Objective To investigate the effect of argenin-glycin-aspartic acid (RGD) peptides on islet viability and function. Methods Rats were randomly divided into 2 groups: 1640 group and RGD group. Ficoll was used to purify islet in a discontinuous-density-gradient way. Islet concentration is 27% ,25% ,23% ,20.5% and 11% respectively. Acridine orange/ethidium bromide(AO/EB) fluorescent staining method was adopted to observe the effect of RGD peptides on islet viability. Radioimmunoassay was adopted to detect insulin level and measure insulin secretion index (SI). Caspase-9 cells and phospho-Akt 473-positive cells were determined by flow cytometry to investigate the influence of RGD peptides on caspase-9 activity and Akt. Results 600-700 IEQ islet was extracted from each rat by Ficoll purification through modified gradient centrifugation. AO/EB stain showed islet survival rate was more than 95% immediately after separation. Islets cultured in the medium of RPMI-1640 showed SI of 1.64 ±0.28 after 1 week, while islets cultured in the medium of RPMI-1640 containing RGD peptides showed SI of 2.28 ± 0.16 (P ＜ 0.05 ). Flow cytometry showed the level of activated caspase-9 was ( 22.66 ± 3.56 ) % if islet was cultured in RPMI-1640 alone for 1 week while the level was( 10.54 ± 1.96) % if islet was cultured in RPMI-1640 containing RGD peptides. There was statistical difference ( P ＜ 0. 05 ). The effect of RGD on Akt phosphorylation was also detected. Akt phosphorylation proportion was (31.47 ±4.08)% 1 week after cultured in RPMI-1640 while the value was(61. 054 ±6.03)% if cultured in RPMI-1640 containing RGD peptides. The difference had statistical significance (P ＜ 0. 05). Conclusion RGD peptides could inhibit apoptosis through the phosphorylation of Akt/PKB.

Objective To evaluate the protective role of bone marrow stromal cells (BMSCs) to lactacystin-Parkinson's disease (PD) rats.Methods BMSCs were obtained from adult male SD rats and were expanded, isolated and purified. BMSCs were harvested from 4 to 6 passage and used for transplantation. PD rats that expressed Lewy body and presented lateral rotation were made by injecting 8?g lactacystin into one side of substantia nigra compact part of rat with stereotaxic technique. The rats were transplanted with BMSCs labelled by Brdu by stereotaxic unilateral injection into striatum and transplanted with normal saline as control. The behavior changes of rats and the expression of BDNF, migration, differentiation of BMSCs were examined.Results 71.43% of the rats displayed significant improvement of contralateral behavior after BMSCs transplantation, however there was no change in control group. BMSCs were found in several areas of the brain including striatum, corpus callosum and contralateral cortex. The implanted BMSCs expressed BDNF but no tyrosine hydroxylase.Conclusion BMSCs may play a neuroprotective role in lactacystin-PD rats by secreting BDNF.

Objective To set up the Parkinson's disease(PD)rat model with pathologic characteristic of Lewy body in nigral cells.Methods SD rats were injected respectively with 8 mg Lactacystin(Lactacystin group),sodium saline(NS group)and 12 mg 6-OHDA(6-OHDA group)by stereotaxic unilateral injection into the pars compacta of substantia nigral.The spontaneous and apomorphine-induced contralateral behaviors of rats were observed.The changes of midbrain histology were viewed by microscope;expression of ?-synuclein and tyrosine hydroxylase(TH)positive cells were investigated by immunohistochemistry.The contents of dopamine and homovanillic acid in striatum were determined.Results Rats of NS group did not display abnormal behavior.The animals treated with Lactacystin developed progressively bradykinesia,hypokinesia,tremor,contralateral head tilting,and displayed apomorphine-induced contralateral rotation behavior;3 weeks later the number of TH positive cells were decreased by 83.29% compared with NS group(P

OBJECTIVE: To investigate the regulatory effect of simvastatin on osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) at middle/late stages by p38MAPK pathway under condition of osteoinductive environment.

