The Clinical And Experimental Study Of Anomalous Pancreaticobiliary Ductal Union

Anomalous pancreaticobiliary ductal union is a congenital anomaly defined as common bile duct and pancreatic duct union that is located outside the duodenal wall and beyond the influence of the sphincter of Oddi. Normally, the common bile duct and pancreatic duct open into the duodenum via a common channel which is usually less 10 mm and it is considered anomalous when this is more than 15 mm in adults and 5 mm in children. What the sphincter of Oddi falls to prevent mutual reflux of bile and pancreatic juice into the pancreatic and bile ducts lead to some diseases such as congenital choledochal dilation, cholangitis and carcinoma of the gallbladder so on. Anomalous pancreaticobiliary ductal union had been studied fully in clinic since it was reported by Kozumi and Kodama in 1916.However the etiology and pathophysiology are still unknowen and most of researches are on the retrospective of the clinical features and imageing findings. In order to obtain more information, we investigate the pancreaticobiliary dissection in foetus to know the shapes and the locations of the greater duodenal papilla, the types of pancreaticobiliary ductal union. We also produce an animal model for anomalous pancreaticobiliary ductal union to study the injuries of livers, pancreas and gallbladders in cats with anomalous pancreaticobiliary ductal union. Besides, the anti-oxidative treatment in cats with anomalous pancreaticobiliary ductal union were evaluated here.1.The study of pancreaticobiliary dissection in foetusObjective: The pancreaticobiliary dissection in foetus were investigated to know the shapes and the locations of the greater duodenal papilla, the types of pancreaticobiliary ductal union. Methods: 36 foetus who had died of induction of labor with water bag in 6 hours about 4 to 9 months old were studied. The specimens, including gallbladder, common bile duct, pancreatic duct and duodenum, were preserved in 10% buffered formaldehyde solution for 24 hours. Then the locations and shapes of the greater duodenal papilla were observed. Sections measuring 6μm were cut continuously from the paraffin-embedded tissue block, one was selected in every five sections and stained with hematoxylin and eosin. Then the sections were observed under the microscope.Results: 1) The shapes of the greater duodenal papilla showed three kinds of patterns including hemispheroid(58.1%) , circular cylinder(25%) and applanation(16.9%).The greater duodenal papillas locataed at the upper pars descendens duodenum (8.3%),the middle pars descendens duodenum(69.4%),the lower pars descendens duodenum(19.4%) and the farer(2.9%).2)The types of pancreaticobiliary ductal union were classified into four types , known as"Y"in 24 specimens(66.7%),"U"in 4 specimens(11.1%) ,"V"in 7 specimens(19.4%) and APBDU type in 1 specimens(2.8%).Conclusions: There are many kinds of types in pancreaticobiliary ductal union which has great clinical significance.2.Expression of proliferating cell nuclear antigen in epithelial cells of bile ductObjective: To examine the proliferation and differentiation of epithelial cells in bile duct by checking their expression of proliferating cell nuclear antigen through the cases of congenital biliary dilatation, biliary cancer and normal bile duct.Methods: 20 cases of congenital biliary dilatation, 8 cases of biliary cancer and 20 cases of normal bile duct were studied in the experiment. The specimens were preserved in 10%buffered formaldehyde solution for 24 hours, embedded in paraffin, slided 3μm thick for proliferating cell nuclear antigen histochemical studies. Results: Proliferating cell nuclear antigen labeling was 5.45%±1.90% in 20 cases of normal bile duct,47.75%±5.48% in 20 cases of congenital biliary dilatation and 83.88%±7.24% in 8 cases of biliary cancer. The expression of proliferating cell nuclear antigen showed a highly difference among them.Conclusions: There are difference expression of proliferating cell nuclear antigen in these three kinds of epithelial cells of bile duct. Proliferating cell nuclear antigen may be a better indicater to evaluate the proliferation and differentiation of epithelial cells in bile duct3.An animal model for anomalous pancreaticobiliary ductal union.Objective: To produce an animal model for anomalous pancreaticobiliary ductal union.Methods: 15 healthy cats, fasted about 12 hours before operation, anaesthetized by 3.5% pentobarbital sodium(1.3ml/kg body-weight) were performed in the experiment, whose abdominal wall were opened by a vertical incision of approximately 6 cm. The capsule of pancreas was opened to expose the pancreatic duct near which the common bile duct enters the duodenum. A 4- to 6-mm incision was made in the pancreatic duct and the common bile duct. The incised edges were anastomosed with continuous 6-0 nylon monofilament suture to make a wide and long communication between the pancreatic duct and the common bile duct, resembling the common channel of anomalous pancreaticobiliary ductal union in humans. The cholangiography was made in 20 days later of operation.Results: After being operated, 11of cats had good spirit and appetite without showing dispirited and fantod. The cholangiography showed that there were a wide and long communication between the pancreatic duct and the common bile duct.4 cats died at the day of the operation. Conclusions: This animal model for anomalous pancreaticobiliary ductal union is better than the other model.4.The experimental study on the injuries of livers, pancreas and gallbladders in cats with anomalous pancreaticobiliary ductal unionObjective: To study the injuries of livers, pancreas and gallbladders in cats with