PESTICIDE BIOCHEMISTRY AND PHYSIOLOGY
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2019
·
IF 2.87
Benshui Shu,Jingjing Zhang,Zhiyan Jiang,Gaofeng Cui,Sethuraman Veeran,Guohua Zhong
Harmine , a useful botanical compound, has demonstrated insecticidal activity against some pests. However, harmine's mechanism of action has not been thoroughly elucidated to date. To preliminarily explore harmine's insecticidal mechanisms, the cytotoxicity of harmine against Spodoptera frugiperda Sf9 cells was comprehensively investigated. Our results indicated that harmine induced apoptosis in Sf9 cells, as evidenced by cellular and nuclear morphological changes, DNA laddering and increases in caspase-3-like activities. In addition, activation of the mitochondrial apoptotic pathway by harmine was confirmed by the generation of ROS, opening of mitochondrial permeability transition pores (MPTPs), increase in cytosolic Ca 2+ , changes in mRNA expression levels of genes involved in the mitochondrial apoptotic pathway and increase and release of Cytochrome c. Furthermore, lysosomal membrane permeabilization, release of cathepsin L from the lysosome into the cytosol and cleavage of caspase-3 were also triggered, which indicated that lysosomes were involved in this physiological process . Moreover, the effect of harmine on DNA topoisomerase I activity was investigated by in vivo and molecular docking experiments. These data not only verified that harmine induced apoptosis via comprehensive activation of the mitochondrial and lysosomal pathways and inhibition of DNA topoisomerase I activity in Sf9 cells but also revealed a mechanism of harmine insecticidal functions for pest control.
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JOURNAL OF PHOTOCHEMISTRY AND PHOTOBIOLOGY A-CHEMISTRY
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2014
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IF 2.291
Li-Na Wen,Meng-Xia Xie
s G-quadruplex DNA is an effective target for anticancer drug, and investigation of the G-quadruplex DNA with natural compounds plays a key role in the development of anticancer drugs. In this article, room temperature phosphorescence sensing system based on l -cysteine-capped Mn-doped ZnS QDs has been established. Sanguinarine can adsorb on the surface of Mn-doped QDs and quenched the phosphorescence emission of the sensor. When the G-quadruplex sequences combined with the sanguinarine of the QDs-sanguinarine system, the phosphorescence intensities of the QDs sensors would be restored, and the binding abilities of the G-quadruplex DNA with the sanguinarine can be evaluated by the phosphorescence recoveries of the sensor system. Based on the developed approach, the interaction of sanguinarine and six G-quadruplex DNA with various sequences (tel24-1, tel24-2, tel22, k-ras, HIF-1α and c-kit1) has been investigated. The results showed that the binding abilities of the G-quadruplex DNA with sanguinarine were related to their number of bases and conformations. The conformational changes of the six G-quadruplex DNA, after binding with sanguinarine, have also been probed by circular dichroism. Combining the changes of the phosphorescence lifetime, the binding mechanism of the G-quadruplex DNA and sanguinarine has been discussed. The developed method was simple, rapid and specific, and can avoid fluorescence interferences of the system, and it would be prospective to evaluate the interaction of the drug with different G-quadruplexes.
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ANALYST
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2016
·Vol. 141
·期 5
·1830-1836
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IF 4.033
Xiuzhong Wang,Shanshan Dong,Ting Hou,Lei Liu,Xiaojuan Liu,Feng Li
A novel enzyme inhibition-based homogeneous electrochemical biosensing strategy was designed for an organophosphorus pesticide assay based on exploiting the resistance of a mercury ion-mediated helper probe (HP) toward nuclease-catalyzed digestion and the remarkable diffusivity difference between HPs and the mononucleotides toward a negatively charged indium tin oxide (ITO) electrode. In particular, the mercury ion-mediated T–Hg2+–T base pairs facilitate the HP labeled with methylene blue (MB) to fold into a hairpin structure, preventing its digestion by exonuclease I, and thus resulting in a low electrochemical response because of the large electrostatic repulsion between the negatively charged ITO electrode and the HPs. The competitive binding by a thiol group (–SH), produced in the hydrolysis reaction of acetylthiocholine (ACh) chloride with acetylcholinesterase (AChE), removes mercury ions from the base pairs, causing a nuclease-catalyzed digestion, and the subsequent electrochemical response increase due to the weak electrostatic repulsion between the product-mononucleotides and the ITO electrode. Mercury ion-mediated HPs were first designed for pesticide detection and diazinon was chosen as the model target. Under the optimal experimental conditions, the approach exhibited high sensitivity for diazinon detection with a detection limit of 0.25 μg L−1. The satisfactory results in the determination of diazinon in real samples demonstrate that the method possesses great potential for detecting organophosphorus pesticides. This new approach is expected to promote the exploitation of mercury-mediated base pair-based homogenous electrochemical biosensors in biochemical studies and in the food safety field.
