A quantitative immunohistochemical study on the time-dependent course of acute inflammatory cellular response to human brain injury

The time-dependent inflammatory cell reaction in human cortical contusions has been investigated during the first 30 weeks after blunt head injury. Immunohistochemical staining was carried out using CD 15 for granulocytes and LCA, CD 3 and UCHL-1 for mononuclear leucocytes. In order to provide reliable data for a forensic wound age estimation, the intensity of the cellular reaction was evaluated with a quantitative image analysis system. CD 15-labelled granulocytes were detectable earliest 10 min after brain injury, whereas significantly increased numbers of mononuclear leucocytes occurred in cortical contusions after a postinfliction interval of at least 1.1 days (LCA), 2 days (CD 3) or 3.7 days (UCHL-1), respectively. Received: 15 June 1998 / Received in revised form: 6 October 1998     

The time course of the vascular response to human brain injury – an immunohistochemical study

In a total of 104 individuals who had sustained traumatic brain injury (TBI), the time-dependent vascular response was investigated at the injured cortical area during the first 30 weeks after the trauma. The immunohistochemical staining of the cerebral blood vessels was performed with antibodies against laminin, type IV collagen, tenascin, thrombomodulin and factor VIII associated antigen. Compared to the immunoreactivity in unaltered control tissue, a significantly increased vascular expression could be detected in cortical contusions after a postinfliction interval of at least 3 h for factor VIII, after 1.6 days for tenascin or after 6.8 days for thrombomodulin, whereas the immunostaining for laminin and type IV collagen was regularly positive even in the vascular endothelium of uninjured brain tissue. Received: 22 April 1999 / Received in revised form: 7 October 1999     

急性脑静脉闭塞脑损伤治疗时间窗初探:DWI与病理学对照实验研究

目的:探讨急性脑静脉闭塞模型脑实质损伤治疗时间窗的存在及其意义.方法:选择新西兰大白兔28只随机分为2组(实验组24只,对照组4只),实验组动物经一侧颈内静脉注入醋酸纤维素聚合物(CAP),分别于术后1、3、6、12、24和48 h行T1WI、T2WI和扩散加权成像(DWI)检查.各时间点MR扫描后取兔脑组织做胶质纤维酸性蛋白(GFAP)表达的免疫组化研究及电镜观察.结果:DWI、T2WI、GFAP的表达和电镜检查均能显示急性脑静脉闭塞模型脑实质损伤及其变化.DWI在术后1 h即能显示脑实质病变(ADC值下降),术后3 h DWI和T2WI均能显示病变;术后6 h前,DWI上有扩散异常的脑组织容积明显大于T2WI上异常高信号区的容积(t=13.69,P＜0.01);术后12、24和48 h病变区ADC值逐渐回升,T2WI上病变容积与DWI上的扩散异常区的容积比较,差异无显著性意义(t值分别为1.467、0.996和2.017,P＞0.05).术后1 h病变区GFAP阳性细胞增多,染色加深,胞体增大,突起增粗增长,术后3～6 h变化更明显,病理学改变以血管源性水肿为主,12 h后出现脑组织大量坏死.对照组未见上述各种异常表现.结论:DWI可准确评价急性脑静脉闭塞模型脑实质损伤程度,结合GFAP的表达,在探讨急性脑静脉闭塞脑损伤的治疗时间窗的存在及其意义中具有重要价值,在其发生发展过程中确实存在潜在的治疗时间窗.     

亚低温治疗重型颅脑损伤患者临床疗效分析及对患者血清GFAP、UCH-L1、NSE、CRP水平的影响

目的探讨亚低温治疗重型颅脑损伤患者临床疗效及对患者血清神经胶质纤维酸性蛋白(GFAP)、泛素羧基末端水解酶L1(UCH-L1)、神经元特异性烯醇化酶(NSE)、C反应蛋白(CRP)水平的影响。方法 106例重型颅脑损伤患者根据随机数字表法分为治疗组(n=53)和对照组(n=53)。对照组给予降低颅内压、脱水以及营养脑细胞等常规治疗,治疗组在对照组基础上结合亚低温治疗。对比分析两组入院时、治疗后1d和治疗后5d颅内压、血糖、血清GFAP、UCH-L1、NSE、CRP指标变化以及治疗6个月后预后情况。结果两组颅内压第1天、第5天较入院时明显降低(P0.05);治疗组治疗后1、5d颅内压明显低于对照组(P0.05);两组第1、第5天血糖水平显著低于入院时(P0.05);治疗组治疗后1、5d血糖水平显著低于同期对照组(P0.05);两组血清GFAP、UCH-L1水平治疗后1、5d较入院时显著降低(P0.05);治疗组血清GFAP、UCH-L1水平治疗后1、5d显著低于对照组(P0.05);两组NSE、CRP水平治疗前比较差异无统计学意义(P0.05);治疗组NSE、CRP治疗后1d和5d明显低于同期对照组(P0.05);治疗组良好率显著高于对照组(P0.05),病死率显著低于对照组(P0.05);而两组中残、重残及植物状态对比差异无统计学意义(P0.05)。结论亚低温治疗重型颅脑损伤患者临床疗效显著,降低血清GFAP、UCH-L1、NSE、CRP水平,改善患者预后,值得进一步推广应用。     

