βA4在帕金森病脑内表达的研究

目的观察βＡ４在帕金森病脑内的表达，探讨βＡ４表达与神经元变性、缺失的关系。方法用免疫组化的ＡＢＣ法对脑组织切片进行βＡ４表达研究，同时做刚果红染色，观察βΑ４病理形态、阳性程度、脑内分布。结果βＡ４在皮质３、５层中有较强表达。而刚果红染色为阴性。结论βΑ４在脑组织中的表达并非Ａｌｚｈｅｉｍｅｒ病所特有，在ＰＤ病人中亦可见到。βＡ４免疫组化法的阳性结果不同于传统的刚果红染色法。βＡ４与神经元变性、缺失有关     

βA4在皮克病脑中表达的研究

目的　观察β淀粉样蛋白４（β Ａ４） 在皮克病（ Ｐｉｃｋ○ｓ ｄｉｓｅａｓｅ） 脑组织中的表达,探讨β Ａ４ 表达与神经元变性、缺失的关系。方法　用免疫组化 Ａ Ｂ Ｃ法对皮克病脑组织行β Ａ４ 免疫组化染色,观察其病理形态和脑内分布。同时行刚果红染色及 Ｌ Ｈ Ｅ 染色。结果　β Ａ４ 在皮克病脑皮质第３ 、５ 层中均有表达。刚果红染色均为阴性。所有病例可见胶质细胞增生和凋亡。结论　β Ａ４ 在脑组织中的表达并非阿尔茨海默病（ Ａ Ｄ） 所特有,在皮克病中亦存在此现象。β Ａ４ 免疫组化法的阳性结果不同于刚果红染色法。β Ａ４ 表达与神经元变性、缺失有关。     

βA4在弥漫性Lewy体病脑组织表达的研究

观察βＡ４在弥漫性Ｌｅｗｙ体病脑组织中的表达,探讨βＡ４表达与神经元变性,缺的失的关系。方法用免疫组化的ＡＢＣ法进行βＡ４免疫组化染色,观察其病理形态,阳性程度,脑内分布。在免疫组人 色的同时做刚果红染色。     

Alzheimer病与老年脑中β-淀粉样蛋白免疫组化的研究

目的探讨β－淀粉样蛋白（β－ＡＰ）与Ａｌｚｈｅｉｍｅｒ病（ＡＤ）、脑老化之间的关系。方法采用免疫组化方法检查经过临床与病理确诊的３例ＡＤ与６例非痴呆老年对照（ＮＡＤ）脑组织广泛部位中β－ＡＰ的沉积，并且与淀粉样蛋白组化染色的方法如刚果红、爱先蓝（ＳＡＢ）染色相对比。结果发现１ＡＤ脑组织中β－ＡＰ的沉积较ＮＡＤ不仅部位广泛，而且程度严重。２β－ＡＰ的免疫组化染色方法较淀粉样蛋白组化染色方法敏感而可靠。结论β－ＡＰ在脑中的沉积可能与神经元的变性以致智能衰退、ＡＤ的发生有关。β－ＡＰ的免疫组化检测可作为ＡＤ组织学诊断的重要参考。     

Alzheimer病及老年脑神经丝蛋白磷酸化异常

采用定量免疫电镜法、ＰＡＰ法和免疫金银法，对１５例非痴呆患者手术取材的颞皮层进行低温包理免疫胶体金定量分析，并对４例Ａｌｚｈｅｉｍｅｒ病及６例非痴呆患者颞皮层神经丝蛋白进行光镜免疫组化研究。发现Ａｌｚｈｅｉｍｅｒ病患者颞皮层磷酸化神经丝在神经元核周体内有堆积，而老年脑轴突中磷酸化神经丝蛋白较中青年对照组明显减少（Ｐ＜０．００１），但非磷酸化神经丝在老年和青年组间无明显改变（Ｐ＞０．０５）。磷酸化神经丝在Ａｌｚｈｅｉｍｅｒ病和老年脑中的不同分布，可能由两种情况下特异性蛋白激酶／磷酸酶系统不同的功能紊乱所致。     

β淀粉样蛋白与细胞因子S100β表达的相关性研究及其意义

目的　研究大鼠脑内Ａβ４０的沉积与细胞因子Ｓ１００β表达的关系。方法　我们将Ａβ４０定向注射于大鼠的双侧海马,采用免疫组化方法观察了Ｓ１００β的表达。结果　Ｓ１００β免疫组化显示,Ａβ４０处理的大鼠海马区域有细胞因子Ｓ１００β的表达,而在生理盐水对照组的大鼠海马区未出现Ｓ１００β的阳性表达。结论　结果提示Ａβ４０可诱导大鼠海马Ｓ１００β表达。在超过生理性效应的阈值水平之上慢性持续性表达时,Ｓ１００β可能是一种重要的致病因子,具有神经毒性效应,参与Ａｌｚｈｅｉｍｅｒ病的病理活动。Ａβ４０可能是一种重要的病理性刺激因素。     

