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In the present study we transfected F-GFP into cultured hippocampal neurons at 5 days in vitro(DIV 5).Then ASICs antagonists were applied to the neuronal medium to inhibit the function of ASICs.Dendritic growth and arborization of cultured hippoeampal neuron were observed in DIV8 and DIV14. We attempted to determine whether and how ASICs regulate dendritic development.

在本研究中,我们在体外培养第五天(days in vitro,DIV5)的原代海马神经元中转染定位在膜上的GFP,随后加入ASICs拮抗剂抑制ASICs的功能,观察DIV8和DIV14这两个时间点海马神经元的树突生长、分支复杂程度,来研究酸敏感离子通道是否会影响海马神经元的树突发育。

The traditional Chinese medicine CN Ⅱ displies different degrees of bacteriostasis to the common pathogenic bacteria.

表明中药肠宁Ⅱ对常见病原菌表现出不同程度的抑菌作用,可以抑制和杀灭体外病原菌的生长发育。

In addition, the above three media could also support the development of 1 cell embryos to hatching blastocyst stage.

实验2的结果表明,在M16培养液中单独添加2.5mmol/L的牛磺酸也可支持体外培养的昆明小鼠1-细胞胚胎发育到囊胚。

The results of experiment shown the hetero-reconstructed embryos could further develop into blastocyst stage in vitro.

实验结果表明,小鼠、兔异种间的核移植重构胚胎可在体外完成到囊胚阶段的早期发育。

Objective To investigate the developmental changes of neonatal bovine retinal horizontal cells in vitro.

目的:研究新生小牛视网膜水平细胞在体外培养中的生长发育规律。

This experiment aimed to systematically investigate and analyze the ethanol activated rates of mouse oocytes, affected factors of oocytes activation, types and development of parthenotes, and the organization and roles of the cytoskeleton during pronuclear formation and migration and early embryo cleavage by ethanol activation of oocytes, embryo culture in vitro and immunofluorescence cyto-chemistry. The results:(1) The ethanol activation of mouse oocytes showed that the activated and developmental rates of oocytes increased significantly with the increase of ethanol concentration and extension of exposure time, but over concentration and exposure time would result in increased fragment rates significantly. 7% ethanol treated oocytes for 7min was the optimum activated condition.

本实验主要利用卵母细胞的乙醇激活、胚胎体外培养和免疫荧光细胞化学方法对小鼠卵母细胞乙醇激活效率、影响因素、孤雌胚类型、发育、原核形成、迁移及早期胚胎卵裂过程中细胞骨架的组装、作用等进行了系统的研究和分析,结果显示:(1)小鼠卵母细胞的乙醇激活结果表明,随着乙醇浓度的升高和作用时间的延长,卵母细胞激活率和发育率都显著提高,但乙醇浓度过高和作用时间过长会导致卵母细胞碎裂率的显著增加,7%乙醇作用卵母细胞7min 为最佳激活条件。

The results of the determinating the activity of the CA in vitro showed that the CA of the water—fed individuals were in activing statics after eclosion. Because of the apperence of the Vg in haemolymph and the deposition of the vitellin in oocytes in the unfed females, it was shown that the CA in such individuals were in activiting statics, too. Again because in the water—fed females which did not lack the feeding action the activity of the CA showed dropping tendency gradually still, it could be said that the activiting of the CA was possibly effected by the nutrition but it was never related to the adult feeding action. In another respect, the brain of the female adult had not significant effect on the activiting CA. The oocytes could depositing vitellin in individuals removed brain tissue on the day of eclosion, which showed that the activting of CA was not concerned with brain.