The expression of tumor necrosis factor-α in the rat brain after fluid percussive injury

To investigate the role of tumor necrosis factor-α (TNFα) after traumatic head injury in rats, moderate brain injury of 1000 mmHg was generated by an original fluid percussion injury device. TNFα levels in cerebrospinal fluid (CSF) gradually increased during the first 1 h, rose to a maximal elevation at 3 h and 6 h and returned to basal values by 24 h. Horseradish peroxidase tracer experiments revealed that primary microvascular damage appeared as early as 15 min after impact, but rapidly recovered and 1 h after impact secondary microvascular damage occurred in the hippocampus and parasagittal cortex. By immunoelectron microscopy, TNFα reactions were detected in the lysosomes of microglia accumulated at the impact site of the cortex 30 min after impact, and 1 h after impact these reactions were mainly detected at the glial cells (such as microglia and astrocytes) in the hippocampus and parasagittal cortex. Therefore the delayed microvascular damage observed in sites remote from the impact may be induced by TNFα which is synthesized mainly by glial cells. The present study suggests that TNFα conveyed from the microglial cells is one cofactor contributing to the fluid percussive brain edema formation after moderate brain injury. Received: 13 February 1997 / Received in revised form: 10 July 1997     

大鼠脊髓打击伤模型的建立及病理观察

目的 观察改良大鼠脊髓打击伤模型的病理变化,评价该模型的稳定性、重复性和一致性. 方法 应用改良重物坠落打击装置建立大鼠脊髓中、重度损伤模型,通过BBB评分、神经电生理监测、免疫组化及光电镜技术观察脊髓损伤(SCI)后两组大鼠功能及病理变化特点,以评价本实验模型的重复性及与损伤程度相关的一致性. 结果 脊髓损伤后两组大鼠的后肢运动功能均有不同程度恢复,运动和感觉诱发电位提示神经信号传导功能亦逐渐恢复,但中度损伤大鼠恢复明显较重度损伤大鼠好,组织病理及免疫组化观察显示脊髓损伤后有胶质瘢痕及囊腔形成,但重度损伤组的组织损伤程度较中度组重,且残留正常组织较少,电镜下观察显示脊髓损伤后损伤部位可见明显出血、水肿,中性粒细胞浸润,胶质细胞染色质边集.脊髓损伤后2周可见线粒体空泡化,轴突变性,周围水肿.脊髓损伤后8周可见髓鞘变性,胶原原纤维沉积. 结论 本实验模型能将不同打击力度造成的损伤区分开,且和行为学、神经电生理、病理学等结果吻合,说明此模型具有良好的稳定性、重复性和一致性.胶质瘢痕及囊腔的形成是脊髓损伤的标志性病理变化,GFAP或Vimentin染色可作为判断其界限的标志性分子.     

Course of glial immunoreactivity for vimentin, tenascin and alpha1-antichymotrypsin after traumatic injury to human brain

In a total of 104 individuals who had sustained traumatic brain injury (TBI), the course of glial immunoreactivity was investigated at the injured cortical area during the first 30 weeks after the trauma, in order to provide data for a forensic wound age estimation. Glial cells were stained with antibodies against the intermediate filament protein vimentin, the extracellular matrix protein tenascin and the serine protease inhibitor α1-antichymotrypsin (α1-ACT). Injury-induced glial staining reactions could be observed, at the earliest, after a post-infliction interval of 3.1 h for α1-ACT, 22 h for vimentin and 7 days for tenascin. Received: 21 August 2000 / Accepted: 21 November 2000     

GFAP重组蛋白表达纯化及其主动免疫对脊髓损伤的神经保护作用

目的 构建表达胶质原纤维酸性蛋白(glial fibillary acidic protein,GFAP)的质粒,诱导表达GFAP蛋白,纯化鉴定后主动免疫大鼠,并通过SCI大鼠行为学研究GFAP蛋白主动免疫在脊髓损伤(spinal cord injury,SCI)中的神经保护作用,为开展下一步研究工作做准备.方法 从人...     

