of apoptosis-related gene.

Objective To investigate the role of phosphatidylinositol 3?kinase∕protein kinase B∕glycogen synthase kinase?3β ( PI3K∕Akt∕GSK?3β) signaling pathway in hydrogen?induced inhibition of neuronal ap?optosis induced by focal cerebral ischemia?reperfusion ( I∕R) in rats. Methods One hundred and twenty
healthy adult male Sprague?Dawley rats, weighing 200-220 g, were divided into 5 groups ( n=24 each) u?sing a random number table: sham operation group ( group S); cerebral I∕R group ( group I∕R); hydrogen group (group H2); LY294002 (specific PI3K inhibitor) group (group LY); LY294002+hydrogen group ( group LY+H2 ) . Focal cerebral I∕R was induced by occlusion of the middle cerebral artery for 1 h followed by 24 h reperfusion. In H2 and H2+LY groups, the animals inhaled 67% H2+33% O2 for 2 h starting from onset of reperfusion, and then inhaled H2 for 2 h every 6 h. In LY and LY+ H2 groups, LY294002 ( 10 mmol∕L) 10 μl was injected into the lateral cerebral ventricle at 10 min before reperfusion. Neurologic defi?cit was evaluated and scored ( NDS) at 24 h of reperfusion. The rats were then sacrificed, and the brains were removed for measurement of the cerebral infarct size ( by TTC staining) and apoptosis in cortical neu?rons ( by TUNEL) and for determination of the expression of Akt in the ischemic cerebral cortex, phospho?rylated Akt ( p?Akt) , GSK?3β and phosphorylated GSK?3β ( p?GSK?3β) and Bcl?2 and Bax positive cell count in the ischemic cerebral cortex ( by immuno?histochemistry) . The apoptosis index ( AI) , p?Akt∕Akt ratio and p?GSK?3β∕GSK3?β ratio were calculated. Results Compared with group S, the NDS, cerebral infarct size, AI, p?Akt∕Akt ratio, p?GSK?3β∕GSK?3β ratio and Bax positive cell count were significantly increased, and the Bcl?2 positive cell count was significantly decreased in group I∕R ( P<0?05) . Compared with group I∕R, the NDS, cerebral infarct size, AI and Bax positive cell count were significantly de?creased, and the p?Akt∕Akt ratio, p?GSK?3β∕GSK?3β ratio and Bcl?2 positive cell count were significantly increased in group H2 ( P <0?05) , and no significant changes were found in the parameters mentioned a?bove in LY and LY+H2 groups ( P>0?05) . Compared with group H2 , the NDS, cerebral infarct size, AI and Bax positive cell count were significantly increased, and the p?Akt∕Akt ratio, p?GSK?3β∕GSK?3βratio and Bcl?2 positive cell count were significantly decreased in group LY+H2 ( P<0?05) . Conclusion The mechanism by which hydrogen inhibits focal cerebral I∕R?induced neuronal apoptosis is associated with the activation of PI3K∕Akt∕GSK?3β signaling pathway in rats.

