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Construction and characterization of a synthesized herpes simplex virus H129-Syn-G2 被引量:1
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作者 Han Xiao Hengrui Hu +6 位作者 Yijia Guo Jiang Li Le Wen Wen-Bo Zeng Manli Wang Min-Hua Luo Zhihong Hu 《Virologica Sinica》 SCIE CAS CSCD 2023年第3期373-379,共7页
Herpes simplex virus type 1(HSV-1)causes lifelong infections worldwide,and currently there is no efficient cure or vaccine.HSV-1-derived tools,such as neuronal circuit tracers and oncolytic viruses,have been used exte... Herpes simplex virus type 1(HSV-1)causes lifelong infections worldwide,and currently there is no efficient cure or vaccine.HSV-1-derived tools,such as neuronal circuit tracers and oncolytic viruses,have been used exten-sively;however,further genetic engineering of HSV-1 is hindered by its complex genome structure.In the present study,we designed and constructed a synthetic platform for HSV-1 based on H129-G4.The complete genome was constructed from 10 fragments through 3 rounds of synthesis using transformation-associated recombination(TAR)in yeast,and was named H129-Syn-G2.The H129-Syn-G2 genome contained two copies of the gfp gene and was transfected into cells to rescue the virus.According to growth curve assay and electron microscopy results,the synthetic viruses exhibited more optimized growth properties and similar morphogenesis compared to the parental virus.This synthetic platform will facilitate further manipulation of the HSV-1 genome for the devel-opment of neuronal circuit tracers,oncolytic viruses,and vaccines. 展开更多
关键词 herpes simplex virus type 1(hSV-1) Neuronal circuit tracers h129-syn-g2 h129-G4 Synthetic biology
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MicroRNA-129-5p在糖尿病性心肌病中的作用 被引量:1
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作者 孔德忆 孔霄 《中国老年学杂志》 CAS 北大核心 2021年第23期5356-5359,共4页
目的探究微小RNA(miRNA)miR-129-5p在糖尿病心肌病中的表达及生物学功能。方法采用浓度为5.5 mmol/L葡萄糖(对照组)和50.0 mmol/L葡萄糖(高糖组)干预H9C2心肌细胞0、24、48、72、96 h后,采用实时荧光聚合酶链式反应(qRT-PCR)检测H9C2心... 目的探究微小RNA(miRNA)miR-129-5p在糖尿病心肌病中的表达及生物学功能。方法采用浓度为5.5 mmol/L葡萄糖(对照组)和50.0 mmol/L葡萄糖(高糖组)干预H9C2心肌细胞0、24、48、72、96 h后,采用实时荧光聚合酶链式反应(qRT-PCR)检测H9C2心肌细胞中miR-129-5p的表达;分别将miR-129-5p模拟物(HG+miR-129-5p模拟物组)及其模拟物阴性对照物(HG+mimics-NC组)转染至高糖诱导下的H9C2细胞中,并设置对照组和高糖组,采用酶联免疫吸附试验(ELISA)检测各组细胞上清液肿瘤坏死因子(TNF)-α、白细胞介素(IL)-1β、超氧化物歧化酶(SOD)和丙二醛(MDA)的相对水平,采用CCK-8检测各组细胞存活率,采用流式细胞仪检测H9C2心肌细胞的凋亡,采用免疫印迹检测各组细胞Ki67、Bax、酶切-caspase-3蛋白表达。结果与对照组相比,miR-129-5p在高糖组的H9C2心肌细胞中表达显著下调(P<0.05),且呈时间依赖关系。与对照组相比,高糖组H9C2心肌细胞中IL-1β、TNF-α和MDA的相对含量显著升高(P<0.01),而SOD相对含量显著降低(P<0.01);与高糖+mimics-NC组相比,高糖+miR-129-5p模拟物组H9C2心肌细胞中IL-1β、TNF-α和MDA相对含量显著降低(P<0.05),而SOD相对含量显著升高(P<0.01)。与对照组相比,高糖组H9C2心肌细胞凋亡率、Bax和酶切-caspase-3蛋白水平显著升高(P<0.001),而细胞存活率和Ki67蛋白显著降低(P<0.01);与高糖+mimics-NC组相比,高糖+miR-129-5p模拟物组中H9C2心肌细胞凋亡率、Bax和酶切-caspase-3蛋白水平显著降低(P<0.001),而细胞存活率和Ki67蛋白显著增加(P<0.01)。结论 miR-129-5p抑制高糖诱导的H9C2心肌细胞的炎症反应、氧化应激及凋亡,并促进增殖。 展开更多
关键词 微小RNA miR-129-5p 糖尿病心肌病 h9C2 凋亡
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