ObjectiveTo summarize the research progress of severed limb preservation by perfusion and to analyze difference in effect of severed limb preservation by different perfusate.
MethodsThe domestic and foreign related literature about severed limb preservation by perfusion was extensively reviewed and analyzed.
ResultsCurrently the main perfusate includes organ perfusate,free radical scavengers,energy mixture,blood substitutes,and whole blood.They can reduce the skeletal muscle's ischemia-reperfusion injury in different degrees.
ConclusionDifferent perfusate can reduce the skeletal muscle's ischemia-reperfusion injury in different degrees,but the best effect of perfusate and personalized preservation method need further study.
Objective To review the treatment methods and techniques of ischemia-reperfusion injury of flap. Methods Recent basic research l iterature concerning ischemia-reperfusion injury of flap was reviewed and analyzed in terms of treatment techniques. Results Ischemia-reperfusion injury is one of the leading causes of flap necrosis postoperatively. Interventions against any l ink of the ischemia-reperfusion injury progress could effectively reduce the damageand improve the survival rate of flaps. Conclusion Including production of reactive oxygen species, neutrophil infiltrationetc are thought to be the main mechanisms of ischemia-reperfusion injury. Treatment including medicine administration and physical intervention against any specific l ink of ischemia-reperfusion injury can interfere or block the whole progress, which reduce the damage of ischemia-reperfusion injury and improve the survival rate of animal flap models eventually.
Objective To decrease the operative difficulty, with the purpose of looking for an orthotopic liver autotransplantation model which not only materializes the liver transplantation but also possesses higher survival rate. Methods This model was established via portal vein perfusion in thirty rats, and from which the result of the liver after perfusion, the operative time and the survival rate were observed. Liver tissues were researched at 24 h after operation under the light microscope. Results This model was easy to be perfused, the operative time was (48±3.0) min and the survival rate was 96.7% (29/30). The structure of hepatic tissue was basically normal with a little hydropic degeneration under the light microscope. Few erythrocytes residual occurred in the interlobular arteries under the light microscope. Conclusion The orthotopic liver autotransplantation model via portal vein perfusion has an exclusively blockage pattern which possesses a higher survival rate. It prevents the injury of immunological rejection and purely reflects the hepatic ischemia-reperfusion. But it is better to be applied in the non-hepatic artery anastomosis or the research nothing to do with the hepatic artery because the hepatic artery does not have sufficient perfusion.
Objective To study the efect of IH764-3 on ischemia-reperfusion (I/R) injury in rat liver. Methods Rats were divided into 3 groups, the control group was not subjected to ischemia and no treatment was given. I/R injury group was subjected to 40 minutes ischemia followed by reperfusion for 120 minutes. The IH7643 group (40mg/kg) was administred at ischemia and reperfusion. Results In the IH764-3 group, sereum levels of ALT, AST, AKP and γ-GT were significantly lower than those in the I/R group. Energy charge level recovery was significantly higher with IH7643 (P＜0.05), hepatic ultrastructure was better preserved with IH764-3. Conclusion IH764-3 may be useful in the treatment of hepatic ischemia reperfusion injury
【Abstract】 Objective To study the effects of ischemic preconditioning (IP) on the activity of nuclear factor-κB （NF-κB） and the expressions of TNF-α and intercellular adhesion molecule-1 (ICAM-1) during early reperfusion following liver transplantation in rats. Methods The models of rat orthotopic liver transplantation were established. The donor livers were stored for 2 hours in Ringers solution at 4 ℃ before transplantation. All rats were randomly divided into sham operation group （SO group）， control group and IP group. IP group was achieved by clamping the portal vein and hepatic artery of donor liver for 10 minutes followed by reperfusion for 10 minutes before harvesting. The activity of NF-κB and expressions of TNF-α and ICAM-1 at 1 h, 2 h, 4 h and 6 h after reperfusion were measured. Serum ALT, LDH were also determined. Results The liver function of recipients with IP were significantly improved. Compared with SO group， the graft NF-κB activity increased after transplantation in control group and IP group (P＜0.05), while compared with control group that was significantly attenuated at 1 h and 2 h in IP group. Similarly, hepatic levels of TNF-α and ICAM-1 were significantly elevated in control group and were reduced in IP group. Conclusion IP might down-regulated TNF-α and ICAM-1 expression in the grafts after orthotopic liver transplantation through depressed NF-κB activation， and attenuate neutrophil infiltration in the grafts after reperfusion.
