BIOSENSORS & BIOELECTRONICS
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2020
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IF 10.257
Kai Zhang,Zhenqiang Fan,Bo Yao,Tingting Zhang,Yuedi Ding,Sha Zhu,Minhao Xie
2019 novel coronavirus (2019-nCoV) with strong contagion in the crowd, has ravaged worldwide and severely impacts the human health and epidemic prevention system, by producing a series of significant stress reactions in the body to induce further cytokine storm. Transcription factors (TFs) served as essential DNA binding proteins play an integral role in regulating cytokine storm, and the detection of it in the human coronavirus environment provides especially valuable approaches to diagnosis and treatment of 2019-nCoV and development of antiviral drugs. In this work, an entropy-driven electrochemiluminescence (ECL) biosensor was constructed for ultra-sensitive bioassay of NF-κB p50. The strategy primarily capitalizing the splendid double-stranded DNA (dsDNA) binding properties of transcription factors, employing GOAu-Ru composite material as ECL emitter, utilizing entropy-driven reactions for signal amplification method, offered a repeatable proposal for TFs detection. In the absence of TFs, the released DNA1 further went in the entropy-driven reaction, contributing to an “ECL off” state. However, in the presence of TFs, the dsDNA avoided being digested, which blocked DNA1 for participating in the entropy-driven reaction, and the system exhibited an “ECL on” state. Most importantly, the ECL bioanalytical method denoted broad application prospects for NF-κB p50 detection with a lower detection limit (9.1 pM).
研究领域:
分析化学
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生物化学
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分子生物学
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病毒学
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医学诊断
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DOI
PubMed
高影响因子
MICROCHEMICAL JOURNAL
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2025
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IF 5.1
Jiaying Wang,Yu Tian,Kai Zhang,Jiantong Jiao,Yi Zhao
NF-κB p50 is a critical transcription factor that plays an essential role in regulating immune responses, inflammatory pathways, and tumor progression. Its aberrant expression is closely linked to various pathological conditions, making it a valuable biomarker for early disease diagnosis and therapeutic monitoring. In this work, we developed a highly sensitive and reusable electrochemical biosensor capable of detecting NF-κB p50 at ultralow concentrations. The biosensor integrates a pH-responsive regeneration mechanism and a multi-stage signal amplification cascade involving Exonuclease III (Exo III)-assisted recycling, CRISPR/Cas12a-mediated trans -cleavage, and DNA tetrahedron nanostructure-functionalized electrodes. This synergistic design significantly enhances detection sensitivity and stability. The platform offers a broad dynamic detection range from 5 aM to 100 fM with a remarkably low detection limit of 2.38 aM. More importantly, the biosensor demonstrates excellent reusability through efficient regeneration under alkaline conditions (pH 10.0), with minimal loss in signal output. Validation in tenfold-diluted human serum samples yielded high recovery rates (99.61 %–106.23 %) and low relative standard deviations, confirming its practical applicability. This study not only presents a robust approach for ultrasensitive protein detection but also introduces a generalizable strategy for the regenerative electrochemical analysis of protein-based biological macromolecules in clinical diagnostics.
研究领域:
生物医学工程
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分子诊断学
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纳米技术
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DOI
高影响因子
MICROCHEMICAL JOURNAL
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2025
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IF 5.1
Dianbo Yu,WenZhao Zhang,Jiahou Xu,Yuanxun Gong,Jiayi Zhang,Jihua Wei,Kai Zhang,Xianjiu Liao
Nuclear factor kappa B (NF-κB) p50 is a critical transcription factor that regulates immune responses, inflammation, and cell proliferation. Precise detection of NF-κB p50 at ultralow concentrations is of paramount importance for early disease diagnosis and mechanistic studies. Here, we report an electrochemical biosensor integrating double-crossover DNA tiles and entropy-driven strand displacement, amplified by Exonuclease III (Exo III) digestion. By designing a self-assembled DNA tile scaffold on a gold electrode and employing a DNA tetrahedron architecture, the sensor achieves a highly ordered surface configuration, minimizing background noise and enhancing specificity. In the absence of NF-κB p50, Exo III selectively digests a double-stranded DNA substrate, releasing single-stranded triggers that undergo an entropy-driven reaction to form a Y-shaped scaffold on the electrode. This assembly effectively hinders redox probe access, lowering the differential pulse voltammetry (DPV) signal. Conversely, NF-κB p50 binding protects the DNA substrate from digestion, preserving a higher current output. Under optimized conditions—30 nM DNA tiles, 2 U μL −1 Exo III, 30 min cleavage, and 100 nM DNA3/DNA4/DNA5—the biosensor exhibits an ultralow detection limit of 2.65 aM and excellent specificity, as confirmed by negligible cross-reactivity with other proteins. Furthermore, the method shows robust anti-interference performance in complex matrices (e.g., diluted cell lysates). These findings underscore the potential of this DNA tile-based biosensor for clinical diagnostics and fundamental research on NF-κB p50–mediated pathways.