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MICROCHIMICA ACTA
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2019
·Vol. 186
·期 9
·1-7
·
IF 5.479
Zhang Pu,Jia Chunyan,Zhao Yannan,Luo Honghong,Tan Xin,Ma Xiaohong,Wang Yi
Tiopronin is a widely used drug for treatment of cystinuria, rheumatoid arthritis and hepatic disorders. It is also an antidote to heavy metal poisoning and a radioprotective agent. A method is described for rapid and sensitive determination of tiopronin using DNA-stabilized silver nanoclusters (DNA–AgNCs) as a fluorescent probe. Tiopronin can selectively bind to DNA–AgNCs to form a stable Ag-S bond upon which the red photoluminescence (best measured at excitation/emission wavelengths of 590/640 nm) is quenched. The finding is used to design an assay that has a linear response in the 1–150 nM tiopronin concentration range and a 270 pM limit of detection. Compared with previously reported methods, the present approach is more rapid, highly sensitive and selective. It has been successfully applied in the detection of tiopronin in spiked urine and serum, and in pharmaceutical products (tablets and injections).
An ultrasensitive and reliable method for tiopronin assay is developed using red-emissive silver nanoclusters as a fluorescent probe. It has been successfully applied in the determination of tiopronin in biological fluids and pharmaceutical products.
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JOURNAL OF FLUORESCENCE
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2013
·Vol. 24
·期 2
·533-539
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IF 1.789
Liu Chengxuan,Liang Gaiting,Liu Zhen,Zu Lily
Time-resolved fluorescence anisotropy spectroscopy was applied to study the interaction between a peptide truncated from the binding site of tumor suppressor p53 protein and the DNAs covalently labeled with 6-carboxyfluorescein (FAM) dye. Fluorescence intensity quenching and changes of anisotropy decay lifetime were monitored when FAM labeled DNA formed complex with the peptide. The results demonstrated that the sequence of DNA could not define the binding specificity between the peptide and DNA. But the anisotropy decay of FAM can be used to examine the binding affinity of the peptide to DNA. The fluorescent dynamics of FAM can also be used to represent the rigidity of the complex formed between the peptide and DNA.
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LWT-FOOD SCIENCE AND TECHNOLOGY
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2020
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IF 4.006
Ting Liu,Jiamu Kang,Liu Liu
Pseudomonas aeruginosa is a notoriously difficult pathogen to control with antibiotics and disinfectants. Thymus vulgaris L. essential oil (TEO) and thymol are useful for replacing synthetic bacteriostatic agents due to their broad-spectrum bactericidal properties. This study focuses on understanding the inhibitory effects of TEO and its major component against P. aeruginosa , and exploring the antibacterial mechanism from the perspectives of cell membrane destruction, DNA intercalation and biofilm prevention. Results indicate that thymol is the principal component in TEO, as confirmed by GC-MS analysis. TEO and thymol potently combat the planktonic P. aeruginosa and directly change the cell structure. P. aeruginosa cell membrane integrity is destroyed as evidenced by an increase in permeability of the inner and outer membranes and the leakage of intracellular biological macromolecules (DNA). Our study further confirms that the accumulation of intracellular ROS caused by TEO and thymol can result in oxidative stress-mediated DNA damage. Thymol intercalates in the P. aeruginosa genomic DNA to affect the normal function of DNA and even cause bacterial death. Moreover, TEO and thymol affect the biofilm formation and limit the bacterial swimming and swarming motilities. This study provides a potential approach for the development and utilization of natural essential oils and bioactive compunds as novel food additives in the food processing.