【Abstract】 Objective To investigate the effect of verapamil on apoptosis, calcium and expressions of bcl-2 and c-myc of pancreatic cells in ischemia-reperfusion rat model. Methods Wistar rats were randomly divided into three groups: control group （n=10）; ischemia-reperfusion group （n=10）; verapamil treatment group （n=10）. The anterior mesenteric artery and the celiac artery of rats in both ischemia-reperfusion group and verapamil treatment group were occluded for 15 min followed by 12-hour reperfusion. Verapamil （1 mg／kg） was injected via caudal vein to the rats in verapamil treatment group 15 min before occlusion and 1 hour after the initiation of reperfusion, respectively; and ischemia-reperfusion group was given the same volume of salient twice intravenously. Pancreatic tissues were collected from the dead rats after twelve hours since the reperfusion. The pathologic characters of pancreatic tissue were observed under light microscope; The level of calcium in the tissue was measured by atomic absorption spectrometer; TUNEL was used to detect apoptosis of pancreatic cells; and the expressions of c-myc and bcl-2 in the cells were also analyzed by immunohistochemistry technique and flow cytometry. Results The pathologic change in verapamil treatment group was less conspicuous than that of ischemia-reperfusion group. Both the calcium level and the number of apoptotic cells in verapamil treatment group were less than those of ischemia-reperfusion group 〔（411.1±55.8） μg/g dry weight vs （470.9±31.9） μg/g dry weight, P＜0.05 and （9.5±2.9）% vs （18.4±3.1）% 〕, P＜0.05. After taking verapamil, the number of apoptotic cells decreased, whereas the expressions of bcl-2 and c-myc increased. The fluorescent indexes of bcl-2 and c-myc in verapamil treatment group were significantly higher than those of ischemia-reperfusion group (1.72±0.11 vs 1.41±0.07, P＜0.05; 1.76±0.19 vs 1.55±0.13, P＜0.05. Conclusion Ischemia-reperfusion injury can induce apoptosis of pancreatic cells. Verapamil could protect the injured pancreatic tissue by reducing the level of calcium, stimulating the expressions of bcl-2 and c-myc and inhibiting apoptosis of pancreatic cells.
Objective To observe the protective effects of diazoxide-preconditioning on myocardial ischemiareperfusion injury of rats and discuss its possible mechanisms. Methods Fourteen healthy SD rats were randomly divided into two groups（7 each group）,In diazoxide-preconditioning group diazoxide was injected with the dosage of 12.5mg/kg through the vein,and in control group the media with the same amount was only given before ischemia. The left anterior descending branch was ligated for 2 hours. The heart was quickly excised after 2 hours reperfusion to be used for measurement of the quantity of malondialdehyde（MDA）, the activity of superoxide dismutase （SOD）, the size of myocardial infarct area, and the cell apoptosis and ultrastructure in ischemic area. Results Compared with the control group, the quantity of MDA,the percentage of the weight of myocardial infarct area/ischemic area, and the rate of cell apoptosis in the diazoxide-preconditioning group were greatly reduced （P〈0.05, 0. 01）. The damage of cell uhrastructure was obviously alleviated,Conclusion Diazoxide-preconditioning provides evident cardioprotective effect on the myocardial ischemia-reperfusion injury of rats.
To investigate the effect of propofol intra-aortic and intravenous infusion on the concentration of propofol for an ischemia-reperfusion spinal cord injury in rabbits. Methods Forty-six healthy adult New Zealand white rabbits were randomly divided into 3 groups: sal ine infusion group (group N, n=10), propofol intra-aortic infusion group (group A, n=16) and propofol intravenous infusion group (group V, n=16). The infrarenal abdominal aorta was occluded for 30 min during which propofol 50 mg/kg was infused continuously intra-aortic or intravenous with a pump in group A and V. In group N, the same volume of normal sal ine was infused in the same way and at the same rate as in group A. Upon reperfusion, propofol concentration of the spinal segments of L4-6 and T6-8 was examined in group A and V. At 48 hoursafter reperfusion, the neurological outcomes were recorded in each group. Results Mean blood pressure in group V from the time of 5 minutes after occlusion decreased more than in group N (P lt; 0.05) and than in group A from the time of 10 minutes after occlusion(P lt; 0.05). The mean blood pressure in group N increased more than in group A from 15 minutes after occlusion (P lt; 0.05). The heart rate increased more in group V from 10 minutes after occlusion than in group N and A (P lt; 0.05) in which no difference was observed. The propofol concentration in L4-6 of group A (26 950.5 ± 30 242.3) ng/g was higher than that in T6-8 of group A (3 587.4 ± 2 479.3) ng/g and both L4-6 (3 045.9 ± 2 252.9) ng/g and T6-8 (3 181.1 ± 1 720.9) ng/g of group V(P lt; 0.05). The paraplegia incidence was lower (30%) and the median of normal neurons was higher (8.4) in group A than in group N (80%, 2.2) and group V(100%, 1.9), (P lt; 0.05). There was no significant difference in group N and V in paraplegia incidenceand the median of normal neurons (P gt; 0.05). Conclusion Intra-aortic infusion shows a better neurological outcome than intravenous infusion and could contribute to higher concentration of propofol in the ischemia spinal cord.