研究领域:
生物传感器
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分子诊断
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生物电子学
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核酸纳米技术
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转录因子
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DOI
高影响因子
ACS Applied Materials & Interfaces
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2020
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IF 8.758
Zhenqiang Fan,Zongqiong Lin,Zepeng Wang,Jianfeng Wang,Minhao Xie,Jianfeng Zhao,Kai Zhang,Wei Huang
In this work, we fabricated a dual-wavelength electrochemiluminescence ratiometric biosensor based on electrochemiluminescent resonance energy transfer (ECL-RET). In this biosensor, Au nanoparticle-loaded graphitic phase carbon nitride (Au-g-C3N4) as a donor and Au-modified dimethylthiodiaminoterephthalate (TAT) analogue (Au@TAT) as an acceptor were investigated for the first time. Besides, tetrahedron DNA probe was immobilized onto Au-g-C3N4 to improve the binding efficiency of the transcription factor and ECL ratiometric changes on the basis of the ratio of ECL intensities at 595 and 460 nm, which were obtained through the formation of a sandwich structure of DNA probe–antigen–antibody. Our biosensor achieved the assay of NF-κB p50 with a detection limit of 5.8 pM as well as high stability and specificity.
研究领域:
分析化学
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生物传感器
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电化学
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纳米技术
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分子生物学
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DOI
PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
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2022
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IF 12.545
Feng Qiu,Xiaoyan Gan,Jianglong Yao,Bingying Jiang,Ruo Yuan,Yun Xiang
Transcription factors (TFs) are key substances in regulating the transcription, replication and expression of genes, and the detection of TFs can provide valuable information to diagnose a variety of diseases. By integrating hybridization chain reaction (HCR)-activated Cas12a enzyme with bio-responsive DNA hydrogels, we propose a dual amplification and label-free homogeneous electrochemical detection method to realize sensitive nuclear factor-kappa B p50 (NF-κB p50) detection. The presence of the target molecules protects the DNA duplex probes from digesting by exonuclease III and initiates HCR to generate long double stranded DNAs that can activate the activity of RNA-guided Cas12a enzymes. The single-stranded region of the DNA linkers that crosslink the DNA hydrogels can be cleaved by the activated Cas12a to release a large number of electroactive substances embedded in the gels, which exhibit highly enhanced electrochemical signals for detecting target molecules at the detection limit of 54.1 fM. In addition, the successful interrogation of NF-κB p50 spiked into lysate of HeLa cells by such method is also verified. The established method thus shows new opportunities for sensitive and convenient monitoring of other transcription factors and biomarkers.
研究领域:
生物化学
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分子生物学
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医学诊断
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产品关联
DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
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2018
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IF 5.667
Bingzhi Li,Yue Chen,Jing Wang,Qiaoyun Lu,Wanying Zhu,Lei Xu,Xin Shen,Jieping Luo,Chunhong Zhu,Xiaoxu Li,Junli Hong,Xuemin Zhou
Transcription factors (TFs) regulate information flow from gene to protein, and they are recognized as key indicators to reflect cellular processes. Facile monitoring of TFs may aid diagnostics and treatment, but we still lack of techniques for highly efficient detection of them. In this research, a hybrid material consisting of DNA-silver nanoclusters (DNA-AgNCs) and polypyrrole nanoparticles (PPyNPs) was built for TFs detection. The designed DNA-AgNCs have a hairpin-shaped nucleic acid architecture with a double-stranded stem for recognizing TFs and a single-stranded loop for interacting with PPyNPs. In the absence of TFs, DNA-AgNCs are absorbed to PPyNPs, resulting in fluorescent quenching. While in the presence of TFs, the binding between TFs and the DNA-AgNCs caused desorption of DNA-AgNCs via steric hindrance mechanism. Accordingly, the increase of fluorescence derived from desorption is used for quantifications. Derived from low non-specific protein absorption features of PPyNPs, detection limit of 70 pM for NF-κB p50 and 110 pM for p53 were obtained. Then, by absorbing two kinds of DNA-AgNCs to PPyNPs, label-free multiplexed detection of TFs was first realized. Additionally, we suggested that this platform can be developed for drug screening by evaluating inhibitory effect of a pair of optical isomers towards TFs.
