目的建立稳定表达CRFR1的HEK293细胞株,并鉴定cAMP评价体系构建是否成功。方法采用Lipofectamine2000将CRFR1质粒转染至HEK293细胞中,经G418筛选单克隆阳性细胞,采用Western blot技术、RT-PCR法、免疫荧光法证实稳定表达CRFR1细胞株构...目的建立稳定表达CRFR1的HEK293细胞株,并鉴定cAMP评价体系构建是否成功。方法采用Lipofectamine2000将CRFR1质粒转染至HEK293细胞中,经G418筛选单克隆阳性细胞,采用Western blot技术、RT-PCR法、免疫荧光法证实稳定表达CRFR1细胞株构建成功。并用CRF刺激稳定表达CRFR1的HEK293细胞株,绘制CRF刺激HEK293-CRFR1细胞释放cAMP的量效曲线。结果 Western blot、RT-PCR、免疫荧光结果表明,CRFR1受体在HEK293细胞系中成功表达。cAMP释放量效实验显示,CRF刺激HEK293-CRFR1细胞释放cAMP的EC50为(5.64±0.05)×10-10mol·L-1。结论成功建立了稳定表达CRFR1的HEK293细胞株,并且cAMP含量评价体系构建成功,为研究CRFR1的生物学功能及筛选CRFR1受体靶向药物奠定了基础。展开更多
Modelling and biomedical applications of ultrasound contrast agent （UCA） microbubbles have attracted a great deal of attention. In this review, we summarize a series of researches done in our group, including （i） ...Modelling and biomedical applications of ultrasound contrast agent （UCA） microbubbles have attracted a great deal of attention. In this review, we summarize a series of researches done in our group, including （i） the development of an all-in-one solution of characterizing coated bubble parameters based on the light scattering technique and flow cytometry; （ii） a novel bubble dynamic model that takes into consideration both nonlinear shell elasticity and viscosity to eliminate the dependences of bubble shell parameters on bubble size; （iii） the evaluation of UCA inertial cavitation threshold and its relationship with shell parameters; and （iv） the investigations of transfection efficiency and the reduction of cytotoxicity in gene delivery facilitated by UCAs excited by ultrasound exposures.展开更多
文摘目的建立稳定表达CRFR1的HEK293细胞株,并鉴定cAMP评价体系构建是否成功。方法采用Lipofectamine2000将CRFR1质粒转染至HEK293细胞中,经G418筛选单克隆阳性细胞,采用Western blot技术、RT-PCR法、免疫荧光法证实稳定表达CRFR1细胞株构建成功。并用CRF刺激稳定表达CRFR1的HEK293细胞株,绘制CRF刺激HEK293-CRFR1细胞释放cAMP的量效曲线。结果 Western blot、RT-PCR、免疫荧光结果表明,CRFR1受体在HEK293细胞系中成功表达。cAMP释放量效实验显示,CRF刺激HEK293-CRFR1细胞释放cAMP的EC50为(5.64±0.05)×10-10mol·L-1。结论成功建立了稳定表达CRFR1的HEK293细胞株,并且cAMP含量评价体系构建成功,为研究CRFR1的生物学功能及筛选CRFR1受体靶向药物奠定了基础。
基金Projects supported by the National Natural Science Foundation of China(Grant Nos.81127901,81227004,11374155,11274170,11274176,11474001,11474161,11474166,and 11674173)the National High-Technology Research and Development Program,China(Grant No.2012AA022702)Qing Lan Project of Jiangsu Province,China
文摘Modelling and biomedical applications of ultrasound contrast agent （UCA） microbubbles have attracted a great deal of attention. In this review, we summarize a series of researches done in our group, including （i） the development of an all-in-one solution of characterizing coated bubble parameters based on the light scattering technique and flow cytometry; （ii） a novel bubble dynamic model that takes into consideration both nonlinear shell elasticity and viscosity to eliminate the dependences of bubble shell parameters on bubble size; （iii） the evaluation of UCA inertial cavitation threshold and its relationship with shell parameters; and （iv） the investigations of transfection efficiency and the reduction of cytotoxicity in gene delivery facilitated by UCAs excited by ultrasound exposures.
