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BIOSENSORS & BIOELECTRONICS · 2016 · IF 7.476

Ultrasensitive non enzymatic multiple immunosensor for tumor markers detection by coupling DNA hybridization chain reaction with intercalated molecules

Jinjin Guo,Junchun Wang,Junjun Zhang,Wenjuan Zhang,Yuzhong Zhang

In this study, we tried coupling the small signal molecules that could intercalate into DNA double helix with hybridization chain reaction (HCR) technique to fabricate a multiple immunosensor. Doxorubicin hydrochloride (DXH) and methylene blue (MB) were used as signal molecules and alpha-fetoprotein (AFP) and carcinoembryonic antigen (CEA) were selected as model biomarkers. The immunosensor mainly consists of three parts as follows: First, two different primary antibodies (Ab 1 ) immobilized on the surface of gold nanoparticles (Au NPs); Second, secondary antibodies (Ab 2 ) conjugated with DNA primer ; Third, long DNA concatemers from HCR were used as a carrier to intercalate amounts of signal molecules (DXH or MB). A sandwich immunocomplex was formed among primary antibodies, target biomarkers and secondary antibodies conjugated with DNA primer via specific recognition reaction. Afterwards, DNA concatemers intercalating amounts of DXH or MB were linked to DNA primer via DNA hybridization. Square wave voltammetry (SWV) was employed to record the response signals from electroactive molecules DXH and MB, and two distinguishable signals were obtained, which peak potentials were at about −0.30   V (corresponding to MB) and −0.70   V (corresponding to DXH, both vs SCE), respectively. The signal intensities of MB and DXH were linearly related to the logarithm of biomarkers concentration in the range of 0.05   pg   mL −1 –25   ng   mL −1 , and the limit of detection were 0.03   pg   mL −1 for CEA and 0.02   pg   mL −1 for AFP (at S/N=3), respectively. Furthermore, the immunosensor exhibited a sensitive electrochemical response to biomarkers in human serum samples and the results obtained were in accordance with reference method, indicating the immunosensor can be applied to real sample analysis in clinic diagnosis.

CAS / MDL: 25952-53-8 6066-82-6
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MOLECULES · 2017 ·Vol. 22 ·期 1 ·171 · IF 2.861

In Vitro DNA-Binding, Anti-Oxidant and Anticancer Activity of Indole-2-Carboxylic Acid Dinuclear Copper(II) Complexes

Xiangcong Wang,Maocai Yan,Qibao Wang,Huannan Wang,Zhengyang Wang,Jiayi Zhao,Jing Li,Zhen Zhang

Indole-2-carboxylic acid copper complex (ICA-Cu) was successfully prepared and characterized through elemental analysis, IR, UV-Vis,1H-NMR, TG analysis, and molar conductance, and its molecular formula was [Cu2(C9H6O2N)4(H2O)2]·2H2O. The binding ability of ICA-Cu to calf thymus DNA (CT-DNA) was examined by fluorescence spectrometry and the viscosity method. The results indicated that, upon the addition of increasing amounts of CT-DNA, the excitation and emission intensity of ICA-Cu decreased obviously and the excitation spectra shifted towards a long wavelength. ICA-Cu could displace ethidium bromide (EB) from the EB-DNA system, making the fluorescence intensity of the EB-DNA system decrease sharply; the quenching constantKSVvalue was 3.99 × 104M−1. The emission intensity of the ICA-Cu-DNA system was nearly constant, along with the addition of Na+in a series of concentrations. The fluorescence of the complex could be protected after the complex interacted with DNA. A viscosity measurement further supported the result that the ICA-Cu complex may interact with DNA in an intercalative binding mode. The antioxidant activities of ICA-Cu were evaluated by a 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay, a hydroxyl radical (OH) scavenging assay, and a 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid (ABTS) assay. The ICA-Cu exhibited the highest inhibitory effects on the ABTS radical (94% inhibition at 60 µM), followed by OH and DPPH radicals (the degrees of inhibition being 71% and 56%, respectively). The in vitro cytotoxicity activity of ICA-Cu against two human breast cancer cell lines, MDA-MB-231 and MCF-7, was investigated by 3-[4,5-dimethyltiazol2-yl]-2.5-diphenyl-tetrazolium bromide (MTT) assay and cellular morphological analysis. The results showed that, upon increasing the concentration of ICA-Cu, an increase was observed in growth-inhibitory activity and the inhibition percentage were greater than 90% at 20 µM in both cell lines. Also, cellular morphological changes in the two cell lines agreed with the cytotoxicity results.Keywords:DNA-binding;antioxidant;anticancer;indole-2-carboxylicacid;copper(II) complexes

