Objective To investigate the effects and relevant mechanisms of exogenous Rac1 gene expression on HT1080 fibrosarcoma cell invasion across collagen barrier. Methods HT1080 fibrosarcoma cell lines that stably expressed transfected dominant negative [Rac1V12N17(HN)], constitutively active Rac1 [Rac1V12(HV)] and vector(HW) respectively were used. Structure of actin cytoskeleton was stained with Texas Red-conjugated phalloidin to show the morphological characters of the cells cultured in 3D medium containing collagen protein. Assay of cell invasion across collagen barrier was performed on a thin layer of collagen gel covered the membrane of transwell chamber, and two kinds of protease inhibitor were used to observe their effects on above-mentioned invasive assay. Gelatin substrate zymography were used to detect secreted MMP activity in the medium of cells cultured in 3D matrix. Results HT1080 cells stably expressing Racl mutant exhibited distinct morphological and invasive properties, and the increased invasive ability could be eradicated after using MMP inhibitor. Exogenous Rac1 gene expression on HT1080 fibrosarcoma cell could facilitate the activation of MMP-2 secreted in the medium of either collagen or fibrin 3D cell culture system.Conclusion The stable expression of exogenous Rac1 gene in HT1080 fibrosarcoma cells could induce aggregation of actin fiber and promote invasivc property. The enhancement of MMP-2 activation by exogenous Rac1 gene expression may be one of the relevant mechanisms.

Esophageal gastric variceal bleeding(EGVB)is a serious complication of cirrhotic portal hypertension with high mortality rate. Prevention of EGVB is an important mean to improve the survival of patients. Non-selective beta-blockers(NSBBs)is one of the first-line drugs for primary and secondary prevention of EGVB,however,only about 1 / 3 of cirrhotic patients respond to this treatment when evaluated by hepatic venous pressure gradient( HVPG). This may be related to the genetic polymorphisms of NSBBs’receptors and the metabolic enzymes. This article reviewed the progress in study on therapeutic efficacy of NSBBs and its influencing factors for preventing EGVB in cirrhotic patients.

Objective To study the of effets of hFRNK gene transfected by adenoviral vector on human colon cancer cell invasion induced by gastfin.Methods The subjects were divided into three groups:control,G17 and hFRNK group.The G17 group was treated with 100 umol/L gastrin 17 for 12 h to induce Col0320WT cells.As to hFRNK group,Colo320WT cells were infected by pAdhFRNK(MOI:100)for 48 h after transient transfection of pCR3.1-CAR for 48 h and subsequently treated with gastrinl7 for 12 h,and the control group was untreated Colo320WT cells.Expression of phosphorylated FAK(PY397)were assayed by western blot.FAK(PY397)at lamellipoda was observed with a confocal microscope.The influence of hFRNK on formation of signal complex of FAK-Src-p130Cas-Dockl80 was assayed with coimnlunoprecipitation and immunity blotting.Activity of Rac-GTPase was determined by pull down.Results Phosphorylated FAKTyr397 drastically increased with the induction of gastrin.Compared with that of G17 group,FAK (PY397)expression decreased and little FAK(PY397)was found at lamellipoda,at the same time,the signal complex of FAK-Src-p130Cas-Dockl80 did not form,and the activity of Rac decreased.Conclusion hFRNK gene may block gastrin-induced FAK phosphorylation,prevent formation of the signal complex,prevention and treatment of tumor invasion and metastasis.

Background:Cholangiocarcinoma(CCA)is a fatal digestive system tumor arising from biliary epithelium. Claudin-4,a key constituent of intercellular tight junction,is aberrantly and widely expressed in various epithelial tumors,and is correlated with tumorigenesis and tumor progression. Aims:To investigate the expression of claudin-4 in CCA and its correlation with clinicopathological characteristics of the tumor and patient’s prognosis. Methods:Immunohistochemistry was used to determine the expression rate and intensity of claudin-4 in CCA tissue and matched paracancerous tissue of 77 CCA patients. Correlation of claudin-4 expression in CCA with its clinicopathological characteristics was analyzed,and survival analysis was conducted by using Kaplan-Meier method. Results:Claudin-4 was strongly expressed in CCA tissue and mildly or weakly expressed in matched paracancerous tissue;the immunohistochemical score was significantly higher in cancerous tissue than in paracancerous tissue(9. 22 ± 3. 62 vs. 7. 12 ± 4. 26,P < 0. 05). Claudin-4 expression was significantly correlated with tumor location and tumor differentiation( P all< 0. 05 );the high expression rate was significantly higher in poorly differentiated CCA than in well or moderately differentiated ones(76. 2% vs. 50. 0% ). The 31-month accumulate survival rates of claudin-4 low-expression group and high-expression group were 29. 8% and 21. 1% , respectively( P > 0. 05). Conclusions:Claudin-4 is highly expressed in CCA and negatively correlated with tumor differentiation. It might be a novel diagnostic biomarker and therapeutic target for CCA.

