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 evaluate the relationship between endothelin (ET) in bile and peripheral blood with systemic and hepatobiliary injury in patients with acute cholangitis of severe type (ACST). Methods ET, ALT and total bilirubin in bile and peripheral veinous blood of 25 patients with acute cholangitis of severe type (ACST) were detected during operation, one week and two weeks after operation. Results The contents of ET, ALT and total bilirubin were significantly lower on 7-day and 14-day after operations as compared with that during operations (Ｐ＜0.05 and Ｐ＜0.01). The concentration of ET in peripheral veinous blood paralleled with that in bile. Conclusion This suggests that ET is tightly related with the pathologic process of ACST. So, in patients with ACST, the dynamic measurement of ET in peripheral veinous blood can be an index for judging the degree of pathological damage either to the hepatobiliary or systemic systems.
ObjectiveTo analyze clinical experience and outcomes of bileaflet preservation in mitral valve replace-ment (MVR) for patients with severe mitral regurgitation (MR). MethodsWe retrospectively analyzed clinical data of 17 patients with severe MR who underwent MVR with bileaflet preservation in the Department of Cardiovascular surgery of Guangdong General Hospital from June 2011 to January 2013. There were 14 males and 3 females with mean age of 63.41±11.82 years (range, 38 to 82 years). There were 13 patients with atrial fibrillation. Preoperatively, 5 patients were in New York Heart Association (NYHA) functional class Ⅲ, and 12 patients were in NYHA class Ⅳ. There were 7 patients with ischemic MR, 9 patients with degenerative MR, and 1 patient with rheumatic MR. ResultsMVR with bileaflet preservation was performed for all the patients. Concomitant coronary artery bypass grafting was performed for 4 patients. Eleven patients received bioprosthetic MVR, and 6 patients received mechanical MVR. There was no in-hospital death, postoperative low cardiac output syndrome or left ventricular rupture. All the 17 patients were followed up for a mean duration of 16.44±5.02 months (range, 2 to 25 months). During follow-up, 1 patient died of severe paravalvular leak 2 months after surgery. All the other patients had good mitral valve function. None of the patients had anticoagulation or prosthetic valve related complication. Patient's heart function was significantly improved. Eleven patients were in NYHA functional class Ⅰ, 4 patients were in NYHA class Ⅱ, and 1 patient was in NYHA class Ⅲ. Cardiothoracic ratio, left atrial dimension, left ventricular end-diastolic dimension and left ventricular end-systole dimension postoperatively and during follow-up were significantly smaller than preoperative values. Postoperative left ventricular ejection fraction (LVEF) was significantly lower than preoperative LVEF(50.94%±8.78% vs. 55.31%±10.44%, P=0.04), but LVEF during follow-up was not statistically different from preoperative LVEF(55.31%±10.44% vs. 56.13%±9.67%, P=0.73), and LVEF during follow-up was significantly higher than postoperative LVEF(56.13%±9.67% vs. 50.94%±8.78%, P=0.02). There was no statistical difference between postoperative mitral pressure half-time (PHT)and PHT during follow-up (95.06±19.00 ms vs. 94.56±19.19 ms, P=0.91). ConclusionMVR with bileaflet preservation is a safe and effective surgical technique for patients with severe MR, and can significantly improve postoperative left ventricular remodeling and function.
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.
摘要:目的: 探讨拔T管后出现胆漏的预防和治疗。 方法 :对1986～2006年间取T管后发生胆漏的12例临床资料作回顾性分析。 结果 :12例经保守治疗后均痊愈。 结论 :拔T管后胆漏的发生是多因素的结果,改进手术技巧,适当延长拔T管的时间有助于预防胆漏的发生。胆漏发生后及时用导尿管置入窦道作引流是首选的治疗方法。Abstract: Objective: To investigate the prophylaxis and treatment of biliary fistula after removal of Ttube. Methods : The clinical data of 12 patients with biliary fistula after removal of Ttube from 1986 to 2006 were analyzed retrospectively. Results : Fistula was cured with conservative treatment in 12 patients. Conclusion : Many factors may affect the development of biliary fistula after removeal of Ttube. Improvement of surgercial technique and the proper prolongation for the time of removal of Ttube may contribute to prophylaxis the biliary fistula. Inserting a Nelaton’s catheter into the sinus tract to drain is the first choice to treat the local bile peritonitis that has occurred.
【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.
【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 investigate whether intraductal electrocautery incision （IEI） could decrease the recurrence of post-liver transplant anastomotic strictures （PTAS） after conventional endoscopic intervention of balloon dilatation （BD） and plastic stenting （PS）. Methods The clinical data of 27 patients with PTAS who were given endoscopic treatment of BD＋PS or IEI＋BD＋PS in our hospital from January 2007 to October 2011 were reviewed retrospectively. Results The treatment of BD＋PS was initially successful in 9 of 11 （81.8%） cases， but showed recurrence in 5 of 9 （55.6%）. The treatment of IEI＋BD＋PS was initially successful in 14 of 16 （87.5%） cases, and the recurrence was observed only in 3 of 14 （21.4%）. The total diameter of inserted plastic stents in IEI＋BD＋PS group was significantly greater than that in BD＋PS group 〔（12±3.2） Fr vs. （8±1.3） Fr，P=0.039〕. All recurrences were successfully retreated by IEI＋BD＋PS. Procedure-related complications included pancreatitis in 5 cases （18.5%）, cholangitis in 8 cases （29.6%）, bleeding after EST in 1 cases （3.7%）, which were all cured with medical treatment. No complications related to intraductal endocautery incision procedure such as bleeding and perforation were observed. Median follow-up after completion of endoscopic therapy was 22 months （range 1-49 months）. Conclusions Intraductal electrocautery incision is an effective and safe supplement to balloon dilatation and plastic stenting treatment of PTAS, which can decrease the recurrence of anastomotic strictures in conventional endoscopic intervention.
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.
The quantitative studies of secretory granules andand lysosomes in gallbladder epithelium and gallbladder bile glycoprotein were performed in 20 gallstone patients and 15 gallstone-free subjects. The results showed that the number, the volume density and the total secrectory granules were significantly increased in gallstone patients compared with gallstone-free controls. The gallstone patients had a markedly reduced number,volume destiny and total lysosome area compared with gallstone-free subjects. The glycoprotein concenrtation in gallblader bile was increased up to 21.04±4.92g/L in gallstone paients,as compare with 13.02±5.72g/L(Plt;0.05)in the stone-free controls. The qualitity of secretory granules and lysosomes was directly proprtational to that of bile glycproteins. Evidence and secrectory granules and lysosomes was directly proporional to taht of bile glycoproteins. Evidence and argument are presentded suggesting that gallblader epithelium secrectory granules affect the concentration of bile glycoprotein and lysosomemay be related to the intracellular degradation of secretory granules.