Objective To investigate the clinical and electroencephalogram (EEG) features of patients with idiopathic generalized epilepsy in adults with phantom absences.Methods Six patients were referred to the clinic of epilepsy from April,2007 to December,2011.They all had clinical assessment,EEG,or video EEG confirming absences seizure.Results Six patients showed the following similar clinical-EEG features:(1) mild ictal impairment of consciousness associated with generalized 3.0-3.5 Hz spike and slow wave discharges; (2) late-onset generalized tonic-clonic seizures; (3) absence status epilepticus with or without secondary generalized tonic-clonic seizures; (4) generalized discharges were mostly seen in three types in the awaking stage:fragmented discharge (＜4 s),brief discharge (4-10s) and long-time discharge (＞ 10 s).None of the patients had myoclonic jerks or photosensitivity.One patient' s mother had a history of generalized tonic clonic seizures.One patient had a history of children absence epilepsy and one patient had a history of febrile convulsion in the age of 1-3.Conclusion Idiopathic generalized epilepsy with phantom absences has distinct clinical and EEG features and may become a new idiopathic generalized epilepsy syndrome in adults.

Objective To analyze retrospectively the risk factors and predictors of post-encephalitic epilepsy (PEE) and refractory epilepsy in patients with encephalitis.Methods In a hospital based study,the patients with encephalitis were reviewed retrospectively between the January of 1995 and December of 2010.Related factors were evaluated including age,sex,seizure types,neuroimaging,electroencephalogram(EEG) in intermittent period,clinical symptoms,consciousness level,initial seizure and steroid hormone therapy,etc.Results 237 patients with encephalitis were enrolled,whose median age was 26.3 (range 15-57) years old.PEE occurred in 103 (43.46％) patients; and 67 of whom had partial seizure.Significant risk factors for PEE included age (OR =3.72,95％ CI 2.70-5.25,P =0.018),disturbance of consciousness level(OR =5.37,95％ CI 2.43-13.03,P =0.012),cortical lesion in imaging (OR =11.42,95％ CI 5.94-31.27,P =0.000),spike discharges in EEG (OR =18.04,95％ CI 7.30-48.38,P =0.000) and initial seizures in acute phase (OR =32.68,95％ CI 9.62-97.59,P =0.000).The refractory epilepsy occurred in 6t patients.The significant risk factors of refractory PEE included focal seizures(OR =4.09,95％ CI 2.14-9.10,P =0.021),status epilepticus (OR =4.48,95％ CI 1.89-8.07,P =0.017) and poor controlled seizure (OR =6.17,95％ CI 3.52-11.34,P =0.001) during acute phase,multifocal spikes discharge in EEG(OR =5.53,95％ CI 2.91-10.07,P =0.006),cortical lesion in neuroimaging(OR =2.33,95％ CI 1.37-7.72,P =O.028),however,early steroid hormone therapy (OR =2.19,95％ CI 1.11-4.87,P =0.037) and longer time to initial seizure (OR =4.40,95％ CI 3.19-11.62,P =0.014) could significantly reduced the incidence of refractory epilepsy in PEE patients.Conclusion Our data indicated that PEE occur in 43.46％ patients especially in younger patients with disturbance of consciousness level,cortical lesion in imaging,spike discharges in EEG and initial seizures in acute phase.And the risk factors for refractory PEE are also discussed.

Objective To study the effect of proteasome on nigral degeneration and Lewy body formation. Methods By stereotaxic unilateral injection of different doses of lactacystin, a selective proteasome inhibitor, into the substantia nigral pars compacta of rats, the spontaneous and apomorphine induced contralateral behaviors of rats were observed. Nigral degeneration and Lewy body were viewed by HE staining; expressions of ? synuclein and tyrosine hydroxylase in nigral cells were investigated by immunohistochemistry. Contents of dopamine and homovanillic acid were determined by HPLC. Results There were no difference between 0 2 ?g group and control. Animals treated with 2 ?g and 8 ?g lactacystin developed progressively bradykinetic and displayed contralateral head tilting and tremor; apomorphine induced contralateral behavior was notably observed in rats of 8 ?g group; 3 weeks later, nigral degeneration were present in 2 ?g and 8 ?g groups; some of nigral cells contained acidophilic Lewy body with intense immune response to ? synuclein; the number of tyrosine hydroxylase positive cells in 2 ?g and 8 ?g groups were decreased by 68 24% ( P

