A series of experiments were conducted to study the major procedures in nuclear transplantation such as oocyte enucleation and activation, electrofusion and developnent of the nuclear transplant embryos in the mouse, ...A series of experiments were conducted to study the major procedures in nuclear transplantation such as oocyte enucleation and activation, electrofusion and developnent of the nuclear transplant embryos in the mouse, rabbits and sheep. The important results are as follows:11. In the mouse, only 35% of the oocytes collected 15～16 h after hCG had a notable first polar body (FPb) and those without FPb were enucleated by removing cytoplasm from the PVS-wider side and the enucleation rate was similar to that in the oocytes with FPb, and the enucleation rate of removing 1/3 cytoplasm was remarkably higher than that of removing 1/4 cytoplasm. 2. Among the three fusion media tested, mannitol and sucrose solutions produced better results than M2 in electrofusion of mouse 2-cell embryos. Under favorable pulse conditions, the osmotic pressure of fusion medium had no motable effect on electrofusion, but as the conditions became so unfavorable that some embryos began to lyse, the fusion rates in hypertonie mannitol solution were significantly higher than those in isotonic or hypotonic solutions. A wide range of pulse strengths (0.31～2.04 by/ cm) and durations(10～1280 μs) were used and 100% of fusion were obtained in many cases. Optimal pulse durations were plotted for field strengths to obtain high fusion rates (96%～ 100%) in mouse2-cell embryos. 3. With one pulse of 0.45 by / cm, satisfactory results of mouse oocyte activation were obtained only when the duration increased to 160 μs or longer. The activation rate increased as the oocytes got older. Some of the oocytes ar. rested at metaphase Ⅲ after electrical stimulation and their proportion to the number of oocytes not activated increased with egg age. 4. 10% and 31% of the nuclear transplant embryos developed to morula or blastocyst stage in sheep and rabbits, respectively, with Chinese-made hormones and chemicals.展开更多
目的:研究在IVF-ET的新鲜取卵周期,增加1枚非优质胚胎的移植对妊娠结局的影响。方法:回顾分析2015年1月至2018年12月在郑州大学第二附属医院生殖中心接受IVF-ET助孕的患者的资料。按移植胚胎数分为3组:移植1枚优质胚胎97个周期(组1),移...目的:研究在IVF-ET的新鲜取卵周期,增加1枚非优质胚胎的移植对妊娠结局的影响。方法:回顾分析2015年1月至2018年12月在郑州大学第二附属医院生殖中心接受IVF-ET助孕的患者的资料。按移植胚胎数分为3组:移植1枚优质胚胎97个周期(组1),移植2枚优质胚胎1810个周期(组2),移植1枚优质胚胎和1枚非优质胚胎279个周期(组3)。比较3组的临床资料和妊娠结局。结果:组2的获卵数、优胚数、HCG日E 2值、P值、临床妊娠率和多胎妊娠率均高于组1,差异均有统计学意义( P <0.05)。组3的着床率明显低于组1,多胎妊娠率明显高于组1,差异有统计学意义( P <0.05),临床妊娠率与组1无显著差异。Logistics回归显示,年龄是妊娠结局的危险因素,HCG日内膜厚度是保护因素。结论:增加移植优质胚胎数可提高临床妊娠率,增加1枚非优质胚胎的移植,不能提高临床妊娠率,由于多胎妊娠率明显增高反而增加了妊娠风险。展开更多
通过远缘杂交及回交将白菜的抗病基因导入到芥菜中,创制获得优质的抗根肿病长柄芥(Brassica juncea var.longepetiolata Yang et Chen)育种材料。采用人工接种鉴定和分子标记鉴定相结合法,对10份抗根肿病白菜材料进行抗性分析,筛选出1...通过远缘杂交及回交将白菜的抗病基因导入到芥菜中,创制获得优质的抗根肿病长柄芥(Brassica juncea var.longepetiolata Yang et Chen)育种材料。采用人工接种鉴定和分子标记鉴定相结合法,对10份抗根肿病白菜材料进行抗性分析,筛选出1份可作为长柄芥抗根肿病育种材料创制的抗性资源CCR21002。利用CCR21002分别作为父本、母本提供抗根肿病基因,与长柄芥进行远缘杂交,结合胚挽救技术获得正反杂交种F1,选取植株形态偏向芥菜的F1与长柄芥回交获得BC_(1),对BC_(1)植株进行形态学、细胞学分析及抗病性鉴定。结果表明,回交一代比杂种一代更难获得,胚挽救成活的13株BC_(1)中,仅筛选出1株基因型为AABB且抗根肿病的植株,经分子标记鉴定所含抗病基因为CRb,在植株形态上保留有长柄芥的大部分性状。证明经过与抗根肿病白菜远缘杂交及回交,获得对4号生理小种表现抗性的长柄芥新种质1份,可用于继续回交转育。展开更多
