【Abstract】ObjectiveTo explore a rational and effective operative procedure of fresh tissue samples preservation for biliary tract carcinoma, and to establish a tissue bank with high quality, the authors had made a plan to store up the tissue samples of cholangiocarcinoma resected during operaion in deeplow temperature (cryopreservation), which is based on the clinical preponderance in the treatment of biliary tract diseases of the department of hepatobiliary surgery of General Hospital of P.L.A., and will be a good foundation for the systemic basic research of bile duct carcinoma. MethodsCases of biliary tract tumors confirmed by pathology were selected from the inpatients of Department of Hepatobiliary Surgery in General Hospital of P.L.A. from Jan. 2000 to June 2001. Fresh tissues were taken from the excised sample, which were stored in three different disinfectant tubes labelled cancer ( or tumor ) group, peritumor group and normal group. They were stored in liguid nitrogon container temporarily, and transferred into refrigerator for longtime storage as quickly as possible. The slices divided from the sample were grouped according to the dimensional space apart from the margin of tumor. Pathological diagnosis must be made on paraffin embedded samples. A part of the tissues was used to isolate total RNA by Trizol reagent for integrality judgment of it.ResultsTwentyone excised samples were stored including 4 intrahepatic bile duct carcinoma, 6 hepatic hilar cholangiocarcinoma, 7 extrahepatic bile duct carcinoma, and 4 duodenal ampulla carcinoma. The number and size of samples stored and duration of severance were not the same for reason of the difference of tumor volume, difficulty of operation and artificial jamming. The average duration was (47.60±43.87) min. The precipitated total RNA could be seen at the bottom of Eppendorf tube, of which the rate of A260/A280 was calculated about 1.6 to 1.8. It also could be seen in the 1%agarose gel electrophoresis for the obvious two bands of 28s and 18s, in which the28s band might be twofolds lighter than the 18s. Conclusion It is an important basic work for research of genes related to human disease to built a sample bank of human genetic resource. The present program for bile duct carcinoma tissue severance and storage is feasible and could supply the goodquality sample for further study. It must be reminded that the informed consent is needed and the left sample should be sufficient for postoperative pathological examination before the performance, which should be done by a fixed and experienced researcher group. Limitted to the operation of bile duct carcinoma, the time for beginning tissue severing should be in one hour after the sample excision, and samples should be divided into slices in short time to avoid decomposition of component during the following schedule.
ObjectiveTo summarize the clinical experience of 180 general surgeries under da Vinci surgical system. MethodsFrom January 2009 to October 2010, 180 patients with hepatopancreaticobiliary and gastrointestinal disease underwent robotic surgeries by using da Vinci surgical system. The case distribution, intra-and post-operative data were analyzed. ResultsA total of 171 patients had underwent total robotic surgeries and nine patients converted to hand-assisted procedure (5.0%, 9/180). The surgery for hepatic portal was performed in 63 cases of patients, including surgery for hilar cholangiocarcinoma in 36 cases, gallbladder carcinoma in 10 cases, complex calculus of intrahepatic duct in 12 cases, and iatrogenic biliary duct injury in 5 cases. The pancreatic surgery was performed in 44 cases of patients, including pancreatoduodenectomy in 16 cases, distal pancreatectomy in 6 cases, medial pancreatectomy in 1 case, pancreatic cyst-jejunum anastomosis in 1 case, and palliative surgery in 20 cases. Hepatic surgery was in 19 cases and gastrointestinal surgery in 12 cases of patients. Other procedures were in 42 cases of patients, including common bile duct exploration and removing the stone, retroperitoneal lymph nodes dissection, and splenectomy, etc. All of ten cases of patients with obstructive suppurative cholangitis received emergency surgery by the robot and postoperative infection symptoms were controlled, and shock was corrected quickly. No death occurred during the perioperative period. Postoperative complications occurred in 12 cases (6.7%, 12/180) and 2 cases died (1.1%, 2/180). Conclusionsda Vinci surgical system can carry out all kinds of general surgery, especially complicated and difficult hepatobiliary and pancreatic surgery, which improves the development of minimally invasive surgery.
