电针百会和大椎穴对癫痫大鼠海马CA3和DG区ephrinA5表达的影响

目的探讨电针百会和大椎穴对颞叶癫痫大鼠海马组织中CA3和DG区ephrin A5的调控作用。方法将30只SpragueDawley(SD)大鼠随机分为对照组、癫痫组和电针+癫痫组,每组各10只。建立氯化锂―匹罗卡品颞叶癫痫大鼠模型。造模成功的大鼠电针百会和大椎穴治疗8周后,分别取3组大鼠海马组织,采用实时荧光定量PCR(q RT-PCR)检测各组大鼠海马CA3和DG区ephrin A5 m RNA水平表达变化;采用Western blotting和免疫组织化学(免疫组化)法检测各组大鼠海马CA3和DG区ephrin A5蛋白水平表达变化。结果 q RT-PCR结果显示:与对照组相比,癫痫组大鼠海马组织中ephrin A5 m RNA表达下调(P <0.05)。通过8周电针百会和大椎穴连续治疗后,ephrin A5 m RNA水平上调(P <0.05)。Western blotting结果显示:ephrin A5蛋白水平变化趋势与m RNA水平相一致。免疫组化结果显示:在CA3区,癫痫组ephrin A5蛋白水平下调;电针后ephrin A5蛋白水平上调。而在DG区与对照组相比,癫痫组和电针+癫痫组,ephrin A5蛋白水平变化不明显。结论电针百会和大椎穴的抗癫痫作用机制很可能与ephrin A5在海马CA3区中的调控机制密切相关。     

低频重复经颅磁刺激预处理对毛果芸香碱致痫大鼠海马GAD65、NMDAR1表达的影响

目的探讨低频重复经颅磁刺激(rTMS)对毛果芸香碱致痫作用的影响及其机制。方法将大鼠随机分为rTMS刺激组、假刺激组和对照组,分别进行连续2周的相应预处理后,制作氯化锂-毛果芸香碱急性癫痫(EP)模型,观察造模后90min内痫性发作潜伏期和痫性发作行为表现。应用免疫组化方法观察各组大鼠海马区谷氨酸脱羧酶65(GAD65)、N-甲基-D-天门冬氨酸受体1(NMDAR1)的表达。结果rTMS刺激组大鼠痫性发作潜伏期较假刺激组和对照组延长,发作程度减轻(均P<0.05);rTMS刺激组大鼠海马区神经元GAD65的表达量明显高于对照组,NMDAR1表达量明显低于对照组(均P<0.05),但假刺激组与对照组二者表达差异无统计学意义(P>0.05)。结论低频rTMS预处理有明显抗痫作用,可能通过调节海马区GAD65和NMDAR1表达而起作用。     

GAD在颞叶癫痫大鼠海马内源性促痫机制中的作用

目的:探讨GAD65、GAD67在颞叶癫痫发生后海马内源性促痫机制中的作用。方法:112只雄性SD大鼠随机分为实验组(n=70)与对照组(n=42),实验组大鼠选用海人酸腹腔注射法建立颞叶癫痫模型,对照组大鼠腹腔注射无菌生理盐水。选取腹腔注射后3小时、6小时、12小时、24小时、48小时、7天、30天为研究的时间点,颞叶海马的CA1区、CA3区、齿状回为研究部位。腹腔给药后每天观察大鼠的行为学变化,大鼠处死前进行EEG描记。用原位杂交方法检测不同时间点海马不同区域GAD65、GAD67mRNA的表达,免疫组织化学法检测GAD65、GAD67蛋白的表达。结果:实验组大鼠海马GAD65 mRNA及其蛋白的表达随时间呈逐渐增高趋势,致痫后48小时～30天,GAD65 mRNA及其蛋白表达较对照组增高(48小时P<0.05;7～30天P<0.01);海人酸致痫后6小时、24小时实验组大鼠海马的GAD67mRNA及其蛋白表达较对照组增高(分别为P<0.01、P<0.05)。结论:颞叶癫痫急性期海马GAD67表达的增高及慢性期海马GAD65表达的增高是癫痫发生后机体的内源性抗痫机制。     

