脑缺血再灌流大鼠脑胶质源性神经营养因子基因表达的变化

探讨脑缺血再灌流不同时程及不同程度缺血对海马及皮层胶质源性神经营养因子（ｇｌｉａｌｃｅｌｌｌｉｎｅ ｄｅｒｉｖｅｄ ｎｅｕｒｏｔｒｏｐｈｉｃ ｆａｃｔｏｒ, ＧＤＮＦ）基因表达的影响,以及Ｎ甲基Ｄ天冬氨酸（Ｎｍ ｅｔｈｙｌＤｓａｐａｒｔａｔｅ, ＮＭＤＡ）受体拮抗剂,钙离子通道阻断剂是否能调节缺血病态下ＧＤＮＦｍ ＲＮＡ的表达。参照Ｓｍ ｉｔｈ 等方法建立大鼠前脑缺血再灌流动物模型。用ＤＩＧＯｌｉｇｏｎｕｃｌｅｏｔｉｄｅ ３′ｅｎｄ ｌａｂｅｌｉｎｇ Ｋｉｔ,标记５１ ｍ ｅｒ的ＧＤＮＦ寡核苷酸探针在含有海马结构的冰冻组织切片上进行原位杂交检测ＧＤＮＦｍ ＲＮＡ的表达。１０ ｍ ｉｎ 缺血再灌流２ ｈ,齿状回ＧＤＮＦｍ ＲＮＡ表达上调。再灌流６ ｈ,ＣＡ１,ＣＡ３ 和皮层ＰＡＲ区ＧＤＮＦｍ ＲＮＡ表达亦见增多,２４ ｈ 达高峰。Ｋｅｔａｍ ｉｎｅ 可使ＧＤＮＦ的基因表达在海马结构及皮层ＰＡＲ区明显低于相应的缺血再灌流组,统计学差异显著（Ｐ＜ ００５）。脑缺血再灌流时ＧＤＮＦ基因表达增加,对缺血神经元可能起保护作用。Ｋｅｔａｍ ｉｎｅ可阻断缺血后ＧＤＮＦｍ ＲＮＡ 的表达增加,提示ＮＭＤＡ谷氨酸受体很可能参与介导了缺     

癫痫状态大鼠海马GAD67mRNA,GABA—TmRNA表达水平的动态观察

为了研究癫痫状态下大鼠海马区ＧＡＤ６７ｍＲＮＡ、ＧＡＢＡ－ＴｍＲＮＡ的表达水平，采用马桑内酯（ｃｏｒｉａｒｉａｌａｃｔｏｎｅ，ＣＬ）侧脑室注射所致大鼠癫痫状态模型，动态观察ＣＬ致痫前后大鼠海马ＧＡＤ６７ｍＲＮＡ、ＧＡＢＡ－ＴｍＲＮＡ表达水平的变化，同时观察了动物行为学改变及脑电图变化。实验结果显示，侧脑室注射ＣＬ后约１５分钟，大鼠海马及皮质ＥＥＧ出现阵发性棘慢波及高波幅慢波；ＣＬ注射后１５～３０分钟，实验鼠均出现明显的连续发作性四肢抽动、尖叫等症状，且持续６～８小时。ＣＬ侧脑室注射后３０分钟、１小时、２小时、４小时或８小时各时限点组大鼠海马ＧＡＤ６７ｍＲＮＡ表达水平均低于正常对照组。ＣＬ注射后３０分钟组大鼠海马ＧＡＢＡ－ＴｍＲＮＡ表达水平高于正常对照组，但３０分钟后各时限组ＧＡＢＡ－ＴｍＲＮＡ表达水平与正常对照组比较增加不明显。提示ＣＬ所致癫痫持续状态的后期，大鼠海马ＧＡＤ６７ｍＲＮＡ、ＧＡＢＡ－ＴｍＲＮＡ表达信号的减弱，可能与致痫因素及癫痫持续后期合并的缺血、缺氧性脑损害致使海马区ＧＡＤ、ＧＨＢＡ－Ｔ免疫反应阳性细胞死亡有关。     

