ANALYST
·
2019
·Vol. 144
·期 10
·3389-3397
·
IF 4.019
Jingfeng Wang,Yu Wang,Su Liu,Haiwang Wang,Xue Zhang,Xiaolei Song,Jinghua Yu,Jiadong Huang
DNA can be configured into unique high-order structures due to its significantly high programmability, such as a three-way junction-based structure (denoted Y-shaped DNA), for further applications. Herein, we report a label-free fluorescent signal-on biosensor based on the target-driven primer remodeling rolling circle amplification (RCA)-activated multisite-catalytic hairpin assembly (CHA) enabling the concurrent formation of Y-shaped DNA nanotorches (Y-DNTs) for ultrasensitive detection of ochratoxin A (OTA). Two kinds of masterfully-designed probes, termed Complex I and II, were pre-prepared by the combination of a circular template (CT) with an OTA aptamer (S1), a substrate probe (S2) and hairpin probe 1 (HP1), respectively. Target OTA specifically binds to Complex I, resulting in the release of the remnant element in S2 and successive remodeling into a mature primer for RCA by phi29 DNA polymerase, thus a usable primer-CT complex is produced, which actuates primary RCA. Then, numerous Complex II probes can anneal with the first-generation RCA product (RP) with multiple sites to activate the CHA process. With the participation of endonuclease IV (Endo IV) and phi29, HP1 as a pre-primer containing a tetrahydrofuran abasic site mimic (AP site) in Complex II is converted into a mature primer to initiate additional rounds of RCA. So, countless Y-DNTs are formed concurrently containing a G-quadruplex structure that enables the N-methylmesoporphyrin IX (NMM) to be embedded, generating remarkably strong fluorescence signals. The biosensor was demonstrated to enable rapid and accurate highly efficient and selective detection of OTA with an improved detection limit of as low as 0.0002 ng mL−1 and a widened dynamic range of over 4 orders of magnitude. Meanwhile, this method was proven to be capable of being used to analyze actual samples. Therefore, this proposed strategy may be established as a useful and practical platform for the ultrasensitive detection of mycotoxins in food safety testing.
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FOOD RESEARCH INTERNATIONAL
·
2022
·
IF 7.425
Li Zou,Manjun Zhang,Mengyan Li,Zhiyou Xiao,Liansheng Ling
This work developed an enzyme-free fluorescent aptasensor for sensitive aflatoxin B1 (AFB1) detection based on a dual signal amplification strategy of hybridization chain reaction (HCR) and Zn 2+ -dependent DNAzyme. In the presence of AFB1, the aptamer specifically binds to the target, releasing the blocking DNA, which can initiate HCR between hairpin probes H1 and H2. With the addition of the substrate strand (Zn-Sub) and enzyme strand (Zn-Enz) of DNAzyme, HCR product can hybridize with Zn-Sub and Zn-Enz to form triplex DNA and Y-shaped structure together, which further activates the DNAzyme to cleave Zn-Sub. Then, two separated fragments of Zn-Sub respectively hybridize with the fluorescent probe and quencher probe, which results in a dramatic increase in the fluorescence intensity. The proposed aptasensor shows high sensitivity and selectivity for AFB1 detection with a detection limit of 0.22 nmol/L in a linear range of 0.4–16 nmol/L. Moreover, the fluorescent aptasensor exhibits acceptable applicability for detecting AFB1 in oil samples with satisfactory recoveries of 92.2–107.8%. Results are also in agreement with those of the ELISA method, indicating that the fluorescent sensing strategy has great potential applications in food safety control.