中风与神经疾病杂志1996年第13卷文题索引

中风与神经疾病杂志１９９６年第１３卷文题索引（按汉语拼音字母顺序排列）ＡＡｌｚｈｅｉｍｅｒ病实验性Ａｌｚｈｅｉｍｅｒ模型鼠脑组织５－羟色胺含量改变的研究（３）：１５０ＢＢｉｎｓｗａｎｇｅｒ病Ｂｉｎｓｗｉｎｇｅｒ病临床研究（４）：２３０Ｃ迟发性神经元坏...     

小檗碱对大鼠脑缺血后MCP-1表达的影响

目的探讨小檗碱处理对大鼠脑缺血后单核细胞趋化蛋白-1（MCP-1）表达的影响及小檗碱对脑缺血的神经保护作用。方法建立大鼠短暂性全脑缺血模型，采用尼氏体亚甲蓝染色观察脑缺血后大鼠脑海马CA1区神经元存活情况；采用免疫荧光染色方法检测脑缺血后大鼠缺血脑组织中MCP-1的表达情况。结果（1）与假手术组比较，脑缺血组大鼠脑海马CA1区神经元明显缺失，而小檗碱处理组大鼠脑海马CA1区神经元存活数明显多于缺血对照组；（2）与假手术组比较，脑缺血组大鼠脑缺血区MCP-1表达显著增多，而小檗碱处理显著降低了大鼠脑缺血区MCP-1的阳性表达。结论脑缺血引起MCP-1表达上调，提示MCP-1可能参与脑缺血损伤。小檗碱可抑制缺血脑组织MCP-1的表达，推测其可能经此途径减轻脑缺血的炎症反应而发挥一定的神经保护作用。     

转化生长因子β-1在全脑缺血再灌注后迟发性神经元坏死时的表达

目的 探讨转化生长因子β-1（TGF-β1）在迟发性神经元坏死时对中枢神经系统的保护与修复作用。方法 采用HE染多功能，尼氏染色，透射电镜，免疫组化及RT-PCR技术。结果 常规光镜及电镜观察均显示再灌注后额叶皮质及海马有不同程度的神经元变性。坏死。免疫组织化学结果及RT-PCR结果均显示皮质与海马区内的TGF-β1在缺血再灌注后呈明显阳性改变。结论 TGF-β1在脑缺血损伤时尤其在发生迟发性神经元坏死时可呈现明显于正常脑组织的表达，因此，TGF-β1不仅在脑缺血早期而且在发生迟发性神经元坏死时，均可能对中枢神经系统产生保护和修复作用。     

大鼠胎脑皮质神经干细胞体外培养及诱导分化的实验研究

目的 从孕龄15d SD胚胎鼠脑皮质中分离并培养神经干细胞（neural stem cells。NSCs），观察其生长、增殖及分化。方法 采用包含碱性成纤维细胞生长因子（bFGF）和表皮细胞生长因子（EGF）的无血清培养及单细胞克隆技术，对胚胎鼠脑皮质神经干细胞进行原代、传代培养及诱导其分化。用Nestin染色鉴定神经干细胞特性，用免疫组化方法（β-Ⅲ-tubulin、GFAP染色）检测神经干细胞分化为神经元及神经胶质细胞状况。结果 从孕龄15dSD胚胎鼠脑皮质中分离的组织，经原代及传代培养均可形成细胞克隆．切具有增殖能力。原代及传代培养细胞呈Nestin（神经上皮干细胞蛋白）表达阳性．诱导分化后的细胞表达神经元细胞、星形胶质细胞的特异性抗原。结论 本实验分离、培养的孕龄15dSD胚胎鼠脑皮质细胞Nestin表达阳性．分化后表达神经元和星形胶质细胞的标记物，是大鼠的神经干细胞，并具有多向分化潜能。     

