Objective To study the effect of remifentanil precondition on myocardium against ischemia/ reperfusion (I/R) injury. Methods Fifty male SD rats weighing 300-350 g were anesthetized with intraperitoneal 5% pentobarbital 40 mg?kg-1, tracheostomized and mechanically ventilated. Right carotid artery and jugular vein were cannulated for BP monitoring and drug administration. ECG, BP and HR were monitored continuously. The chest was opened and heart exposed via a left thoracotomy. A 6-0 prolene suture was placed around the left coronary artery and a snare was made. The coronary artery was occluded by tightening the snare. Myocardial ischemia was confirmed by the appearance of a regional cyanosis of myocardium, decrease in BP and ST-T changes on ECG. After the surgical procedure a 15-minute stabilization period was allowed. The animals were then randomly divided into 3 groups: Ⅰ control group (CON, n = 9) ; Ⅱ ischemic preconditioning group (IPC, n = 9) and Ⅲ remifentanil preconditioning group which was further divided into 5 subgroups according to the dose of remifentanil (RPC) . In control group the hearts were subjected to 30 min ischemia followed by 120 min reperfusion (I/R). In IPC group the hearts were subjected to 3 episodes of 5 min ischemia in succession at 5 min interval for reperfusion before I/R. In RPC group the animals received 3 periods of 5 min remifentanil infusion at 0.2 (RPC1 , n = 5), 0.6 (RPC2,n = 6), 2 (RPC3 ,n = 8), 6 (RPC4 ,n = 7) or 20 (RPC5 ,n = 6) ?g?kg-1?min-1 at 5 min interval before I/R. MAP, HR and RPP (SP?HR) were recorded before and after I/R. Infarct size (IS) and the area at risk (AAR) were determined by triphenyltetrazolium staining. Results MAP was significantly lower at the end of 30 min ischemia in RPC2, RPC3, RPC4 and RPC5 subgroups than in control group. There was no significant difference in MAP, HR and RPP at the end of 120 min reperfusion among the 3 groups. The infarct size was significantly smaller in IPC and RPC groups than in control group. RPC at 6 ?g?kg-1?min-1 provided best protective effect. The sigmoidal equation of the dose-effect curve was Y = 15.18 + 17.76/ [1 + 10(-2.57-X)]. ED50, was 2.689 ?g?kg-1?min-1.Conclusion Remifentanil and IPC have similar protective effect on the heart against I/R injury.

OBJECTIVE To investigate the protective effect of urantide on myocardial apoptosis in rats induced by ischemia/reperfusion (I/R) or hypoxia/reoxygenation (H/R) injury and explore the underlying mechanism. METHODS ① In vivo test A rat myocardial I/R injury model was induced by ligating and untying the left anterior descending coronary artery with occlusion 30 min/reperfusion 60 min. Urantide 3, 10 and 30 μg·kg-1 was iv given 10 min before ischemia. TUNEL labeling was used for apoptosis measurement in myocardium. Immu-nohistochemical assay was used for Bcl-2 and Bax proteins expression detection. ② In vitro test An H/R cell model was set up by 3 h hypoxia/3 h reoxygenation. Urantide 0.1,1 and 10 nmol·L-1 was added just before hy-poxia, respectively. Hoechst33258 assay and flow cytometric techniques were used to detect apoptotic cells. RESULTS ① In vivo test Compared with sham group, the number of TUNEL-positive cells in I/R model group significantly increased (P<0.01) ; Bcl-2 protein expression slightly increased with no significant difference, Bax protein expression markedly increased ( P < 0. 01 ) , while Bcl-2/Bax ratio in I/R model group significantly decreased (P <0.01). Compared with I/R model group, the number of TUNEL-positive cells in urantide 10 and 30 μg·kg-1 groups was significantly decreased by about 36.6% and 57. 2% (P<0.05) ; Bax protein expression markedly decreased ( P <0.05 ) , while Bcl-2/Bax ratio was significantly augmented ( P <0.05 ). Urantide 30 u,g-kg1 also markedly increased Bcl-2 protein expression(P <0.05). ② In vitro test Compared with normal control group, the apoptosis rate in H/R model group significantly increased (P<0. 01). Hoechst33258 assay revealed that urantide 0.1, 1 and 10 nmol·L-1 reduced H/R-induced apoptotic nuclei by about 27.9% , 59.0% and 75. 4% , respectively (P <0.05). Flow cytometric techniques showed that the apoptosis rate was significantly reduced by about 32.8% and 64. 7% with administration of urantide 1 and 10 nmol·L-1 (P < 0. 01). CONCLUSION Urantide exerts an inhibitory effect on I/R or H/R-induced apoptosis by increasing Bcl-2 protein expression and decreasing Bax protein expression.

