MICROCHIMICA ACTA
·
2025
·Vol. 192
·期 4
·1-13
·
IF 5.3
Wu Qianyue,Yang Beiying,Wang Yutang,Xiong Yanyan,Zhang Jinhua,Wen Yangping,Rajkovic Andreja
An accurate, sensitive, low-cost, portable, and easy-to-use method for the quantitative detection of Clostridium perfringens (Cp), a zoonotic pathogen widely found in nature and capable of spreading through contaminated food or environments, is essential for epidemiology, prevention, and diagnostics. Here, we have designed a smartphone-based electrochemical DNA biosensor, which utilizes carboxyl-functionalized multi-walled carbon nanotubes (COOH-MWCNTs) as the substrate, L-cysteine (L-Cys) as the linker, and a synergistic modification of the bioelectrode with gold nanoparticles (AuNPs) and polyamidoamine dendrimers (PAMAM). This design enables highly sensitive, low-cost, label-free, and portable detection of the cpb2 gene in Cp. The sensor not only enhances detection performance but also improves convenience and practicality. Differential pulse voltammetry (DPV) was used to monitor the electrochemical signal response to changes in target DNA concentration, enabling the tracking of the DNA hybridization process. Under optimized conditions, the biosensor exhibited a linear detection range from 10⁻ 14 to 10⁻⁸ M, with a detection limit of 1.5 fM. It demonstrated excellent selectivity for the cpb2 gene and was successfully applied to detect variations in cpb2 gene content in wastewater and fecal DNA samples. Compared with the traditional method, the detection time of this method is short, the operation of professional and technical personnel is not required, the instrument is small and portable, and the single detection cost is significantly reduced. This study provides a new strategy for the rapid, portable, and highly sensitive detection of bacterial toxin genes in livestock and aquaculture. Graphical
✓ 已验证
产品关联
DOI
PubMed
高影响因子
LWT-FOOD SCIENCE AND TECHNOLOGY
·
2025
·
IF 6.6
Haibin Liu,Yi Yuan,Qiuyue Wang,Hang Tian,Yingxue Wu,Haijie Li,Hongshuo Chen,Lei Wang,Bingrui Liu
Meat adulteration has become a significant concern due to its potential to diminish the nutritional value and trigger allergic reactions. To address the issue of fraudulent adulteration in beef ( Bos taurus ) products, a visual strategy is proposed for multiplex detection of chicken ( Gallus gallus ), duck ( Anas platyrhynchos ), and pork ( Sus scrofa ) components in beef. First, the DNA could be easily extracted from the meat samples using the microneedle within 1 min and used as recombinase polymerase amplification (RPA) template. Next, three primer pairs specific to chicken, duck, and pork DNA are designed with unique oligonucleotide tags and a spacerC9 to halt polymerase activity, enabling the RPA amplicons to possess a single-stranded DNA tail. These tail sequences precisely hybridize with nanoparticle-labeled detection probes and pre-immobilized capture probes on test lines, generating a colorimetric signal on the lateral flow strip (LFS). The enhanced LFS utilizes three distinct Au/Ag nanoparticles, including Au@Ag (yellow), gold nanoflowers (cyan), and gold nanospheres (magenta), each targeting chicken, duck, and pork DNA, respectively. The test output can be easily interpreted through a “magenta yellow cyan” color code. The sensitivity of the multicolor LFS is further improved by the incorporation of DNA-Au/Ag nanoaggregates. Four functional oligonucleotides with complementary sequences are combined to form DNA-Au/Ag nano aggregates, allowing for increased coupling of Au/Ag and enhancing detection sensitivity. Cross-reactivity tests with ten non-target species demonstrate the excellent specificity of the assay. The method can detect chicken, duck, or pork at levels as low as 0.1 g/kg in simulated adulterated beef samples. The entire process, including rapid DNA extraction (1 min), RPA reaction (15 min), and LFS detection (5 min), is completed within 21 min. This platform is user-friendly, eliminating extensive training and specialized equipment, making it ideal for rapid on-site detection of multiplex adulteration by personnel in various fields.
