外伤性癫痫灶中泛素蛋白酶体系的形态学改变

目的检测泛素(ubiquitin,Ub)和泛素激活酶(ubiquitin-activating enzyme1,UbE1)在外伤性癫痫灶中的表达,探索其对鉴定外伤性癫痫的应用价值。方法收集脑外伤后癫痫灶皮质样本15例(外伤性癫痫组)、非脑外伤后癫痫灶皮质样本15例(非外伤性癫痫组),交通事故死亡正常脑皮质样本15例(非癫痫组),通过HE常规染色观察各组形态学改变,用免疫组化染色检测3组中Ub和UbE1的表达情况。结果常规HE染色下,与非癫痫组相比,外伤性癫痫组和非外伤性癫痫组出现神经元数量减少、神经元变性等形态学改变,外伤性癫痫改变更明显。Ub和UbE1主要表达在癫痫灶神经元细胞核、细胞质内,细胞外未见表达。图像分析显示两指标的表达量为外伤性癫痫组非外伤性癫痫组非癫痫组,差别具有统计学意义(P0.05)。结论正常神经元数量减少、神经元变性是癫痫灶的主要病理改变之一,外伤性癫痫神经元病理改变更明显。与HE染色比较,Ub和UbE1免疫组化染色更灵敏,有助于鉴别外伤性癫痫和非外伤性癫痫。     

外伤性癫痫患者癫痫灶中GABABR1和GluR1的表达

目的探讨外伤性癫痫患者颞叶癫痫灶组织γ-氨基丁酸B受体亚单位（gamma-aminobutyric acid B receptorsubunit,GABABR1）和谷氨酸受体亚单位（glutamic acid receptor subunit,GluR1）的表达变化,以期为阐释外伤性癫痫发生机制和法医学鉴定提供依据。方法收集15例外伤性癫痫患者脑皮质（外伤性癫痫组）1、5例非外性癫痫患者脑皮质（非外伤性癫痫组）、15例交通事故死亡者脑皮质（交通事故死亡对照组）,运用荧光免疫组化技术和RT-PCR技术观察GABABR1和GluR1的表达水平。结果外伤性癫痫组GABAB R1表达水平明显低于对照组,但高于非外伤性癫痫组;外伤性癫痫组GluR1表达水平明显高于对照组,但低于非外伤性癫痫组。结论 GABABR1和GluR1的表达水平的变化在外伤性癫痫病的发病过程中起着一定的作用,可以作为区分外伤性癫痫与非外伤性癫痫的病变化指标之一。     

外伤性癫痫灶中泛素与泛素激活酶的定量分析

目的 定量测定人体外伤性癫痫灶中泛素(ubiquitin,Ub)和泛素激活酶(ubiquitin-activating enzyme,UbE1)的表达,研究泛素蛋白酶体系在外伤性癫痫(post-traumatic epilepsy,PTE)发病机制中所起的作用,探索其在PTE鉴定中的应用价值. 方法 分别收集颅脑外伤致癫痫患者脑癫痫灶组织(外伤性癫痫组)、非颅脑外伤性癫痫灶组织(非外伤性癫痫组)和交通事故死亡者正常脑组织(非癫痫组)各15例,应用RT-PCR、Western印迹技术检测3组中Ub和UbE1的含量,并对所得数据进行统计学分析. 结果 RTPCR、Western印迹技术测定显示Ub和UbE1的mRNA和蛋白质在3组中含量为外伤性癫痫组>非外伤性癫痫组>非癫痫组,两两比较,组间差异具有统计学意义(P<0.05).结论 Ub和UbE1在癫痫灶组织中表达增加,且在外伤性癫痫灶组织中增加最为明显.推断泛素蛋白酶体系的活化是PTE神经元病理改变的重要机制之一.     

外伤性癫痫灶组织的基因表达谱分析

目的比较外伤性和非外伤性癫痫灶组织的基因表达谱,探索外伤性癫痫的发病机制,为相关鉴定提供依据。方法收集外伤性和非外伤性癫痫灶组织样本各15例。分别提取总RNA,经逆转录标记不同的探针与人类cDNA表达谱芯片杂交,获得扫描图像,分析信号的强度和比值,对实验数据按照RNA微阵列分析法标准化,对差异表达基因进行聚类分析。结果两组样本中共有超过2 990条基因明显表达（表达超过背景2～5倍以上）。以30%样本中的表达调节水平大于1.5倍作为差异基因筛选标准,与非外伤性癫痫组比较,外伤性癫痫组样本中存在显著性表达差异的基因有192个,其中表达上调的有115个,表达下调的有77个。结论采用基因表达谱芯片技术可以有效地识别外伤性癫痫和非外伤性癫痫的差异表达基因,并可用于外伤性癫痫发病机制和分子分型的研究。     

