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ObjectiveTo explore the influence of Xiaoai Jiedu prescription （XJP）-containing serum on natural killer （NK） cells′ lethal effect on colon cancer cells and the molecular mechanism. MethodXJP-containing serum （0.1%， 0.5%， 1%， 5%， 10%） was used to treat HCT-116 cells and NK-92MI cells respectively for 24 h， and methyl thiazolyl tetrazolium （MTT） assay was employed to detect cell proliferation. Then， low-concentration （0.1%， 0.5%， 1%） XJP-containing serum was selected to treat co-cultured HCT-116 cells and NK-92MI cells for 24 h and calcein acetoxymethyl ester/propidium iodide （Calcein-AM/PI） was applied to detect the killing effect of NK cells on colon cancer cells. Flow cytometry was used to detect apoptosis of colon cancer cells， Western blot the expression of apoptosis-related proteins and signal transducer and activator of transcription 4 （STAT4） pathway-related proteins， and enzyme-linked immunosorbent assay （ELISA） the secretion of interferon （IFN）-γ. ResultHigh-concentration （5%， 10%） XJP-containing serum inhibited the proliferation of HCT-116 and NK-92MI cells （P<0.01）， while low-concentration （0.1%， 0.5%， 1%） XJP-containing serum had no obvious influence on cell proliferation compared with the blank group. As compared with the blank group， low-concentration XJP-containing serum enhanced the killing activity of NK cells against colon cancer cells in a concentration-dependent manner （P<0.01）， and induced apoptosis of colon cancer cells （P<0.01）. Moreover， XJP-containing serum （0.1%， 0.5%， 1%） down-regulated the expression of B-cell lymphoma 2 （Bcl-2） and B-cell lymphoma-extra large （Bcl-xl）， and up-regulated the expression of Bcl-2-associated X （Bax） compared with the blank group （P<0.05， P<0.01）. Compared with the co-culture group， XJP-containing serum （0.1%， 0.5%， 1%） increased the expression of p-STAT4 and IFN-γ （P<0.05）. ELISA result showed that XJP-containing serum （0.1%， 0.5%， 1%） raised IFN-γ secretion （P<0.01）. ConclusionXJP-containing serum can enhance the activity of NK cells to kill colon cancer cells. The mechanism is the likelihood that it activates STAT4 pathway， increases IFN-γ secretion by NK cells， down-regulates the expression of Bcl-xl and Bcl-2， and up-regulates the expression of Bax， thereby promoting the apoptosis of colon cancer cells.
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【Objective】 To generate secreted frizzled-related protein 4 (SFRP4) transgenic mice and analyze their biological properties. 【Methods】 We generated SFRP4 transgenic mice by DNA microinjection and detected the expression of SFRP4 by qRT-PCR and Western blotting. 【Results】 SFRP4 transgenic mice were successfully generated by DNA microinjection. The expression of SFRP4 was dramatically increased in transgenic mice liver compared with that of wide type. 【Conclusion】 The transgenic mice model of SFRP4 was successfully established by DNA microinjection. It provides a good model for studying the function of SFRP4 and the mechanism of participating in metabolic diseases.
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Objective To evaluate the effect of hyperoxygenated solution on myocardial injury in the rats with acute carbon monoxide (CO) poisoning.Methods Thirty pathogen-free adult male SpragueDawley rats,weighing 250-300 g,were randomly divided into 5 groups (n=6 each) using a random number table:control group (C group),acute CO poisoning group (ACP group),and different doses of hyperoxygenated solution groups (HP1-3 groups).CO 120 ml/kg was injected intraperitoneally to establish the model of acute CO poisoning.Hyperoxygenated solution 10,15 and 20 ml/kg were infused via the caudal vein at 1 h after intraperitoneal injection of CO in HP1-3 groups,respectively.At 24 h after intraperitoneal injection of CO,blood samples were collected from the caudal vein for determination of plasma creatine kinase (CK),creatine kinase-MB (CK-MB),lactic dehydrogenase (LDH) and alpha-hydroxybutyrate acid dehydrogenase (α-HBDH) activities using the automatic biochemical analyzer.The rats were then sacrificed,and myocardial specimens were obtained for examination of the pathological changes with a light microscope.Results Compared with group C,the plasma LDH,α-HBDH,CK and CK-MB activities were significantly increased in ACP and HP1-3 groups (P＜0.01).Compared with group ACP,the plasma LDH,α-HBDH,CK and CK-MB activities were significantly decreased in HP1-3 groups (P＜0.05 or 0.01).Compared with group HP1,the plasma LDH,α-HBDH,CK and CK-MB activities were significantly decreased in HP2,3 groups (P＜0.05).The pathological changes of myocardium were significantly attenuated in HP1-3 groups as compared with group ACP.Conclusion Hyperoxygenated solution can attenuate myocardial injury in the rats with acute CO poisoning.
