Objective To investigate the clinical significance of routine application of choledochoscope during operation of biliary duct. Methods The clinical data of 136 patients with the routine application of intraoperative choledochoscope dealing with bile duct diseases in this hospital from October 2003 to July 2009 were analyzed and summarized. Results Intraoperative choledochoscope inspection, taking stones and targeted surgery were performed in 116 cases with extrahepatic and (or) intrahepatic bile duct stones. The taking out rate of extrahepatic bile duct stones was 100% (85/85), the residual stone rates of extrahepatic and intrahepatic bile duct were 0 (0/85) and 22.6% (7/31), respectively, with the total residual stone rate was 6.0% (7/116). By using intraoperative choledochoscope, benign intrahepatic bile duct stricture was found in 13 cases, malignant extrahepatic and intrahepatic bile duct stricture in 8 cases. Four cases of hilar cholangiocarcinoma and 4 cases of common bile duct cancer, 2 cases of lower segment of common bile duct polyp, 3 cases of hepatolithiasis with the left hepatic bile duct carcinoma were diagnosed by biopsy via choledochoscope. Causes were confirmed by applying choledochoscope in 16 patients with obstructive jaundice. The use of choledochoscope with surgical treatment enabled benign and malignant bile duct stricture to achieve good results, without serious complications such as bile duct dilaceration, subphrenic abscess or acute cholangitis. Conclusions Using choledochoscope can tremendously reduce the residual rate of stone in biliary duct surgery, increase the definite diagnosis rate of biliary duct diseases and play a role of reasonable instruction in its treatment options. It has unique advantages in identifying causes of jaundice. It is remarkable that the role of applying choledochoscope to diagnose and cure biliary duct diseases. It should be widely used.
Objective To investigate the phenotypic change and proliferation of fibroblasts in human inflammatory strictured bile duct wall. Methods We observed the density and ultrastructure of fibroblasts, and the histologic structure in human normal bile duct wall and inflammatory strictured bile duct wall by light and electron microscope.Results The results showed that fibroblasts were the main source of extracellular matrix production in bile duct wall. The phenotype of fibroblasts in inflammatory strictured bile duct wall changed obviously, quiescent fibroblasts were activated and transformed to myofibroblasts, with massive proliferation. Conclusion These data suggest that massive proliferation of activated fibroblasts and myofibroblasts is the main source of extracellular matrix overproduction which results in inflammatory bile duct stricture.
Immunohistochemical study on 39 specimens of hepatobilibary duct stricture due to stones were performed. Collagen types Ⅲ and Ⅳ were studied by quantitative analysis. The results showed that significant increase of type Ⅲ collagen was found in the stenotic bile duct wall, the portal area and liver sinusoid with fibrosis. Abnormal increasing of type Ⅳ collagen was found in the liver sinusoid of the stenotic bile duct.
【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.
Objective To observe the influence of connexin 43 (Cx43) on bystander effects induced by cytosine deaminase (CD) and herpes simplex virus thymidine kinase (HSV-tk) coexpression suicide genes system in human cholangiocarcinoma QBC939 cells and transplantation tumors in nude mice. Methods In vitro, the CD+tk+ and CD+tk+Cx+ cells were respectively treated with 5-fluorocytosine (5-FC) and ganciclovir (GCV). The cytotoxic efficacy was evaluated by microculture tetrajolium test (MTT) method. In order to investigate the influence of Cx43 on bystander effects, the volumes of transplantation tumors of the CD+tk+ and CD+tk+Cx+ cells were measured before and after application of 5-FC and GCV. Results CD and tk gene were stably expressed in transfected QBC939 cells. Increasing expression of Cx43 was determined by testing for the presence of Cx43 mRNA by RT-PCR and the presence of Cx43 protein by Western blot in CD+tk+Cx+ cells. The killing effect of 5-FC and GCV on CD+tk+Cx+ cells was more effective than that on CD+tk+ cells both in vitro and in vivo. Conclusion Double suicide genes system CD/5-FC+tk/GCV could induce remarkable killing effect on cholangiocarcinoma cells in vitro and transplantation tumors in vivo. The cotransfection of Cx43 gene is able to enhance the bystander effects and the inhibition of carcinoma cells.
OBJECTIVE: To explore the possibility of repair of the extensive bile duct injuries with expanded polytetrafluoroethylene (ePTFE). METHODS: A total of 36 local healthy hybrid dogs were employed to establish bile duct injury models by means of partial removal of bile duct (group A, 14 dogs), excision of a segmental duct (group B, 10 dogs), and ligation of the lower part of common bile duct(group C, 5 dogs; group D, 7 dogs). Group A were patched with ePTFE mesh, group B replaced by ePTFE tube. Interposition of the grafts between gallbladder and duedenum or jejunum was performed on group C and group D. The animals’ postoperative performance status were evaluated. Cholangiography was used to define the patency of bile ducts. Tissues of bile ducts and liver were taken at 3 days, 2, 4, 8, 12 and 52 weeks for microscopic and ultrastructural examination to observe the healing process of bile duct and morphological changes in the liver. RESULTS: Group A with ePTFE patch covered by epithelium had a high patency rate of 75%(9/12) and pathological damages were not found in the liver. The patency rate of group B was merely 40%(4/10), to some extent, accompanying damages in the liver. The grafts of group C and group D were fully expelled, ultimately leading to cystic-duodenal or cystic-jejunal fistulas formation. CONCLUSION: The study suggests that application of ePTFE patch to repair bile duct defects is feasible.