Objective To investigate the effects of brain-derived neurotrophic factor (BDNF) pretreatment on neuron apoptosis and the expression of Bcl-2 and Bax protein following global cerebral ischemia-reperfusion (I/R) injury in gerbils. Method Forty-eight mongolian gerbils were randomly divided into six groups in equal number (n = 8): normal control group (group C), ischemia-reperfusion group (group I/R) and four BDNF pretreatment groups according to various lengths of time from BDNF pretreatment to ischemia-reperfusion. The BDNF pretreat-ment was carried out in gerbils with lateral ventricular injection of BDNF 0.5μg 6 h, 12 h,24 h and 48 hours be-fore cerebral ischemia, and those gerbils assigned into PR6, PR12, PR24 and PR48 groups. The global cerebral is-chemia-reperfusion was induced by occlusion of bilateral common carotid arteries for 20 minutes and then the arter-ies were released for 24 hours reperfusion. The confirmation of global cerebra ischemia was evidenced by the ap-pearance of mydriasis and disappearance of light reflex and righting reflex. Twenty-four hours later, all gerbils including those of control group were sacrificed and a piece of tissue was taken from frontal cortex just behind the optic chiasma 1～4 millimeter for making paraffin sections. Neuron apoptosis was identified by using TUNEL and immunohischemistry was used to detect the expression of Bcl-2 and Bax protein in cerebral cortex. The data were analyzed by using analysis of variance. Results There were no apoptotic cells, and expression of Bcl-2 and Bax protein positive cells found in group C. Neuron apoptosis in brain cortex was detected in I/R group and BDNF pre-treatment groups. The indexes of neuron apoptosis in BDNF pretreatment groups were markedly lower than those in group I/R (P < 0.01). Compared with group I/R, the index of expression of Bcl-2 protein positive cells was in-creased significantly in BDNF pretreatment groups (P = 0.005), while the index of expression of Bax protein posi-tive cells were decreased significantly (P < 0.01 in all groups). Among 4 BDNF pretreatment group, the lowest apoptosis index and lowest of expression of Bax protein positive cells were found in PR6 and PR12 BDNF pretreat-ment groups (P = 0.0056 and 0.001, respectively). Conclusions Different time windows of BDNF pretreatment can decrease the neuron apoptosis in different degree, and protect brain against cerebral ischemia-reperfusion injury significantly. Among BDNF pretreatment time windows, pretreatment of 6 hours and 12 hours are the better ones.The mechanism of protection of BDNF pretreatment may be attributed to inducing Bcl-2 protein expressions and in-hibiting Bax protein expressions, and thereby inhibiting neuron apoptosis.

ObjectiveTo investigate the effects of postconditioning with Shenmai-injectio on myocardial ischemia-reperfusion (I/R) injury in rats.MethodsThirty-six healthy male SD rats aged 10-12 weeks weighing 240-260 g were randomly divided into 3 groups ( n =12 each):sham operation group (group S) ; myocardial I/R group and Shenmai-injectio postconditioning group (group SPO).Myocardial I/R was produced by ligation of the left anterior descending branch of coronary artery for 30 min followed by 120 min reperfusion in groups I/R and SPO.In group SPO Shenmai-injectio 9 ml/kg was injected iv at the end of 30 min ischemia.Blood samples were collected from abdominal aorta at the end of 120 min reperfusion for determination of serum CK activity and cTnI concentration.The animals were then sacrificed.Myocardial specimens were obtained for microscopic examination,detection of apoptosis and determination of myocardial Bcl-2 and Bax protein expression ( by immuno-histochemis-try).ResultsMyocardial I/R significantly increased serum CK activity,cTnI concentration,apoptotic index (percentage of apoptotic cells) and Bax protein expression and decreased Bcl-2 protein expression in group I/R as compared with group S.Shenmai-injectio postconditioning significantly attenuated I/R-induced above changes and ameliorated histo-pathological damage in group SPO as compared with group I/R.ConclusionShenmai-injectiopostconditioning can reduce myocardial I/R injury by up-regulating Bcl-2 expression and down-regulating Bax expression,leading eventually to reduction in apoptosis.

Objective Boy sexual development is affected not only by age, but also by environment factors .The present study was to survey and evaluate the status of sexual development in boys from Zhengzhou area and to explore their influencing factors. Methods The height, weight, waist circumference, hip circumference, penis and testicles in 3779 boys aged from 6 to 13 years old in Zhengzhou were measured .The penis length and testicular volume of different ages and their influencing factors were compared, and the beginning age and sexual development characteristics were explored .The development of genitalia and pubes were evaluated by Tanner staging criteria and standards, which were divided into five stages. Results There were 3779 boys, including 395 overweight cases (10.45%) and 208 obesity cases (5.50%).The testicular volume and penis length increased by age, which increased slowly at the age of 6 to 11, followed by a rapid increase after the age of 11 year old.Testicular volume increased more rapidly than that of penis length. The medians quartile of testicular volume in overweight group ,obese group and normal group were 2.50 mL, 2.50 mL, 2.00 mL, 2.00 mL,2.00 mL,1.00 mL respectively .The testicular volume in overweight group and obese group were significantly greater than that in nor -mal group (P<0.01) and there were no difference between overweight group and obese group .The penis length in overweight group and obese group were significantly less than that in normal group (P<0.01), and there was no difference between overweight group and obese group.Testicular volume and penis length were positively correlated with age, height, weight, body mass index, waist circumference and hip circumference; the correlation between testicular volume and height, penis length and age were the strongest. The age of development of testicle (average 12.02 years) in urban area was later by 2.73 years than that in the rural area (average 9.9 years), the difference was statistically significant (P<0.001).The age of pubic development (P2) and maturity (P5) was (12.34 ±0.83) years and (12.39 ±1.24 ) years.The incidence of precocity in boys was 0.40%. Conclusion The sexual development of boys from Zheng-zhou area is similar to the results of large scale survey from China .Attention should be paid to the effect of obesity on sexual develop-ment, and early intervention should be given, the occurrence of adverse events of sexual development can be avoided .