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Objective To observe effects of propofol versus urapidil on perioperative hemodynamics in patients with gallbladder opera-tion during tracheal extubation. Methods From 2010 January to 2012 December, 128 patients who were diagnosed by color B ultra /CT and then underwent gallbladder selective operation (ASAⅡ~IV) were selected, and they were divided into the propofol group (n=64) and the urapidil group (n=64). At the end of operation, patients of the two groups were given intravenous injection of propofol 1. 5 mg/kg and ura-pidil 2. 5mg/kg which were diluted with normal saline to 8 mL respectively. Sputum suction immediately after medication, and then wiped out the endotracheal tube and gave oxygen masks for 10 min. Record the systolic/diastolic blood pressure ( SBP/DSP) , heart rate ( HR) , pH, PaO2 , PaCO2 and SaO2 under double blind trial before induction, after medication, at the time of sputum suction, at the time of extuba-tion, 5 min after extubation and 10 min after extubation. At the same time, agitation during the extubation period and patients awake time were recorded. Results After extubation, cough (4. 7% vs. 26. 6%), agitation (3. 1% vs. 17. 2%) and glossoptosis (12. 5% vs. 21. 9%) in propofol group was significantly lower than urapidil group (P0. 05). During perioperation extubation, there was no significantly difference in terms of changes of pH, PaO2, PaCO2 and SaO2 be-tween the groups (P>0. 05). Conclusion Propofol is better than urapidil in preventing adverse effect of extubation for patients with gall-bladder operation, and it will not affect the recovery time of patients.
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Objective To evaluate the effect of isoflurane preconditioning on inflammatory responses during spinal cord injury (SCI ) in rats .Methods Sixty adult male Sprague-Dawley rats ,weighing 250-300 g , were randomly divided into 3 groups ( n= 20 each ) using a random number table :sham operation group (S group) , SCI group , and isoflurane preconditioning group (I group ) . The animals were anesthetized with intraperitoneal pentobarbital sodium 40 mg/kg .SCI was produced by a weight-drop contusion at the T10 level .The rats inhaled 2% isoflurane for 2 h ,and the model was established at 24 h after the end of isoflurane inhalation in I group . Neurological function was assessed and scored by using the the Basso , Beattie , Bresnahan (BBB ) Locomotor Rating Scale on 7 days after SCI .Five rats in each group were then chosen and spinal cord specimens were obtained and cut into sections which were stained with haematoxylin and eosin for determination of the viable neuron count .Fifteen rats in each group were sacrificed and the spinal cord was removed for detection of nuclear factor kappaB (NF-κB ) and interleukin-1β (IL-1β) expression (by Western blot ) .Results Compared with S group ,BBB score and the number of viable neurons were significantly decreased ,and the expression of NF-κB and IL-1βprotein was up-regulated in SCI group ( P<0.05) .Compared with SCI group ,BBB score and the number of viable neurons were significantly increased ,and the expression of NF-κB and IL-1βprotein was down-regulated in group I ( P<0.05 ) .Conclusion The mechanism by which isoflurane preconditioning protects the spinal cord is related to inhibition of inflammatory responses in rats .
