Objective To explore the expression of cullin 4A (CUL4A) in cholangiocarcinoma tissues and its clinical significance.Methods Primary fresh cholangiocarcinoma tissues (n =35) and normal bile duct tissues (n =15) from patients who underwent curative surgery in Binzhou People's Hospital of Shandong Province from February 2011 to December 2013 were collected.The expressions of CUL4A mRNA were detected by quantitative real-time PCR (qRT-PCR).Then,cholangiocarcinoma tissues (n =72) and normal bile duct tissues (n =36) from patients who underwent curative surgery in Binzhou People's Hospital of Shandong Province from January 2008 to January 2014 were collected.The expressions of CUL4A protein were tested by immunohistochemistry.The relationships between the expression of CUL4A and the patients' clinicopathologic features and prognosis were analyzed.Results The expression of CUL4A mRNA in cholangiocarcinoma tissues was obviously higher than that in normal bile duct tissues (3.876 ±0.975 vs.1.216 ±0.265),and the difference was statistically significant (t =12.23,P ＜ 0.001).The positive rates of CUL4A protein in cholangiocarcinoma and normal bile duct tissues were 70.8％ (51/72) and 2.8％ (1/36) respectively,and the difference was statistically significant (x2 =44.524,P ＜ 0.001).The expression of CUL4A protein in cholangiocarcinoma was related to the differentiated degree (x2 =4.341,P =0.037),neural invasion (x2 =8.326,P =0.004),TNM stage (x2 =7.745,P =0.005),lymph node metastasis (x2 =3.869,P =0.049) and vascular invasion (x2 =5.555,P =0.018).Survival analysis results showed that the median survival time of CUL4A positive patients was significantly shorter than that of CUL4A negative patients (32.40 months vs.51.30 months),and the difference was statistically significant (x2 =6.561,P =0.011).Conclusion The expression of CUL4A in cholangiocarcinoma tissues is higher than that in normal bile tissues.CUL4A plays an important role in the process of tumor invasion and metastasis and indicates poor prognosis,which may become a potential target for the diagnosis and therapy of the cholangiocarcinoma.

Objective To investigate the expressions of cancerous inhibitor of protein phosphatase2A (CIP2A),phosphatidylinositol 3-kinase(PI3K),and survivin in pancreatic carcinomas and their clinical significance.Methods The expressions of CIP2A,PI3K,and survivin proteins were tested by immunohistochemistry in 64 cases of pancreatic carcinomas and adjacent paracancerous tissues.Results The positive rate of CIP2A in pancreatic carcinomas was significantly higher than adjacent paracancerous tissues (70.3％ vs 5.6％,P ＜0.05).Significant difference was observed in the expression rate of PI3K between the patients with pancreatic carcinomas and paracancerous tissues (73.4％ vs 8.3％,P ＜0.05).Significant difference was also observed in the expression rate of survivin between the patients with pancreatic carcinomas and paracancerous tissues (75.0％ vs 2.8％,P ＜0.05).CIP2A,PI3K,and survivin were significantly differentially expressed in pancreatic carcinoma among different tumor differentiation,tumor node metastasis (TNM) stage,and neural invasion and lymph node metastasis (all P ＜ 0.05).Spearman correlation analysis showed significantly positive correlation between the expressions of CIP2A and the others (PI3K and survivin) (both P ＜0.05),and between the expressions of PI3K and surviving (P ＜0.05).Conclusions CIP2A was involved in the development of pancreatic carcinomas and might activate the PI3K/Akt/survivin pathway.Our data identified CIP2A as a critical oncoprotein involved in cell proliferation,invasion,and metastasis.It could serve as a therapeutic target for pancreatic carcinomas.

