作者机构：
[周平坤; 岳小乔] The School of Public Health, University of South China, Hengyang, 421001, China;[谢达菲; 白琛俊; 刘晓丹] The Institute of Radiation Medicine, The Academy of Military Medical Science, Beijing, 100850, China
通讯机构：
[Zhou, P.] T;The School of Public Health, University of South China, Hengyang, China
作者机构：
[Xiuqin Y.] School of Public Health, University of South Ghina, Hengyang, 421001, China;[Zhiguang L.; Kanglin W.; Lili Z.; MacHao L.; Xiuqin Z.; Haican L.] State Key Laboratory for Infrctious Diseases Prevention and Control, Collaborative Innovation Center for Diagnosis and Treatment of Infrctious Diseases, National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing, 102206, China;[Yao L.] School of Laboratory Medicine and LJ Science, Iedical University, Ifenzhou, 325035, China;[Yunli D.; Rong C.; Zixin C.] School of Public Health, University of South Ghina, Hengyang, 421001, China, State Key Laboratory for Infrctious Diseases Prevention and Control, Collaborative Innovation Center for Diagnosis and Treatment of Infrctious Diseases, National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing, 102206, China
通讯机构：
[Xiuqin, Y.] S;School of Public Health, University of South Ghina, Hengyang, China
作者机构：
[Zeng, H C; 王利; 徐新云; 秦逍云; 王冰玉] Institue of Toxicology, Shenzhen Center for Disease Control and Prevention, Shenzhen 518055, China;[Zeng, H C; 龙鼎新; 王利; 王冰玉; 郑凯] The Public Sanitation College of Nanhua University, Hengyang 421001, China
作者：
Li Fei-fei;Lu Yi-song;Yang Sheng-yuan*;Lin Xi;Chen Wei;...
期刊：
光谱学与光谱分析,2019年39(9):2769-2773 ISSN：1000-0593
通讯作者：
Yang Sheng-yuan
作者机构：
[Li Fei-fei; Chen Wei; Yang Sheng-yuan; Lin Xi; Xiao Fu-bing; Liang Hao; Liu Can] Univ South China, Coll Publ Hlth, Hengyang 421001, Peoples R China.;[Lu Yi-song] Univ South China, Affiliated Nanhua Hosp, Hengyang 421001, Peoples R China.;[Li Fei-fei; Yang Sheng-yuan; Lin Xi; Xiao Fu-bing; Liang Hao; Liu Can] Key Lab Hengyang Hlth Hazard Factors Inspect & Qu, Hengyang 421001, Peoples R China.
通讯机构：
[Yang Sheng-yuan] U;[Yang Sheng-yuan] K;Univ South China, Coll Publ Hlth, Hengyang 421001, Peoples R China.;Key Lab Hengyang Hlth Hazard Factors Inspect & Qu, Hengyang 421001, Peoples R China.
关键词：
裂开型核酸适体;单壁碳纳米管;荧光
摘要：
基于裂开型核酸适体序列短、能有效降低因探针形成二级结构产生假阳性信号等优点,选择裂开型核酸适体作为特异性识别探针,核酸染料噻唑橙(TO)为信号探针,用单壁碳纳米管(SWCNTs)降低背景信号,利用“适配体-目标分子-适配体”的“三明治”夹心方式,建立了一种检测ATP的新方法。在pH 8.0的Tris-HCl缓冲溶液中,裂开成两段的ATP适体特异性识别ATP分子,生成稳定的“适配体-ATP-适配体”复合结构。单壁碳纳米管对该复合结构的吸附力较弱,因此该复合物游离在溶液中,TO与其结合而产生强荧光。当不存在ATP时,核酸适体探针以单链状态存在,可通过π—π共轭作用结合到SWCNTs表面,进而不能与TO结合,TO游离在溶液中荧光非常微弱。反应体系中ATP浓度越高,形成的“适配体-ATP-适配体”夹心识别结构复合物越多,检测到的荧光强度越大,据此实现对ATP的检测。在优化实验条件下,在最大荧光发射波长550nm处,ATP的浓度在9.0×10~(-9)~1.0×10~(-7) mol·L~(-1)范围内与ΔF/F0值成线性关系,r=0.996 4。该方法加标回收率为95.2%~104%,相对标准偏差(RSD)为1.02%~4.54%,检出限达到2.67×10~(-9) mol·L~(-1)。该方法基于功能核酸对目标物亲合力强、选择识别性高的特点,对ATP的检测表现出很好的选择性,实验结果表明,当相对误差控制在±5%以内时,200倍的UTP,GTP和CTP均不干扰 ATP的测定。另外,该方法操作简单、快速、无需标记、灵敏准确,可用于血清样品中ATP的测定,在快速检测小分子物质领域中有较好的应用前景。 <&wdkj&>A novel Label-Free Fluorometric Assay based on the recombination of split aptamer chip was developed for thedetection of adenosine triphosphate(ATP).In this strategy,the split aptamer was selected as a specific capture probe for the split two fragments aptamers could specifically form a ternary assembly in the presence of ligand and the two separate oligonucleotides lack secondary structures,thus not yielding false-positive or nonspecific signals,while the Thiazole orange(TO),an almost nonfluorescence dye in buffer solution,was used as signal probe,and the single-walled carbon nanotubes(SWCNTs)was