Objective To constuct and identify a specific siRNA targeting mucins (MUC1) expression vector.Methods A pair of oligonucleotide completing and coding hairpin MUC1-siRNA synthesized were inserted into pGC silencerTM U6/Neo/RNAi vector.Double enzyme digestion,PCR test and DNA sequencing were used to identify the recombinant plasmid.Then,special MUC1-siRNA was transfected into cholangiocarcinoma cells with Lipofectamine TM 2000.In vitro,MUC1 mRNA and protein expression was tested by RT-PCR and Western Blot respectively.Results Double enzyme digestion,PCR test and DNA sequencing confirmed that the MUC1 specific siRNA expression vector was constructed successfully.After transfection,the expression of MUC1 mRNA and protein in QBC939 (experimental group) decreased significantly (P

Objective:To understand the levels of selenium and T-2 toxin in the historical critical disease area of Kashin-Beck disease in Aba State of Sichuan Province.Methods:From July 2018 to February 2019, in five cities (counties) of historical critical Kashin-Beck disease areas, namely, Ma'erkang, Rangtang, Aba, Ruo'ergai and Hongyuan, one township was selected, respectively, and then one village in the historical critical disease area and one village in non-disease area were selected from the selected townships. Hair samples from 30 children aged 6 to 12 years old were collected to test selenium content in each village. Staple food samples of 10 households were collected to test the selenium and T-2 toxin contents in each village. Ten soil samples were collected from each village to test the selenium content.Results:The medians selenium of children's hair in the diseased and non-diseased villages ( n= 180, 120) were 0.260 0 and 0.330 0 mg/kg, respectively, and the diseased village was lower than non-diseased village ( Z = - 6.319, P < 0.01). In Ma'erkang and Hongyuan, the medians selenium in the diseased villages were lower than those in non-diseased villages ( Z = - 3.327, - 4.939, P < 0.01), and there were no statistically significant differences between the diseased villages and non-diseased villages in Rangtang and Ruo'ergai ( P > 0.05). The medians selenium of staple food in the diseased and non-diseased villages ( n = 110, 70) were 0.005 8 and 0.018 0 mg/kg, respectively, and the diseased village was lower than non-diseased village ( Z = - 2.263, P < 0.05). In Ruo'ergai, the median selenium in the diseased village was lower than that in non-diseased village ( Z = - 2.306, P < 0.05), and there were no statistically significant differences between the diseased villages and non-diseased villages in Ma'erkang, Rangtang and Hongyuan ( P > 0.05). The T-2 toxin contents in staple food were all less than detection limit (1 μg/kg) in the diseased and non-diseased villages ( n = 103, 65). The soil selenium contents in the diseased and non-diseased villages ( n = 60, 40) were (0.045 3 ± 0.021 5) and (0.065 8 ± 0.045 4) mg/kg, respectively, and the diseased village was lower than non-diseased village ( t = 2.672, P < 0.05). The soil selenium content in the diseased village was lower than that in non-diseased village in Hongyuan ( t = 14.838, P < 0.01). There were no statistically significant differences in the soil selenium content between the diseased villages and non-diseased villages in Ma'erkang, Rangtang and Ruo'ergai ( P > 0.05). Conclusions:The overall selenium level of children's hair is at a medium level (0.25 - < 0.50 mg/kg) in the historical critical disease area of Aba State of Sichuan Province, the selenium levels of staple food and soil are at a deficiency level (< 0.025, < 0.125 mg/kg), and the diseased villages are lower than non-diseased villages. No T-2 toxin is detected in staple food.

Objective To investigate the effect of c-myc ASODN on the proliferation and invasion of human bile duct carcinoma cell line QBC939. Methods QBC939 cells was conventionally cultured. C-myc ASODN was designed and transfected into QBC939 cell line. MTT assay and transwell experiment were used to study cell proliferation and invasion of QBC939 cells. Results MTT assay showed that cell survival rate in ASODN group was significantly lower than that in blank group(P < 0.05). Transwell experiment showed that the num-ber of cells penetrated in ASODN group was significantly lower than that in blank group(P<0.01). The cell survival rate and the number of cells penetrated in vechicle group had no difference with blank comparison group(P>0.05). Conclusions C- myc ASODN can inhibit the proliferation and invasion of QBC939 cells.

