Sirt1-ROS-TRAF6 Signaling-Induced Pyroptosis Contributes to Early Injury in Ischemic Mice / 神经科学通报·英文版

Stroke is an acute cerebro-vascular disease with high incidence and poor prognosis, most commonly ischemic in nature. In recent years, increasing attention has been paid to inflammatory reactions as symptoms of a stroke. However, the role of inflammation in stroke and its underlying mechanisms require exploration. In this study, we evaluated the inflammatory reactions induced by acute ischemia and found that pyroptosis occurred after acute ischemia both in vivo and in vitro, as determined by interleukin-1β, apoptosis-associated speck-like protein, and caspase-1. The early inflammation resulted in irreversible ischemic injury, indicating that it deserves thorough investigation. Meanwhile, acute ischemia decreased the Sirtuin 1 (Sirt1) protein levels, and increased the TRAF6 (TNF receptor associated factor 6) protein and reactive oxygen species (ROS) levels. In further exploration, both Sirt1 suppression and TRAF6 activation were found to contribute to this pyroptosis. Reduced Sirt1 levels were responsible for the production of ROS and increased TRAF6 protein levels after ischemic exposure. Moreover, N-acetyl-L-cysteine, an ROS scavenger, suppressed the TRAF6 accumulation induced by oxygen-glucose deprivation via suppression of ROS bursts. These phenomena indicate that Sirt1 is upstream of ROS, and ROS bursts result in increased TRAF6 levels. Further, the activation of Sirt1 during the period of ischemia reduced ischemia-induced injury after 72 h of reperfusion in mice with middle cerebral artery occlusion. In sum, these results indicate that pyroptosis-dependent machinery contributes to the neural injury during acute ischemia via the Sirt1-ROS-TRAF6 signaling pathway. We propose that inflammatory reactions occur soon after oxidative stress and are detrimental to neuronal survival; this provides a promising therapeutic target against ischemic injuries such as a stroke.

VCAM-1 improves migration and invasion of human glioma cell lines / 基础医学与临床

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.

A modified three-vessel occlusion mouse model with ischemic stroke / 基础医学与临床

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 .

Epidural ropivacaine block increasing anesthetic potency of propofol / 基础医学与临床

Objective To investigate the effects of epidural ropivacaine block combined with propofol intravenous anesthesia on CaMKⅡ and ERK1/2 total protein (T-CaMKⅡ and T-ERK1/2) and phosphorylation(p-CaMKⅡ and p-ERK1/2) levels in the hippocampus and cortex of rats.Methods Rats were randomly assigned to three groups: group P(control,propofol intravenous anesthesia),group PS(propofol and epidural normal saline) and group PR(propofol and epidural 0.5% ropivacaine).Anesthesia were performed in 72 h after epidural catheter placement.The rats in group PR received 70 μL of 0.5% ropivacaine to achieve epidural block.1% propofol was infused through rats caudal vein.Propofol dosage for anesthesia induction was 12 mg/kg,for anesthesia maintenance was 40 mg/(kg·h).Before the rats were decapitated,the depth of anaesthesia was assessed as either light anesthesia or deep anesthesia by checking of pinch withdrawalreflex,eyelid reflex and spontaneous rapid whisking of the vibrissae after propofol continuous infusion for 1 h.T-CaMKⅡ/T-ERK1/2 and p-CaMKⅡ/p-ERK1/2 in hippocampus and frontal cortex were examined by Western blot.Results 7 rats were assessed as light anesthesia and one rat as deep anesthesia in group P;6 rats were assessed as light anesthesia and 2 rats as deep anesthesia in group PS;in group PR,1 rat was assessed as light anesthesia and 7 rats as deep anesthesia.Significant differences were seen among three groups (P<0.05).In hippocampus of rats,p-CaMKⅡ(Thr286)43.7%±8.8% and p-ERK1/2 32.4%±7.9% in group PR were significantly lower than those in group P (100%,P<0.05).Conclusions Epidural ropivacaine block may strengthen the depth of anesthesia achieved with propofol intravenous anesthesia.The decrease of p-CaMKⅡ(Thr286) and p-ERK1/2 in hippocampus of rats may explain the effects of epidural block.