applied to reduce the background signals.In the pH 8.0Tris-HCl buffer solution,those two split aptamer fragments will be combined with each other to form a stable“aptamer-ATP-aptamer”composite structure upon interacting with its target ATP.The“sandwich” structure can't wrap the sidewalls of the SWCNTs and is freed in solution,and TO shows agreat fluorescence enhancement when binding to the“aptamer-ATP-aptamer”composite structure.In the absence of ATP,the split aptamers,existing in a singlestranded state,bind to the surface of the SWCNTs via aπ—π-conjugate interaction,and TO shows weak fluorescence because “sandwich”structure is not formed.In the system,the higher the ATP concentration is,the more the“aptamer-ATP-aptamer” sandwich recognition structure complex obtained,sois the fluorescence.Under the optimized experimental conditions,the ATP concentration in the range from 9.0×10~(-9) mol·L~(-1) to 1.0×10~(-7) mol·L~(-1) was linear with theΔF/F0value at the maximum fluorescence emission wavelength of 550nm,r=0.996 4,with a low detection limit of 2.67×10~(-9) mol·L~(-1).The recoveries of the method were 95.2%~104%,and the relative standard deviation(RSD)was 1.02%~4.54%,respectively.Based on the specific molecular recognition and high affinity of twosplit aptamers,the reaction product was shown that a“turn-on”fluorescence response to ATP with good selectivity,only a slight fluorescence change could be observed by GTP,CTP,and UTP(at a 200-fold higher concentration than that of ATP),indicating that UTP,CTP,and GTP could not interact with P1and P1to initiate the reaction.The method is simple,rapid,free-label,sensitive and accurate,and can be used for the determination of ATP in serum samples.Therefore,the present strategy has a great potential application prospect in the field of rapid detection of small molecular substances.
作者机构：
[龙鼎新; 王冰玉; 郑凯] School of Public Health, South China University, Hengyang 421001, China;[徐新云; 王冰玉; 郑凯] Institue of Environment and Health, Shenzhen Center for Disease Control and Prevention, Shenzhen 518055, China;[耿红] Institute of Environmental Science, Shanxi University, Taiyuan 030006, China
作者机构：
[牛磊; 刘政海; 万炜; 曹文宇; 罗诗诗; 王贞] Clinical Anatomy & Reproductive Medicine Application Institute, Medical College, University of South China, Hengyang, 421001, China;[乃爱桃] Department of Radiation Oncology, The First Affiliated Hospital of University of South China, Hengyang, 421001, China;[徐杨] Department of Physiology, Medical College, University of South China, Hengyang, 421001, China;[何洁] Department of Pathology, Medical College, University of South China, Hengyang, 421001, China;[何淑雅] Department of Radiation Medicine, School of Public Health, University of South China, Hengyang, 421001, China
通讯机构：
[Wan, W.] C;Clinical Anatomy & Reproductive Medicine Application Institute, China
关键词：
电离辐射;海马;星形胶质细胞;认知功能障碍
摘要：
探讨电离辐射引起小鼠认知功能障碍的可能机制。24只雌性昆明小鼠随机分为对照组和辐照组,辐照组小鼠接受137Csγ射线单次全身辐照至吸收剂量4 Gy。35 d后采用Y迷宫实验检测小鼠认知功能;免疫组织化学法检测小鼠海马区星形胶质细胞(Astrocytes,Ast)标记物胶质纤维酸性蛋白(Glial fibrillary acidic protein,GFAP)阳性细胞数的表达变化;Western blot方法检测海马GFAP蛋白的表达变化;荧光定量PCR检测炎性因子白细胞介素IL-1α、IL-1β、IL-6 mRNA和海马GFAP的表达情况。结果表明:与对照组相比,辐照组小鼠在Y迷宫实验中自发交替率明显降低(78.40±4.16 vs. 67.53±3.08,p<0.05);海马区GFAP阳性细胞数明显增加(133.4±9.6 vs. 280.3±32.7,p<0.01);GFAP蛋白及mRNA表达均显著上调(1.000±0.036vs. 1.300±0.102,p<0.05;1.000±0.031 vs. 1.203±0.078,p<0.05);IL-1α、IL-1β和IL-6 mRNA的表达均明显上调(1.000±0.133 vs. 1.677±0.164,p<0.01;1.000±0.132 vs. 1.488±0.105,p<0.05;1.000±0.218 vs.2.181±0.188,p<0.01)。研究提示,电离辐射可能是通过激活海马Ast诱导炎症因子释放,从而引起认知功能障碍。