GAD67/GAD65在颞叶癫痫大鼠海马内源性促痫机制中的作用

目的探讨GAD67/GAD65在颞叶癫痫发生后大鼠海马内源性促痫机制中的作用.方法112只雄性SD大鼠随机分为实验组(n=70)与对照组(n=42),实验组大鼠选用海人酸腹腔注射法建立颞叶癫痫模型,对照组大鼠腹腔注射无菌生理盐水.选取腹腔注射后3 h、6 h、12 h、24 h、48 h、7 d、30 d为研究的时间点,颞叶海马的CA1区、CA3区、齿状回为研究部位.腹腔给药后每天观察大鼠的行为学变化,大鼠处死前进行EEG描记.免疫组织化学法检测GAD65、GAD67蛋白的表达.结果海人酸致痫后6 h,实验组大鼠海马CA1区、CA3区GAD67/GAD65的比率较对照组升高(P＜0.01);海人酸致痫后30 d,实验组大鼠海马齿状回GAD67/GAD65的比率较对照组降低(P＜0.05).结论颞叶癫痫急性期CA1区、CA3区GAD67/GAD65比率的增高及慢性期齿状回GAD67/GAD65比率的降低与颞叶癫痫发生及癫痫发生后机体的内源性抗痫机制密切相关.     

γ-氨基丁酸能神经元在新生大鼠缺氧性痫性发作中的作用

目的 通过观察新生大鼠早期发育过程中及缺氧性痫性发作后海马组织病理改变以及原癌基因c-fos蛋白、谷氨酸脱羧酶(glutamate decarboxylase,GAD)的变化,探讨γ-氨基丁酸(γ-amino butylic acid,GABA)能神经元在缺氧性痫性发作中的作用及可能的影响机制.方法 采用出生后10d的SD大鼠建立改良Jensen缺氧诱导痫性发作模型,分为痫性发作后1d、3d、7d、14d 4组,并选取相应时间点正常大鼠为对照组,采用尼氏染色方法检测海马组织的组织病理变化,免疫组织化学法检测各组海马c-fos蛋白灰度值以及GAD阳性神经元数量的改变.结果 尼氏染色结果显示,各缺氧性痫性发作组海马区形态结构正常,细胞排列略稀疏,但未见明显的细胞丢失.免疫组化结果显示,与对照组比较,c-fos蛋白灰度值在痫性发作后7d,在缺氧性痫性发作组海马CA2、CA3和DG区明显地降低(P ＜0.05);GAD阳性细胞数在痫性发作后7～ 14d,缺氧性痫性发作组海马CA1、CA3和DG区明显地减少(P＜0.05).结论 缺氧性痫性发作后14d内并没有造成大鼠海马区及时或迟发性细胞丢失,但c-fos表达在大鼠海马区有迟发性增高;缺氧性痫性发作后海马GABA神经元数量的减少可能是新生大鼠缺氧诱导痫性发作后癫痫易感性升高的原因之一.     

癫痫状态大鼠海马HSP70 mRNA及GAD67 mRNA表达水平的研究

目的研究癫痫演变过程中特定脑区选择性神经元损伤与修复机制。方法采用分子杂交方法动态观察了马桑内酯(coriariaLactone,CL)诱导的癫痫状态大鼠海马区早期神经元损伤标志物热休克蛋白(heatshockprotein,HSP)70mRNA及谷氨酸脱羧酶(glutaminaciddecarboxylase,GAD)67mRNA表达水平的变化,同时也观察了大鼠行为学及脑电图改变。结果侧脑室注射CL后20分钟,大鼠出现连续发作的四肢抽搐伴有EEG持续性的棘慢波及尖波发放。注射CL后2小时组大鼠海马GAD67mRNA表达水平有所降低,但HSP70mRNA表达变化不明显,注射CL后8小时组大鼠海马GAD67mRNA表达水平明显低于对照组(P＜0.01),HSP70mRNA表达水平较对照组明显增高(P＜0.01)。结论癫痫状态期间大鼠海马区可能存在神经元损伤与修复过程,而且GABA能神经元受到不同程度的损伤。     

癫痫状态大鼠海马HSP70mRNA及GAD67mRNA表达水平的研究

目的 研究癫痫演变过程中特定脑区选择性神经元损伤与修复机制。方法 采用分子杂交方法动态观察了马桑内酯（coriaria Lactone,CL)诱导的癫痫状态大鼠海马区早期神经元损伤标志物热休克蛋白（heat shock protein,HSP)70mRNA及谷氨酸脱羧酶（glutamin acid decarboxylase,GAD)67mRNA表达水平的变化,同时也观察了大鼠行为学及脑电图改变。结果 侧脑室注射CL后20分钟,大鼠出现连续发作的四肢抽搐伴有EEG持续性的棘慢波及尖波发放。注射CL后2小时组大鼠海马GAD67 mRNA表达水平有所降低,但HSP70mRNA变化不明显,注射CL后8小时组大鼠海马GAD67mRNA表达水平有所降低,但HSP70mRNA表达变化不明显,注射CL后8小时组大鼠海马GAD67mRNA表达水平明显低于对照组（P＜0．01）,HSP70mRNA表达水平较对照组明显增高（P＜0．01）。结论 癫痫状态期间大鼠海马区可能存在神经元损伤与修复过程,而且GABA能神经元受到不同程度的损伤。     