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Analytical Methods · 2017 ·Vol. 9 ·期 7 ·1105-1111 · IF 1.9

Electrochemical detection of DNA damage induced by clenbuterol at a reduced graphene oxide-Nafion modified glassy carbon electrode

Xiaoyun Lin,Yongnian Ni,Xueying Pei,Serge Kokot

An electrochemical biosensor capable of direct detection of DNA damage induced by clenbuterol (CLB) has been developed. A glassy carbon electrode (GCE) was modified using reduced graphene oxide-Nafion and dsDNA to produce a dsDNA/RGO-Nafion/GCE sensor. The modified electrode was characterized by electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV), and the results at each stage of the electrode construction were interpreted. They indicated that the electrochemical oxidation peak currents of guanine and adenine at the electrode (dsDNA/RGO-Nafion/GCE) significantly increased as compared with those at the untreated electrode (dsDNA/GCE). Based on this, the sensor was used to measure the change of the oxidation peak currents for guanine and adenine between the intact and damaged dsDNA in the sensor films, and the DNA damage was successfully detected. Furthermore, the plot of peak current decline for the guanine and adenine versus the concentration of CLB was linear in the range of 5.0 × 10−7 to 4.0 × 10−6 mol L−1, with a limit of detection (LOD) of 3.2 × 10−7 mol L−1, which provided a good method to determine CLB indirectly.

CAS / MDL: 21898-19-1 31175-20-9
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CELLULOSE · 2024 · IF 4.9

Eco-friendly L-cysteine and DNA co-anchored Ag NPs on cotton fabrics for durable wearable joule heaters

Xiang Shuangfei,Cai Dongrong,Ru Hongfu,Ma Yuchen,Xiao Yuanxiang,Zhao Shujun,Fu Feiya,Liu Xiangdong

The rational design of Ag NP decorated cotton fabrics with durable Joule heating performance has attracted great interest. However, the combination of Ag NPs and cotton fabrics is so weak that it hinders its long-term stability in service. Here, an in situ dual-anchoring strategy is employed to enhance the immobilization of Ag NPs by L-Cysteine and DNA on cotton fabrics. Specifically, DNA plays a role as template for immobilizing Ag NPs, while L-Cysteine acts as a binder between cotton fabrics and DNA. Under the synergistic effect of L-Cysteine and DNA, Ag NPs are firmly anchored on cotton fabrics. The obtained cotton fabric based wearable Joule heaters exhibit low electrical resistance (24 Ω/sq) and long-term stability. Furthermore, the cotton fabric based wearable Joule heaters can be heated up to 60.3℃ at a low driving voltage of 4.5 V, and maintain superior Joule heating performance even after 1000 times of bending fatigue tests. Thus, this work holds great promise for a durable wearable Joule heater. Graphical abstract

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Analytical Methods · 2024 · IF 2.7

Highly sensitive electrochemical biosensor for MUC1 detection based on a DNA functionalized CdTe quantum dots as signal enhancers

Bao-Kang Jin,Jiajia Wang,Chun Kan

In this paper, a proportional electrochemical biosensor based on cadmium telluride/ polypyrrole (CdTe/PPy) nanocomposite is developed for the detection of MUC1 with high selectivity and sensitivity. The result indicates that CdTe/PPy nanocomposite is modified on the surface of glassy carbon electrode (GCE), which supplies a large surface area for immobilizing cap-DNA, ensuring its high selectivity and sensitivity. Whereafter, CdTe linked sig-DNA (MUC1 aptamer) is introduced, allowing MUC1 aptamer to hybridize with cap- DNA. CdTe as signal amplification elements in order to generate the differential pulse voltammetry (DPV) signal. Conceivably, the target MUC1 detection was based upon the current signal change due to the quantities change of the signal amplification elements CdTe. In the presence of MUC1, the MUC1 aptamer specifically binds to MUC1, resulting in the release of CdTe-sig-DNA from the electrode surface and the decrease of peak current. Under the optimized experimental conditions, the electrochemical biosensor present a highly selective, sensitive, stability and reproducibility for the MUC1 from the range of 0.1 nM to 100 nM with detection limit of 0.05 nM (S/N = 3). Therefore, the electrochemical biosensor has a potential application in medical disease diagnosis application detection.