研究领域:
纳米技术
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生物医学工程
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分析化学
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分子生物学
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DOI
高影响因子
BIOCHEMICAL PHARMACOLOGY
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2026
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IF 6.5
Tianyue Guan,Audrey Dela,Yichen Jin,Juqin Song,Tingyan Wang,Yu Zhu,Jingquan Dong,Panpan Zhao
Sepsis-induced acute lung injury (ALI) remains a critical factor contributing to mortality in intensive care units, and effective therapeutic strategies are still lacking. The present research explores the defensive impacts and associated mechanisms of Tubuloside A (TA), an active phenanthrenoid glycoside extracted from Cistanche deserticola, in sepsis-induced ALI. Utilizing both a cecal ligation and puncture (CLP) mouse model and LPS-stimulated MLE-12 alveolar epithelial cells, we found that TA markedly attenuated lung injury, as indicated by improved histopathological features, reduced pulmonary edema, and maintenance of alveolar structure. Mechanistically, TA provided multifaceted protection by concurrently inhibiting oxidative stress, ferroptosis, and inflammatory signaling—critical drivers of sepsis-induced ALI. Treatment with TA diminished lipid peroxidation, restored GPX4 expression, and inhibited pro-inflammatory cytokines (TNF-α, IL-6, IL-1β). Network pharmacology identified NFKB1 as a therapeutic target of TA against ALI. Molecular docking predicted and SPR analysis verified that TA binds to NF-κB p50. Notably, functional validation via NF-κB p50 overexpression abolished TA-mediated protection and re-sensitized cells to ferroptosis, confirming NF-κB p50 as a central regulatory node. Furthermore, TA activated Nrf2 signaling in an NF-κBp50–dependent manner, as demonstrated by nuclear translocation assays and rescue experiments. Calcein AM imaging and iron chelation studies confirmed that TA’s anti-ferroptotic effect requires functional NF-κB/Nrf2/GPX4 crosstalk. Collectively, our findings reveal that TA ameliorates sepsis-ALI by targeting the NF-κBp50–Nrf2/GPX4 axis to coordinately inhibit inflammation, oxidative stress, and ferroptosis, highlighting its potential as a multi-target phytotherapeutic agent for acute respiratory disorders
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高影响因子
ANALYTICAL CHEMISTRY
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2017
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IF 6.32
Bingzhi Li,Lei Xu,Yue Chen,Wanying Zhu,Xin Shen,Chunhong Zhu,Jieping Luo,Xiaoxu Li,Junli Hong,Xuemin Zhou
Transcription factors (TFs) regulate gene expression by binding to regulatory regions, and their dysregulation is involved in numerous diseases. Thus, they are therapeutic targets and potential diagnostic markers. However, widely used methods for TFs detection are either cumbersome or costly. Herein, we first applied DNA-Ag nanoclusters molecular beacons (AgMBs) in TFs analysis and designed an assay based on the switchable fluorescence of AgMBs. In the absence of TFs, a single-stranded DNA functioned as a reporter is released from a double-stranded DNA probe (referred as dsTFs probe) under exonuclease III (Exo III) digestion. Then, the reporter triggers downstream Exo III-assisted signal amplification by continuously consuming the guanine-rich enhancer sequences in AgMBs, resulting in significant fluorescent decrease eventually. Conversely, the presence of TFs protects the dsTFs probe from digestion and blocks the downstream reaction to keep a highly fluorescent state. To testify this rationale, we utilized nuclear factor-kappa B p50 (NF-κB p50) as a model TFs. Owing to the amplification strategy, this method exhibited high sensitivity toward NF-κB p50 with a limit of detection of 10 pM, and a broad linear range from 30 pM to 1.5 nM. Furthermore, this method could detect multiple TFs in human colon cancer DLD-1 cells and reflect the variation in their cellular levels after stimulation. Finally, by conducting an inhibition assay we revealed the potential of this method for screening TFs-targeted drugs and calculating the IC50 of corresponding inhibitors.
研究领域:
生物化学
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分子生物学
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生物技术
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癌症研究
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药物发现
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产品关联
DOI
PubMed
高影响因子
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