The role of tumor necrosis factor-α in diffuse axonal injury following fluid-percussive brain injury in rats

The immunolocalization of tumor necrosis factor-α (TNFα) after diffuse axonal injury (DAI) is demonstrated using a midline fluid percussion rat model (moderate brain injury of 1000 mm Hg was generated) and the effects of TNFα on the axolemmal permeability using horseradish peroxidase as a tracer. In addition, the accumulation of β-amyloid precursor protein (β-APP) was investigated, which has recently been shown to be a reliable marker for the diagnosis of DAI in cases with fatal head injury. TNFα levels in brain tissues from the impact site and the cortex including the corpus callosum, gradually increased during the first 1 h, rose to a maximal elevation at 3 h, gradually decreased at 6 h and decreased further at 24 h. Horseradish peroxidase (HRP) tracer experiments revealed that primary axonal damage appeared as early as 15 min after impact but rapidly recovered and that 1 h after impact, secondary axonal damage occurred in the corpus callosum and the brain stem. By immunoelectron microscopy it was seen that β-APP accumulated in the axon from 1 h after impact demonstrating that there was functional axonal damage. TNFα reactions were detected in the lysosomes of microglia 30 min after impact and 1 h after impact these reactions were mainly detected in the glial cells (such as microglia, astrocytes and oligodendrocytes) in the corpus callosum and the brain stem. It is generally accepted that TNFα directly induces primary demyelination and oligodendrocyte apoptosis. Therefore, TNFα conveyed from the microglial cells is one cofactor contributing to the formation of the delayed axonal damage observed at these sites. The present study suggests that TNFα conveyed from the glial cells may contribute to the pathogenic mechanism of DAI formation following fluid percussive brain injury. Received: 27 April 1999 / Received in revised form: 26 July 1999     

脑星形细胞瘤MRI表现与GFAP表达的关系

目的 :探讨星形细胞瘤MRI表现与GFAP表达的关系。方法 :收集手术病理证实的脑星形细胞瘤 4 1例 ,分析其MRI表现。采用免疫组化方法标记肿瘤GFAP。对照GFAP表达程度与MRI表现的关系。结果 :星形细胞瘤GFAP表达强弱与病理级别相关 ,星形细胞瘤信号均匀性、边界清楚与否、瘤周水肿程度及增强程度与GFAP表达之间有统计学差异 ,但肿瘤部位及大小与GFAP表达没有相关性。结论 :星形细胞瘤MRI表现 ,特别是脑水肿、增强程度等征象 ,在一定程度上可以反映GFAP表达强弱。     

Myocardial injury due to lightning

The report deals with a 27-year-old male who was standing in a tent and was injured by lightning as it struck a tree about 1.5 m away. He immediately lost consciousness and exhibited ventricular fibrillation when the emergency physician arrived. A clinical picture of hypoxaemic brain damage emerged after initially successful resuscitation. Brain death was diagnosed on the fifth day after injury. The discrete external findings (remaining arborescent skin marks) contrasted markedly with the severe thermal damage to the pectoral muscle and cardiac musculature found during the autopsy. The histological cardiac findings indicated severe acute myocardial infarction affecting virtually all parts of the myocardium. Received: 3 February 1997 / Received in revised form: 7 May 1997     

联合检测血清S-100B蛋白、神经元特异性烯醇化酶、神经胶质纤维酸性蛋白对严重颅脑损伤诊断及预后的意义

目的探讨严重颅脑损伤后血清S-100B蛋白、神经元特异性烯醇化酶(NSE)、神经胶质纤维酸性蛋白(GFAP)水平变化及其临床意义。方法采用酶联免疫吸附法(ELISA)检测46例严重颅脑损伤患者伤后12、24h、3、7、14d血清S-100B蛋白、NSE、GFAP水平,其中12h～3d的平均值作为初期值结合格拉斯哥昏迷评分(GCS)分析;7～14d平均值作为后期值结合格拉斯哥预后评分(GOS)分析,并与峰值及平均值比较。结果 (1)严重颅脑损伤后血清S-100B蛋白、NSE、GFAP水平较对照组明显升高;(2)重型(GCS6～8分)与特重型颅脑损伤组(GCS 3～5分)相比,3种标记物均无显著性差异(P>0.05);(3)预后不良组(GOS 1～3分)标记物峰值、平均值及后期值均明显高于预后良好组(GOS 4～5分)(P<0.005),3个值均与预后呈负相关,后期值与预后的相关性最大。3种标记物中GFAP后期值差异尤为显著(P<0.001)。结论严重颅脑损伤后血清S-100B蛋白、NSE、GFAP水平升高,但不能作为评判重型颅脑损伤或特重型颅脑损伤的依据。3种标记物水平对评估重型颅脑损伤预后具有较高的特异性和敏感性,其中GFAP后期值是评估严重颅脑损伤预后的较好的指标。     