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JOURNAL OF NANOBIOTECHNOLOGY
·
2022
·Vol. 20
·期 1
·1-19
·
IF 9.429
Wang Ning,Yu Chang,Xu Tingting,Yao Dan,Zhu Lingye,Shen Zhifa,Huang Xiaoying
Background As the most common subtype in lung cancer, the precise and efficient treatment for non-small cell lung cancer (NSCLC) remains an outstanding challenge owing to early metastasis and poor prognosis. Chemotherapy, the most commonly used treatment modality, is a difficult choice for many cancer patients due to insufficient drug accumulation in tumor sites and severe systemic side-effects. In this study, we constructed a cell-specific aptamer-modified DNA nanostructure (Apt-NS) as a targeting drug delivery system achieving the precision therapy for lung cancer.Methods The synthesis of DNA nanostructure and its stability were evaluated using gel electrophoresis. The targeting properties and internalization mechanism were investigated via flow cytometry and confocal analyses. Drug loading, release, and targeted drug delivery were determined by fluorescence detection, Zeta potentials assay, and confocal imaging. CCK8 assays, colony formation, cell apoptosis, metastasis analyses and in vivo experiments were conducted to assess the biological functions of DNA nanostructure.Results Self-assembled DNA nanoparticles (Apt-NS) had excellent stability to serum and DNase I and the ability to specifically recognize A549 cells. Upon specific binding, the drug-loaded nanoparticles (Apt-NS-DOX) were internalized into target cells by clathrin-mediated endocytosis. Subsequently, DOX could be released from Apt-NS-DOX based on the degradation of the lysosome. Apt-NS-DOX exerted significant suppression of cell proliferation, invasion and migration, and also enhanced cell apoptosis due to the excellent performance of drug delivery and intracellular release, while maintaining a superior biosafety. In addition, the antitumor effects of Apt-NS-DOX were further confirmed using in vivo models.Conclusion sOur study provided cell-specific aptamer-modified DNA nanostructures as a drug-delivery system targeting A549 cells, which could precisely and efficiently transport chemotherapeutic drug into tumor cells, exerting enhanced antineoplastic efficacy. These findings highlight that DNA nanostructure serving as an ideal drug delivery system in cancer treatment appears great promise in biomedical applications.
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MICROCHIMICA ACTA
·
2017
·Vol. 184
·期 5
·1305-1313
·
IF 4.58
Zhou Lili,Wang Jian,Chen Zhongping,Li Junlong,Wang Ting,Zhang Zhang,Xie Guoming
MicroRNAs (miRNAs) play a considerable role in cancer occurrence and development, and have been identified as promising noninvasive biomarkers. The authors describe a voltammetric method for the determination of the cancer biomarker microRNA-21 (miRNA). It is based on a combination of a universal DNA signal transducer and isothermal target recycling amplification. A hairpin capture probe is bound to the target miRNA to form a duplex structure and to create a toehold in the transducer for initiating the target recycling amplification reaction. In contrast to traditional capture probes, a mismatched site is introduced to improve its ability to capture the target. In order to reduce the complex design procedures of the sequence and widen the applicability of this method, a signal transducer is introduced. Under optimal conditions, response to target miRNA is linear in the 0.5 to 2000 pM concentration range, with a 56 fM. detection limit (at an S/N ratio of 3). In order to characterize the process of target recycling and the stepwise modification of the electrode, real-time fluorescence, agarose gel electrophoresis, cyclic voltammetry, electrochemical impedance spectroscopy and chronocoulometry were used. The results indicate that this isothermal target recycling amplification results in an electrochemical biosensing scheme with wide potential for sensing other bioanalytes.
Schematic illustration of the electrochemical biosensing platform for miRNA-21 detection based on isothermal target recycling amplification and DNA signal transducer triggered strategy.
