目的:研究离体肝脏缺血再灌注期间丝裂原活化蛋白激酶(mitogen activated protein kinase,p38MAPK)信号转导途径对肿瘤坏死因子α (tumor necrosis factor α,TNFα )mRNA表达的影响。方法:建立兔离体肝脏缺血再灌注模型,对照组(n=12)灌注液中不加特异性p38MAPK抑制剂SB202190,抑制组(n=12)灌注液中加入SB202190(浓度为3μmol/L)。分别于肝脏离体前,冷保存末,再灌注10min、30min、60min及120min时获取离体肝组织标本。应用Western-blot法及免疫沉淀法检测离体肝组织中p38MAPK表达的水平及活性,RT-PCR法检测TNF-α-mRNA表达水平。结果:对照组p38MAPK活性在冷保存末及再灌注10min、30min、60min均较离体前和再灌注120min显著升高(Plt;0.01),也显著高于同时相点的抑制组(Plt;0.01);抑制组p38MAPK活性在组内各时相点的变化无显著性差异(Pgt;0.05)。两组肝脏于离体前、冷保存末及再灌注10min及30min,肝组织中仅有少量TNF-α mRNA表达,组间及组内比较无显著性差异(Pgt;0.05);至再灌注60min及120min,对照组TNF-α mRNA的表达水平显著性高于组内其它时相点(Plt;0.01),而抑制组虽然也显著高于组内其它时相点(Plt;0.05),但却显著性低于同时相点对照组的表达水平(Plt;0.01)。离体再灌注期间供肝组织中p38MAPK活性与供肝组织内TNF-α mRNA的表达水平呈显著性正相关(r=0.996,Plt;0.01)。结论:p38MAPK对TNF-α生成的调节作用层次可能在转录水平,提示p38MAPK信号转导途径对TNF-α mRNA的调节可能是导致离体肝脏缺血再灌注损伤的重要机制之一。
目的:研究离体肝脏缺血再灌注期间丝裂原活化蛋白激酶(mitogen activated protein kinase,p38MAPK)信号转导途径对细胞间黏附分子1(intercellular adhesion molecular 1,ICAM1)mRNA表达的影响。方法:建立兔离体肝脏缺血再灌注模型,对照组(n=12):灌注液中不加特异性p38MAPK抑制剂SB202190,抑制组(n=12):灌注液中加入SB202190(浓度为3μmol/L)。于肝脏离体前,冷保存末,再灌注10min、30min、60min及120min时获取离体肝组织标本。分别应用Western-blot法及免疫沉淀法检测离体肝组织中p38MAPK表达的水平及活性,原位杂交法检测ICAM1 mRNA表达水平。结果:与离体前相较,对照组p38MAPK活性在冷保存末及再灌注10min、30min、60min显著性增高(Plt;0.01),而再灌注120min时活性与离体前相较无明显差异(Pgt;0.05);抑制组p38MAPK活性在各时相点的变化无显著性差异(Pgt;0.05),除离体前及再灌注120min两组肝脏的p38MAPK活性无显著性差异外,其余各时相点p38MAPK活性均显著性低于对照组(Plt;0.01)。离体前、冷保存末及再灌注10min及30min时,两组肝组织中仅有少量ICAM1 mRNA表达,组间及组内比较无显著性差异(Pgt;0.05);至再灌注60min及120min,对照组ICAM1 mRNA的表达水平显著性高于组内其它时相点(Plt;0.01),而抑制组虽然也显著高于组内其它时相点(Plt;0.05),但却显著性低于同时相点对照组的表达水平(Plt;0.01)。离体再灌注期间供肝组织中p38MAPK活性与供肝组织内ICAM1 mRNA的表达水平呈显著性正相关(r=0.985,Plt;0.01)。结论:p38MAPK对ICAM1生成的调节作用层次可能在转录水平,提示p38MAPK信号转导途径对ICAM1 mRNA的调节可能是导致离体肝脏缺血再灌注损伤的重要机制之一。
Objective To study the effect of p38MAPK activity on tumor necrosis factor-α （TNF-α） mRNA and intercellular adhesion molecule 1 （ICAM1） mRNA expressions of isolated rabbit liver during early stage of cold preservation and reperfusion period. Methods Based on the cold preservation and reperfusion model of isolated rabbit liver, the animals were divided into inhibition group （n=12） with 3 μmol/L SB202190 (p38MAPK specificity inhibitor) in perfusate and control group （n=12） with no SB202190 in perfusate. Liver tissue samples were harvested at the time points of before resection, end of cold preservation, and different reperfusion period (10, 30, 60 and 120 min). Protein expression and activity of p38MAPK were detected by Western blot and immunoprecipitation respectively, expression of TNF-α mRNA was detected by RT-PCR, and expression of ICAM1 mRNA was detected by in situ hybridization. Results There was no obvious change of expression of p38MAPK protein in liver tissue both in two groups during the total period （P＞0.05）， and there was no statistically significant difference between two groups （P＞0.05）. At time points of end of cold preservation, 10, 30 and 60 min of reperfusion, the activity of p38MAPK in control group was significantly higher than that at the time points of before resection and 120 min of reperfusion (P＜0.01), and was also significantly higher than that in inhibition group at the same time points (P＜0.01). There was no significant difference in activity of p38MAPK among all time points in inhibition group (P＞0.05). The expressions of TNF-α mRNA and ICAM1 mRNA at the time points of before resection, end of cold preservation, and 10 and 30 min of reperfusion were significantly lower than those in 60 and 120 min of reperfusion in both two groups (P＜0.05, P＜0.01); The expressions of TNF-α mRNA and ICAM1 mRNA in inhibition group were significantly lower than those in control group at the time points of 60 and 120 min of