✓ 已验证 产品关联 DOI PubMed 高影响因子
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MICROCHEMICAL JOURNAL · 2024 · IF 4.9

Fluorescent sensing platform based on two-dimensional layered double hydroxides and exonuclease I for DNA and microRNA detection

Yuanyi Wu,Yan Yang,Shihao Yang,Yang Ju,Yang Jiang,Yumei Xiao,Jialin He,Tian Liu,Leyao Tang,Kaiyong Wang,Yi Yang

Interfacial and interlayer interactions between layered double hydroxides (LDHs) and nucleic acids offer LDHs great potential to absorb and detect nucleic acids. Herein, we demonstrate a simple, rapid, and versatile fluorescent sensing platform based on two-dimensional LDHs and exonuclease I (Exo I) for the selective analysis of DNA and microRNA. ZnNiAl-LDHs were utilized as quenchers for FAM labelled on single-stranded DNA (ssDNA) probes via dynamic quenching with high fluorescence quenching ability. The quenching mechanism was explored by means of spectral analysis, characterization experiments, DNA adsorption and desorption studies, zeta potential analysis, and molecular dynamics simulations. The driving forces for the adsorption of LDHs toward fluorophore-labelled nucleic acids were confirmed to be electrostatic interaction, van der Waals force, and anion exchange for the first time. In order to distinguish the formed target/probe duplexes from FAM-ssDNA probes, Exo I was introduced to specifically hydrolyze FAM-ssDNA probes. The fabricated fluorescent sensing platform for nucleic acids detection shows a low detection limit of 0.08 nM. Finally, this sensor was effectively utilized for the detection of DNA and microRNA in actual biological and environmental samples, achieving recoveries ranging from 96.4 % to 105 %, indicating high accuracy. This work proposed a promising sensing platform for sequence-specific nucleic acids detection, and provides a new perspective on two-dimensional LDHs as structural supports and biomolecular reservoirs for biomedical applications.

CAS / MDL: 1185-53-1
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Cell Death & Disease · 2025 ·Vol. 16 ·期 1 ·1-14 · IF 9.6

Mcm5 mutation leads to silencing of Stat1-bcl2 which accelerating apoptosis of immature T lymphocytes with DNA damage

Liu Min,Li Yuanyuan,Deng Zhilin,Zhang Ke,Huang Shuying,Xia Jiamin,Feng Yi,Liang Yundan,Sun Chengfu,Liu Xindong,Li Shurong,Su Bingyin,dong Yong,Huang Sizhou

Mutation in genes involved in DNA replication continuously disrupt DNA replication and give rise to genomic instability, a critical driver of oncogenesis. To prevent leukemia, immature T lymphocytes with genomic instability often undergo rapid cell death during development. However, the mechanism by which immature T lymphocytes undergo rapid cell death upon genomic instability has been enigmatic. Here we show that zebrafish mcm5 mutation leads to DNA damage in immature T lymphocytes and the immature T cells sensitively undergo rapid cell death. Detailed analyses demonstrated that the immature T lymphocytes undergo rapid apoptosis via upregulation of tp53 and downregulation of bcl2 transcription in mcm5 mutants. Mechanistically, Mcm5 directly binds to Stat1a and facilitates its phosphorylation to enhance bcl2a expression under the conditions of DNA replication stress. However, in mcm5 mutants, the absence of the Mcm5-Stat1 complex decreases Stat1 phosphorylation and subsequent bcl2a transcription, accelerating apoptosis of immature T lymphocytes with genomic instability. Furthermore, our study shows that the role of Mcm5 in T-cell development is conserved in mice. In conclusion, our work identifies a role of Mcm5 in regulating T cell development via Stat1-Bcl2 cascade besides its role in DNA replication, providing a kind of mechanism by which immature T cells with gene mutation-induced DNA damage are rapidly cleared during T lymphocyte development.

CAS / MDL: 119-90-4
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JOURNAL OF CONTROLLED RELEASE · 2024 · IF 10.5

Nuclear-targeted chimeric peptide nanorods to amplify innate anti-tumor immunity through localized DNA damage and STING activation

Yeyang Wu,Yanmei Li,Ni Yan,Jiaqi Huang,Xinyu Li,Keyan Zhang,Zhenming Lu,Ziwen Qiu,Hong Cheng

Stimulator of the interferon genes (STING) pathway is appealing but challenging to potentiate the innate anti-tumor immunity. In this work, nuclear-targeted chimeric peptide nanorods (designated as PFPD) are constructed to amplify innate immunity through localized DNA damage and STING activation. Among which, the chimeric peptide (PpIX-FFVLKPKKKRKV) is fabricated with photosensitizer and nucleus targeting peptide sequence, which can self-assemble into nanorods and load STING agonist of DMXAA. The uniform nanosize distribution and good stability of PFPD improve the sequential targeting delivery of drugs towards tumor cells and nuclei. Under light irradiation, PFPD produce a large amount of reactive oxygen species (ROS) to destroy nuclear DNA in situ , and the released cytosolic DNA fragment will efficiently activate innate anti-tumor immunity in combination with STING agonist. In vitro and in vivo results indicate the superior ability of PFPD to activate natural killer cells and T cells, thus efficiently eradicating lung metastatic tumor without inducing unwanted side effects. This work provides a sophisticated strategy for localized activation of innate immunity for systemic tumor treatment, which may inspire the rational design of nanomedicine for tumor precision therapy.

