ObjectiveTo study the relationship between hepatocellular apoptosis and glycogen contents during hepatic cold preservationreperfusion and its mechanism.MethodsBased on the model of four groups of rabbit livers with different hepatocellular glycogen contents, hepatocellular apoptosis and bax gene expression were observed during hepatic cold preservationreperfusion.ResultsApoptotic hepatocytes were obviously found in 60 minute reperfusing livers subsequent to 9 hour cold storage, and there was significant difference in the numbers of apoptotic hepatocytes among all the groups. In the same time, there was the close relationship between the levels of bax gene expression and the glycogen contents of hepatocytes.ConclusionIntracellular abundant glycogen may significantly depress the hepatocellular apoptosis during hepatic cold preservationreperfusion by decreasing hepatocellular bax gene expression.
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 find the relation between donor liver cold preservation-reperfusion injury and hepatocellular apoptosis during liver transplantation. Methods Four groups of rabbit livers, which had experienced cold storage for 0,3,6,9hr respectively, were observed during their cold preservation-reperfusion by using TdT-mediated dUTP-biotion nickend labeling and electron microscope.Results Apoptopic hepatic parenchymal cells were obviously observed in reperfusing livers subsequent to cold storage. Furthermore, the longer the cold storage duration, the greater the number of apoptotic cells. On the contrary, no or rare apoptotic hepatic parenchymal cells was observed in all the groups at the end of cold preservation. Conclusion It suggests that apoptosis of hepatic parenchymal cells is markedly involved in donor liver cold preservation-reperfusion injury.
目的考察皮下通道型胆囊肝胆管成形术（STHG）治疗肝胆管结石及胆管狭窄的中、远期疗效。方法对该院1994年12月至2000年6月期间行STHG手术的59例患者的术后中、远期并发症进行统计分析。结果STHG的术后并发症发生率较低，而且并发症的种类也较少； 本组病例术后无返流性胆管炎的表现，也无胃肠道功能紊乱和吻合口溃疡发生。结论STHG既保存了胆囊及Oddi括约肌功能，又保证了胆汁的生理流向，还能防止肠液的返流，从而避免了术后消化功能紊乱和返流性胆管炎的发生，是一种较为理想的治疗肝胆管结石和肝门部胆管狭窄的术式。