Aim To investigate the protective effect of the potent UT receptor(urotensin Ⅱ receptor,UTS2R)antagonist-urantide against myocardial ischemia-reperfusion(I/R)injury and its probable mechanisms in rats.Methods Rat myocardial I/R injury was induced by ligating and untying the left anterior descending coronary artery.The rats were randomly assigned into 6 groups:sham group,model group,urantide 3 ?g?kg-1 group,urantide 10 ?g?kg-1 group,urantide 30 ?g?kg-1 group and Ver(Verapamil)1.6 mg?kg-1 group.All animals except sham group were subjected to 30 min of occlusion and 60 min of reperfusion.Urantide or Ver was given ten minutes before occlusion through intravenous drug perfusion.Heart rate(HR)and the ST segment change of electrocardiogram(ECG)were recorded.The content of malondialdehyde(MDA)and nitric oxide(NO),and the activity of lactate dehydrogenase(LDH)and nitric oxide synthase(NOS)in blood serum were measured.Infarct size(IS),as a percentage of the area at risk(AAR),was determined by Evans blue and TTC double staining.The expression of iNOS protein was detected by western blotting.Results The results demonstrated that during the process of I/R,HR decreased significantly whereas ST segment of ECG markedly elevated.After I/R,MDA content and LDH activity in blood serum increased significantly,while total NO content and total NOS activity decreased sharply.Urantide(10,30 ?g?kg-1)had no significant effect on HR changes,but could markedly inhibit the elevation of ST segment of ECG,MDA content and LDH activity,and inhibit the decline of total NO content and total NOS activity,at the same time decrease I/R induced IS/AAR.But 3 ?g?kg-1 urantide had no significant effect on above indexes.Except for this,urantide(10,30 ?g?kg-1)down-regulated the I/R induced expression of iNOS.Conclusions Our findings indicate that urantide has a protective effect against myocardial I/R injury in rats.The cardio-protective involves the inhibition of lipid peroxidation and the stimulation of NO release.

Aim To study the protective effect of total flavones of rhododendra pharmacological preconditioning(TFR-PP)on myocardial ischemia and reperfusion injury and its mechanisms involved in myocardial inflammation in rats.Methods In model group the isolated perfused rat hearts set up by Langendorff system were subjected to 30 min ischemia followed by 40 min reperfusion. The hearts in TFR-PP groups were subjected to three cycles of 5 min perfusion with and without TFR before 30 min ischemia followed by 40 min reperfusion. In all groups the activities of lactate dehydrogenase (LDH), creatine phosphokinase (CK), and myeloperoxidase (MPO) in myocardium, the expressions of nuclear factor-kappa B (NF-?B), tumor necrosis factor-alpha(TNF-?), and intercellular adhersion molecule-1 (ICAM-1) were measured, and the myocardial pathomorphological changes were examined.Results TFR-PP(25、50、100 mg?L-1) could inhibit markedly the reductions of LDH and SOD activities in myocardium induced by ischemia and reperfusion injury.100 mg?L-1 TFR-PP also significantly improved the pathomorphological changes of injury. TFR-PP (25、50、100 mg?L-1) could inhibit the expressions of NF-?B, TNF-? and ICAM-1 to varying degrees.Conclusion TFR-PP has marked protective effect on ischemia and reperfusion injury in isolated rat heart via inhibiting the inflammation of myocardium.

Objective To quantify the percentage of CD8+ T cells and their expressions of cytotoxic molecules and homing-related chemokine receptors in peripheral blood from patients with atopic dermatitis (AD).Methods Peripheral blood was obtained from 15 patients with AD and 14 healthy controls.Flow cytometric analysis was performed to determine the percentages of CD8+ T cells and CD8+CLA+ T cells in the peripheral blood samples,as well as the expression levels of cytotoxic molecules and homing-related chemokine receptors on these cells.Differences in these parameters were analyzed using t test,and relationship between these parameters was evaluated using Pearson correlation coefficient.Results No significant difference was observed between the patients with AD and healthy controls in the percentage of CD8+ T cells,the expressions of perforin,granzyme B,CCR10,CCR6 or FasL on CD8+ T cells,or the expressions of CCR4,CCR10,CXCR6 or FasL on CLA+CD8+ T cells (all P ＞ 0.05).A significant increase was noted in the percentage of CLA+CD8+ T cells (3.80％ ± 1.46％ vs.2.18％ ± 0.85％,t =3.636,P ＜ 0.01) and expression rates of CCR4 on CD8+ T cells (13.86％ ± 4.42％ vs.9.50％ ± 2.14％,t =3.738,P ＜ 0.01) as well as perforin and granzyme B on CLA+CD8+ T cells (74.27％ ± 15.94％ vs.57.20％ ± 14.64％,t =2.998,P ＜ 0.01; 70.90％ ± 13.85％ vs.56.41％ ± 11.00％,t =3.104,P ＜ 0.01) in the patients with AD compared with the healthy controls.Conclusions The proportion of CLA+CD8+ T cells is increased with enhanced expressions of cytotoxic molecules such as perforin and granzyme B in peripheral blood of patients with AD,which may contribute to the pathogenesis of AD.

