MICROCHEMICAL JOURNAL
·
2025
·
IF 5.1
Yan Yang,Wenxin Mai,Shuojie Wang,Guixiang Teng,Shiyu Wei,Meiling Shi,Yongqiang Tian
Ochratoxin A (OTA) is a widespread mycotoxin in food chains, yet conventional aptamer-based assays remain challenging because of the background signals and matrix interference. Here, we introduced a target-triggered reporter synthesis mechanism in which OTA binding unlocks a TdT- Nt.AlwI enzymatic cycle that generates polyT strands exclusively in the presence of the target. These generated polyT products interact specifically with polyA-recognition motifs on DNA-templated silver nanoclusters (DNA-AgNCs), enabling fluorescence transduction for OTA detection. This “zero-to-one” generative logic gate ensures that signal molecules are produced exclusively after recognition, intrinsically suppressing blank background. Moreover, the enzymatic cycle allows a single OTA molecule to trigger the generation of multiple polyT strands, imparting signal amplification without external labels or immobilization. Furthermore, integrating target-initiated nucleic acid synthesis with orthogonal optical transduction enables the construction of a label-free assay that intrinsically decreases background noise from signal generation, thereby ensuring high specificity, low interference, and quantitative reliability for target detection. The assay shows a linear response from 10 pM to 50 nM, achieves a detection limit of 0.097 ng mL −1 , and yields 86.4–94.1 % recoveries with<8.1 % RSD in spiked soybean extracts. By suppressing baseline noise at the mechanism level and separating amplification from readout, this platform enables sensitive, low false-positive OTA quantification and is readily adaptable to other small molecules by exchanging the aptamer, advancing portable food-safety screening.
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Journal of Pharmaceutical Analysis
·
2025
·
IF 8.9
Shiquan Zheng,Jiale Ke,Hanren Chen,Huaze Shao,Fengxin Zheng,Runhui Zhang,Zean Zhao,Jianxin Pang,Lihong Liu
The lack of a cell-based screening method limits urate-lowering drug development. A novel method combining aptamer sensor array (ASA), exonuclease III (Exo III)- powered 3D DNA walker (DW), and linear discriminant analysis (LDA) was developed for detecting uric acid (UA) in cell lysates, referred to as ASA–Exo III-DW–LDA. Three aptamers (Apts) with different affinities for UA and its structurally similar compound, xanthine (Xan), were used to design the ASA. The combination of ASA and Exo III-DW enabled the detection of UA at the picomolar level, whereas LDA was employed to differentiate UA signals from the mixed signals of UA and Xan. Significantly, Pearson correlation analysis revealed a strong correlation between our method and the 14 C radioactive labeling method for urate anion exchanger 1 (URAT1) inhibitors, with r = 0.9880 for lesinurad and r = 0.9777 for benzbromarone. Using our method, kaempferol was identified as a promising hit compound for inhibiting the URAT1, because of its low half-maximal inhibitory concentration (IC 50 ) (18.96 μM) low toxicity in mouse renal tubular epithelial cells (mTECs), and significant urate-lowering effect in hyperuricemic mice at 5 mg/kg. Overall, this method is sensitive, cost-effective and safe, offering a novel strategy for routine urate-lowering drug screening in standard laboratories.
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ANALYTICAL CHEMISTRY
·
2025
·
IF 6.7
Yunshan Zhang,Qianglong Tan,Fang Yang,Tuo Huang,Siyu Yu,Jing Ye,Jianxian Zeng,Xianzhong Feng,Diming Zhang
DNA walkers, as structurally and functionally programmable signal amplification tools, exhibit great potential for application in the field of biosensing. Traditional DNA walkers often rely on enzymes for operation, posing compatibility challenges, while the handful of existing enzyme-free DNA walkers demonstrate limited performance. To address this, we innovatively developed an efficient enzyme-free 3D DNA walker with dual capture and dual output capabilities. Coupled with ligase chain reaction (LCR), this system facilitates highly sensitive and specific detection of single nucleotide polymorphisms (SNPs). Specifically, LCR precisely identifies single-base mutations, effectively transmitting biological information. The 3D DNA walker system is based on entropy-driven circuit cycling reaction technology. In this system, LCR products serve as the driving strands for the DNA walker, independently binding to track strands and walking legs immobilized on gold nanoparticles, forming a unique dual signal capture mechanism. Each track strand carries two signal chains, significantly enhancing signal amplification efficiency. Benefiting from this novel enzyme-free 3D DNA walker strategy, our biosensing system exhibits exceptional sensitivity to mutant targets (MT), detecting MT at concentrations as low as 30.3 aM and distinguishing heterozygous samples with a 0.01% mutation frequency. Furthermore, this system has been successfully applied to genotyping and mutation abundance assessment of genomes from fresh soybean leaves, demonstrating its vast potential for practical applications. In summary, this research pioneers a novel enzyme-free 3D DNA walker with dual capture and dual output capabilities, and develops an ultrasensitive genotyping tool. This provides strong technical support for the advancement of genetic research.
