【Abstract】ObjectiveTo evaluate the injury of common bile duct in immediate removal of the ligation in cholecystectomy. MethodsEighteen healthy Japanese rabbits were selected and divided into three groups randomly: A group treated with simple cholecystectomy, B group with cholecystectomy plus common bile duct crossligation and C group with cholecystectomy plus hepatic bile duct conjunction “Y”type ligation. The ligation was removed after 5 min in B and C groups. The levels of serum transaminase and bilirubin and pathological changes of bile duct and liver in each group were observed respectively. ResultsThere were no statistic difference in the levels of GPT, GOT, total bilirubin (TB), direct bilirubin (DB) and DB/TB on 12 hours before operation and the 1st and 7th day after operation between A and ligation groups (Pgt;0.05). But there were statistic difference in the those indexes and pathological changes of bile duct and liver between A and ligation groups on the 30th and 90th day after operation (Plt;0.05). ConclusionDuring cholecystectomy, immediate removal of common bile duct ligation doesn’t affect shortterm results, but the long-term results are bile duct stricture and obstruction.
【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.
Objective To observe the expression and distribution of transforming growth factor-β1 (TGF-β1) in the healing process of bile duct and discuss its function and significance in the process of benign biliary stricture formation. Methods An injury to bile duct of dog was made and then repaired. The expression and distribution of TGF-β1 in the tissue at different time of the healing process were studied after operation with immunohistochemical SP staining. Results TGF-β1 staining was observed in the granulation tissue, fibroblasts and endothelial cells of blood vessels. High expression of TGF-β1 was observed in the healing process lasting for a long time. Conclusion The high expression of TGF-β1 is related closely with the fibroblast proliferating activity, extracellular matrix overdeposition and scar proliferation in the healing process of 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 comment the diagnosis and treatment the bile leakage from the injuried abnormal minute biliary in our laparosicopic cholecystectomy (LC) practice. Methods Fourteen cases of minute biliary duct injury in 2 050 cases of LC were studied retrospectively. Among them, 6 cases had been found the points of leakage during operation, and the points were treated by titanium nips. In 4 cases even though the bile leakage could be seen, but the points of leakage could not found, and were treated by drainage. Four cases with peritonitis, 1 needed to be explored, and treated with suture ligature, 1 was explored by laparoscopy again, another two cases were treated with multiple hole catheters to drainage of the abdominal cavities through stab wounds. Results All 14 cases recovered. Conclusion Small bile leakage in LC is almost inevitable. It is the best that the bile leakage can be discovered during operation and to be treated. If it is discovered after operation, an open or laparoscopic exploratory laparotomy and adequate drainage would be needed. In the case of small amount of leakage, catheter drainage through stab wound is feasible.
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.
Objective To investigate the effects of sustained-release basic fibroblast growth factor (bFGF) on healing of bile duct defect. Methods A model of bile duct wall defect (2 cm in length and 1/3-2/3 of the bile duct circumference in width) was made in 24 pigs (male or female, weighing 15-30 kg), and then defect was repaired with sustained-release bFGF collagen membrane (2.0 cm × 1.0 cm × 0.5 cm in size) in the experimental group (n=12) or with collagen membrane (2.0 cm × 1.0 cm × 0.5 cm in size) alone in the blank control group (n=12). Another 4 healthy pigs were used to obtain normal bile duct as normal control group. The survival condition of pigs was observed after operation; at 1, 2, and 3 months after operation, the blood sampling was collected to test the changes of liver function, and the bile duct specimens were harvested to count the microvessel density (MVD) and submucosal gland by HE staining and immunohistochemistry staining; and at 3 months after operation, cholangiography examination was done. Results All the animals survived to completion of the experiment. Intra-abdominal adhesion was serious in the experimental and blank control groups at 1 week after operation, but the adhesion was markedly improved in the experimental group when compared with the blank control group with time passing. The liver function test showed that alkaline phosphatase in the experimental group was significantly lower than that in the blank control group at 2 and 3 months (P lt; 0.05), but no significant difference in aspartate aminortransferase, total bilirubin, and albumin was found among 3 groups (P gt; 0.05). The histology and immunohistochemistry staining observations showed that the regeneration rates of submucosal glands and epithelium in the experimental group were faster than those in the blank control group; defect was covered with the epithelium at 2 months, and the structure was similar to that of normal control group at 3 months; and the edema and inflammation infiltration were reduced when compared with the blank control group. The counts of MVD and submucosal gland were significantly higher than those in blank control group and normal control group at 1 month after operation (P lt; 0.05), and then decreased and remained at normal levels at 2 months after operation. There was a positive correlation between submucosal gland counting and MVD counting in 3 groups after operation (P lt; 0.01). The cholangiography examination showed no biliary dilatation or cholelithiasis after 3 months in experimental group and blank control group. Conclusion Sustained-release bFGF can promote healing of bile duct defect by accelerating the vascularization, gland regeneration, and epithelialization.
Objective To observe the blood circulation compensation in the involved area of the liver following ligation of the third grade branches of hepatic artery and portal vein and bile duct enclosed in Glisson’s capsule. Methods Ligation of the third grade branches of these ducts was carried out in 7 pigs. Uptake of 99mTc-EHIDA in the liver was scanned with SPECT pre-and post-operatively. Liver angiography of hepatic artery and portal vein were taken at regular interval. Corrosion casts of these ducts were made with ABS following extirpation of the liver at the end of experiment. The histological specimens were examined with electronic microscope. Results Compensatory circulation occurred between involved and noninvolved part of the liver through the sinusoids in 30-60 minutes after ligation. In the 6 weeks following the procedure, there was also blood supply in the affected region of liver, and collateral developed through hepatic aterioles and capillaries. Conclusion Liver has an ability to establish compensatory blood supply on the condition of ischema in a local region of liver.
Objective To discuss the effective surgical treatment of intrahepatic lithiasis combined with high hepatic duct strictures. MethodsTwo hundreds and sixteen cases of intrahepatic lithiasis and high hepatic duct strictures treated in this hospital from January 1993 to October 2002 were analysed retrospectively.ResultsOne hundred and eightythree cases underwent different selective operation by selected time; 33 cases complicated with acute obstructive suppurative cholangitis underwent emergency were performed single biliary drainage, in which 30 cases were reoperated. The operative procedure were: hepatic lobectomy,high cholangiotomy and plastic repair,exposure of hepatic duct of the 2nd and the 3rd order,and plastic repair with own patch and choledochojejunostomy.Two hundreds and six cases were cured,the curative rate was 95.4%; 8 cases improved (3.7%), and 2 cases died (0.9%).Conclusion The best effective surgical treatment of intrahpatic lithiasis is hepatic lobectomy. Exposure of hepatic duct of the 2nd and the 3rd order is a satisfactory to release the hepatic duct strictures and to clear the intrahepatic lithiasis. For patients with normal extrahepatic bile duct and Oddi’s function, plastic repair of bile duct with own patch is possible to keep the normal form and function. Cholangioscopy may play an important role in the treatment of intrahepatic tract lithiasis during operation.