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MICROCHIMICA ACTA
·
2019
·Vol. 186
·期 8
·1-10
·
IF 5.479
Li Xiaohua,Wang Shanshan,Meng Yuchan,Wang Xiao,Zhang Yue,Hun Xu
Copper(II) polyphthalocyanine (CuPPc) was combined with graphitic carbon nitride (g-C 3 N 4 ) to form a heterojunction with enhanced photoelectrochemical (PEC) signal. A sensitive PEC method was developed for determination of ractopamine based on a PEC inner filter effect between gold nanoparticles (AuNPs) and the g-C 3 N 4 /CuPPc. A gold electrode was modified with g-C 3 N 4 /CuPPc and the DNA was linked to the AuNPs. Initially, the PEC signal is weak due to the inner filter effect between the AuNPs and g-C 3 N 4 /CuPPc. In the presence of ractopamine, it interacts with the aptamer and the complementary chain (C chain) is released. This triggers the entropy-driven cyclic amplification and results in the release of the substrate B chain (SB chain) from three-dimensional DNA stabilizer. The probe is released from the electrode due to the interaction of probe DNA and the SB chain. As a result, the PEC signal increases linearly in the 0.1 pmol·L −1 to 1000 pmol·L −1 ractopamine concentration range. The detection limit is 0.03 pM, and the relative standard deviation is 3.4% (at a 10 pmol·L −1 level; for n = 11). The method has been successfully applied to the determination of ractopamine in pork samples.
Schematic presentation of detection method based on PEC inner filter effect between AuNPs and the g-C 3 N 4 /CuPPc being fabricated for ractopamine. 3D DNA was used as stabilizer to decrease the PEC blank signal.
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BIOORGANIC CHEMISTRY
·
2023
·
IF 5.1
Rui Li,Weiheng Zhao,Chen Jin,Huihua Xiong
Platinum(II)–based drugs play an important role in many chemotherapeutic protocols, but their further clinical applications are hindered by the development of drug resistance and serious side effects. Therefore, to reverse cisplatin (CDDP) resistance in tandem with reduced side effects, nine novel platinum(IV) complexes modified with key pharmacophore of Olaparib were synthesized and evaluated for biological activities. Among them, the optimal complex 8–2 showed good inhibitory activity against PARP–1 and superior anticancer effects over CDDP on parental (MDA–MB–231, IC 50 = 1.13 μM) and CDDP –resistant triple–negative breast cancer (TNBC) cell line (MDA–MB–231/CDDP, IC 50 = 1.72 μM). Detailed mechanisms revealed that compared with Olaparib and CDDP, the enhanced intracellular accumulation of 8–2 could efficiently reverse CDDP resistance in MDA–MB–231/CDDP cells via inhibiting DNA repair–associated mechanisms, enhancing DNA damage, and activating mitochondrion–dependent apoptosis pathway. Furthermore, 8–2 obtained higher tumor growth inhibition rate (64.1 %) than CDDP (26.5 %) in MDA–MB–231/CDDP xenografts, but it did not induce significant toxicity in vivo and in intro , making it a potential drug candidate for the treatment of TNBC.
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TALANTA
·
2023
·
IF 6.1
Lili Chen,Shuang You,Xiaotong Wang,Dong Li,Shuna Ren,Lihua Chen
This study aimed to prepare two hairpin-structure DNA probes by conjugating carminic acid (CA) or hemin into two ends of specific genes of coxsackievirus A16 (CV-A16) and enterovirus A71 (EV-A71) (probe CV-A16-CA and probe EV-A71-hemin ). Then, probe CV-A16-CA and probe EV-A71-hemin as the signal molecules were adsorbed onto NH 2 -MIL-53 (Al) (MOF). Based on these biocomposites, an electrochemical biosensor with dual-signal outputs for simultaneous assay of CV-A16 and EV-A71 was constructed. The stem-loops of probes switched both CA and hemin monomer to dimer, reducing the electrical activity of both CA and hemin. Subsequently, the target-induced opening of the stem-loop switched both CA and hemin dimers to monomers, resulting in two nonoverlapping increasing electrical signals. This sensitively reflected the concentration of target CV-A16 and target EV-A17 ranging from 10 −10 to 10 −15 M with a detection limit of 0.19 and 0.24 fM. This strategy was mainly applied to the simultaneous determination of target CV-A16 and target EV-A17 in 100% serum with satisfactory results. The MOF combined with the high loading capacity broke through the intrinsic limitation on sensitivity using the traditional methods. An increase of three orders of magnitude was observed. This study involved simple one-step detection, and only a simple replacement of a gene could trigger its potential in clinical and diagnostic applications.