Dynamic expression of conventional protein kinase C-γ with development of visual cortex and effect of monocular deprivation on it in mice / 中华实验眼科杂志

Background Plasticity of visual system is one of the mechanisms of deprivation amblyopia.Our previous study showed that synapsin plays a role during visual developmental plasticity,and conventional protein kinase C-γ (cPKC-γ) probably is one of upstream kinases of synapsin.However,whether or how the cPKC-γ plays its effects on visual developmental plasticity is below understood.Objective This study was to investigate the dynamic expression of cPKC-γ in visual cortex of normal mice and explore the effects of abnormal visual experience on cPKC-γ expression.Methods The bilateral visual cortex tissues were obtained from 36 clean C57BL/6 mice at postnatal (P) 7,14,21,28,35,42 days respectively and 6 mice for each for the researching of cPKC-γ dynamical expression in visual cortex over aging.Other 24 C57BL/6 mice were randomized into developmental phase group and adult phase group,12 for each group.The monocular deprived (MD) models were established by suturing the upper and inferior eyelides in P14 mice for 14 days in 6 mice in the developmental phase group and 6 healthy mice served as controls,and MD models were established in the same way in 6 P60 mice in the adult phase group,and the same aged mice (6 mice) were used as controls.The mice were sacrificed and bilateral visual cortexes were obtained.The expression of cPKC-γ protein in the visual cortex was quantitatively detected using Western blot assay.The study protocol was approved by Ethic Committee of Tongren Eye Hospital.The use and care of the experimental mice followed the ARVO Statement.Results The cPKC-γ protein was faintly expressed in visual cortex in normal P7 mice,with the related expressing level of (39.74± 11.22)％ and (40.78± 10.37)％ in the left and right cortex,respectively.The expressing level of cPKC-γ protein was gradually increased over aging,with the peak value of (138.68±15.73)％ and (138.47±23.48)％ in P21 mice.A significant difference was found in the expression of cPKC-γ protein in various ages of mice (Fage =57.174,P =0.000),and the expression of cPKC-γ protein was not significantly different between the left and right visual cortexses (Flateral =0.059,P =0.809).No significant differences were found in the expression of cPKC-γ protein on bilateral visual cortexes among the mice of the developmental phase group and adult phase group (Fage =1.798,P =0.159) or among the MD group and normal control group (Fgroup =0.104,P=0.749).Conclusions The dynamic expression of cPKC-γ in the visual cortex of normal mice presents a consistant tend with the aging and development of visual critical period.MD does not affect the expression of cPKC-γ protein in bilateral visual cortexes.Further researches should be performed in the activity of cPKC-γ protein in MD mice.

Impaired sustained attention function of heroin abusers in attention switching / 中国神经精神疾病杂志

Objective To investigate the effect of heroin abuse on attention switching. Methods Thirty-six Heroin abusers (33 males, 3 females) and 36 controls (32 males, 4 females) were enrolled in the study. Their cognitive function was tested by using the Switching Task, including Sustained Attention trials and Switching Attention trials. The reaction time and accuracy were recorded separately by the computer. Results The accuracy or reaction times were not signifi-cantly different between Switching Attention trial and Sustained Attention trial in heroin abusers, suggesting a lower Switch Costs value compared to the healthy controls [(19.7 ± 66.8) ms vs. (85.1 ± 92.4) ms]. The healthy controls showed faster reaction speed [Sustained Attention trial (695.3 ± 95.9) ms vs. Switching Attention trial (780.3 ± 93.3) ms, P<0.05] and higher accuracy [Sustained Attention trial (98.0%±2.2%) vs. Switching Attention trial (93.8%±5.0%), P<0.05] under the Sustained Attention trial. Compared with the healthy controls, the heroin abusers showed slower reaction speed [(791.6 ± 74.3) ms vs. (695.3±95.9) ms, P<0.05] and lower accuracy [(92.5%±8.4%) vs. (98.0%±2.2%), P<0.05] in Sus-tained Attention trial, but not in Switching Attention trial. Conclusions The present study has revealed absence of Switch Costs in heroin abusers, which may be related to the damage of heroin abusers in their Sustained Attention function.