氟美松对成年大鼠持续性局灶脑缺血后细胞凋亡和Fas基因表达的作用

目的 研究氟美松对成年人局灶性脑缺血后细胞凋亡及相关Ｆａｓ基因表达的作用。方法 健康雄性ＳＤ大鼠随机分成１４组（每组５只）。１～７组为对照组，８～１４组为实验组。用右侧近端大脑中动脉电凝术建立大鼠持续性局灶性脑缺血模型。缺血后１ｈ，实验组动物尾静脉射氟美松（５ｍｇ／ｋｇ），对照组注射生理盐水。分别在缺血后３、６、１２、２４、４８、７２和１２０ｈ取材，用原位末端标记（ＴＵＮＥＬ法）、原位ＲＴ－ＰＣＲ法分别检测缺血后细胞凋亡和Ｆａｓ ｍＲＮＡ表达并进行了半定量分析。结果 氟美松可导致缺血后细胞凋亡提前发生，促进大鼠局灶性脑缺血后Ｆａｓ ｍＲＮＡ的表达，使其表达开始时间提前、表达持续时间延长、表达细胞增多及表达信号增强。结论 氟美松可促缺血后细胞凋亡相关基因Ｆａｓ ｍＲＮＡ的表达，可能是其加重成年大鼠缺血性脑损伤尤其     

急性脑梗死患者白细胞变形能力、粘附功能和粘附分子表达的变化

目的探讨白细胞变形能力、粘附功能和白细胞粘附分子与急性脑梗死发生的关系。方法检测了６８例急性脑梗死患者急性期（发病３、７、１５天）和恢复期（２１、３０天）、４２例脑出血患者和６０名健康人外周血白细胞滤过指数（ＬＦＩ）、白细胞粘附率（ＬＡＲ）、白细胞ＣＤ１８表达和血清可溶性细胞间粘附分子１（ｓＩＣＡＭ１）浓度变化。结果急性脑梗死患者ＬＦＩ、ＬＡＲ、ＣＤ１８表达和ｓＩＣＡＭ１浓度均明显增高，与对照组和脑出血组比较差异有显著意义；脑梗死患者恢复期ＬＦＩ、ＬＡＲ、ＣＤ１８表达和ｓＩＣＡＭ１浓度明显低于急性期。随着病情好转，ＬＦＩ、ＬＡＲ、ＣＤ１８表达和ｓＩＣＡＭ１浓度逐渐降低，第３、７、１５、２１、３０天各指标比较差异有极显著意义。有并发症者各指标变化较无并发症者更明显。急性脑梗死患者ＬＦＩ、ＬＡＲ与ＣＤ１８表达和ｓＩＣＡＭ１浓度呈正相关。ＬＦＩ与ＬＡＲ呈正相关。结论急性脑梗死患者白细胞变形能力降低、粘附功能和ＣＤ１８表达及可溶性细胞间粘附分子１浓度增高，可能与急性脑梗死的发生、发展有关     

神经生长因子对大鼠脑缺血后海马CA1区神经细胞凋亡的影响

目的　研究神经生长因子（ Ｎ Ｇ Ｆ）在缺血性脑损伤中对神经细胞凋亡的影响。方法　采用大鼠 ４ 血管闭塞脑缺血模型， Ｔ Ｕ Ｎ Ｅ Ｌ 方法原位标记 Ｄ Ｎ Ａ 片段，观察脑室内注射 Ｎ Ｇ Ｆ 后海马 Ｃ Ａ１ 区神经细胞凋亡的变化。结果　使用 Ｎ Ｇ Ｆ 后 ３ｄ 海马 Ｃ Ａ１ 区 Ｔ Ｕ Ｎ Ｅ Ｌ 阳性神经细胞数为神经细胞总数的１３．９％ ±１１．６％ ，与缺血对照组（２１．３％ ±１３．７％ ）比较显著减少（ Ｐ＜ ０．０１）。结论　外源性 Ｎ Ｇ Ｆ 对脑缺血所致的神经细胞凋亡可能具有一定的保护作用。     

重症肌无力患者外周血乙酰胆碱受体特异性T细胞sIL-2R、IFN-γ、IL-4的表达和转录

目的了解ＭＧ患者的乙酰胆碱受体（ＡＣｈＲ）特异性细胞免疫应答。方法采用酶联免疫吸附试验（ＥＬＩＳＡ）检测３０例ＭＧ患者和２０名健康对照者经ＡＣｈＲ刺激后外周血单核细胞（ＰＢＭＣ），辅助性Ｔ细胞１（Ｔｈ１）相关的干扰素（ＩＦＮ）γ、辅助性Ｔ细胞２（Ｔｈ２）相关的白细胞介素（ＩＬ）４及与细胞免疫活化密切相关的可溶性白细胞介素２受体（ｓＩＬ２Ｒ）的分泌，用逆转录聚合酶链反应（ＲＴＰＣＲ）结合狭缝印迹杂交检测ＩＦＮγ、ＩＬ４的信息核糖核酸（ｍＲＮＡ）转录。结果总ＭＧ患者组ＩＦＮγ显著高于健康对照组，尤以急性ＭＧ患者组（１４例）更明显，且其升高与其ｍＲＮＡ转录水平一致。虽然这两组ＭＧ患者的ｓＩＬ２Ｒ亦明显高于健康对照组，但患者组的ＩＬ４表达及其ｍＲＮＡ转录与健康对照组相比差异无显著意义。结论ＭＧ患者有Ｔｈ１和Ｔｈ２的失衡，ＭＧ患者有ＡＣｈＲ特异性细胞免疫活化，Ｔｈ１的细胞因子ＩＦＮγ可能作为效应因子参与了ＭＧ的发病。     