Congo red and protein aggregation in neurodegenerative diseases

Congo red is a commonly used histological dye for amyloid detection. The specificity of this staining results from Congo red's affinity for binding to fibril proteins enriched in beta-sheet conformation. Unexpectedly, recent investigations indicate that the dye also possesses the capacity to interfere with processes of protein misfolding and aggregation, stabilizing native protein monomers or partially folded intermediates, while reducing concentration of more toxic protein oligomers. Inhibitory effects of Congo red upon amyloid toxicity may also range from blockade of channel formation and interference with glycosaminoglycans binding or immune functions, to the modulation of gene expression. Particularly, Congo red exhibits ameliorative effect in models of neurodegenerative disorders, such as Alzheimer's, Parkinson's, Huntington's and prion diseases. Another interesting application of Congo red analogues is the development of imaging probes. Based on their small molecular size and penetrability through blood-brain barrier, Congo red congeners can be used for both antemortem and in vivo visualization and quantification of brain amyloids. Therefore, understanding mechanisms involved in dye-amyloidal fibril binding and inhibition of aggregation will provide instructive guides for the design of future compounds, potentially useful for monitoring and treating neurodegenerative diseases.     

TAR-DNA binding protein 43 in Pick disease

Pick disease (PiD) is a frontotemporal dementia characterized by frontal and temporal atrophy, neuronal loss, gliosis, ballooned neurons that are positive for alpha-B crystallin and neurofilament, and the presence of tau- and ubiquitin-positive Pick bodies. TAR-DNA binding protein 43 (TDP-43) has been found to be a component of ubiquitinated inclusions in other neurodegenerative diseases, including frontotemporal lobar degeneration with ubiquitinated inclusions and amyotrophic lateral sclerosis. Fifteen cases of PiD were examined using immunohistochemical methods, and 5 cases with both Pick bodies and smaller intracytoplasmic inclusions that showed staining for ubiquitin, tau, and TDP-43 were observed. The presence of TDP-43 inclusions in PiD suggests that TDP-43 accumulation may be an important component of many neurodegenerative diseases, and that its presence in only some cases of PiD may indicate different pathways of disease development.     

Markers for neuronal degeneration in organotypic slice cultures

This protocol describes ways of monitoring spontaneous or induced neuronal degeneration in organotypic brain slice cultures. Hippocampal cultures (4-week-old) are grown in normal serum-free control medium, or exposed to the neurotoxin trimethyltin (TMT) (0.5–100 μM) for 24 h or the excitotoxic glutamate agonist kainic acid (KA) (5–25 μM) for 48 h followed by 24 h or 48 h, respectively, in normal medium. Corticostriatal slice cultures (also 4-week-old) are exposed to KA (6–24 μM) for 48 h and normal medium for control. The resulting neurodegeneration is estimated by (a) propidium iodide (PI) uptake, (b) lactate dehydrogenase (LDH) efflux to the culture medium, (c) ordinary Nissl cell staining, (d) staining by the neurodegenerative marker Fluoro-Jade (FJ), (e) neuronal microtubule degeneration by immunohistochemical staining for microtubule-associated protein 2 (MAP2), and (f) Timm sulphide silver staining for heavy metal alterations. Both hippocampal and corticostriatal slice cultures show a dose- and time-dependent increase in PI uptake and LDH efflux after exposure to TMT and KA. The mean PI uptake and the LDH efflux into the medium correlate well for both types of cultures. Both TMT and KA exposed hippocampal cultures display in vivo patterns of differential neuronal vulnerability as evidenced by PI uptake, FJ staining and MAP2 immunostaining. Corticostriatal slice cultures exposed to a high dose of KA display extensive striatal and cortical degeneration in FJ staining as suggested by a high PI uptake. A change in Timm sulphide silver staining in deep central parts of some control cultures, corresponds to areas with loss of cells in cell staining, loss of MAP2 staining, PI uptake, and FJ staining. We conclude that organotypic brain slice cultures, in combination with appropriate markers in standardized protocols, represent feasible means for studies of excitotoxic and neurotoxic compounds.Themes: Disorders of the nervous systemTopics: Neurotoxicity     

Focal ischemia induces expression of protease-activated receptor1 (PAR1) and PAR3 on microglia and enhances PAR4 labeling in the penumbra

Thrombin significantly influences neurodegenerative processes after ischemia. The current literature suggests that the effects are mediated via protease-activated receptors 1, 3 and 4 (PAR1, 3, 4). Therefore, we investigated with immunohistochemical methods whether focal cerebral ischemia altered the expression of PARs in the rodent brain. For this purpose, we used the model of endothelin-induced occlusion of the middle cerebral artery and the model of transcranial permanent occlusion of the middle cerebral artery in mice. In contrast to the exclusively neuronal staining in the brain parenchyma of na?ve animals, PAR1 and PAR3 occurred in addition on microglial cells in the penumbra after transient and after permanent focal ischemia. Although microglia activation could be detected for several weeks after the insult, PAR1 and PAR3 were traceable on microglia only 12 and 48 h after the insult, but not on day 7 post-ischemia. PAR4 was expressed, both in na?ve and in ischemic animals, exclusively in neuronal cells. However, at the border zone and within the infarct area, enhanced immunohistochemical PAR4 signals were recognized. From our data, we conclude that PAR1 and PAR3 could be involved in thrombin-modulated initiation of post-ischemic inflammation and PAR4 may be associated with neuronal degeneration.     