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Nanoscale
·
2016
·Vol. 8
·期 34
·15681-15688
·
IF 7.76
Siqi Zhang,Kun Wang,Congcong Huang,Zhenyu Li,Ting Sun,De-Man Han
A series of multiple logic circuits based on magnetic beads and DNA are constructed to perform resettable nonarithmetic functions, including a digital comparator, 4-to-2 encoder and 2-to-3 decoder, 2-to-1 encoder and 1-to-2 decoder. The signal reporter is composed of a G-quadruplex/NMM complex and a AuNP-surface immobilized molecular beacon. It is the first time that the designed DNA-based nonarithmetic nanodevices can share the same DNA platform with a reset function, which has great potential application in information processing at the molecular level. Another novel feature of the designed system is that the developed nanodevices are operated on a simple DNA/magnetic bead platform and share a constant threshold setpoint without the assistance of any negative logic conversion. The reset function is realized by heating the output system and the magnetic separation of the computing modules. Due to the biocompatibility and design flexibility of DNA, these investigations may provide a new route towards the development of resettable advanced logic circuits in biological and biomedical fields.
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RSC Advances
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2017
·Vol. 7
·期 22
·13177-13183
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IF 3.108
Yunyun Huang,Bo Yu,Tuan Guo,Bai-Ou Guan
Ultrasensitive and in situ DNA detection at different pH values, ranging from 4.3 to 8.5, based on a microfiber with a graphene oxide linking layer was proposed and experimentally demonstrated. The graphene oxide coating over the silica microfiber provides a strong π–π interaction with the detected single-stranded DNA molecules, and induces an amplified surface refractive index modulation over the fiber, and finally leads to a wavelength shift of the optical interference fringe for interrogation. Using this strategy, in situ DNA measurement with a detection limit of up to 10−12 M and a linear response in the pH range from 4.3–8.5 has been achieved. Benefiting from its compact size, high sensitivity, and ease of use, together with remote operational ability, the proposed sensor opens up a multitude of opportunities for quantifying DNA in various hard-to-reach environments. It may supplement the existing DNA detection tools.
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BIOMACROMOLECULES
·
2025
·
IF 5.4
Lin Zhang,Jiaxing Zhang,Lvping Wu,Shengping You,Rongxin Su,Wei Qi
Single-stranded DNA-binding protein gp32 is essential for DNA replication and widely used in isothermal amplification techniques such as recombinase polymerase amplification (RPA). However, its inherent thermal instability and reaction condition sensitivity restrict its broader applications. Here we develop the Fermentation Integrated Rational Engineering (FIRE-3S) strategy to enhance the functional properties of gp32 through molecular dynamics (MD) simulations, structure-guided fermentation tuning and stability-focused rational design. Starting from a disulfide-bonded mutant (V62C/T80C) that showed enhanced RPA performance but reduced thermostability, MD simulations revealed local structural disruption near the Zn2+-binding motif. Then, structure-guided fermentation tuning was applied, which restored structural integrity and enhanced DNA-binding affinity. Building on 2S-step, a computationally guided clique-based design pipeline rapidly identified a multisite mutant, gp32-M1, with elevated thermostability (ΔTm = +6.03 °C), robust ssDNA-binding, and strong inhibitor tolerance─maintaining >65% activity under 4% isopropanol or ethanol. This work establishes a systematic framework for engineering ssDNA-binding proteins with enhanced biostability and environmental resilience for molecular diagnostics.
