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281.
Immunoelectron microscopic studies on centromere-kinetochore complexes detached from chromosomes 总被引:1,自引:0,他引:1
The centromere-kinetochore complexes of Chinese hamster ovary (CHO) cells were detached and separated from the condensed chromatin by treatment with hydroxyurea and caffeine. By labelling the complex for immunoelectron microscopy (immuno-EM) with a mixture of antibodies against centromere proteins (anti-CENP-A,-B, -C) in some cells, we could demonstrate complete detachment of the complexes. No remnants were left at the bulk of condensed chromatin in these cells. In some mitotic cells complex and chromatin were found side by side. It could be shown that the fine structure of the separated material of the complex differs significantly from that of the rest of chromatin. The complex consists of proteins and DNA. This leads us to suppose that the organization of chromatin in the centromere-kinetochore complex is different. 相似文献
282.
L H Anderson T L McDonald G M Thiele L W Klassen 《Journal of clinical laboratory analysis》1992,6(3):113-118
Different assay systems have been used to quantitate lymphokine-induced natural cytotoxic activity as a measure of immune status. This study compares the effects of inducing cytotoxicity in a bulk culture system, where effector cells are transferred to a micro culture well for assay, to a micro culture system where the effector cells are not transferred. The effector/target ratio for both the bulk and micro culture systems was calculated using the number of viable effector cells present at the time of target cell addition. After overnight incubation with interleukin-2 (IL-2), the lytic activity of murine spleen cells to targets using a micro culture system was increased two-fold over the bulk culture method. This increase was amplified further after 5 days of activation with IL-2, in that the micro culture system resulted in a four-fold increase in cytotoxic activity. The loss of some adherent cells in the bulk culture system did not explain the overall decrease in recovered cytotoxicity. The difference appeared to be related to cell loss during centrifugation. Therefore, the E/T ratios are different in the two systems if not corrected for the number of viable cells. 相似文献
283.
白细胞介素-10诱导的大鼠树突状细胞体外免疫功能的研究 总被引:19,自引:7,他引:12
目的 研究白细胞介素 10 (IL 10 )诱导的大鼠未成熟树突状细胞 (imDCs)体外诱导免疫耐受的可行性。方法 在经典诱导方案的基础上 ,应用IL 10 ( 10 μg/L)抑制大鼠骨髓来源DCs的成熟 (IL 10组 ,10例 ) ,并设对照组 (IL 4组 ,10例 )。培养期间观察DCs形态 ,检测DCs表型、摄取抗原能力、体外免疫功能及培养上清细胞因子水平。结果 与IL 4组比较 ,IL 10组DCs细胞表面CD80 、CD86及OX6低度表达 ( 2 5 .3 %、42 .4%、3 2 .3 % ) ,吞噬能力较强 ( 81.9) ,刺激同种异体淋巴细胞增殖能力下降 ,该淋巴细胞具有抗原特异性低反应性 ;培养上清中IL 12水平 ( 4 0 6.5pg/L)及初次MLR培养上清IL 2水平 ( 2 45 .4ng/L)均较低 ,差异有非常显著性 (P <0 .0 1)。 结论 IL 10作用的大鼠imDCs具有诱导免疫耐受的应用价值。 相似文献
284.
285.
286.
病理性损伤因素对肾小管上皮细胞表型转化的影响 总被引:4,自引:0,他引:4
目的 :观察低血清、高糖、高蛋白刺激下肾小管上皮细胞向间叶性细胞的表型转化 ,探讨损伤的肾小管上皮细胞在肾小管间质纤维化中的作用。方法 :以体外培养的人近端肾小管上皮细胞为对象 ,分别用含低血清(0 .2mol·L-1小牛血清 )、高葡萄糖 (4.5 g·L-1)、高白蛋白 (15 g·L-1)的培养液培养 7d ,透射电镜检测细胞形态变化 ;免疫组化和Westernblot检测细胞表型改变 ,包括CK、Vimentin、α SMA、Ⅰ和Ⅲ型胶原 ,原位杂交检测Ⅰ型胶原mRNA表达 ,同时检测细胞TGFβ1蛋白表达。 结果 :低血清、高糖、高蛋白直接刺激下 ,肾小管上皮细胞出现明显形态学改变 ,包括细胞变为长梭形 ;透射电镜下细胞内线粒体明显减少 ,粗面内质网明显增加 ,并出现actin样微丝。免疫组化染色和Westernblot显示CK表达明显减弱 ,Vimentin、Ⅰ、Ⅲ型胶原、α SMA和TGFβ1表达增强。原位杂交显示Ⅰ型胶原mRNA表达增强。结论 :低血清、高糖、高蛋白可直接引起人近端肾小管上皮细胞TGFβ1表达增加 ,并发生向间叶性细胞的表型转化 相似文献
287.
