痫性放电实现异体间转道并致痫动物的研究报道

目的观察青霉素癫痫模型痫性放电能否被引导电极转道至异体大鼠脑内并致痫。方法实验大鼠海马局部注射青霉素建立癫痫模型,通过引导电极拟将痫性放电导入异体大鼠同侧海马,观察实验大鼠的行为学、脑电图变化。结果致痫组、痫能导出组12只大鼠全部点燃,痫能导入组6只大鼠亦出现痫性发作,痫能导出组痫性发作时程缩短,致痫组、痫能导出组、痫能导入组大鼠脑电图均可记录到痫性波;对照组、电极组无痫性发作。结论实验性痫性放电可通过引导电极在异体大鼠脑组织间传导,脑内痫性放电有可能被电极导出。     

视频脑电图监测对脑瘫合并癫痫发作性患儿的早期诊断及预后价值

目的分析视频脑电图监测对脑瘫合并癫痫发作性患儿的早期诊断及预后价值。方法回顾性分析62例脑瘫合并癫痫发作性患儿的临床资料,作为观察组;另选同期的62例脑瘫合并非癫痫发作性患儿作为对照组;所选患儿均采取视频脑电图监测,比较观察组和对照组患儿视频脑电图的痫样放电发生率,比较观察组与对照组患儿视频脑电图的同步节律放电发生率,并分析脑瘫合并癫痫发作性与视频脑电图异常放电的关系。结果观察组患儿视频脑电图的痫样放电发生率为79.03%,大于对照组(33.87%),差异具有统计学意义(P0.05);观察组患儿视频脑电图的同步节律放电发生率为66.13%,大于对照组的4.84%;差异具有统计学意义(P0.05);观察组视频脑电图显示发作期痫样放电发生率为64.52%,发作间期痫样放电发生率为41.94%,差异具有统计学意义(P0.05)。结论视频脑电图监测可显著提高脑瘫合并癫痫发作性患儿的痫样放电检出率,在临床发作的同时显示脑电图,分析其特征性变化,均有利于脑瘫合并癫痫发作性的早期诊断及预后评估。     

幼年大鼠癫痫持续状态后电生理和行为学的研究

目的观察幼年大鼠发生癫痫持续状态(status epilepticus,SE)后的电生理和行为学改变,并判断其表现是否符合Lennox-Gastaut综合征(Lennox-Gastaut syndrome,LGS)的电临床特点。方法 3周龄雄性Wistar大鼠50只,随机分为模型组(n=30)与对照组(n=20),模型组幼鼠腹腔注射氯化锂-匹鲁卡品诱发SE,对照组幼鼠用等量生理盐水代替匹鲁卡品。1周后,采用改进的颅内电极方法进行2周长程视频脑电监测,观察分析幼鼠脑电图背景活动、癫痫放电、癫痫发作及其相应症状学改变情况。脑电监测结束后,进行Morris水迷宫认知功能测试,观察记录幼鼠在水迷宫中寻找水下平台的潜伏期及游泳距离。结果共26只模型组幼鼠成功诱发SE并存活,对照组(n=20)表现正常。长程视频脑电监测期间,对照组幼鼠脑电背景活动正常,未见癫痫发作。模型组幼鼠发作间期癫痫放电显著增多,睡眠期可见弥漫性慢棘-慢波活动;共监测到3种癫痫发作类型:不典型失神发作、强直发作和肌阵挛发作。在水迷宫测试中,模型组幼鼠的潜伏期及游泳距离明显长于对照组(P0.05)。结论模型组幼鼠在发生SE后出现多种类型癫痫发作、异常脑电图表现及认知功能减退,该动物模型的电临床表现符合LGS的临床特征。     

尼莫地平对戊四氮慢性点燃癫痫大鼠行为及脑电图的影响

目的:探讨尼莫地平(nimodipine,NIM)对戊四氮(Pentylenetetrazole,PTZ)慢性点燃癫痫大鼠行为及脑电图的影响。方法:64只动物随机分为正常对照组(A组)、PTZ单纯致痫组(B组)、NIM1.25mg/kg干预组(C组)、NIM2.5mg/kg干预组(D组),通过腹腔注射PTZ建立慢性癫痫动物模型,观察大鼠行为、脑电图及海马组织学改变。结果:腹腔注射NIM可以明显抑制大鼠的痫性放电,减小其痫性发作级别,与正常对照组相比重型发作率明显降低(P<0.01),此外,NIM还能减轻海马的损伤程度。结论:NIM在PTZ慢性点燃癫痫大鼠模型中不仅具有抗惊厥作用,还能减轻海马的损伤程度。     

