Objective To simply the method of how to obtain cultures rich in astrocytes from SD rat cerebral cortex which could be utilized in vitro experiments.Methods The neonatal rat cerebral cortex was made into suspension by mechanical dissociation,and then reduced other cells by differential velocity adherent technique,shaking in orbital shaker and passage of cultured cells.After purification,the cultured cells were identified by double immunofluorescence staining and SABC.Results We successfully obtain cultures rich in astrocytes and the proportion of astrocytes were more than 95%.Conclusion The method described above was reliable in obtaining the high purity astrocytes from neonatal rat cerebral cortex and double immunofluorescence staining was more vivid and direct.

OBJECTIVE: To provide information for clinical rational and safe use of albendazole.METHODS: The medical database and network both at home and abroad were searched and the data regarding the research and development,the market information and indications of albendazole were analyzed.RESULTS & CONCLUSIONS: 5 dosage forms of albendazole are available in the market: tablets,capsules,granules,oral liquid,and syrup.The sales amount of albendazole in recent years dominated the first 3 places in domestic antiparasites market.There are a total of 39 domestic manufactures for albendazole,which can be used to treat 21 kinds of human parasitosis.Its clinical application is widespread.However,severe adverse drug reactions such as the rare encephalitis syndrome and allergic shock etc induced by albendazole pose hidden risks for treatment;therefore,management on its use should be strengthened.

Objective:To investigate the relationship between the pathogeny and expression of CD28 B7 costimulatory molecules on peripheral blood cells in myasthenia gravis.Methods:Expression of CD28?CTLA4 and their ligants B7 1?B7 2 on CD4 + and CD8 + T lymphocytes in 18 patients with MG and 16 healthy controls were examined by flow cytometry.Results:In MG group,the expression of CD28?CTLA4?B7 1?B7 2 molecules were increased.The increased CD28 + cells were mainly of CD4 +T lymphocytes subset.The increased CTLA4 + cells were mainly of CD8 + T lymphocyte subset;There was no significant difference in the expressions of B7.1 and B7.2 molecules on CD4 + or CD8 + T lymphocytes between two groups.Conclusion:The pathogeny of myasthenia gravia is associated with the increased expression of CD28 B7 costimulatory molecules.Examining of CD28 B7 costimulatory molecules on peripheral blood cells may reflect the immune state of MG patients,and is helpful to the diagnosis and treatment.

Objective To investigate the effects of valproic acid ( VPA ) on SD rats with experimental autoimmune encephalomyelitis ( EAE) and its possible immunomodulatory mechanism .Meth-ods Fifty female Sprague-Dawley ( SD) rats were randomly divided into five groups by random digit table , including control group (n=10), EAE group(n=10), low dose VPA treated group (100 mg/kg, n=10), median dose VPA treated group (300 mg/kg, n=10) and high dose VPA treated group (600 mg/kg, n=10).The SD rat model of EAE was induced by immunizing with a guinea pigs′spinal cord homogenate (GPSCH).Normal saline and various doses of VPA were given to rats in according groups twice a day from day 0 to day 19 ( close to the peak stage of EAE ) .The severity of EAE was scored according to the signs and symptoms.Pathological changes were observed through Hematoxylin-Eosin staining, and then the degrees of inflammatory infiltration were evaluated .The numbers of activated neuroglia that expressed Iba-1 in cerebral and lumber cords were counted by immunohistochemistry .The expression of IFN-γ, IL-17 and IL-10 in cer-ebral and lumber cords were measured by ELISA .Results Compared with EAE group , rats in the low, me-dian and high dose VPA treated groups had lower incidence of EAE and prolonged latency , but only the me-dian dose treated group showed significant alleviation in clinical symptoms (P<0.05).Both the median and the high dose treated group showed decreased inflammatory cell infiltration in CNS (P<0.05).Immunohisto-chemistry results showed that the numbers of activated microglia were significantly inhibited in rats treated with median and high dose of VPA in comparison with those in EAE group (P<0.05).Results of ELISA demonstrated that the expression of IFN-γand IL-17 in both median and high dose VPA treated groups were significantly decreased compared with those in EAE group (P<0.05), but only the median dose treated group showed a remarkably increased expression of IL-10 (P<0.05).Conclusion VPA, especially medi-um dose of VPA ( 300 mg/kg ) , had neuroprotective effects on rats with EAE .The possible mechanism might be associated with the inhibited activation of microglia and the increased percentage of anti -inflammato-ry cytokines .

