Objective To investigate the protective effects of antitumor necrosis factor-α antibody （TNF-αAb） on lung injury after cardiopulmonary bypass （CPB） and their mechanisms. Methods Forty healthy New Zealand white rabbits，weighting 2.0-2.5 kg，male or female，were randomly divided into 4 groups with 10 rabbits in each group. In groupⅠ，the rabbits received CPB and pulmonary arterial perfusion. In group Ⅱ，the rabbits received CPB and pulmonary arterial perfusion with TNF-αAb. In group Ⅲ，the rabbits received CPB only. In group Ⅳ，the rabbits only received sham surgery. Neutrophils count，TNF-α and malondialdehyde （MDA） concentrations of the blood samples from the left and right atrium as well as oxygenation index were examined before and after CPB in the 4 groups. Pathological and ultrastructural changes of the lung tissues were observed under light and electron microscopes. Lung water content，TNF-α mRNA and apoptoticindex of the lung tissues were measured at different time points. Results Compared with group Ⅳ，after CPB，the rabbitsin group Ⅰ to group Ⅲ showed significantly higher blood levels of neutrophils count，TNF-α and MDA（P＜0.05），higherTNF-α mRNA expression，apoptosis index and water content of the lung tissues （P＜0.05），and significantly lower oxyg-enation index （P＜0.05） as well as considerable pathomorphological changes in the lung tissues. Compared with group Ⅱ，after CPB，the rabbits in groups Ⅰ and Ⅲ had significantly higher blood concentrations of TNF-α （5 minutes after aortic declamping，220.43±16.44 pg/ml vs.185.27±11.78 pg/ml，P＜0.05；249.99±14.09 pg/ml vs.185.27±11.78 pg/ml，P＜0.05），significantly higher apoptosis index （at the time of CPB termination，60.7‰±13.09‰ vs. 37.9‰±7.78‰，P＜0.05；59.6‰±7.74‰ vs. 37.9‰±7.78‰，P＜0.05），significantly higher blood levels of neutrophils count and MDA （P＜0.05），significantly higher TNF-α mRNA expression and water content of the lung tissues （P＜0.05），and significantly loweroxygenation index （P＜0.05） as well as considerable pathomorphological changes in the lung tissues. Compared with groupⅠ，rabbits in group Ⅲ had significantly higher above parameters （P＜0.05） but lower oxygenation index （P＜0.05） only at 30 minutes after the start of CPB. Conclusion Pulmonary artery perfusion with TNF-αAb can significantly attenuate inflammatory lung injury and apoptosis of the lung tissues during CPB.
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 investigate the changes of expression of zonula occludens-1(ZO-1) in rats with severe acute pancreatitis (SAP), and to study the relationship between the ZO-1 protein and microvascular injury in rats with SAP. Methods Forty-eight Wistar rats were randomly divided into sham-operation (SO) group and SAP group, each group enrolled 24 rats. Pancreas of rats in SO group were flipped only after laparotomies, but rats of SAP group were injected with 5% sodium taurocholate by retrograding cholangiopancreatography micro pump to produce the SAP model. At 6, 12, and 24 hours after operation, 8 rats were sacrificed to get abdominal aortic blood for testing the levels of peripheral blood amylase, trypsin, interleukin-8(IL-8), tumor necrosis factor-α(TNF-α), and ZO-1 protein. At the same time, pancreatic tissues were got to perform HE staining and immunohistochemical staining for observation of the pathological changes and the expression of ZO-1 protein respectively. Results Compared with SO group at the same time, the levels of peripheral blood amylase, trypsin, IL-8, TNF-α, and ZO-1 protein were all higher in SAP group (P < 0.05). The level of amylase in SAP-24 hours group was higher than those of 6 hours group and 12 hours group(P < 0.05), the levels of trypsin, IL-8, and ZO-1 protein in SAP group increased over time (P < 0.05), but levels of TNF-αin 3 time points of SAP group did not differ with each other significantly(P > 0.05). Results of regression showed that in the SAP group, the level of ZO-1 protein in serum was significantly positive correlated with pathological score of pancreatic tissue(b=0.96, P < 0.05), levels of serum amylase(b=0.87, P < 0.05), trypsin(b=0.72, P < 0.05), and serum IL-8 (b=0.69, P < 0.05), but was not significantly correlated with level of TNF-α(P > 0.05). HE staining results showed that damage of pancreatic tissues became worse over time in SAP group, and the pathological score of SAP-6 hours group was lower than those of 12 hours group and 24 hours group (P < 0.05). Immunohistochemical staining results showed that, in SAP group, with the extension of time, the number of ZO-1 protein granules in pancreatic acinar cells and capillary wall reduced, and expressed in capillaries discontinuously. Conclusion During the course of SAP, the concentration of serum ZO-1 protein increase, but its expression in the pancreatic tissue degrade, which is closely associated with microvascular injury and progression of pancreatic tissues.