外伤性癫痫病灶中caspase-10的表达

【摘要】目的观察凋亡基因caspase-10存外伤性癫痫病灶中的表达变化,探讨其作用机制。方法收集外伤性、非外性癫痫患者手术切除的恼皮质及交通事故死亡者脑皮质各15例,运用RT-PCR技术观察比较各实验组凋亡基因caspase-10表达水平,采用荧光免疫组织化学染色技术观察各实验组凋亡基因对应蛋白的表达情况。结果外伤性癫痫组（：aspase-10基冈和蛋白表达水平比非外伤性癫痫组和交通事故死亡组高,差异有统计学意义（P〈0．05）;非外伤性癫痫组caspase-10基因和蛋白表达水平亦显著高于交通事故死亡组（P〈0．05）。结论凋亡基因caspase－10能促进神经元凋亡,对外伤性癫痫的发病过程有促进作用。     

Bcl-2蛋白在外伤性癫痫病灶中的表达

目的观察Bcl-2蛋白在外伤性癫痫病灶中的表达变化,探讨其作用机制。方法收集外伤性癫痫、非外性癫痫患者及交通事故死亡者脑皮质各15例,采用免疫组织化学和荧光免疫组织化学技术,观察各实验组Bcl-2蛋白表达水平,测定阳性表达的灰度值,数据采用SPSS 12.0软件进行统计分析。结果外伤性及非外伤性癫痫组Bcl-2蛋白表达水平均高于交通事故死亡组（P〈0.05）;而非外伤性癫痫组表达明显高于外伤性癫痫组,差异亦具有显著统计学意义（P〈0.05）。结论 Bcl-2蛋白能抑制神经元凋亡,对外伤性癫痫的发病过程具有抑制作用。     

大鼠外伤性癫痫模型皮层SPP1蛋白的表达

目的检测分泌型焦磷酸蛋白1 (Secreted phosphoprotein 1,SPP1)在外伤性癫痫(post-traumaticepilepsy,PTE)大鼠皮层脑组织中的表达变化,探讨其与外伤性癫痫的关联。方法随机将18只大鼠分为正常对照组、生理盐水组和实验组。建立外伤性癫痫模型。采用免疫组化法检测GFAP蛋白,采用Western blot法检测SPP1蛋白。结果大鼠行为学平均评分为4分;EEG显示明显痫性放电;免疫组化结果显示,实验组大鼠脑组织GFAP阳性细胞与正常对照组和生理盐水组比较,显著增高(P <0.05);生理盐水组GFAP阳性细胞与正常对照组比较,差异不具有统计学意义(P> 0.05),以上结果综合表明成功建立了大鼠外伤性癫痫模型。Western blot结果显示,实验组SPP1蛋白表达量较正常对照组和生理盐水组显著增高(P <0.01);生理盐水组SPP1蛋白表达量较正常对照组不显著,差异无统计学意义(P> 0.05)。结论 SPP1在外伤性癫痫大鼠皮层脑组织中表达显著上调,外伤性癫痫的发生机制可能与SPP1的表达异常增高有关。     

颅脑外伤后假性癫痫的法医学鉴定

通过对2例颅脑外伤后假性癫痫的法医鉴定案例的分析,总结假性癫痫发作的临床特点及外伤后假性癫痫发作与外伤性癫痫发作的鉴别要点。其法医学鉴定应结合颅脑外伤的程度、部位、颅内有无后遗病灶及临床发作特点等综合分析。     

外伤性癫痫及其法医学评定进展

外伤性癫痫(post traumatic epilepsy,PTE)是指颅脑损伤后的癫痫性发作,影像学检查可见器质性病变,脑电图检查可见异常脑电波与颅脑损伤部位一致。外伤后癫痫发生率低,案例报道较少,患者易夸大病情,给法医学鉴定增加难度。本文就外伤性癫痫流行病学、发病机制、临床治疗、法医学鉴定等方面的现状进行综述。     

大鼠局灶性脑损伤后calpain1和calpain2的表达

目的探讨大鼠局灶性脑损伤后钙激活中性蛋白酶1和2（calpain1,calpain2）表达规律与损伤时间的相关性。方法健康成年雄性SD大鼠50只,体重200～250g。随机分为10组,每组5只,其中实验组8组,对照组和假手术组各1组。应用免疫组化染色法,检测大鼠脑损伤后不同时程损伤周边区calpain1和calpain2的表达变化。结果对照组及假手术组均有capain1和calpain2表达。伤后15min,损伤周边区calpain1/calpain2阳性染色开始增强,伤后12h达高峰,24h逐渐减弱,5d阳性表达增强达另一高峰,随后逐渐减弱,至15d接近对照组水平。除伤后15d组外,其余实验组与对照组比较,calpain1和calpain2阳性反应物面积百分比及平均光密度值差异均具有统计学意义（P〈0.05）。结论在正常大鼠脑组织中calpain1和calpain2均有表达,局灶性脑损伤后,损伤周边区calpain1和calpain2的表达随时间呈较典型的双峰变化。     