BACKGROUND:Previous studies have demonstrated that simvastatin that can promote osteogenic differentiation of bone marrow stromal stem cel s in vitro, is likely to be a new osteogenic drug. While it is stil unknown whether there is time-dependent stimulation of simvastatin on the expressions of bone morphogenetic protein 2 and col agen type I. OBJECTIVE:To investigate the expressions of bone morphogenetic protein 2 and col agen type I in rat bone marrow stromal stem cel s in vitro stimulated by simvastatin at different time points. METHODS:Passage 1 bone marrow stromal cel s were divided into control and simvastatin group, fol owed by cultured in osteogenic differetiation medium with or uithout 10-7 mol/L simvastatin. After 7-day intervention, expression of alkaline phosphatase was detected in passage 3 cel s. Passage 4 cel s were divided and cultured as described above, and afterwards, RNA and proteins were extracted at 12 and 36 hours to detect the expressions of bone morphogenetic protein 2 and col agen type I using real-time PCR and western blot assay. RESULTS AND CONCLUSION:Both two groups could express alkaline phosphatase, while the rate of positive cel s significantly increased in the simvastatin group compared with the control group (P<0.05);at 12 and 36 hours after intervention, mRNA expressions of bone morphogenetic protein 2 and col agen type I in the simvastatin group were significantly higher than those in the control group (P<0.05). Besides, western blot assay showed:at both 12 and 36 hours, simvastatin significantly enhanced the expression of bone morphometric protein 2, while the expression of col agen type I significantly increased at 12 hours (P<0.05), but not at 36 hours. In conclusion, simvastatin can promote the expressions of bone morphometric protein 2 and col agen type I in rat bone marrow stromal cel s, with more favorable outcomes after 12-hour treatment.

Objective To investigate the plasma levels of glutamate (Glu) andγ-aminobutyric acid (GABA) in Par?kinson’s disease patients with depression (PDD) and their clinical significance. Methods Plasma levels of Glu and GA?BA were measured in 88 PD patients including 43 PDD patients and 45 PD patients without depression, and 68 healthy controls by using high performance liquid chromatography (HPLC-RF). Depression was assessed in enrolled subjects by using the Hamilton Depression Scale (HAMD). The plasma levels of Glu and GABA were compared among different groups and their associations with HAMD scores were subsequently evaluated by correlation analysis. Results The plas?ma levels of Glu and GABA were significantly lower in PD group(49.81±22.79,249.17±62.57)than in normal control group(149.59±50.08,276.66±85.43)(all P<0.05). In addition, PD patients with depression exhibited significantly low?er plasma levels of Glu and GABA(40.34±15.77 and 233.63±53.56)compared to PD patients without depression(58.86± 24.87 and 264.02±67.39)and healthy controls (all P<0.05). Correlation analysis indicated that HAMD scores were nega?tively associated with plasma levels of Glu ( r=-0.366,P=0.000 ) and GABA ( r=-0.217,P=0.043 ). Conclusion The decrease in plasma levels of Glu and GABA may be implicated in the pathogenesis of depression in PD patients.

Objective To explore the application value of susceptibility weighted imaging(SWI)in measuring brain iron deposition in the diagnosis and the assessment of Parkinson’s disease(PD).Methods Thirty cases with PD underwent head routine magnetic resonance imaging and SWI scanning.The unified PD rating scale(UPDRS)was used to assess the severity of PD.The phase values of substantia nigra (SN),red nucleus(RN),caudate nucleus (CA),globus pallidus(GP)and putamen (PUT)were measured manually on the phase map.The correlation between the phase values of the region of interest(ROI)and the UPDRS scores of PD was analyzed.Results There were no significant differences between the phase values of the less severe body side and those of the more severe body side (SN,P=0.120;RN, P=0.402;CA,P=0.196;GP,P=0.616;PUT,P =0.985).Significant negative correlations were found between the phase values of SN, CA,GP and the UPDRS Ⅲ scores,respectively (SN-UPDRS Ⅲ:r=-0.407,P =0.026;CA-UPDRS Ⅲ:r =-0.424,P =0.02;GP-UPDRS Ⅲ:r=-0.363,P =0.048).Significant negative correlation was found between the phase values of SN and the UPDRSⅤranks (r=-0.373 ,P =0.043 ),while the similar correlation was found between the phase values of CA and the UPDRSⅠ scores (r=-0.367,P =0.046)and the medium negative correlation was found between the phase values of GP and the gait disorder scores of UPDRS Ⅲ(r=-0.41 1,P =0.024).But no correlation was found between the phase values of other ROIs and the UPDRS scores. Conclusion SWI could quantitatively assess the brain iron deposition of the PD patient,which provided reference for clinical diagnosis and assessment of PD.