Objective To explore the effects of repetitive transcranial magnetic stimulation on behaviors and hippocampus neuronal apoptosis in rats with chronic stress-induced depression.Methods 40 male SD rats of SPF grade were randomly divided into normol control group (n =8) and model preparation group (n =30) after screening.Rats in model preparation group were singly housed and given chronic unpredictable mild stress(CUMS) to build depression model.Excluding unsuccessful modeling rats,the model preparation group was divided into three groups:model group(n=8,without any treatment),rTMS group (n=8,with the intervention of 10 Hz rTMS) and shame group (n=8,simulation of rTMS environment without rTMS stimulus).The changes of behaviors in each group were detected by weight measurement,sucrose consumption test and open-field test.The changes of morphology of hippocampal neurons were detected by Nissl's staining.The changes of Bax in hippocampal neuron were detected by Immunohistochemical staining.Results (1) Behavioral results showed stress for 21 d could make rat behavior scores decrease significantly(all P＜0.05),and rTMS intervention could significantly improve their behavior scores (all P＜0.01).Compared with model group,the weight reduction rate (0.32±0.05)％,the score of sucrose consumption test(7.03 ± 1.02) and the score of open field test(8212.41 ± 1416.15,8.75 ± 1.58) in rTMS group was higher(P ＜0.01).(2) Nissl staining showed stress for 21 d could make the number of hippocampal CA3 neurons was reduced,cell morphology was poor,and the number of Nissl bodies was reduced.rTMS intervention could increase the number of hippocampal CA3 neurons,cell morphology was integral and the number of Nissl bodies was increased.(3) Immunohistochemistry results showed stress for 21 d could cause the number of Bax cell were significantly increased(P＜0.01),and rTMS intervention can make the number of Bax cell were significantly lower(P＜0.01).Conclusion rTMS intervention improves the depressive behavior in chronic stress depression model rats and inhibits the apoptosis,which might work through inhibition of neuron apoptosis and decline of Bax expression in hippoeampal neurons.

Objective To explore the effect of repetitive transcranial magnetic stimulation (rTMS) on behaviors and hippocampal glucocorticoid receptor (GR) protein expression in chronic stress depression model rats and the possible antidepressant mechanism of rTMS. Method Seventy-five male Sprague-Dawley rats were randomly divided into the blank control group (n=15) and the stress-induced group (n=60). Singly housing and chronic unpredictable mild stress (CUMS) were used to induce the depression model in stress-induced group. Forty-five CUMS rats were selected and ran?domly divided into rTMS group (receiving 10 Hz rTMS intervention for 3 weeks), sham group (receiving pseudo rTMS treatments for 3 weeks) and depression group (with no further treatment). Body weight measurements and performance in the sucrose consumption and forced swimming test (FST) were evaluated before modeling, after modeling and after inter?vention. The GR protein and GR mRNA expression level in the hippocampus were examined after intervention. Results Compared with control group, the body weight growth rate and the sugar water preference were significantly lower in stress-induced group (P<0.01), and the immobility time of FST was significantly longer (P<0.01). After the 3-week rTMS intervention, the body weight growth rate and the sugar water preference in rTMS group, which were insignificantly differ?ent from control group (P>0.05), were higher than those in sham group and depression group (P<0.01). The immobility times of FST in rTMS group and control group were shorter than sham group and depression group (P<0.01). Compared with rTMS group and control group, GR and GR mRNA expression levels in the hippocampus were significantly reduced in sham group and depression group (P<0.01). Conclusion rTMS can improve depression behavior of CUMS rats, which may be associated with upregulation of GR expression in the hippocampus.

The purpose of this study was to evaluate the effect of adenosine A2A receptor antagonist ZM241385 on amygdala-kindled seizures and its roles in epileptogenesis. Electrodes were implanted into the right amygdala of male adult Wistar rats. Kindling was accomplished by using stimulus strength of 500 μA applied daily to the amygdala until 10 consecutive stage 5 seizues were induced. Then effect of ZM241385 was studied in fully kindled rats after intracerebroventricular administration of the drug. In addition, the effect on kindling progression was evaluated through ZM241385 injection before daily stimulation. In all experiments, behavioral changes in the rats in response to ZM241385 were monitored closely. The results showed that, in fully amygdala-kindled rats, ZM241385 (0.001-0.1 nmol/L) decreased afterdischage duration (ADD), motor seizure duration (MSD), stage 5 duration (S5D) and seizure duration (SD), but only the effect on ADD was dose-dependent. The doses of 0.001-0.1 nmol/L had no influence on stage 4 latency (S4L) and seizure stage (SS). The dosages of 0.0001 and 1 nmol/L of ZM241385 did not exert any effect on all seizure parameters. In contrast to the results in fully amygdala-kindled rats, ZM241385 (0.001-0.1 nmol/L) had minimal or no effects on the progression of amygdala-kindled seizures. We are led to the conclusion that although ZM241385 had no influence on the progression of amygdala-kindled seizures, it had potent anti-convulsant profile and little adverse effects at the dosage of 0.001-0.1 nmol/L, suggesting that the agent is effective against the amygdala-kindled seizures.