外伤性癫痫的法医学鉴定21例分析

为了探讨颅脑损伤后癫痫的法医学鉴定要点,对 21例颅脑损伤后癫痫出现发作者进行了现病史、既往史调查,并结合临床脑电图（ EEG）及 X线片、 CT片、 MRI片等影像学资料。结果发现外伤后癫痫常继发于严重的颅脑损伤,多以晚期发作为主,其发作性质决定于颅脑损伤的部位和程度,头皮 EEG和 24小时动态 EEG证实有异常癫痫样波存在,影像学检查有助于定性分析。结果提示：外伤性癫痫法医学鉴定必须以掌握外伤史及既往史为基础,结合 EEG、 CT、 MRI等检查方可作出正确鉴定。     

The expression of excitatory amino acid transporter 2 in traumatic brain injury

It is well recognized that glutamate is the major excitatory neurotransmitter, which is removed from the synaptic cleft by excitatory amino acid transporter 2 (EAAT2) located on the perisynaptic astrocytes and that neuronal death has been associated with an increased extracellular glutamate concentration. In this study, we have immunohistochemically demonstrated the expression of EAAT2 protein in the human brain after traumatic brain injury (TBI). The EAAT2 expression patterns can be divided into three types: continuous and highly extensive staining (E); continuous but sporadic staining (M); and sporadic pattern staining (S). In six of the nine short survival cases studied (1 h to 1 day), continuous and highly extensive staining for EAAT2 (E type) was observed in the ipsilateral cerebral cortex. On the other hand, we were able to demonstrate weak staining (S and M types) in 5 of the 7 long survival cases (> or =1 day) and in 12 of the 14 very short survival cases (<1 h) studied. Similar findings were obtained in the contralateral cerebral cortex and also in the ipsilateral hippocampus. In addition, positive staining for glial fibrillary acidic protein was detected around the cerebral contusion, but the EAAT2-positive expression was not observed in the same region for all of the six short and long survival cases (> or =1 h) after TBI. These findings clearly showed the differences in EAAT2 expression in the cerebral cortex according to the survival time and severity of cerebral contusion after TBI. Therefore, we emphasized that EAAT2 might play an important role in contributing to extracellular glutamate concentrations and secondary brain injury after TBI.     

The expression of excitatory amino acid transporter 2 in traumatic brain injury

It is well recognized that glutamate is the major excitatory neurotransmitter, which is removed from the synaptic cleft by excitatory amino acid transporter 2 (EAAT2) located on the perisynaptic astrocytes and that neuronal death has been associated with an increased extracellular glutamate concentration. In this study, we have immunohistochemically demonstrated the expression of EAAT2 protein in the human brain after traumatic brain injury (TBI). The EAAT2 expression patterns can be divided into three types: continuous and highly extensive staining (E); continuous but sporadic staining (M); and sporadic pattern staining (S). In six of the nine short survival cases studied (1 h to 1 day), continuous and highly extensive staining for EAAT2 (E type) was observed in the ipsilateral cerebral cortex. On the other hand, we were able to demonstrate weak staining (S and M types) in 5 of the 7 long survival cases (≥1 day) and in 12 of the 14 very short survival cases (<1 h) studied. Similar findings were obtained in the contralateral cerebral cortex and also in the ipsilateral hippocampus. In addition, positive staining for glial fibrillary acidic protein was detected around the cerebral contusion, but the EAAT2-positive expression was not observed in the same region for all of the six short and long survival cases (≥1 h) after TBI. These findings clearly showed the differences in EAAT2 expression in the cerebral cortex according to the survival time and severity of cerebral contusion after TBI. Therefore, we emphasized that EAAT2 might play an important role in contributing to extracellular glutamate concentrations and secondary brain injury after TBI.     

外伤性癫痫法医学评定100例分析

目的探讨颅脑外伤后癫痫的法医学评定要点,提出外伤性癫痫的法医学评定依据。方法对100例颅脑损伤后癫痫患者门诊及住院病历,结合临床脑电图（EEG）结果、CT片、MRI片等影像学资料,进行了回顾性分析。结果依据GBl8667-2002《道路交通事故受伤人员伤残评定》标准有关颅脑损伤后癫痫发作类型、发作频率的相关条款规定,Ⅲ级伤残8人,Ⅴ级伤残10人,Ⅶ级伤残22人,Ⅸ级伤残40人,Ⅹ级伤残20人。误工期限、营养期限和护理期限评定结论时间长短差异较大,具有个体化特征。结论外伤后癫痫常继发于严重的颅脑损伤,其发作特点决定于颅脑外伤的部位和程度,外伤性癫痫法医学鉴定必须以外伤史及既往史为基础,结合电生理及影像学检查,综合癫痫发作类型、频率及药物控制情况作出正确的伤残等级及误工期限、护理期限、营养期限的鉴定结论。