大鼠脑缺血再灌流后HSP70的表达和bFGF对其影响

目的研究脑缺血再灌流后热休克蛋白７０（ＨＳＰ７０）表达的意义及外源性碱性成纤维细胞生长因子（ｂＦＧＦ）对其影响。方法应用免疫组化的方法观察大鼠局灶性脑缺血再灌流后脑组织ＨＳＰ７０的表达以及在侧脑室注射外源性ｂＦＧＦ对其影响。结果缺血２小时再灌流０小时即可见ＨＳＰ７０表达,至２４小时达高峰。ｂＦＧＦ组与生理盐水组相比于６～４８小时各组可见ＨＳＰ７０表达增高（Ｐ〈０．０５）。结论脑缺血诱导ＨＳＰ７０的     

许旺细胞的IGF-Ⅰ基因转染及其mRNA和蛋白表达

为探索类胰岛素生长因子Ⅰ（ＩＧＦⅠ）基因转染许旺细胞的可能性,采用体外培养许旺细胞（ＳＣｓ）,构建ＩＧＦⅠ逆转录病毒载体,利用基因转染技术将ＩＧＦⅠ基因转入ＳＣｓ,核酸分子杂交及细胞免疫组化检测ＳＣｓ 表达ＩＧＦⅠｍ ＲＮＡ及其蛋白水平,辅以正常ＳＣｓ作对照。结果：ｐＬＸＳＮＩＧＦⅠ转染的ＳＣｓ在体外相同培养条件下较正常ＳＣｓ 表达ＩＧＦⅠｍ ＲＮＡ 及其蛋白水平能力分别增强４８ 和３２ 倍,均有显著意义。研究表明,利用基因转染技术可将外源性ＩＧＦⅠ基因转入ＳＣｓ,提高ＳＣｓ中的ＩＧＦⅠ的含量和活性     

兴奋性氨基酸引起星形细胞肿胀的机理探讨

用［３Ｈ］－３－氧－甲基－Ｄ－葡萄糖摄取法测定细胞体积，观察不同类型的谷氨酸受体激动剂对体外培养星形细胞水含量的影响，并观察影响肿胀的有关因素以及中药丹参有效成分７６４－３对细胞肿胀的保护作用。结果表明谷氨酸引起的星形细胞肿胀是剂量依赖性和钙依赖性的，与细胞外液是否有糖无关，培养基中加ｄＢｃＡＭＰ使谷氨酸的作用更明显；代谢型谷氨酸受体激动剂ｔｒａｎｓ－ＡＣＰＤｌｍＭ可以引起细胞肿胀，而离子型谷氨酸受体激动剂ＡＭＰＡｍｍｏｌ不引起肿胀，Ｌ－ＡＰ３（代谢型谷氨酸受体拮抗剂）和７６４－３可以对抗ｔｒａｎｓ－ＡＣＰＤ引起的肿胀。结果提示细胞内信使系统参与星形细胞的体积调节。     

许旺细胞的IGF—I基因转染及其mRNA和蛋白表达

为探索类胰岛素生长因子－Ⅰ（ＩＧＦ－Ⅰ）基因转染旺细胞的可能性，采用体外培养许旺细胞（ＳＣｓ），构建ＩＧＦ－Ⅰ逆转录病毒载体，利用基因转梁技术将ＩＧＦ－Ⅰ基因转入ＳＣｓ，核酸分子杂交及细胞免疫组化检测ＳＣｓ表达ＩＧＦ－Ⅰ ｍＲＮＡ及其蛋白水平，辅以正常ＳＣｓ作对照。结果：ｐＬＸＳＮ－ＩＧＦ－Ⅰ转染的ＳＣｓ在体外相同培养条件下较正常ＳＣｓ表达ＩＧＦⅠ ｍＲＮＡ及其蛋白水平能力分别增强４．８和３．２倍     

视频脑电图在小儿癫痫诊断中的应用

目的评价视频脑电图(video-EEG)在小儿癫诊断中的应用价值。方法对126例具有发作性症状的患儿进行连续8h的包括清醒、睡眠、诱发试验及必要的认知测验的视频脑电图监测。结果经发作期视频脑电图证实,39例初诊为癫性发作的患儿中14例(35%)为非癫性发作;15例其他症状发作中13例(86%)为非癫性发作。64例样放电患儿中51例(80%)确定发作类型,22例(34%)确定癫类型。视频脑电图可发现短暂轻微的癫发作及样放电引起的一过性认知损伤。结论视频脑电图在排除非癫性发作、确定癫性发作的类型、评价脑电-临床关系方面可提供准确可靠的证据,进一步提高癫的临床诊断水平。     

Role of glutathione in repair of free radical damage in hippocampus in vitro.