reperfusion (P＜0.01). The activity of p38MAPK of liver tissue during cold preservation and reperfusion period was significantly correlated with the level of TNF-α mRNA and level of ICAM1 mRNA expression （r=0.996, P＜0.01； r=0.985, P＜0.01）. Conclusions These results suggest that p38MAPK pathway may regulate the expressions of TNF-α and ICAM1 at the level of transcription and the activation of p38MAPK can up-regulate TNF-α and ICAM1 expressions, which may be one of the important mechanisms to cause ischemia-reperfusion injury of isolated liver during cold preservation and reperfusion period.
Objective To study the suitable operation method of elderly patients with acute cholecystitis. Methods The clinical data of 149 elderly patients with acute cholecystitis were retrospectively analyzed. All patients were divided into two groups according to the operation: open cholecystectomy group (OC group, n=76) and laparoscopic cholecystectomy group (LC group, n=73). Some clinical data were compared in this paper such as operation time, blood loss, length of hospital stay, time of resumption of food, time of intestinal function recovery and complications. Results No marked difference was found between OC group and LC group about basic data except WBC count and examination of gallbladder by B ultrasound（P＞0.05）. But there were significant difference in operation time, blood loss, time of resumption of food, time of intestinal function recovery, length of hospital stay and complications between OC group and LC group （P＜0.01）. Conclusion Individualized treatment should be emphasized on elderly patients with acute cholecystitis. Selection of OC or LC to these patients should be based on the clinical condition and taken the safety as the first principle.
Objective To expand the utilization of minimally invasive technologies for parapancreatic abscess, and summarize the application experience of choledochoscope for treatment of parapancreatic abscess. Methods The clinical data and treatment effectiveness of 36 patients with parapancreatic abscess from Dec. 2000 to Dec. 2008 were analyzed retrospectively. These patients had experienced percutaneous puncture and been placed drainage tube under the ultrasound guidance first, then expanded the sinus tract gradually, and performed debridement by choledochoscope. The flexibility of choledochoscope was used to remove the necrotic tissue and pyogenic membrane repeatedly by clamping, netting and vacuum aspiration in every domain. Results Thirty-six patients were performed percutaneous puncture and placed drainage tube, 3 cases were given canalis singularis, 7 cases were double tube, 26 cases were over three tube. The debridement times were 3-14 by choledochoscope, average 5.6 times. There were 6 cases with improving systemic symptoms, blood routine and temperature recovering normal, and drink and food recovering, then discharged from hospital with tube after 1-2 times of debridement. Length of stay was 25-132 d, average 76 d. The curing rate was 91.7% (33/36). Two cases were turned into open surgery because of broad necrotic tissue range combined with many abdominal cavity abscess with good postoperative recovery and cured. One case was dead of severe multiple organ failure combination. There were 2 patients with hemorrhage, 3 patients with external intestinal fistula. Conclusions The debridement of choledochoscope for parapancreatic abscess treatment is a simple, flexible and effective method. It changes the viewpoint that parapancreatic abscess can be cured only by operation drainage, decreases the patients’ trauma and accomplishes the idea of damage control by minimally invasive technologies.