✓ 已验证
DOI
高影响因子
JOURNAL OF BIOMOLECULAR STRUCTURE & DYNAMICS
·
2019
·
IF 3.31
Yu-Jie Luo,Bao-Li Wang,Song-Bo Kou,Zhen-Yi Lin,Kai-Li Zhou,Yan-Yue Lou,Jie-Hua Shi
The binding characteristics of calf thymus DNA (ct-DNA) with dasatinib (DSTN), a tyrosine kinase inhibitor was assessed through multi-spectroscopic methodologies and viscosity measurement combined with molecular docking as well as DFT calculation to understand the binding mechanism, affinity of DSTN onto ct-DNA, effect of DSTN on ct-DNA conformation, and among others. The results confirmed DSTN bound onto ct-DNA, leading to forming the DSTN–ct-DNA complex with the binding constant of 4.82 × 103 M−1 at 310 K. DSTN preferentially inserted to the minor groove of ct-DNA with rich A-T region, that was the binding mode of DSTN onto ct-DNA was groove binding. The enthalpic change (ΔH0) and entropic change (ΔS0) during the binding process of DSTN with ct-DNA were 128.9 kJ mol−1 and 489.2 J mol−1 K−1, respectively, confirming clearly that the association of DSTN with ct-DNA was an endothermic process and the dominative driven-force was hydrophobic interaction. Meanwhile, the results also indicated that there was a certain extent of electrostatic force and hydrogen bonding, but they maybe play an auxiliary role. The CD measurement results confirmed the alteration in the helical configuration of ct-DNA but almost no change in the base stacking after binding DSTN. The results revealed that there was the obvious change in the conformation, the dipole moment, and the atomic charge distribution of DSTN in the B-DNA complexes, compared with free DSTN, to satisfy the conformational adaptation. From the obtained fronitier molecular orbitals of DSTN, it can be inferred that the nature of DSTN alters with the change of the environment around DSTN.Communicated by Ramaswamy H. Sarma
✓ 已验证
DOI
高影响因子
ANALYTICA CHIMICA ACTA
·
2022
·
IF 6.911
Yang-Yang Hou,Wan-Zhen Xie,Xuecai Tan,Ke-Jing Huang,Jing Xu
A highly sensitivity self-powered biosensor is developed based on T7 exonuclease (T7 Exo) and 3D DNA walker induced rolling circle amplification (RCA) for electrochemical/colorimetric dual-mode detection of microRNA-21 (miRNA-21) with improved reliability. Taking its advantage of fascinating properties, such as high structure defects and good conductivity , graphdiyne is prepared and used to prepare high-performance enzyme biofuel cell. T7 Exo-assisted 3D DNA walker target recognition triggers RCA reaction to obtain a significantly amplified signal response. A capacitor is integrated to the enzyme biofuel cell to further amplify the electrochemical output signal of the self-powered biosensor. In detection system, glucose oxidase catalyzes glucose oxidation to produce hydrogen peroxide, and 3,3′,5,5′-tetramethylbenzidine (TMB) is then catalyzed to generate colored products, so as to achieve the colorimetric detection of the target. Analysis signals of diverse modes are recorded independently. Consequently, detection of microRNA with improved reliability and wider signal response range are achieved by electrochemical/colorimetric dual-mode with detection limits of 0.15 and 33 fM ( S/N = 3) respectively. In addition, the proposed self-powered biosensor successfully applied for the detection of miRNA-21 in human serum samples, confirming its practical applicability in clinical diagnosis. It is powerfully anticipated the proposed self-powered biosensor possesses great potential to be applied to other biomedical domains.
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
·
2020
·Vol. 412
·期 8
·1807-1816
·
IF 3.637
Hu Xin,Liu Man,Liu Yifei,Zhang Jingjing,Tang Yuwen,Pei Hong,Liu Hegang,Chen Shilin,Song Chengwu,Hu Zhigang
Herbal materials have both medicinal and commercial values. As such, accurate species and content identification and verification are necessary to ensure the safe and effective use for medical and commodity purposes. Herein, we introduce a two-step approach for systematic identification and quality evaluation of wild and introduced Anemone flaccida Fr. Schmidt (aka Di Wu) using DNA barcode and ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS/MS). To begin, a precise and rapid identification method based on internal transcribed spacer 2 (ITS2) sequence was developed to ensure the authenticity of ‘Di Wu’ species. Next, the major active components were fully characterized utilizing a targeted profile of oleanane-type triterpenoid saponins, which was established via UPLC-QTOF-MS/MS. As a result, 34 oleanane-type triterpenoid saponins were identified or characterized in ‘Di Wu.’ The qualitative and relative quantitative analysis showed obvious differences between wild and introduced ‘Di Wu.’ Furthermore, dynamic changes in the contents of triterpenoid saponins throughout various harvesting periods were clearly explained and mid-April was identified as the appropriate harvest time. Moreover, results indicate that the contents of five main saponins (anhuienoside E, glycosideSt-I4a, hemsgiganoside B, flaccidoside II, and hederasaponin B) are more appropriate as a quality evaluation indicator than the current quality standard. The two-step approach provides a suitable strategy to evaluate the genuine quality of wild and introduced ‘Di Wu,’ and can be applied to the targeted analysis of other triterpenoid saponin analogues for quality evaluation.
