Objective To investigate the effects of distraction on tourniquet pain in healthy subjects .Methods Tourniquet pain was induced by the tourniquet at continuous pressure of 200 mm Hg for 10 min .20 healthy college students were asked to perform two distraction tasks during the pressure ,one of which is pictorial dot-probe task with three kinds of emotional pictures ,the other is word dot-probe task with five kinds of pain-related words ,and a control task ,in which no distraction task was performed .The pain intensity ,pain distress were recorded by Visual Rating Scale(VRS)and Modified McGill Pain Questionnaire short-form(MPQ-SF) . Results Compared with the control task[(4 .1 ± 1 .8) ,(4 .0 ± 1 .8)] ,the pain intensity and distress were significant lower in picture [(3 .1 ± 1 .3) ,(3 .0 ± 1 .2)]and word[(3 .3 ± 1 .4) ,(3 .4 ± 1 .5)] distraction tasks[(F(2 ,8)=21 .424 ,P<0 .001;F(2 ,8)=17 .962 ,P<0 .001)] .The pain distress in word distraction task (3 .4 ± 1 .5) was higher than that in the pictorial distraction task (3 .0 ± 1 .2) (P<0 .05) .Meanwhile ,for the last four minutes ,the pain intensity was significant lower in pictorial distraction task compared with the control task(P<0 .05) .And at the beginning of the experiment ,the pain distress was significant lower in pictorial[(2 .3 ± 0 .7) , (2 .5 ± 0 .8)]and word[(2 .4 ± 0 .8) ,(2 .9 ± 0 .9)] distraction task compared with the control task [(3 .7 ± 1 .3) ,(4 .0 ± 1 .4)](P<0 .05) .Compared with the control task (5 .0 ± 1 .6) ,the present pain index (PPI) was significant lower in the pictorial distraction task(3 .5 ± 1 .4)(F=5 .097 ,P<0 .05) .Conclusion The tourniquet pain was attenuated by distraction of cognitive tasks in pain in-tensity and pain distress .

Objective To explore the role of P38 mitogen-activated protein kinase (P38 MAPK) in the development of cerebral hypoxic preconditioning. Methods Healthy male BALB/C mice weighted as18～20 g were randomly divided into 7 groups as follows: normoxic control (H0), early (H1～H4) and delayed (H5 and H6) hypoxic preconditioned mice groups. SDS-PAGE, Western blot and Gel Doc imagine systems were applied to quantitatively analyze the level of P38 MAPK phosphorylation and protein expression in the brain of mice. Results The phosphorylation levels of P38 MAPK increased in cortex, hippocampus and hypothalamus of mice in both early (H1～H4) and delayed (H5 and H6) hypoxic preconditioned groups, and the statistic significance (P

Objective To investigate the protective effects of Bauhinia championii Benth flavones(BCF) on oxidative stress of neonatal rat cadiocytes induced by H2O2.Methods Cadiocytes of neonate rat was cultivated for 72 hours and divided into six groups: a normal control group, a H2O2 group, BCF(60, 120 and 240μg/ml)+H2O2 groups and aShuxueninginjection (100μg/ml)+H2O2 group (n=8). 6 hours after the drugs were given, the morphology changes was observed and the survival rate was detected; the content of AST, CPK, LDH in culture medium were detected; the activity of SOD, CAT, GSH-Px and the content of MDA in cardiomyocytes were also determinted; the apoptosis rate were detected, and the expression of caspase-3 in cardiomyocytes was detected by Western blot.Results Compared with the H2O2 group, the activity of AST(28.8 ± 6.1 U/ml, 24.5 ± 5.3 U/mlvs. 36.2 ± 6.7 U/ml), CPK(1.8 ± 0.4 U/ml, 1.5 ± 0.3 U/mlvs. 2.5 ±0.4 U/ml), LDH(805.2 ± 160.9 U/L, 671.5 ± 128.7 U/Lvs. 916.5 ± 168.4 U/L) in culture medium were significantly decreased (P<0.05,P<0.01), the activity of SOD(84.8 ± 17.4 U/mg, 95.3 ± 18.2 U/mgvs. 55.7 ± 13.1 U/mg), CAT(23.4 ± 3.1 U/mg, 26.3 ± 3.5U/mgvs. 15.2 ± 3.0 U/mg) in cardiomyocytes were significantly increased and the content of MDA(8.1 ± 1.7 nmol/mg, 6.8 ± 1.5 nmol/mgvs. 11.1 ± 2.3 nmol/mg) were significantly decreased (P<0.05,P<0.01), the expression of c caspase-3(1.64 ± 0.16, 1.30 ± 0.12vs. 2.06 ± 0.25) and the apoptosis rate (24.2% ± 5.5%, 13.4% ± 3.9%vs. 51.2% ± 9.1%) were significantly decreased (P<0.05,P<0.01); the activity of GSH-Px (3.6 ± 0.9 U/mg vs. 2.4 ± 0.7 U/mg) in cardiomyocytes of BCF 240μg/ml treatment group was increased (P<0.05).Conclusions BCF could effectively improve the morphology of neonatal rat cadiocytes induced by H2O2, increase the survival rate, improve the activity of antioxidase, down-regulate the expression of caspase-3 and decrease the apoptosis rate, suggesting that BCF had dose-dependent protective effects on oxidative stress of neonatal rat cadiocytes induced by H2O2.

