Objective To observe the effect of SIRT1 on the proliferation of ovarian cancer CAOV3 cells and its possible mechanism. Methods Ovarian cancer CAOV3 cells were transfected the Lipofectamine 2000 and were divided into normal control group,negative control group and SIRT1-siRNA group.The mRNA and protein expression levels of SIRT1 were detected by RT-PCR and Western blot.MTT method was used to detect the proliferation of RNA interference at 24h,48h and 72h,respectively.The distribution of CAOV3 cell cycle phase after RNA interference of SIRT1 for 72 hours was detected by flow cytometry.SIRT1,p53,p21 protein expression and p53 acetylation status were detected by Western blot with RNA interference of SIRT1 after 72 hours.The interaction between SIRT1 and p53 protein was verified by immunoprecipitation in CAOV3 cells.Results Compared with negative control group,the expression of mRNA and protein of SIRT1 in SIRT1-siRNA group decreased after transfection of SIRT1 for 72 hours.The survival rate of CAOV3 cells in SIRT1-siRNA group decreased significantly at 48 hours and 72 hours (P<0.01).The proportion of Sphase cells in CAOV3 cells transfected with SIRT1-siRNA for 72h was significantly decreased (P<0.01),while the G1 phase cells were significantly increased (P<0.01),and the cells arrested in the G1 phase.The expression of p53 and p21 proteins were significantly increased and the p53 was in higher acetylation status after transfection with SIRT1-siRNA for 72 hours ( P<0.01 ) .SIRT1 was interacted with p53 protein in CAOV3 cells confirmed by co-immunoprecipitation. Conclusion Downregulation of SIRT1 expression can inhibit the proliferation of ovarian cancer CAOV3 cells, which maybe connected with p53 at high acetylation status,and enhancing p53-dependent p21 transcriptional activation,thus blocking the cell cycle and inhibiting the cell proliferation.

Objective To discuss the implication of ectopic opening of the pancreaticobiliary duct in the performance of cholangiopancreatography (ERCP) and the diagnosis and treatment of cholangio-pancreatic disease. Methods We summarize the data of 1000 cases of ERCP in the past four years. Five patients were found with ectopic opening of the pancreaticobiliary duct. The positions, characteristics of ectopic opening and the characteristics of the mucosa disease around the ectopic papilla and cholangio-pancreatic disease were analyzed. Results In the 5 patients with ectopic opening of the papilla, one had the opening in the pyloric channal, two in the duodenal bulb, the other two in the distance of the descending part of duodenum. Mucosa erosion was found in the patients with ectopic opening in the pyloric channal and in the duodenal bulb. Conclusion The ectopic opening of the pancreaticobiliary duct may affect the finding of the papilla, but may not affect the performance of ERCP. The incidence of the cholangiopancreatic disease and the mucosa disease around the ectopic papilla will increase in the patients with ectopic opening.

Objective To explore effective methods of nursing quality evaluation. Methods Guided by human-centered thinking,the new DRG quality indicators were formulated based on clinical investigation,literature review and expert consulta-tion. Then,the standards of the new indicators for radical masteetomy were identified by Delphi method and the reliability and validity were tested in 30 patients with radical mastectomy. Results The two new quality indicators were illness observation and health education. The Cronbach'α was 0.82 and 0.88 respectively. The results of expert consultation showed high content validity. Moreover,the new quality indicators got high commend by involved nurses. Conclusion The new quality indicators will lead nursing work to illness observation and health education,which will more benefit to reflect the true work of nurses and improve the internal quality of nursing.

