成年大鼠纹状体海人藻酸损伤后nestin的表达

目的　 研究成年大鼠纹状体海人藻酸 (kainicacid,KA)损伤后不同时间点nestin的表达并探讨其机制。方法　向成年SD大鼠纹状体内立体定位注射 0 .5 μlKA(1mg/ml) ,分别于手术后第 1、3、7、14天用免疫组织化学法检测纹状体内nestin表达的变化。结果　 正常纹状体内可观察到微弱的淡染nestin样免疫活性阳性细胞 ;大鼠纹状体KA损伤后 1d ,nestin样免疫活性增强 ;第 3天nestin样免疫活性达峰值 ,nestin样免疫活性阳性细胞的胞体肥大 ,突起粗大 ,分支增加 ;此后nestin样免疫活性阳性细胞染色强度减弱 ,其胞体和突起逐渐变小变细。 结论　 成年大鼠纹状体内注射海人藻酸可诱导nestin的大量表达 ,该表达可能与脑损伤后的自身修复相关。     

免疫磁珠法分选胚胎大鼠脑神经干细胞的初步研究

目的：探索应用免疫磁珠间接阳性法分选神经上皮干细胞蛋白（ｎｅｓｔｉｎ）阳性的神经干细胞群的实验条件,为研究神经干细胞的特性与神经干细胞的培养和移植研究创造有利条件。方法：制取胎鼠大脑组织细胞悬液,免疫磁珠法分选胎鼠脑神经干细胞,以流式细胞术检测阳性细胞纯度,以锥虫蓝染色法检测细胞活性。结果：该法分选的ｎｅｓｔｉｎ阳性细胞纯度为９３．０％～９９．７％,其中活性细胞为９２％～９７％。结论：免疫磁珠法分离胎鼠脑神经干细胞群落简便、有效,可以为神经干细胞细胞培养和移植提供细胞来源,也为研究高纯度神经干细胞的特性提供了实验基础。     

IMMUNOHISTOCHEMISTRY OF MEDULLOEPITHELIOMA AND NEURAL TUBE

Immunohistochemistry profiles of medulloepithelioma (from two 2 1-year-old girls who had 2 cerebral medulloepitheliomas and a 35-week postconceptional female infant with congenital posterior fossa tumor) and neural tube are compared. Microscopically, the tumors contained a medulloepitheliomatous component, manifested as tubular epithelial structures lined by pseudostratified columnar epithelium delineated by well-defined basement membranes. In all cases, glial and neuronal differentiation were noted to differing extents. The medulloepitheliomatous components did not exhibit glial fibrillary acidic protein, neuron-specific enolase, or S-100 protein reactivity. Neurofilament, cytokeratin, and epithelial membrane antigen were focally present in one case. Extensive nestin immunopositivity was confined to the basal cell layer of the epithelium, leaving the luminal surface unreactive or slightly reactive. These cells also displayed a reactivity to vimentin and to microtubuleassociated protein type 5 similar to that of cells of the primitive neural tube. The similarity between the immunohistochemical profile of medulloepithelioma and that of neural tube epithelium suggests a possible reexpression of that component of the genome responsible for neural tube growth and differentiation in medulloepithelioma.     

大鼠脑神经干细胞神经巢蛋白表达的流式细胞术检测

检测胎鼠及成年大鼠脑内神经干细胞特异性抗原神经巢蛋白的表达,判明啮齿类哺乳动物脑内神经干细胞存在的可能性及相应的部位。方法：显微手术下采取胎鼠及成年大鼠大脑与皮质下脑组织,间接荧光法标记ｎｅｓｔｉｎ,以流式细胞术检测。结果胎鼠脑内普遍存在ｎｅｓｔｉｎ阳性细胞,成年大鼠脑室旁组织中也存在ｎｅｓｔｉｎ阳性细胞。     

Nestin and vimentin colocalization affects the subcellular location of glucocorticoid receptor in cutaneous melanoma