Objective To explore the immunomodulatory mechanism of 2-(-2-benzofuranyl )-2-imidazoline(2-BFI) in rat model of experimental autoimmune encephalomyelitis (EAE).Methods Fifty fe-male Sprague-Dawley(SD) rats were randomly divided into five groups by random digit table , including con-trol group(n=10), EAE model group(n=10), low dose 2-BFI group(1.5 mg/kg, n=10), median dose 2-BFI group (3 mg/kg, n=10) and high dose 2-BFI group (6 mg/kg, n=10).The SD rat model of EAE was induced by immunizing with a guinea pigs′spinal cord homogenate ( GPSCH ) .The severity of EAE was scored according to the signs and symptoms .Pathological changes were observed through Hematoxylin-eosin staining, and then the degrees of inflammatory infiltration were evaluated .The number of activated neuroglia that expressed GFAP and iba 1 in lumbar cords was counted by immunohistochemistry .The expressions of IL-1β, IFN-γ, IL-4 and IL-10 in cervical cords were measured by ELISA .Results Compared with EAE group, rats in the low, median and high dose 2-BFI treatment group had lower incidence of EAE , prolonged latency and decreased CNS inflammation , but only the median dose group showed significant alleviation in clinical symptoms and decrease in CNS inflammatory cell infiltration (P<0.05).Immunohistochemical re-sults showed that the numbers of activated microglia were significantly inhibited ,but the numbers of activated astroglia were increased in the rats treated with median dose of 2-BFI in comparison with the EAE group ( P<0.05).Results of ELISA demonstrated that expressions of IL-1βand IFN-γin 3 mg/kg 2-BFI treated group were significantly decreased compared with that in the EAE group (P<0.05), but expressions of IL-4 and IL-10 were remarkably increased(P<0.05).Conclusion 2-BFI at median dose of 3 mg/kg has a benefi-cial neuroprotective effect on rats with EAE , and its mechanism might be related to immunodulation .

Objective To establish the animal model of experimental allergic encephalomyelitis(EAE)in Lewis rats.Methods The EAE was induced by giving hypodermal injections in feet with spinal cords homogenate of guinea pigs(GPSCH)and complete Freund's adjuvant(CFA)containing 8mg.ml-1 bacille calmette guerin(BCG).The EAE model was evaluated by clinical manifestations and pathologic findings which was studied with the aid of light and electron microscope.Results The EAE in Lewis rats had an incidence of 9/10,a latency of 12.6?1.01d.Bordetella pertussis vaccine(BPV)could increase the morbidity,shorten the latency(P

Objective:To explore effects of Idazoxan(IDA) on changes of neuroglial cells in spinal cords of rats with experimental autoimmune encephalomyelitis(EAE).Methods:Rat EAE was induced by immunization with spinal cord homogenates of Geania pigs.EAE clinical manifestations were assessed in terms of the scoring standards.Histological investigation and immunohistochemistry were observed for the inflammatory dedmylinative lesion of CNS and morphology of glial cells.Results:Ida could not decrease the incidence of EAE,but alleviate its clinical manifestation and histological changes.On day 15 after immunization,astrocytes in and around the inflammatory dedmylinative lesion of CNS in EAE rats treated with Ida increased in number and size,on the contrary,microglia decreased in number and size.Conclusion:Ida has protective effects on EAE and its functional mechanism may be concerned with modulation of immunological mechanism of CNS.

Objective:To review literature reports of varieties of ADRs induced by mebendazole to provide a sci- entific foundation for clinical revaluation of mebendazole.Method:The related literatures in the internal and external medi- cine medical database in 1994-2004 were explored,and then both analysis and statistics were conducted with the methods of epidemiolngy and literature analysis.Result:ADRs induced by mebendazole could be involved in multiple organs.Most victims were children and the elderly.Their latent periods were determined by the types of ADRs,which had a variety of forms.Conclusion:Mebendazole was potentially unsafe.So we should strengthen our rational drug use and post-marketing revaluation of safety.

