Objective To investigate the maximum tolerance limit of rats to hepatic inflow occlusion with portal vein blood bypss (PBB) in normothermia. Methods First. A new animal model was established, the animal survival rate were calculated following 7 days of reperfusion after hepatic inflow occlusion of 30, 60, 90, 100, 110, 120 min or portal triad clamping (PTC) of 30 min. And then, the hepatic energy metabolism (RCR, P/O, ATP, AKBR) was studied following 30, 90, 120 min of ischemia or 1, 6, and 24 hours of reperfusion after the ischemia. According to the reversibility of the hepatic motochondrial function injury and maximum as long as a period of liver warm ischemia of all animal postoperative 7 days survial, the safe limit of rat to hepatic inflow occlusion was evaluated. Results The survival rate on postoperative 7 days was one hundred percent subjected to 30, 60 and 90 min of hepatic inflow occlusion, and 50, 30, 20 percent in 100, 110, 120 min, respectively, the survival rate in rats with 30 min of portal triad champing was about 40 percent. The parameters of hepatic motochondrial function reflecting the degree of liver damage to ischemia showed significantly different as compared to sham group. The functional lesion was exacerbated during inital reperfusion, then was restored progressively in PBB-30 min and PBB-90 min groups, but was maintained low level in PBB-120 min and PTC-30 min groups.Conclusion The 90 minutes is the maximum limit of rats to hepatic inflow occlusion in normothermia.
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
Objective To observe the protective role of the ectogenesis zinc on the cells in rat flap with ischemia reperfusion injury and study the mechanisms. Methods A right low abdominal island flap was created in Wistar rats. Fortyeight rats were randomly divded into 3 groups (n=16):the control group, the ischemia reperfusion group and adding zinc ischemia reperfusion group.The content of malondialdehyde(MDA) and the activity of myeloperoxidase(MPO) were measured by thiobarbituric acid methods and colorimetry. The location of expression of MT was observed,and the image analysis was performed. The quantity of MT was represented by the integratial optical density. The ultrastructure changes of skin flap with ischemia reperfusion injury and the flap viability were observed. Results In the ischemia reperfusion injury flaps, the content of MDA and MPO show no statistically significant difference among the control group,IR group and the adding-zinc-IR group (P＞0.05). Compared with the control group at 1 h and 24 h of reperfusion, the level of MDA increased 62.2% and 136.4%(P＜0.01) in the IR group, which increased 11.3% and 33.2%(P＜0.01) in the adding-zinc-IR group. The activity of MPO increased 238.4% and 503.4%(P＜0.01)in the IR group when compared with the control group, and increased 17.9%and 24.1%(P＜0.05) when compared with the adding-zinc-IR group. In the ischemia reperfusion injury falps, the content of MT in the control group and the IR group is too minimal to measure. While the content ofMT in the adding-zinc-IR group is 45.30±7.60. At 1 h and 24 h of reperfusiion, the content of MT in the adding-zinc-IR group increased 41.5% and 44.9% (P＜0.01) compared with the IR group, and increased 119.9% and 234.6% (P＜0.01) compared with the control group. The flap viability is 100% in the control group, 19.65%±4.38% in the IR group, and 24.99%±5.12% in the adding-zinc-IR group, which increased 27.2% (P＜0.05) compared with IR group. Conclusion Many kinds of cells in skin flap with ischemiareperfusion injury can be protected by ectogenesis zinc and the flap viability increases significantly.