Objective To apply failure mode and effect analysis (FMEA) to reduce nursing risk of peripherally inserted central catheter (PICC).Methods 100 patients with PICC in central hospital of Minxing District of Shanghai were named as the experimental group,failure mode and effect analysis were performed in this group to obtain the value of risk priority number (PRN) and array them according to the RPN value to work out improved plan systematically and put it into practice to guide the clinical operation and nursing care.98 PICC patients in Xinzhuang health service center and Tumor Hospital of Minhang district were set as the control group,which were subjected to traditional nursing.All statistical analysis were done by SPSS 11.0 software package,the differences of the incidence of PICC-related nursing risk between the two groups due to the implementation of FMEA was analyzed by X2 test.Results Application of FMEA could reduce the nursing risk of PICC,After the application of FMEA,the incidence of PICC-related complications such as focal or systemic infection,puncture site oozing and bleeding,skin allergies,mechanical phlebitis,too deeply placed catheter,catheterization into cervical veins and reflexed into axillary vein,catheter occlusion,catheter migration,difficult removal of catheter,puncture failure in the experimental group decreased obviously,the differences had statistical meaning.Conclusions The application of failure mode and effects analysis can reduce the nursing risk of PICC,decrease medical disputes,alleviate the suffering of patients,reduce medical costs caused by PICC-related complications,and improve the satisfaction degree of patients and the quality of nursing care.

Objective To investigate the influence of endoscopic retrograde cholangiography (ERC), endoscopic retrograde pancreatography (ERP), endoscopic retrograde cholangiopancreatography (ERCP), endoscopic sphincterotomy (EST) and endoscopic biliary stenting on postoperative pancreatitis. Methods 412 patients referred to ERCP were divide into 7 groups, there were both biliary and pancreatic ducts group (ERCP), biliary duct contrast filling group (ERC), pancreatic duct contrast filling group (ERP), ERCP plus biliary stenting group (stent), ERC plus stent, ERCP plus EST and stone extraction (SE) group, and ERC plus EST and SE group. And the differences of postoperative serum amylase in 4 hours and in 24 hours as well as clinical symptoms were compared among different groups. Results The incidence of postoperative hyperamylasaemia in 4 hours and 24 hours were 17.7% and 4.4% respectively. The incidence of postoperative acute pancreatitis was 3.9%, and ERP group had the highest incidence of postoperative acute pancreatitis among the 7 groups. Conclusions Repeated pancreatic duct contrast filling during ERCP manipulation is the main risk factor for postoperative pancreatitis, and therapeutic ERCP such as EST, stent and SE does not increase the incidence of postoperative pancreatitis.

Objective To investigate the localization and mRNA expression of dopamine receptor subtype D_4 and D_5 mRNA in human stomach and duodenum by in situ hybridization. Methods The oligonucleotide probe was synthesized and labeled with digoxin, and then hybridized with human gastric and duodenal samples, which were stored in liquid nitrogen, fixed in hypothermia, and embedded in paraffin. The results were recorded and quantitatively analyzed using the pathological image computer analysis system. Results Dopamine receptor subtype D_4 and D_5 mRNA both expressed in human stomach as well as duodenum. They distributed mainly in the interstitial of mucosal glands and the lamina propria near lamina muscularis mucosa. The mRNA expression value of D_4 was higher than that of D_5 in both stomach and duodenum. In stomach, the number of positive target cells was 41.29?5.06 vs. 26.25?5.82 ( P

AIM:To evaluate the effect of bFGF on the apoptosis and the expression of glucose regulated protein 78(GRP78) in ovarian cancer CAOV3 cells.METHODS:Starvation induced cell apoptosis was conducted.After treatment with bFGF,the cell cycle and apoptosis were determined by FACS analysis and Annexin V/PI staining,respectively.The expression of protein kinase B(PKB) and GRP78 were detected by Western blotting and RT-PCR.RESULTS:Compared to starvation group,the cells treated with bFGF were still viable and increased activation of PKB and high expression of GRP78 were observed,which were prevented by PKB inhibitor wortmannin effectively(P