Objective To explore the role of conventional protein kinase C(cPKCs) in delayed hypoxic preconditioning.Methods The biochemistry techniques of SDS-PAGE,Western bolt and Gel Doc imagine were applied to analyze the effect of repetitive hypoxic exposure(H5,H6) on the level of cPKC?,? membrane translocation and protein expression in murine brain.Results We found that cPKC? protein expression was significantly decreased(P

Objective To determine the changes in phosphorylation of conventional protein kinase C?(cPKC?)-interacting extracellular signal regulated kinase 1/2(ERK1/2) in the hippocampus of hypoxic preconditioned(HPC) mice.Methods Healthy male BalB/c mice were used to develop "auto-hypoxia"-induced HPC mice and randomly divided into 3 groups as follows: normoxic control(H0),early(H3) and delayed hypoxic preconditioned groups(H6).Co-immunoprecipitation and Western blot were applied to quantitatively analyze the phosphorylation level of cPKC?-interacting ERK1/2 in cytosolic and particulate fractions of hippocampus of HPC mice.ResultsERK1/2 was co-immunoprecipitated by cPKC? in hippocampus of mice.The phosphorylation level of cPKC? interacting ERK1/2 at tyrosine 204 decreased significantly in the hippocampus of H3 and H6 groups as compared with that of the H0 group(P

Objective To evaluate the effect of delayed preconditioning with morphine on ischemic cerebral injury in mice and the role of classical protein kinase C (cPKC).Methods Forty male BALB/C mice,weighing 20-22 g,were randomly divided into 4 groups (n =10 each):sham operation group (group S),ischemic cerebral injury group (group ICI),morphine preconditioning group (group MP) and cPKC inhibitor Go6983 group (group G).Ischemia was induced by middle cerebral artery occlusion (MCAO).In S group,the middle cerebralartery was only exposed but not occluded.In MP group,morphine 10 mg/kg was injected intraperitoneally 24 h before MCAO.In G group,morphine 10 mg/kg was injected intraperitoneally 24 h before MCAO and 5 μl Go6983 (6nmol) was injected into the left lateral cerebral ventricle immediately before MCAO.The neurologic deficit was evaluated and scored according to neurological disability status scale in a blind nanner 6 h after MCAO.The animals were sacrificed and brains were immediately removed for measurement of the brain edema and infarct volume.Apoptotic rate was calculated.Results Compared with S group,the neurologic deficit scores,infarct volume,brain edema and apoptotic rate were significantly increased in ICI,MP and G groups (P ＜ 0.01).Compared with group ICI,the neurologic deficit scores,infarct volume,brain edema and apoptotic rate were significantly decreased in group MP (P ＜ 0.01),and no significant change was found in the parameters mentioned above in group G (P ＞ 0.05).Conclusion Delayed preconditioning with morphine can reduce ischemic cerebral injury in mice and activation of classical cPKC signaling pathway is involved in the mechanism.

Objective To investigate the factors affecting the protective effects of morphine preconditioning on murine hippocampal neurons against anoxia/reoxygenation (A/R) injury and the underlying mechanisms.Methods Hippocampal slices (400 μm thick) were prepared using hippocampi isolated from decapitated mice. A/R injury was simulated in vitro using artificial cerebral spinal fluid (ACSF) deprived of O_2 and glucose for 20 min followed by reoxygenation and glucose supply for 2 h. The experiment was performed in 4 parts: I .The slices were incubated with 5 different concentrations of morphine (0.1, 0.3, 0.5, 1.0, 3.0, 10.0 /μmol/L) for 30 min at 30 min before A/R; Ⅱ. The slices were incubated with morphine 3.0 /μmol/L for 5 different periods of time (5, 15, 30, 45, 60 min) at 30 min before A/R; Ⅲ. The slices were incubated with morphine 3.0 μmol/L for 30 min followed by A/R at 6 different intervals (0, 5, 15,30,60, 120 min); Ⅳ. The slices were incubated with (a) chelerythrine (a non-selective PKC antagonist) 10 /μmol/L or (b) εVl-2 (a selective nPKCe isoform antagonist) 2 μmol/L or (c) AIP 2 μmol/L (a selective CaMK Ⅱ antagonist) or (d) MK-801 10 μmol/L (a non-competitive NMDA receptor blocker) for 30 min and then for another 30 min together with morphine 3.0 μmol/L before A/R at 30 min interval. The survival rates of the hippocampal neurons were assessed by TTC staining. Results Neuronal survival rates were significantly higher in morphine preconditioning groups which preconditioned with morphine (0.5-10.0 μmol/L) for 15-60 min at an interval of 0-60 min before A/R than in A/R group. Increase in neuronal survival rate induced by morphine preconditioning was partially blocked by chelerythrine or εV1-2 or AIP or MK-801. Conclusion Preconditioning with appropriate concentrations of morphine (0.5-10.0 μmol/L) for appropriate period of time (15-60 min) at appropriate interval (within 60 min) before A/R can protect hippocampal neurons against A/R injury through activation of nPKCε, NMDA receptor and CaMKⅡ.