Objective To investigate the pathological changes in the neuromuscular junction during ischemiareperfusion(IR) in the skeletal muscle. Methods Forty-eight healthy adult Wistar rats (24 male, 24 female) were equally randomised into the following 6 groups: Group A (control group): no ischemiareperfusion; Group B: ischemia by clamping the blood vessels of the right hindlimb for 3 hours; Group C: ischemia by clamping for 4.5 hours;Group D: ischemia by the clamping for 4.5 hours followed by reperfusion for 1.5hours; Group E: ischemia for 4.5 hours followed by reperfusion for 24 hours; and Group F: ischemia for 4.5 hours followed by reperfusion for 2 weeks. Then, the medial head of the gastrocnemius muscle flap model was applied to the right hindlimb of each rat. The medial head of the gastrocnemius muscle was isolated completely,leaving only the major vascular pedicle, nerve and tendons intact.The proximal and distal ends (tendons) were ligated while the vessel pedicle was clamped. And then, Parameters of the muscle (performance,contraction index,colour,edema,bleeding) were observed. The muscle harvested was stained with gold chloride(AuCl3) and the enzymhistochemistry assay (succinate dehydrogenase combined with acetylcholine esterase) was performed. Morphology and configuration of the neuromuscular junction were observed during the ischemiareperfusion injury by means of the AuCl-3 staining. The result of the enzymhistochemical reactions was quantitatively analyzed with the computer imageanalysis system. And then, additional 5 rats were prepared for 3 different models identical with those in Groups A, C and E separately. The specimens were harvested from each rat and were stained with HE and AuCl-3, and they were examined under the light microscope. Results During the period of ischemia, the skeletal muscle of Group B showed the colour of purple and edema.The colour and edema became worse in Group ,while dysfunction of elasticity and contraction appeared obviously with plenty of dark red hemorrhagic effusion at the same time.After reperfusion,the color and edema of muscle in Group D became improved while the elasticity and function of contraction was not improved. Hemorrhagic effusion of Group D turned clearer and less than Group C.Group E was similar to Group D in these aspects of muscle except for much less hemorrhagic effusion. Skeletal muscle in Group F showed colour of red alternating with white, adhesion,contracture of muscle, exposure of necrotic yellow tissue and almost lost all its functions. The AuCl3 staining showed that during IR, necrosis of the myocytes was followed by degeneration of their neuromuscular junctions, and finally the nerve fibers attached to these neuromuscular junctions were disrupted like the withering of leaves. The enzymhistochemistry assay showed thatthere was no significant difference in the level of acetylcholine esterase between the ischemic group (Groups B and C) and the control group (Group A) (Pgt;0.05). However, the level of acetylcholine esterase in all the reperfused groups (Groups D, E and F) decreased significantly when compared with the control group（Group A）and the ischemic groups (Groups B and C) (Plt;0.01). Conclusion The distribution of the nerve fibers and the neuromuscular junctions in the mass of the muscles is almost like the shape of a tree. The neuromuscular junction seems to be more tolerant for ischemia than the myocyte. Survival ofthe neuromuscular junction depends on its myocytes alive. Therefore, an ischemiareperfusion injury will not be controlled unless an extensive debridement of the necrotic muscle is performed.
【Abstract】ObjectiveOn the basis of traditional transplantation model, a successful model of pancreaticoduodenal transplantation (PDT) were established in rats, which is the foundation of basic and clinical transplantation research. Methods We improved the technique of microoperation on donor and harvested high-quality graft. The dual cuff technique was applied to end-to-end anastomose proximal part of abdominal aorta and portal vein with left renal aorta and vein of recipient, and distal part of abdominal aorta was connected with Y-tube. External secretion was performed by duodenum stoma. The PDT model was finished without blocking systemic circulation and portal vein system. Random blood glucose levels and drainage were monitored postoperatively to evaluate the function of endocrine and ectocrine. Results Thirty operations were done. The total procedure of transplantation lasted 2 hours. Moreover the operation on recipient and the reconstruction of vessels took only (26±5) and (25±5) minutes, respectively. The success rate was elevated to 100%. The ectocrine function was restored within 2 hours after operation. Except for 3 cases of non-function graft because of thrombosis in cannula, the glucose level of the remaining recipients was reduced to normal level 6 h or 24 h after transplantation. The survival rate of graft function was 90% (27/30). Conclusion This model is finished without special equipment and can recover the endocrine function in advance. It is a simple and stable model, which might be used in research of the theoretical problems involved in clinical pancreas transplantation.