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TALANTA
·
2025
·
IF 6.1
Xiang Li,Yuting Zhang,Mengyuan He,Jiarui Sun,Bowen Xiong,Gongke Wang
In this work, we present an ultrasensitive, specific, and high-signal-to-background ratio fluorescence split-aptasensor for D-vasopressin (D-VP) detection. This sensor is based on target-induced self-propelled 3D DNA walkers in conjunction with CRISPR-Cas12a technology. Two split probes (SDA 1 and SDA 2) were designed to undergo structural recombination and function as a walking chain (SDA) under the induction of D-VP. Simultaneously, an intact Mg 2+ -dependent DNAzyme domain was formed at the tail of SDA and subsequently activated. The activated Mg 2+ -dependent DNAzyme continuously propelled the 3D DNA walker, enabling the generation of signal strand DNA (activator DNA). The activator DNA can subsequently trigger the activation of the Cas12a protein, enabling it to cleave the FAM-ssDNA-BHQ1 substrate. This process leads to signal amplification and the specific detection of D-VP. Under optimal conditions, the designed split-aptasensor exhibits excellent linearity across a concentration range of 5 ng/mL to 1215 ng/mL, with a detection limit (LOD) as low as 0.22 ng/mL. This split-aptasensor were employed to identify D-VP in human serum and urine samples, yielding highly satisfactory results. This unique design acts as a proof of concept and illustrates considerable promise for the detection of a wide range of analytes.
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SENSORS AND ACTUATORS B-CHEMICAL
·
2025
·
IF 7.7
Jiarong Lin,Ye Deng,Yue Lin,Fang Luo,Zuquan Weng,Jian Wang,Cuiying Lin,Bin Qiu,Zhenyu Lin
In this study, we present the innovative design and comprehensive evaluation of a novel point-of-care testing (POCT) methodology for the rapid and accurate detection of T-2 toxin, a potent mycotoxin with significant implications for food safety and human health. The cornerstone of this approach lies in the integration of a copper-based conductive metal-organic framework (Cu 3 (HHTP) 2 ) with a target-responsive DNA hydrogel system, creating a dual-signal readout mechanism that significantly enhances detection sensitivity and specificity. Specifically, the hydrogel covers the material's surface metal active sites, which become exposed upon target addition due to hydrogel collapse. The released Cu 3 (HHTP) 2 can catalyze the oxidation of 3,3′,5,5′-tetramethylbenzidine, resulting in a colorimetric and temperature change in the solution. This dual-mode detection strategy enables both qualitative assessment through direct visual inspection of color change and quantitative analysis by monitoring the solution temperature variation post laser irradiation with a thermometer. Under the optimized conditions, the detection system demonstrates a wide range spanning from 5 to 200 ng/mL, with a detection limit of 1.67 ng/mL. This method has been successfully demonstrated through the analysis of real-world samples, yielding encouraging results that underscore its reliability and effectiveness. The proposed dual-signal approach boasts advantages such as straightforward operational procedures, unambiguous signal outputs, and robust practicality, making it an attractive option for mycotoxin detection in resource-constrained settings and developing regions.
