Objective To study the expressions of poly(ADP-ribose) polymerase (PARP) and apoptosis-inducing factor (AIF) in the hippocampal CA1 region of rats after focal cerebral ischemia reperfusion injury,and elucidate the neuroprotective effect ofcilostazol pretreatment through PARP/AIF pathway.Methods One hundred and thirty-five male SD rats were randomly divided into three groups (n=45) for experiments:sham-operated group,ischemia/reperfusion (I/R) vehicle group and cilostazol pretreatment group.Embolization thread was inserted into all rats except those in the sham-operated group for 2 hours to establish middle cerebral artery occlusion models; and then,reperfusion for 6,24,72 hours was performed,and rats of each group were sub-divided into three groups according to reperfusion times (n=l 5).Cilostazol pretreatment group was given cilostazol (30 mg/kg,twice orally) before surgery.The apoptosis cells of the hippocampus were detected by Terminal deoxynucleotidyl Transferasemediated dUTP nick end-labeling (TUNEL).Western blotting was used to test the dynamic changes of AIF and polyadenosine diphosphate ribose (PAR) levels at different times.The mRNA expression of AIF was assessed by real time-PCR.Results There appeared subsequent translocation of AIF after focal cerebral ischemia reperfusion injury in rats.As compared with those in the sham-operated group,the apoptotic cells statistically increased; AIF and PAR contents and AIF mRNA expression significantly increased in the I/R vehicle group reaching its peak level at 24 h ofreperfusion (P＜0.05).As compared with of the I/R model subgroups,the cilostazol pretreatment subgroups had significantly decreased AIF and PAR contents,AIF mRNA expression and number of apoptosis cells (P＜0.05); 24 h of reperfusion subgroup had the most obvious effect (P＜0.05).Conclusion Cilostazol pretreatment in rats after ischemia-reperfusion injury has certain neroprotective effect,whose resistance roles in nerve cell apoptosis may be implemented by inhibiting excessive PARP activation and AIF translocation caused by cerebral ischemia injury.

Objective To investigate the regulative effect of exogenous Angiotcnsin-(1-7) [Ang(1-7)] on nuclear factor-κB (NF-κB) and its downstream proinflammatory cytokines in rats after cerebral ischemic-reperfusion injuries.Methods Forty-two male SD rats were randomly divided into sham-operated group,control group and Ang-(1-7) treatment group (n=14); cerebral ischemia in control group and Ang-(1-7) treatment group was induced by transient middle cerebral artery occlusion (MCAO).The rats in the sham-operated group and control group were infused with artificial cerebrospinal fluid (aCSF,0.5 μL/h) while rats in the Ang-(1-7) treatment group were with Ang-(1-7)(100 pmol,0.5 μL/h)into the lateral ventricle by implanted osmotic minipumps following reperfusion.At 24 h after reperfusion,all rats were sacrificed to detect the expression of NF-κB p65 subunits in cell nuclei of the ischemic cortex by Western blotting.The spatial distribution of NF-κB p65 subunits in ischemic cerebral tissues was detected by immuno-histochemical assay.The concentrations of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) in serum were detected by ELISA.Results A significant reduction of NF-κB p65 expression in cell nuclei by 46％ in the Ang-(1-7) treatment group was noted as compared with that in control group (P＜0.05).The nuclear translocation of NF-κB p65 in the ischemic cortical cells in the Ang-(1-7) treatment group was significantly reduced,and the rate of NF-κB p65 positive cells was decreased by 29％ as compared with that in control group (P＜0.05).Besides,the serum levels of TNF-α (71.603±18.539 pg/mL) and IL-1 β (44.648±10.387 pg/mL) in the Ang-(1-7) treatment group were obviously decreased as compared with those in the control group (104.763±24.412 pg/mL,64.787±14.441 pg/mL,P＜0.05).Conclusion Exogenous administration of Ang-(1-7) could attenuate inflammatory reaction following cerebral ischemia,perhaps by interacting with Mas receptor or through the antagonism against the pro-inflammatory effect of Ang Ⅱ.

