目的:运用CRISPR/Cas9基因编辑技术构建前蛋白转化酶枯草溶菌素9(PCSK9)点突变家兔模型。方法:根据Pub Med基因蛋白数据对人和兔的PCSK9蛋白功能区进行Blast分析,发现人PCSK9基因的386S(丝氨酸)氨基酸功能区与兔PCSK9基因的485S同源。...目的:运用CRISPR/Cas9基因编辑技术构建前蛋白转化酶枯草溶菌素9(PCSK9)点突变家兔模型。方法:根据Pub Med基因蛋白数据对人和兔的PCSK9蛋白功能区进行Blast分析,发现人PCSK9基因的386S(丝氨酸)氨基酸功能区与兔PCSK9基因的485S同源。根据家兔PCSK9基因的485S对应的碱基替换位置及序列分析结果设计3条单链向导RNA和1条单链寡核苷酸供体模板。将合成的单链向导RNA、Cas9 m RNA和单链寡核苷酸供体共同注射入家兔受精卵细胞质内并将胚胎移植入待孕母兔体内。对获得的F0代兔进行PCR、TA克隆、脱靶检测以鉴定PCSK9S386A是否突变成功。利用获得的PCSK9S386A基因点突变家兔进行繁殖,扩大群体。结果:共获得15只F0代兔,其中1只为PCSK9S386A点突变纯合子,2只为PCSK9S386A点突变杂合子,且该突变可以稳定遗传。结论:利用CRISPR/Cas9技术成功构建了PCSK9S386A点突变家兔模型,为探究PCSK9功能减弱的分子机制,开发可靠、有效的诊断和治疗措施提供了良好的动物模型。展开更多
Mannitol is the first choice of clinical dehydrating agent, which can alleviate brain edema and reduce intracranial pressure. As a hypertonic solution, it has strong vascular irritation. Moreover, fast infusion speed ...Mannitol is the first choice of clinical dehydrating agent, which can alleviate brain edema and reduce intracranial pressure. As a hypertonic solution, it has strong vascular irritation. Moreover, fast infusion speed is required, which can be related to pain, physic liquor exomosis and some other side effects. Regarding of the physic liquor exosmosis in mannitol intravenous infusion, this paper reviewed the research progress of different external application treatment, and provided references for clinical treatment of physic liquor exosmosis in mannitol intravenous infusion.展开更多
Disruption of microtubule cytoskeleton plays an important role during the evolution of brain damage after transient cerebral ischemia.However,it is still unclear whether microtubule-stabilizing drugs such asepothilone...Disruption of microtubule cytoskeleton plays an important role during the evolution of brain damage after transient cerebral ischemia.However,it is still unclear whether microtubule-stabilizing drugs such asepothilone D(EpoD)have a neuroprotective action against the ischemia-induced brain injury.展开更多
Transient global ischemia usually results in delayed neuronal death in selective brain regions,prior to which a rapid loss of dendritic spines has been widely reported in these regions.Dendritic spines are characteriz...Transient global ischemia usually results in delayed neuronal death in selective brain regions,prior to which a rapid loss of dendritic spines has been widely reported in these regions.Dendritic spines are characterized by a highly branched meshwork of actin cytoskeleton(F-actin),which is extremely vulnerable to the ATP-depleted conditions such as hypoxia/ischemia.However,the ischemia-induced changes of F-actin are still not clarified in the vulnerable brain areas.This study was designed to examine the temporal and spatial alterations of F-actin in the CA1 subfield of rat hippocampus following reperfusion after global cerebral ischemia.展开更多
文摘目的:运用CRISPR/Cas9基因编辑技术构建前蛋白转化酶枯草溶菌素9(PCSK9)点突变家兔模型。方法:根据Pub Med基因蛋白数据对人和兔的PCSK9蛋白功能区进行Blast分析,发现人PCSK9基因的386S(丝氨酸)氨基酸功能区与兔PCSK9基因的485S同源。根据家兔PCSK9基因的485S对应的碱基替换位置及序列分析结果设计3条单链向导RNA和1条单链寡核苷酸供体模板。将合成的单链向导RNA、Cas9 m RNA和单链寡核苷酸供体共同注射入家兔受精卵细胞质内并将胚胎移植入待孕母兔体内。对获得的F0代兔进行PCR、TA克隆、脱靶检测以鉴定PCSK9S386A是否突变成功。利用获得的PCSK9S386A基因点突变家兔进行繁殖,扩大群体。结果:共获得15只F0代兔,其中1只为PCSK9S386A点突变纯合子,2只为PCSK9S386A点突变杂合子,且该突变可以稳定遗传。结论:利用CRISPR/Cas9技术成功构建了PCSK9S386A点突变家兔模型,为探究PCSK9功能减弱的分子机制,开发可靠、有效的诊断和治疗措施提供了良好的动物模型。
基金National Natural Science Foundation of China(81570392)National Key R&D Program of China(2016YFE0126000)
文摘Mannitol is the first choice of clinical dehydrating agent, which can alleviate brain edema and reduce intracranial pressure. As a hypertonic solution, it has strong vascular irritation. Moreover, fast infusion speed is required, which can be related to pain, physic liquor exomosis and some other side effects. Regarding of the physic liquor exosmosis in mannitol intravenous infusion, this paper reviewed the research progress of different external application treatment, and provided references for clinical treatment of physic liquor exosmosis in mannitol intravenous infusion.
文摘Disruption of microtubule cytoskeleton plays an important role during the evolution of brain damage after transient cerebral ischemia.However,it is still unclear whether microtubule-stabilizing drugs such asepothilone D(EpoD)have a neuroprotective action against the ischemia-induced brain injury.
文摘Transient global ischemia usually results in delayed neuronal death in selective brain regions,prior to which a rapid loss of dendritic spines has been widely reported in these regions.Dendritic spines are characterized by a highly branched meshwork of actin cytoskeleton(F-actin),which is extremely vulnerable to the ATP-depleted conditions such as hypoxia/ischemia.However,the ischemia-induced changes of F-actin are still not clarified in the vulnerable brain areas.This study was designed to examine the temporal and spatial alterations of F-actin in the CA1 subfield of rat hippocampus following reperfusion after global cerebral ischemia.
