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细胞核

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Results The amaranthine nuclei had a distinct outline and distributed evenly on the slide. The area of nuclei in liquidbased preparation was much smaller and chromatin was more condensed in comparison with those in conventional smear. The nuclei in conventional smears had the largest value of area and the value of AOD, and CV of IOD, the IOD ratio among diploid, tetraploid and octaploid DNA was most close to 2 and 4, and the CV of IOD exceeded 6%.

结果 三种涂片内肝细胞核分布均匀,轮廓清晰,呈紫红色,与传统涂片法相比,液基法内肝细胞核面积明显减小,染色质高度浓缩;相同倍体的肝细胞核在传统涂片中面积最大,平均光密度最低,各倍体间的比值最接近2和4,积分光密度CV值最低(。5)。

Porcine was acted as the experimental animal. It was researched on problem of the origin of obtaining early embryonic cells, methods of separating blastomere cells, best parameter and order of ECNT embryo electric activation and system of ECNT embryo IVC, which was to find a scientific method of fitting ECNT and a procedure of fitting the lab producing embryos largely, optimize the procedure of ECNT and improve the ECNT embryo in vitro.

本实验以猪为实验动物,对猪胚胎细胞核移植供体的来源、卵裂球细胞分离方法、核移植重建胚的电激活最佳参数、核移植受体的激活时序以及核移植重建胚体外培养系统相关问题进行了研究,旨在探索一种适用于猪胚胎细胞核移植的科学方法,提高猪胚胎细胞核移植胚胎的体外发育率,优化胚胎细胞核移植程序。

Morphological and flourescent changes were assessed using confocal microscopy in whole-mount organ of Corti preparations. Results (1) After the animals were exposed to broadband noise at 122 dB SPL in 4 h/day for 2 days, both apoptosis and necrosis appeared in OHCs. The single strand DNA in apoptotic OHCs were observed both in guinea pigs and mice.(2) In normal OHCs, EndoG was distributed outside of nuclei. EndoG translocated from outside to inside of the nuclei in both apoptotic and necrotic OHCs following noise exposure.(3) The MNNG cochlear perfusion and noise exposure both caused the transloctation of AIF from the mitochondria to the nuclei. The translocation of AIF took place in both apoptotic and necrotic OHCs.

结果 (1)暴露于120 dB SPL的白噪声环境中每天4小时,连续2天后引起豚鼠和小鼠耳蜗外毛细胞凋亡时,其细胞核内产生ssDNA,而在正常细胞内没有三ssDNA;(2)在正常情况下,EndoG分布于耳蜗毛细胞的细胞核外,在暴露于上述噪声后发生凋亡和坏死的豚鼠耳蜗外毛细胞中,EndoG从细胞核外转移到细胞核内,细胞核中的EndoG显著增加;(3)豚鼠耳蜗外淋巴灌流烷化剂MNNG后发生耳蜗外毛细胞凋亡和坏死,在凋亡和坏死的耳蜗外毛细胞中,AIF自线粒体转移到细胞核,其变化与噪声损伤引起耳蜗外毛细胞凋亡和坏死时一致。

OBJECTIVE: To investigate a simple easy method for isolation of cell nuclei, and to provide a great quantity of active cell nuclei for studying neuronal nucleus transplantation, cell isolation-fusion and nucleus composition.

目的:探索简单易行的分离细胞核的方法,为神经元细胞核移植、细胞拆合以及细胞核成分的研究提供大量完好有活性的细胞核

In the SVZ, there were clusters of special homotypic nuclei, which were relatively big in size and elliptical in shape, and were named as neuroblast-like nuclei in the present study, for they resembled the migrating neuroblasts in appearance.

在SVZ,可见特异性的同型细胞核聚集现象,此类细胞核体积较大且都呈椭圆体,具有迁移中成神经细胞核特征,故称之为成神经样细胞核

3Rhe distribution and change of proliferating cell nuclear antigen in seminiferous tubule: The positive staining of proliferating cell nuclear antigen was found in slices of all groups, the immunological positive products were located at karyon, usually in brown yellow granules, only a small part of them in diffuse state; the immunological positive cells were mainly the spermospores, besides a small quantity were spermiocyte, supporting the cell and myoid cell nuclear to be stained.

3增殖细胞核抗原在生精小管中的分布及变化:各组切片均可见增殖细胞核抗原阳性染色,免疫阳性产物位于细胞核,通常呈棕黄色颗粒,小部分呈弥散状;免疫阳性细胞主要是精原细胞,另外还有少量初级精母细胞、支持细胞和肌样细胞核着色。

After NMDA stimulation, he concentrations of Ca~2+ and NO in both nuclear and plasma rise observably, especially in the netlike structures; the change of Ca~2+ concentration in nuclear is larger than that in plasma,however the changes of NO concentration in nuclear and plasma are similar. We also observed comparative increases of the concentration of Ca2+ and NO in neurodendrite after NMDA stimulation.