Objective To investigate the diagnosis and treatment of ectopic opening of the common bile duct in the duodenal bulb.Methods The clinical data of 3 patients who were admitted to the Binzhou People's Hospital and 9 patients who were admitted to the Tianjin People's Hospital from January 2006 to December 2013 with ectopic opening of the common bile duct in the duodenal bulb were retrospectively analyzed.Seven patients had choledocholithiasis and 5 had stenosis at the end of common bile duct.The medical histories and clinical features in patients were analyzed and routine blood test and serum liver function test were done.All the patients received the endoscopic retrograde cholangiopancreatography (ERCP) examination and were cured.All the patients were followed up via outpatient examination and telephone interview up to August 2014.Results Six patients had histories of cholangitis recurrence and 2 had histories of duodenal ulcer recurrence.All the patients had pain in the right hypochondriac region of the abdomen.Seven patients had fever,chills,skin yellowing sclera and tenderness in the right hypochondriac region of the abdomen.The levels of alkaline phosphatase (ALP) and glutamyltranspeptidase (GGT) in 11 patients,the levels of TBil and DBil in 8 patients and the count of WBC in 7 patients were increased.(1) The results of ERCP showed as follows:there was no papillar opening at the second and third segment of duodenum.The crack-like opening located at the duodenal post-bulb with rough and erosive mucosal surfaces and intermittent outflow of bile.Duodenal ulcer was detected in 5 patients and duodenal bulb metamorphosis in 3 patients.All the 12 patients received successfully intubations.(2)The results of retrograde cholangiography showed as follows:the end of common bile duct of 12 patients was taper and sickle-shaped.Intra-and extrahepatic bile duct dilation was detected in 10 patients,choledocholithiasis in 7 patients and clear findings for the pancreatic duct in 5 patients.Among the 12 patients,8 received balloon dilation (5 with stenosis at the end of common bile duct,3 with choledocholithiasis),3 received laparoscopic common bile duct exploration (LCBDE) combined with cholangioenterostomy due to diameter of stone more than 1.5 cm and ectopic opening stenosis of the common bile duct in the duodenal bulb.One patient was treated by percutaneous transhepatic cholangiography (PTC) lithotomy of common bile duct after unsuccessful ERCP without bleeding and pancreatitis-related complications.The symptoms of cholangitis in 3 patients were alleviated after balloon dilation,2 patients had recurrence of cholangitis and were cured by Roux-en-Y cholangioenterostomy.The mean open surgery time and mean duration of postoperative hospital stay in 5 patients were 85 minutes (range,60-150 minutes) and 10 days (range,8-14 days),respectively.All the 12 patients were followed up with a median time of 38 months (range,8-90 months).During the follow-up,10 patients survived well without recurrence of cholangitis and cholelithiasis.Two patients had recurrence of cholangitis at postoperative month 2 and month 14,including 1 patient with the recurrence of common bile duct sand-like stones,and they were readmitted to hospital and treated by Roux-en-Y cholangioenterostomy without recurrence by follow-up.Conclusions The clinical symptoms of ectopic opening of the common bile duct in the duodenal bulb included recurrence of cholangitis,duodenal ulcer history,pain in the right hypochondriac region of the abdomen,skin yellowing sclera,abnormal liver function,crack-like openings in the duodenal bulb by ERCP examination with outflow of bile,cholangiography-guided taper and sickle-shaped end of common bile duct.The treatment should be aimed at the concomitant diseases.

Factor VII Deficiency ; complications ; Humans ; Liver Cirrhosis ; complications ; Male ; Middle Aged