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JOURNAL OF THE ELECTROCHEMICAL SOCIETY
·
2025
·Vol. 172
·期 4
·045502
·
IF 3.3
Lingling Zhang,Shuangru Yang,Yuge Liu,Yongmei Jia,Zhiguo Li
Understanding the electron transfer performance of DNA self-assembled monolayers (SAMs) on a transducer substrate was important for constructing DNA electrochemical sensors with high sensitivity. Here we immersed the gold electrodes in mixed solutions comprising thiol-modified stem-loop DNA (SL-DNA) or linear DNA (L-DNA) terminally bound with methylene blue (MB), and aliphatic thiols (HOC6SH, C6SH, HOOCC5SH, H2NC6SH, HOC11SH, C12SH, or HOOCC10SH) to obtain DNA/thiols mixed self-assembled monolayers (SAMs). MB electron transfer performances through SL-DNA/thiols and L-DNA/thiols mixed SAMs were investigated by cyclic voltammetry (CV). Our results showed that the mixed assembly of thiols with DNA was conducive to MB electron transfer. The electron transfer rate (ks) values of MB through L-DNA SAMs were bigger than those through SL-DNA SAMs, indicating that the flexibility of DNA chains was important to MB electron transfer. Thiols chain length or terminal groups influenced ks of MB significantly, that increasing NaCl or MgCl2 concentrations in solution around DNA availed MB electron transfer, and Mg2+ advanced MB electron transfer more than Na+, and that the stability of L-DNA/thiols SAMs was much better than SL-DNA/thiols SAMs. These conclusions might provide a reference for studying DNA electron transfer mechanisms or designing DNA or RNA electrochemical sensors with optimal performance.
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Materials Today Bio
·
2025
·
IF 10.2
Jialu Zhang,Xiaoxuan Wang,Lu Guo,Shan Xiao,Dong Meng,Mengmeng Shang,Xiao Sun,Dandan Shi,Yading Zhao,Rui Liu,Shuting Huang,Xinyu Zeng,Jie Li
PARP inhibitor (PARPi)-based synthetic lethal therapies have displayed limited benefits in BRCA-proficient ovarian cancer. To potentiate the application of PARPi, an ultrasound contrast agent OLA-NDs for delivery of the PARPi olaparib (OLA) was established for enhancing DNA damage by blocking DNA repair. OLA-NDs were endowed with endogenous pH- and exogenous ultrasound (US)-responsiveness to target tumors, as well as contrast-enhanced US imaging for diagnostic and therapeutic integration. OLA-NDs could upregulate NOX4 to induce oxidative stress and sensitize BRCA wild-type A2780 cells to DNA oxidative damage through the utilization of ultrasound-targeted microbubble destruction (UTMD). In addition, the strategy further increased ROS production by interfering with mitochondrial function, thereby exacerbating DNA double-strand breaks (DSBs) and inducing mitochondria-mediated apoptosis. As a consequence, the combined application of UTMD and OLA-NDs demonstrated significant antitumor effects in vitro and in vivo . This combined strategy of amplifying oxidative damage improved lethality by promoting DNA DSBs and apoptosis with reduced adverse side effects, which would provide new insight for the clinical application of PARPi in BRCA-proficient ovarian cancer.