Ballooned neurons in select neurodegenerative diseases contain phosphorylated neurofilament epitopes

Summary Ballooned neurons are histological features of several neurodegenerative diseases of the central nervous system. We describe the immunocytochemical staining of ballooned neurons in Pick's disease, unclassified dementia, corticonigral degeneration, pigment-spheroid degeneration and Alzheimer's disease. In all of these conditions the ballooned neurons contain phosphorylated epitopes recognized by monoclonal antibodies to neurofilaments, but not epitopes unique to Alzheimer neurofibrillary tangles and Pick bodies. The morphological features and immunohistochemical properties of ballooned neurons in these disorders bear resemblance to swollen neurons produced by neurotoxins that impair axoplasmic transport of neurofilaments. This finding, by analogy, suggests that impaired axoplasmic transport of neurofilaments may be a common mechanism in various dementing neurodegenerative diseases.     

Neuritic sprouting with aberrant expression of the nitric oxide synthase III gene in neurodegenerative diseases

Neuronal loss, synaptic disconnection and neuritic sprouting correlate with dementia in Alzheimer's disease (AD). Nitric oxide (NO) is an important synaptic plasticity molecule generated by nitric oxide synthase (NOS) oxidation of a guanidino nitrogen of L-arginine. Experimentally, the NOS III gene is modulated with neuritic sprouting. In a previous study, NOS III expression was found to be abnormal in cortical neurons, white matter glial cells, and dystrophic neurites in AD and Down syndrome brains. The present study demonstrates the same abnormalities in neuronal and glial NOS III expression with massive proliferation of NOS III-immunoreactive neurites and glial cell processes in other neurodegenerative diseases including: diffuse Lewy body disease, Pick's disease, progressive supranuclear palsy, amyotrophic lateral sclerosis, multiple system atrophy, and Parkinson's disease. However, each disease, including AD, was distinguished by the selective alterations in NOS III expression and sprouting in structures marred by neurodegeneration. Double label immunohistochemical staining studies demonstrated nitrotyrosine and NOS III co-localized in only rare neurons and neuritic sprouts, suggesting that peroxynitrite formation and nitration of growth cone proteins may not be important consequences of NOS III enzyme accumulation. The results suggest that aberrant NOS III expression and NOS III-associated neuritic sprouting in the CNS are major abnormalities common to several important neurodegenerative diseases.     

Immunohistochemical study on the aggregation of ubiquitin in the central nervous system of the transgenic mice expressing a human Cu/Zn SOD mutation

In the present study, we performed immunohistochemical techniques to investigate the changes in ubiquitin expression in the central nervous system of the transgenic mice expressing a human superoxide dismutase 1 mutation (SOD1)G93A. Sections of brains from control mice showed virtually no immunostaining for ubiquitin, whereas sections from SOD1G93A transgenic mice contained numerous granular or linear deposits of ubiquitin. A high density of the processes containing ubiquitin was detected all around the gray matter of the spinal cord of the mutant transgenic mice. Ubiquitin immunoreactivity was also detected in the cerebellum, brainstem and midbrain of transgenic mice. The first demonstration of the distribution of ubiquitin in the whole brains of the transgenic mice may provide clues for understanding the neuronal degeneration mechanism in amyotrophic lateral sclerosis and other neurodegenerative diseases.     

Abnormal cytoskeletal pathology peculiar to corticobasal degeneration is different from that of Alzheimer's disease or progressive supranuclear palsy

An autopsy case of clinically diagnosed “corticobasal degeneration (CBD)” was investigated. In addition to status spongiosus and neuronal achormasia around the central sulcus, cortical pyramidal neurons and thread-like structures were densely stained by Gallyas stain and tau immunohistochemistry, but apparent fibrillary structures like Alzheimer's disease neurofibrillary tangle were absent. Bodian, methenamine-Bodian, Congo red, thioflavin S, or Bielshowsky stains failed to visualize these structures. They were not stained by immunohistochemical stain with anti-ubiquitin antibody. The widespread cytoskeletal pathology, which is distinct from that in Alzheimer's disease or progressive supranuclear palsy, is suggestive of CBD.