Objective To investigate the correlation of anthropometric indicators and blood pressure in children. Methods A total of 6 790 children aged 6 to 13 years were inspected by random sampling. The body mass index (BMI), waist circumference (WC), hip circumference (HC), and blood pressure were measured. The waist/hip ratio (WHR) and waist/height ratio (WHtR) were calculated. The data wrer analyzed by SPSS16.0 statistical software. Results The detection rate of hypertension in children was 5.57%. After controlling for age, both in male and female children, the BMI, WC, HC, WHR, WHtR and systolic blood pressure showed a significant positive correlation by partial correlation analysis (all P<0.05). Both in male and female children, the BMI, WC, HC, WHR, and WHtR were significantly higher in children with hypertension than those in children with normal blood pressure (all P<0.05). Among all subjects, 280 children (4.12%) were obese, 622 children (9.16%) were overweight. The detection rates of hy-pertension were significantly different among obese, overweight and normal weight children (P<0.01). The detection rate of hypertension was significantly higher in obese children than that in overweight and normal weight children. The systolic and diastolic blood pressure were all significantly higher in obese and overweight children than that in normal weight children (P<0.05). Conclusion The hypertension prevalence of children aged 6 to 13 years in Zhengzhou is in the low to median level in the same age groups. The BMI, WC, HC, WHR, WHtR are signiifcantly correlated with blood pressure in both gender, and the correlation is the most obvious in HC.

Objective To investigate the effects of inhaling high concentration of hydrogen gas on the expressions of endoplasmic reticulum stress(ERS) related protein glucose regulated protein 78 (GRP78),Caspase-12 and the neural cell apoptosis and related proteins Bcl-2 and Bax in the rats with focal cerebral ischemia reperfusion(I/R) injury.Methods Seventy-two healthy SPF male Sprague-Dawley rats were selected and then randomly divided into the control group(Ⅰ:without any treatment),sham operation group (Ⅱ),cerebral IRI group (Ⅲ) and hydrogen gas treatment group (Ⅳ),18 cases in each group.Focal cerebral ischemia reperfusion injury (IRI) model was induced by using the suture-occluded method.The neurological deficits score (NDS) was assessed at 24 h after cerebral reperfusion in four groups.The cerebral infarction severity and size were detected by TTC staining and neuronal apoptosis of brain cortex were tested by TUNEL technique.The apoptosis index (AI) was calculated.Then the expressions of GRP78,Caspase-12,Bcl-2 and Bax were assessed by Western blot and immunohistochemistry.Results As compared with the group Ⅰ and Ⅱ,NDS score,cerebral infarction size,AI and the expressions of GRP78,Caspase-12 and Bax in cerebral cortex in the group Ⅲl and Ⅳ were significantly increased,while the expression of Bcl-2 in cerebral cortex was markedly decreased(P＜0.05);compared with the group Ⅲ,NDS score,brain infarction size,AI and the expression of Caspase-12 and Bax in cerebral cortex in the group Ⅳ were markedly decreased,while the expressions of GRP78 and Bcl-2 were dramatically increased (P＜0.05).Conclusion Inhaling high concentration of hydrogen gas has a certain protective effect on cerebral IRI in rats through increasing endoplasmic reticulum GRP78 protein expression after IRI and inhibiting Caspase-12 activation,thus inhibiting ERS and promoting the repair function of endoplasmic reticulum.