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CURRENT EYE RESEARCH
·
2025
·
IF 2
Xin Zheng,Guo-Jian Jiang,Ting-Jun Fan
Purpose The blue light from the digital screens endangers the visual system among which the corneas at the outmost of eyes are vulnerable to the irradiation. Therein, the human corneal endothelial (HCE) cells are crucial to maintain corneal transparency and their damage leads to HCE decompensation resulting in blindness ultimately. Thus, understanding the phototoxic effects of the blue light on the HCE cells and the underlying mechanisms is important for taking measures to protect the vision clarity from the blue-light hazard.Methods We pulse-irradiated the HCE cell line cells at logarithmic phase for 3 passages using 440 nm blue light and examined the levels of reactive oxygen species (ROS), ATP, nicotinamide adenine dinucleotide (NAD+) and autophagy using cytochemistry assay to investigate the alterations of energy metabolism. Moreover, we examined the γH2AX+ cells using immunofluorescence and expression of poly(ADP-Ribose)polymerase1 (PARP1) using western blotting to investigate the degrees of DNA damage and repair. We also monitored the levels of inflammasome using western blotting and senescence associated secretory phenotypes (SASPs) of interleukin (IL)-8, IL-1β and IL-6 using qPCR and ELISA to investigate the inflammasome assembly and secretion of SASPs. We detected the senescent features with senescence-associated-β-galactosidase assay, p16 levels by western blotting, Lamin B1 localization by immunofluorescence observation, cell growth by EdU incorporation assay and confluence forming time and alterations of the cell morphology and relative areas by microscopy observation.Results The HCE cells exhibited senescent features after blue-light-pulse-irradiation. The blue light provokes overproduction of ROS to decrease the levels of ATP, NAD+ and autophagy leading to energy crisis. Moreover, the excess ROS injure DNA and downregulate PARP1 resulting in stable cell-cycle arrest. The excess ROS also facilitate inflammasome assembly leading to hypersecretion of SASPs.Conclusion The blue light elicits HCE cell senescence via inducing energy crisis, stable cell-cycle arrest and SASP hypersecretion.
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INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2025
·
IF 8.5
Haifeng Wang,Yi Huang,Chenhui Zhou,Fanyong Gong,Jianyong Wang,Hao Liu,Wenhui Zhao,Yuxiang Zhao,Xiang Gao
In recent years, the use of nanocarriers to deliver bioactive peptides has become a new strategy to promote angiogenesis. The aim of this study was to develop a QKCMP peptide delivery system based on triangular DNA origami nanostructure (tDONs) carrier to improve the bioactivity and targeting of the peptide. The main objective of this study was to design and prepare QKCMP peptide-TDOns complexes and to evaluate their angiogenesis promoting effects in vitro and in vivo. tDONs were synthesized and purified, and then QKCMP peptide was loaded into tDONs. A variety of characterization techniques were used to evaluate the physicochemical properties of the complexes, including particle size, morphology, and RNA release efficiency. The effects of QKCMP peptid-TDons on proliferation, apoptosis, migration and angiogenesis of human umbilical vein endothelial cells (HUVEC) were evaluated by CCK8, EdU, Annexin V-FITC/PI, PI staining and Transwell assay. At the same time, its promoting effect on angiogenesis was further verified in zebrafish and mouse models. The successful preparation of QKCMP peptide-TDONS showed good biocompatibility and stability. The results of in vitro experiments showed that QKCMP peptide-TDons significantly promoted HUVEC proliferation, inhibited apoptosis, and activated endothelial cell function. Zebrafish experiments showed that QKCMP peptid-TDons effectively enhanced angiogenesis and promoted rapid repair of carotid vascular endothelium in mouse models.