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ObjectiveTo evaluate the biological characteristics of rat bone marrow mesenchymal stem cells (BMSCs) transfected with hypoxia-inducible factor-1α(HIF-1 α) gene.MethodsThe rat BMSCs of 3rd generation and the vector expressing HIF-1α gene (pcDNA3.1-HIF-1α) were provided by department of anesthesia,Tangdu Hospital,the 4th Military Medical University.BMSCs expressing HIF-1α gene (BMSCs-HIF-1α cells) were constructed by transfection of vector pcDNA3.1-HIF-1α into BMSCs by means of electroporation.Successful transfection of HIF-1α gene was confirmed by immuno-cytochemistry.Simple BMSCs and BMSCs-pcDNA3.1 cells were used as control cells.After being cultured in hypoxic condition HIF-1α expression was detected by Western blot analysis.Flow cytometry was used to determine the proportion of cells in G1,G2 and S phase and detect apoptosis.The proliferation index (PI) was calculated.The cell growth curve was described by MTT assay and the number of the 3 types of cells was recorded.ResultsA large number of deep blue granules were observed in the nuclei of BMSCs-HIF-1α cells using immuno-cytochemistry but no such granule was found in the two types of control cells.HIF-1α expression was significantly up-regulated and apoptosis rate (the number of apoptotic cella/the total number of cells examined) decreased in BMSC-HIF-1α cells compared with the control cells.The proportion of cells in S and G2 phase was significantly higher and the proportion of cells in G1 phase was significantly lower and PI higher in BMSCs-HIF-1α cells than in the control cells.The number of BMSCs-HIF-lα cells was significantly higher than the number of the two types of control cells at day 3-8 of culture.There was no significant difference in the above variables between BMSCs and BMSCs-pcDNA3.1 cells.ConclusionBMSCs-HIF-1α is successfully constructed by transfection of vector pcDNA3.1-HIF-1α gene into BMSCs by means of electroporation.
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Objective To investigate the effects of propofol on intracellular free Ca2+ concentration and nitric oxide synthase (NOS) activity in PC12 cells exposed to bupivacaine. Methods The PC12 cells were provided by Shanghai Cell Biology Research Institute, Chinese Academy of Sciences and cultured in DMEM liquid culture medium. The cultured PC12 cells were seeded in 36 well plates and randomly assigned to one of 4 groups (n=9 wells each): group Ⅰ control (C);group Ⅱ propofol (P);group Ⅲ bupivacaine (B) and group Ⅳ propofol + bupivacaine (PB). In control group D-Hank solution was added. The cells were exposed to propofol 2 mmol/L and bupivacaine 0.09 mmol/L in group P and B respectively. In group PB the cells were incubated with propofol 2 mmol/L and bupivacaine 0.09 mmol/L simultaneously. After being incubated for 6 and 24 h the apoptosis in BC12 cells was assessed by flow cytometry. Apoptotic rate was calculated. NOS activity and intracellular free Ca2+ coneentration in PC12 cells were determined. Results Bupivacaine significantly increased the apoptotic rate of PC12 cells, the intracellular free Ca2+ concentration and NOS activity in PC12 cells in group B as compared with control group. Propofol significantly decreased the toxic effects of bupivacaine on PC12 cells in group PB compared with group B. Conclusion Bupivacaine is toxic to PC12 cells by increasing apoptosis, intracellular Ca+ concentration and NOS activity in the cells. The toxic effects can be prevented to some extent by concomitant administration of propofol.