The effect of anxiety on attentional orienting of heroin addicts / 中国神经精神疾病杂志

Objective To investigate the effect of state anxiety and trait anxiety on attentional orienting of heroin addicts. Methods State anxiety and trait anxiety was measured by State-Trait Anxiety Inventory (STAI). Forty heroin ad?dicts (36 males and 4 females) and 40 healthy controls (36 males and 4 females) participated in cue-target task. Atten?tional orienting and reorienting were measured in valid cue trials and invalid cue trails. Results Heroin addicts had sig?nificantly greater state anxiety [(42.65 ± 6.58) vs. (36.60 ± 8.91)] and trait anxiety [(44.43 ± 7.67) vs. (37.00 ± 8.63)] values than controls (P<0.05). The state anxiety was significantly correlated with orientation RT difference (r=-0.259, P=0.020) and disengaging/reorientation RT difference (r=0.333, P=0.003) in heroin addicts. Trait anxiety was also significantly cor?related with orientation RT difference (r=-0.248, P=0.026) and disengaging/reorientation RT difference (r=0.356, P=0.001) in heroin addicts. Conclusion Heroin addicts have significantly greater anxiety than healthy controls. Both their state anxiety and trait anxiety are associated with attentional orienting and disengaging/reorienting.

Role of CaMK Ⅱ in primary somatosensory area and hippocampi in reduction of remifentanil-induced hyperalgesia by lidocaine in rats / 中华麻醉学杂志

Objective To investigate the role of calcium/calmodulin-dependent protein kinase Ⅱ (CaMK Ⅱ) in the primary somatosensory area (S1 area) and hippocampi in reduction of remifentanil-induced hyperalgesia by lidocaine in rats.Methods One hundred fifty-six male Sprague-Dawley rats,aged 8-10 weeks,weighing 240-260 g,were randomly allocated into 4 groups using a radom number table:control group (group C,n=6),remifentanil group (group R,n=50),lidocaine group (group L,n=50),and remifentanil+lidocaine group (group RL,n =50).Remifentanil was given as a bolus of 6 mg/kg followed by an 2 h infusion of 2.4 μg · kg-1 · min-1 in group R.Lidocaine was given as a bolus of 6 mg/kg followed by an infusion of 200 μg · kg-1 · min-1 for 2 h in group L.In group RL,drug administration was similar to those previously described in R and L groups.The mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were measured before administration and at 0.5,2,5 and 24 h after the end of administration.The rats were then sacrificed immediately after administration and at 0.5,2,5 and 24 h after the end of administration in R,L and RL groups,or at the corresponding time point in group C.The S1 area and hippocampi were isolated for determination of phosphorylated CaMK Ⅱ (p-CaMK Ⅱ) expression by Western blot.Results Compared with the value before administration,the MWT was significantly decreased at 0.5 and 2 h after the end of administration (P＜0.05),and no significant change was found in TWL at each time point after the end of administration in R,L and RL groups (P＞0.05).Compared with group C,p-CaMK Ⅱ expression in the S1 area and hippocampi was significantly up-regulated immediately after administration and at 0.5 and 2 h after the end of administration in group R (P＜0.05).Compared with group R,p-CaMK Ⅱ expression in the S1 area and hippocampi was significantly down-regulated immediately after administration and at 0.5 and 2 h after the end of administration in group RL,and p-CaMK Ⅱ expression in the S1 area was significantly down-regulated immediately after administration,and at 0.5 and 2 h after the end of administration,and p-CaMK Ⅱ expression in the hippocampi was down-regulated immediately after administration,and at 0.5,2 and 24 h after the end of administration in group L,and MWT was increased at 0.5 and 2 h after the end of administration in groups L and RL (P＜0.05).There was no significant difference in TWL at each time point among the three groups (P＞0.05).Conclusion The mechanism by which lidocaine mitigates remifentanil-induced hyperalgesia is associated with inhibited activity of CaMKII in the S1 area and hippocampi of rats.