锂-匹罗卡品诱导癫痫持续状态大鼠海马CA1，CA3和PG区P糖蛋白的动态表达

【背景】药物抗性癫痫的病理生理机制目前还不清楚。现有的证据提示P糖蛋白可能参与了药物抗性癫痫的形成。【目的】观察锂-匹罗卡品诱导的大鼠慢性颞叶内侧癫痫模型中海马不同分区P糖蛋白是否出现过度表达,并进而探讨其表达与癫痫发作频度是否相关。【方法】选择6-8周雌性SD大鼠,予锂-匹罗卡品诱导大鼠形成颞叶内侧癫痫慢性模型,对大鼠进行行为学观察及视频脑电记录;Western-Blot、实时定量RT-PCR及免疫组化方法分别检测P糖蛋白在处理组、假处理组、空白对照组中不同时间点（1d及60d）海马不同分区（CA1、CA3及DG）的表达情况。【结果】87.5％（35/40）的大鼠在锂-匹罗卡品诱导的急性期出现惊厥持续状态,伴有与临床癫痫全身发作类似的脑电变化;慢性期出现自主发作,发作间期脑电记录可见到痫性放电;与对照组相比,模型鼠急性期及慢性期在海马CA1、CA3及DG区均出现P糖蛋白的过度表达,增加约70％以上(p<0.05);应用免疫组化染色发现P糖蛋白阳性显色定位于锥体细胞层神经元上。【结论】在慢性颞叶内侧癫痫模型中急性期及慢性期海马锥体细胞层神经元均出现P糖蛋白的过度表达,并证实P糖蛋白的过度表达可能与痫性发作密切相关,但非发现其表达程度与发作频度相关。     

中枢组胺对戊四氮点燃大鼠海马GABA能神经元的影响

目的:探讨中枢组胺抗癫痫的作用机制。方法:应用免疫组织化学方法研究中枢组胺对戊四氮(PTZ)致癫痫大鼠海马神经元GABA、GAD67表达的影响。结果:戊四氮致痫组大鼠海马神经元GABA、GAD67的表达量明显低于正常对照组,L-组胺酸干预组明显高于戊四氮致痫组,L-组胺酸干预组与正常对照组之间差异无显著性意义。结论:中枢组胺通过激活海马GABA能神经元来抑制癫痫的发生和发展。     

雌激素和姜黄素对海人酸杏仁核点燃大鼠行为学、脑电图及海马神经元损伤的影响

目的了解雌激素和姜黄素对海人酸(kainic acid,KA)杏仁核点燃大鼠癫痫发作的影响。方法给去势的雌性大鼠添加雌激素治疗,添加姜黄素治疗,或添加雌激素和姜黄素治疗,比较各组大鼠致痫后癫痫发作的行为学、脑电图和海马神经元损伤的变化。结果给雌激素治疗的大鼠重型发作(Racine 4/5级)评分最高,而雌激素加姜黄素治疗组评分最低(P<0.05)。脑电图的变化与行为学的改变基本一致。致痫后大鼠注射KA侧海马CA3区、CA4区可见到明显的细胞损伤,而该侧海马CA1区、齿状回区(DG)及对侧海马CA3区、CA1区及DG区神经元损害不明显。雌激素组大鼠双侧海马CA3区均出现加重的神经元损害,姜黄素组及雌激素加姜黄素组大鼠海马注射对侧CA3区存活神经元较雌激素组明显增加(P<0.01)。结论高水平的雌激素可以加重癫痫的发作,给姜黄素治疗可以减轻大鼠海马CA3区神经元损害。     

Cellular origin and regulation of D- and L-serine in in vitro and in vivo models of cerebral ischemia

D-Serine is known to be essential for the activation of the N-methyl-D-aspartate (NMDA) receptor in the excitation of glutamatergic neurons, which have critical roles in long-term potentiation and memory formation. D-Serine is also thought to be involved in NMDA receptor-mediated neurotoxicity. The deletion of serine racemase (SRR), which synthesizes D-serine from L-serine, was recently reported to improve ischemic damage in mouse middle cerebral artery occlusion model. However, the cell type in which this phenomenon originates and the regulatory mechanism for D-/L-serine remain elusive. The D-/L-serine content in ischemic brain increased until 20 hours after recanalization and then leveled off gradually. The results of in vitro experiments using cultured cells suggested that D-serine is derived from neurons, while L-serine seems to be released from astroglia. Immunohistochemistry studies of brain tissue after cerebral ischemia showed that SRR is expressed in neurons, and 3-phosphoglycerate dehydrogenase (3-PGDH), which synthesizes L-serine from 3-phosphoglycerate, is located in astrocytes, supporting the results of the in vitro experiments. A western blot analysis showed that neither SRR nor 3-PGDH was upregulated after cerebral ischemia. Therefore, the increase in D-/L-serine was not related to an increase in SRR or 3-PGDH, but to an increase in the substrates of SRR and 3-PGDH.     

