ABSTRACT
Objective To observe the effects of interfering Twist expression on glycolysis in pancreatic cancer PANC1 cells and explore the potential mechanism.Methods Twist targeting siRNA (siTWIST) was designed and synthesized,and pancreatic cancer PANC1 cells were transfected with siTWIST using liposome,with nonspecific siRNA (siNC) transfected cells as negative control group and untransfected cells as blank control group.RT-PCR and Western blot were used to detect the expression levels of Twist mRNA and protein,and Akt and p-Akt protein expression in different groups;cell proliferation was detected by MTT;and glycolysis key enzyme (pyruvate kinase and hexokinase) activity and the content of lactic acid in the supernatant of culture fluid were detected.Results Twist mRNA level in control group,siRNA group and siTwist group expression was 1.00 ± 0.09,1.01 ± 0.08 and 0.36 ± 0.02;Twist protein level was 0.41 ± 0.05,0.42 ± 0.04 and 0.12 ± 0.06;the mRNA and protein expression in siTWIST group was obviously lower than those in control group and the difference was statistically significant (P ＜ 0.05).The cell survival rate was (100.02± 9.36)％,(100.01 ± 10.25)％ and (59.32± 7.26)％,which in siTWIST group was obviously lower than that in control group;pyruvate kinase activity was (0.73 ± 0.05)U/mg,(0.72 ± 0.08) U/mg and (0.43 ± 0.05) U/mg;the activities of hexokinase were (4.98 ± 0.48) U/mg,(4.96 ± 0.52) U/mg and (2.54 ± 0.21) U/mg;the content of lactic acid was (20.36 ± 2.61) mmol/L,(20.64 ± 3.05) mmol/L and (9.48 ± 1.09) mmol/L;the ratio of p-Akt/Akt was 0.65 ± 0.04,0.63 ± 0.07,and 0.25 ± 0.04,which in siTWIST group was obviously lower than that in control group.Conclusions Interference with Twist expression could inhibit the proliferation and glycolysis pathway in pancreatic cancer cells,and the mechanism may be related to the Akt signaling pathway.
ABSTRACT
Objective To explore the factors which could lead to a preoperative diagnosis and to guide the management of cholecystoduodenal fistula (CDF).Method The experience on the diagnosis and treatment of 22 patients with CDF were retrospectively studied.These patients came from 1242 patients who received biliary tract operation in our hospital from 2001 to 2012.Results 64％ (14 of 22 patients) had 3 out of 5 of the following symptoms/signs:(1) symptoms of recurrent chills and fever,with no or only mild jaundice; (2) significant atrophy or disappearance of gallbladder on computed tomography (CT) ; (3) CT revealed complex anatomy in right upper abdomen with dilated loops of bowel; (4) ultrasound or CT revealed pneumobilia or pneumo-gallbladder; (5) barium study or duodenal endoscopy revealed obvious deformation in duodenal bulb or abnormal opening.There was no perioperative death.Morbidities included biliary fistula which presented on postoperative day 6 and day 7 in 2 patients,respectively.The daily volume of bile drainage was about 500 ml,and the biliary fistula healed after a month of conservative treatment.In addition,there were 6 patients who had infected wound,8 patients with right pleural effusion,and 8 patients with residual calculi.There was no intestinal fistula or biliary stricture.Conclusions Careful preoperative history taking and CT/uhrasound studies significantly improved the diagnostic rate of CDF.Individualized treatment reduced complications and improved clinical results.