The damage effects of the pure tumor necrosis factor (TNF) on the normal animals were observed. Eighteeen rabbits were divided into two groups, eight in tested group and ten in control group. 0.5mg per kg of the pure rabbit TNF was given to each animal of the tested group. Results:The symptoms similar to that induced by endotoxin appeared after the TNF injection. The functions of the main organs were markedly damaged. The arterial blood pressure of most animal was low. The weight ratio of the orgen to the body was raised. The pathologic changes were similar to those of the multiple organ failure (MOF) model. Most of the animal died before the end of the experiment. The results suggest that pure TNF could indece multiple organ damages similar to those of MOF.
目的:分析脑出血死亡时间及死因,找出脑出血不同死亡时间的主要死因,为制定脑出血不同时间的相应救治措施提供依据。方法:回顾性总结分析２１１ 例脑出血死亡病例的死亡时间、主要死因,找出不同死亡时间对应的主要死因。 结果: (１)死亡时间:≤３ 天 ９１ 例、４~７天 ５２ 例、８~１４ 天 ４２ 例、１５~２１ 天１８ 例、２２~２８ 天 ７ 例、≥ ２９ 天 １ 例。(２)死亡原因:１６９ 例死于脑疝及中枢性呼吸、循环衰竭;20例死于肺部感染;１０ 例死于消化道出血;4 例死于多器官功能衰竭;４ 例死于心脏病变(如心脏猝死、心肌梗塞、心功能衰竭);其它４例(痰窒息、肾功能衰竭、肝功能衰竭)。(３)不同死亡时间主要死亡原因:≤３ 天 ９０ 例死于脑疝及中枢性呼吸、循环衰竭,仅 １ 例死于消化道出血;４～７ 天４７ 例死于脑疝及中枢性呼吸、循环衰竭,３ 例死于肺部感染,死于心脏病变及其它各１例;８~１４ 天 ２９ 例死于脑疝及中枢性呼吸、循环衰竭,８ 例死于肺部感染,２ 例死于消化道出血,死于心脏病变、多器官功能衰竭、其它各 １ 例;１５~２１ 天 ７ 例死于肺部感染,５ 例死于消化道出血,３ 例死于脑疝及中枢性呼吸、循环衰竭,死于心脏病变、多器官功能衰竭、其它各１例;２２～２８天死于消化道出血、肺部感染各２ 例,死于心脏病变、多器官功能衰竭、其它各１ 例,已无死于脑疝及中枢性呼吸、循环衰竭者;２９ 天后死于多器官功能衰竭１ 例。结论: 脑出血死亡时间不同,其主要死亡原因不同。临床应该针对不同死亡时间的主要死因制定相应救治方案,以降低死亡率。
OBJECTIVE: To investigate the protective effect of tumor necrosis factor-alpha(TNF-alpha) on spinal motor neurons after peripheral nerve injury. METHODS: Twenty Wistar rats were divided into two groups, the right sciatic nerves of 20 Wistar rats were transected, the proximal stumps were inserted into a single blind silicone tube. 16 microliters of normal saline(NS) and TNF-alpha(30 U/ml) were injected into the silicone tubes. After 2 weeks, the 4th, 5th lumbar spinal cord were taken for examination. Enzyme histochemical technique and image analysis were used to show acetylcholinesterase(AChE) and nitric oxide synthase(NOS) activity of spinal motor neurons. RESULTS: The number of AChE and NOS staining neurons were 8.65 +/- 1.98 and 5.92 +/- 1.36 in the experimental group and 6.37 +/- 1.42 and 8.67 +/- 1.45 in the control group respectively, there were significant difference between the two groups(P lt; 0.01). CONCLUSION: It suggests that TNF-alpha has protective effect on motor neurons after peripheral nerve injury.