Objective To investigate the dynamic changes of adenosine receptors, A1 (A1R) and A2a (A2aR) in the brain from the acute to chronic phase after kindling and to explore the correlation between seizure and expression level of A1R and A2aR. MethodsRats were randomly selected into the testing model, reference and normal control groups.Testing rats were kindled by lithium choride-pilocarpine, reference rats were treated with saline, and no treatment was given in normal control group. The dynamic expression of A1R and A2aR were detected by RT-PCR, immunofluorescence staining and Western blot at time-points of 24 hours, 1 month and 6 months post-kindling. Results In the acute phase of 24 hours after kindling, the A1R expression level (mRNA level was (1. 1483 ±0. 1182); Western blot result was ( 0. 7872± 0. 0621 ) ; immunofluorescence staining count was ( 76. 17 ± 4. 62 )/HP) was increased and A2aR (mRNA level was (0. 8338±0. 0572) ; Western blot result was (0. 2098 ±0. 0257) ; immunofluorescence staining count was (43. 83 ± 5. 12 )/HP) was decreased.The results showed statistically difference compared with the reference and normal groups (P< 0. 05 ). In the later chronic phase of 1 month and 6 months after kindling, the expression level of A1R was decreased and A2aR was increased. These data revealed statistically significant difference (P <0. 01 ). Furthermore, the comparison of the results in 1 month and 6 months after kindling found that the expression of AIR was lower in 6 months (mRNA level was (0. 5682 ±0. 0443) ; Western blot result was (0. 7749 ±0. 0262) ; immunofluorescence staining count was (38. 50 ±4. 81 )/HP) than in 1 month and that of A2aR was higher in 6 months (mRNA level was (1. 2169±0. 0332) ; Western blot result was (0. 7080 ±0. 0371 ); immunofluorescence staining count was (114. 50 ± 4. 04)/HP). The differences were statistical significant (t = - 19. 02--13.28, P < 0. 05). ConclusionsThe expressions of A1R and A2AR during and after kindling presents a bidirectional change. In the acute phas the expression of AR is regulated to suppress seizures. While in the chronic phase, the repeated seizures result in the change of A1R and A2aR expression in the opposite direction. This mechanism plays an important role in refractory seizures.

Objective To study the recovery mechanism of dysphagic patients after stroke using functional magnetic resonanee imaging(fMRI). Methods Thirteen patients with dysphagia caused by unilateral cortical or subcortical lesions were recruited into a dysphagia group,and eight age-matched healthy volunteers were recruited as controls.Both grouDs performed experimental volitional swallowing tasks during fMRI studies.All patients of the dys-phagia group received rehabilitation treatment targeting dysphagia.Of the 13 dysphagia patients,7 reached almost complete recovery and were identified as recovered in follow-up fMRI studies.A 3.0 T MR scanner and echo planar imaging(EPI)T_2 WI sequence were employed to obtain the fMRI data.SPM2 software was used for post-processing of the fMRI data and displaying activated brain maps.Lateral index(LI)was calculated as LI:(C-1)/(C+I).Paired t tests were used to compare activated brain volume before and after complete recovery. Results Consistent activation of the bilateral primary sensorimotor cortex,anterior cingulated gyrus and the bilateral insular cortex were observed in the control group. Activation of the pons,medulla,left cerebellum,left prefrontal area,right occipital area and the left insular cortex were observed in the dysphagia group.Activation was observed in the bilateral primary sensorimotor cortex.bilateral prefrontal area,bilateral superior temporal gyrus,left insular cortex,bilateral frontal o-pereulum and anterior cingulated gyrus in the recovered patients.The total activated volume before recovery in the ip-silesional hemisDhere was significantly less compared with the contralesional hemisphere in the dysphagia group.In the recovered patients,both the activated brain volume of the ipsilesional hemisphere and value of LI were significant-ly larger than those at the initial examination.Conclusions Decreased activation in the sensorimotor cortex,the in-sular lobe and the cingulate gyms might be causes.of dysphagia.Compensation by the contralesional hemisphere in the early stages and then the restoration of the ipsilesional hemisphere after recovery may be mechanisms of dysphagia recovery in stroke patients.