Aim To determine whether remifentanil preconditioning has cardioprotection against ischemia and reperfusion induced injury via opioid receptor(OR) in rats.Method Male Sprague-Dawley rats of 300～350 g were anaesthetized and the chests were opened and hearts exposed via a left thoracotomy.They were randomly assigned to 12 groups:Control(CON,saline vehicle),naltrindole(NTD,5 mg?kg~(-1) iv.10 min before 30 min ischemia);CTOP(CTOP,1 mg?kg~(-1) iv.10 min before ischemia),nor-binaltorphimine(nor-BNI,5 mg?kg~(-1),iv.15 min before 30 min ischemia);remifentanil preconditioning(RPC,6 ?g?kg~(-1)?min~(-1)),ischemic preconditioning(IPC),NTD+RPC or NTD+IPC(naltrindole 5 mg?kg~(-1) iv.10 min before RPC or IPC),nor-BNI+RPC or nor-BNI+IPC(nor-binaltorphimine 5 mg?kg~(-1),iv.15 min before RPC or IPC),CTOP+RPC or CTOP+IPC(CTOP 1 mg?kg~(-1) iv.before RPC or IPC).Infarct size(IS),a percentage of the area at risk(AAR),was determined by triphenyltetrazolium(TTC) staining.Results IPC and RPC markedly reduced IS/AAR from(52.7?1.8)% to(12.9?2.7)% and(16.2?2.4)%,respectively. CTOP,a selective ?-OR antagonist,or NTD,a selective ?-OR antagonist,administered 10 min before remifentanil PC completely abolished,while nor-BNI,a selective ?-OR antagonist,administered 15 min before RPC attenuated the cardioprotective effect of RPC.In the group preconditioned with ischemia,blockade of ?-OR with NTD abolished,while blockade of ?-OR with nor-BNI attenuated the protection.Blockade of ?-OR with CTOP did not alter the cardioprotective effect of IPC.Conclusion The cardioprotective effect of RPC was abolished by all the three OR antagonist,CTOP,naltrindole and nor-binaltorphimine,suggesting that this effect is mediated via ?-,?-and ?-ORs.Part of the effect of RPC may be produced by ?-OR agonist activity outside the heart.

Objective To evaluate the role of δ receptor in reduction of hypoxia-reoxygenation (H/R) injury to cardiomyocytes by morphine preconditioning in rats with chronic heart failure.Methods Adult male Sprauge-Dawley rats weighing 220-250 g were used in the study.Chronic heart failure was induced by injection of adriamycin 2 mg/kg via the tail vein once a week for 6 weeks.Their hearts were removed 2 weeks after the last injection and the cardiomyocytes were isolated and cultured.The cells were randomly divided into 5 groups ( n =8 each):control group (group C); H/R group; morphine preconditioning group (group MPC); morphine preconditioning + naloxone (opioid receptor antagonist ) group (group MPC + Naloxone) ; morphine preconditioning + naltrindole ( δ receptor antagonist) group ( MPC + Naltrindole group).The cells were cultured in normal culture atmosphere in group C and were exposed in hypoxic air for 3 h followed by 1 h reoxygenation in the other groups.Morphine preconditioning was performed immediately before hypoxia in group MPC.Naloxone and naltrindole were added before morphine preconditioning in groups MPC + Naloxone and MPC + Natrindole respectively.At 1 h of reoxygenation,the cell viability ( by MTT assay),activities of lactate dehydrogenase ( LDH ) and creatine kinase (CK),and cell apoptosis were detected.The apoptotic rate was calculated.Results The cell viability was significantly lower,and the activities of LDH and CK and apoptotic rate were significantly higher in groups H/R,MPC + Naloxone and MPC + Natrindole than in group C (P ＜ 0.05).The cell viability was significantly higher,and the activities of LDH and CK and apoptotic rate were significantly lower in group MPC than in group H/R ( P ＜ 0.05).Conclusion Morphine preconditioning reduces H/R injury to cardiomyocytes through activating δ receptor in rats with chronic heart failure.