Objective To study the role of inflammation in white matter damage of spontaneously hypertensive rats (SHRs) through observing the pathology changes of tissues after white matter damage and detecting the levels of inflammation-related indicators.Methods Eighteen 40-week-old male SHRs were chosen as experimental group,and seven male Wistar-Kyoto (WKY) rats were used as control group.The animal brain tissues were taken for hematoxylin-eosin staining (HE staining) and immunohistochemical staining to observe the pathological changes,and the levels of myelin basic protein (MBP),neurofilament (NF) and glial fibrillary acidic protein (GFAP).Real-time PCR was employed to detect toll-like receptor 4 (TLR-4),monocyte chemotactic protein 1 (MCP-1) and vascular cell adhesion molecule-1 (VCAM-1) expression levels in the white matter tissues.Results The white matter of 40-week-old SHRs was apparently injured.HE staining displayed sponge-like changes in the white matter and immunohistochemical staining showed astrocyte activation,reduced number of axonal and demyelination in the white matter.As compared with those in the WKY rats,TLR-4,MCP-1 and VCAM-1 mRNA expressions in SHR white matter were significantly increased (P＜0.05); and TLR-4,MCP-1 and VCAM-1 expression levels in SHRs were positively related to the degree of white matter damage.Conclusion The white matter damage in 40-week-old SHRs is similar to that of LA;inflammation is involved in the pathophysiological process of white matter damage,being one of induced factors of white matter injury.

Objective To investigate an appropriate activation method and culture system in vitro of rabbit parthenogenetic embryos to make an experimental foundation for therapeutic cloning.Methods Mature rabbit oocytes with polar body 1 (PB 1) were divided randomly into four groups and accepted,respectively,parthenogenetic activation with 1.2 kV/cm,1.6 kV/cm and 2.0 kV/cm electricity stimulation or 5 μg/mL ionomycin (ION) chemical treatment; the effects of these different treatments on activation efficiency were compared.The activated oocytes were cultured,respectively,in M199 supplemented with various concentrations of leukaemia inhibitory factor (LIF,0,10,20,40 and 80 ng/mL) or insulin+transferrin+sodium selenite (ITS,0×,1 × and 2×); and then,the rate of cleaved oocytes,percentage of morulas/blastocysts and total number of blastocysts were compared between each two groups.Results (1) Different treatments on the oocytes showed significantly different activation effects.The survival rate of oocytes activated in 5 μg/mL ION for 4 min were significantly higher than that of ones stimulated by 2.0 kV/cm electricity treatment (P=0.016).The percentages of cleaved oocytes,developing 8-16 cells and morulas/blastocysts in the ION treatment group were also significantly higher as compared with those in the 1.2 kV/cm electric treatment group (P=0.000,P=0.002 andP=0.026,respectively).(2) Significant difference in the percentage of morulas/blastocysts was noted between 20 ng/mL and 80 ng/mL of LIF supplementation (P=0.003); in addition,the total number ofblastocysts in the medium supplemented with 20 ng/mL LIF was significantly different to those with 40 ng/mL and 80 ng/mL,respectively (P=0.011,P=0.002); In experiment of ITS supplementation,significant difference in the percentage ofmorulas/blastocysts was shown between 1 × and 2× ITS supplementation (P=0.003); in addition,the total number of blastocysts in the medium supplemented with 0× or 1 × ITS (166 and 147 cells) was significantly different as compared with that with 2× ITS (78 cells,P=0.015,P=0.044).Conclusion The parthenogenesis activation of the New Zealand rabbit's oocytes treated with ION+6-DMAP shows a higher rate of cleaved oocytes and blastocysts than that with electricity stimulation; the subsequent development of parthenogenetic embryos in vitro could be significantly improved by supplementing with 20 ng/mL LIF and 1× ITS.

Objective To assess the functional changes of the hind paw after SD rats being cut offthe L5 spinal nerve by CatWalk gait analysis.Methods Eleven male SD rats were chosen; the right side of these rats,as the experimental side,was cut the ventral root and dorsal root of L5 spinal nerve,while the left side was without any operation.One day after the operation,whether the models were successfully constructed was verified by tracer experiment with wheat germ agglutinin horseradish peroxidase (WGA-HRP) in 3 rats; 1 day before the operation and 3 months after the operation,the changes of such parameters as general parameters,individual paw parameters,pain-related parameters and coordination-related parameters were assessed by using CatWalk gait analysis.Results The WGA-HRP-labeled motoneurons and fibers were not found in posterior horn and anterior horn of the experimental side of L5 spinal gray matter,but both located in the left side,indicating that the models were constructed successfully.General parameters,individual paw parameters and pain-related gait parameters could be recorded 3 months after the operation,but no statistically significant difference was noted as compared with those 1 day before the operation; however,significant decrease of values of regularity index (RI) and coupling diagonal right after-left front in rats 3 months after the operation was noted as compared with those 1 day before the operation (P＜0.05).Conclusion Only a part of coordination-related gait parameters (the lower limbs) is influenced after being cut L5 spinal nerve;therefore,L5 spinal nerve can be used for constructing of a bladder reflex arc.