.
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Chemical Biology & Drug Design
·
2020
·Vol. 96
·期 6
·1433-1446
·
IF 2.548
Yibing Guo,Liancheng Xu,Changchun Ling,Tao Yang,Wenjie Zheng,Jin Lv,Qingsong Guo,Bohua Chen
Eighteen new β-carboline-based indole-4,7-quinone derivatives ( 12a–i and 13a–i ) were designed and synthesized, and their in vitro and in vivo antiproliferative activities were studied. Most of target compounds showed strong inhibition on three human tumor cells' proliferation. In particular, the most active compound 13g not only displayed more prominent antiproliferative activities than β-lapachone, a clinical antitumor candidate, but also exerted significant NAD(P)H: quinone-oxidoreductase-1 (NQO1) inhibitory activity and NQO1-dependent cytotoxicity in HT29 cells. Furthermore, 13g dose-dependently induced high ROS levels in HT29 cells, and selectively inhibited cancer cell but not non-tumor colon cell proliferation in vitro. Importantly, 13g promoted HT29 cell apoptosis and DNA damage by regulating relative apoptotic proteins and H2AX expression. Finally, 13g displayed significant growth inhibition of HT29 human colorectal adenocarcinoma xenograft in mice without overt toxicity.
✓ 已验证
DOI
PubMed
高影响因子
MICROCHEMICAL JOURNAL
·
2025
·
IF 5.1
Ying Cheng,Jiaxuan Chen,Jiuhua Song,Jiali Chang,Yang Li,Qiao Zhang,Yifan Feng,Kai Shi,Tao Jiang
Detection of MPXV DNA in urban wastewater is crucial for early warning, rapid response, and effective outbreak control, offering a non-invasive approach to strengthen epidemic preparedness and public health defense. Based on heparin sodium-assisted one-tube recombinase polymerase amplification (RPA)/split gRNA with Cas12a (SCas12a) and phosphorothioate-modified hairpin G-quadruplex (psHG4), we have developed a colorimetric biosensing platform to achieve sensitive and on-site detection of MPXV DNA in urban wastewater (called hoRPA/SCas12a). The MPXV DNA initiates a one-tube RPA/SCas12a reaction facilitated by heparin sodium, subsequently inducing the DNase activity of SCas12a. The loop DNA of psHG4 is cleaved by active SCas12a to release the pre-blocked psG4 sequence, enabling the formation of a psG4/hemin complex with K + and hemin. The complex is capable of initiating the oxidation of 3,3′, 5,5′-tetramethylbenzidine (TMB) to generate the colorimetric substrate TMBox. The portable smartphone was employed to record the RGB value changes of the sample. The hoRPA/SCas12a platform displays a linear range of 10 aM to 100 fM, with a detection limit of 3 aM. Moreover, this platform can selectively and accurately detect MPXV DNA in urban wastewater. The hoRPA/SCas12a platform is vital for early warning, effective public health response, and strengthening wastewater-based epidemiology in emerging infectious disease control.