Objective Isoflurane has an acute preconditioning effectiveness against ischemia in kidney, but this beneficial effectiveness can only last for 2-3 hours. To investigate whether isoflurane produces delayed preconditioningagainst renal ischemia/reperfusion (I/R) injury, and whether this process is mediated by hypoxia inducible factor 1α(HIF- 1α). Methods A total of 52 male C57BL/6 mice were randomly assigned to 4 groups (n=13 in each group): the controlgroup (group A), PBS/isoflurane treated group (group B), scrambled small interference RNA (siRNA)/isoflurane treated group (group C), and HIF-1α siRNA/isoflurane treated group (group D). In groups C and D, 1 mL RNase-free PBS containing 50 μg scrambled siRNA or HIF-1α siRNA was administered via tail vein 24 hours before gas exposure, respectively. Equivalent RNasefree PBS was given in groups A and B. Then the mice in groups B, C, and D were exposed to 1.5% isoflurne and 25%O2 for 2 hours; while the mice in group A received 25%O2 for 2 hours. After 24 hours, 5 mice in each group were sacrificed to assesse the expressions of HIF-1α and erythropoietin (EPO) in renal cortex by Western blot. Renal I/R injury was induced with bilateral renal pedicle occlusion for 25 minutes followed by 24 hours reperfusion on the other 8 mice. At the end of reperfusion, the serum creatinine (SCr), the blood urea nitrogen (BUN), and the histological grading were measured. Results The expressions of HIF-1α and EPO in groups B and C were significantly higher than those in group A (P lt; 0.01). The concentrations of SCr and BUN in groups B and C were significantly lower than those in group A, as well as the scores of tubules (P lt; 0.01), and the injury of kidney was amel iorated noticeably in groups B and C. The expressions of HIF-1α and the concentrations of SCr and BUN in group D were significantly lower than those in group A (P lt; 0.01). Compared with groups B and C, the expression of HIF- 1α and EPO in group D decreased markedly (P lt; 0.01), the concentrations of SCr and BUN were increased obviously, as well asthe scores of tubules (P lt; 0.01), and the renal injury was aggratived significantly. Conclusion Isoflurane produces delayed preconditioning against renal I/R injury, and this beneficial effectiveness may be mediated by HIF-1α.
Objective To investigate the protective effects of liposome prostaglandin E1（Lipo-PGE1） on myocardial ischemia-reperfusion injury （MIRI） during cardiopulmonary bypass （CPB）. Methods Thirty-two patients with clearly diagnosed heart valve disease and congenital heart disease such as atria septal defect （ASD） and ventricular septal defect （VSD） were selected in our hospital. The patients were randomly divided into two groups （16 patients in each group）, Lipo-PGE1 group： Lipo-PGE1（2ng/kg·min） was continuously pumped before starting of CPB until 2 h after ascending aortic off-clamping; control group： using the same volume of normal saline, arterial blood samples were taken before CPB, at 1, 2, 6 and 24 h after the ascending aortic off-clamping. The value of cardiac troponin I （cTnI）, creatine kinase MBmass （CK-MB）, interleukin-6 （IL-6）, tumour necrosis factor-α （TNF-α）, intercellular adhesion molecule-l（sICAM-1） were measured. Results cTnI, CK-MB, IL-6, TNF-α and sICAM-1 showed no significant difference in the two groups before CPB （P〉0. 05）. At 1,2, 6 and 24h after ascending aortic off-clamping, those values rose significantly than before CPB（P〈0. 01）, but Lipo-PGE1 group＇s values were lower than those in the control group （P〈0. 05）. Conclusions Lipo-PGE1 （2ng/kg·min） continuously pumped from before CPB to 2h after ascending aortic off-clamping can inhibit effectively the production of IL-6, TNF-α, and reduce the expression of sICAM-1, attenuate the process of inflammation, lighten the injuries of myocardial cells, and effectively protect the MIRI during CPB open heart surgeries.
