Objective To design and construct the vector of protein-rich tyrosine kinase 2 small RNA interference and investigate the effect of the recombinant plasmid on the proliferation of adult rat cardiac fibroblasts. Methods The pAVU6+27-PYK_2 siRNA expression vector was constructed by gene recombination, then transfected into the cultured adult rat cardiac fibroblasts by DOTAP method. PYK_2 mRNA and protein in cardiac fibroblasts were determined by RT-PCR and Western blotting respectively. Cell proliferation was tested by MTT and ~3H-TdR incorporation. Results The pAVU6+27-PYK_2 siRNA expression vector was successfully constructed. The recombinant plasmid can inhibit the proliferation of adult rat cardiac fibroblasts, as compared with control. Conclusion pAVU6+27-PYK_2 siRNA expression vector is efficient to suppress DNA synthesis of cardiac fibroblasts, which will be beneficial to further study of myocardial fibrosis.

Objective To assess the efficacy of the laryngeal mask airway i-gel (LMA i-gel) in patients umdergoing laparoscopic cholecystectomy. Methods One hundred and twenty ASA Ⅰ or Ⅱ patients of both sexes,aged 34-62 yr, weighing 45-90 kg, undergoing elective cholecystectomy using fiberoptic laparoscope, were randomized into 2 groups (n =60 each): LMA i-gel group (group I) and LMA ProSeal group (group P). After induction of anesthesia with sufentanil 0.2 μg/kg, propofol 2.5 mg/kg and vecuronium 0.1 mg/kg, LMA i-gel and LMA ProSeal were inserted in group I and P respectively. A gastric tube was inserted through the drain tube of the LMAs. The number of attempts, success rate, success rate of gastric tube placement, airway seal pressure, SpO2 ,PETCO2, peak airway pressure and complications (hypoxemia, nausea and vomiting, choking hoarseness, sore throat and regurgitation of gastric contents) were recorded. The fiberoptic laryngoscope scores were assessed after sccessful LMA placement. The anesthesia time, duration of surgery, extubation time and emergence time were also recorded. Results There was no significant difference in the anesthesia time, duration of surgery, extubation time and emergence time between the two groups ( P ＞ 0.05). The SpO2, PETCO2 and peak airway pressure were within the normal range during operation in both groups. The first attempt success rate of LMA placement and fiberoptic bronchoscopic scores were significantly higher, and the LMA placement time was significantly shorter in group Ⅰ than in group P (P ＜ 0.05). The success rate of LMA and gastric tube placement was 100% in both groups. The incidence of sore throat was significantly lower in group I than in group P ( P ＜ 0.05). Conclusion LMA i-gel can provide adequate ventilation during operation with less complications and can be used effectively for cholecystic laparoscopic surgery.

Between January 2011 and December 2011,86 patients undergoing operation for anorectal disease were randomized in a double-blind manner to receive a mixed injection of methylene blue,ropivacaine and sodium chloride (observation group,n =43) or traditional analgesic methods (control group,n =43).Patients in observation group had less visual analog scale (VAS) scores of pain at 6-48 h postoperation than control group (P ＜0.05).The scores had no significant difference at 72 h postoperation between two groups (P ＞ 0.05).The total postoperative complications (8,19％) and hospitalization duration [(5.3 ± 2.2) days] in observation group were significantly less than those [15,35 ％ and (6.4 ±1.7) days] in control group (P ＜ 0.05).A mixed injection of methylene blue and ropivacaine intraoperatively can relieve the postoperative pain of patients with anorectal disease and reduce the incidence of postoperative complications and the duration of hospitalization.

Objective To study the effects of postoperative immune-enhancing enteral nutrition (IEEN)on the liver function and postoperative infectious complications after hepatectomy in patients with hepatocellular carcinoma.Methods Between January 2010 and December 2011,96 patients undergoing hepatectomy for hepatocellular carcinoma were randomized in a double-blind manner to receive postoperative enteral nutrition with IEEN (IEEN group,n =48) or standard enteral nutrition (control group,n =48) for 5 days,respectively.Calorie and nitrogen of the same quantity were provided for both groups from the first day after surgery.The liver function tests and routine blood tests were performed.The postoperative hospital stay and postoperative infectious complications were also compared between these two groups.Results The serum albumin,prealbumin,alanine aminotransferase,and total bilirubin levels had no significant differences between the two groups (all P ＞ 0.05).On the 6th day after operation,the level of total lymphocyte count was significantly higher in the IEEN group than that in the control group [(2106 ± 1057) vs.(1648 ± 1079) / μL,P =0.038).The postoperative hospital stay was (10.3 ±5.6) days in the IEEN group and (13.4 ±6.7) days in the control group (P =0.016).Postoperative infectious complications were observed in 8 (16.7％) patients in the IEEN group and 17 (35.4％) patients in the control group (P =0.036).Conclusion Early postoperative IEEN can improve the clinical outcomes of liver cancer patients after hepatectomy.