Study on numerical distance effect in amblyopic children aged from 6 to 13 / 中华行为医学与脑科学杂志

Objective To investigate numerical distance effect of numerical processing in amblyopic children aged from 6 to 13. Methods 46 amblyopic children and 43 control children with normal sight were divided into 3 groups respectively:7Y group (6~8 years old),9Y group (9~10 years old) and 12Y group (11~13 years old). The numerical comparison task (judging the magnitude of number) was used in this study and error rate( ER) and reaction times ( RTs) were recorded. Results ( 1) RTs of judging the num-ber 4 (7Y group (995±100)ms,9Y group (964±141)ms,12Y group (701±125)ms) were significant lon-ger(P5).The develop-ment of numerical processing in amblyopic children is slower than that in children with normal sight.

Changes of folate receptor -α protein expression in human gliomas and its clinical relevance / 中华外科杂志

<p><b>OBJECTIVE</b>To study the expression level of folate receptor α (FR-α) in glioma tissue and its clinical significance.</p><p><b>METHODS</b>Forty-eight human glioma specimens were collected from patients who underwent surgery from March 2012 to March 2013. These specimens were as follows:12 cases of glioblastoma (WHO IV), 6 cases of astrocytoma of each malignancy grade(WHO II, III), 6 cases of oligodendroastrocytoma of each malignancy grade (WHO II, III), 6 cases of oligodendroglioma of each malignancy grade (WHO II, III ). In addition, 6 cases of normal brain tissue resected from brain traumatic patients were taken as negative control, and one case of placental tissue (had got the consent of the parents and their families) was taken as positive control. The expression level of FR-α in tumor tissues was evaluated by Western blot analysis. The results of Western blot analysis were analyzed by t-test. The expression level of FR-α and Ki-67 in tumor tissues was evaluated immunohistochemistry, the results were analyzed by Kruskal-Wallis test and Nemenyi test. The correlation between the expression level of FR-α and cell proliferation index Ki-67 was analyzed by Pearson correlation analysis.</p><p><b>RESULTS</b>Western blot analysis showed that the FR-α was not expressed in normal brain tissue and oligodendroglioma tissue, but highly expressed in astrocytoma, oligodendroastrocytoma and gliomablastoma. The expression level in WHO III astrocytoma was significantly higher than in WHO II (t = 4.497, P < 0.05). FR-α was also highly expressed in oligodendroastrocytoma and its expression level in WHO III was also significantly higher than in the WHO II (t = 2.876, P < 0.05). Foremore, immunohistochemistry analysis also showed that FR-α was not expressed in oligodendroglioma, but expressed in astrocytoma, oligodendroastrocytoma and gliomablastoma. The positive rate of FR-α of WHO III was significantly higher than the WHO II astrocytoma(57.8% ± 2.2% vs. 45.7% ± 2.3%,χ(2) = 3.871, P = 0.034). In oligodendroastrocytoma, the positive rate of FR-α of WHO III was significantly higher than the WHO II(56.5% ± 5.4% vs. 37.1% ± 5.2%,χ(2) = 4.454, P = 0.021). Moreover, the expression level of FR-α in gliomablastoma was highest in all histological types of gliomas, the positive rate of FR-α was up to 65.0% ± 4.5%. Pearson correlation analysis showed that the positive rate of FR-α was positively correlated with Ki-67 index (r = 0.903, P < 0.05).</p><p><b>CONCLUSIONS</b>FR-α is expressed in astrocytoma, oligodendroastrocytoma and glioblastoma, and the expression level of FR-α is positively correlated with malignancy grade and Ki-67 index. Therefore, FR-α may be applied as a special target for diagnosis and treatment of glioma.</p>