构建海人酸急性致痫小鼠模型

目的 建立海人酸诱导的小鼠急性癫痫模型,并探讨其特点.方法 实验取健康雄性昆明小鼠99只,随机分为生理盐水组(n=33)和致痫组(n=66),痫组腹腔注射海人酸10mg/kg,生理盐水组腹腔注射生理盐水35μl/g.注射后连续5h观察小鼠是否有痫性发作并分级.当小鼠持续痫性发作达1h时给予地西泮4mg/kg腹腔注射,对照组3只和致痫组9只同时描记脑电图,并取脑切片后苏木精-伊红染色法观察海马各区病理学改变.结果 ①行为学表现,模型组小鼠注射海人酸后可出现湿狗样抖动、头面部肌肉阵挛、肢体阵挛及全面强直阵挛发作.生理盐水对照组未见癫痫发作.②脑电图表现,癫痫持续状态小鼠表现为持续性节律性棘波、棘慢波或高波幅慢波.③病理学研究,双侧海马均可出现神经元变性,以CA1和门区为主.结论 腹腔注射海人酸致痫小鼠急性模型同相应的大鼠模型一样,具有制作简便、痫性发作潜伏期短、致痫率高等特点,其所产生的急性癫痫模型具有与人类颞叶癫痫相似的行为、脑电图与神经病理改变.     

青霉素与海人酸癫痫模型的比较

目的 通过用青霉素、海人酸2种药物制作癫痫模型,探讨2种致痫剂的作用特点及应用条件.方法 取健康雄性昆明小鼠90只,分为3组,对照组(n=10)、青霉素致痫组(n=40)和海人酸致痫组(n=40),青霉素致痫组腹腔注射青霉素7×106 U/kg,海人酸致痫组腹腔注射海人酸10mg/kg,生理盐水组腹腔注射生理盐水35 μL/g.注射后连续5 h观察小鼠是否有痫性发作并分级,进行脑电图描记.结果 空白对照组无痫性发作,两模型组均出现痫性发作,海人酸致痫组与青霉素致痫组按Racine分级0～Ⅴ级各级之间无明显差异(P＞0.05),小鼠出现癫痫的潜伏期海人酸致痫组比青霉素致痫组短(P<0.05),而且海人酸致痫组比青霉素致痫组死亡率低(P<0.05).结论 腹腔注射海人酸所致的动物模型具有与人类颞叶癫痫极为相似的癫痫发作行为学、脑电图特征,是理想的模拟人类颞叶癫痫的动物模型.     

损伤性癫痫模型大鼠海马和额叶突触蛋白的动态表达

目的 通过观察FeCl2皮层注射致损伤性癫痫(PTE)模型大鼠海马、额叶突触蛋白P38的表达变化.探讨突触可塑性在PTE发病机制中的作用. 方法 健康成年雄性SD大鼠采用随机数字表法分为正常对照组(n=5)、假手术组(n=12)、模型组(n=20).采取立体定向皮层注射FeCl2(0.1 moL/L,10μL)建立PTE模型,假手术组注入等量生理盐水,正常对照组不做处理.观察各组大鼠EEG的变化并应用免疫组织化学方法 检测大鼠造模后不同时间点(1 h、7 d、14 d、30 d)海马、额叶P38的表达. 结果大多数模型组大鼠在注射FeCl2后不久记录到癫痫样放电;与假手术组比较,模型组不同时间点右侧额叶P38表达减少.差异有统计学意义(P<0.05);与正常对照组和假手术组比较,致痫1h后,CA3区多形层、辐射层、腔隙层和齿状回(DG)分子层P38表达均无明显变化,7 d后P38表达增加,维持到30d,差异均有统计学意义(P<0.05). 结论 与突触蛋白P38表达相关的突触可塑性变化在PTE的发病机制中可能起重要作用.     