大鼠颅脑创伤后星形细胞早期反应特点

目的研究大鼠颅脑创伤后星形细胞的早期反应。方法用气体冲击导致大鼠局部脑挫伤,采用HE染色和透射电镜观察损伤后脑组织的病理变化,用胶质纤维酸性蛋白免疫组化染色检测反应性星形细胞。结果颅脑创伤后星形胶质细胞肿胀为一早期现象,反应性星形胶质细胞中胶质纤维酸性蛋白染色增强,伤后6小时,反应性星形细胞主要局限于伤区周围脑组织和伤侧海马,伤后24小时,星形细胞反应波及全脑,海马组织中尤为明显。结论颅脑创伤后星形细胞反应可能与早期胶质细胞应激反应有关,反应性星形细胞在继发性脑损伤病理过程中可能具有重要作用。     

Heat-stroke-induced cerebellar atrophy: clinical course, CT and MRI findings

We report the clinical course and CT and MRI findings in a case of heat-stroke-induced cerebellar atrophy. Although the cerebellar syndrome was severe concomitant with the onset of heat stroke, no abnormality was observed on brain CT in the first 2 weeks following the event. Cerebellar atrophy was first noted after 10 weeks on MRI; it was progressive during a 1-year follow-up. Received: 30 November 1995 Accepted: 1 March 1996     

大鼠脊髓缺血损伤后神经微丝及胶质纤维酸性蛋白的改变

目的神经微丝（ＮＦ）和胶质纤维酸性蛋白（ＧＦＡＰ）分别为神经细胞和胶质细胞特异的中间丝蛋白。着重探讨大鼠脊髓缺血损伤后ＮＦ和ＧＦＡＰ的改变及其可能的病理意义。方法利用ＮＦ和ＧＦＡＰ的单抗，借助于免疫组化、图像分析等手段，对大鼠腹主动脉再灌流缺血模型损伤脊髓神经细胞ＮＦ６８染色强度及胶质细胞数进行定性定量研究。结果伤后１天，ＮＦ的变化不明显；伤后３天，ＮＦ染色明显降低；７天达最低水平，并持续至２１天；２８天基本恢复至伤前。而胶质细胞数与正常相比，伤后１，３，７天无明显变化；至伤后１４天明显增加，并随时间而增加；至２８天达高峰。结论脊髓损伤后ＮＦ的降低与创伤性脑损伤中ＮＦ的变化相似，提示亦具有扩散性轴突损伤的存在。而胶质细胞的增生可能参与脊髓损伤的修复过程。     

胚胎脊髓移植对大鼠损伤脊髓胶质纤维酸性蛋白表达的影响

目的 研究大鼠脊髓损伤后胚胎脊髓（ＦＳＣ）移植是否能够影响胶质纤维酸性蛋白（ＧＦＡＰ）的表达和伤后大鼠后肢功能的恢复情况。方法 将６０只大鼠分为脊髓半脊髓半切洞损伤＿胚胎脊髓移植组（Ａ组）和单纯脊髓半切洞损伤组（Ｂ组）。伤后１,３,,７,１４,２８天,应用行为学和电生理检查观察大鼠功能恢复情况,应用原位杂我和免疫细胞化学方法观察ＧＦＡＰ的表达,并采用计算机图像分析技术,进行定量分析。结果 大鼠脊髓     

脑创伤程度对大鼠伤后胚胎神经干细胞移植的影响

目的 探讨不同程度创伤性脑损伤(traumatic brain injury,TBI)对伤后胚胎神经干细胞(neural stem cells,NSCs)移植的影响. 方法 从孕12～ 14 d胚胎大鼠海马组织中分离NSCs,采用无血清培养法,进行体外培养、扩增和鉴定.大鼠分别于轻型、中型TBI后3d行胚胎NSCs双侧海马区移植;细胞移植14 d后行组织学和TUNEL检测,并对BrdU、NSE、GFAP、GalC、NGF、BDNF蛋白行免疫组化检测. 结果 移植治疗后14 d,轻型TBI组双侧海马区Brdu阳性细胞数明显多于中型TBI组.移植胚胎NSCs脑内分化以GFAP阳性胶质细胞为主.轻、中型TBI后NGF和BDNF蛋白阳性表达增加,其中以轻型TBI组表达最为显著. 结论 轻型和中型TBI对NSCs移植的影响与伤后脑组织局部微环境因素的改变密切相关.     