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Objective To investigate the expression and co-existence of fos-protein and enkephalin in CNS following enflurane anesthesia. Methods Twelve adult SD rats of both sexes weighing 195-223g were divided randomly into two equal groups: control group and enflurane group: The animals in enflurane group breathed 2% enflurane for 2h. The animals in control group underwent the same experimental steps except enflurane inhalation. Before experiment the animals were kept in a quiet place for 24h and strong light was avoided. After enflurane inhalation, chest was opened and 100 ml of normal saline was infused via left ventricle to wash Out blood from whole body, then followed by infusion of 4% polymerized formaldehyde 0.1mol/L PB 500 ml for fixation of tissue. 90 mm later the whole brain and spinal cord were harvested for determination of fos-protein and enkephalin expression and their location using double-labelled immunohistochemical technique. Results The control group showed more ELI(enkephalin like immunoactivity) neurons, less FLI(fos like immuneactivity) neurons and FLI/ELI(fos and enkephalin like immunoactivity) neurons were very rare. The enflurane group showed more FLI, ELI and FLI/ELI neurons. They were mainly distributed in frontal-cortex, lateral septal nucleus, basolateral amygdaloid nucleus, hippocampus CA1, paraventricular nucleus, ventral posterolateral nucleus, habenular nucleus, ventromedial hypothalamus nucleus, supraoptic nucleus, substantia nigra nucleus, interpeduncular nucleus, periaqueductal gray and dorsal horn. In enflurane group the number of FLI and ELI neurons in these nuclei was significantly higher(P
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The experiment of recirculating blood perfusion device showed that trimetazidine (TMZ ) 30 mol/L could significantly inhibit the arrhythmias induced by ischemia and reperfusion, decrease the content of malondialdehyde and improve superoxide dismutase activity of red blood cell in isolated rabbit hearts. In the experiment with coronary artery occlusion of rats, TMZ 4mg/kg administered intravenously could also inhibit arrhythmias induced by ischemia and reperfusion. The mechanism may be related to the depression of lipid peroxidation reaction.
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Objective To determine the effects of enflurane and isoflurane on the spontaneous neural discharge of central amygdaloid nucleus in rats Methods SD rats (30 60 d) of either sex were decapitated Brain was immediately removed and kept in 4℃ artificial cerebral spinal fluid(ACSF) which was balanced with 95% O 2 and 5% CO 2 gas mixture Braine tissue containing central amyfdaloid nucleus was cut into slices of 300 400?m thick which were immersed in ACSF Enflurane and isoflurane were administered by balancing ACSF with enflurane (1 5%,3 0%,4 5%) or isoflurane (1 1%,2 2%,3 3%) The spontaneous neural discharge of central amygdaloid nucleus was measured before and after enflurane or isoflurane using whole cell patch clamp techniques Results Enflurane and isoflurane inhibited the frequencies of spontaneous neural discharge of central amygdaloid nucleus in a dose dependent manner The spontaneous neural discharge inhibited by enflurane (3 0%) and isoflurane (2 2%) could recover after the slices being washed with normal ACSF for 5 min Conclusions The results indicate that the spontaneous neural discharge of central amygdaloid nucleus can be inhibited reversibly by enflurane and isoflurane Central amygdaloid nucleus may by one of the sites of action of enflurane and isoflurane in central nervous system
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Objective To investigate whether spinal cord is involved in the analgesic effect of propofol.Methods Fifteen adult SD rats of both sexes weighing 195～223g were randomly divided into three group of five animals each: control group received normal saline intraperitoneally(ip), fentanyl group received fentanyl 0 1mg/kg and propofol group propofol 100mg/kg ip 2 min later 4% formalin 150?l was injected subcutaneously into the planta region of right hindpaw 1h after formalin injection animals of all three groups were anesthetized with pentobarbital sodium ip After induction of anesthesia chest was open