胚胎

To isolate, cultivate and identify the human embryonic fibroblasts derived from the gonadal ridges and dorsal mesenteries of human embryos, and to detect the expression by hEFs of cytokines crucial for the growth of human embryonic germ cells in vitro.

目的：从人胚胎生殖腺嵴和肠背系膜中分离、培养及鉴定人胚胎成纤维细胞，检测hEFs表达人胚胎生殖细胞体外生长所需的重要细胞因子。

Our study includes four aspects. In the first aspect we study several important conditions of porcine oocytes maturation in vitro and oocytes cleavage after parthenogenetic activation and found mNCSU-23+15IU/mlPMSG+20IU/mlHCG+15% PFF+0.57mMcysteine is a good culture condition .When the Cocs are cultured in it ,the maturation rate and oocytes cleavage rate are higher than those of foreign covered. Our result are (86.7±3.35)% and (86.3±4.16)% and the highest report of foreign is(85.7±4.1)%.In the second aspect we study the effect of different chemical activations on development of porcine parthennogenetic embryo and found two best activation method. The first one is that putting the maturation MII oocytes in the 20μmol/L ionomycin for 30 minutes and then putting them in the NCSU-23 condition containing 5μg/mICB and 5mM/L6-DMAP for 3.5 hours, the oocytes cleavage rate and morulae/blastocysts development rate are (76.7±7.6)% and (37.1±6.4)%.The second one is that putting the maturation MII oocytes in the 200μM/L Thimerosal for 20 minutes and then putting them in the NCSU-23 condition containing 8mM DTT for 30 minutes

本研究分为4个部分，第一部分对影响猪卵母细胞体外成熟和孤雌激活后胚胎分裂的几个重要条件进行了比较研究，确立了一种较好的培养方法：与颗粒细胞共培养，找到了一种适合猪卵母细胞体外成熟的培养基：mNCSU-23+15IU/mlPMSG+20IU/mlHCG+15%PFF+0.57mM半胱氨酸，成熟率和分裂率分别为(86.7±3.35)%和(86.3±4.16)%，国外报道的最高成熟率为(85.7±4.1)%；第二部分对猪卵母细胞孤雌激活的化学方法进行了研究，确立了化学激活猪卵母细胞的两种最佳方法：1将成熟的去卵丘颗粒细胞的MII期卵母细胞用20μmol/Lionomycin作用30min，再将卵母细胞培养于含5μg/mlCB和5mM/L 6-DMAP(6-二甲基氨基嘌呤)的NCSU-23培养液中，卵裂率和桑囊胚发育率达到(76.7±7.6)%和(37.1±6.4)%2将成熟的去卵丘颗粒细胞的MII期卵母细胞在200μM/L的Thimerosal中处理20min，再与8mM的DTT共孵育30min，卵裂率和桑/囊胚形成率为(81.0±2.8)%和(39.6±2.7)%；第三部分对孤雌激活胚胎的培养条件进行了研究，确立了一种最佳的胚胎培养条件：在SOF简单培养基中添加颗粒细胞进行前3天的培养，然后转入添加胎牛血清的NCSU—23培养基并和输卵管上皮细胞进行后期的培养，其桑椹胚和囊胚的发育率为(59.5±3.2)%；第四部分研究了IGF-I

The experiment has built a method to sexing the hybridization of chicken and quail in early stage of embryo development.

推测杂交种胚胎死亡的时间集中在69-72h，这一时间段胚胎的死亡，是导致整个孵化过程胚胎死亡率升高的主要原因。

In rat esophagus, the positive varicosities and fibres could be observed in the circular muscle layer and the muscularis mucosae layer at 21st day before birth. With the development of rat digestive tract, neurokinin A-immunoreactivity positive nerves could be observed gradually in the epithelium, submucosa, longitudinal muscle layer, myenteric plexus and submucosal plexes, while mature nerve fibres could be seen at one month after birth. 2. In rat stomach, the positive reaction of NKA initially happened in the myenteric plexus at the 14th day of embryo, and then appeared on circular muscles, longitudinal muscles, submucosal, muscularis mucosae, lamina propria and epithelium. 30 days after birth, expression of NKA is same as seen in adaulthood. 3. In rat small intestinal, the NKA-IR could first be found in the myenteric plexus of the duodenum, jejunum and ileum at 14th , 15th and 17th day before birth respectively. Then the NKA-IR occurred in the longitudinal muscle layer, circular muscle layer, intestinal villus, intestinal gland, muscularis mucosae, submucosa, submucosal plexes, mucosa plexus and deep muscular plexus.

结果 1、在食管，于胚胎21天的粘膜肌层和环肌层内观察到神经激肽A免疫阳性膨体纤维，出生后，随幼鼠的生长发育，相继在上皮，粘膜下层，纵肌层、肌间丛、粘膜下丛有NKA-IR表达，30天时已和成年鼠相似；2、在胃，首先于胚胎14天的肌间丛出现NKA-IR的表达，随发育相继在环肌、纵肌，粘膜下层、粘膜肌、固有膜及上皮内出现NKA-IR的表达，30天时具备成年鼠的分布特征；3、在小肠，分别于胚胎14、15、17天的十二指肠、空肠和回肠的肌间丛处出现NKA-IR的表达，随发育相继出现在纵肌、环肌、绒毛、小肠腺周、粘膜肌、粘膜下层、粘膜下丛、粘膜丛、深肌丛。

Methods Mouse endometrial epithelial cells were isolated and cultured,then it was cocultured with mouse embryos fertilized in vitro.