NMDA刺激后神经元细胞核和细胞质内Ca~(2+)和NO都明显升高,尤其在细胞核和细胞质内的网状结构处;细胞核内Ca~(2+)升高幅度大于细胞质,而细胞核内NO的升高幅度却与细胞质相近;另外,树突内的Ca~(2+)和NO升高也较明显。

Fig.1 SHEE cultured on coverslide, the living cells were growing in single layer with rich cytoplasm, the nuclei were uniform in size with a nucleolus ph ×400 Fig.2 SHEE had a nucleus with ellipse shape, large nucleolus and the cytoplasm contained mitochondria and tonofibrilEM ×10 000 Fig.3 SHEE was spherical in shape, with pseudopods attached on petri dish and abundant villi on cell surface SEM ×5 000 Fig.4 Same as in Fig.3, cell attached on petri dish, appeared stellate or polygonal in shape, with abundant pseudopods and cytoplasmic processes. Protrusive nuclear region in central part of the cell had more micro-villi SEM ×5 000 Fig.5 Chromosomes of SHEE cells belonged to diploidy type Giemsa ×1 000 Fig.6 The SHEE cells of stained in dark brown by Ki67 immunohistochemistry were the proliferative cells Immunohistochemistry ×400 Fig.7 In SHEE cell culture, the nucleus stained red or pink by PI was dead cell, the green nucleus was living cell Fluorescent ×400 Fig.8 The cell labeled by TdT was apoptotic cell in which the chromatin of nucleus condensed in block, a pyknotic nucleus in the upper right conner was seen TdT labeled ×400

图1 SHEE培养在盖坡片上,活细胞单层生长,胞浆较丰富,细胞核大小一致,有核仁×400 图2 SHEE培养细胞细胞核椭圆形,核仁较大,胞浆有较丰富的线粒体和张力原纤维EM ×10 000 图3 SHEE细胞呈球状,有伪足贴壁,表面有密集微绒毛SEM ×5 000 图4 同上细胞贴壁,呈星状或多角形,有丰富伪足和胞浆突,核区隆起有较多微绒毛SEM ×5 000 图5 SHEE细胞染色体仍属二倍体Giemsa染色×1 000 图6 SHEE细胞Ki67免疫组织化学染棕黄色为增殖细胞×400 图7 SHEE培养细胞出现死细胞,胞核和胞浆PI染色呈红色或淡红色,蓝色细胞核为活细胞荧光显微镜×400 图8 细胞TdT标记阳性为凋亡细胞,染色质凝集呈块状,右上角有一固缩细胞核TdT标记×400

When stained with TRICT-conjugated phalloidin, both the nuclei and nuclear matrix specimens were found to give off specific, red fluorescent signals which represent the location of F-actin, and the pattern of TRITC signals distribution in the nuclei and nuclear matrix showed similarity with that of FITC signals.

在荧光显微镜下观察到:代表肌动蛋白的特意性荧光分布在细胞核和细胞骨架中,说明肌动蛋白是细胞核和核骨架的固有成分;代表F-肌动蛋白的特异性荧光存在于细胞核和核骨架中,说明细胞核和核骨架含有F-肌动蛋白。

The results show that: 1 supplementation of protein to maturation media improves cumulus expansion in vitro compared to the protein-free control, but cumulus expansion is not necessarily related to oocyte nuclear maturation in pigs, and cumulus expansion is not the criterae for determination of nuclear maturation of pig oocytes, but only the exclusion of the first polar body; 2 exposure of pig COCs to hormone supplements for 23-24 hours improved cumulus expansion but had no significant effect on nuclear maturation compared to that for 46-48 hours; 3 under our research conditions, supplementation of different proteins into different maturation media has different effects on porcine oocyte nuclear maturation, but has no significant effect on subsequent embryonic development after IVF; 4 the nuclear maturation rates of pig oocytes matured in mTCM+pFF and mNCSU+pFF are superior than that in mNCSU+FCS; 5 different maturation media have no effect on pig oocyte cumulus expansion and subsequent embryonic development after IVF.

结果显示:(1)在成熟液中添加蛋白质可以加强卵丘细胞的扩散,但猪卵母细胞的核成熟与其周围的卵丘细胞扩展没有必然的联系,卵丘细胞扩散或成放射状不宜作为猪卵母细胞核成熟的标准,只有排出第一极体才能作为猪卵母细胞核成熟的标志;(2)在猪COCs的46-48小时成熟培养的后23-24小时阶段去除成熟液中的激素不但可以保证卵母细胞的核成熟率,而且可加强卵丘细胞的扩散;(3)在现有实验条件下,在mTCM和mNCSU中添加10%pFF与在mNCSU中添加10%FCS相比可获得较高的猪卵母细胞核成熟率;(4)在不同的成熟液中添加不同的蛋白质对猪卵母细胞核成熟率的影响效果不一样,但对体外受精后的早期胚胎发育影响不明显;(5)成熟液种类对猪卵母细胞的卵丘细胞扩散和体外受精后的早期胚胎发育无显著影响。

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