Objective To investigate the related anatomical parameters of single-nostril transsphenoidal approach in adults.Methods Two hundred and ninty-four adults,performed head-neck CTA in our hospital from October 2011 to January 2012,were prospectively collected; and their CT data were collected,and the following anatomical parameters were measured through the multi-planar reconstruction of the CTA images:the angle of sella lowest point-anterior nasal spine root-the bottom of nasal cavity (∠ 1),the angle of the connection of Maxillary central incisors bottom to auditory canal crossing with the bottom of nasal cavity(∠ 2),the angle of tuberculum sellae-anterior nasal spine root-the bottom of nasal cavity(∠3),the angle ofdorsum sellae-anterior nasal spine root-the bottom of nasal cavity (∠4),and the angles of right / left sphenoid sinus mouth-maxillary ridge-the bottom of nasal cavity(∠5 / ∠6).Results ∠1(29.7±3.1°),∠2(29.5±3.1°),∠3(33.6±3.8°),∠4(28.8±3.6°),∠5(33.9±4.2°) and ∠6(33.6±4.5°) were calculated; significant differences were noted between each two angles (P＜0.05); ∠ 1 and ∠2 had a positive linear correlation (r=0.78,P＜0.05).Conclusion Preoperative multi-planar reconstruction images of CT facilitates safe opening of the anterior wall of spheniodal sinus and the sellar floor,and the operative position refers to the orientation that the virtual plane of Maxillary central incisors bottom to bilateral auditory canal erects the ground.

Objective To observe the influence of combination therapy ofprobucol,aspirin and statins drugs (PAS) on levels of serum oxidized low-density lipoproteins (ox-LDL),pregnancy-associated plasma protein A(PAPP-A) and marix metalloproteinase-3(MMP-3) and resolution of carotid vulnerable plaque in patients with acute cerebral infarction (ACI).Methods One hundred and thirty-five patients with ACI,admitted to our hospital from September 2007 to July 2010,were chosen in our study; according to the results of carotid artery ultrasound,these patients were divided into carotid stable plaque group (n=45) and carotid vulnerable plaque group (n=90).Stable plaque group was considered as control group; 90 patients with carotid vulnerable plaque were randomly subdivided into aspirin and statins drugs (AS) group (n=45,aspirin 100 mg/d,atorvastatin 20 mg/d,orally) and PAS group (n=45,aspirin 100 mg/d,atorvastatin 20 mg/d,probucol 0.25 g/time,2 times/day,orally).Levels ofOx-LDL,PAPP-A and MMP-3 were detected in all patients before treatment and four weeks after drug therapy.The intima-media thickness,plaque area and echogenicity of carotid plaques were evaluated by Doppler ultrasonography during a 12-month follow-up period.Results Before treatment,serum ox-LDL,PAPP-A and MMP-3 levels in AS group and PAS group were significantly higher than those in the control group (P＜0.05); no significant differences of ox-LDL,PAPP-A and MMP-3 levels were noted between AS and PAS groups (P＞0.05).Four weeks after treatment,levels of serum ox-LDL,PAPP-A and MMP-3 in PAS group were significantly lower than those in AS and control groups (P＜0.05); the decrease of levels of ox-LDL,PAPP-A and MMP-3 in PAS group was obviously higher than that in AS and control groups (P＜0.05).Twelve months after treatment,significant decrease of plaque area and intima-media thickness (IMT) was noted in the AS and PAS groups (P＜0.05); the IMT and plaque area in PAS group were significantly smaller than those in AS group (P＜0.05); obvious decrease of echogenicity of carotid plaques in PAS group was noted as compared with that in AS group (P＜0.05).Conclusion Combination therapy of PAS might have strong anti-oxidant function and lipid-lowering effect,which could reverse and stabilize the atherosclerosis plaque.