Objective To observe the effect of repetitive transcranial magnetic stimulation (rTMS) on behavior in response to chronic but unpredictable mild stress and explore potential neuroendocrine mechanisms.Methods Forty adult SD male rats were randomly divided into a control group (n =8) and a model preparation group (n=32).The control group was given normal care while a model of depression was induced in the model preparation group through giving an unpredictable mild stimulus (CUMS).The depressive rats were randomly divided into a model group,an rTMS group and a sham rTMS group (8 cases in each group).The rTMS group and sham rTMS groups accepted the rTMS or sham stimulation for 3 weeks.The changes in behavior in each group were quantified using body weight,sucrose consumption and an open field test before and after stimulation.Enzyme-linked immunosorbent assays (Elisas) were conducted to detect plasma adrenocorticotropic hormone (ACTH) and corticosterone (CORT) levels.Reverse-transcription polymerase chain reactions (RT-PCRs) were carried out to allow the detection of mRNA expression in hypothalamus related to levels of adrenocorticotropic hormone releasing hormone (CRH).Results After the modeling there were significant differences between the model preparation group and the control group in terms of weight increase,sucrose consumption and open field test results.After rTMS the rate of weight increase,sucrose consumption and the scores in the open field test of the rTMS group had increased significantly more than in the control group.Elisas showed significantly higher plasma ACTH and CORT levels in the model group as well.The average expression of CRH mRNA in the model group was significantly higher than in either of the other two groups.Conclusions rTMS can relieve depression-like behavior induced by chronic stress,at least in rats.This may be related to a downgrading of the hyperactive functioning of the hypothalamus-pituitary-adrenal axis.

Objective To investigate the polymorphisms of-139 and -336 nucleotides in dendritic cells specific intercellular adhesion molecule-3-grabbing nonintegrin (DC-SIGN) promoter region in context of HIV susceptibility, infection routines and HIV/AIDS progress. Methods Polymorphisms of -139 and -336 nucleotides in DC-SIGN were examined in 160 HIV-positive subjects and 178 healthy controls;the Spearman test was performed to analyze their associations with HIV infection status. Results In 160 HIV-positive subjects, there were 92 (57.5%) with-139C, 68 (42.5%) with-139T, 29 (18.1%) with-336C and 131 (81.9%) with -336T. The frequencies of -139T/C and -336T/C in HIV-positive subjects were similar to those in the healthy controls (χ2 =0. 121 and 1. 754, P ＞0.05 ). No differences were found in the distribution of -139T/C or -336T/C in HIV-positive subjects infected via sex intercourse or intravenous drug (χ2 =0. 435 and 0. 103, P ＞ 0. 05 ). -139C was usually companied with -336C ( r = 0. 359, P ＜ 0.01 ).-139T (27.9%) were more frequently presented in patients with CD4 +T cells ≤50 cells/μL than -139C( 23.0%, χ2 = 4.055, P ＜ 0.05 ). -139T/C and -336T/C were not related to HIV RNA levels ( t = - 0. 643and - 1. 637, P ＞ 0.05). Conclusions Genotype -139C in DC-SIGN promoter region usually coexist with -336C. Polymorphisms of -139 and -336 are not related to HIV susceptibilities or HIV infection routes.-139T genotype may be related to serious depletion on CD4 + T cells.

Objective To investigate the influences of grape seed proanthocyanidin extract (GSPE) on cardiovascular system, nervous system and respiratory system of experimental animals, and provide general pharmacological data for further research and application. Methods The influences of GSPE on blood pressure, heart rate, electrocardiogram, breathing frequency and tidal volume in anesthetic dogs after duodenal administration were observed, the impacts on spontaneous activity, coordinated motion, and the sleep situation with threshold dose and subthreshold dose of pentobarbital sodium in mice after intragastric administration were observed. Results GSPE showed no side effects on blood pressure, heart rate, electrocardiogram, breathing frequency and tidal volume in anesthetic dogs at the dosage of 857.00, 214.29, 42.86 mg/kg (P>0.05). At the dosage of 428.57, 214.29, 42.86 mg/kg, GSPE had no obvious influence on spontaneous activities and coordinated movements in mice (P>0.05). GSPE did not evidently change the number of sleeping animals, the sleep latency and the sleeping duration with subthreshold dose and threshold dose of pentobarbital sodium (P>0.05). Conclusion GSPE has no evident adverse effects on central nervous system, cardiovascular system and respiratory system in animals.