Depletion of glutathione (GSH), an intrinsic antioxidant, increases vulnerability to free radical damage in a number of cell systems. This study investigates the role of GSH in limiting electrophysiological damage and/or recovery from free radical exposure in slices of guinea pig hippocampus. Synaptic potentials (PSPs) and population spikes (PSs) were recorded from field CA1. Free radicals were generated from 0.006% peroxide through the Fenton reaction. Analysis of the input-output curves showed that peroxide treatment decreased PSPs and impaired ability of the PSPs to generate PSs as previously reported. Recovery was nearly total within a half hour. Treatment with 5 mM buthionine sulfoximine (BSO) for 2 h depleted hippocampal GSH to 79.2% of control values. The extent of free radical damage was not increased. Recovery, however, was only partial. GSH was further depleted by oxidation with diamide or covalent bonding with dimethyl fumarate (DMF) immediately before and during the peroxide treatment. Neither diamide nor DMF treatment in BSO-incubated tissue enhanced peroxide-induced electrophysiological deficits. Following these treatments, however, tissue showed little recovery from free radical damage. We conclude that glutathione is essential for repair processes in hippocampal neurons exposed to oxidative damage.     

Neuroprotective properties of memantine in different in vitro and in vivo models of excitotoxicity

The pathogenesis of stroke, trauma and chronic degenerative diseases, such as Alzheimer's disease (AD), has been linked to excitotoxic processes due to inappropriate stimulation of the N-methyl-D-aspartate receptor (NMDA-R). Attempts to use potent competitive NMDA-R antagonists as neuroprotectants have shown serious side-effects in patients. As an alternative approach, we were interested in the anti-excitotoxic properties of memantine, a well-tolerated low affinity uncompetitive NMDA-R antagonist presently used as an anti-dementia agent. We explored in a series of models of increasing complexity, whether this voltage-dependent channel blocker had neuroprotective properties at clinically relevant concentrations. As expected, memantine protected neurons in organotypic hippocampal slices or dissociated cultures from direct NMDA-induced excitotoxicity. However, low concentrations of memantine were also effective in neuronal (cortical neurons and cerebellar granule cells) stress models dependent on endogenous glutamate stimulation and mitochondrial stress, i.e. exposure to hypoxia, the mitochondrial toxin 1-methyl-4-phenylpyridinium (MPP+) or a nitric oxide (NO) donor. Furthermore, memantine reduced lethality and brain damage in vivo in a model of neonatal hypoxia-ischemia (HI). Finally, we investigated functional rescue (neuronal capacity to migrate along radial glia) by memantine in cerebellar microexplant cultures exposed to the indirect excitotoxin 3-nitropropionic acid (3-NP). Potent NMDA-R antagonists, such as (+)MK-801, are known to block neuronal migration in microexplant cultures. Interestingly, memantine significantly restored the number of neurons able to migrate out of the stressed microexplants. These findings suggest that inhibition of the NMDA-R by memantine is sufficient to block excitotoxicity, while still allowing some degree of signalling.     

Knowledge of management of epilepsy in young adults in Jordan

PURPOSE: Nationwide studies on public knowledge of epilepsy have been undertaken in several countries, but not in Jordan. The purpose of this study was to evaluate knowledge of the management of epilepsy in Jordan. METHODS: A cross-sectional study was performed during the period February-June 2005 on 16,044 individuals selected randomly to represent all regions of Jordan. Respondents were interviewed and asked to complete a five-item questionnaire testing their knowledge of the management of epilepsy. RESULTS: Most of the individuals (77.9%) agreed that patients with epilepsy are best treated in a specialized hospital for neurological diseases. Although the largest percentage of the sample (86.7%) agreed that the best person to treat epilepsy is a neurologist, others believed that a hypnotherapist (25.4%) or a religious healer (22.6%) is. As an immediate measure during an attack of epilepsy, most respondents (82.6%) agreed that protecting the patient's head is most important. During the postseizure period, 45.8% of respondents believed that offering the patient water or a cold or hot drink is appropriate. Two-thirds of the respondents (66.2%) agreed that epilepsy can be treated with drugs. Responses of the participants to the five items significantly differed with respect to age, gender, level of education, and occupation. CONCLUSION: Jordanians are reasonably well informed about most aspects of the management of epilepsy, but there is still a need for public education about this disorder.