To investigate the protective effect of propofol on ischemia/reperfusion induced spinal cord injury in rabbits and its influence on excitatory amino acid (EAA). Methods Sixty New Zealand white rabbits weighing 2.0-2.5 kg, half males and half females, were selected. The infrarenal circumaortic clamping model was used. And 6 mL/kg different fluids were continuously infused through a catheter into the aorta distal to the clamping site at a speed of 12 mL/(kg•h) during the 30 minutes ischemia period. According to the different infusing l iquids, the rabbits were randomized into 6 groups(n=10 per group): group A, normal sal ine; group B, 10% intral ipid; group C, propofol 30 mg/kg; group D, propofol 40 mg/kg; group E, propofol 50 mg/kg; group F, propofol 60 mg/kg. At 0, 6, 24, and 48 hours after reperfusion, neurologic outcomes were scored on a Tarlov scale system. At 48 hours after reperfusion, the number of normal neurons in the anterior spinal cord was counted, and concentration of EAA in the lumbar spinal cord was measured by high performance l iquid chromatography. Results The neuroethological score was better in groups C, D, E and F than that of groups A and B (P lt; 0.05), the score of group E was the highest (P lt; 0.05), and there was no significant difference between group A and group B (P gt; 0.05). The number of normal neurons in the anterior spinal cord of groups C, D, E and F was greater than that of groups A and B (P lt; 0.05), and group E was greater than groups C, D and F (P lt; 0.05). The concentration of EAA in groups A, B, C, D, E and F was greater than that of normal tissue, the group E was the lowest (P lt; 0.05), the groups A and B were the highest (P lt; 0.05), and there was no significant difference between group A and group B (P gt; 0.05). Concentrations of glutamate and aspartic acid were negatively correlated to normal neuron numbers in the anterior spinal cord and neuroethological scores 48 hours after reperfusion, and the corresponding correlation coefficient was — 0.613, — 0.536, — 0.874 and — 0.813, respectively (P lt; 0.01). Conclusion Propofol can significantly inhibit the accumulation of EAA in spinal cord and provide a protective effect against the ischemia/reperfusion injury induced spinal cord in rabbits.
Objective To evaluate the effect of regiono-perfusional chemotherapy of pancreatic adenocarcinoma, and to seek the management of its complications. MethodsThirty-six patients with unresectable pancreatic adenocarcinoma received selectively intra-arterial catheterization and perfused with 5-Fu, ADM, DDP. Results Six patients had complete response, 15 partial response, and one underwent radical resection subsequently. Cmplications occurred in 14 patients with 2 patients died of complications.Conclusion Regiono-perfusional chemotherapy of pancreatic adenocarcinoma is effective, but the complications can not be neglected.
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.
Objective To analyze the effects of different perfusion and different superior hepatic vena cava anastomosis methods on the establishment of rat orthotopic liver transplantation model. Methods Eighty SD rats were randomly grouped with donor group and recipient group with 40 rats in each group. Ten rats in the donor liver acquisition group received abdominal aorta infusion set drip irrigation (1 drop/s); 10 rats received abdominal aorta micro-pump perfusion method (6 mL/min); and then received recipient liver transplantation (corresponding to 10 rats in each group). The donor liver perfusion time and donor liver acquisition time of rats in the 2 groups were collected. HE staining was performed on liver tissues after perfusion and 24 h after liver transplantation. Ten rats in the recipient liver transplantation group received continuous anastomosis, and 10 rats received tension-reducing half-needle anastomosis. We collected the anastomosis time of superior and inferior hepatic vena cava, the time of anhepatic stage and the incidence of postoperative complications of the recipient. Results Contrasted with abdominal aorta infusion group, the perfusion time and acquisition time of donor liver were shorter in the abdominal aorta micro-pump perfusion group (P＜0.05). HE staining showed that the morphology of hepatocytes, portal vein and bile duct in the abdominal aorta micro-pump perfusion group did not change obviously, only a few lymphocytes infiltrated. Contrasted with continuous anastomosis group, the anastomosis time, anhepatic stage of the superior hepatic vena cava, incidences of postoperative anastomotic bleeding and incomplete perfusion of donor liver in the reduced tension half-needle anastomosis group were shorter or lower (P＜0.05). Conclusions Compared with abdominal aorta infusion set drip method, the quality of donor liver was improved by abdominal aorta micropump perfusion. Compared with continuous anastomosis method, the tension-reducing half-needle anastomosis can shorten the suture time of superior hepatic vena cava and anhepatic stage, and the incidence of anastomotic bleeding was reduced.
【Abstract】Objective To investigate the anatomic structure and experimental methods of peribiliary vascular plexus(PVP) in rat’s hepatic portal bile duct.Methods To observe the morphological structure of PVP in rat’s hepatic portal bile duct after the hepatic artery was perfused with Chinese ink and transparency management, performed and to make three dimensional reconstruction of PVP’s spacial structure in MoticBuaa3Dvol software.Results The microvascular distribution and plane structure of PVP in rat’s hepatic portal bile duct could be shown clearly through Chinese ink perfusion and transparency management. The three dimensional structure of PVP could be reconstructed effectively by MoticBuaa3Dvol software, its effect was verisimilitude. Conclusion Chinese ink perfusion and vitrification is a simple and easy method in PVP’s morphologic study of rat’s hepatic portal bile duct. MoticBuaa3Dvol software is useful in microvascular study of hepatic portal bile duct.