Objective To investigate the prevention and treatment strategy of bile duct injury (BDI) in laparoscopic cholecystectomy (LC). MethodsLatest progress was reviewed based on recent documents and the experience on BDI in LC in our department. ResultsWith the popularity of LC, BDI in LC is increasing. The reasons include illegibility and variability of local anatomy in gallbladder trigone,injury caused by galvanothermy, as well as operator’s overconfidence. In order to prevent BDI, we should apply more blunt dissection, not to use electrocogulation if possible and to study local anatomy and its variance clearly. The common bile duct and common hepatic duct should be clearly identified. Intraoperative cholangiography, laparoscopic ultrasonography and hepatobiliary scintigraphy are selections as necessary.The treatment of BDI depends on the type of BDI and its site and local condition.The treatment includes end to end anastomosis, repairing the defect, choledochoduodenostomy, RouxenY choledochojejunostomy and so on. T tube should be maintained in place for more than half a year after operation.Conclusion The key to improve the prognosis of BDI is prevention and treatment in proper time and in correct way.
【Abstract】Objective To study the regulatory ability of peroxisome proliferatoractivated receptor γ(PPARγ) ligands to the inflammatory response in human gallbladder epithelial cells. Methods Culture human gallbladder epithelial cells and identify them . Cells were treated for 24 hours with 0, 10 μmol/L, 20 μmol/L, 30 μmol/L, 50 μmol/L and 100 μmol/L of Ciglitazone during cellular growth peak(5th day), then stimulated them with hIL-1β 5 ng/ml for 2 hours and measured the concentration of IL-6、IL-8 and TNF-α in cellular supernatants by riadioimmunoassay. Results Contrasted with control group, the expression of IL-6 and IL-8 in each test group were inhibited （P＜0.001）. The IL-6 and IL-8 levels were gradually dropped and corelated with the dosage of Cigtitazone, and manifested dosagedependence (P＜0.001). The concentration of TNF-α could not be measured. Conclusion PPARγ ligands can inhibit the expression of IL-6 and IL-8 in human gallbladder epithelial cells and probably produce effect in the regulation of cholecystic inflammation.
【Abstract】ObjectiveTo construct a mrp1 expression vector and investigate its biological characteristics in HepG2 cells in vitro. MethodsThe 6.5 kb multidrug resistanceassociated protein （MRP) cDNA obtained from plasmid pGEM-mrp1 was cloned into the pCI-neo mammalian expression vector, which was later transferred into human hepatocarcinoma cell line HepG2 by liposome. Then the HepG2 cells resisting G418 were clustered and proliferated, and the mrp1 mRNA and MRP in these HepG2 cells were detected by means of RT-PCR and FCM respectively. ResultsThe mrp1 expression vector was established successfully, and the stable MDR hepatocarcinoma cell line (HepG2/mrp1) was developed as well. The content of the specific fragment of mrp1 mRNA was (56.8±6.37)% and MRP was 7.89 in the HepG2/mrp1 cells, the corresponding value in HepG2 cells was (9.67±3.26)% and 0.79 respectively. The difference was statistically significant (P＜0.05). ConclusionIt is practicable to establish MDR hepatocarcinoma cell line by transferring mrp1 cDNA into HepG2 cells, which is useful in the research of MDR mechanism.
ObjectiveTo explore the suitable method for isolation and maintenance of primary cultures of human gallbladder epithelial cells (GECs) for establishing the basis of research works in physiological function of gallbladder and its related diseases.MethodsGECs were isolated with collagenase type Ⅳ and blunt separation.The dishes were coated with fibronectin, laminin and polyDlysine respectively.Additional 10 ng/ml epidermal growth factor was added to DMEM medium containing 20% fetal calf serum.The cells were studied under light and electron microscope to determine their shape and distribution.ResultsEach gallbladder yielded approximately (1-5)×107columnar epithelial cells,greater than 95% of which were viable by trypan blue exclusion.The cells grew vigorously within one week which was flat and multangular in shape. CK19 expressed positive.Electron microscope showed typical gallbladder epithelia with microvilli,tight junctions and mucus droplets.ConclusionCombination of collagenase type Ⅳ，mechanical blunt separation and twostep attachment is of great benefit for separating and harvesting GEC.Fibronectin coated culture dish and DMEM medium containing 20% calf serum and 10 ng/ml hEGF is of great benefit for culturing gallbladder epithelial cells.
ObjectiveTo evaluate the operative technique and clinical efficacy of laparoscopic splenectomy (LS) combined with esophagogastric devascularization in treatment of portal hypertension induced by liver cirrhosis. MethodsTwelve cases with esophageal and gastric varices induced by portal hypertension and liver cirrhosis were treated by the LS combined with esophagogastric devascularization in our department from March 2009 to August 2010, which clinical data were analyzed and summarized retrospectively. ResultsThe splenic artery was ligated before the treatment of splenic pedicle in 12 cases, LS combined with pericardial devascularization was successfully performed in 10 cases, 7 cases of which were treated by the level two transection method of splenic pedicle, and 2 cases were converted to open surgery due to intraoperative bleeding. In 10 cases, the operative time was 180-300 min (average 210 min), and intraoperative blood loss was 200-1 000 ml (average 480 ml). The postoperative hospital stay was 8-15 d (average 9 d)， the postoperative complications included plural effusion （lt;300 ml） in 2 cases， mild ascites （lt;300 ml） in 2 cases, and mild pancreatic leakage in 1 case， but all were cured eventually， and no mortality occurred. Followup was conducted in 12 patients for 4 to 20 months (average 7 months), and no rebleeding occurred. ConclusionsLS combined with pericardial devascularization is relatively safe and effective methods in treatment of portal hypertension induced by liver cirrhosis. The keys to success include ligation of splenic artery, and the use of harmonic scalpel combined with ligasure to treat splenic pedicle.