Aim To explore the effect of gastrin17 on expression of ?-catenin/Tcf-4 pathway in colonic carcinoma cell line——Colo320WT.Methods Colo-320WT cells were treated with 10-8 mol?L-1 gastrin17 in time-dependent way(0、1、6、12、24、48 h),while treated with 10-6 mol?L-1 L365,260(gastrin17 receptor blocker) for 30 minutes and treated with gastrin17 again for 12 h. The expression levels of ?-catenin in TX-100 solution fraction(cytoplasmic) and TX-100 insolution fraction(cytoskeleton bound) of Colo320WT cells, and the expression levels of ?-catenin/Tcf-4 complex were detected by coimmunoprecipation and Western blot. The expression levels of c-Myc and cyclinD1 were assayed by Western blot.Results Expression levels of ?-catenin in TX-100 solution fraction were decreased apparently, but increased again by L365,260 blocking. Expression levels of ?-catenin in TX-100 insolution fraction were on the contrary. Expression levels of ?-catenin/Tcf-4 complex increased apparently.Expression levels of c-Myc and cyclinD1 in Colo320WT treated by gastrin17 were higher markedly than those of Colo320WT untreated and treated by L365,260.Conclusions Gastrin17 interacted with CCK-2R and effected significantly distribution of ?-catenin in Colo320WT and activated ?-catenin/Tcf-4 pathway which led to c-Myc and cyclinD1 high expression,the way gastrin17 decreased cell-cell cohesion and increased tumor cells invasion and metastasis further.

Objective To assess the efficacy of percutaneous transhepatie variceal embolization (PTVE) in treating esophageal and gastric variceal bleeding after esophageal-gastric devascularization with splenectomy in patients with liver cirrhosis. Methods Twenty-two patients, who had history of esophageal-gastric devascularization with splenectomy, were either underwent PTVE with TH glue (n=10) or endoscopic injection of sclerosis (EIS, n = 12) for treatment of esophageal or gastric variceal rebleeding between Nov. 2006 and Sep. 2008. The patients were followed-up for recurrent bleeding, mortality, grade of esophageal and gastric varices and liver function. Portal vein pressure was measured before and after collateral embolization in PTVE group. Results ① The patients were followed-up for 12.5 months in PTVE group and 13.4 months in EIS group. There was significant difference (P<0.05) between PTVE and EIS groups in rebleeding rate (1/10 vs 7/12) and mortality (0 vs 3/12). ② The degree of esophageal and gastric varices after embolization or EIS was improved significantly. ③ For patients with portal vein thrombosis, combination of PTVE with portal vein balloon plasty could markedly improve portal vein blood supply. ④ Neither PTVE nor EIS aggravated the liver cirrhosis. Conclusion Compared with EIS, PTVE with TH glue may be a more effiective method in the treatment of rebleeding of patients with liver cirrhosis who had accepted esophagealgastric devascularization with splenectomy.

Objective To investigate the type,clinical and imaging features of iatrogenic bile duct injury and the efficacy and safety of endoscopic and interventional radiology therapy.Methods A total of 48 patients with iatrogenic bile duct injury who have undergone endoscopic and/or interventional therapy from January 1st 2013 to June 30th 2016 were enrolled.Patients' general information,causes of injury,clinical manifestations,treatment methods,efficacy and complications were retrospectively analyzed.Results The causes of iatrogenic bile duct injury were cholecystectomy(45.8％,22/48),liver transplantation (35.4％,17/48),transjugular intrahepatic portosystemic shunt (8.3％,4/48),Roux-en-Y anastomosis (6.3％,3/48) and endoscopic retrograde cholangiopancreatography (4.2％,2/48).The most common type of iatrogenic bile duct injury was stenosis of intra/extra bile ducts (66.7％,32/48).Other types included biliary fistula(18.8％,9/48),hemobilia (10.4％,5/48) and stenosis of anastomotic stoma (4.2％,2/48).The most common clinical manifestations were jaundice (37.5％,18/48) and abdominal pain (29.2％,14/48).Other clinical manifestations were fever (14.6％,7/48),hematemesis or melena (8.3％,4/48) and abnormal drainage fluid (8.3％,4/48).Diagnosis was confirmed by angiography,cholangiography or endoscopy.The overall effective rate of minimally invasive therapy was 91.7％ (44/48) and the most common complications were fever (16.7％,8/48) and pancreatitis (10.4％,5/48).Other complications were hemobilia (2.1％,1/48),cardia dilaceration (2.1％,1/48) and biliary fistula caused by catheter shedding(2.1％,1/48).Conclusion Iatrogenic bile duct injury could occur after upper abdominal surgeries,endoscopic or interventional procedures.Early diagnosis and treatment with endoscopic or vascular interventional methods can achieve satisfying efficacy and safety.

