The somatic cell nuclear transfer （SCNT） technique has been applied successfully in a range of mammalian species giving rise to offspring, however, the efficiency of development to term has remained low. Oocyte meio...The somatic cell nuclear transfer （SCNT） technique has been applied successfully in a range of mammalian species giving rise to offspring, however, the efficiency of development to term has remained low. Oocyte meiotic arrest is an important option to make the SCNT more flexible and increase the number of cloned embryos produced. This paper showed that the use of butyrolactone I to arrest the meiotic division for 24 h prior to in vitro maturation （IVM） provided bovine oocytes capability of supporting development of blastocysts as efficiently as non arrested oocytes. In the SCNT group, cleavage rates were not affected by prematuration when adding butyrolactone I during IVM （57.9% vs. 62.4%; control vs. 10 μmol.L^-1 butyrolactone I）, developmental rates to biastocyst were unaffected （22.0% vs. 20.3%; control vs. 10 μmol.L^-1 butyrolactone I, p〉0.05） by the addition ofbutyrolactone I during IVM in cloned embryos. Moreover, the total cell numbers of blastocyst were not affected by butyrolactone I （total nucleus numbers, 132±16.5 vs. 128±19.4, p〉0.05）. In conclusion, the SCNT embryos from butyrolactone I- prematured bovine oocytes had the same developmental potential as the non treated one. The present study provided a method for laboratories of in vitro embryos production, mainly those working on the SCNT, where prematuration could be used to iiacrease the flexibility of the procedure.展开更多
为了建立水禽细小病毒(WPV)快速检测方法,根据序列比对结果在水禽细小病毒NS基因SF3保守区域内设计特异性引物,建立SYBR Green Ⅰ荧光定量PCR通用检测方法。该方法的扩增效率(E)为90.0%,相关系数(R~2)=0.99,标准曲线方程为y=-3.607x+38....为了建立水禽细小病毒(WPV)快速检测方法,根据序列比对结果在水禽细小病毒NS基因SF3保守区域内设计特异性引物,建立SYBR Green Ⅰ荧光定量PCR通用检测方法。该方法的扩增效率(E)为90.0%,相关系数(R~2)=0.99,标准曲线方程为y=-3.607x+38.77;除WPV出现S形扩增曲线外,新城疫病毒(NDV)、H9亚型禽流感病毒(H9 AIV)、鸭坦布苏病毒(DTMUV)、鸭肝炎病毒(DHAV)、鸭肠炎病毒(DEV)、鸭呼肠孤病毒(DRV)样品均未出现S形阳性扩增曲线;批内变异系数(CV)为0.15%~0.23%,批间变异系数为0.09%~0.28%。结果表明,SYBR Green Ⅰ荧光定量PCR检测方法重复性好、灵敏度高和特异性强。临床样品检测结果表明,SYBR Green Ⅰ荧光定量PCR与普通PCR的符合率达98.4%,灵敏度是普通PCR的1 000倍。SYBR Green Ⅰ荧光定量PCR检测方法不仅能定性检测WPV,还可以进行定量检测,可用于种鸭场、种鹅场的WPV净化检测,也可用于WPV临床大量样品的快速检测。展开更多
基金Supported by the Scientific Research Fund of Heilongjiang Provincial Education Department (11531028)Heilongjiang Postdoctoral Fund (LBH-Z11245)
文摘The somatic cell nuclear transfer （SCNT） technique has been applied successfully in a range of mammalian species giving rise to offspring, however, the efficiency of development to term has remained low. Oocyte meiotic arrest is an important option to make the SCNT more flexible and increase the number of cloned embryos produced. This paper showed that the use of butyrolactone I to arrest the meiotic division for 24 h prior to in vitro maturation （IVM） provided bovine oocytes capability of supporting development of blastocysts as efficiently as non arrested oocytes. In the SCNT group, cleavage rates were not affected by prematuration when adding butyrolactone I during IVM （57.9% vs. 62.4%; control vs. 10 μmol.L^-1 butyrolactone I）, developmental rates to biastocyst were unaffected （22.0% vs. 20.3%; control vs. 10 μmol.L^-1 butyrolactone I, p〉0.05） by the addition ofbutyrolactone I during IVM in cloned embryos. Moreover, the total cell numbers of blastocyst were not affected by butyrolactone I （total nucleus numbers, 132±16.5 vs. 128±19.4, p〉0.05）. In conclusion, the SCNT embryos from butyrolactone I- prematured bovine oocytes had the same developmental potential as the non treated one. The present study provided a method for laboratories of in vitro embryos production, mainly those working on the SCNT, where prematuration could be used to iiacrease the flexibility of the procedure.
文摘工程结构在制造工艺过程中或使用期间会产生裂纹,对结构断裂路径的预测和研究是防治工程安全问题发生的重要手段。在考虑裂纹尖端应力场常数项T应力的基础上对传统的最大周向应力准则(Maximum tangential stress criterion,MTS)和最小应变能密度因子准则(Minimum strain energy density criterion,SED)进行修正,采用Python语言对ABAQUS的前、后处理和有限元计算模块进行二次开发,通过计算最优解的粒子群算法(Particle swarm optimization,PSO)将修正后的准则编入裂纹自动扩展程序脚本中。利用上述二次开发程序对初始纯Ⅰ型裂纹的扩展路径进行模拟,结果表明:采用ABAQUS脚本程序模拟结果与相关文献实验结果吻合,表明了程序的有效性,进而实现考虑T应力的多种断裂准则对裂纹扩展路径的预测;当T应力值处于一定范围内时,修正的MTS准则无法预测裂纹发生的偏转现象,扩展路径呈直线,此时可采用修正的SED准则进行预测。
文摘为了建立水禽细小病毒(WPV)快速检测方法,根据序列比对结果在水禽细小病毒NS基因SF3保守区域内设计特异性引物,建立SYBR Green Ⅰ荧光定量PCR通用检测方法。该方法的扩增效率(E)为90.0%,相关系数(R~2)=0.99,标准曲线方程为y=-3.607x+38.77;除WPV出现S形扩增曲线外,新城疫病毒(NDV)、H9亚型禽流感病毒(H9 AIV)、鸭坦布苏病毒(DTMUV)、鸭肝炎病毒(DHAV)、鸭肠炎病毒(DEV)、鸭呼肠孤病毒(DRV)样品均未出现S形阳性扩增曲线;批内变异系数(CV)为0.15%~0.23%,批间变异系数为0.09%~0.28%。结果表明,SYBR Green Ⅰ荧光定量PCR检测方法重复性好、灵敏度高和特异性强。临床样品检测结果表明,SYBR Green Ⅰ荧光定量PCR与普通PCR的符合率达98.4%,灵敏度是普通PCR的1 000倍。SYBR Green Ⅰ荧光定量PCR检测方法不仅能定性检测WPV,还可以进行定量检测,可用于种鸭场、种鹅场的WPV净化检测,也可用于WPV临床大量样品的快速检测。