Objective To investigate the efficacy of erythropoietin (EPO) on cerebral ischemia of patients after intracranial aneurysm surgery.Methods Sixty patients with cerebral ischemia after operation of intracranial aneurysm,admitted to our hospital from May 2006 to August 2010,were randomly divided into control and treatment groups (n=30).Control group received conventional treatment,while the other group,on the basis of conventional therapy,was hypodermically given EPO at a dosage of 3000 IU for a consecutive 3 days.U.S.National Institutes of Health Stroke Scale (NIHSS) was used to score the patients on the admission day,1,2,3,4 and 5 weeks after EPO treatment; and diffusion-weighted imaging (DWI) of MRI was used in determining the changes of cerebral ischemic area on the ischemic day and l,2,3,4 and 5 weeks after ischemic.Results NIHSS scores of the EPO group (9.4±1.9,11.3±2.3,8.7±1.7) at 3,4 and 5 weeks after treatment were significantly higher as compared with those in the control group (10.8±2.2,7.9±1.6,10.1±2.3) (P＜0.05).Similarly,3,4 and 5 weeks after treatment,the reducing percentage of cerebral ischemia area in the EPO group (21.6±4.1,27.5±5.3,32.8±5.5)％ was larger than that in the control group (16.8±3.1,18.9±3.3,19.5±3.4)％ (P＜0.05).Conclusion EPO may play an effective role in cerebral ischemia of patients after intracranial aneurysm operation.

Objective To investigate the relationships between the risk factors (the levels of blood glucose and blood fat) and the severity of leukoaraiosis (LA) in patients with type 2 diabetes.Methods The risk factors (including levels of plasma HbA1c,fasting blood glucose,2 hours blood glucose after meals,triglyceride,total cholesterol,high density lipoprotein,and low density lipoprotein) were measured and the head magnetic resonance were used in the selected patients who were diagnosed as having subcortical vascular dementia and type 2 diabetes (n=106).According to the Fazekas's classification of signals of deep white matter hyperintense (DWMH) and periventricular hyperintense (PVH),the severity of LA was graded as grade 1,2 and 3 by its appearance under MRI.LA scores were obtained by its appearance under MRI according to the semiquantitative way; the influencing factors of LA scores were analyzed using multiple linear regression.The clinical features of patients with different plasma HbAlc levels were compared; the relations of signals of DWMH and PVH with other risk factors were compared; and the clinical features of patients with different signals of DWMH and PVH were compared too.Results LA scores were positively related to HbAlc level and age (r=0.457,P=0.000;r=0.400,P=0.000).The signals of DWMH and PVH were positively related to the HbA1c level and age (r=0.385,P=0.000; r=0.361,P=0.000; r=0.458,P=0.000; r=0.364,P=0.000).Statistical significance of stroke history in different signals of DWMH and PVH was noted (P＜0.05).After adjusting the age,signals of DWMH and PVH were positively correlated to the HbA1c level (r=0.253,P=0.009; r=0.318,P=0.001).Conclusion In the patients with type 2 diabetes,the plasma level of HbA 1 c is an important biochemical indicator which reflects the severity of LA (including DWMH and PVH); age and stroke history are relative with the severity of LA,while blood fat level is not correlated to the severity of LA.

Objective To investigate the distribution of blood vessels and the relationship of angiogenesis with hypoxia-inducible factor-1α (HIF-1α) and vascular endothelial growth factor (VEGF) in meningiomas.Methods Fifteen specimens of meningiomas,collected in our hospital during the resection and conformed by pathology from May 2010 to March 2011,were chosen; 4 different tissues from each specimen were taken,including the tail sign,base center,tumor' s center and tumor' s margin; three successive slices were made from each tissue part.Envision immunohistochemistry was employed to detect the expressions of CD34,VEGF and HIF-1α and their different distributions were compared.Results CD34 mainly disturbed in the cell membrane and cytoplasm,showing yellow granules in the dyeing; VEGF located at the cytoplasm of the tumor cells,while HIF-1α mainly at the nucleus.The expressions of CD34,VEGF and HIF-1α in the tail sign and tumor's margin were significantly higher than those in the MVD in the base center and tumor's center (P＜0.05).Positive correlations between expressions of VEGF and MVD,expressions of HIF-1α and MVD,as well as expressions of HIF-1α and VEGF were noted (r=0.960,P=0.040; r=0.964,P=0.036; r=0.998,P=0.002).Conclusion The angiogenesis in the periphery ofmeningiomas is much richer than that in the center; H1F-1α and VEGF may induce and promote the angiogenesis of meningiomas.