Objective To determine whether local delivery of c-myc shRNA could inhibit hyperplasia and lithogenic potentiality in a rat model of chronic proliferative cholangitis (CPC) via specific blockade of the c-myc expression. Methods The CPC animal model (CPC group) was established via retrograde insertion of a 5-0 nylon thread into the common bile duct through Vater’s papilla. Three kinds of c-myc shRNAs were then respectively injected in c-myc shRNA group, which were included shRNA-1, shRNA-2, and shRNA-3, respectively. Negative control group and sham operation group were established for comparison. Subsequently, histopathological changes of bile duct wall were observed by HE, Massion, and PAS/AB staining; c-myc protein was detected by immunohistochemistry method; 5-bromodeoxyuridine (BrdU) protein was tested by immumofluorescence method; c-myc, Mucin 3, and Procollagen Ⅰ mRNAs were detected by real time PCR; Ki-67 protein was determined by Western blot; Activity of β-glucuronidase was measured by modified Fisherman method. Results ①Compared with the CPC and negative control groups, biliary tract mucosa epithelium (HE staining), submucosal acid mucinous gland (mid-blue staining, PAS/AB staining), and degree of over-hyperplasia of collagen fiber in bile duct wall (blue staining, Massion staining) were weaker in the c-myc shRNA group. ②The expressions of c-myc mRNA, Mucin 3 mRNA, Procollagen Ⅰ mRNA, Ki-67 protein, and β-G activity in the c-myc shRNA group were lower than those of the CPC and negative control groups (Plt;0.05), but higher than those of the sham operation group (Plt;0.05). Conclusion c-myc shRNA treatment could effectively inhibit the hyperplastic behavior and lithogenic potential of CPC, which might help to prevent the biliary restenosis and stone recurrence.
Eighty two cases of acute gallstone pancreatitis on early operation are reported and the significance of the clinical picture and pathology are analysed. The data showed that gallstone was found in 85.5%, among the cases of them mulliple gallstone was 71.1%, dilated cystic duct was 26.4%, common bile duct stone 36.8%, distal bile ductal stricture was found in 9.3%, and anomalous conjunction of biliary and pancreatic duct was 20.1%. Sixteen cases with serious pancreatitis were determined on operation, but death rate was 3.7% only. The authors claim that early operation may be of value in patients of acute gallstone pancreatitis with or without jaundice espesially in bile duct obstruction.
ObjectiveTo explore the possibility of intrahepatic biliary perfusion by contrast enhanced ultrasonography via bile duct.
MethodsSixteen rabbits weighing 2 000-2 500 g were studied. A 22 gauge catheter was inserted in the common bile duct by surgery. The rabbits were divided into 4 groups according to the concentration of contrast agent, including group A:1/100 of standard concentration; group B:1/200 of standard concentration; group C:1/400 of standard concentration; and group D:1/800 of standard concentration. We observed the filling status, presence of outflow of contrast enhanced signal and satisfactory enhanced time after the injection of contrast agent via the catheter in common bile duct.
ResultsFive milliliter of contrast agent was injected in each rabbit. The number of homogeneous filling in group A, B, C and D was 4, 4, 3 and 0 respectively. The number of presence of outflow of contrast enhanced signal in group A, B, C and D was 4, 1, 0 and 0 respectively. The satisfactory enhanced time in group A, B, C and D was (340±29) s, (284±37) s, (82±8) s and 0 s respectively. There was no statistical difference in the satisfactory enhanced time between group A and B (P=0.06) while significant difference in the satisfactory enhanced time between group A and C (P < 0.01), and between group B and C (P < 0.01) was found.
ConclusionIt is possible to perfuse the intrahepatic biliary system by sonographic contrast agents via bile duct, and 1/200 of standard concentration is a proper concentration of contrast agent to achieve satisfactory imaging.
Objective To investigate the prevention and treatment strategy of bile duct injury (BDI) in laparoscopic cholecystectomy (LC). MethodsLatest progress was reviewed based on recent documents and the experience on BDI in LC in our department. ResultsWith the popularity of LC, BDI in LC is increasing. The reasons include illegibility and variability of local anatomy in gallbladder trigone,injury caused by galvanothermy, as well as operator’s overconfidence. In order to prevent BDI, we should apply more blunt dissection, not to use electrocogulation if possible and to study local anatomy and its variance clearly. The common bile duct and common hepatic duct should be clearly identified. Intraoperative cholangiography, laparoscopic ultrasonography and hepatobiliary scintigraphy are selections as necessary.The treatment of BDI depends on the type of BDI and its site and local condition.The treatment includes end to end anastomosis, repairing the defect, choledochoduodenostomy, RouxenY choledochojejunostomy and so on. T tube should be maintained in place for more than half a year after operation.Conclusion The key to improve the prognosis of BDI is prevention and treatment in proper time and in correct way.
