S) on the cholesterol efflux and ATP-binding cassette transporter A1 (ABCA1) expression in foam cells .Methods RAW 264 .7 macrophages were incubated with oxidized low density lipoprotein to induce foam cells .Foam cells were incubated with H2S donor sodium hydrosulfide .Cholesterol efflux from macropha-ges was tested by labed cholesterol .The cellular levels of free cholesterol (FC) ,cholesterol ester (CE) and total cholesterol (TC) were measured by high performance liquid chromatography assays .The mRNA and protein expressions of ABCA1 were detected by Real-time PCR and Western blot .Results Compared with the foam cells ,the rates of cholesterol efflux were significantly in-creased ,the levels of TC ,FC ,CE and CE/TC ratio were significantly decreased(P<0 .05) and expression of ABCA1 was signifi-cantly increased by treatment with H2S in dose-and time-dependent manner(P<0 .05) .Conclusion H2S up-regulates of expres-sion ABCA1 and promotes cholesterol efflux in RAW 264 .7 macrophage-derived foam cells .

Objective To understand hand hygiene(HH) compliance among health care workers(HCWs) and incidence of healthcare-associated infection(HAI) in surgical patients before and after the intervention, analyze the effect of HH on cost-effectiveness of HAI.Methods From December 2012-June 2014, 78 HCWs in the department of neurosurgery of a hospital were as the intervention objects of HH compliance, 325 patients who underwent craniocerebral clean operation were as the surveyed objects, HH compliance among HCWs, incidence of HAI in surgical patients, cost of HH, and hospitalization expense before and after intervention were compared respectively.Results HH compliance among HCWs before and after intervention were 35.24% (216/613) and 73.75%(486/659)respectively (X2=180.091,P<0.001);incidence of HAI in surgical patients before and after intervention were 31.85%(50/157)and 18.45%(31/168)respectively(X2=7.782,P<0.001).Hospitalization expense before and after intervention were (89 524.90±38 856.70)￥ and (61 401.00±29 237.80)￥ respectively;average length of hospital stay before and after intervention were 36.70 days and 26.90 days respectively(t=3.296, 3.511,respectively, both P<0.01).Conclusion Improving HH compliance among HCWs can reduce the incidence of HAI and hospitalization cost, and shorten the average length of hospital stay of patients.

Objective To investigate the expression of radiation-induced IER5 protein and screen its potential interaction proteins that may participate in DNA repair process.Methods HeLa cells were irradiated with 4 Gy ionizing radiation.IER5 protein expression in whole cell lysate and in nuclear fraction were detected by Western blot at different timepoints after irradiation.3 × Flag-IER5 pCMV plasmid was constrcuted and the Flag tagged-IER5 expression was verified by Western blot.293T cells were transfected with 3 × Flag-IER5 pCMV plasmid.After irradiation the cells were collected and proteins were extracted.The IER5 interaction proteins were purified using immunoprecipitation and separated by 12％ SDS-polyacrylamide gel electrophoresis.Then the binding proteins were cut from the gel and analyzed by Mass spectrometry.Results The expression of IER5 protein began to increase 4 hour post-irradiation and its peak level was observed at 12 hour post-irradiation,and it lasted until 48 hour after irradiation.The expression level of IER5 protein in whole cell lysate and nuclear fraction were both increased.With the mass spectrometry analysis,a total of 347 proteins and 256 proteins were identified in irradiated and nonirradiated groups,respectively.Fourty one differential proteins were obtained,where 10 proteins were associated with DNA metabolic process and DNA rapair in the irradiated group and the poly(ADP-ribose) polymerase 1 (PARP1) protein was further confirmed by Western blot.Conclusions IER5 protein is an DNA damage related protein,and it may participate in DNA repair process.