Objective To investigate the protective effects of endotoxin pretreatment on lung injury of rats with endotoxemia. Methods The rat model of acute endotoxemia was established by injecting lipopolysaccharide (LPS) intraperitoneally. Seventy-two male Wistar rats were randomly divided into three groups, ie. a saline control group (N, n=24) , a LPS-treated group (L, n=24) , and a LPS pretreated group ( P, n=24) . Each group was divided into 2 h, 4 h, 6 h, and 12 h subgroups. The rats in group P were firstly administered with introperitoneal injection of 0.25 mg/kg LPS. After 24 hours, they were subjected to the injection of 0.5 mg/kg LPS. The rats in group N and L received injection of equivalent amount of saline. After 72 hours, the rats in group L and P were challenged with intravenous injection of 10 mg/kg LPS, otherwise saline in group N. Six rats were killed at 2, 4, 6 and 12 hours respectively after injection of LPS in group L and P. The lungs were removed for detecting intercellular adhesion molecule-1 ( ICAM-1) , superoxide dismutase ( SOD) , and malondialdehyde (MDA) . Meanwhile the level of tumor necrosis factoralpha ( TNF-α) in serum was measured, and the pathological changes of lung were also examined. Results The contents of ICAM-1, MDA and TNF-α in the LPS-treated 4 h group were 75.07 ±0. 53, ( 3.93 ± 0.42) μmol/g, and (478.62 ±45.58) pg/mL respectively, significantly higher than those in the saline control group. The endotoxin pretreatment reduced the above indexes to 42.40 ±0.44, ( 2.89 ±0.49) μmol / g and ( 376.76 ±43.67) pg/mL respectively (Plt;0.05) . The content of SOD in the LPS-treated 4 h group was ( 6.26 ±0.31) U/mg, significantly lower than that in the saline control group. The endotoxin pretreatment increased SOD to ( 8.79 ±0.35) U/mg. Conclusion Endotoxin pretreatment can suppress the progress of lung injury in rats with endotoxemia and protect the lung tissue by down-regulating the inflammatory response and oxygen free radical production.
In order to observe activity of tumor necrosis factor (TNF) in the serum, pancreatic histopathological damage, as well as their relationships in acute necrotizing pancreatitis (ANP), thirty five SD rats were randomly divided into 7 groups according to their sampling time with 5 in each group. ANP was induced by retrograde infusion of 5% sodium taurocholate through biliopancreatic duct in 6 experimental groups (Group B1～B6).Blood and pancreatic tissue samples were obtained at hour 0,0.5,2,4,6 or 8 respectively when the animals were sacrificed.Results showed that serum level of TNF activity rose significantly in Group B2,and reached the maximal value in Group B4.The pancreatic histopathological damage in ANP rats was getting worse along with time. Serum TNF activity had close relation to pancreatic histopathological score (r=0.63, Ｐ＜0.01),suggesting that serum TNF may play an important role in the process of deterioration of pancreatic tissue damage during ANP.