Expression of synapsin in visual cortex of developing mice / 中华实验眼科杂志

Background The treatment timing and method of amblyopia rely on the plasticity of visual system.Synapsin is a family of presynaptic terminal specific protein.Its role in visual developmental plasticity is below understood.Objective To investigate the dynamic expressions of synapsin (T-synapsin),and phosphorylation of synapsin (p-synapsin Ⅰ a/b) in visual cortex of normal mice and further explore the role of synapsin in plasticity of visual system.Methods Forty-two clean neonatal C57BL/6 mice were collected.The mice were sacrificed at postnatal 7,14,21,28,35,42,60 days respectively to obtain the tissue samples of visual cortex.Expression levels of T-synapsin and p-synapsin in the visual cortex following the ageing were quantitatively detected using Western blot assay.Results The expression of synapsin in normal mice showed a dynamic increase with the ageing.The T-synapsin Ⅰ a/b/β-actin value in visual cortex was (21.32 ± 3.27) ％,(56.27 ± 10.18) ％,(77.05 ± 10.05) ％,(83.75±10.52) ％,(94.69±11.46)％,(98.75±5.86) ％ of adults mice (postnatal 60 days,P60) in the mice of postnatal 7,14,21,28,35,42 days,respectively,showing a significant difference among them (F =69.538,P ＜ 0.001).Compared with the adult mice,the T-synapsin Ⅰ a/b/β-actin value in the mice of P7,P14,P21,P28 was significantly lower (all at P＜0.05),but no significant difference was found between P35 and P60,P42 and P60 (P =0.280,0.798).The development trend of different synapsin subtypes,such as T-synapsin Ⅰ a/b,T-synapsin Ⅱ a,T-synapsin Ⅱ b and T-synapsin Ⅲ a,was not quite the same during the ageing.The expression of T-synapsin Ⅱ a and Ⅲ a increasing more slowly with development,and kept increasing until P60.Significant differences were found among various age of mice in T-synapsin Ⅱ a,Ⅱ b,Ⅲa respectively(F =42.492 55.595,39.172,all at P＜0.001).The p-synapsin Ⅰ a/b level in the visual cortex elevated with the ageing of the mice,and that peaked in P21 mice,which was (2.86±0.17) times more than that in adult mice.After that,the expression level of p-synapsin Ⅰ a/b dropped rapidly.A significant difference was found in the p-synapsin Ⅰ a/b expression among different ages of mice (F =22.620,P ＜ 0.001).Conclusions Synapsin level in visual cortex presents a developmental change which correlated with the onset and decline of the critical period.Synapsin is probably involved in the regulation of neural plasticity in visual cortex in critical period.

The effects of distraction on tourniquet pain in healthy subjects / 重庆医学

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 .

The different durations of methamphetamine abuse influenced abusers' spatial processing function / 中华行为医学与脑科学杂志

Objective To investigate the effects of different durations of methamphetamine abuse on spatial cognitive processing.Methods The mental rotation task was used to evaluate the spatial processing function of 35 methamphetamine abusers and 30 healthy subjects.The methamphetamine abusers are divided into 2 groups according to their abuse durations,group 1 contains 16 abusers with an average duration of 1 year and group 2 contains 19 abusers with an average duration of 3 years.Participants were asked to judge the right hand/food or the left hand/food of the experimental stimuli at difference angles.The reaction time (RT) and the accurate rate (AR) of the mental rotation task were recorded.Results Compared with the control group,RT of group 1 methamphetamine abusers was longer at the angle of 0°((1469±318)ms) and 180°((1718±412)ms) (P＜0.05),RT of group 2 was longer at the angle of 0°((1466±243) ms),60°((1497±294) ms) and 180°((1708±288) ms) (P ＜0.05).And the AR of methamphetamine abusers was higher at every angle (P＜0.05).Conclusion Long-term dependence on methamphetmine can damage the abuser's spatial processing function.