新疆维吾尔族胶质瘤中ATM及PTEN蛋白的表达与临床意义

目的探讨新疆维吾尔族(维族)胶质瘤中ATM及PTEN蛋白的表达与临床意义。方法运用免疫组化法检测50例新疆维族胶质瘤中ATM及PTEN蛋白的表达,同时以20例胶质瘤旁组织作为对照,分析其表达的差异及其表达与胶质瘤临床病理特征、患者生存期之间的关系。结果 (1)新疆维族胶质瘤和胶质瘤旁组织中ATM蛋白表达的阳性率分别为60%(30/50)和10%(2/20);PTEN蛋白表达的阳性率分别为30%(15/50)和75%(15/20);(2)ATM及PTEN蛋白表达在胶质瘤和胶质瘤旁组织中的表达差异有统计学意义(均P<0.001);(3)ATM蛋白及PTEN蛋白在WHO分级、不同生存期各组中表达的差异均有统计学意义(均P<0.05),在临床表现、性别、肿瘤大小、肿瘤部位、是否复发各组之间的差异均无统计学意义(均P>0.05)。结论 (1)ATM及PTEN蛋白在新疆维族胶质瘤中及胶质瘤旁组织中表达有差异,表明ATM及PTEN蛋白在胶质瘤发生发展中可能起到一定作用;(2)新疆维族胶质瘤中ATM及PTEN蛋白均与WHO分级和不同生存期表达相关,提示ATM及PTEN蛋白可作为判定胶质瘤生物学行为和预后判断的参考指标。     

Uptake and metabolism of malate in neurons and astrocytes in primary cultures

Uptake and oxidative metabolism of [¹⁴C]malate as well as its incorporation into aspartate, glutamate, glutamine, and GABA were studied in cultured cerebral cortical neurons (GABAergic), cerebellar granule neurons (glutamatergic), and cerebral cortical astrocytes. All cell types exhibited high affinity uptake of malate (K_m 10–85 μM) with slightly higher V_max values in neurons (0.1–0.2 nmol × min^?1 × mg^?1) than in astrocytes (0.06 nmol × min^?1 × mg^?1). Malate was oxidatively metabolized in all three cell types with nominal rates of ¹⁴CO₂ production of 2–15 pmol × min^?1 × mg^?1. The oxidation of malate was only slightly inhibited by 5 mM aminooxyacetic acid (AOAA). In granule cell preparations [¹⁴C]malate was incorporated into aspartate and glutamate and, to a much less extent, into glutamine. This incorporation was blocked by 5 mM AOAA. Astrocytes exhibited slightly higher incorporation rates into aspartate and glutamate, but in these cells glutamine was labelled to a considerable extent. AOAA (5 mM) inhibited the incorporation by 60–70%. In cultures of cerebral cortical neurons, very low levels of radioactivity derived from [¹⁴C]malate were found in aspartate and glutamate, and GABA was not labelled at all. Glutamine had the same specific activity as glutamate, indicating that the low rates of incorporation of radioactivity into amino acids in this preparation is likely to exclusively represent metabolism of malate in the small population of astrocytes (5% of total cell number), contaminating the neuronal cultures. The findings suggest that exogenous malate to a quantitatively limited extent may serve as a precursor for transmitter glutamate in glutamaterigc neurons. Astrocytes are able to metabolize malate to glutamate and related amino acids, but oxidize little malate to CO₂. © Wiley-Liss, Inc.     

