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Temperature-sensitive goldnanotube array membranes modified with poly(N-isopropylacrylamide)

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Chinese Science Bulletin 2008年第5期53卷 727-732页

作者： yang xiaohai WU YingBen WANG Qing WANG KeMin WANG ShengFeng Biomedical Engineering Center State Key Laboratory of Chemo/Biosensing and Chemometrics College of Chemistry and ChemicalEngineering Key Laboratory for Bio-Nanotechnology and Molecular Engineering of Hunan Province Hunan University Changsha 410082 China

A kind of temperature-sensitive nanotube array membrane was developed by modifying gold-nanotube array membranes with poly(N-isopropylacrylamide) (PNIPAm). The permeation ability of the mem-branes at different temperatures was investigated using sodium fluorescein and quantum dots as probes. The results showed that the pore diameter of nanotube was changed due to the reversible re-sponse of PNIPAm-modified membranes to temperature, and then the permeation ability of the mem-branes was changed. The permeation of fluorescence probes was slow and even almost blocked at 25℃ (below the lower critical solution temperature, LCST), since PNIPAm formed expanded structures and decreased the pore size. While at 40℃ (above the LCST), the permeation was increased, since PNIPAm became compact structures and the pore diameter was increased. Furthermore, the permea-tion ability of the temperature-sensitive nanotube array membranes could be adjusted reversibly and it is possible to use the membranes in nanofluidic devices, nanogates, etc.

关键词：金纳米管阵列纳米薄膜温度敏感性纳米技术

mRNA detection in living cell using phosphorothioate-modified molecular beacon

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Chinese Science Bulletin 2009年第9期54卷 1507-1514页

作者： TANG HongXing yang xiaohai WANG KeMin TAN WeiHong LI Wei State Key Laboratory of Chemo/Biosensing and Chemometrics College of Chemistry and Chemical Engineering Biomedical Engineering Center and Key Laboratory for Bio-Nanotechnology and Molecular Engineering of Hunan Province Hunan University Changsha 410082 China

In this study, GFP mRNA in COS-7 cell and GFP-transfected COS-7 cell was detected in real time using phosphorothioate-modified molecular beacon based on living cell imaging method. Results showed that phosphorothioate-modified molecular beacon still kept the advantages of molecular beacon, such as, excellent selectivity, high sensitivity, and no separation detection. In addition, this modification could significantly increase the nuclease resistance of molecular beacon. Phosphorothioate-modified molecular beacon can efficiently reduce the false positive signal and improve the accuracy of living cell mRNA detection.

关键词：分子信标分离检测 mRNA 活细胞灯塔绿色荧光蛋白基因 ph 基因转染

A novel fluorescent label based on biological fluores-cent nanoparticles and its application in cell recognition

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Chinese Science Bulletin 2001年第23期46卷 1962-1965,2025页

作者： Xiaoxiao He Kemin Wang Weihong Tan Dan Xiao Jun Li xiaohai yang Hunan Univ Coll Chem & Chem Engn Changsha 410082 Peoples R China

Uniform-sized fluorescent nanoparticles have been prepared by employing silica as the shell and a highly luminescent dye complex of ruthenium ion and bipyridyl, tris(2,2 ’-bipyridyl) dichlororuthenium( II) hexahydrate as the core of the nanoparticles. A novel fluorescent label method is proposed, which is based on the biological fluorescent nanoparticles on the foundation of nanotechnology, biotechnology and fluorescent label technology. In comparison with the conventional fluorophores as fluorescent labels such as fluorescein isothiocyanate (FITC) label, this new label shows more superiority in photochemical stability, detection sensitivity and application scope for the biomedicine research. SmIgG+ B lymphocytes isolated from the circulating blood of human beings can be easily recognized by using this new fluorescent label.

关键词： biological fluorescent nanoparticles fluorescent label cell recognition.

Biomimetic nanochannel membrane for cascade response of borate and cis-hydroxyl compounds:An IMP logic gate device

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Chinese Chemical Letters 2019年第7期30卷 1397-1400页

作者： Meitao Deng Meng yang Yaqing Xu Yuqiong Sun Qing Wang Jianbo Liu Jin Huang xiaohai yang Kemin Wang State Key Laboratory of Chemo/Biosensing and Chemometrics College of Chemistry and Chemical Engineering Key Laboratory for Bio-Nanotechnology and Molecular Engineering of Hunan Province Hunan University Changsha 410082 China College of Chemistry Biology and Environment Yuxi Normal University Yuxi 653100 China