外周神经损伤对脊髓背角与中枢-外周移行区星形胶质细胞反应的影响

目的 应用免疫组化技术及电镜观察外周神经损伤后脊髓背角中枢--外周移行区星形胶质细胞反应。方法 51只成年wistar大鼠随机分为3组：A组（n=7）正常对照组;B组（n=24）背根压榨伤组;C组（n=20）坐骨神经压榨伤组。损伤组按存活期不同各分为4个亚组,每亚组5例,大鼠分别于伤后3天、2周、1月和2月处死取材。A组、B组3天、2月时相点各取2例用于电镜标本制作。免疫组化染色观察L5脊髓背角及中枢-外周移行区胶质原纤维酸性蛋白（GFAP）的表达,电镜观察中枢-外周移行区星形胶质细胞反应。结果（1）背根和坐骨神经损伤可引起脊髓背角GFAP的显著表达,其中后者的GFAP表达随时间延长而减弱,而前者整个观察期持续高水平表达;（2）坐骨神经损伤不引起中枢-外周移行区GFAP的表达增强,但背根损伤可导致该区域GFAP的显著表达且持续整个观察期;（3）超微结构观察示背根压榨伤后中枢-外周移行区星形胶质细胞突起大量增生,占据了背根传入纤维的径路。结论 外周神经损伤引起的脊髓背角及中枢-外周移行区的反应性胶质化可能是背根再生不能重建脊髓和外周联系的重要原因。     

pSVPoMcat微基因修饰雪旺细胞移植对脊髓损伤的修复作用

为观察pSVPoMcat微基因修饰雪旺细胞（SC）移植对脊髓损伤的修复作用，采用切割法制备脊髓半模断损伤模型（T8平面）。实验动物随机植入pSVPoMcat微基因修饰的SC（A组）、SC组（B组）和损伤对照组（C组），每组40只。术后应用联合行为记分（CBS）和皮层体感诱发电位（CSEP）动态观察大鼠功能恢复情况，应用原位杂交和免疫细胞化学方法观察胶质纤维酸性蛋白（GFAP）的表达。术后3月行MRI观察，并用免疫组化对神经轴突进行神经中丝（NF）染色。结果显示，A组能抑制大学损伤后的GFAP表达；MRI发现，A组大鼠脊髓损伤（SCI）区脊髓信号已基本恢复正常，B组未恢复正常，而C组SCI有软化灶形成。与NF染色发现一致。A、B两组潜伏时波幅呈恢复的趋势，与A组接近正常，与CBS结果一致。提示，pSVPoMcat微基因修饰SC移植能抑制SCI后GFAP的表达，促进神经轴突的生长并对神经功能恢复有明显的促进作用。     

+Gz重复暴露对大鼠脑胶质细胞酸性蛋白表达的影响

目的 探讨＋Gz重复暴露对星形胶质细胞中胶质原纤维酸性蛋白（GFAP）表达的影响。方法 SD大鼠40只，随机分为4组，即对照组（5只）、＋4Gz锻炼组（15只）、＋4Gz锻炼后再暴露于 10Gz组（15只）及单次＋10Gz暴露组（5只）。各组暴露后2d处死取脑，做冰冻切片，免疫组织化学染色观察GFAP的表达情况。结果 4Gz/3min暴露1次、3次和5次后（两次之间间隔24h)，GFAP阳性细胞数由少至多，以细小型为主；单次＋10Gz暴露后，GFAP阳性反应较强，肥大型在梨状皮层和丘脑下部最多，在顶叶皮层和海马较少；＋4Gz分别暴露1次、3次和5次后再暴露于＋10Gz，肥大型在各脑区逐渐减少，在顶叶皮层和海马逐渐消失，代之以细小型。结论 低G值反复暴露后再暴露于高G值对脑的保护作用，与其能减少肥大、肿胀和损伤的GFAP阳性细胞有一定的相关性。