Aims: Nestin (a neuronal stem cell/progenitor cell marker of central nervous system development), vimentin (which is ubiquitously expressed in mesenchymal cells), and the glucocorticoid receptor (GR, which is involved in the immune response, cell proliferation, and apoptosis) have been shown to interact in embryonic and undifferentiated tissues in modulating cell proliferation. The aim of this study was to analyse nestin, vimentin and GR expression in tumour tissue (melanoma), and their association with clinicopathological variables, to evaluate any effect on tumour progression. Methods and results: Immunohistochemistry, double‐label immunofluorescence and confocal laser scanning microscopy were performed on biopsy specimens of cutaneous melanoma from 81 patients. Fisher’s and Pearson’s tests showed a correlation between nestin, vimentin and subcellular GR location (P = 0.008). Their concomitant expression also correlated with Clark level and thickness (P = 0.02 and P = 0.029, respectively). Kaplan–Meier analysis revealed a poorer outcome for stage III and IV patients with associated expression of nestin, vimentin and cytoplasmic GR in tumour tissue (P = 0.02). Conclusions: These results suggest the presence in melanoma of growth mechanisms involving nestin, vimentin, and GR, similarly to that occurring in embryonic and undifferentiated cells, and may help in understanding tumour biology to provide a molecular basis for clinical therapies.     

巢蛋白在不同发育阶段人胚胎脊髓中的表达变化

目的:研究人胚胎脊髓生长发育过程中巢蛋白(nestin)的变化趋势。方法:采用免疫组织化学方法对不同发育时期(3周～8个月)的人胚胎脊髓中巢蛋白(nestin)的表达及变化进行了研究。结果:Nestin阳性细胞在神经上皮贯穿整个发育过程,5周基板、翼板形成后即可见到阳性物,而边缘层6周出现。中央管、基板、翼板、边缘层内nestin阳性细胞达高峰的时间分别是6周、7周、8周、9周,而表达明显减少的时间分别是8周、9周、10周、11周。随着脊髓的发育成熟,灰、白质内nestin阳性产物逐渐减少,而中央管的阳性细胞数至3月后较为稳定。结论:在脊髓不同部位神经干细胞出现时间和达到高峰的时间均不同,提示在脊髓的不同部位神经干细胞增殖、分化并不同步。     

电针结合减重步行训练对急性脊髓损伤大鼠巢蛋白和神经生长因子表达的影响

摘要 目的：通过检测脊髓损伤后运动功能恢复情况和巢蛋白（nestin）、神经生长因子（NGF）的蛋白表达来探讨电针（EA）结合减重步行训练疗法（BWSTT）干预脊髓损伤（SCI）的作用。 方法：选用健康成年清洁级雄性SD大鼠72只,随机分为假手术对照组（假手术组）、模型对照组(模型组)、电针治疗组(电针组)、电针结合减重步行训练治疗组（电针+训练组）。用美国NYU脊椎冲击损伤仪致大鼠T9—T10段脊髓急性中度损伤模型。BBB运动功能评分对大鼠后肢运动功能的恢复情况进行评估;免疫组织化学技术检测各时间点损伤段脊髓NGF和nestin的表达。 结果：与模型组相比,两治疗组BBB评分显著增加,电针结合减重步行训练组在术后第14天和第28天显著增加,与电针组比较具有显著性差异（P＜0.05）。两治疗组脊髓损伤术后第14天和第28天NGF和Nestin的表达显著增加,但二者没有显著性差异（P＞0.05）。 结论：①电针能够促进脊髓损伤大鼠内源性NGF和nestin的大量表达来促进神经再生。②电针结合减重步行训练对大鼠运动功能的恢复更为有效。     

Clinical value of CD133 and nestin in patients with glioma: a population-based study

Cancer stem cell-related (CSC) markers have been suggested to have promising potentials as novel types of prognostic and predictive markers in gliomas. However no single CSC-related marker is currently used in clinical decisions. The aim of this study was to investigate the prognostic value of CD133 and nestin separately and in combination using a novel quantitative approach in a well-characterized population-based cohort of glioma patients. The expression of CD133 and nestin was measured by systematic random sampling in stained paraffin sections from 239 glioma patients diagnosed between 2005 and 2009. We found that the expression of CD133 did not correlate with WHO grade, and there was no association with overall survival (OS). The level of nestin correlated positively with WHO grade. In patients with WHO grade II tumors, a high level of nestin was associated with short progression-free survival (PFS) in multivariate analysis. High levels of co-localization were associated with poor PFS in patients with WHO grade II tumors, but not with OS. We conclude that CD133 was not an independent prognostic factor, but a high level of nestin was associated with poor PFS in patients with WHO grade II tumors. The combination of double-immunofluorescence and automated analysis seems to be a feasible and reproducible approach for investigation of the prognostic potential of biomarkers.