✓ 已验证
产品关联
DOI
高影响因子
MICROCHIMICA ACTA
·
2018
·Vol. 185
·期 3
·1-9
·
IF 5.705
Zhang Kai,Gan Ning,Hu Futao,Chen Xixue,Li Tianhua,Cao Jinxuan
The authors describe an enzyme-free aptamer-based assay for the determination of the model antibiotic kanamycin (Kana). The method is making use of (a) microfluidic chip electrophoresis; (b) a stirring bar carrying a gold-labeled aptamer probe, and (c) the hybridization chain reaction (HCR) for signal amplification. Firstly, a stirring bar (length: 1 cm; diameter: 0.2 mm) was modified with a large amount of duplex DNA and then hybridized with aptamer and its partially complementary chains (cDNA). In the presence of Kana, the binding between the Kana and aptamer unwinds the duplex structures and releases a corresponding amount of cDNA into the supernatant. The released cDNA triggers the HCR in the presence of H1 and H2 DNA hairpin to produce a large amount of duplex DNA chains with different lengths. At the same time, the amounts of H1 and H2 are reduced. The decreased signal of H1/H2 after several HCR cycles can be used to quantify kana in the 1 pg·mL −1 to 10 ng·mL −1 , with a detection limit of 0.29 pg·mL −1 . The signal is generated by reading the fluorescence, best at excitation/emission maxima of 470/525 nm. The whole detection process takes 3 min only. The assay was employed to the detection of Kana in spiked milk and fish samples. Results are consistent with those of an enzyme linked immunosorbent assay. The assay has high throughput, high selectivity, and high amplification capability.
Schematic of a stirring bar functionalized with gold-labeled aptamer acting as the capture probe. It can capture the target and release primer simultaneously. The primer triggers the hybridization chain reaction inducing the consumption of H1 and H2. After a certain reaction time, the mixture is injected into the MCE platform for microfluidic electrophoretic separation and fluorometric detection.
✓ 已验证
产品关联
DOI
PubMed
高影响因子
TOXICOLOGY
·
2025
·
IF 4.6
Han Gao,Hanyu Gao,Hao Yin
Emodepside, a novel antiparasitic agent, exhibits remarkable efficacy against nematodes. Recent research from a phase 2b clinical trial has demonstrated that the most frequent adverse event in the emodepside treatment group is blurred vision, but the mechanism of blurred vision remains largely unexplored. To provide references for secure medication and prospective therapeutic interventions of emodepside, we conducted an investigation into the cytotoxic effects of emodepside on the corneal stroma and its underlying mechanisms using an in vitro model of human corneal stromal (HCS) cells. The loading of the mini-chromosome maintenance 6 protein (MCM6) onto chromatin at replication origins by chromatin licensing and DNA replication factor 1 (CDT1) constitutes a crucial step in licensing DNA for replication. This study demonstrates that emodepside directly binds to the CDT1-binding domain (CBD) of MCM6, competitively blocking the MCM6-CDT1 interaction as confirmed by modified enzyme-linked immunosorbent assay and drug affinity responsive target stability assays. Molecular docking reveals that emodepside binds to CBD through hydrogen bonds with Lys754, Ile760, and Lys770. In HCS cells, this blockade results in the inhibition of DNA replication licensing and a decrease in DNA synthesis, as evidenced by the ethynyl-deoxyuridine incorporation assay, leading to cell cycle arrest at the G 0 /G 1 phase. Moreover, emodepside-induced cell cycle arrest in HCS cells leads to dose-dependent apoptosis and proliferation inhibition with the IC 50 value being 32.3 μM. In conclusion, this study not only provides new insights into the mechanism of emodepside-induced vision blur but also offers a valuable molecular tool for the research of MCM6-CDT1 interaction.
✓ 已验证
产品关联
DOI
PubMed
高影响因子
PHYTOPATHOLOGY
·
2023
·
IF 3.2
Mr. Jiayu Yin,Ms. Shuang Wu,Ms. Yongli Yang,Dr. Delu Wang,Prof. Yue Ma,Dr. Yongtian Zhao,Prof. Sujitraj Sheth,Miss Honglin Huang,Dr. Baoan Song,Dr. Zhuo Chen
Carvacrol expresses a wide range of biological activities, but the studies of its mechanisms focused on bacteria, mainly involving the destruction of the plasma membrane. In this study, carvacrol exhibited strong antifungal activities against several phytopathogenic fungi and determined a novel antifungal mechanism against Lasiodiplodia theobromae. RNA sequencing indicated that many genes of L. theobromae hyphae were predominately induced by carvacrol, particularly those involved in replication and transcription. Hyperchromic, hypsochromic, and bathochromic effects in the UV-visible absorption spectrum were observed following titration of calf thymus DNA (ctDNA) and carvacrol, which indicated the formation of a DNA–carvacrol complex. Circular dichroism (CD) spectroscopy indicated the response of DNA to carvacrol was similar to 4’, 6-diamidino-2-phenylindole (DAPI), but different from that of ethidium bromide (EB), implying the ionic bonds between carvacrol and ctDNA. Fluorescence spectrum (FS) analysis indicated that carvacrol quenched the fluorescence of double-stranded DNA more than single-stranded DNA, indicating that carvacrol mainly bound to double-stranded DNA. A displacement assay showed that carvacrol reduced the fluorescence intensity of the DNA–DAPI complex through competing with DAPI, but this did not occur for DNA–EB. FS assay revealed that carvacrol bound to AAA sequence on the minor groove of ds-oligonucleotides. The hydroxyl of carvacrol was verified to bind to ctDNA through a comparative test in which structural analogs of carvacrol, including thymol, 4-ethyl-1,2-dimethyl, etc. were analyzed. The current study indicated carvacrol can destruct plasma membranes and bind to the minor groove of DNA, which inhibited the fungal proliferation by disturbing the stability of dsDNA.