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JOURNAL OF NANOBIOTECHNOLOGY
·
2025
·Vol. 23
·期 1
·1-20
·
IF 12.6
Xie Hanbin,Wei Cong,Xiong Chang,Huang Ziyan,Chen Chaojin,Xiao Xue,Zhang Linan,Lin Zhenjia,Yao Weifeng,Zhao Tianyu,Hei Ziqing
Intestinal ischemia/reperfusion (I/R) injury is a common life-threatening condition. Inflammatory dysregulation plays a crucial role in the pathological progression of intestinal I/R injury, indicating that controlling excessive inflammatory responses can be an effective strategy for mitigating I/R injury. Herein, after establishing a correlation between cell-free DNA (cfDNA) levels and postoperative inflammatory factors in samples from patients with intestinal I/R, we tested a cfDNA-scavenging approach for the treatment of intestinal I/R injury. Poly-ε-caprolactone (PCL) microspheres (Micro DEA2k) functionalized with a pH-responsive cationic polymer (DEA2k) to efficiently scavenge cfDNA were synthesized and evaluated.These microspheres exhibited enhanced cfDNA adsorption under inflammation-induced acidic conditions, along with low toxicity, reduced non-specific protein binding, and extended peritoneal retention. In a mouse model of intestinal I/R injury, the intraperitoneal injection Micro DEA2k effectively bound cfDNA, regulated the mononuclear phagocytic system, decreased the number of M1 macrophages, suppressed inflammation, and significantly improved the survival rate of the mice. These findings suggest that cfDNA scavenging using cationic microspheres has considerable potential for alleviating intestinal I/R injury. Graphical
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MICROCHIMICA ACTA
·
2021
·Vol. 188
·期 10
·1-8
·
IF 5.833
Ma Gui-min,Huo Li-wei,Tong Yin-xia,Wang Yu-cong,Li Cui-ping,Jia
Hong-xia
Abstract A novel strategy for microRNAs (miRNAs) detection has been developed utilizing duplex-specific nuclease-assisted signal amplification (DSNSA) and guanine-rich DNA-enhanced fluorescence of DNA-templated silver nanoclusters (AgNCs). The combination between target miRNA, DSNSA, and AgNCs is achieved by the unique design of DNA sequences. Target miRNA opens the hairpin structure of the Hairpin DNA probe (HP) by hybridizing with the HP and initiates the duplex-specific nuclease-assisted signal amplification (DSNSA) reaction. The DSNSA reaction generates the release of the guanine-rich DNA sequence, which can turn on the fluorescence of the dark AgNCs by hybridizing with the DNA template of the dark AgNCs. The fluorescence intensity of AgNCs corresponds to the dosage of the target miRNA. This is measured at 630 nm by exciting at 560 nm. The constructed method exhibits a low detection limit (~8.3 fmol), a great dynamic range of more than three orders of magnitude, and excellent selectivity. Moreover, it has a good performance for miR-21 detection in complex biological samples. Graphical abstract
A novel strategy for microRNAs (miRNAs) detection has been developed utilizing duplex-specific nuclease-assisted signal amplification (DSNSA) and guanine-rich DNA-enhanced fluorescence of DNA-templated silver nanoclusters (AgNCs).
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RSC Medicinal Chemistry
·
2024
·
IF 4.1
Qi Wang,Si-Min Liang,Zhi-Chen Mao,Xian-Li Ma,Jian-Hua Wei,Ri-Zhen Huang,Ye Zhang
A set of biotin-polyethylene glycol (PEG)-naphthalimide derivatives 4a–4h with dual targeting of ferroptosis and DNA were designed and optimized using docking simulation as antitumor agents. Docking simulation optimization results indicated that biotin-PEG4-piperazine-1,8-naphthalimide 4d should be the best candidate among these designed compounds 4a–4h, and therefore, we synthesized and evaluated it as a novel antitumor agent. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay and MGC-803 and U251 xenograft models identified 4d as a good candidate antitumor agent with potent efficacy and safety profiles, compared with amonafide and temozolomide. The findings of the docking simulations, fluorescence intercalator displacement (FID), western blot, comet, 5-ethynyl-2′-deoxyuridine (EdU), flow cytometry, transmission electron microscopy, and BODIPY-581/591-C11, FerroOrange, and dihydroethidium (DHE) fluorescent probe assays revealed that 4d could induce DNA damage, affect DNA synthesis, and cause cell cycle arrest in the S phase in MGC-803 cells. Also, it could induce lipid peroxidation and thus lead to ferroptosis in MGC-803 cells, indicating that it mainly exerted antitumor effects through dual targeting of ferroptosis and DNA. These results suggested that it was feasible to design, optimize using docking simulation, and evaluate the potency and safety of biotin-PEG-1,8-naphthalimide as a antitumor agent with dual targeting of ferroptosis and DNA, based on a multi-target drug strategy.
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