Objective To clinically analyze the frequent stroke localizations of moyamoya disease in order to improve our cognition toward it and reduce missed diagnosis. Methods All 32 patients were prospectively analyzed over the past 10 years in our hospital. Results The ratio of female to male was 1.28 and their age of onset ranged from 7 to 47 years old. Mean age of 5 ischemic stroke patients (15.6%) was 24 and mean age of 21 haemorrhagic stroke patients (65.6%) was 33 while mean age of 6 mixed type stroke patients (18. 8% ) was 32. The frequent ischemic stroke localizations were frontoparietal lobe (60%),temporo-occipital lobe (20%), and periventricular zone (20%). The frequent haemorrhagic stroke iocalizations were periventricular zone (42.8%), ventricle (39.2%), temporo-occipital lobe (10. 8%) and subarachnoid space (7.2%). No cerebellum and brain stem stroke occurred. Conclusion Adult patients usually develop intracranial hemorrhage. Moyamoya disease should be considered severely when adult patients develop periventricular and ventricular intracranial hemorrhage, frontoparietal cerebral infarction or adolescent patients develop ischemic stroke, especially companied with epilepsy.

BACKGROUND: It has been reported that in China, human bone marrow mesenchymal stem ceils are mostly harvested from adults. Studies on bone marrow mesenchymal stem cells in children are few.OBJECTIVE: To isolate and expand bone marrow mesenchymal stem cells from children, and to analyze the biological characteristics of bone marrow mesenchymal stem cells and their potential of differentiating into osteoblasts, adipocytes and neural like cells.DESIGN: Observational comparative study.SETTING: Tongji Medical College, Huazhong University of Science and Technology.MATERIALS: Experiments were performed at the Laboratory of Department of Orthopaedics of Wuhan Tongji Hospital from March to September 2006. Bone marrow mesenchymal stem cells were collected from one boy patient and two girl patients aged 5-8 years, who received pelvis osteotomy for dysplasia of the hip joint. The experimental procedures were approved by the Hospital Ethics Committee and family members of all children patients singed the informed consent.Dexamethasone, vitamin C, β-sodium glycerophosphate, 3-1sobutyl-1-methylxanthine, insulin, indometacin and butylated hydroxyanisole were bought from Sigma Company. Dimethyl sulphoxide was purchased from Amersco Company.METHODS: Bone marrow mesenchymal stem cells were cultured from mononuclear cells isolated over a Percoll gradient.Bone marrow mesenchymal stem cells were observed under an inverted phase contrast microscope. Bone marrow mesenchymal stem cells could differentiate into osteoblasts, adipocytes and neural like cells with osteoblast inductor (β-sodium glycerophosphate, dexamethasone, vitamin C), lipoblast inductor (dexamethasone, 3-isobutyl-1-methylxanthine,bovine insulin, indometacin) and serum-free medium inductor (dimethyl sulphoxide, butylated hydroxyanisole) respectively.Osteoblast marker (alkaline phosphatase, osteocalcin mRNA, calcium node), adipocyte marker (lipid droplet, PPAR γ-2mRNA) and neural ceil-like marker (nissl body, neuron specific enolase, neurofilament protein) were respectively determined by the immunohistochemical method, polymerase chain reaction and immunocytochemical method.MAIN OUTCOME MEASURES: ①Appearance and proliferation of bone marrow mesenchymal stem ceils from children,and ②determination results of osteoblast, adipocyte and neural cell markers.RESULTS: ①Children bone marrow mesenchymal stem cells could easily adhere to the wall, appeared fusiform, had high reproductive activity and arranged vortically after fusing. ②Appearance of bone marrow mesenchymal stem cells changed after receiving inductor. Osteoblast marker, adipocyte marker and neural cell-like marker were positive after chemical staining, polumerase chain reaction and immunocyte staining.CONCLUSION: Children bone marrow mesenchymai stem cells show stable proliferation, passage and multi-direction differentiation towards osteoblasts, adipocytes and neural like cells.