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.
目的:研究離體肝臟缺血再灌注期間絲裂原活化蛋白激酶(mitogen activated protein kinase,p38MAPK)信號轉導途徑對細胞間黏附分子1(intercellular adhesion molecular 1,ICAM1)mRNA表達的影響。方法:建立兔離體肝臟缺血再灌注模型,對照組(n=12):灌注液中不加特異性p38MAPK抑制劑SB202190,抑制組(n=12):灌注液中加入SB202190(濃度為3μmol/L)。于肝臟離體前,冷保存末,再灌注10min、30min、60min及120min時獲取離體肝組織標本。分別應用Western-blot法及免疫沉淀法檢測離體肝組織中p38MAPK表達的水平及活性,原位雜交法檢測ICAM1 mRNA表達水平。結果:與離體前相較,對照組p38MAPK活性在冷保存末及再灌注10min、30min、60min顯著性增高(Plt;0.01),而再灌注120min時活性與離體前相較無明顯差異(Pgt;0.05);抑制組p38MAPK活性在各時相點的變化無顯著性差異(Pgt;0.05),除離體前及再灌注120min兩組肝臟的p38MAPK活性無顯著性差異外,其余各時相點p38MAPK活性均顯著性低于對照組(Plt;0.01)。離體前、冷保存末及再灌注10min及30min時,兩組肝組織中僅有少量ICAM1 mRNA表達,組間及組內比較無顯著性差異(Pgt;0.05);至再灌注60min及120min,對照組ICAM1 mRNA的表達水平顯著性高于組內其它時相點(Plt;0.01),而抑制組雖然也顯著高于組內其它時相點(Plt;0.05),但卻顯著性低于同時相點對照組的表達水平(Plt;0.01)。離體再灌注期間供肝組織中p38MAPK活性與供肝組織內ICAM1 mRNA的表達水平呈顯著性正相關(r=0.985,Plt;0.01)。結論:p38MAPK對ICAM1生成的調節作用層次可能在轉錄水平,提示p38MAPK信號轉導途徑對ICAM1 mRNA的調節可能是導致離體肝臟缺血再灌注損傷的重要機制之一。
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.
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.
目的:研究離體肝臟缺血再灌注期間絲裂原活化蛋白激酶(mitogen activated protein kinase,p38MAPK)信號轉導途徑對腫瘤壞死因子α (tumor necrosis factor α,TNFα )mRNA表達的影響。方法:建立兔離體肝臟缺血再灌注模型,對照組(n=12)灌注液中不加特異性p38MAPK抑制劑SB202190,抑制組(n=12)灌注液中加入SB202190(濃度為3μmol/L)。分別于肝臟離體前,冷保存末,再灌注10min、30min、60min及120min時獲取離體肝組織標本。應用Western-blot法及免疫沉淀法檢測離體肝組織中p38MAPK表達的水平及活性,RT-PCR法檢測TNF-α-mRNA表達水平。結果:對照組p38MAPK活性在冷保存末及再灌注10min、30min、60min均較離體前和再灌注120min顯著升高(Plt;0.01),也顯著高于同時相點的抑制組(Plt;0.01);抑制組p38MAPK活性在組內各時相點的變化無顯著性差異(Pgt;0.05)。兩組肝臟于離體前、冷保存末及再灌注10min及30min,肝組織中僅有少量TNF-α mRNA表達,組間及組內比較無顯著性差異(Pgt;0.05);至再灌注60min及120min,對照組TNF-α mRNA的表達水平顯著性高于組內其它時相點(Plt;0.01),而抑制組雖然也顯著高于組內其它時相點(Plt;0.05),但卻顯著性低于同時相點對照組的表達水平(Plt;0.01)。離體再灌注期間供肝組織中p38MAPK活性與供肝組織內TNF-α mRNA的表達水平呈顯著性正相關(r=0.996,Plt;0.01)。結論:p38MAPK對TNF-α生成的調節作用層次可能在轉錄水平,提示p38MAPK信號轉導途徑對TNF-α mRNA的調節可能是導致離體肝臟缺血再灌注損傷的重要機制之一。
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.
目的考察皮下通道型膽囊肝膽管成形術（STHG）治療肝膽管結石及膽管狹窄的中、遠期療效。方法對該院1994年12月至2000年6月期間行STHG手術的59例患者的術后中、遠期并發癥進行統計分析。結果STHG的術后并發癥發生率較低，而且并發癥的種類也較少； 本組病例術后無返流性膽管炎的表現，也無胃腸道功能紊亂和吻合口潰瘍發生。結論STHG既保存了膽囊及Oddi括約肌功能，又保證了膽汁的生理流向，還能防止腸液的返流，從而避免了術后消化功能紊亂和返流性膽管炎的發生，是一種較為理想的治療肝膽管結石和肝門部膽管狹窄的術式。