MicroRNA-132拮抗剂两种不同注射方式在匹罗卡品诱导的幼年大鼠癫痫急性期的差异

目的探讨microRNA-132拮抗剂对氯化锂-匹罗卡品诱导的幼年SD大鼠内侧颞叶癫痫(mesial temporal lobe epilepsy,MTLE)模型急性期的影响。方法实验分为4组:microRNA-132拮抗剂侧脑室置管组;microRNA-132拮抗剂阴性对照置管组;microRNA-132拮抗剂直接注射组;microRNA-132拮抗剂阴性对照直接侧脑室注射组;实验各组药物干预在造模前24 h进行。利用氯化锂-匹罗卡品建立SD大鼠MTLE模型,通过行为学观察各组大鼠癫痫持续状态发作潜伏时长,Racine评分观察各组大鼠抽搐发作的严重程度,脑电图监测癫痫样放电的频率及波幅,并统计实验各组大鼠诱导SE的成功率及24 h后的死亡率。结果在实验各组中,建模成功率无显著性差异,microRNA-132拮抗剂直接注射组幼年SD大鼠造模以后达到SE的潜伏时较其阴性对照组明显延长、癫痫发作的Racine评分明显下降、脑电图结果显示痫样放电的幅度及频率显著下降,死亡率降低,结果有统计学意义(P0.05)。MicroRNA-132拮抗剂侧脑室置管注射组与microRNA-132拮抗剂侧脑室直接注射组结果无统计学差异(P0.05)。结论 microRNA-132拮抗剂预处理SD大鼠能明显延长氯化锂-匹罗卡品诱导的SD幼年大鼠癫痫发作的潜伏期,减轻急性期抽搐严重程度及大脑样痫放电,降低大鼠癫痫持续状态后的死亡率。提示microRNA-132拮抗剂对氯化锂-匹罗卡品诱导的幼年SD大鼠MTLE的发生具有抑制作用,抑制microRNA-132有可能成为癫痫持续状态药物治疗的潜在靶点和新方向。     

褪黑素对FeCl_3致痫大鼠行为及脑电图的影响

目的:研究褪黑素(melatonin,MT)对FeCl_3致痫动物模型脑电图和行为改变的影响。方法:通过皮质内注射FeCl_3制备癫痫动物模型。32只雄性SD大鼠分为4组,即对照组、MT 0.03mmol·kg~(-1)组、MT 0.1mmol·kg~(-1) 组,MT 0.3mmol·kg~(-1)组,每组8只动物。观察大鼠行为表现及EEG改变。结果:0.3 mmol·kg~(-1)组可延长痫样放电的潜伏期,减少痫性放电总时间及长程发作次数、降低痫性行为评分。结论:褪黑素可抑制铁剂诱导的痫性活动和痫性放电。     

神经元突触可塑性在损伤性癫痫发病中的作用：立体定向Fecl2额叶皮质注射建立损伤性癫痫大鼠模型

目的 通过动态观察损伤性癫痫(PTE)模型大鼠皮层或海马超微结构变化,探讨神经元突触可塑性在PTE发病机制中的作用。方法　健康成年雄性SD大鼠采用随机数字表法分为正常对照组(n＝5)、假手术组(n＝12)、模型组(n＝20),采取立体定向皮层注射FeCl2(0.1 moL/L,10 μL)建立PTE模型,假手术组注入等量生理盐水,在不同时间点（1h、7d、14d、30d）取脑。电子显微镜下观察超微结构。结果　1. 大多数模型组大鼠在注射FeCl2后不久记录到癫痫样放电,死亡2只。2.电镜下FeCl2致痫组右额叶神经元胞体浓缩变性、粗面内质网增生、内质网的核糖体脱落、胶质细胞可见有空泡变性。第7天,右额叶细胞见吞噬颗粒,染色质边聚,可见早期细胞凋亡。第14天,右额叶细胞死亡,结构紊乱,胶质细胞核分裂像;大鼠海马神经毡内突触明显增多,多见Ⅰ型突触。第30天,大鼠海马,可见Ⅱ型突触,较多髓鞘。结论　神经元突触类型变化尤其是兴奋性突触增多在PTE的发病机制中起重要作用。     