对小鼠子宫内膜上皮细胞进行分离、培养鉴定，然后与体外受精的小鼠胚胎进行共培养，观察共培养对小鼠胚胎卵裂率、孵化率以及囊胚期胚胎总细胞数的影响。

The results showed that there are two main fastigiums in death curve during the hatching penod, one is in 4-6 day and another is in 17-19 day. The rest embryo ages were divided into three phases by the two pinnacles and the difference of died embryo among three phases is not significant. The results in this paper is not incompletely consistent with commonly experience value.

结果表明，整个孵化期间，胚胎死亡主要有2个高峰，第1个高峰在孵化前期的4~6日龄，第2高峰期在孵化后期的17~19日龄，这2个胚胎死亡高峰期把剩下的各胚龄分为3个阶段：1~3日龄、7~16日龄、20~21日龄，这3个阶段的胚胎死亡皆差异不显著，本研究结果与一般经验值不完全一致。

ES cells were maintained on gelatin-coated dishes in Iscove's modified Dulbecco medium containing 0.1 mmol/L of nonessential amino acids, 2 mmol/L of L-glutamine, 0.1 mmol/L of monothioglycerol, 15% fetal bovine serum, and 1000 U/mL leukemia inhibitory factor. Embryonic bodies were firstly formed by ES cells with hang drip method in IMDM without LIF. Six-day-old EBs were trypsinized and the cells were resuspended at a concentration of （3-4）×109 L-1 cells per 1 mL of a 1:1 mixture of culture medium and Matrigel?

实验方法：将鼠胚胎干细胞株接种于被覆明胶的培养瓶内，加入含0.1 mmol/L非必需氨基酸、2 mmol/L L-谷酰胺、0.1 mmol/L二羟基丙硫醇、15%胎牛血清、1 000 U/mL白血病抑制因子的IMDM培养液。3 d后采用&悬滴法&使细胞在不含白血病抑制因子的上述IMDM培养液中形成胚胎体，6 d后胰酶消化得到胚胎干细胞，浓度调整为（3~4）×109 L-1，与Matrigel？

The sixth, there were no significantly differences between colchicine groupsand vinblastine groups on the metaphases, in the experiment of prepare mouse4-cell embryo stage single blastomere chromosome specimen by using optimumparameters; and there were significantly differences between colchicine groupsand vinblastine groups on the metaphases (P＜0.05), in the experiment of preparemouse 8-cell embryo stage single blastomere chromosome specimen by usingoptimum parameters.The end, the preparation quality of mouse 4-and 8-cell embryo stage singleblastomere specimen were 2.5, 2.7, 2.4, 1.9, respectively.

低渗液选用0.45％氯化钠时，低渗效果优于选用0.5％柠檬酸钠和0.35％氯化钾的。6、在利用最佳参数制备小鼠4-细胞期胚胎单卵裂球染色体标本时，经统计分析，两种阻断剂的作用效果差异不显著；而在制备小鼠8-细胞期胚胎单卵裂球染色体时阻断剂选用长春花碱的效果优于秋水仙素的(P＜0.05)。7、秋水仙素组和长春花碱组小鼠4、8-细胞胚胎单卵裂球染色体标本制备质量分别为：2.5、2.7、2.4、1.9。

The cleavage rate of 2-cell stage was 50%, but few embryos developed into morulas and blastocysts (2%). More than 130 embryos at 2-4-cell stage were transferred into oviduct of 13 synchronized foster rabbits and 2 mice. No fetus or implantation sites were observed in both recipients.

其2—细胞胚胎的分裂率可达50％，但发育至桑椹胚和囊胚阶段的胚胎数目大大减少，只有2％的囊胚发育率。13只受体兔共移植重构胚胎121枚，结果未发现产仔或死胎，仅看到有的兔子宫有於血状。

The endurance abilities of the embryos of sea perch, flounder and turbot at different developing stages to vitrification solutions were studied. The result showed that sea perch at tail-bud stage, heart-beating stage and pre-hatching stage had more endurance abilities to VSD2, flounder at 4-5 somites, 16-20 somites and tail-bud stage had more endurance abilities to PM3, and turbot at 4-5 somites, 16-20 somites and tail-bud stage had more endurance abilities to PMP1. The feasible equilibration time of three marine fish embryos in vitrification solutions was determined, which guided the dealing process before freezing.

对花鲈、牙鲆、大菱鲆三种鱼类不同时期胚胎分别在玻璃化液中的适应性进行了研究，发现花鲈尾芽胚、心跳胚和出膜前期胚对VSD2的耐受能力较强；牙鲆4-5以肌节胚、16-20对肌节胚、尾芽胚对PM3的耐受能力较强，大菱鲆4—5对肌节胚、16—20对肌节胚、尾芽胚对PMP1的耐受能力较强，较适合于进行玻璃化液处理和冷冻保存；并且确定了三种鱼类各时期的胚胎在玻璃化液中的适宜平衡时间，为胚胎在玻璃化液中处理和冷冻提供了一定的参数。

According to the clear water experiment, aeration performance of the new equipment is good with high total oxygen transfer coefficient and oxygen utilization ratio.

曝气设备的动力效率在叶轮转速为120rpm～150rpm时取得最大值，此时氧利用率和充氧能力也具有较高值。

The environmental stability of that world - including its crushing pressures and icy darkness - means that some of its most famous inhabitants have survived for eons as evolutionary throwbacks, their bodies undergoing little change.

稳定的海底环境─包括能把人压扁的压力和冰冷的黑暗─意谓海底某些最知名的栖居生物已以演化返祖的样态活了万世，形体几无变化。