LWT-FOOD SCIENCE AND TECHNOLOGY
·
2025
·
IF 6.6
Shanglin Li,Ziyang He,Yonghong Meng,Baoxin Li,Mei Liu
Aflatoxin B1 contamination poses severe threats to food safety, vitally requiring reliable assays. Biosensors are effective but subject to unsatisfactory signal conversion efficiency owing to their usage of the monovalent probes with redundant materials and procedures. Herein, we developed a photothermal aptasensor, which combined multifunctional polyvalent DNA scaffolds with thermal imaging for the portable detection of aflatoxin B1. The aptasensor was conferred with favorable properties of specific recognition, surface modification and signal amplification by the polyvalent DNA scaffolds, thereby showed enhanced analytical performance with simplified procedures. Moreover, the detection potential of the aptasensor was further improved by introducing Cu x S nanoparticles to provide intuitive temperature output for portable devices analysis. Consequently, AFB1 could be quantified in a wide range of 1–500 ng/mL with the detection limit of 0.75 ng/mL, merely using common thermometer. For food samples, the spiked recoveries of the proposed method ranged from 93.5 % to 108.7 % with relative standard deviation from 2.9 % to 6.3 %. These results were in agreement with the high-performance liquid chromatography method, demonstrating the good accuracy of this method. Therefore, this method is expected to be extended as a universal detection strategy for food hazards analysis.
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ANALYTICAL CHEMISTRY
·
2024
·
IF 6.7
Pei Wang,Xiaowei Wei,Luming Shen,Kexin Xu,Zhongting Wen,Nengjiao Gao,Ting Fan,Shenmei Xun,Qingyi Zhu,Xiaojun Qu,Yefei Zhu
Bladder cancer (BC) occurrence and progression are accompanied by alterations in microRNAs (miRNAs) expression levels. Simultaneous detection of multiple miRNAs contributes to the accuracy and reliability of the BC diagnosis. In this work, wrinkled silica nanoparticles (WSNs) were applied as the microreactor for multiplex miRNAs analysis without enzymes or nucleic acid amplification. Conjugated on the surface of WSNs, the S9.6 antibody was adopted as the universal module for binding DNA/miRNA duplexes, regardless of their sequence. Furthermore, single-stranded DNA (ssDNA) was labeled with quantum dots (QDs) for identifying a given miRNA to form QDs-ssDNA/miRNA, which enabled the specific capture of the corresponding QDs on the wrinkled surface of WSNs. Based on the detection of fluorescence signals that were ultimately focused on WSNs, target miRNAs could be sensitively identified to a femtomolar level (5 fM) with a wide dynamic range of up to 6 orders of magnitude. The proposed strategy achieved high specificity to obviously distinguish single-base mutation sequences and possessed multiplex assay capability. Moreover, the assay exhibited excellent practicability in the multiplex detection of miRNAs in clinical serum specimens.
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INTERNATIONAL JOURNAL OF FOOD SCIENCE AND TECHNOLOGY
·
2024
·Vol. 59
·期 9
·6231-6242
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IF 2.6
Mengmeng Zhang,Tianchui Wang,Sixian Ou,Yucong Zou,Xuan Xin
Summary Betaine is a natural antioxidant lacking the ability to scavenge free radicals. Although it has been shown to exert antioxidant function by enhancing the expression of antioxidant enzymes, its mechanism has not yet been elucidated. This study explored the role and mechanism of the transcription factor NF-E2-associated factor 2 (Nrf2)-Kelch-like epichlorohydrin-associated Protein 1 (Keap1) antioxidant reaction element (ARE) pathway in betaine-mediated enhancement of antioxidant enzyme expression. Results derived from real-time quantitative PCR, Western blot, quantitative methylation-specific PCR, and inhibitors assay experiments showed that betaine activated the Nrf2-Keap1-ARE pathway, leading to an increase in the mRNA level of superoxide dismutase, glutathione peroxidase, and heme oxygenase-1. The mechanism may involve betaine's promotion of DNA methyltransferase expression, resulting in increased methylation of Keap1 DNA promoter, which ultimately reduces the level of Keap1 mRNA. Therefore, betaine can directly activate the Nrf2-Keap1-ARE pathway by increasing the methylation level of Keap1 DNA promoter, thereby improving the expression of antioxidant enzymes. This study will contribute to unveiling a new antioxidant mechanism for betaine.
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CHEMICAL ENGINEERING JOURNAL
·
2024
·
IF 13.3
Dechan Lu,Yanping Chen,Longfeng Ke,Zufang Huang,Yudong Lu,Jing Wang
DNA methylation is a promising biomarker for early cancer detection; however, traditional detection methods require bisulfite treatment and nucleic acid amplification, which are costly and impractical for point-of-care use. We propose a facile method that leverages immunoassays and surface-enhanced Raman scattering (SERS) readouts for the multiplex, sensitive, and portable detection of lung cancer-related DNA methylation biomarkers. This approach employs 5-methylcytosine antibody-modified magnetic probes to enrich methylated DNA sequences and uses core–shell nanoparticles with Probe DNA as SERS nanotags for multiplex detection, eliminating the need for bisulfite treatment and nucleic acid amplification. The multiplex SERS immunoassay detects synthetic methylated SHOX2 and RASSF1A with detection limits of 0.52 pM and 0.66 pM, respectively. It shows a strong correlation with quantitative PCR results in analyzing methylated SHOX2 and RASSF1A from cell lines and formalin-fixed paraffin-embedded tissue samples (n = 35). Additionally, the random forest analysis based on methylated SHOX2 and RASSF1A expression distinguishes lung cancer from benign lung diseases with a clinical sensitivity of 96 %, specificity of 90 %, and an AUC value of 0.972. This method offers a significant advancement for DNA methylation research and clinical applications.