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ANALYST
·
2025
·
IF 3.3
Jiahui Lai,Danni Wang,Xiuqin Sheng,Chuan-Fan Ding,Yinghua Yan
In large-scale glycoproteomics studies, reducing the experimental duration and enhancing digestion efficiency are crucial. In this work, a magnetic nanomaterial featuring a surface rich in hydrophilic groups and uniformly distributed trypsin (Fe3O4@DOPA/PEI@Au@DNA TET-trypsin) was developed for efficient digestion of proteins and specific enrichment of glycopeptides. The material surface exhibits a regular distribution of trypsin coupled with abundant hydrophilic groups. Thus, the material can demonstrate excellent performance in experimental studies related to glycoproteomics. Fe3O4@DOPA/PEI@Au@DNA TET-trypsin showed an excellent limit of detection (0.001 amol·μL−1) and high selectivity for glycopeptides (BSA : HRP = 5000 : 1) in a short reaction time, and it showed good performance in the processing of practical samples. Gene Ontology (GO) analysis suggests that complement activation, extracellular space, antioxidant activity and complement binding may be associated with tuberculosis. These analytical results provide new insights for determining and clarifying the pathogenesis and pathology related to tuberculosis.
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GIANT
·
2025
·
IF 4.9
Yanrong Shuai,Xiaojuan Miao,Yan Wang,Chaozheng Zhang,Yu Liu,Yuening Cao,Tingting Zhang,Jun Lu,Yilan Liu
Conventional DNA double-strand breaks (DSBs)-inducing chemotherapeutics for acute myeloid leukemia (AML) are often limited by poor solubility, non-selectivity, and drug resistance stemming from robust tumor DNA repair mechanisms. Here, we report a glutathione (GSH)-responsive, dual-targeting nano-prodrug, designated HA-FA@Etp-Olp, for efficient AML cell eradication. The HA-FA@Etp-Olp system was constructed through the conjugation of the PARP inhibitor Olaparib (Olp) with Etoposide (Etp) via a disulfide linkage, forming an Etp-Olp heterodimeric prodrug. This hydrophobic conjugate was encapsulated within a polymeric carrier composed of poly (ethylene glycol)-modified hyaluronic acid (HA) functionalized with folic acid (FA), self-assembling into well-refined nanoparticles. Exhibiting excellent circulatory stability, HA-FA@Etp-Olp achieved efficient accumulation within AML cells leveraging CD44/FR dual-receptor-mediated active targeting, followed by GSH-triggered disassembly and specific drug release in response to elevated intracellular GSH levels. Furthermore, HA-FA@Etp-Olp elicited a synergistic cytotoxic effect against AML through a dual-pronged mechanism: "DNA damage-repair blockade" cascade and significant augmentation of mitochondrial oxidative stress, effectively inducing apoptotic cell death. This strategy provides a promising targeted nanotherapeutic approach with enhanced efficacy and reduced systemic toxicity, demonstrating significant potential for the precise treatment of AML.
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BIOMATERIALS
·
2017
·
IF 8.402
Si-An Xie,Tao Zhang,Jin Wang,Feng Zhao,Yun-Peng Zhang,Wei-Juan Yao,Sung Sik Hur,Yi-Ting Yeh,Wei Pang,Li-Sha Zheng,Yu-Bo Fan,Wei Kong,Xian Wang,Jeng-Jiann Chiu,Jing Zhou
Cells perceive the physical cues such as perturbations of extracellular matrix (ECM) stiffness, and translate these stimuli into biochemical signals controlling various aspects of cell behavior, which contribute to the physiological and pathological processes of multiple organs. In this study, we tested the hypothesis that during arterial stiffening , vascular smooth muscle cells (SMCs) sense the increase of ECM stiffness, which modulates the cellular phenotype through the regulation in DNA methyltransferases 1 (DNMT1) expression. Moreover, we hypothesized that the mechanisms involve intrinsic stiffening and deficiency in contractility of vascular SMCs. Substrate stiffening was mimicked in vitro with polyacrylamide gels. A contractile-to-synthetic phenotypic transition was induced by substrate stiffening in vascular SMCs through the down-regulation of DNMT1 expression. DNMT1 repression was also observed in the tunica media of mice aortas in an acute aortic injury model and a chronic kidney failure model, as well as in the tunica intima of human carotid arteries with calcified atherosclerotic lesions. DNMT1 inhibition facilitates arterial stiffening in vivo and promotes osteogenic transdifferentiation , calcification and cellular stiffening of vascular SMCs in vitro . These effects may be attributable, at least in part, to the role of DNMT1 in regulating the promoter activities of Transgelin (SM22α) and α-smooth muscle actin (SMA) and the functional contractility of SMCs. We conclude that DNMT1 is a critical regulator that negatively regulates arterial stiffening via maintaining the contractile phenotype of vascular SMCs. This research may facilitate elucidation of the complex crosstalk between vascular SMCs and their surrounding matrix in healthy and in pathological conditions and provide new insights into the implications for potential targeting of the phenotypic regulatory mechanisms in material-related therapeutic applications.