重庆市儿童青少年焦虑抑郁发生情况调查

目的 调查重庆市儿童青少年焦虑抑郁发生情况,为儿童青少年学生心理疏导提供参考.方法 通过整群抽样选取儿童青少年学生425名,年龄范围为7～16岁,采用儿童焦虑性情绪障碍筛查量表及儿童抑郁障碍自评量表对被试进行评估.结果 ①34.6％(147/425)的青少年存在焦虑情绪,9.9％(42/425)的青少年存在抑郁情绪;5.6％(24/425)同时存在焦虑抑郁情绪,33.2％(141/425)仅有焦虑或抑郁情绪,61.2％(260/425)无情绪问题.②男性、女性间焦虑抑郁情绪分布差异具有统计学意义(x 2=12.592,P＜0.05),男性中66.1％的无任何情绪问题,25.3％仅存在焦虑情绪,5.6％仅存在抑郁情绪,3.0％存在焦虑抑郁情绪;女性中55.2％的无任何情绪问题,33.3％仅存在焦虑情绪,2.6％仅存在抑郁情绪,8.9％存在焦虑抑郁情绪.不同性别间焦虑得分差异具有统计学意义(t=4.638,P＜0.05),抑郁情绪得分差异无统计学意义(t=0.672,P＞0.05).③年龄与焦虑(r=-0.42,P＞0.05)、抑郁情绪(r=0.071,P＞0.05)间相关关系无统计学意义,焦虑情绪与抑郁情绪评分之间存在相关关系(r=0.420,P＜0.001).结论 儿童青少年焦虑抑郁发生率较高,需要给予积极心理关注.     

Interactions of Phenytoin and Phenobarbital in Terms of Order and Temporal Spacing of Administration in Monkeys

The present study investigated order and temporal spacing interactions of phenytoin and phenobarbital in terms of plasma levels during multiple dosing in monkeys. Phenytoin at a dose of 30 mg/kg and phenobarbital at a dose of 3 mg/kg were administered separately to 4 animals (control group) by nasogastric intubution daily for 10 days. In four subsequent 10-day periods the drugs were administered together in 4 other animals (interaction group) at different times of the day (immediately following one another, 1/2 hr apart, and 6 hr apart) and in a different order of administration (either phenobarbital first and phenytoin later, or the reverse). Blood samples were obtained on the 5th, 8th, and 10th day of each 10-day period. The plasma data indicated: (a) phenytoin is capable of autoinduction, (b) phenobarbital lowers the levels of phenytoin under the four methods of administration studied here, and (c) phenytoin can affect the levels of phenobarbital. The latter interaction is a function of order and temporal spacing of drug administration.     

脱细胞血管基质制备及体内外生物相容性

背景：利用脱细胞血管基质作为血管支架具有以下优点：脱细胞血管基质保留了自然血管的复杂三维结构;脱细胞基质表面的生长因子和结构域有利于细胞的黏附和浸润。 目的：制备脱细胞血管基质并对其体内外生物相容性进行评价。 方法：采用胰蛋白酶、Triton X-100逐步处理猪颈动脉制备脱细胞血管基质。采用皮下植入实验、急性毒性实验和体外细胞毒性实验等评价其生物相容性。 结果与结论：脱细胞基质材料具有良好的化学稳定性,未释放对红细胞产生破坏溶解作用的有害元素,未引起急性溶血反应,对细胞的生长无毒性影响。脱细胞基质材料在动物体内植入后早期有较多炎性细胞浸润,到实验观察的后期无明显炎性细胞浸润,脱细胞基质内可见成纤维细胞。另外,脱细胞基质材料对周边组织未产生毒性作用,伤口Ⅰ期愈合。同时组织学切片显示：支架材料与周边组织相容性好,未产生排斥反应。说明脱细胞基质材料在动物体内具有很好的生物相容性。 关键词：血管支架;生物相容性;生物材料;脱细胞血管基质;相容性 doi:10.3969/j.issn.1673-8225.2010.16.010     

Metacognition in schizophrenia: Correlates and stability of deficits in theory of mind and self-reflectivity

Research suggests that many with schizophrenia experience a range of deficits in metacognition including difficulties recognizing the emotions and intentions of others as well as reflecting upon and questioning their own thinking. Unclear, however, is the extent to which these deficits are stable over time, how closely related they are to one another and whether their associations with core aspects of the disorder such as disorganization symptoms are stable over time. To explore this issue, we administered three assessments of Theory of Mind (ToM), the Beck Cognitive Insight Scale (BCIS), and the Positive and Negative Syndrome Scale at baseline and 6 months to 36 participants with schizophrenia. Correlations revealed the ToM and BCIS scores were stable across the two test administrations and that the ToM tests were closely linked to each other but not to the BCIS. Poorer baseline performance on the ToM tests and the Self-Certainty scale of the BCIS were linked to greater cognitive symptoms at baseline and follow-up, while greater Self-Reflectivity on the BCIS was linked to greater levels of emotional distress at both baseline and 6-month follow-up. Results are consistent with assertions that deficits in metacognition are a stable feature of schizophrenia.