Objective To investigate the relationship of Twist induced epithelial mesenchymal transitions (EMT) during pancreatic carcinoma progression.Methods The expressions of Twist,N-cadherin and Vimentin proteins were tested by immunohistochemistry in 42 cases of pancreatic carcinomas and 26 cases of adjacent paracancerous tissues.Results The positive rates of Twist,N-cadherin and Vimentin in 42 cases of pancreatic carcinomas were 76.2％,59.5％ and 57.1％,respectively.The positive rate of Twist,N-cadherin and Vimentin were all significant between pancreatic carcinomas and paired tumor-adjacent paracancerous tissues (all P ＜0.001).The differences of the expression of Twist,N-cadherin and Vimentin in pancreatic carcinoma of tumor differentiation,TNM stage,neural invasion and lymph node metastasis were significant (all P ＜ 0.05).Significantly positive correlation was found between the expression of Twist and the others (N-cadherin and Vimentin) by using spearman correlation analysis (r =0.506,P =0.001 ; r =0.417,P =0.006).Significantly positive correlation was found between the expression of N-cadherin and Vimentin by using spearman correlation analysis(r =0.462,P =0.002).Conclusions Twist signaling pathway activation might mediate pancreatic epithelial cells to EMT and then promote pancreatic carcinoma invasion and metastasis.

Objective To investigate the effects of MMI-166 on apoptosis and apoptosis-related protein expression of human pancreatic cancer SW1990 cells and its transplanted tumor,and explore possible mechanism.Methods The human pancreatic cancer xenograft model was constructed by using human pancreatic cancer SW1990 cells.Tumor-bearing nude mice were randomly divided into control and MMI-166 groups,and they were treated with normal saline or MMI-166 (200 mg · kg-1 · d-1) for 28 days.Apoptosis index (AI),p53,c-Myc,Bax,Bcl-2,Survivin,Caspase-1,Fas proteins were detected by deoxynucl-eotidyl transferase-mediated nick end labeling (TUNEL method) and Western blot.MMI-166 of different concentrations (0,50,100 μg/ml) were used to treat human pancreatic cancer SW1990 cell for 24 h.The c-Myc,Survivin proteins expressions were measured by Western blotting.Results Apoptosis index in MMI-166 group was 81.1 ±7.9,which was significantly higher than that in control group (21.3 ±2.2,P =0.000) ; the expressions of c-Myc,Survivin were 7715 ± 2229,4594 ± 1240,which were significantly higher than those in control group (16870 ± 2446,15208 ± 1903,P =0.000) ; the expressions of p53,Bax,Bcl-2,Caspase-1,Fas were not significantly different from those in control group.After 50,100 μg/ml MMI-166 treatment,the expression of c-Myc was significantly down-regulated (0.098 ± 0.003,0.073 ± 0.008 vs.0.169 ± 0.007,F =189.361,P ＜ 0.05) ; and the expression of Survivin was not significantly changed.Conclusions MMI-166 may induce cell apoptosis of SW1990 by down-regulating the expression of c-Myc.

ObjectiveTo investigate the effects of MMI-166 on the proliferation and apoptosis of human pancreatic cancer SW1990 cells.MethodsMMI-166 of different concentrations (25,50,100 μg/ml) were used to treat human pancreatic cancer SW1990 cell for 24,48 h.Effect of MMI-166 on cell proliferation wasdetected by 3- (4,5-dimethyl-2-thiazole) -2-5-biphenly-tetrazole bromide ( MTT ) method and effect on cell apoptosis was tested by Annexin V-PI method and flow cytometry (FCM).ResultsTwenty-four hours after MMI-166 treatment of different concentrations (25,50,100 μg/ml),the inhibitory rates of the cells were (34.23±3.87)％,(44.81 ±2.01)％,(53.91 ±1.74)％,and the corresponding values were (39.95 ± 1.83) ％,( 52.26 ± 3.46 ) ％,( 63.20 ± 2.48 ) ％ at 48 h,which suggested a time-and concentrationdependent manner.The cell's apoptosis rates were (11.19 ±0.47)％,(23.01 ±0.53)％,(28.10 ± 0.52) ％ at 24 h,and the corresponding values were ( 11.19 ± 0.47 ) ％,( 23.01 ± 0.53 ) ％,( 28.10 ± 0.52)％ at 48 h,which were significantly higher than those in control group [ (0.09 ±0.12)％,P ＜0.05].ConclusionsMMI-166 can inhibit proliferation and induce apoptosis of human pancreatic SW1990 cell in a time- and concentration-dependent manner.