✓ 已验证
DOI
高影响因子
MICROCHIMICA ACTA
·
2019
·Vol. 186
·期 6
·1-8
·
IF 5.479
Wang Jing,Li Bingzhi,Lu Qiaoyun,Li Xiaoyun,Weng Chenyuan,Yan Xiaoqiang,Hong Junli,Zhou Xuemin
The authors describe a versatile aptasensing scheme based on the use of polypyrrole nanoparticles (PPyNPs) and DNA-silver nanoclusters (DNA-AgNCs) for multiple target detection. The DNA-AgNCs consist of two functional domains, viz. (a) a nucleation domain for attaching the metal core of the nanoclusters, and (b) a recognition domain which consists of a single-stranded aptamer. In the absence of analytes, the single-strand recognition domain will be absorbed onto the surface of the PPyNPs through π stacking and hydrophobic interactions. As a result, the red fluorescence of the DNA-AgNCs (with excitation/emission peaks at 535/625 nm) is quenched by the PPyNPs. On introducing the analytes, the DNA-AgNCs will bind them. This leads to the desorption of DNA-AgNCs and the recovery of the red fluorescence. Based on the above strategy, a versatile, sensitive and selective aptasensor was established for detection of adenosine, thrombin and interferon-gamma. The method was applied to the detection of the above targets in (spiked) serum samples and gave satisfactory results, with detection limit of 0.58 nM for IFN-γ, 0.39 nM for adenosine, and 2.2 nM for thrombin. The use of PPyNPs results in uniquely low non-specific absorption and in improved analytical results in case of real-sample analysis when compared to previously reported methods.
Schematic illustration of DNA-silver nanoclusters and polypyrrole nanoparticles in an aptasensor for detection of multiple targets.
✓ 已验证
产品关联
DOI
PubMed
高影响因子
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
·
2018
·
IF 3.412
Yongxin Fu,Hui Jin,Xiangning Bu,Rijun Gui
In this work, blue-emitting silver nanoclusters (AgNCs) were prepared in a matrix of single-stranded deoxyribonucleic acid (DNA) on the basis of ambient hydrothermal reactions. DNA acted as the stabilizer or coating agent, and NaBH4 was used as the reducing agent. Through the interactions between rhodamine 6G (Rh6G) and the synthesized DNA-AgNCs, the self-assembled complex of DNA-AgNC–Rh6G was generated. Meanwhile, fluorescence emission of AgNCs was weakened as a result of fluorescence-resonance-energy transfer (FRET) from AgNCs (donor) to Rh6G (acceptor). In the DNA-AgNC–Rh6G complex aqueous suspension, the addition of melamine induced obvious emission recovery of AgNCs and fluorescence decrease of Rh6G, attributable to melamine-induced decomposition of the self-assembled complex and anti-FRET effects. There was a well-plotted linear relationship of ratiometric fluorescence intensities (IAgNCs/IRh6G) versus melamine concentration in the range of 0.1–10 μM, with a low detection limit of 25 nM. Responses of IAgNCs/IRh6G to melamine were highly selective and sensitive over potential interferents. A novel dual-emitting ratiometric fluorescence sensor of melamine was facilely constructed on the basis of the DNA-AgNC–Rh6G complex. In particular, the sensor enabled visual fluorescence detection of melamine both in aqueous solution and on wetted filter paper. Superior detection results of the sensor were experimentally obtained and confirmed its high feasibility for melamine detection in practical samples.
✓ 已验证
DOI
高影响因子
危险品化学品经营许可证（带存储）