急性氯化铁癫痫模型大鼠美解眠诱发放电的实验研究

目的 探讨急性氯化铁癫痫模型大鼠在发作间期美解眠诱发试验中的放电特征.方法 给予SD大鼠头颅额、顶、枕部铺设硬膜外电极6枚,用立体定向方法在大鼠感觉运动皮质区注入氯化铁溶液,建立急性癫痫模型,记录脑电24小时,观察在发作间期给予急性氯化铁癫痫模型大鼠腹腔注射美解眠后诱发癫痫发作的脑电情况.结果 美解眠诱发试验中,出现两种不同类型的癫痫发作期脑电,其中一种与急性氯化铁模型的发作期放电相同,另一种与美解眠自身所致癫痫的发作期放电相似.结论 急性氯化铁癫痫模型大鼠在美解眠诱发试验中能够产生原有癫痫发作,但是假阳性率较高;氯化铁致痫大鼠对美解眠的反应性较正常大鼠高.     

幼鼠和成年大鼠急性氯化铁癫痫模型的对比研究

目的 建立幼鼠和成年大鼠氯化铁癫痫模型,观察行为学,脑电图(EEG)和病理改变,对比分析两者在发作级别,发作次数,发作定位以及病理改变上的差异性.方法 将实验大鼠按鼠龄分幼年组(3周)和成年组(3个月).立体定向注射同剂量氯化铁(FeCl3)于大鼠左侧感觉运动皮层区.颅骨埋植不锈钢螺丝电极后,连续6小时行为学观察,EEG纪录,组织学观察.结果 在成功致痫的模型中,两组均表现较高级别的行为发作(Ⅳ级以上),成年组的模型成功率达80%,但幼年组的模型成功率较低,仅60%.两组大鼠EEG表现出相似的癫痫波形,但成年组在波幅上高于幼年组.同时在发作总时间和发作次数上,成年组均高于幼年组,结果具有显著差异性(P＜0.01).在癫痫发作的定位上,两组的分布相似.另外两组均有亚临床发作,以幼年组为显著(40%).病理改变两实验之间无明显差别.结论 成年大鼠氯化铁癫痫模型行为及EEG表现较稳定,而幼年大鼠氯化铁癫痫模型稳定性较差.     

颅内电极埋藏在枕叶癫痫中的作用

目的探讨运用颅内电极埋藏进行视频脑电图监测在定位困难的枕叶癫痫中的作用。方法通过对9例枕叶癫痫但定侧定位困难的患者,向颅内可疑部位植入硬膜下条状电极,进行视频脑电图监测,记录发作间期及发作期脑电图变化,确定癫痫病灶起始区。通过手术切除致痫灶。结果本组9例埋藏时间为3～9d,平均5d,均记录到间歇期痫样放电及发作期脑电图情况。行枕叶局部皮层切除6例及枕叶切除3例。术后按照Engel评分,I级7例,II级2例。所有病例均未出现埋藏电极引起的并发症。结论在致痫灶定位困难的顽固性枕叶癫痫中,采用颅内电极埋藏进行脑电图监测,可以精确定位致痫灶,从而提高癫痫的治愈率。     

青霉素致(癎)大鼠海马细胞内Ca2+浓度变化与神经元单位放电的关系

目的 探讨癫(癎)大鼠海马细胞内Ca2+浓度变化和神经元单位放电的关系.方法36只Wistar雄性健康大鼠分为3组.癫(癎)模型组用青霉素钠600万U/kg腹腔注射制作成癫(癎)模型;癫(癎)预处理组在制作模型前用盐酸氟桂利嗪按20 mg/kg间隔12 h灌胃2次,第2次给药后2 h用青霉素钠600万U/kg腹腔注射制作模型.3组实验大鼠均在记录单位放电后处死,取双侧海马应用新一代钙荧光指示剂Fluo-3-AM通过流式细胞分析仪测定细胞内Ca2+荧光浓度.结果 正常对照组大鼠共记录到24个单位海马神经元放电,海马细胞内Ca2+平均荧光浓度为37.12±5.08;癫(癎)模型组共记录到78个单位海马神经元放电,海马细胞内Ca2+平均荧光浓度为52.04±6.75;癫(癎)预处理组共记录到47个单位海马神经元放电,海马细胞内Ca2+平均荧光浓度为37.79±3.09.Ca2+平均荧光值和海马单位放电个数相关系数r=0.737,P＜0.01.结论 青霉素致病大鼠海马细胞内Ca2+浓度变化与海马神经元单位放电呈正相关关系.