Objective To study the expressions of survivin and PTEN in ampullary carcinoma and their clinical significance.Methods The expressions of Survivin and PTEN proteins were tested by EnVision immunohistochemistry in 40 cases of ampullary carcinomas and 8 cases of normal ampulla of vater as control.Results The positive rate of Survivin in ampullary carcinomas was significantly higher than normal human controls (82.5％ vs 0％,P ＜ 0.01 ).The differences of the expression of Survivin in ampullaty carcinoma of duodenal invasion,pancreatic invasion,and lymph node metastasis were significant ( P ＜0.05 ).Significant differences were also observed in the expression rate of PTEN hetween the patients with ampullary carcinoma and the normal controls (50％ vs 100％,P ＜ 0.01 ).The differences of the expression of PTEN in ampullary carcinoma of duodenal invasion,pancreatic invasion,and lymph node metastasis were significant ( P ＜ 0.05 ).Significantly negative correlation was found between the expression of Survivin and PTEN by using spearman correlation analysis ( r =-0.57,P ＜ 0.01 ).Conclusions The abnormal expression of Survivin and PTEN gene may promote tumor genesis and progression of ampullary carcinomas and it may be used as the marker for prognosis.

Objective To investigate of the MMI-166 on the expression of MMP-2,MMP-9 and the cell apoptosis of nude mouse xenografts of SW1990 human pancreatic cancer cells.Methods Establishment of control and experimental groups,randomly,the human pancreatic cancer xenograft model of SW1990 was constructed.The control group was treated with normal saline,and experimental group was treated with MML-166 (200 mg · kg-1 · d-1).The tumor volume and tumor inhibition rate was measured by vernier caliper through length and short diameter.The expression of MMP-2 and MMP-9 protein was observed using immunohistochemistry in the tumor tissues.Apoptosis index was detected by deoxynucleotidyl transferase-mediated nick end labeling (TUNEL method).Results The tumor volume of MMI-166 group (1252.30± 464.84) mm3 was less than the control group (2241.82±208.06) mm3,significantly.The inhibition rate was 34.47％ between the experimental groups (treat with MMI-166) (1.42±0.15) g and control group (2.17±0.20) g.The expression of MMP-2 (2.80 ± 1.10) ％ and MMP-9 (2.60 ± 1.52) ％ protein was significantly downregulated in MMI-166 group,compared with the control group.Apoptotic index in the experimental group (75.60±9.71) ％ was higher than the control group (17.40 ± 10.14) ％,significantly.Conclusion The mechanism of MMI-166 inhibiting pancreatic tumor growth and inducing apoptosis may be related to the suppression of MMP-2 and MMP-9 protein expression.

Objective To study the expression of Survivin and COX-2 in ampullary carcinoma and their clinical significance.MethodsThe expression of Survivin and COX-2 proteins were tested by EnVision immunohistochemistry in 40 cases of ampullary carcinomas,and 8 cases of normal ampulla of vater as the controls.ResultsThe positive rate of Survivin in ampullary carcinonas was significantly higher than that of the controls(82.5％ vs 0,P ＜ 0.01 ). The expression of Survivin in ampullary carcinoma was correlated with duodenal invasion,pancreatic invasion and lymph node metastasis ( P ＜ 0.05 ).Significant difference was also observed in the expression rate of COX-2 between the patients with ampullary carcinoma and the normal controls (67.5％ vs 0,P ＜ 0.01 ).The expression of COX-2 in ampullary carcinoma was correlated with duodenal invasion,pancreatic invasion and lymph node metastasis (P ＜ 0.05). Significantly positive correlation was found between the expression of Survivin and COX-2 by using spearman correlation analysis ( r =0.383,P =0.015).ConclusionThe specific up-regulation of COX-2 gene and Survivin gene may play an important role in the genesis and development of ampullary carcinoma.COX-2 and Survivin may be used as early diagnosis markers and potential therapeutic targets in ampullary carcinoma.