Objective To identify certain isoforms involved in the onset of retinal ischemic preconditioning (IPC), the effects of ischemic pretreatment (IP) were observed on level of conventional protein kinase C (cPKC) ?,?Ⅰ、?Ⅱand ? isoform-specific membrane translocation and protein expression in retina of rats. Methods Retinal IP was produced by intra-ocular pressure (IOP) elevation for 5 minutes in anesthetized Wistar rats. Sham operation was similar to IP except the pressure elevation. 10, 20, or 40 minutes and 1, 12, 24, 72 or 168 hours after the procedure, cPKC isoform-specific membrane translocation and protein expression were analyzed using Western-blot. Results cPKC? protein expression significantly increased from 12 h to 168 h. A peak reached at 72 h after IP. cPKC? membrane translocation enhanced during 20 min to 1 hours with a peak at 40 min. cPKC? protein expressionlevels significantly increased from 12 hours to 72 hours. A peak was found at 24 hours after IP. However, there were no significant changes in both membrane translocation of cPKC?, ?Ⅰ、?Ⅱand protein expression of cPKC?Ⅰ, ?Ⅱ in retina of rats following IP.Conclusion The enhanced cPKC? membrane translocation, the increased cPKC? and ? protein expressions might be involved in the onset and sustain of retinal IPC in rats respectively.

Objective To develop a reliable and reproducible model of ischemic stroke .Methods Male C57BL/6J mice were randomly divided into three groups:three-vessel occlusion (3VO, 15-min temporary occlusion of the bi-lateral common carotid arteries superimposed on a permanent occlusion of the right distal middle cerebral artery , n=20 ) , suture MCAO control ( n=20 ) , and sham-operated group ( n=10 ) .At 24 h functional outcome was as-sessed using corner test , cerebral ischemic lesion was determined by TTC staining ( n=10 ) , and then the surviral rate was measured till day 7 (n=10).Results The 3VO group got significantly lower neurological deficit score as compared to sham-operated group ( -0.7 vs -0.04 , P<0.01 ) .Infarct volume of mice in 3 VO group was 17.6%±1.6%, while 42.6%±15.0%in MCAO group ( n=10 ) .The 3 VO group got a significantly increased 7-day survival rate compared to suture MCAO group (90%vs 60%, n=10).Conclusions The present three-ves-sel occlusion model is stable and reliable for research on ischemic stroke .

Objective To investigate the effects of VCAM-1 on migration and invasion of glioma cell lines . Methods The techniques of lentivirus pSGU6/GFP/Neo-based VCAM-1 shRNA and EF1 a-GFP/puro-based VCAM-1 expression vector, the scratch wound healing migration and transwell invasion assays , and the Western blot and cell staining were applied to observe the effects of VCAM-1 expression levels on migration and invasion of glioma cell line cells.There are four groups in T98G cells including control, vector, scramble and shRNA-VCAM-1 groups and three groups in U251 cells covering control, vector and VCAM-1 overexpressed groups ( n=6 per group) .Results The stabled glioma cell lines of T98 G cells with down-regulated VCAM-1 and U251 cells with VCAM-1 overexpression were established by using lentivirus-based VCAM-1 shRNA and expression vector.The ability of scratch wound healing (migration activity) decreased significantly (P<0.01) in T98G cells with lower VCAM-1 expression levels, while the migration activity was obviously improved in U251 cells with overexpressed VCAM-1 ( P <0.05 ) .Similarly, the invasion ability was significantly inhibited ( P <0.05) in T98G cells with silenced VCAM-1, as well as VCAM-1 overexpression could enhance the invasion ability of U251 cells ( P<0.01 ) .Conclusions VCAM-1 improves the migration activity and invasion ability of human glioma cell line cells.