Objective To investigate the safety of endoscopic suhmucosal dissection (ESD) in elderly patients with lesions of gastroesophageal mucosa. Methods Thirty-six elderly patients with flat elevation lesion of gastroesophageal mucosa were studied from July 2007 to May 2008. There were 14 cases with dysplasia, 9 cases with early carcinoma and 13 cases with leiomyoma confirmed by pathologic biopsy and endoscopic ultrasonography. All the patients were treated under consciousness condition, and the focus of lesion were completely resected with improved hook knife, IT knife and electrocoagulation or electro-cutting technique. The wound surface was managed with noradrenaline added in ice physiological saline, argon beam coagulator and metal clips. After operation, they were treated by abrosia, acid suppression and mucosa protection. The conditions of wound healing and residual lesions were reexamined four weeks after operation. Results Among the 36 patients (19 males and 17 females), there were 29 patients(80.6%) with complete focus ablation by ESD and 7 patients (19.4%) with resection by snare. Three cases (8.3%) of perforation occurred during operation and the perforations were treated with metal clips. After inserting stomach tube for fasting and anti-infection treatment for 5-7 days, all the three cases were recovered. One case(2.8%)had hematemesis 2 hours after operation and bleeding was stopped by endoscopic metal clip. Conclusions ESD is a safe and effective therapeutic tool in elderly patients with lesions of gastroesophageal mucosa.

Objective Corneal endothelial decompensation is caused by many corneal diseases. It often results in severe clinical complications. Endothelial keratoplasty (EK) is a new therapy for corneal endothelial decompensation. This study aimed to investigate a new approach to establishing corneal endothelial decompensation animal model with Descemetorhexis technique in order to better understand the tissue response to EK. Methods Thirty New Zealand white rabbits were randomly divided into three groups according to different surgical procedures; corneal endothelial cells (CEC), Descemet's membrane and corneal endothelial cells (DM + CEC) as well as Descemet' s stripping with endothelial keratoplasty(DSEK) group and 10 eyes for each. The right eyes of rabbits were as surgery eyes. Other 10 rabbits were as DSEK donors. Corneal transparency, anterior chamber response and graft location were examined once per day for two weeks under the slit lamp. Comeal thickness was measured by ultrasound biomicroscope. Corneal endothelial cells were analyzed using vital staining with alizarin red and trypan blue in 2, 4 and 8 weeks after operation. Results The cornea in DM + CEC group remained opaque throughout the observation period. In CEC and DSEK group, corneal clarity was gradually restored and corneal thickness was significantly less than that in the DM + CEC group during the postoperative 8 weeks. There were significant differences in corneal thickness between the DM + CEC group and CEC group or DSEK group during the postoperative 8 weeks (P ＜0. 05). The vital staining showed that most Descemetorhexis area was not covered by endothelial cells even 2 months after surgery. Conclusion A new corneal endothelial decompensation model is successfully established for the study of corneal endothelial keratoplasty, which is helpful for understanding the wound-healing of rabbit corneal endothelium after Descemel' s membrane damage.

To observe the chelation of GRP78 with lead (Pb) and its localization changes, astroglial cells from Wistar rat brain were primarily cultured in medium with acetate Pb. The processes were terminated at different time points. The immunoprecipitation (IP) and Western blotting were used for GRP78 purification and expression and the Pb concentration was determined by employing atomic absorption spectrophotometry (AAS). The localization change of GRP78 was observed with colloid gold immunoelectron microscopy. The results showed that the expression of GRP78 was increased significantly in the cells treated with 1.0 mumol/L acetate Pb for 24 h and peaked at 96-192 h (P<0.01), and at the 12th day, the expression of GRP78 began to decrease but was still higher than normal (P<0.05). Pb content started to increase when cells were treated by acetate Pb for 24 h, and the peak appeared at 8 day (P<0.01), and then Pb content decreased gradually, but was still higher than normal (P<0.05). GRP78 protein expression began to remarkably increase when it transferred from ER to the cytosol around the nuclei 24 h after treatment with Pb. It is concluded that GRP78 in astroglia could strongly chelate with Pb ions and it might be a target protein of Pb.