湿热证大鼠肾内髓及尿液中水通道蛋白AQP2含量的变化

目的 :探讨湿热证大鼠肾内髓AQP2和尿液AQP2与湿热证中“湿”的形成的关系。方法 :复制湿热证大鼠模型 ,检测各组肾内髓及尿液AQP2的含量。结果 :湿热证模型组AQP2水平较正常组明显降低 ,尿液AQP2蛋白的排出量与肾脏AQP2蛋白的表达量有着良好的相关性。结论 :模型组大鼠肾脏AQP2的减少与湿热因素造模有着密切联系 ,尿液AQP2含量可以反映肾内髓AQP2蛋白的水平。

Management of surgical splenorenal shunt-related hepatic myelopathy with endovascular interventional techniques

We present a case with hepatic myelopathy(HM) due to a surgical splenorenal shunt that was successfully treated by endovascular interventional techniques.A 39-year-old man presented with progressive spastic paraparesis of his lower limbs 14 mo after a splenorenal shunt.A portal venogram identified a widened patent splenorenal shunt.We used an occlusion balloon catheter initially to occlude the shunt.Further monitoring of the patient revealed a decrease in his serum ammonia level and an improvement in leg strength.We then used an Amplatzer vascular plug(AVP) to enable closure of the shunt.During the follow up period of 7 mo,the patient experienced significant clinical improvement and normalization of blood ammonia,without any complications.Occlusion of a surgically created splenorenal shunt with AVP represents an alternative therapy to surgery or coil embolization that can help to relieve shunt-induced HM symptoms.

利用常间回文重复序列丛集及相关蛋白9系统敲除小鼠肾集合管上皮细胞系的水通道蛋白2基因

目的利用常间回文重复序列丛集(CRISPR)/CRISPR相关蛋白9(Cas9)系统敲除小鼠肾内髓集合管3(m-IMCD3)上皮细胞的水通道蛋白2(Aqp2)基因。方法在小鼠Aqp2基因的第1、4个外显子上各设计两个小向导RNA(sgRNA),分别为sgRNA1-1、sgRNA1-2、sgRNA4-1、sgRNA4-2,并将其成功克隆进常间回文重复序列丛集慢病毒载体2上。将测序正确的质粒转染到m-IMCD3细胞中,提取各单克隆细胞系的基因组DNA,通过聚合酶链反应扩增出sgRNA作用靶点的DNA片段并测序。利用逆转录聚合酶链反应(RT-PCR)及免疫荧光检测Aqp2的表达。结果成功构建出含有相应sgRNA的载体;4个sgRNA对Aqp2基因的靶点都有切割作用;sgRNA1-1与sgRNA4-2组合能成功把两者间5500bp的DNA片段敲除;应用RT-PCR及免疫荧光证实应用CRISPR/Cas9系统敲除Aqp2后,该m-IMCD3细胞系中的Aqp2mRNA及蛋白几乎未见表达。结论通过CRISPR/Cas9系统可获得Aqp2基因敲除的肾集合管上皮细胞系。