CA活性的体外测定说明饲水个体的CA同样具有活性,不取食个体血淋巴中Vg的存在以及卵母细胞中卵黄的沉积表明,在不取食的个体中CA同样具有活性,所以CA的活化与成虫期的取食动作无关,与成虫是否取食花蜜也无太多的联系,但是CA活性的维持可能受成虫期营养的影响,如果以卵母细胞发育和CA体积大小作为CA活性的指标,则饲水个体CA的活性在后期受到了影响,说明饲水不能替代取食蜂蜜来维持成虫期CA的活性;成虫期脑对CA的活化无明显作用,羽化当日摘除脑的个体,其卵母细胞仍然可以沉积卵黄,说明CA的活化与脑无关,而是受某种不确定的因子的影响;用抗JH物质—早熟素—Ⅱ以10ug/虫的剂量在颈部点滴处理雌虫,对Vg的发生和卵巢的发育具有一定的迟缓作用,其原因有待于进一步的研究。

Following chemical activation, blastocysts rate of the treated oocytes was similar to untreated oocytes.8 Following fertilization, however, few oocytes inhibited with CHX developed into morulae/blastocysts, due to a high incidence of polyspermy.9 Cortical granule migration occurred during inhibition, but CHX inhibition impaired CG migration, significantly no oocytes inhibited by CHX completed CG migration after maturation.10 CHX inhibition had no effects onα-microtubles and microfilaments of goat oocytes.

抑制24h转为正常培养24h,不影响卵母细胞的成熟和孤雌激活能力,并且CHX抑制后再成熟的卵母细胞经孤雌激活发育到囊胚的比例与对照组卵母细胞相似。8、体外受精后,CHX抑制后再成熟卵母细胞的多精受精现象显著增加,发育到桑椹胚/囊胚的比例显著低于对照组。9、CHX抑制过程中皮质颗粒仍能发生迁移,但是CHX抑制会对皮质颗粒的迁移造成不可恢复的损伤,使再成熟的卵母细胞内皮质颗粒不能完全迁移。10、CHX抑制对山羊卵母细胞α-微管和微丝都没有影响,无论是抑制后处于生发泡期的卵母细胞,还是抑制后再成熟的卵母细胞,微管和微丝的分布都与对照卵母细胞相似。

But embryo got through 2-cell block after H2O2 treatment occurred compensatory growth in later development, there were no differences with the embryo cell number and the cleavage index compared with normal developmental embryo.

但是经H_2O_2处理之后渡过2-细胞期阻滞的胚胎能在以后的发育中发生补偿性生长,其胚细胞数和分裂指数都和体外正常发育的胚胎没有差异。

On the termination date, the cultured explants were all examined by Western blot, HE and transmission electron microscope. Our results showed that after 12-days in culture, the cultivation treated with AS-ODN reduced the synthesis of AMBN and had a deformed dental cusp with thinner enamel matrix. Ultrastructure analyses showed that there was hardly any cisternae of the rough endoplasmic reticulum in the ameloblasts at the tip of the cusp of AS-ODN treatedexplants. However, on average the enamel matrix was thinner compared with that in the control group. Furthermore, the collagen fibers in extracellular matrix were found disorganized. These findings seemed to provide a direct experimental evidence that tended to indicate that the arrested AMBN translation in cultured tooth germs might result in the delay of the tooth development.

经用Western蛋白印迹检测表明,所设计的反义核酸对AMBN InRNA具有良好的封闭效果并成功阻断了牙胚对AMBN的表达;在缺乏AMBN情况下,与对照组相比,实验组牙胚在体外可以继续生长发育至钟状晚期,出现成釉细胞和成牙本质细胞的分化,成釉细胞可以分化成为分泌期型成釉细胞,胞浆中缺少合成蛋白质所必需的粗面内质网和高尔基氏体,缺乏溶酶体,表明对蛋白合成和脚的能力降低;实验组牙胚有牙尖形成和基质分泌,但牙尖形态异常,基质形成减少,牙尖周围基质最厚处为O.6卜m,明显薄于对照组的5.spin,基质中胶原纤维粗细不等,排列稀疏, 3 第四军医大学硕士学位论文未见钙化现象,充分证明了AMBN在牙胚发育中参与釉质基质形成和矿化过程,影响胶原纤维和牙本质基质的合成,促进成釉细胞对蛋白质的合成和釉质基质蛋白降解。

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