and 100 ml of normal saline was infused via left ventricle then followed by 4% formalin infusion for fixation of tissue, 90 min later spinal cord, L 3 5 sect, was removed for determination of c fos expression in spinal cord using fos immunohistochemistry technique Results In control group in less than 10 s after formalin intraplantar injection the animals were agitated, restless, lame and paw licking In fentanyl group and propofol group the righting reflex was suppressed for (19 4?7 8) min and (7 2?1 5)min and no pain response was seen during this period When righting reflex recovered the pain response was much lighter than that in the control group Formalin stimulation induced c fos expression was seen only in the ipsilateral spinal cord Both fentanyl and propofol significantly suppressed c fos expression evoked by formalin stimulation The number of fos like immunoreactivity neurons(FLIN) decreased by 57 8% and 36 3% respectively(P
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Objective To investigate the effects of inhalation of different concentrations of isoflurane on global and regional cerebral glucose metabolic rate (CMRglu) in healthy volunteers using positron emission tomography (PET) scan.Methods The study was approved by the ethic committee of the hospital. After obtaining written informed consent we studied eight right-handed healthy volunteers (3 male, 5 female), aged 21-28 yrs. Each volunteer underwent 3 PET scans.They were fasted for 8 h prior to study. The PET scan was performed when conscious, at 0.5 and 1.0 MAC isoflurane anesthesia. The interval between two PET scans was 1 week. Scans were obtained with a MASEP CPET Plus scanner (2.0 mm resolution-FWHM) using the 18 fluorodeoxyglucose technique. Results The whole brain CMRglu averaged 30.0?1.1 ?mol?100 mg-1?min-1 when the volunteer was awake. Isoflurane anesthesia significantly reduced whole-brain CMRglu by 24% to 23.3?1.4 ?mol?100 mg-1?min-1 at 0.5 MAC and by 41% to 18.4?0.9 ?mol?100mg-1?min-1 at 1.0 MAC (P
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0.05) in VAS and ramsay scale, but side effects were significantly different in group B compared with those in group S. Conclusion: The analgesia efficacy of S is same as B in maxillofacial postoperation, but side effects are more significantly in group B, and group S is more safe and handy.
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Objective:To observe the sedation effects of midazolam on maxillofacial postoperative patients with nasotracheal intubation. Methods:40 cases were equally and randomly divided into two groups: sufentanil group (group S) and sufentanil-midazolam group(group S-M).The continuous dose was 2 ml/h. If the patients feel uncomfortable, patient controlled intravenous analgesia (PCIA) was used. To observe the changes of circulation and respiration,the changes of visual analogue scale(VAS) and Ramsay score, the patients' bucking times within 24 h and the PCIA times, patients' total satisfactory degree and the complications during postoperative analgesia were recorded. Results:There were significant decreases in VAS of two groups after postoperative analgesia 1, 4, 12, 24, 48 hours(P
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Objective To investigate whether oral propofol has any inhibitory effects and dose-response relationship on the pain inducing tissue injury in mice. Methods The effect of propofol on pain was observed in formalin test and acetic acid writhing test in mice. Formalin was injected subcutaneously into the plantar surface of one hind paw. Spontaneous nocuous responses were immediately scored by counting the number of flinches of the injected hindpaw at every 5-minute interval during a 60-minute period. The number of writhing caused by intraperitoneal injection of 0.6% acetic acid was also observed in mice. Results Oral propofol in a dose of 100mg/kg did not significantly inhibit nocuous stimulation. With higher doses, propofol inhibited both the phases 1 and 2 of persistent spontaneous pain induced by subcutaneous formalin injection. Orally taken propofol also inhibited the number of writhing after intraperitoneal acetic acid injection in a dose dependent manner. Conclusion Oral propofol is effective in inhibiting pain induced by formalin and acetic acid.