文摘A series of experiments were conducted to study the major procedures in nuclear transplantation such as oocyte enucleation and activation, electrofusion and developnent of the nuclear transplant embryos in the mouse, rabbits and sheep. The important results are as follows:11. In the mouse, only 35% of the oocytes collected 15～16 h after hCG had a notable first polar body (FPb) and those without FPb were enucleated by removing cytoplasm from the PVS-wider side and the enucleation rate was similar to that in the oocytes with FPb, and the enucleation rate of removing 1/3 cytoplasm was remarkably higher than that of removing 1/4 cytoplasm. 2. Among the three fusion media tested, mannitol and sucrose solutions produced better results than M2 in electrofusion of mouse 2-cell embryos. Under favorable pulse conditions, the osmotic pressure of fusion medium had no motable effect on electrofusion, but as the conditions became so unfavorable that some embryos began to lyse, the fusion rates in hypertonie mannitol solution were significantly higher than those in isotonic or hypotonic solutions. A wide range of pulse strengths (0.31～2.04 by/ cm) and durations(10～1280 μs) were used and 100% of fusion were obtained in many cases. Optimal pulse durations were plotted for field strengths to obtain high fusion rates (96%～ 100%) in mouse2-cell embryos. 3. With one pulse of 0.45 by / cm, satisfactory results of mouse oocyte activation were obtained only when the duration increased to 160 μs or longer. The activation rate increased as the oocytes got older. Some of the oocytes ar. rested at metaphase Ⅲ after electrical stimulation and their proportion to the number of oocytes not activated increased with egg age. 4. 10% and 31% of the nuclear transplant embryos developed to morula or blastocyst stage in sheep and rabbits, respectively, with Chinese-made hormones and chemicals.
文摘目的:研究在IVF-ET的新鲜取卵周期,增加1枚非优质胚胎的移植对妊娠结局的影响。方法:回顾分析2015年1月至2018年12月在郑州大学第二附属医院生殖中心接受IVF-ET助孕的患者的资料。按移植胚胎数分为3组:移植1枚优质胚胎97个周期(组1),移植2枚优质胚胎1810个周期(组2),移植1枚优质胚胎和1枚非优质胚胎279个周期(组3)。比较3组的临床资料和妊娠结局。结果:组2的获卵数、优胚数、HCG日E 2值、P值、临床妊娠率和多胎妊娠率均高于组1,差异均有统计学意义( P <0.05)。组3的着床率明显低于组1,多胎妊娠率明显高于组1,差异有统计学意义( P <0.05),临床妊娠率与组1无显著差异。Logistics回归显示,年龄是妊娠结局的危险因素,HCG日内膜厚度是保护因素。结论:增加移植优质胚胎数可提高临床妊娠率,增加1枚非优质胚胎的移植,不能提高临床妊娠率,由于多胎妊娠率明显增高反而增加了妊娠风险。
文摘通过远缘杂交及回交将白菜的抗病基因导入到芥菜中,创制获得优质的抗根肿病长柄芥(Brassica juncea var.longepetiolata Yang et Chen)育种材料。采用人工接种鉴定和分子标记鉴定相结合法,对10份抗根肿病白菜材料进行抗性分析,筛选出1份可作为长柄芥抗根肿病育种材料创制的抗性资源CCR21002。利用CCR21002分别作为父本、母本提供抗根肿病基因,与长柄芥进行远缘杂交,结合胚挽救技术获得正反杂交种F1,选取植株形态偏向芥菜的F1与长柄芥回交获得BC_(1),对BC_(1)植株进行形态学、细胞学分析及抗病性鉴定。结果表明,回交一代比杂种一代更难获得,胚挽救成活的13株BC_(1)中,仅筛选出1株基因型为AABB且抗根肿病的植株,经分子标记鉴定所含抗病基因为CRb,在植株形态上保留有长柄芥的大部分性状。证明经过与抗根肿病白菜远缘杂交及回交,获得对4号生理小种表现抗性的长柄芥新种质1份,可用于继续回交转育。