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Synthetic and Systems Biotechnology
·
2025
·
IF 4.4
Wen Tian,Songcheng Yu,Kaiyang Zhang,Tao Liu,Lihua Ding,Peng Zhang
The CRISPR/Cas12a system holds significant promise for biomedical applications. Nevertheless, the commonly used reporter, fluorophore–quencher-labeled substrates, is hindered by labor-intensive synthesis procedures and high costs, while also relying on a single-photon method and being vulnerable to environmental interference. Herein, a label-free dual-fluorescent functional nucleic acid (DFFNA) was engineered, comprising an aptamer domain for auramine O (AO) recognition and a dSpacer-integrated DNA duplex region for 5,6,7-trimethyl-1,8-naphthyridin-2-amine (ATMND) binding. The fluorescence of AO and ATMND can be enhanced and quenched, respectively, when bound to DFFNAs. The fluorescence intensity ratio between ATMND and AO increased significantly following the cleavage of DFFNAs by activated Cas12a, thus offering a universal, label-free, ratiometric fluorescent reporter for the CRISPR/Cas12a system. To explore the application of the DFFNA-based CRISPR/Cas12a system, a novel biosensor was developed to detect site-specific DNA methylation. It employs a methylation-sensitive restriction enzyme to recognize methylation sites, Cas12a for site-specific DNA identification and signal amplification, and DFFNAs to produce ratiometric fluorescence. The assay demonstrated remarkable specificity and sensitivity, with a limit of detection of 152 pM, due to the high resolution and trans -cleavage activity of Cas12a. The rationally designed and label-free DFFNAs enhance stability, increase flexibility, and reduce cost. The observable color change and smartphone imaging capability facilitate portable, point-of-care testing. Specifically, the biosensor demonstrated excellent specificity by differentiating colorectal cancer patients from healthy individuals. Consequently, this work presents a superior label-free and ratiometric fluorescent reporter for the CRISPR/Cas12a system, which offers a promising strategy for DNA methylation detection in clinical settings.
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LWT-FOOD SCIENCE AND TECHNOLOGY
·
2025
·
IF 6.6
Jianmei Li,Yuting Yin,Haiyu Li,Zongyao Zhao,Mengqin Xiang,Yong Tang,Qi Sun
The escalating economic and ethical concerns associated with camel meat adulteration using cheaper horsemeat necessitate the development of rapid, on-site detection technologies that overcome the limitations of conventional laboratory-based methods. In this study, a portable electrochemical DNA biosensor was fabricated by integrating a polydopamine (PDA)-modified screen-printed carbon electrode (SPCE) with a nucleic acid-functionalized UIO-66(Zr) metal-organic framework (MOF). The PDA coating facilitated stable, covalent immobilization of DNA capture probes through Schiff base reactions, while the UIO-66(Zr) MOF functioned as a high-capacity signal amplifier owing to its large surface area and intrinsic redox activity. This synergistic architecture enabled ultrasensitive detection, achieving a limit of 3.57 pmol/L for horsemeat DNA and 0.5 g/kg adulteration in camel meat blends—significantly surpassing the performance of existing detection methods. The biosensor exhibited excellent specificity by distinguishing single-base mismatches, along with high reproducibility (RSD = 4.12%), operational stability over 15 days, and accurate recovery rates ranging from 91.7% to 112.4% in spiked samples. Its disposable electrode format, cost-effectiveness, and compatibility with portable potentiostats underscore its strong potential for on-site meat authentication. Overall, this work provides a practical and scalable solution to combat meat fraud in resource-limited settings and establishes a versatile MOF-electrode nanoassembly platform for advanced food safety monitoring.
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危险品化学品经营许可证（带存储）