Objective To study the role of survivin in the incidence of pulmonary hypertension secondary to congenital heart disease (CHD) of left-to-right shunt. Methods Forty-nine cases were divided into 4 groups according to mean pulmonary artery pressure (mPAP), that is, non-hypertension group, mild hypertension group, medium hypertension group, and severe hypertension group. Their lung tissue samples were collected during heart surgery. TUNEL technique and immunohistochemical analysis were respectively used to detect nuclear DNA fragment of apoptotic cells and the expression of survivin, bcl-2 and bax. Western blotting also was conducted to detect survivin expression. Results The pulmonary wall of CHD patients with medium or severe pulmonary hypertension became significant incrassate. The apoptotic index and expression level of bax were significantly decreased but surviving and bcl-2 expressions were significantly increased in CHD with medium or severe pulmonary hypertension. Survivin expression was significant correlation with percentage media wall thickness and apoptotic index. Conclusion Apoptosis associated with survivin plays an important role in pulmonary artery remodeling after pulmonary hypertension of CHD.

Objective To approach the expressions and the roles of connective tissue growth factor (CTGF) and membrane type 1 matrix metalloproteinases (MT1-MMP) in valve disease with pressure overload which induces extracellular matrix remodeling of left ventricle. MethodsOf 32 patients, 16 cases were pressure overload group (PO), who had the multiple valve disease with predominately aortic valve stenosis, having a ring diameter of aorta valve less than 1.3 cm, cross valve pressure gradient equal or more than 40 mmHg, and valvular regurgitation less than 4.0 cm2; The other 16 cases, as mitral stenosis group (MS), were simple mitral stenosis patients with single valve replacement. Meanwhile, 5 normal individuals served as control, who died from accident. Echocardiography was used to analyze the left ventricular function and detect the hypertrophic level of the left ventricle. Left ventricle muscle samples were obtained during operation. Histological features were studied by Masson staining, and collagenous contents were quantitated with a computer-assisted imaging analysis system. The mRNA expressions of CTGF and MT1-MMP were detected with RT-PCR. ResultsConcentric hypertrophy was observed significant in PO group, but myocardial hypertrophy was not found in MS group. Compare to the MS group and control, PO group had significantly more collagenous contents in left ventricle, thickened vessel wall, and narrow lumen of blood vessel (P0.05), but CTGF mRNA expression was increased in MS group compare to control (P

Objective To explore the causation,diagnosis and management of iatrogenic bile duct injury(BDI) of laparoscopic cholecystectomy(LC).Methods A total of 1 656 patients undergoing laparoscopic cholecystectomy in our department during the last 7 years were included in this study for retrospective analysis.Results There were 274 patients with gallbladder polyps,168 patients with acute calculous cholecystitis and 1214 patients with chronic calculous cholecystitis.There were 15 BDIs associated with LC(0.91%).A total of 8 BDI patients were diagnosed during cholecystectomy.The remaining 7 BDI patients were diagnosed postoperatively.The intraoperative diagnosis of BDI was made on the discovery of bile leakage or double biliary stump during cholecystectomy.Clinical features,diagnostic abdominocentesis and imaging findings formed the basis of diagnosis of BDI postoperatively.One BDI patient was treated by repairing the injuried common bile duct with a T-tube drinage.Four BDI patients were treated by end-to-end anastomosis of injuried bile duct,and one of the four patients was reoperated with Roux-en-Y hepaticojejunostomy because of bile leakage.The remaining 10 BDI patients were treated by Roux-en-Y hepaticojejunostomy,and good results were achieved in all of these patients.Conclusions There is no relationship between the etiology of gallbladder disease and BDI during laparoscopic cholecystectomy.Good results can be achieved if BDI is diagnosed early and treated properly during or after operation.Roux-en-Y hepaticojejunostomy is the primary operation method for treating BDI.