人骨髓间充质干细胞体外分化为雪旺样细胞的方法 总被引:4,自引:0,他引:4
目的 :建立人骨髓间充质干细胞体外向雪旺样细胞定向诱导分化的方法。方法 :取健康人骨髓血标本 ,利用percoll(密度为 1.0 73 g·ml-1)分离骨髓单个核细胞 ,在DMEM LG + 10 % (体积分数 )FBS中培养 ,通过流式细胞术对培养细胞进行表型测定 ,对第 2、3、5、8代间充质干细胞进行定向诱导分化 ,用单抗S 10 0、神经胶质纤维酸性蛋白 (glialfibrillargacidicprotein ,GFAP) ,通过免疫组化方法对诱导的细胞进行鉴定。 结果 :经 percoll分离的间充质干细胞可传 16代 ,细胞数增加了大约 6× 10 7倍。流式细胞术结果显示 :percoll分离出来的未经培养的细胞 ,其CD2 9+CD4 4 +CD3 4 -CD4 5 -细胞数为 3 2 .4 7%± 3 .4 9% ,而培养后贴壁细胞中CD2 9+CD4 4 +CD3 4 -CD4 5 -细胞数为 94 .3 8%± 1.5 0 %。诱导后免疫组化染色结果显示 :由 β ME +bFGF预诱导、β ME +bFGF诱导的S 10 0阳性细胞最多 ,可达 90 %± 4 % ,GFAP阳性细胞为 2 1%± 5 %。结论 :人骨髓中间充质干细胞能定向诱导分化成雪旺样细胞。 相似文献
288.
H. B. Jones C. R. Jenkins A. L. Bowdler M. G. Simpson E. A. Lock 《Acta neuropathologica》1997,93(3):241-251
The objectives of the studies described were to assess the ultrastructural neuropathology, blood-brain barrier (BBB) integrity
and calcium status of the cerebellum of rats following a single dose of 750 mg · kg–1
l-2-chloropropionic acid (l-2-CPA). The first indications of intoxication appeared at 36 h when condensation of many granule cells associated with Purkinje
cell degeneration and marked astroglial swelling were observed. Some electron-lucent granule cells were also noted lying amongst
these condensed forms. Condensed granule cells had swollen, electron-lucent mitochondria, dilated Golgi apparatus and nuclear
crenation. Occasionally, areas of granule cell necrosis were also present at this time. Granule cell condensation probably
represents a preliminary and irreversible stage in an excitotoxic process that leads to necrosis. At 48 and 72 h, most granule
cells were necrotic, and occasionally, extravasation of both erythrocytes and leucocytes into the expanded extravascular space
was observed. Evaluation of the BBB by ultrastructural cytochemical visualisation of horseradish peroxidase injected i.v.
2 min before killing by perfusion fixation showed substantial leakage. At 36 h post-dose, ultrastructural calcium localisation
using oxalate/pyroantimonate precipitation demonstrated a substantial increase in calcium pyroantimonate precipitate in mitochondria
and other membranous cytoplasmic organelles (especially the Golgi apparatus) in condensed granule cells, but with little in
their nuclei. However, their immediate neighbours (of ostensibly normal ultrastructural appearances) contained greater amounts
of intranuclear precipitate. Swollen astroglial cells (especially the Bergmann glia) contained considerable quantities of
precipitate. A possible excitotoxic mechanism via l-2-CPA-induced NMDA receptor agonism leading to overwhelming calcium influx and disruption of cellular calcium homeostasis
is proposed.
Received: 8 May 1996 / Revised: 1 September 1996 / Accepted: 11 September 1996 相似文献
289.
(沈张悦)(邵静芳)(王晓林)(朱慧芬)ExperimentalStudyonAnti-tumorEffectofSplenocytesInducedbyAnti-CD3McAb,PHAandIL-2¥SHENGuan-xin,WANGXiao-li... 相似文献
290.
MRI of the brain in chronic carbon monoxide poisoning 总被引:3,自引:0,他引:3
We examined 13 patients with chronic carbon monoxide (CO) poisoning by magnetic resonance imaging (MRI); all of them had been in an explosion in a coal mine 25 years previously. Symmetrical globus pallidus lesions were observed in 12, as was degeneration of the white matter, with focal cortical atrophy. The temporal parietal and occipital lobes were usually affected, the parietooccipital region being the most frequently and extensively damaged. Of the 12 patients with white matter degeneration 7 had definitely asymmetrical cortical and subcortical lesions. There were 6 patients with dilated temporal horns, probably due to atrophy of the hippocampal gyri. A history of CO inhalation and an awareness of the typical distributions of lesions are important for recognition of the effects of CO poisoning, especially when patients are in the chronic stage. 相似文献