CAS / MDL: 553-12-8 6485-79-6
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JOURNAL OF ELECTROANALYTICAL CHEMISTRY · 2018 · IF 3.235

A simple and universal electrochemical assay for sensitive detection of DNA methylation, methyltransferase activity and screening of inhibitors

Jing Bao,Xintong Geng,Changjun Hou,Yanan Zhao,Danqun Huo,You Wang,Zhaonan Wang,Yan Zeng,Mei Yang,Huanbao Fa

Many studies have confirmed that DNA methylation is highly correlated with the occurrence and development of various diseases including cancers. In this work, we developed a simple, sensitive, selective, and universal electrochemical biosensor for detection of DNA methylation and assay of DNA methyltransferase (MTase) activity using M.SssI MTase as an example. The thiolated single-stranded DNA S1 was self-assembled on the surface of gold nanoparticles deposition modified glassy carbon electrode via Au S bonding, then hybridization between the DNA S1 and its complementary DNA S2 formed a double-stranded target sequence for both M.SssI MTase and restriction endonuclease Hpa II. HpaII could not cleave the target sequence after it was methylated by M.SssI MTase, while the sequence without methylation could be cleaved. Here, we used methylene blue (MB) as electrochemical indicator. The electrochemical signal of MB increased linearly with increasing M.SssI MTase concentration from 0.5 to 25 U/mL and from 25 to 400 U/mL with a detection limit of 0.04 U/mL. Moreover, screening of M.SssI MTase inhibitors 5-azacytidine (5-Aza) and 5-Aza-2′-deoxycytidine (5-Aza-dC) were successfully investigated using the fabricated electrochemical biosensor and showed that the two classic drugs could both inhibit the M.SssI MTase activity with the IC 50 of 2.8 μM and 0.37 μM, respectively, indicating potential application in discovery of new anticancer drugs.

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ACS Applied Nano Materials · 2025 · IF 5.5

DNA-Templated Ag Nanocluster/CdSe@ZnS Quantum Dot-Based Platform for Colorimetric and Fluorescence Detection of Organophosphorus Pesticide

Xiaoxia Kang,Jige Yuan,Xijuan Gu,Xiaoyan Li,Tao Wen,Miao He,Yue He,Xiaoxiao Yang,Yuechen Li,Conglin Guo,Haiwei Ji,Yuling Qin,Li Wu

The agriculture and environmental industries have employed organophosphorus pesticides (OPs) chemicals as insecticides extensively in previous decades, leading to persistent life-threatening issues. Therefore, developing a platform for the efficient identification of OPs is significant for environmental protection and human health. In this study, a strategy for the colorimetric and fluorescence detection of OPs by combining quantum dots (QDs) with DNA-templated silver nanoclusters (DNA-AgNCs) was proposed. Technically, the fluorescence of DNA-AgNCs will be enhanced once guanine (G)-rich sequences get close to the DNA-AgNCs to form hybrid complexes DNA-AgNCs/G, while quenching of fluorescence can be attributed to the hydrolysate (thiocholine) of acetylthiocholine (ATch) that was caused by acetylcholinesterase (AchE). Based on the fact that OPs can irreversibly inhibit the catalytic activity of AchE and slow down the fluorescence quenching of DNA-AgNCs/G, thus, OPs detection can be achieved by evaluating the fluorescence intensity of DNA-AgNCs/G. Additionally, a dual-mode detection system (colorimetric and fluorescent) is achieved through an ion-exchange reaction between QDs and DNA-AgNCs/G. The concentration of OPs is quantified by monitoring fluorescence changes in the QDs and poly(9,9-dioctylfluorene) (PFO) polymer dots (Pdots) system, with PFO Pdots serving as a stable internal reference. Using trichlorfon as a model, the limit of detection (LOD) was 2.55 pg/mL, while an excellent linear relationship with the trichlorfon concentrations in the range of 5 × 100–5 × 105 pg/mL was obtained. Additionally, visual detection (as low as 1.06 pg/mL) can be realized readily, taking advantage of the ion-exchange reaction between DNA-AgNCs and QDs.

