1994年MacManus等首先报告大鼠全脑缺血后存在神经元凋亡。近10年来，关于脑缺血后细胞凋亡机制的研究取得了巨大进展。已知脑缺血后可有多种基因被诱导表达，这些基因编码的蛋白质产物直接或间接参与了凋亡的调控。天冬氨酸特异性半胱...1994年MacManus等首先报告大鼠全脑缺血后存在神经元凋亡。近10年来，关于脑缺血后细胞凋亡机制的研究取得了巨大进展。已知脑缺血后可有多种基因被诱导表达，这些基因编码的蛋白质产物直接或间接参与了凋亡的调控。天冬氨酸特异性半胱氨酸蛋白酶家族（cysteinyl aspartate specific protease，caspase），是细胞凋亡过程中最重要的蛋白酶，细胞凋亡的最后实施是通过caspase的激活而实现的。迄今为止，已知的家族成员共14个，其中caspase-8和caspase-9在细胞凋亡启动过程中具有重要作用。在本实验中，我们观察了大鼠局灶性脑缺血-再灌注后神经元凋亡及caspase-8、caspase-9 mRNA和蛋白质表达的变化，以探讨脑缺血-再灌注后神经元凋亡的信号转导机制。展开更多
目的观察温针灸内、外膝眼穴对膝骨性关节炎兔软骨组织细胞调亡调控蛋白Caspase-3、Caspase-9表达的调节作用。方法将18只新西兰兔随机分为正常对照组(正常组)、膝骨性关节炎组(模型组)和温针内、外膝眼穴组(温针组)。采用关节腔注射木...目的观察温针灸内、外膝眼穴对膝骨性关节炎兔软骨组织细胞调亡调控蛋白Caspase-3、Caspase-9表达的调节作用。方法将18只新西兰兔随机分为正常对照组(正常组)、膝骨性关节炎组(模型组)和温针内、外膝眼穴组(温针组)。采用关节腔注射木瓜蛋白酶法制作膝骨性关节炎模型。造模后温针组于内外膝眼穴处行温针灸20min,隔天1次,每2周为1个疗程,中间休息2d,共治疗4个疗程。模型组和正常组不给予任何治疗。RT-q PCR法检测软骨组织Caspase-3、Caspase-9 m RNA表达。结果 (1)温针灸干预后,新西兰兔的精神、食欲、膝关节活动能力等均明显改善。(2)模型组与正常组比较,膝关节软骨组织Caspase-3、Caspase-9 m RNA表达显著升高(P<0.001),温针组与模型组比较,膝关节软骨组织Caspase-3、Caspase-9 m RNA表达显著下调(P<0.05)。结论温针灸内、外膝眼穴能下调膝关节软骨组织中细胞调亡调控蛋白Caspase-3、Caspase-9 m RNA的表达,抑制软骨细胞凋亡通路的信号转导。展开更多
CRISPR/Cas9 technology is a powerful genome manipulation tool in insects.However,little is known about whether mRNA and protein of a target gene are completely cleared in homozygous mutants.This study generated homozy...CRISPR/Cas9 technology is a powerful genome manipulation tool in insects.However,little is known about whether mRNA and protein of a target gene are completely cleared in homozygous mutants.This study generated homozygous mutants of the insulin receptor gene 2(NlInR2)in the brown planthopper(Nilaparvata lugens)using CRISPR/Cas9 genome editing.Both frameshift mutants,E5_D17 and E6_I7,differentiated towards long wings,but there were differences in wing morphology,with E5_D17 showing wing deformities.Subsequent investigations revealed the presence of residual expression of NlInR2 mRNA in both mutants,as well as the occurrence of spliceosomes featuring exon skipping splicing in E5_D17.Additionally,the E5_D17 exhibited the detection of N-terminally truncated NlInR2 protein.RNA interference experiments indicated that the knockdown of NlInR2 expression in the E5_D17 mutant line increased the proportion of wing deformities from 11.1 to 65.6%,suggesting that the residual NlInR2 mRNA of the E5_D17 mutant might have retained some genetic functions.Our results imply that systematic characterization of residual protein expression or function in CRISPR/Cas9-generated mutant lines is necessary for phenotypic interpretation.展开更多
文摘1994年MacManus等首先报告大鼠全脑缺血后存在神经元凋亡。近10年来，关于脑缺血后细胞凋亡机制的研究取得了巨大进展。已知脑缺血后可有多种基因被诱导表达，这些基因编码的蛋白质产物直接或间接参与了凋亡的调控。天冬氨酸特异性半胱氨酸蛋白酶家族（cysteinyl aspartate specific protease，caspase），是细胞凋亡过程中最重要的蛋白酶，细胞凋亡的最后实施是通过caspase的激活而实现的。迄今为止，已知的家族成员共14个，其中caspase-8和caspase-9在细胞凋亡启动过程中具有重要作用。在本实验中，我们观察了大鼠局灶性脑缺血-再灌注后神经元凋亡及caspase-8、caspase-9 mRNA和蛋白质表达的变化，以探讨脑缺血-再灌注后神经元凋亡的信号转导机制。
文摘目的观察温针灸内、外膝眼穴对膝骨性关节炎兔软骨组织细胞调亡调控蛋白Caspase-3、Caspase-9表达的调节作用。方法将18只新西兰兔随机分为正常对照组(正常组)、膝骨性关节炎组(模型组)和温针内、外膝眼穴组(温针组)。采用关节腔注射木瓜蛋白酶法制作膝骨性关节炎模型。造模后温针组于内外膝眼穴处行温针灸20min,隔天1次,每2周为1个疗程,中间休息2d,共治疗4个疗程。模型组和正常组不给予任何治疗。RT-q PCR法检测软骨组织Caspase-3、Caspase-9 m RNA表达。结果 (1)温针灸干预后,新西兰兔的精神、食欲、膝关节活动能力等均明显改善。(2)模型组与正常组比较,膝关节软骨组织Caspase-3、Caspase-9 m RNA表达显著升高(P<0.001),温针组与模型组比较,膝关节软骨组织Caspase-3、Caspase-9 m RNA表达显著下调(P<0.05)。结论温针灸内、外膝眼穴能下调膝关节软骨组织中细胞调亡调控蛋白Caspase-3、Caspase-9 m RNA的表达,抑制软骨细胞凋亡通路的信号转导。
基金the National Natural Science Foundation of China(31730073).
文摘CRISPR/Cas9 technology is a powerful genome manipulation tool in insects.However,little is known about whether mRNA and protein of a target gene are completely cleared in homozygous mutants.This study generated homozygous mutants of the insulin receptor gene 2(NlInR2)in the brown planthopper(Nilaparvata lugens)using CRISPR/Cas9 genome editing.Both frameshift mutants,E5_D17 and E6_I7,differentiated towards long wings,but there were differences in wing morphology,with E5_D17 showing wing deformities.Subsequent investigations revealed the presence of residual expression of NlInR2 mRNA in both mutants,as well as the occurrence of spliceosomes featuring exon skipping splicing in E5_D17.Additionally,the E5_D17 exhibited the detection of N-terminally truncated NlInR2 protein.RNA interference experiments indicated that the knockdown of NlInR2 expression in the E5_D17 mutant line increased the proportion of wing deformities from 11.1 to 65.6%,suggesting that the residual NlInR2 mRNA of the E5_D17 mutant might have retained some genetic functions.Our results imply that systematic characterization of residual protein expression or function in CRISPR/Cas9-generated mutant lines is necessary for phenotypic interpretation.