Objective To investigate therapeutic effect and mechanism of hyperbaric oxygen and ulinastatin respectively or combinatively used to treat acute necrotizing pancreatitis (ANP). Methods One hundred and twenty SD rats were divided into 6 groups randomly: group of normal control, group receiving sham operation, group of untreated acute necrotizing pancreatitis (ANP group), group of acute necrotizing pancreatitis treated with hyperbaric oxygen (HBO group), group of acute necrotizing pancreatitis treated with ulinastatin (ULT group), and group of acute necrotizing pancreatitis treated with combined hyperbaric oxygen and ulinastatin (HBO+ULT group). The rat model of acute necrotizing pancreatitis was established according to Aho HJ et al. Concentrations of amylase, TNF?, TXB-2 and 6-keto-PGF- 1? in blood were measured through ELISA or radioimmunoassay. Changes of pancreatic histopathology were investigated. SPSS 10.0 was used in statistical analysis. Results The concentrations of amylase, TNF?, TXB-2 in the ANP-treated groups were significantly lower than those of ANP group (P

Objective To explore the reasons,diagnosis and treatment of residual cholecystitis(RCC) with gallstones. Methods The clinical data of 36 RCC patients with gallstones identified by operation were retrospectively analyzed. Results All the 36 patients were cured by reoperation. Residual cholecystectomy was performed on 8 patients, and residual cholecystecomy plus common bile duct exploration and T tube drainage on 28 patients. Thirty one patients were followed up for 3 months to 12 years,93.55% of the patients had good results. Conclusions The main reason of residual cholecystitis with gallstones was not followed the principle of "identify cut identify" during cholecystectomy .The clinical presentation of RCC is similar to that of cholecystitis with gallstones .The accurate rate of auxiliary examinations is low,so the results of these exammations should be analyzed comprehensivly in the diagnosis. The principle of "identify cut identify" should be followed during the reoperation. The common bile duct and common hepatic duct should be opened first and then the residual gall be resected.

Objective To study the operative procedure for stage III and IV hilar cholangiocarcinoma. Methods A crescent shape excision on the edges of multiple hepatic segments followed by a 'skirt edge′ form hepatoenterostomy to drain the multiple hepatic ducts was used to treat unresectable stage III,IV hilar cholangiocarcinoma, . Results (1)the mean survival time was 15.65 months;(2)the patient comfortable index was 81.5%;(4)there was no operative death in the series. Conclusions The 'skirt edge' form hepatoenterostomy is a feasible and effective palliative method for unresectable stage III,IV cholangiocarcinoma.

Objective To investigate the application of selective hepatopetal blood occlusion techniques in anatomic hepatectomy.Methods We retrospectively reviewed the clinical data of 259 patients with hepatolithiasis or liver tumor undergoing anatomic hepatectomy under selective hepatopetal blood occlusion from January 2006 to December 2009.Results Totally,183 cases with hepatolithiasis and 76 cases with liver tumor underwent anatomic hepatectomy under selective hepatopetal blood occlusion.The average intra-operation blood loss was 210 mL(120-1 600 mL);post-operation incidence of complications and the rate of residual stones was 10.9% and 4.2%,respectively.Thre was no operative death in this series.The intrahepatic recurrence and metastasis rate of liver tumor was 23.6% and the median recurrence was 16.3 months.Conclusions The use of a appropriate selective hepatopetal blood occlusion during anatomic hepatectomy for hepatolithiasis and liver tumors is an effective measure to reduce surgical complications and improve outcome.