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ANALYTICAL CHEMISTRY · 2024 · IF 6.7

DNA Conformation-Regulated Hemin Switch for Lab-on-Chip Chemiluminescent Detection of an Antibody Secreted from Hybridoma Cells

Hang Ao,Wencheng Xiao,Wenrui Hu,Jie Wu,Huangxian Ju

This work designed a DNA conformation-regulated hemin switch for rapid chemiluminescent (CL) detection of a monoclonal antibodies. This switch was performed with an affinity probe and an inhibition probe, which were conveniently prepared by hybridizing hemin-labeled DNA1 with KHL peptide-labeled DNA2 and binding biotin-labeled DNA3 to streptavidin, respectively. In the absence of the target antibody, streptavidin-DNA3 could hybridize with hemin-DNA1/KHL-DNA2 to release KHL-DNA2, which led to the loss of hemin activity due to the affinity hindrance of streptavidin-DNA3. After the KHL peptide was recognized by the target antibody, the strand replacement hybridization could be inhibited by the bound antibody, which retained the high catalytic activity of hemin overhung on the antibody-bound affinity probe for a CL reaction, leading to a “signal-on” process for CL antibody detection. Using a KHL-specific antibody, anti-proprotein convertase subtilisin/kexin type 9 antibody (PCSK9-Ab), as a target model and common L012–1,2,4-triazole–H2O2 CL system, the designed switch showed a detection range of 10 ng mL–1 to 1 μg mL–1 with a detection limit of 4.16 ng mL–1 (56.2 pM) and a short analytical time of 6.5 min. The proposed quick method could simply be used for lab-on-chip CL detection of PCSK9-Ab in situ-secreted from PCSK9-6E3 hybridoma cells, which showed an accuracy of 90.2% compared with the statistical results from general fluorescence imaging, providing a potential technique for screening specific hybridoma cells.

CAS / MDL: 9013-20-1
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Acta Biomaterialia · 2024 · IF 9.4

A DNA-inspired injectable adhesive hydrogel with dual nitric oxide donors to promote angiogenesis for enhanced wound healing

Yongsheng Zheng,Dong Yang,Botao Gao,Shuai Huang,Yubo Tang,Qingde Wa,Yong Dong,Shan Yu,Jun Huang,Sheng Huang

Chronic diabetic wounds are a severe complication of diabetes, often leading to high treatment costs and high amputation rates. Numerous studies have revealed that nitric oxide (NO) therapy is a promising option because it favours wound revascularization. Here, base-paired injectable adhesive hydrogels (CAT) were prepared using adenine- and thymine-modified chitosan (CSA and CST). By further introducing S-nitrosoglutathione (GSNO) and binary L-arginine (bArg), we obtained a NO sustained-release hydrogel (CAT/bArg/GSON) that was more suitable for the treatment of chronic wounds. The results showed that the expression of HIF-1α and VEGF was upregulated in the CAT/bArg/GSON group, and improved blood vessel regeneration was observed, indicating an important role of NO. In addition, the research findings revealed that following treatment with the CAT/bArg/GSON hydrogel, the viability of Staphylococcus aureus and Escherichia coli decreased to 14 ± 2% and 6 ± 1%, respectively. Moreover, the wound microenvironment was improved, as evidenced by a 60 ± 1% clearance of DPPH. In particular, histological examination and immunohistochemical staining results showed that wounds treated with CAT/bArg/GSNO exhibited denser neovascularization, faster epithelial tissue regeneration, and thicker collagen deposition. Overall, this study proposes an effective strategy to prepare injectable hydrogel dressings with dual NO donors. The functionality of CAT/bArg/GSON has been thoroughly demonstrated in research on chronic wound vascular regeneration, indicating that CAT/bArg/GSON could be a potential option for promoting chronic wound healing. Statement of significance This article prepares a chitosan hydrogel utilizing the principle of complementary base pairing, which offers several advantages, including good adhesion, biocompatibility, and flow properties, making it a good material for wound dressings. Loaded GSNO and bArg can steadily release NO and L-arginine through the degradation of the gel. Then, the released L-arginine not only possesses antioxidant properties but can also continue to generate a small amount of NO under the action of NOS. This design achieves a sustained and stable supply of NO at the wound site, maximizing the angiogenesis-promoting and antibacterial effects of NO. More neovascularization and abundant collagen were observed in the regenerated tissues. This study provides an effective repair hydrogel material for diabetic wound.

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