ObjectiveTo summarize the clinical experience of 58 operations for biliary malignant tumor with da Vinci surgical system. MethodsFrom January 2009 to October 2010, 180 patients with hepatopancreaticobiliary and gastrointestinal disease underwent robotic surgeries by using da Vinci surgical system, including 58 patients with biliary malignant tumor. The case distribution, intra and postoperative data were analyzed. ResultsOf 58 patients, 3 patients with intrahepatic bile duct cystadenocarcinoma received wedge resections of liver. In 36 patients with hilar cholangiocarcinoma, anatomical left hemihepatectomies were performed in 3 cases, resection of extrahepatic duct and gallbladder bridge type biliary revascularization in 3 cases, resection of extrahepatic duct and biliary-enteric Roux-en-Y anastomosis in 14 cases, tumor resection and revascularization of hepatic portal bile duct in 1 case, palliative external drainage of intrahepatic bile duct in 5 cases, and Y-internal drainage of hepatic portal in 10 cases. In 10 patients with gallbladder carcinoma, resection of extrahepatic duct and gallbladder and biliaryenteric Roux-en-Y anastomosis in 2 cases, cholecystectmy in 3 cases, cholecystectmy and external drainage of intrahepatic bile duct in 1 case, cholecystectmy and Y-internal drainage by suspension of hepatic portal in 4 cases. A patient with middle bile duct cancer received radical resection of cholangiocarcinoma and biliary-enteric Roux-en-Y anastomosis. Of 8 patients with distal bile duct cancer, Whipple procedure were performed. Of 58 patients, 2 cases converted to hand-assistant procedure (3.4%). For all patients, operation time was (6.18±1.71) h, blood loss was (116.66±56.06) ml, blood transfusion was (85.55±38.28) ml, ambulation time was (9.10±2.91) h, feeding time was (14.95±4.35) h, and hospital stay was (12.81±4.29) d. Postoperative complications occurred in 8 cases (13.8%), including bile leakage (3 cases), wound bleeding (1 case), pancreatoenteric anastomotic leakage (2 cases), pulmonary infection (1 case), and renal failure (1 case). Of these 8 cases, 6 cases recovered smoothly and 2 cases die of severe pulmonary infection and renal failure after conservative treatment (3 or 4 weeks), therefore, the mortality of patients was 3.4%. In 36 patients with hilar cholangiocarcinoma, 19 cases died (on 2 monthes 4 cases, on 6 monthes 5 cases, on 10 monthes 8 cases, and on 12 monthes 2 cases after operation), 11 cases survival well (gt;26 monthes 4 cases, gt;22 monthes 3 cases, and gt;19 monthes 4 cases), and 6 cases required hospitalization. Of 10 patients with gallbladder carcinoma, 7 cases died (on 3 monthes 1 case, on 5 monthes 1 case, on 8 monthes 1 case, on 11 monthes 3 cases, and on 12 monthes 1 case after operation) and 3 cases survival (gt;17 monthes 2 cases, gt;13 monthes 1 case). In 8 cases undergoing pancreatoduodenectomy, 5 cases died (on 4 monthes 2 cases, on 6 monthes 2 cases, and 10 monthes 1 case after operation) and 3 cases survived well over 2 years. Three patients with intrahepatic bile duct cystadenocarcinoma survived over 1 year. Conclusionsda Vinci surgical system can carry out all kinds of surgery for biliary malignant tumor, especially prominent in the complicated surgeries for hepatic portal, which breaks through the restricted area of laparoscope in hepatobiliary malignant tumor.
Objective To obtain the full-length gene and functional domains of FXYD6 gene which is a cholangiocarcinoma related gene. Methods A new strategy with the integration of bioinformatics and molecular biology was used. Bioinformatical methods were used to analyze the full-length sequence, and to predict the functional domains of its protein. And the full-length sequence of FXYD6 was isolated by polymerase chain reaction from fetal hepatic, brain and spleen cDNA libraries, and then cloned in pGEM-T vector for sequence analyzing. Goldkey Sequence Analyzing Software was used to analyze the sequence of candidate domain without signal peptide.Results The full-length sequence of FXYD6 was isolated by Touch-down PCR from fetal hepatic and brain cDNA library, but was not from spleen cDNA library. The open reading frame Finder software was used in the National Center for Biotechnology Information website to find the most probable encoding regions of FXYD6 gene. And the +1 phase was selected as the template sequence, from 67 bp to 354 bp, to predict the functional domains by Goldkey Sequence Analyzing Software. The signal peptide was located from 1 amino acid (aa) to 17 aa， and the main domain was composed from 18 aa to 34 aa. The region between 35 aa and 57 aa was the transmembrane region. The FHYD peptide chain was highly conserved amino acids. Conclusion The study of full-length cDNA cloning of FXYD6 gene and its functional domains provides the basis for understanding the relationship between the structure and function of FXYD6. More work shall be performed on FXYD6 protein and its influence on the mechanism of cholangiocarcinoma.