Objective To investigate the relationship between blood pressure(BP) and prognosis in three different ischemic stroke subtypes.Methods The consecutive patients with a brain infarction proven on diffusion-weighted MRI who were hospitalized within 48 hours after stroke onset between April 2007 and April 2008 were registered.All subjects with acute ischemic stroke consecutively admitted to the neurological wards of the First Affiliated Hospital of Wenzhou Medical College,were registered in the Wenzhou Stroke Registry Program.Data were collected and coded at primary registration.The BP levels were studied during the initial 7 hospital days.Survival and dependency were assessed at 3 months.Outcomes were adjusted age,consciousness level,admission NIHSS score,the decline level of systolic BP,the decline level of diastolic BP,complication and so on. Logistic regression model was used to estimate the relationship between BP and prognosis.Results A U-shaped effect was observed in each subgroup between BP and prognosis.In the subgroups of atherothrombosis,cardioembolism and small artery disease,those who had a BP of 150/95 mm Hg(1 mm Hg=0.133 kPa)on admission,140/90 mm Hg on day 1-7 would have a better prognosis.In the subgroups of atherothrombosis and cardioembolism,the decrease of BP during the first 24 hours was the independent predictor of the death and disability at 3-month.In the atherothrombosis group,when the decrease of systolic BP during the first 24 hours was greater than 20 mm Hg,the risk of the death and disability at 3-month increased 4.44 times(OR 4.44,95%CI 1.70-11.59,P=0.002).In the atherothrombosis group,when the decrease of diastolic BP during the first 24 hours was greater than 10 mm Hg,the risk of the death/disability at 3-month increased 3.70 times(OR 3.70,95%CI 1.54-8.90.P=0.00).In the cardioembolism group,the risk increased respectively 7.98 times(OR 7.98,95%CI 1.34-47.66.P=0.026)and 6.68 times(OR 6.68.95%CI 1.55-28.79,P=0.01).In the subgroups of small artery disease,the decrease of BP during the first 24 hours was not the independent predictor of the death and disability at 3-month.Conclusions A U-shaped effect is observed in each subgroup between BP and prognosis.In the subgroups of atherothrombosis and cardioembolism,the decrease of BP during the first 24 hours is the independent predictor of the death and disability at 3-month.

[ ABSTRACT ] AIM: To study the effect of idazoxan on the permeability of inflammatory blood-brain barrier ( BBB) model in vitro and the expression of tight junction protein ZO-1.METHODS:In vitro BBB model was established by murine brain endothelial cell line bEnd.3 incubated for 7 d.The cells were treated with TNF-α(10 nmol/L) for addi-tional 24 h to establish the inflammatory BBB model, which was pretreated with IDA at doses of 50, 100 and 200μmol/L, respectively.The permeability was measured using fluorescein isothiocyanate-conjugated dextran (FD-40, MW 40,000), the expression of ZO-1 was detected by Western blot analysis, the distribution of ZO-1 was observed by immunofluores-cence, and the mRNA expression of MMP-9/TIMP-1 was measured by RT-PCR.RESULTS:After incubated for 7 d, b. End3 cells converged to be confluent monolayer with low permeability.The inflammatory BBB model induced by TNF-αtreatment displayed much higher permeability with decreased expression of tight junction protein ZO-1, destroyed distribu-tion of ZO-1 and increased mRNA expression of MMP-9.When pretreated with IDA, the permeability was greatly de-creased, the expression of ZO-1 was greatly increased, the abnormal distribution of ZO-1 was greatly ameliorated and the mRNA expression of MMP-9 was obviously reduced.The effect was most significant in IDA ( 200 μmol/L )-pretreated group (P<0.01).CONCLUSION:IDA directly acts on brain endothelial cells to reduce the expression of MMP-9, in-crease the expression of tight junction protein ZO-1 and ameliorate the destroyed distribution of ZO-1 in the inflammatory BBB, thus reversing the abnormally elevated permeability in a inflammatory BBB model in vitro induced by TNF-α.