Biomimetic nanodevice is an important branch to expand the potential applications of artificial nanochannels. Here, we constructed nanochannels for cascade response of borate and cis-hydroxyl compounds by modifying the nanochannels of track-etched polycarbonate (TEPC) membranes with polyvinyl alcohol (PVA). Firstly, borate bound to PVA and increased the negative charge density on the surface of the nanochannels, which obstructed the transport of 1,5-naphthalene disulfonate (NDS2-). Subsequently, the addition of cis-hydroxyl compounds induced leaving of borate due to the stronger binding affinities between borate and cis-hydroxyl compounds, which reduced the negative charge density on nanochannels and then enhanced the transport of NDS2-. The cascade response of the nanochannels also accord with the properties of IMP (implication) logic gate. In addition, such nanochannels showed good reproducibility and reversibility. Therefore, this cascade response model based on nanochannels has the potential to be used as switches in area of actuators and biosensors, and is also expected to be used to understand the interaction of substances in nanoscale and simulate the physiological functions of boron.

关键词： Biomimetic nanochannel Cascade response Logic gate Borate cis-Hydroxyl compound

Engineering self-catabolic DNAzyme nanospheres for synergistic anticancer therapy

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Science China Chemistry 2024年第7期67卷 2412-2422页

作者： Yu Chen Yu Guo Jiaoli Wang Ruiting Liu xiaohai yang Kemin Wang Ying Pu Hui Shi Jin Huang Key Laboratory of Chemo/Biosensing and Chemometrics College of Chemistry and Chemical EngineeringKey Laboratory for Bio-Nanotechnology and Molecular Engineering of Hunan ProvinceHunan UniversityChangsha 410082China National Clinical Research Center for Geriatric Disorders Department of GeriatricsXiangya HospitalCentral South UniversityChangsha 410008China

DNAzyme-based gene therapy faces some challenges including cell penetration,activity limitation,and co-delivery ***-assembled DNA nanomedicine has attracted widespread attention due to its many *** is urgent to develop a universal DNA degradation strategy for precise programmable drug ***,we reported a self-catabolic DNAzyme nanospheres(SCNS),which could simultaneously achieve cell penetration,activity enhancement,and co-delivery *** SCNS were assembled through Y-DNA stepwise hybridization with each other,which were then loaded with aptamer(Apt),doxorubicin(Dox),and zinc oxide nanoparticles(ZnO NPs).The acid-triggered dissociation of ZnO NPs leads to the generation of Zn^(2+)ions cofactors for immediately self-catabolic DNAzyme *** the disassembly of the SCNS,three types of anticancer treatments would be activated,which include Zn^(2+)involved reactive oxygen species(ROS),Dox-induced chemotherapy,and DNAzyme-based gene *** experimental results show that the nanoplatform(Apt-SCNS-Dox-ZnO)has a good tumor-killing effect and minimal side *** a smart self-driven drug delivery nanoplatform,it is anticipated to displace extraordinary potential in biomedicine and bioengineering.

关键词： DNAzyme nanosphere ZnO NPs ROS Dox anticancer therapy

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Phase-separated bienzyme compartmentalization as artificial intracellular membraneless organelles for cell repair

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Science China Chemistry 2023年第3期66卷 845-852页

作者： Yanwen Zhang Shixin Wang Yuling Yan Xiaoxiao He Zefeng Wang Shaohong Zhou xiaohai yang Kemin Wang Jianbo Liu State Key Laboratory of Chemo/Biosensing and Chemometrics College of Chemistry and Chemical EngineeringKey Laboratory for Bio-Nanotechnology and Molecular Engineering of Hunan ProvinceHunan UniversityChangsha 410082China

Implanting artificial organelles in living cells is capable of correcting cellular dysfunctionalities for cell repair and biomedical applications. In this work, phase-separated bienzyme-loaded coacervate microdroplets are established as a model of artificial membraneless organelles in endothelial dysfunctional cells for the cascade enzymatic production of nitric oxide(NO) with a purpose of correcting cellular NO deficiency. We prepared the coacervate microdroplets via liquid-liquid phase separation of oppositely charged polyelectrolytes, in which glucose oxidase/horseradish peroxidase-mediated cascade reaction was compartmented. After the coacervate microdroplets were implanted in NO-deficient dysfunctional cells, the compartments maintained a phase-separated liquid droplet structure, which facilitated a significant enhancement of NO production in the dysfunctional cells. The recovery of NO production was further exploited to inhibit clot formation in blood plasma located in the cell suspension. This demonstrated a proof-of-concept design of artificial organelles in dysfunctional cells for cell repair and anticoagulation-related medical applications. Our results demonstrate an approach for the construction of coacervate droplets through phase separation for the generation of artificial membraneless organelles, which can be designed to provide an array of functionalities in living organisms that have the potential to be used in the field of cell engineering and medical therapy.

