体外发育的

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。

Early embryos dissected from host larvae hemocoel grow bigger in size and proliferate by the invagination of extroembryonic membrane, but they can not develop into the individual stage.

从寄生初期寄主幼虫体内得到的初级胚胎可以在体外培养条件下长大并持续通过胚外膜凹陷进行增殖，但无法进入个体发育阶段，胚胎最后黑化死亡。

The result s showed that the outgrowth of neurites as well as neuroglia cells from SCG were significantly increased when SCG were incubated with non-lymphoid conditioned medium compared with that of control medium.

结果显示，与对照培养液相比，条件培养液能够显著地促进体外培养中颈上节交感神经生长锥的发育，且这一作用在不同程度上可被抗NGF和NT-3抗体特异性阻断。

Objective To compare the proliferation and differentiation in vitro of neural stem cells isolated from different periods of ontogeny .

目的比较个体发育不同阶段所得神经干细胞的体外增殖和分化情况。

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。

According to the characteristics of totipotency , differentiation out of body and the steady transmissibility of the ES cells, we can see wide application prospects of the ES cells which could be used to build the model of early diffierentiation out of body and the model of transgenic animal ,to restore apparatus and tissue and transplant therapy, to produce clone animal and research .

人们利用ES细胞所具有的全能性、体外分化以及稳定的遗传性能等特点，展示了ES细胞在建立哺乳动物的早期胚胎体外分化模型、转基因动物模型、器官和组织的修复与移植治疗、克隆动物的生产、发育生物学的研究等方面广阔的应用前景。

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。

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。

In the present study, we collected cumulus cells oocyte complex from ovaries of two different strain mice. The cumulusenclosed oocytes were cultured for 6 h in MEM supplemented with growth factor and FSH. The meiotic maturation of these oocytes has progressed to pro-metaphse Ⅰ stage and the condensed chromosomes are visible under DIC microscope, metaphase Ⅰ spindle even can be detected under Polscope. The metaphase Ⅰ spindles of oocytes were exchanged under such microscopes. After electric stimuli, 91. 6% and 91. 6% karyoplasts-cytoplasm pairs were fused respectively. The resulting oocytes were cultured further in MEM and over 80% of oocytes released the first polar body. 79% and 77% of oocytes formed two pronuclei after in vitro fertilization and the embryos were cultured in KSOM supplemented with amino acids. Over 60% of embryos developed to blastocyst stage.

在本研究中我们在取得两种不同品系小鼠的卵丘卵母细胞复合体后，先将卵丘卵母细胞复合体置于含有多种生长因子和激素的MEM培养液中培养6小时，此时卵母细胞已进入第一次减数分裂的前中期，并且在DIC倒置显微镜下可以看到浓缩的染色体，用Polscope可以发现明显的纺锤体，借助这种显微镜通过显微操作将两种不同品系小鼠来源的卵母细胞的MI纺锤体进行互换，经过三次直流电脉冲作用后，分别有91.6％的胞质—MI核质体对融合，经过进一步的培养后，超过80％的重组卵母细胞排出第一极体，体外受精后分别有79％和77％的重组卵形成双原核，受精后的胚胎在KSOM胚胎培养液中体外培养4天后，超过60％的胚胎发育至囊胚。

But we don't care about Battlegrounds.

但我们并不在乎沙场中的显露。

Ah! don't mention it, the butcher's shop is a horror.

啊！不用提了。提到肉，真是糟透了。