Objective To investigate the efficacy of duodenum-preserving pancreatic head resection (DPPHR) for the treatment of chronic pancreatitis combined with type Ⅰ and Ⅲ pancreatic duct stones.Methods The clinical data of 55 patients with chronic pancreatitis and type Ⅰ and Ⅲ pancreatic duct stones who were admitted to the People's Hospital of Hunan Province from June 2008 to June 2013 were prospectively analyzed.All the patients were randomly divided into the pancreatoduodenectomy (PD) group (27 patients) and the DPPHR group (28 patients).There were 18 patients with chronic pancreatitis and type Ⅰ pancreatic duct stones and 9 patients with chronic pancreatitis and type Ⅲ pancreatic duct stones in the PD group.There were 16 patients with chronic pancreatitis and type Ⅰ pancreatic duct stones and 12 patients with chronic pancreatitis and type Ⅲ pancreatic duct stones in the DPPHR group.Patients in the PD group received PD + Child anastomosis + end-toside pancreato jejunal anastomosis + pancreatic stent placement + end-to-side cholangiojejunostomy.Patients in the DPPHR group received free of duodenum + pancreatic duct incision + resection of pancreas at 1 cm ahead of the pancreatic duct + extraction of the pancreatic duct stones + pancreaticoduodenal Roux-en-Y anastomosis.Patients were followed up via out-patient examination till December 2013.The measurement data were analyzed using the t test or Mann-Whitney U test,and the count data were analyzed using the chi-square test.Results During the operation,2 patients in the PD group were converted to the DPPHR group and 1 patient in the DPPHR group was converted to the PD group.No patient died during the perioperative period,and the symptoms including abdominal pain and diarrhea were alleviated at postoperative week 2.The operation time,blood loss,duration of postoperative hospital stay,total expenses and incidence of complications were (7.5 ± 1.6) hours,(460 ± 88) mL,(18.0 ± 3.5) days,(7.8 ± 2.1) × 104 yuan,19.2％ (5/26) in the PD group,and (4.0 ± 1.0) hours,(120 ± 36) mL,(9.5 ± 2.9) days,(3.9 ± 1.2) × 104 yuan,3.4％ (1/29) in the DPPHR group,there were no significant differences in the operation time,blood loss,duration of hospital stay,total expenses and incidence of complications between the 2 groups (t =9.358,11.365,6.325,8.647,x2 =3.976,P ＜ 0.05).Fifty-three patients were followed up,with the median time of 33 months (range,6 months to 5 years).No patient died during the follow-up.Twenty-four patients in the PD group were followed up,2 patients had slight abdominal pain,1 patient had severe abdominal pain due to pancreatic duct stenosis,and the symptom was alleviated after resection of partial pancreas ; the condition of 12 patients was improved among the 19 patients with diabetes.Twenty-nine patients in the DPPHR group were followed up,2 patients had slight pain; the condition of 16 patients were improved among the 22 patients with diabetes.Conclusion DPPHR is an ideal surgical procedure for patients with chronic pancreatitis and type Ⅰ and Ⅲ pancreatic duct stones.

Objective To investigate the operative techniques for refractory cholelithiasis. Methods The clinical data of 521 patients with refractory cholelithiasis who had been admitted to People's Hospital of Hunan Province from January 1990 to December 2007 were retrospectively analyzed. Results All patients received surgery. After the operation, 3 patients died of liver and kidney failure, the remaining 518 patients were cured without severe complications. The results of B ultrasound and computed tomography showed residual stones in 78 patients (15.1%). Four hundred and twenty-three patients (81.7%) were followed up for 5 months to 17 years (mean, 7.5 years), and the rate of positive effect was 90.1% (381/423). Conclusions Most of the refractory cholelithiasis can be cured radically. Individualized surgical planning, fine and standard surgical procedure are key to the treatment effect.

Objective To summarize the experience in surgical management of bile duct injury. Methods The clinical data of 683 patients with bile duct injury who were admitted to the Hunan People's Hospital from August 1990 to December 2008 were retrospectively analyzed. Of all the patients, seven received hepatectomy +T tube drainage, two received liver repair + T tube drainage, four received external biliary drainage + hepatectomy,27 received liver repair or hepatectomy + silicone support, 233 received temporary portal triad clamping + gelatin sponge hemostasis, 72 received biliary repair + T tube drainage, 248 received hepatobiliary basin Roux-en-Y anastomosis, 22 received external biliary drainage, 61 received long arm T tube drainage, two received pancreaticoduodenectomy and five received hepatectomy + T tube drainage. The surgical outcomes were evaluated by analyzing the results of the follow-up. Results The surgical outcomes were ranked excellent, good and poor according to the condition of patients and the results of imaging examination. Six hundred and twelve patients were followed up for 8 months to 19 years, and the surgical outcomes were excellent in 337 patients (55.1%), good in 214 patients (35.0%) and poor in 61 patients ( 10.0% ). Conclusion The surgical outcome of bile duct injury could be satisfactory if the approach of the surgery is properly selected.