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ANALYTICAL CHEMISTRY
·
2025
·
IF 6.7
Chen Huang,Ling-Jie Guo,Jia-Yi Li,Yue-Wen Chen,Bing Wu,Yi-Wei Liu,Chen-Zhi Zhang,Wei-Wei Lin,Ye Yang,Jin-Yuan Chen,Zhou-Jie Liu
The combination of CRISPR-Cas12a with aptamers can potentially improve the sensitivity, specificity, and speed of non-nucleic acid target detection. Nevertheless, current CRISPR-Cas12a aptasensors, solely dependent on aptamer affinity and overlooking the enzymatic regulation of CRISPR-Cas12a, may produce false-positive signals. We proposed a CRISPR-Cas12a aptasensor synergistically regulated by an aptamer and RNA, which incorporated aptamer-mediated drug recognition and CRISPR-powered signal amplification in a one-pot format. Herein, a regulatory RNA probe synergized with the conformational switching of a drug-induced aptamer, enabling precise regulation of Cas12a trans-cleavage activity via toehold-mediated strand displacement (TMSD). This dual regulatory mechanism transformed the aptamer-activated CRISPR-Cas12a sensing process into a TMSD-driven conditional reaction, avoiding false-positive signals and thus achieving better detection performance with a lower detection limit. With the vancomycin (VCM) aptamer as a model, the aptasensor can detect VCM within 30 min from 2% serum, 1% synovial fluid, and 1% cerebrospinal fluid, with a detection limit of 13.62, 7.56, and 6.75 nM, respectively. The proposed aptasensor was incorporated into a custom 3D-printed portable fluorometer and underwent clinical validation in 22 VCM serum samples, reporting no significant difference when compared with the enzyme-multiplied immunoassay technique, confirming the reliability for point-of-care quantification. It further received cross-validation with a quinine aptamer, suggesting universality [a linear range of 10–250 nM (R2 = 0.985) and a detection limit of 0.42 nM]. By integration of aptamer selectivity with CRISPR programmability, this work presents a novel robust biosensing paradigm for point-of-care drug concentration detection.
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ADVANCED FUNCTIONAL MATERIALS
·
2026
·
IF 19.9
Yi Yuan,Qiufeng Song,Linwen Lan,Xuefen Chen,Jiangchuan Du,Linjun Zhang,Nan Chen,Zhifa Shen,Chang Xue
Chemotherapy remains the cornerstone of malignant tumor treatment; however, off-target effects and severe systemic toxicity limit its efficacy. Here, we developed a programmable theranostic platform based on an all-in-one smart DNA nanodevice (SND) that integrates a programmable cancer-cell classifier (PCC) with a responsive delivery system (RDS). By employing a modular design that permits the customization of aptamer combinations for specific cancer cell types, the platform achieves accurate tumor recognition and spatially controlled drug release. Functioning as a dual-aptamer molecular guide, the PCC utilizes an interlocked configuration to identify cell-surface biomarkers in a stepwise manner, thereby ensuring highly specific target cell recognition. The RDS is constructed from a 1D central trunk flanked by double-stranded drug-loading units, a structure that confers high payload capacity and nuclease-enhanced resistance. These peripheral units also function as recognition elements for endogenous stimuli; upon activation, they trigger conformational changes that facilitate efficient intracellular drug release. In vivo studies in tumor-bearing mice demonstrate that this programmable theranostic platform selectively accumulates in tumor tissues, leading to marked inhibition of tumor growth and a substantial reduction in systemic toxicity. By integrating programmable molecular recognition with stimulus-responsive drug delivery, our theranostic platform offers a promising strategy for advancing targeted cancer therapy.
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ANALYTICAL AND BIOANALYTICAL CHEMISTRY
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2026
·
IF 3.8
Yin Yanjun,Shen Hailin,Wang Mengyao,Suo Zhiguang,Liu Yong,Wei Min
A self-calibrated ratiometric electrochemical aptasensor was developed for the detection of ochratoxin A. This sensor is based on DNA-mediated gold/prussian blue nanoflowers (Au/PB NFs), methylene blue/platinum/zinc-cobalt metal-organic framework (MB/Pt/ZnCo MOF), and an exonuclease III (Exo III)–driven bipedal DNA walker cyclic amplification strategy. The self-calibration mechanism of this sensor relies on the ratio between the response signal of MB/Pt/ZnCo MOF (I MB ) and the internal reference signal of Au/PB NFs (I PB ). This ratio can effectively resist external interference factors. DNA-mediated Au/PB NFs with a flower-like structure provide stable internal reference signals and promote electron transfer. MB/Pt/ZnCo MOF, leveraging ZnCo MOF with a porous structure and bimetallic synergistic effects, and Pt with electrocatalytic activity, serve as response signals. An Exo III–driven biped DNA walker was used to realize the cyclic amplification strategy. The sensor showed a wide linear range of 0.001~500 ng/mL, with a low limit of detection of 0.17 pg/mL. The detection capability of spiked corn flour, wine, and coffee powder samples was satisfactory, and the detection results for the quality control samples of coarse rice flour and wheat flour were not significantly different from those of high-performance liquid chromatography (HPLC), showing the potential for practical application.
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