Objectives To investigate the clinical and imaging features discriminating mass forming chronic pancreatitis (CP) from pancreatic carcinoma. Methods The clinical, radiologic, laboratory and pathologic profiles of eleven patients with mass forming CP were reviewed retrospectively. Results The patients with mass forming CP were predominantly older, male, and presented with obstructive jaundice or abdominal symptoms. Serum level of CA19-9 was slightly elevated, generally below 100 U/ml, which was lower than that of pancreatic carcinoma. The diagnostic accuracy of B-ultrasound for mass forming CP was only 18.2% and it could only be the screening method; the diagnostic accuracy of CT was 45.5%, and the main CT features included dilation of main pancreatic duct and the branches, as well as pancreatic pseudocysts or calcification. The findings of MRCP and EUS were irregular dilation of main pancreatic duct and common bile duct, which were similar to pancreatic carcinoma. ERCP could provide imaging characteristics of biliary and pancreatic duct, and the nature of obstruction, the stricture and dilation of biliary and pancreatic duct. When combined with CT scan, ERCP was useful for differentiation from pancreatic head cancer. According to histopathology, a few mass forming CP was autoimmune pancreatitis. Conclusions Mass forming CP is associated with clinical and radiologic features that are similar to those of pancreatic carcinoma. It is necessary to carefully evaluating clinical and imaging findings, laboratory findings and histopathological examinations.

Objective To investigate the correlation between liver and spleen stiffness measured by acoustic radiation force impulse (ARFI) and hepatic venous pressure gradient (HVPG),and to evaluate its efficiency in the diagnosis of portal hypertension.Methods From April 2014 to March 2016,20 cases underwent HVPG measurement because of liver cirrhosis were enrolled.Before HVPG measurement,liver and spleen stiffness were assessed with ARFI.The correlation between HVPG and age,alanine aminotransferase (ALT),aspartate aminotransferase (AST),total hilirubin,serum albumin,platelet count,prothrombin time,aspartate aminotransferase to platelet ratio index (APRI) score,Child-Pugh score,model for end-stage liver disease (MELD) score,liver stiffness and spleen stiffness were analyzed.Pearson correlation and Spearman rank correlation were performed for statistical analysis.Results HVPG,liver and spleen stiffness were successfully measured in all 20 patients.The mean liver stiffness was (1.78±0.29) m/s,the mean spleen stiffness was (3.37±0.44) m/s and HVPG was (16.10±5.14) mmHg (1 mmHg=0.133 kPa).Age,ALT,AST,total bilirubin,serum albumin,platelet count,prothrombin time,APRI score,Child-Pugh score and MELD score were all not correlated with HVPG (all P＞0.05).But HVPG was positively correlated with liver and spleen stiffness (r=0.449,P=0.047;r=0.487,P=0.030).In the diagnosis of HVPG≥12 mmHg,the area under curve (AUC) of liver stiffness was 0.875,the optimal cut-off value was 1.77 m/s,the sensitivity was 68.6 ％ and the specificity was 100.0％.In the diagnosis of HPVG≥20 mmHg,the AUC of liver stiffness was 0.798,the optimal cut off value was 1.85 m/s,the sensitivity was 100.0％ and the specificity was 68.8％.The AUC of spleen stiffness was 0.820,the optimal cut-off value was 3.23 m/s,the sensitivity was 100.0 ％ and the specificity was 56.3％.Conclusion In patients with liver cirrhosis,liver stiffness and spleen stiffness assessed by ARFI are positively correlated with HVPG and therefore ARFI has certain application value in the noninvasive diagnosis of portal hypertension.

Objective To explore the effects of different treatments on prognosis of patients with Gynura segetum caused hepatic vein occlusion disease (HVOD).Methods From July 2008 to January 2016,85 patients with Gynura segetum caused HVOD were enrolled and received treatment of liver function protection and microcirculation improvement.According to different treatment options,patients were divided into non-anticoagulation group,nowanticoagulation transfered to transjugular intrahepatic portosystem stent-shunt (TIPS) group,anticoagulation group,anticoagulation transfered to TIPS group and anticoagulation-TIPS step-by-step treatment group.The efficacy of each group was observed.Chi square test was performed for statistical analysis.Results Among 22 patients who received nonanticoagulation treatment,six (27.3％) patients were cured and 14 (63.6％) patients died during the treatment period;besides two (9.1 ％) patients received TIPS because of ineffective treatment and achieved longterm survival.Among 63 patients treated with combination of low-molecular-weight heparin and warfarin,six (9.5％) patients died and 36(57.1％) patients were cured.The cure rate was higher than that of nonanticoagulation group (x2 =5.820,P=0.016).Other 21 patients received TIPS treatment,achieved longterm survival except one patient died from surgical complications.The cure rate of anticoagulation-TIPS step treatment group was 88.9 ％ (56/63) which was higher than that of non-anticoagulation group,and the difference was statistically significant (x2 =31.350,P＜0.01).Conclusions Compared to conventional liver function protection treatment and symptomatic treatment,anticoagulation therapy significantly increases the cure rate of patients with Gynura segetum caused HVOD.Anticoagulation-TIPS step-by-step treatment further improves the cure rate.