ObjectiveTo investigate the change of renal endothelin (ET) excretion and its relation to renal dysfunctions in obstructive jaundice.MethodsSixty male Wistar rats were randomized into two groups, the common bile ducts were ligated to establish the model of obstructive jaundice in experimental group, and only sham operation was done in control group. Ten rats were taken from each group at 5, 10 and 15 days respectively after operation, renal functions were evaluated by paminohippuric acid clearance (CPAH), inulin clearance (CIN) and fractional sodium excretion (FENa+); furthermore, plasma endotoxin (EX) level was determined, and ET1 contents in renal arterial plasma, renal venous plasma and renal tissue were detected. ResultsOnly FENa+ was significantly increased at the 5th day in experimental group; since the 10th day, all the three renal functional parameters gradually decreased, and FENa+ was significantly lower than that in control group at 15th day (P＜0.01 vs control). ②The plasma EX sustained at significantly higher levels after operation in experimental group (P＜0.01 vs control). ③The renal arterial plasma ET1 was significantly decreased, while the contents in renal venous plasma and renal tissue were significantly increased after operation in experimental group (P＜0.01 vs control). ④There were positive correlation between plasma EX and renal ET1 content, negative correlation between renal ET1 content and CPAH/CIN, and positive correlation between renal ET1 content and FENa+ (P＜0.01).ConclusionThe increased excretion of renal ET stimulated by endotoxemia may play an important role in the renal dysfunctions in obstructive jaundice.
Objective To investigate the regulatory effect of somatostatin analogue (SMS201995,SMS) on proliferation and apoptosis in human cholangiocarcinoma cell line in vitro. MethodsProliferation curve, flow cytometry, agarose gel electrophoresis, Annexin VFITC and flow cytometric immunofluorescent technique were performed to identify the inhibitory effect on cell proliferation and the induction of apoptosis of human cholangiocarcinoma cells (SKChA1). ResultsSMS significantly reduced the SKChA1 cell growth by serum in long experiments and transiently accumulated it in G0/G1 phase. Dotplot analysis of cells duallabeled with Annexin VFITC and PI confirmed the induction of apoptosis by SMS in SKChA1 cells.AnnexinVFITC labeling was markedly enhanced following treatment with SMS for 24 h. DNA of treated SKChA1 cells appeared a ladder pattern characteristic of apoptosis. Besides, timedependent increase in bax and decrease in bcl2 occured during SMS treatment. Conclusion SMS could inhibit the proliferation activity and induce apoptosis of cholangiocarcinoma cell line SKChA1. The mechanisms of apoptosis might be correlated with the expression of apoptosisregulatory gene bax and bcl2.
Objective To investigate the expression of multigenes mediated by adenovirus in liver cancer cells and the effects on growth of cells transducted with multigenes. MethodsBy construction of recombinant adenovirus containing human p53, B7-1, GM-CSF, and IL-2 genes (Ad-multigenes), the expression level of target genes in three human hepatocellular carcinoma cell lines and a human hepatocellular cell line L02 was detected using ELISA, immunohistochemistry and FACS assay and the change of growth of these cells and the tumor cell apoptosis were observed. Results The human hepatic cells and liver cancer cells were all sensitive to adenovirus infection. At a MOI of 50 PFU/cell, among the cells examined nearly 90% were positive expression and except IL-2, other three genes were expressed with high efficiency. The growth of Ad-multigenes-transduced liver cancer cell lines was inhibited and apoptosis was induced, but the growth of Ad-multigenes-transduced normal hepatic cell line L02 did not change. Conclusion These results indicate that the adenovirus is an efficient vector for gene transfer into human liver cancer cells. These liver cancer cell lines transduced with multigenes constructed on one recombinant adenoviral vector can highly express target genes and their growth was inhibited, and apoptosis appeared.
Objective To provide experimental evidence for the clinical application of ischemia therapy to treating pancreatic cancer. Methods After the model of pancreatic transplanted cancer was established in nude mice with orthotransplantation of human pancreatic cancer cell line into the pancreas, the ischemia of the right lobe of the pancreas was induced with ligation of the gastroduodenal, inferior pancreaticoduodenal and dorsal pancreatic arteries. Effects of regional ischemia on the growth of transplanted cancer and the pathomorphology of the transplanted cancer and pericancerous tissue were investigated. Results The transplanted cancer grew slower and its doubling time was longer in the ischemic group than in the control. On the 3rd, 7th and 14th day after operation, the size of transplanted cancer, the proliferative index and protein content of the cancer cells were significantly lower in the ischemic group than in the control (Ｐ＜0.01). Optical microscopy revealed large areas of coagulation necrosis, necrobiotic cells and the infiltration of inflammatory cells. The atrophy of acini, fibrosis and the infiltration of lymphocyte cells were found in pericancerous tissue. Conclusion Regional ischemia can destroy and inhibit the pancreatic transplanted cancer in nude mice effectively. The ischemia changes of pericancerous tissue may be unfavourable for the growth of the pancreatic transplanted cancer.
Objective To establish the model of hepatic VX2 tumor in rabbits and to offer the experimental evidences for the application of the model. Methods The hepatoma model was reproduced with VX2 cell lines in rabbits. The method to reproduce the model was improved. The changes of liver function (ALT, AST and TB) were determined at a different phase. Tumor’s growth and metastases, pathological changes, images and spontaneous survival time of the animal were observed. Results The tumors could grow up to 1.5-2.0 cm in diameter in 3 weeks after implanting. The successful rate of implantation was 100%. Nodular enhanced echo was found in the liver by color ultrasound. CT scans showed the low density foci in liver, while enhanced CT scans demonstrated asymmetrical intensification in the foci. Macroscopic observation showed that the tumors were grayish white in color and felt harder, necrotic foci was present in the center of tumor. Observation with light microscope showed that the tumor cells’ nucleoplasm proportion was great, tumor cells arranged irregularly, and the tumors displayed invasive growth and no obvious envelope around them. Animals’ spontaneous survival time was 40-53 days. The cause for their death was multiple system organ failure. Conclusion In pathological morphology, pathological process and prognosis, the hepaticVX2 tumors in rabbits are similar to human hepatocarcinoma. It has such characteristics as easy reproduction, short growth period, high success rate, high stability and so on. The model is an ideal hepatoma model in animals.