Objective To study the effect of Hydrogen on ischemia/reperfusion(I/R)-induced cardiocyte apoptosis and apoptosis related proteins expression in diabetic rats .Methods Diabetes mellitus was induced by intraperitoneal injection of streptozotocin (STZ) ,then feed 4 weeks before build ischemia/reperfusion model .60 SD male rats ,weight during 300-350 g ,were randomly divid-ed to six groups :non-diabetic sham-operated group (NS) ,non-diabetic I/R group(NI/R) ,non-diabetic hydrogen treated group (NH) ,diabetic sham-operated group(DS) ,diabetic I/R group(DI/R)and diabetic hydrogen treated group(DH) ,each group has 10 rats .The cardiac muscle I/R model was made by 30 min occlusion of the left anterior descending coronary artery and 2 h reperfu-sion .The rats in Hydrogen group were treated with 5 mL/kg hydrogen by intraperitoneal administration at the beginning of reper-fusion .The apoptosis index (AI) was calculated by TUNEL .The positive expressions of Bcl-2 ,Bax ,Caspase-3 in cardiomyocytes were respectively detected by immunohistochemistry .Results The apoptotic rates of cardiomyocytes and the positive expressions of Bcl-2 ,Bax ,Caspase-3 were significantly increased(P<0 .01) ,Compared with I/R group ,the apoptotic rates of cardiomyocytes in hydrogen treatment group were obviously decreased(P<0 .01) ,and the positive expressions of Bcl-2 were increased(P<0 .01) ,at the same time ,the positive expressions of Bax ,Caspase-3 were decreased(P<0 .01) .Conclusion Hydrogen inject by intraperitoneal method on myocardial ischemia-reperfusion injury of diabetic rats has a protective effect ;Its mechanism may be related to its inhibi-ting myocardial apoptosis by advanced the Bcl-2 protein expression and reduced the Bax 、Caspase-3 protein expression .

Objective To summarize the correlation between cytotoxic T lymphocyte associated antigen-4 (CTLA-4) promoter-49 A/G polymorphism and Graves' disease (GD)in China.Methods The publications before Sep.30,2011 about the correlation between CTLA-4 promoter-49 A/G polymorphism and GD in China were collected by searching CNKI,CBM,and Wanfang database.Meta-analysis was performed to check the difference of genotypes such as AG,GG,GG + AG,and G allele between GD group and the control group.Results A total of 6 studies involving 570 cases in GD group and 486 cases in the control group were included in the meta-analysis.According to the heterogeneity of these studies,meta-analysis was performed to analyze the correlation between AG,GG,GG + AG genotype,G allele and GD by fixed effects model.The pooled OR value(95％ CI) of various studies was 1.97 (1.37-2.82),3.66 (2.42-5.53),2.55 (1.82-3.58),and 1.99 (1.49-2.65).Conclusion The meta-analysis demonstrates that CTLA-4 promoter-49 A/G polymorphism increases the possibility of getting GD in Chinese.

Objective To investigate the effect of high concentration hydrogen gas on neurons in the rat hippocampus CA1 region during global cerebral ischemic/reperfusion injury (GCIR) Methods Four-vessel occlusion was used to establish rat model with GCIR injury. One hundred and five healthy male Sprague-Dawley rats were randomly divided into sham operation group(SH group, n = 15), model group(4-VO group, n = 45) and treatment group(4-VO+H2 group,n = 45). After 72 h and 9 d reperfusion, hippocampal CA1 region pyramidal neurons in every group were detected with Nissle staining , immunohistochemical neuron-specific nuclear protein (NeuN), specific protein antibody microglial cells (Iba1) staining and the relationship of position between neurons and microglia was observed through fluorescence double staining. We used Morris water maze to test the space orientation ability and the learning and memory ability in rats after 9 d reperfusion. Results Compared with those of 4-VO group,the neurons of hippocampus CA1 region were closer to normal in 72 h and 9 d in 4-VO+H2 group and neuron form and the number of neuron survival were increased significantly (P < 0.05);immunohistochemical staining showed that the number of neuron survival in 4-VO+H 2 group was obviously higher than that in 4-VO group (P < 0.05) and the number of microglia in 4-VO group was obviously higher than that in 4-VO+H2 group (P < 0.05). Water maze experiment showed that the swimming time in quadrant Ⅳ in 4-VO+H2 group was longer than that in 4-VO group (P < 0.05). Conclusion Inhalation of high concentration hydrogen gas has prominent protective effect on neurons of rat hippocampal CA1 region during reperfusion. The mechanism may be related with inhibiting the microglia excitation and activation during GCIR.