关键词： artificial membraneless organelles coacervate microdroplets nitric oxide liquid-liquid phase separation cell repair

Ultrasensitive monitoring of ribozyme cleavage product using molecular-beacon-ligation system

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Chinese Science Bulletin 2007年第5期52卷 603-607页

作者： MENG XiangXian TANG ZhiWen WANG KeMin TAN WeiHong yang xiaohai LI Jun GUO QiuPing State Key Laboratory of Chemo/Biosensing and Chemometrics Institute of Biological Technology College of Chemistry and Chemical Engineering Hunan University Changsha 410082 China

This paper reports a new approach to detect ribozyme cleavage product based on the molecular-beacon-ligation system. The molecular beacon, designed in such a way that one-half of its loop is complementary to ribozyme cleavage product, is used to monitor ligation process of RNA/DNA com-plex in a homogeneous solution and to convert directly cleavage product information into fluorescence signal. The method need not label ribozyme and ribozyme substrate, which is fast, simple and ultra-sensitive for detection of cleavage product. Detection limit of the assay is 0.05 nmol/L. The cleavage product of hammerhead ribozyme against hepatitis C virus RNA (HCV-RNA) was detected perfectly based on this assay. Owing to its ultrasensitivity, excellent specificity, convenience and fidelity, this method might hold out great promise in ribozyme reaction and ribozyme gene therapy.

关键词：核酶分裂产物核酸连接分子信标连接系统超灵敏监测

Novel protein detection method based on proximity- dependent polymerase reaction and aptamers

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Chinese Science Bulletin 2008年第2期53卷 204-208页

作者： yang xiaohai WANG Lei WANG KeMin TAN WeiHong TANG HongXing MENG XiangXian GUO QiuPing State Key Laboratory of Chemo/Biosensing and Chemometrics Biomedical Engineering Center College of Chemistry and Chemical Engineering and Key Laboratory for Bio-Nanotechnology and Molecular Engineering of Hunan Province Hunan University Changsha 410082 China

In recent years, specific detection of proteins is one of the hot issues about aptamers in proteomics. Here we reported a simple, sensitive and specific proximity-dependent protein assay with dual DNA aptamers. Thrombin was used as the model protein, and two aptamer probes with complementary se- quence at 3′-end were designed for the two distinct epitopes of the protein. Association of the two ap- tamers with thrombin resulted in stable hybrids due to the proximity of 3′-end, then polymerase reac- tion was induced. The amount of obtained dsDNA was indicated using the fluorescence dye Sybr Green I. The results showed that the initial velocity of polymerase reaction had a positive correlation with concentration of thrombin. The advantages of this dual-aptamer-based approach included simple and flexible design of aptamer probes, high selectivity and high sensitivity. The detection limit was 6.9 pmol/L.

关键词：核酸凝血酶聚合酶反应蛋白质检测

Monitoring p21 mRNA expression in living cell based on molecular beacon fluorescence increasing rate

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Chinese Science Bulletin 2008年第3期53卷 357-361页

作者： TANG HongXing yang xiaohai WANG KeMin TAN WeiHong LIU Bin HE LiFang WANG Wei State Key Laboratory of Chemo/Biosensing and Chemometrics Biomedical Engineering Center College of Chemistry and ChemicalEngineering and Key Laboratory for Bio-Nanotechnology and Molecular Engineering of Hunan Province Hunan UniversityChangsha 410082 China

Studying the expression level of mRNA in living cells will offer tremendous opportunities for ad-vancement in cell biology research,disease diagnostics,and drug *** this paper,a molecular beacon(MB)specific for the important tumor suppressor gene p21 has been designed and *** fluorescence signal was detected in real-time after the MB entered the cytoplasm of nasopharyn-geal carcinoma *** injecting the p21MB into nasopharyngeal carcinoma cell and p33-trans-fected nasopharyngeal carcinoma cell,the consistent increase of fluorescent signal intensity was de-tected in both cell lines,and maximum fluorescence intensity achieved in about 15 *** about 4 min following microinjection,the fluorescence increasing rate was significantly different between these two cell lines,which indicate the different p21 mRNA expression *** results obtained in the real-time detection were also validated by *** of the initial fluorescence increasing rate can effi-ciently reduce the side effect of enzyme and improve the accuracy in living cell mRNA detection.

关键词： p21 mRNA 鼻咽癌癌细腻荧光增强率基因表达