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.
Objective To discuss the way of animal model building of hepaticocholedochostomy(HC) and hepaticojejunostomy(HJ) and to compare the short-term effect. Metheds Twenty-nine dogs were divided randomly into control group(n=5) and the experimental group (stenosis of left hepatic duct, n=24). After 7 weeksof stenosis of left hepatic duct,24 dogs in the experimental group were divided randomly into HC subgroup (n=12) and HJ subgroup (n=12) .The operation time and the blood loss during operation were recorded and the hepatic function was detected.Results The diameter of left hepatic duct was significantly expended after 7 week’s stenosis. Hepaticocholedochostomy took shorter time and lost less blood than hepaticojejunostomy. The dogs in HC subgroup lost less weight than thosein HJ subgroup. In HC and HJ subgroups, the mortality rates were 1/12 and 3/12;the infectious rates of incision were 3/12and 5/12 respectively. Serum levels of total bilirubin and transaminase increased significantly in the 7th week after stenosis of left hepatic duct compared with before stenosis of left hepatic duct. However, Serum levels of total bilirubin and transaminase restored to normallevels after 1 month of HC or HJ.Conclusion It is feasible to establish animal model of bile duct reconstruction on the basis of stricture of bile duct. The dogs undergoing hepaticocholedochostomy have less trauma, better results than the dogs undergoing hepaticojejunostomy. Both hepaticocholedochostomy and hepaticojejunostomy are able to relieve the obstruction of bile duct.
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 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.
In recent years, the importance of guanylate cyclase-C (GC-C) in digestive system diseases has received more and more attention. It plays a key role in regulating water and electrolyte balance, maintaining gastrointestinal function, relieving abdominal pain, controlling inflammation, regulating intestinal microecology, inhibiting tumor growth and regulating cell proliferation, and is considered as a potential therapeutic target for digestive tract diseases. This article reviews the role of GC-C in digestive system diseases and related intervention studies of agonists linaclotide and plecanatide in recent years, in order to better understand its intrinsic function and further guide the diagnosis and treatment of related clinical diseases.
Objective To study the protective effects of pre-storing glycogen on warm ischemia reperfusion injury during partial hepatectomy. Methods Thirty-eight patients were randomly divided into a trial group (n=19) and a control group (n=19). In the trial group, patients were given high concentration glucose intravenously during the 24 hours before the operation. The hepatic lesion was resected after portal triad clamping in the two groups. Liver function of all patients was measured before the operation and the first and fifth days after the operation. Normal hepatic tissue was biopsied to measured hepatic tissue glycogen contents before the operation and the change of superoxide dismutase (SOD) at the point of pre-ischemia, post-ischemia, and reperfusion 2 hour. Bcl-2 mRNA, a well known anti-apoptotic factor, was also detected using quantitative polymerase chain reaction. Results The hepatic tissue glycogen content of the trial group was significantly higher than that of the control group before the operation (Plt;0.01). Liver function of the trial group was significantly better than that of the control group on the first and fifth day after operation (Plt;0.05). There was significant difference in SOD activity between the two groups at the end of hepatic vascular occlusion and at the point of 2-hour reperfusion (Plt;0.05). Furthermore Bcl-2 mRNA expression of the trial group was notably up-regulated at the point of 2-hour reperfusion compared to the control group. Conclusion Pre-store storing glycogen might protect liver ischemia reperfusion injury caused by hepatic vascular occlusion during partial hepatectomy. The potential mechanism might be that pre-storing glycogen enhances Bcl-2 expression.
ObjectiveTo evaluate the efficiency of the spot-welding electrocoagulation with needle-knife to prevent bleeding after endoscopic sphincterotomy (EST). MethodsThe clinical data of 187 patients underwent EST from August 2009 to October 2009 were retrospectively analyzed, study group （n=102) were treated with spotwelding electrocoagulation with needleknife and 110 000 noradrenaline washing, control group (n=85) were treated with 110 000 noradrenaline washing alone. The bleeding and complications after EST were observed. ResultsThe differences of gender, age, primary diseases, cormorbidities, nutritional status, and immune function were not significant between two groups (Pgt;0.05). The bleeding after EST happened 4 cases (4.70%) in the control group and none in the study group. The bleeding rate of the study group was significantly lower than that of the control group (Plt;0.05). The bleeding cases in the control group were controlled successfully by spotwelding electrocoagulation with needleknife under endoscopy. Cholangitis occurred in 5 cases altogether, 1 case in each group deteriorated promptly and died of multiple organ failure syndrome, another 3 cases, 2 in the study group, 1 in the control group, were cured by PTCD and antibiotics. Biliary tract hemorrhage occurred one case in each group, which one died in the study group. Pancreatitis occurred 1 case in the study group and 2 cases in the control group, all of which were salvaged by conservative therapy. The incidences of complications were not significantly different between two groups (Pgt;0.05). ConclusionsThe spotwelding electrocoagulation with needleknife can significantly reduce the bleeding rate after EST. It is an effective, safe, and easy technique, especially to rural areas.
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.