Abstract：

Objective To investigate the relationship between changes in Ca2+ concentration and unit discharge in the hippocampal neurons of epileptic rats.Methods Thirty-six healthy male Wistar rats were divided into three groups. Rats of the epilepsy group received an intraperitoneal injection of benzylpenicllin sodium at 6 million unit/kg; Rats of the epilepsy pretreatment group received 20 mg/kg of flunarizine at 14 and 2 hours before injection of benzylponicllin;the control group of rats received an intraperitoneal injection of normal saline.Three groups of rats were euthanized after recording of the unit discharge,and bilateral hippocampal cells were obtained to measure the cellular concentration of Ca2+ fluorescence using the new generation Ca2+ fluorescent dye Fluo-3-AM and flow cytometry.Results (1) A total of 24 units of hippocampal discharges were recorded in the control group.The average concentration Ca2+ fluorescence of the hippocampal neurons was 37.12±5.08.(2)A total of 78 units of hippocampal discharges were recorded in the epilepsy group.The average concentration of Ca2+fluorescence of the hippocampal neurons was 52.04±6.75.(3)A total of 47 units of hippocampal discharges were recorded in the epilepsy pretreatment group. The average concentration of Ca2+ fluorescence of the hippocampal neurons was 37.79±3.09.The correlation coefficient between the average concentration of Ca2+ fluorescence of the hippocampal neurons and the number of units of hippocampal discharge ( r=0.737,P＜0.01).Conclusion There is a positive correlation between changes in average concentration of Ca2+ fluorescence of the hippocampal neurons and the numbers of units of hippocampal discharge.     

Value of long-term electroencephalogram in diagnosing epilepsy

BACKGROUND: Routine electroencephalogram (EEG) usually cannot accurately reflect the discharge of epileptic patients due to the short examination, and long-term EEG can make up the shortcoming. OBJECTIVE: To comparatively analyze the long-term EEG of epileptic and non-epileptic patients, and investigate the values of long-term EEG in the diagnosis and differential diagnosis of epilepsy. DESIGN: A case-controlled study. SETTING: Ningjin County People's Hospital. PARTICIPANTS: Totally 122 patients with epilepsy (epilepsy group) were selected from the EEG room of Ningjin County People's Hospital from January 2000 to December 2006, including 76 males and 44 females, 7 months to 78 years of age, the disease course ranged from 7 days to 7.5 years, and they all according with the standards for epilepsy set by the International Association for Epilepsy in 1997. Meanwhile, 118 patients with non-epileptic paroxysmal diseases were selected as the control group, including 71 males and 47 females, 2.5–87 years of age, the disease course ranged from 3 days to 7.5 years. Informed contents were obtained from all the subjects. METHODS: OXFORD GATE WAY 2000 16-lead portable EEG recorder was used for 24-hour electroencephalographic procedure. The patients could move normally during the monitoring, their activities, sleeping conditions, time and manifestations of seizures were recorded in details. In the next day, EEG at wake was recorded for 10 minutes, followed by 3-minute hyperventilation and open/close eye induction test, the phases of non-rapid eye movement (Ⅰ–Ⅳ) and rapid eye movement were performed using EEG at sleep according to the international EEG standard. The abnormal rates of EEG epileptic discharge at wake and sleep at different sites were calculated. MAIN OUTCOME MEASURES: Abnormal rate of long-term EEG at wake and sleep in both groups; Epileptic discharge at different sleeping phases in both groups; Abnormal rates of EEG epileptic discharge at wake and sleep at different sites in the epilepsy group. RESULTS: All the 122 patients with epilepsy and 118 patients with paroxysmal diseases were involved in the final analysis of results. ① Comparison of abnormal rate of long-term EEG at wake and sleep: In the epilepsy group, the abnormal rate of EEG at wake was obviously lower than that at sleep (68%, 91%, P < 0.01). In the control group, the abnormal rate of EEG at wake and sleep had no obvious difference (P > 0.05). ② Results of epileptic discharge at different sleeping phases: In the epilepsy group, the epileptic discharge occurred at Ⅰ–Ⅱ phases of sleep cycle in 88.1%, and at Ⅲ–Ⅳ in 11.9%; In the control group, the epileptic discharge occurred at Ⅰ–Ⅱ phases of sleep cycle in 91.7%, and at Ⅲ–Ⅳ phases in 8.3% (1/12). ③ Comparison of the abnormal rates of EEG epileptic discharge at wake and sleep at different sites in the epilepsy group: The abnormal rates of epileptic discharge at frontal lobe and temporal lobe at sleep were obviously higher than those at wake (21.3%, 24.6%; 10.7%, 11.7%, P < 0.01), while there were no obvious differences at wake and sleep at occipital lobe, parietal lobe (P > 0.05). CONCLUSION: Long-term EEG has great importance in the diagnosis and differential diagnosis of epilepsy, especially that it increases the detective rate of discharge by several cycles of sleep derivation. This method also provides important reference for the allocation of epileptic focus.     