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Objective To study the protective effect of a hyperoxic solution on phosgene-induced lung injury by observing the changes in W/D ratio, lung water (LW), and L/B, and MDA contents, GSH-PX activity, and protein contents in broncho-alveolar lavage fluid (BALF). Methods The rabbits were divided into normal control group, hyperoxic solution (HO) and balance salt(BS) groups.Group HO and Group BS inhaled phosgene, and hyperoxic solution was given intravenously in group HO, but BS was given in group BS. W/D, LW, L/B, and MDA contents,GSH-PX activity,protein contents in broncho-alveolar lavage fluid (BALF) were determined. Results The MDA contents, W/D, LW and L/B were increased, and GSH-PX activity was decreased significantly in Group HO and Group BS compared with control group (P
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Objective To investigate the effects of inhaling isoflurane and enflurane on the spontaneous neural discharge of the neurones in rat hippocampus. Methods Whole cell patchclamp recording te chnique was used to observe the effects of isoflurane and enflurane on the spont aneous discharge rate of the neurons in the hippocampus on the brain slice of ne w-born SD rats. After decapitation, the whole brain of the rat was removed and put into artificial cerebrospinal fluid (ACSF) saturated with 1.36 g?L -1 O 2 and 0.098 g?L -1 CO 2 mixed gas at 4 ℃ . Brain was cut into 300～400 ? m thick slices containing the hippocampus. Whole cell patchc lamp recording technique was used to observe the effects of isoflurane with dif ferent concentrations on the spontaneous discharge rate of neurons in the hippoc ampus on the brain slices. Results Isoflurane and enfluran e could significantly inhibit the spontaneous neural discharge of neurons in the hippocampus in a dose-dependant manner. The effects of spontaneous neural disc harge of hippocampus inhibited by isoflurane (0.12 g?L -1 ～0.36 g?L -1 ) and enflurane (0.2 g?L -1 ～0.6 g?L -1 ) could be recove red following washing off with ACSF for 5 min. Conclusion T he spontaneous discharge rate of neurons in the hippocampus can be reversibly in hibited by isoflurane and enflurane. Hippocampus may be an important action site of anesthetics isoflurane and enflurane in the central nervous system.
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Objective: To evaluate the effect of propofol and penthiobarbital on the postoperation conscious level in patients after general anesthesia. Methods: 116 patients with maxillofacial operation were randomly divided into two groups. Propofol was used in 60 cases and penthiobarbital in 56 for sedation. Wake score,anoxia and local stimulation in vein were examed.Results: In the 60 cases treated with propofol anoxia and local vein stimulation were observed in 2 and 7 cases, the restlessness score and the consciousness scores were 1.28?0.76 and 1.9?0.7; in the 56 cases treated with penthiobarital those were in 13 and 35 (P
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<p><b>OBJECTIVE</b>To study the effects of cardiovascular function in dental extraction on hypertensive patients by inhalation 50% nitrous oxide.</p><p><b>METHODS</b>The 30 hypertensive patients were randomly allocated into two groups: A group inhalation the 50% nitrous oxide and oxygen, B group inhalation the air and O(2). To measured the HR, BP, and SpO(2) in dental extraction.</p><p><b>RESULTS</b>In a group the changes of blood pressure and heart rate are more smoother than B group. Two groups were significant in HR, BP and SpO(2) (P < 0.01).</p><p><b>CONCLUSIONS</b>Inhalation with 50% nitrous oxide can keep the stability of cardiovascular system and it is a valid method in dental extraction on hypertensive patients.</p>
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Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Administração por Inalação , Anestésicos Inalatórios , Farmacologia , Pressão Sanguínea , Frequência Cardíaca , Hemodinâmica , Hipertensão , Óxido Nitroso , Farmacologia , Oxigênio , Sangue , Pressão Parcial , Fatores de Tempo , Extração DentáriaRESUMO
AIM: To evaluate the protective effect of rapid phase of ischemic preconditioning against spinal cord ischemic injury in rabbits. METHODS: Thirty six male New Zealands white rabbits were randomly assigned to 3 groups (12 in each group): ischemia and reperfusion injury group (IR group), ischemic preconditioning + IR group (IPC+IR group) and sham operation group (sham). In IR group, spinal cord ischemia was induced by an infrarenal aorta clamping for 20 min; The rabbits in IPC+IR group underwent a 6 min ischemic preconditioning followed by 30 min of reperfusion before the 20 min clamping; The rabbits in sham group underwent the same procedures as the IR group except for infrarental aortic unclamping. Neurologic status was scored at 8, 12, 24 and 48 h after reperfusion. All animals were sacrificed at 48 h after reperfusion and the spinal cords (L_(5-7)) were removed for histopathologic study and determination of the activity of Na~+, K~+-ATPase. RESULTS: The neurologic function scores in sham group and IPC+IR group at each observation interval were higher than those in IR group (P