anomalous pancreaticobiliary ductal union.Methods: 10 healthy cats which were made a wide and long communication between the pancreatic duct and the common bile duct, resembling the common channel of anomalous pancreaticobiliary ductal union in humans were performed in the experiment. After 6 months, the livers, pancreas and gallbladders of these animals were removed and histopathological changes were evaluated by pathological and electron microscopic examination. Amylase in bile ,PCNA and the level of MDA were tested.Results: Seven out of 10 cats survived surgery and remained alive for 6 months with significant changes observed.1)The changes in livers. Before operation, the color of liver was salmon pink and there were no nodus in liver. The normal hepatic cells without steatosis were observed under a light microscope. However, the color became deeper and a little nodus were founded in liver 6 months later. Histopathology showed ballooning changes and inflammatory infiltrated in liver cells. The scores of histopathology were higher than before operation (9:1) with marked differences(P<0.05). Mitochondria swelling and lipid droplet in intracytoplasm were observed under an electron microscope.2) The changes in pancreas. Before operation, the color of pancreas was salmon pink and the pancreatic duct was not found in the face of pancreas. The normal pancreatic cells without inflammation were observed under a light microscope. 6 months later, the pancreas became deeper in color with hyperaemia and hydroncus, which three of them could found the dilated pancreatic duct in the face of pancreas. Histopathology showed that three of cases were mixed inflammatory infiltrate with numerous neutrophils, vascular proliferation in four cases.3)The changes in biliary tract. 6 months later, the wall of gallbladder became thicker, the bile became thicker. The amylase concentration in the gallbladder bile 6 months later was significantly higher than that before operation(192.08±62.04IU vs 9368.09±2204.42IU.P<0.05). The gallbladder epithelium became villous in appearance 6 months later, although the epithelium of that before operation was flat with few folds. The epithelial cells of gallbladder arranged tightly, although 6 months later a significant change such as endoplasmic reticulum expanding, intercellular space broadening and cellular nucleus deforming. The positive cells of proliferating cell nuclear antigen were increased significantly in the gallbladder epithelium at 6 months later(7.29%±2.70% vs 54.71%±10.90%.P<0.05).Conclusions: This animal model demonstrated that anomalous pancreaticobiliary ductal union would be harmful to the liver, pancreas and gallbladder.5. The experimental study on the anti-oxidative to the injuries of anomalous pancreaticobiliary ductal unionObjective: To evaluate the anti-oxidative treatment in cats with anomalous pancreaticobiliary ductal union.Methods: 13 healthy cats which had been made a wide and long communication between the pancreatic duct and the common bile duct, resembling the common channel of anomalous pancreaticobiliary ductal union in humans were divided randomly into two groups: 7 cats in group A were not treated. 6 cats in group B were treated with melatonin injected intraperitoneally (1mg/kg/d) three months, and breeded with vitamin C(500mg/kg/d) and vitamin E(500mg/kg/d) in next three months. After 6 months, the livers, pancreas and gallbladders of these animals were removed and histopathological changes were evaluated by pathological and electron microscopic examination. The level of MDA were testedResults: 1)The changes in livers. 13 cats in control group showed that the color of live was salmon pink and no nodus in liver. In A group, the color of liver became deeper and a little nodus was founded in liver. Histopathology showed ballooning changes in liver cells, mitochondria swelling and lipid droplet in intracytoplasm.1 of cats in group B had the same changes as the group A, The other 5 cats had less changes than that in group A both in pathological and electron microscopic examination.2) The changes in pancreas. In control group, the color of pancreas was salmon pink and the pancreatic duct was not found in the face of pancreas.In group A, the pancreas became deeper in color with hyperaemia and hydroncus, which three of them could found the dilated pancreatic duct in the face of pancreas. In group B, no dilated pancreatic duct and hydroncus were found. Histopathology showed that three of cases were mixed inflammatory infiltrate with numerous neutrophils, vascular proliferation in four cases in group A. but no changes were found in group B.3)The changes in biliary tract. The wall of gallbladder became thicker and the bile became thicker in group A than that in control group. There were no differences between the group of A and B. The amylase concentration in the gallbladder bile was 9368.09±2204.42 IU in group A and 8746.25±2077.95 IU in group B. The gallbladder epithelium became villous in appearance with in group A, although the epithelium of that in group of control was flat with few folds. The epithelial cells of gallbladder arranged tightly in control group, while a significant change such as endoplasmic reticulum expanding, intercellular space broadening and cellular nucleus deforming were observed in group A. The things in group B were better in group A. The positive cells of proliferating cell nuclear antigen were 7.29%±2.70% in control,54.71%±10.90 % in group A, 48.17%±13.06% in group B. The levels of MDA were 1.095±0.653nmol/mgprot in control,2.745±1.533 nmol/mgprot in group A ,1.302±0.771nmol/mgprot in group B. Conclusions: The anti-oxidative treatment was effective to the anomalous pancreaticobiliary ductal unionl.