颅内电极埋藏在Lennox-Gastaut综合症中的应用(附10例临床分析)

目的探讨颅内电极埋藏术后进行视频脑电图评估在癫痫外科手术致痫灶定位困难的Lennox-Gastaut综合症中的使用。方法收集10例Lennox-Gastaut综合症致痫灶定位困难的患者,向颅内硬膜下植入条状电极,术后进行视频脑电图评估,记录发作间歇期及发作期脑电图变化,确定癫痫病灶的起始区,通过手术方式切除致痫灶。结果本组10例患者埋藏时间为2～7天,平均4天,均记录到间歇期及发作期脑电图情况。根据脑电图结果,行脑叶切除及胼胝体切开。术后按照Engel评分I级4例,II级2例,III级2例,IV级2例。所有病例均未出现埋藏电极引起的严重并发症。结论在致痫灶定位困难的Lennox-Gastaut综合症中,采用颅内电极埋藏进行视频脑电图检测,可以较准确定位主要致痫灶,从而提高Lennox-Gastaut综合症外科治疗有效率。     

机器人辅助无框架定位颅内电极埋植(附19例分析)

目的 探讨机器人辅助无框架定位颅内电极埋植后行长程视频脑电图(VEEG)监测定位癫(癎)起源灶的方法及效果.方法 对19例顽固性癫(癎)病人采用机器人辅助无框架定位进行颅内深部电极埋植.其中15例术前无法定位致(癎)灶者均明确癫(癎)起源灶所在脑叶,4例术前检查提示为单侧颞叶(癎)性放电者证实为颢叶起源灶.根据监测结果,采用开颅手术3例,立体定向脑内致(癎)灶毁损术10例,伽玛刀治疗6例.结果 术后疗效按Engel分级:Ⅰ～Ⅱ级7例,Ⅲ级6例,Ⅳ级5例,Ⅴ级1例.结论 机器人辅助无框架定位颅内电极埋植定位癫(癎)起源灶的方法具有简单、微创等特点,适用于头皮EEG无法定位者,尤其是对颞叶癫(癎)的起源灶定侧与定位具有重要价值.     

Periictal SPECT localization verified by simultaneous intracranial EEG

PURPOSE: We investigated whether blood-flow changes measured by ictal or immediate postictal single photon emission computed tomography (SPECT) reflect with accuracy the actual location of ictal discharge as measured by simultaneous intracranial EEG. In addition, we evaluated the reliability of ictal SPECT obtained with implanted electrodes by comparing results with those of ictal SPECT performed during scalp EEG monitoring in selected patients. METHODS: Eleven patients with intractable partial epilepsy who had both ictal and interictal SPECT scans during invasive EEG monitoring were studied. We analyzed perfusion differences based on registration, normalization, and subtraction of periictal and interictal SPECT images. SPECT results were interpreted in relation to location and evolution of ictal EEG change, as reflected by simultaneous intracranial EEG. In five patients, we also compared ictal SPECT results that were obtained during both scalp and intracranial EEG monitoring. RESULTS: In 10 of 11 patients, localized increases or decreases in blood flow or both were identified in regions of ongoing or prior seizure discharge, respectively, at the time of SPECT brain perfusion. In one patient, SPECT localization could not be verified by the available electrode array. CONCLUSIONS: Localization of ictal discharge during or before SPECT injection accurately determines increase or decrease in perfusion, respectively, and both are of equal validity in reflecting the region of epileptic discharge. SPECT perfusion changes can be reliably obtained during intracranial monitoring.