Objetive To investigate the effect of Fasudil on the hepatic ischemia repeffusion injury on rats of obstructive jaundice.Methods 160 mature SD rats were randomly assigned into 2 large groups,namely experimental group (Group A) and control group (Group B).While each large group was subdivided into 4 subgroups,including sham operation subgroups(Group A1 and B1),obstructive jaundice model subgroups(Group A2 and B2),ischemia reperfusion model subgroups (Group A3 and B3)and obstructive jaundice plus ischemia reperfusion model subgroups (Group A4 and B4).In Group A2 and B2,obstructive jaundice models were made on rats by double ligating and cutting the common bile ducts.In Group A3 and B3,the ischemia reperfusion injury models were built by blocking the hepatic portal vessel for 30 minutes before reperfusion.In Group A4 and B4,the ischemia reperfusion injury models were built in 1 week after building the obstructive jaundice models.For Group A,10 mg/kg fasudil was intraperitoneally injected at 30 minutes before ischemia.For Group B,equal amount of physiological saline was injected.Samples were gained on zero hour,1 hour,2 hour and 6 hour post reperfusion.The serum indexes for liver function and serum Endothelin 1 level were tested and analyzed through SPSS.Pathological changes of liver were viewed correspondingly.Results Compared to control groups (Group Bs),experimental groups (Groups As) have better liver function,lower Endothelin 1 level (P ＜ 0.05),as well as less severe microscopic histological damage.Conclusion Fasudil can effectively protect rats of obstructive jaundice from hepatic ischemia reperfusion injury.

Objective　To explore the injurious effect of the serum drawn from patient under cardiopulmonary bypass (CPB) on pulmonary surfacant-associated protein A(SP-A) and the preventive effect of pentoxifylline (PTX) on the protein. Methods　The cultured rat alveolar epithelial cell type Ⅱ(AT-Ⅱ) were incubated with CPB serum to observe the change of the cell morphology, and the expressions of SP-A and SP-A mRNA. Results　Traumatic changes and the decrease of SP-A and SP-A mRNA expressions were found in AT-Ⅱ cells cultured with CBP serum. PTX exerted protective effect on the cells. Conclusion　The serum after CPB can directly injure rat AT-Ⅱ cells in vitro, and inhibit the SP-A expression at transcription and translation levels, which probably is an important reason for the quantitative and qualitative abnormality of pulmonary surfactant after operation. PTX may alleviate the inhibitory effect of CPB serum on SP-A.

Objective To investigate the effects of isoliquiritigenin on the invasive ability of human gastric carcinoma SGC7901 cells, and its molecular mechanisms thereof. Methods The logarithmic phase human gastric carcinoma SGC7901 cells were divided into control group (normal cell culture fluid) and isoliquiritigenin group (isoliquiritigenin solu?ble in cell culture fluid, the concentrations were 10, 25, 50 and 100 μmol/L respectively). Each group had four repeated holes. The proliferation of SGC7901 cells were detected with MTT assay after 24 h, 48 h and 72 h of culture. The experimen?tal drug concentration and action time were researched for the subsequent experiments. The in vitro invasion abilities of SGC7901 cells were assessed with Transwell test. The expression levels of MMP9, Akt and P-Akt were detected by Western blot assay. Results The proliferation of SGC7901 cells were inhibited by 10μmol/L isoliquiritigenin, which can be signifi?cantly inhibited by 25, 50 and 100μmol/L isoliquiritigenin in a concentration-dependent and time-dependent manner. The half inhibitory concentrations (IC50) of 24, 48 and 72 h were 52.48, 44.49 and 32.50μmol/L, respectively. Therefore, the 25, 50 and 100μmol/L isoliquiritigenin were selected as the subsequent experimental drug concentration, and 24 h was used as the action time. Compared with the control group (209.75±9.29), the membrane cell number of 25μmol/L (138.50±10.15), 50μmol/L (89.50 ± 16.56) and 100μmol/L (45.00 ± 8.08) decreased gradually (F=267.948,P<0.05). There was no signifi?cant difference in the expression level of Akt protein between four groups (F=1.492). The expression levels of P-Akt and MMP9 were gradually decreased with the increase of the isoliquirigenin concentration (F=359.219 and 431.324,P<0.05). Conclusion Isoliquiritigenin can obviously inhibit invasion ability of SGC7901 cells, which may be related to the down reg?ulation of the signal transduction pathway protein PI3K/Akt and the down steam protein MMP9.