Effect of delayed preconditioning with morphine on ischemic cerebral injury in mice and the role of classical protein kinase C / 中华麻醉学杂志

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.

The effects of tourniquet pain on attention bias towards pain-related words and its gender difference / 中华行为医学与脑科学杂志

ObjectiveTo investigate the effect and gender differences of experimental moderate pain on attentional bias towards pain-related words.MethodsWith the tourniquet around the left upper limb,32 healthy college students were asked to perform two dot-probe tasks including five kinds of pain-related words,one task with the tourniquet deflating and the other task with the tourniquet inflating to 200 mm Hg pressure.The reaction time (RT) and the error rate of recognition task were recorded.The pain intensity and distress were recorded too.Results ( 1 ) Compared with no pain condition ( RT of affective and social-threat words were (507 ± 78 ) ms,(504 ±89 )ms respectively),the react time was shorter in female in the condition of tourniquet pain,particularly for affective and social-threat words (the RT were (464 ± 79 )ms,(465 ± 72 )ms respectively,F ( 1,4) =19.157,P ＜0.01 ),but not in male.( 2 ) In condition of no pain,the reaction time of male ( (456 ± 59) ms ) was shorter than that of female (504 ± 79ms).However,in the condition of pain,the significant main effect was not found.ConclusionThe reacting time of female was shorter in condition of moderate tourniquet pain,which imply that female showed significant attentional bias towards affective and social-threat words.

The effect of experimental pain on attentional bias in a pictorial dot-probe task / 中华物理医学与康复杂志

Objective To investigate the effect of moderate pain on attentional bias towards emotional pictures among healthy subjects.Methods Thirty-two healthy college students aged from 17 to 26 (21.8±2.2;16 males and 16 females) participated in this study.A tourniquet was tied to each subject's left upper arm 1 to 2cm above the cubits horizontal grain.Pain was inflicted by inflating the tourniquet,and the pressure was maintained at 26.66kPa.While tourniquet was inflated (with pain) or not (no pain),each subject was asked to finish a pictorial dot-probe task with three kinds of pictures-emotionally positive,negative and neutral.In experiment 1,subjects performed the dot-probe tasks with the contralateral hand while the tourniquet was tied on the left upper arm without inflation.In experiment 2 the tourniquet was inflated until the subject completed the dot-probe task (for about 10min).The reaction times (RTs) and the error rates (Ers) in the recognition task were recorded,and the intensity of the subject's pain and discomfort were measured using a verbal rating scale.Results The subjects reported moderate to severe pain with the tourniquet inflated.The RT and ER data were analyzed using two-way analysis of variance (ANOVA) which showed a significant difference between the average RTs of the males (482±73ms without pain and 466±82ms with pain) and those of the females (536±90ms without pain and 519±100 ms with pain).The average ER was significantly different between the pain (2.38±1.49)％ and no pain (1.09±0.82)％ conditions in both groups.Holn-Sidak multiple comparison testing showed significant differences in both groups' average ER between the negative picture (3.81±1.73)％ and the positive picture (1.66±0.97)％,and between the negative and neutral pictures (1.68±0.8) ％ in the pain condition.Mild attentional avoidance was observed with the positive [pain condition (-5.1±4.8) ms and no pain (-4.6±4)ms] and negative pictures [pain condition (-3.43±6) ms and no pain (-0.79±4.1)ms],but no significant difference was found between the pain and no pain conditions.Conclusion The error rate in a pictorial dot-probe task is influenced by pain,especially with negative pictures.

