【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 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.
【Abstract】Objective To investigate the imaging features of malignant invasion of major intrahepatic ductal structures (the portal and hepatic venous vasculature, the bilie duct) by primary hepatocellular carcinoma (HCC) using multidetector-row spiral CT (MDCT). Methods We retrospectively analyzed 68 documented HCC patients with tumorous invasion of the major intrahepatic ductal structures who had undergone contrast-enhanced dual-phase MDCT scanning of the upper abdomen.The morphological changes of the portal and hepatic venous vasculature, the bile duct, and the liver parenchyma at both the hepatic arterial phase and portal venous phase images were carefully observed and recorded. Results Among the 68 patients, 47 patients had malignant invasion of the intrahepatic portal venous vessels with secondary tumor thrombus formation; 12 patients had tumor involvement of the hepatic veins and intraheptic segment of the inferior vena cava; Tumor invasion of the bile duct was seen in 9 patents. The direct CT signs of tumor invasion of intrahepatic venous vessels included: ①dilatation or enlargement of the involved vein with intraluminal softtissue “filling defect”； ②enhancement of the tumor thrombus at hepatic arterial phase, the so-called “venous arterialization” phenomenon. The indirect CT signs included: ①arterial-venous shunt, ②early and heterogeneous enhancement of the hepatic parenchyma adjacent to HCC focus, ③cavernous transformation of the portal vein. The CT signs suggesting tumor invasion of the bile duct included: ①dilation of the bile ducts near or proximal to HCC lesion, ②soft-tissue nodule or mass inside the bile ducts. Conclusion Invasion of major intrahepatic ductal structures by HCC will present corresponding CT imaging features. Contrast-enhanced MDCT dualphase scanning combined with appropriate image postprocessing techniques can better evaluate the malignant invasion of major intrahepatic ductal structures.
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.
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.
Through dog models of common bile duct obstruction (BDO), the contents of liver superoxide dismutase (SOD) and malondialdehyde(MDA) were measured 2,3,4 and 5 weeks after BDO. Results indicated that the hepatic MDA content was increased 2 weeks after BDO as compared with control group (Ｐ＜0.01), the hepatic SOD content was decreased 3 weeks after BDO (Ｐ＜0.05). When bile duct obstructing, these changed were more serious. The results suggest that liver has little ability to eliminate the superoxide free radicals after BDO, whereas the lipid peroxidation products increase. It may be one of the mechanisms of liver damage after BDO.
【Abstract】ObjectiveTo analyze the factors influencing the prognosis of patients with bile duct carcinoma after resection. MethodsThe clinical data of 120 patients with bile duct carcinoma receiving resection in our hospital from 1980 to 2004 were collected retrospectively and clinicopathologic factors that might influence survival were analysed. A multiple factor analysis was performed through Cox proportional hazard model. ResultsThe overall 1year, 3year and 5year survival rates were 71.7%, 32.5% and 19.2% respectively. The single factor analysis showed that the major significant factors influencing survival of these patients were histological type of the lesions, lymph node metastasis, pancreatic infiltration, duodenal infiltration, resected surgical margin, perineural infiltration, peripheral vascular infiltration and depth of tumor infiltration (P＜0.05). Lymph node metastasis, pancreatic infiltration and perineural infiltration were found to be the the statistically significant factors influencing survival by multiple factor analysis through the Cox model. ConclusionThe most important prognostic factors for bile duct carcinoma after resection were lymph node metastasis, pancreatic infiltration and perineural infiltration.
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.
ObjectiveTo explore the causes of bile duct injury due to laparoscopic cholecystectomy (LC) and the preventive methods. MethodsA total of 18 patients with bile duct injury (with the occurrence rate of 0.4%) after LC between January 2003 and December 2012 were included. The patients included 5 males and 13 females with the age of 29-63 years old[averaging (42.3±3.6) years old]. The clinical data of the patients were retrospectively analyzed. ResultsIn the 18 cases of bile duct injury, 5 cases occur in emergency operation, 13 cases in selective operation. The operators were attending physician in 13 cases, and senior position in the other 5 cases. The reasons of the injury included misjudgment of the cystic duct in 9, duct aberrance in 3, excessive stretch of cystic duct in 2, 2 Mirizzi syndrome withⅠ-type surgical injury in 2, and right liver duct injury because of inappropriate stripping of gallbladder in 1. ConclusionThe operator's experience, severe conglutination and the bile duct aberrance are the chief causes. Preventive methods include strict system of operation accession, careful selection of candidates, timely laparotomy, and paying attention to the accumulation of operation experience and skills.
Objective To prevent bile duct injury, a new anatomy marker, named “common bile duct window” is created. Methods From November 2005 to March 2006, 60 patients who underwent laparoscopic cholecystectomy were researched in this hospital. All data were collected, including: age, gender, course of disease, body mass index (BMI), blood lipid level (triglyceride and cholesterol), the thickness of gallbladder wall and the degree of cholecystitis. The frequency, location and mean size of “common bile duct window” were recorded and calculated. Patients were divided into two groups according to the presence of “common bile duct window”, and the diference of data between two groups was analyzed by using χ2 test or t test. Results “Common bile duct window” was found at the end of hepatoduoduenal ligament with oval-shaped, the mean longitude of “common bile duct window” was (1.20±0.60) cm, and mean width was (0.45±0.30) cm. “Common bile duct window” were found in 81.6% (49/60) of patients. Age, gender, course of disease, BMI, triglyceride and cholesterol were proved to have no relationship with the presence of “common bile duct window” （Pgt;0.05）, but the thickness of gallbladder wall and the degree of cholecystitis affected the presence （P＜0.05）. Conclusion An oval-shaped “common bile duct window” can be found in almost all patients undergoing laparoscopic cholecystectomy. During the operation, the common bile duct can be located easily by the surgeon through “common bile duct window”, thereby to avoid common bile duct injury when the cyst duct was dissected. It is believed that during laparoscopic cholecystectomy the chances of bile duct injuries can be effectively decreased by the presence of “common bile duct window”.