Objective:This study was aimed to investigate the value of osteoclast differentiation factor (ODF) and osteoclastogen-esis inhibitory factor (OCIF) detection for clinical diagnosis and assessment of patient condition in bone metastasis of lung cancer. Methods:Data from 186 lung cancer patients who were preliminary diagnosed between July 2009 and April 2012 were analyzed. Cas-es were divided into the bone metastasis group with 82 cases (group A) and the non-bone metastasis group with 104 cases (group B). Concentrations of serum ODF and OCIF in each group were detected by ELISA. Results: ODF and OCIF values of group A were (32.22±6.22) ng/L and (41.23±8.13) ng/L, respectively, which were significantly higher than the corresponding values in group B [(8.35 ±5.42) ng/L and (10.15±4.42) ng/L]. The differences between the two groups were statistically significant (P<0.01). Areas under the re-ceiver operating characteristic curves of ODF and OCIF, which are used to diagnose bone metastasis in lung cancer, were 0.91 and 0.87, respectively, manifesting good diagnostic value. The sensitivity and specificity of ODF in diagnosing lung cancer with bone metastasis were 90.38%and 86.59%, respectively, and those of OCIF were 86.54%and 84.15%, respectively. ODF increased, whereas OCIF de-creased significantly, with increasing bone metastasis. ODF and OCIF concentrations in group A and the group with newly-found bone metastasis were significantly higher than those in group B, with statistically significant differences among these groups (P<0.01). Com-pared with group A, less difference was found in the ODF and OCIF of newly-found bone metastases, without statistical significance be-tween these groups (P>0.05). Conclusion:The serum ODF and OCIF concentrations significantly increase when bone metastasis oc-curs in lung cancer patients. Hence, these variables are useful as indices for monitoring bone metastases and evaluating patient condi-tion. An extensive application prospect is proposed.

Objective To study the protective effect of total flavone from Rhododendron simsii(TFRS) on myocardial ischemia-reperfusion injury rats and its mechanism.Methods The ischemic model was made by occluding the anterior descending of the left artery(LAD) in rats.The change of ST segment and T wave of electrocardiograph(ECG) were observed,and the activity of lactate dehydrogenase(LDH),creatine kinase(CK),levels of the maleic dialdehyde(MDA),and nitric oxide(NO) in serum were measured.And by tetrazolium chloride(TTC) staining,the areas of myocardial infarction were observed.The expression of inducible nitricoxide synthase(iNOS) in rats was detected by emploring the reverse transcription-polymerase chain reaction(RT-PCR) technique.Results On the myocardial infarction model by occluding the anterior descending of the LAD in rats,TFRS(100 mg/kg) obviously reduced the height of ST segment after occluding 30 min and TFRS(25,50,and 100 mg/kg) obviously reduced the height of ST segment after reperfusion 30 min.TFRS(50 mg/kg) reduced by myocardial infarction area.TFRS(50 and 100 mg/kg) obviously reduced the activity of CK and LDH.TFRS(50 mg/kg) decreased the level of MDA in serum.By RT-PCR technique,it was found that the expression of iNOS mRNA in myocardium in IR rats pretreated with TFRS(100 mg/kg) was higher than that in IR and normal groups.Conclusions TFRS has the significant protection against myocardial ischemia-reperfusion injury via atte-nuating oxygen free radical and increasing the expression of iNOS mRNA and NO production.

Aim To investigate the effect of intracerebroventricular morphine postconditioning against ischemia-reperfusion injury in rat heart and the mechanism of the central nervous system opioid receptor.Methods Forty-two Sprague-Dawley Rats were established intracerebroventricular catheter placement and myocardial ischemia/reperfusion models and randomly assigned to 7 groups:Sham group(Sham),control group(CON),intravenous control group(VCON),morphine postconditioning group(POC),intracereborventricular morphine postconditioning group(MOC).According to the dosage of intracerebroventricular morphine(3 ?g?kg-1,0.3 ?g?kg-1,0.03 ?g?kg-1),MOC group was assigned to three groups :MOC 1,MOC 2,MOC 3.Infarct size(IS),a percentage of the area at risk(AAR) was determined by triphenyltetrazolium(TTC) staining.c-fos expression in nucleus of tractus solitarius was determined by immunohistochemical method and Cardiac TroponinI(cTnI) of serum was observed at 120 min of reperfusion.Results Compared with control group,IS,IS/AAR and cTnI were significantly reduced in POC and MOC groups(P0.05).c-fos expression in nucleus of tractus solitarius were significantly reduced in MOC 2,POC(P