Objective To detect the anti-colon cancer ability of whole cell lysates pulsed dendritic cell (DC) which acts as an adjuvant. Methods Whole cell lysates of LoVo cells were extracted with freeze thawing method, then the monocyte-derived DC were pulsed with this cellular antigen. Subsequently, the capability of antigen pulsed DC to promote T lymphocytes proliferation and the ability of T lymphocytes to kill LoVo cells were detected by 3H-TdR incorporation and lactate dehydrogenase release methods, respectively. Results The whole cell lysates of LoVo cells pulsed DC significantly stimulated T cells proliferation, and the cytotoxicity abilities of primed T cells to kill LoVo cells were also enhanced. Conclusion Tumor cell lysates which act as the cellular antigen to pulse DC can improve the efficacy of anti-cancer immune response， which provides us an experimental evidence for cancer immunotherapy.
ObjectiveTo compare the therapeutic efficacy of biliary tract stent placing for malignant obstruction of biliary tract by percutaneous transhepatic cholangiography (PTC) or endoscopic retrograde cholangiography (ERC). MethodsPTC approach: choosing the expansion intrahepatic bile duct which had a large angle traveling with common bile duct at the ultrasound-guided, we performed bile duct puncture and inserted the drainage pipe into it, then stent was placed with 68 cases (2 cases among the total were failure of ERC approach) after a week drainage. ERC approach: inserting drainage tube into the common bile duct by duodenal endoscopic retrogradely, the angiography showed obstruction site, the guide wire inserted through the obstruction site, then stent was placed along the guide wire with 53 cases. ResultsThe achievement ratio of stent placing by PTC was 100%(68/68), and which by ERC was 96.2%（51/53）. The complications (bleeding, bile leakage) didn’t happen in two groups. 1-18 months (average 12.4 months) of follow-up, the died cases of PTC group and ERC group were 7 and 5 cases within 6 months, respectively; the survive cases of which were 17 and 9 cases after 18 months of treatment, respectively. ConclusionsThe biliary tract stent placing is a safe and effective method to the malignant obstruction of biliary tract patients who can not drainage tube be treated by operation. It can relieve biliary obstruction efficiency, and can increase live time and life quality for patients. We can choose the stent placing method by ERC for cases whose obstruction site is at the inferior of common bile duct or duodenal ampulla, and the cases whose obstruction site is at the above of hepatic porta should be chosen by PTC.
Objective To summarize the experience of applying biliary balloon dilator to prevent rebleeding after operation for hepatolithiasis combined with hemobilia. Methods From 2003 to 2008, 11 patients were reoperated to stop from hemobilia by biliary balloon dilator’s application after operation for hepatolithiasis combined with hemobilia, and whose clinical data were collected and analyzed. Results In 11 cases, 7 were males and 4 were females. Eight cases were transferred from other hospitals. When intrahepatic duct bleeding was stopped, the biliary balloon dilator was placed at the right site under the guide of choledochoscope. Hemobilia occurred in 4 patients and biliary balloon dilator was opened to compress for 2 h, then decompress for 0.5 h alternately. In all of 4 patients, bleeding was stopped successfully, of which, 1 patient got hemobilia again 5 d after the first bleeding, and was also stopped by the same method. Conclusions After operation on hepatolithiasis combined hemobilia, rebleeding happens in some cases. Preset of biliary balloon dilator at the prebleeding site and opening it when rebleeding happens can get instant hemostasis, which may be a simple and effective treatment choice.