Measurement of skin color in 600 individuals from three geographical regions in China / 中华皮肤科杂志

Objective To assess the difference in human skin color among different geographical regions including Northern,Eastern and Sourthern China.Methods Six hundred individuals from Northern (Beijing),Southern (Shenzhen and Dongguan in Guangdong province) and Eastern (Changzhou in Jiangsu province and Yantai in Shandong province) China were included in this study.A Multi Probe Adapter MPA 9 device was used to measure the skin color of 4 body sites,including forehead,left cheek,left medial forearm,and the back of left hand,of these subjects.The Commission International de l'Eclariage (CIE) L*a*b* values were recorded and individual typological angle (ITA) was calculated.Skin color was rated on a scale from 1 to 6 which was proposed by Chardon et al.The SPSS 16.0 software was used to process these data,and the skin color grade distribution was compared by chi-square test among different geographical regions and body sites.Results Among the 2400 tested sites from 600 subjects,2.96％ were rated as grade Ⅰ,39.88％ as grade Ⅱ,36.58％ as grade Ⅲ,16.96％ as grade Ⅳ,3.63％ as grade V,none as grade Ⅵ.In Northern China,Ⅰ and Ⅱ were the most prevalent grade of skin color (530 sites),while ⅣV and Vwere relatively rare (37 sites); in Eastern China,grade Ⅲ predominated (335 sites),which was followed by ⅣV and V (110 sites); in Southern China,the most common grade was Ⅳ (267 sites),while the rarest grade was Ⅰ (2 sites).The back of the left hand seemed to be the blackest,while the left cheek and left medial forearm the whitest,with significant differences in the distribution of skin color grade between these 4 tested body sites (x2 =106.00,P ＜ 0.01).Conclusions Skin color varies among different geographical regions in China,and among different sites on the human body.

Factors affecting the protective effect of morphine preconditioning on murine hippocampal neurons against anoxia-reoxygenation injury / 中华麻醉学杂志

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.

Increased phosphorylation of CaMK Ⅱ in cerebral tissues from hypoxially preconditioned mice / 基础医学与临床

Objective To explore the role of calcium/calmodulin-dependent protein kinase Ⅱ ( CaMK Ⅱ ) in the development of cerebral hypoxic preconditioning(HPC). Methods Healthy male BALB/c mice were randomly divided into 7 groups as follows; normoxic control (H0) , early(H1～H4) and delayed (H5～H6) hypoxically preconditioned mice groups. SDS-PAGE, Western blot and Gel Doc imagine systems were applied to quantitatively analyze the level of CaMK Ⅱ phosphorylation and protein expression level in the brain of mice. Results Compared with H0 group, the phosphorylation level of CaMK Ⅱ increased in cortex and hippocampus of mice in H3～H5 hypoxically preconditioned groups (P<0.05). However, there was no significant changes in total CaMK Ⅱ protein expression in cortex and hippocampus of hypoxic preconditioned mice. Similarly, enhanced p-Thr286 CaMK Ⅱ was also observed in the hippocampus and cortex of mice by immunostaining following hypoxic exposures (H3 and H6). Conclusion The increased phosphorylation of CaMKⅡ may be involved in the development of cerebral HPC in mice.

Identification of nPKCε-interacted proteins in the cortex of hypoxic preconditioned mice / 基础医学与临床

Objective Identify novel protein kinase Cε(nPKCε)-interacted proteins in the cortex of hypoxic preconditioned mice.Methods Immunoprecipitation (IP) and two-dimensional electrophoresis (2-DE) combining with ImageMaster 2D Platinum software were applied to analyze the differential expressions of nPKCe-interacted proteins;the target protein spots were identified by matrix-associated laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS) and Western blot.Results Compared with control group,there were 34 upregulated protein spots and 20 downregulated protein spots in cytosolic fraction,while 27 upregulated prtein spots and 28 downregulated protein spots were determined in particulate fraction of cerebral cortex of HPC mice.The levels of nPKCε-interacted HSP 70 and 14-3-3γ/protein expressions increased significantly in both cytosolic and particulate fractions;but the protein level of nPKCε-interacted HSP60 increased only in particulate fraction of cerebral cortex of HPC mice.Conclusion nPKCε might be involved in the development of cerebral HPC via the regulation of its interacted proteins such as HSP60,HSP70 and 14-3-3γ.