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.
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】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 investigate the effects and mechanisms of bile on small intestine mucosal barrier.Methods Fifty Wistar rats were assinged into 3 groups randomly: obstructive jaundice (OJ) group （n=20）, biliary external drainage group （n=20） and control group （n=10）. Ten days after operation, the plasma endotoxin level was determinated, the terminal ileum mucosas was obtained to be morphologically measured by light microscope, and immunohistochemistry and Western blot were uesd to examine the expressions of tight junction proteins zona occludens-1 (ZO-1) and occludin in the mucosas. Results Atrophy significantly appeared in the distal ileum mucosas in OJ group. Compared with control group, the intestinal villus height, mucosa thickness and crypt depth in OJ group were obviously decreased 27.8%, 21.7%, and 25.4% (P=0.001, 0.001, 0.040). There were no differences between external drainage group and control group (P=0.050, 0.070, 0.080); While the values of external drainage group were significantly higher than those in OJ group (all P=0.001). The level of plasma endotoxin was up to (1.49±0.27) EU/ml in OJ group compared with control group 〔（0.27±0.09） EU/ml〕, P=0.001. In external drainage group, the value was （0.91±0.25） EU/ml, which was obviously higher than that in control group and lower than that in OJ group （all P=0.001）. Immunohistochemical study showed b positive expression of ZO-1 dropped from 7/10 in the control group to 6/20 in OJ group （P=0.040）, occludin expression was 8/10 in control group and 7/20 in OJ Group （P=0.020）; expressions of them in external drainage group 〔8/20 （P=0.100,0.210） and 9/20 （P=0.060, 0.200）〕 displayed no significant differences compared with the other twogroups. Quantitative testing of Western blot showed the expressions of ZO-1 and occludin in OJ group were significantly lower than those in control group (P=0.001, 0.010), the values in external drainage group were higher than those in OJ group （P=0.005, 0.014）. The expression of ZO-1 was lower in external drainage group than that in control group (P=0.001), and there was no significant difference of occludin between the two groups (P=0.062）. Conclusion Lack of intestinal bile will undermine the intestinal tight junction protein composition, and make intestinal mucosal barrier impaired. The intestinal barrier more severely injured when biliary tract obstructs because of multiple factors. Bile plays an important role in the maintenance of intestinal mucosal barrier.
【Abstract】ObjectiveTo explore risk factors of bile leakage after primary ductal closure following choledochotomy. MethodsA retrospective clinical analysis was made in 148 cases of Ttube drainage and 154 cases of primary common bile duct suture following choledochotomy admitted to our hospital from January 1999 to June 2003. Results Postoperative bile leakage was seen in 11 patients of the group with Ttube drainage and in 16 patients of the group with primary suture respectively, there was no significant difference(Pgt;0.05).In the group with primary common bile duct suture,the occurrence of bile leakage was relative with hyperglycemia(Plt;0.05)，hypoproteinemia (Plt;0.01)，bile duct repeated sutures(Plt;0.01)and positive bacterial culture in bile(Plt;0.05). Bile leakage was not relative with elder age (Pgt;0.05).Conclusion It is the key factors,including chosing appropriate patients, intraoperative special examination, careful manipulation and effective medical treatment that can reduce the morbidity of bile leakage.
Objective To study the earlier and later clinic results of 64 chronic duodenal ulcer patients treated with high selective vagotomy and mucosal antrectomy (HSV+MA). Methods The clinic results of the patients and the changes of gastrin, motilin and somatostatin in the blood were prospectively investigated. Results Fifty nine (92.2%) patients after 3-6 months of follow-up and 26 (92.9%) patients after 5-8 years of follow-up achieved Visick grates Ⅰ-Ⅱ. No patients died. Gastric acid secretion and infection rate of helicobacter pylori in antral mucosa were significantly reduced after operation. No significant difference was showed in bile acids and total bacterial counts of gastric juice before and after operation. No ulcer recurrence was found by barium meal and endoscopy. There was no significant difference in serum gastrin and plasma motilin before and after operation. The level of somatostatin in the blood of patients after 5-8 years of follow-up was decreased. Conclusion HSV+MA is the better operative treatment for duodenal ulcer, since it can not only effectively and lastingly decrease acid secretion and rates of ulcer recurrence, but also preserve the function of the antrum and pylorus and keep the gastric milieu interne relatively stable.
ObjectiveTo evaluate the clinical manifestation, diagnosis, and treatment of intrahepatic biliary cystadenoma (IBCA). MethodsThe clinical date of 12 patients with IBCA from January 2004 to December 2013 in our hospital were retrospectively analyzed. ResultsOf the total 10 patients were female and 2 were male. The average age was 49.0 years old (16-77 49.0 years old). The course of disease were 1 month-3years (average:7 months). The clinical symptoms were right upper abdominal fullness with a sense of pain in 6 patients, right upper abdominal mass in 1 case, jaundice in 1 case, and no symptoms in 4 cases. Enhanced CT examination showed that the tumor complete capsule and there were internal septations in 11 cases, the density of internal septations was different. There was mild enhancement of cyst wall and internal septations in 9 cases. All of 12 cases received surgical resection. Only 1 case showed recurrence in 2 years after operation, the remaining 11 patients had no recurrence. ConclusionsIBCA often occurs in middle-aged women. The main clinical symptoms are abdominal fullness with a sense of pain, right upper abdominal mass or jaundice. Enhanced CT is the main method of diagnosis before operation. Radical resection of IBCA is the best treatment, and can prevent recurrence effectively.
To study the prognostic significance of proliferative cell nuclear antigen(PCNA)in bile duct carcinoma,expression of PCNA protein was studied immunohistochemically in 30 patients with bile duct carcinoma.Results:86.7 percent of bile duct carcinomas showed PCNA positive staining and significiant difference was observed between malignant and benign tissues.These results suggest that proliferative activity of malignant tissues was ber than that of the benign ones.In patients with cancer of stages 3,the mean survial time for those with high proliferative activity was 13 months in constrast with 26 months for those with low activity.PCNA is of prognostic significance for bile duct carcinoma patients.
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.
In order to study rheologic property of bile flow between gallbladder and biliary duct during biliary obstruction,we made a model of complete biliary obstruction(CBO)in dogs.The results showed that:①The behavior of bile flow between gallbladder and biliary duct in noemal dogs belonged to Casson flow;②When the duration of CBO prolonged,the behavior of bile flow between gallbladder and biliary tract in the CBO dogs still belonged to Casson flow.The changes of yield stress and apparent viscosity at high or low shear rate in bile flow of the biliary duct were similar to that in bile flow of the gallbladder.