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MICROCHIMICA ACTA
·
2024
·Vol. 191
·期 9
·1-9
·
IF 5.3
Qiu Shan,Yang Li,Zhang Xuemei,Zhu Li,Xiong Xiaoli,Xiao Ting,Zhu Liping
An electrochemical aptasensor was developed by utilizing a DNA walker driven by catalytic hairpin assembly (CHA) with kanamycin as the model analyte. Kanamycin bound to the aptamer, causes the release of DNA walker, triggers the CHA reaction, leads to the cyclic movement of the walker’s long arm, and results in cascade amplification of the signal. The guanine-rich sequences of the double-stranded products produced by CHA were folded to form G-quadruplex structures, with electrochemical active molecules Hemin embedded, forms G-quadruplex/Hemin complexes in situ on the electrode surface, thereby achieving sensitive, efficient, and label-free detection of kanamycin with a limit of detection (LOD) of 0.27 pM (S/N = 3). Meaningfully, the aptasensor demonstrated high sensitivity and reliability in the detection of kanamycin in milk and livestock wastewater samples, suggesting that it has great potential for application in detecting antibiotics in food products and water samples from the environment. Graphical abstract
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JOURNAL OF PHYSIOLOGY AND BIOCHEMISTRY
·
2025
·
IF 4.3
Fan Yanan,An Caizhelin,Wang Zhihui,Luo Jia,Wang Wenbin,Luo Qing,Song Guanbin
Hepatocytes are the primary functional cells in the liver, and the malignant transformation of hepatocytes significantly contributes to hepatocellular carcinoma (HCC) progression. Liver fibrosis and cirrhosis caused by extracellular matrix (ECM) remodeling during liver lesions is a pivotal driver of HCC. However, the impact of matrix stiffness on hepatocytes and the underlying molecular mechanisms are not fully understood. Herein, using gelatin/sodium alginate hydrogels with different stiffnesses to simulate the change of matrix stiffness during liver lesions, we found that matrix stiffening leads to a notable decrease in the expression of hepatocyte nuclear factor 4α (HNF4α) and functional hepatocyte genes and a significant increase in the expression of interleukin 6 (IL‒6) in human hepatocyte line L‒02 cells, indicating obvious damage of hepatocyte function. In addition, matrix stiffening causes extensive DNA damage to L‒02 cells. Mechanistically, matrix stiffening upregulates piezo‒type mechanosensitive ion channel component 1 (Piezo1) expression and activates extracellular signal‒regulated kinase 1/2 (ERK1/2) signaling. Piezo1 knockdown suppresses matrix stiffening‒induced functional impairment and DNA damage in L‒02 cells. Moreover, Piezo1 knockdown blocks matrix stiffening‒activated ERK1/2 signaling in L‒02 cells. U0126 (a selective inhibitor of ERK1/2 activation) treatment could rescue matrix stiffening‒induced functional impairment and DNA damage. Taken together, these findings demonstrate that matrix stiffening induces functional impairment and DNA damage in L‒02 cells via the Piezo1‒ERK1/2 signaling pathway, which provides evidence for a better understanding of the hepatocyte function damage caused by tissue mechanical microenvironment change in liver diseases and the mechanotransduction in this process. Graphical abstract An increase in matrix stiffness inhibits HNF4α expression and promotes IL‒6 expression in L‒02 cells. Meanwhile, matrix stiffening induces L‒02 cell DNA damage and upregulates the expression of γ‒H2AX and 53BP1. These findings prove that matrix stiffness is a crucial mechanical regulator of cellular functional impairment and DNA damage and plays an important role in liver disease.
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BIOMATERIALS
·
2024
·
IF 12.8
Jilong Wang,Chenghu Wu,Yiran Wang,Yecheng Shen,Kerui Wu,Yikai Shi,Tianshou Cao,Shanshan Yuan,Yuting Zhu,Yongheng Bai,Jinhai Huang,Yan Zhang,Junjie Deng
Low-expression antigens, especially neoantigens, pose a significant challenge in immunotherapy for low immunogenicity pancreatic cancer. Increasing the tumor mutation burden is crucial to enhance the expression of tumor antigens and improve tumor immunogenicity. However, the incomplete intervention in DNA stability hampers effective elevation of the tumor mutation burden, thus reducing the probability of neoantigen. To address this issue, we have developed a novel nano-regulator that intervenes in the DNA stability of tumor cells, thereby enhancing tumor mutations. This nano-regulator comprises metal-organic frameworks (MOFs) encapsulating DNA damage agent doxorubicin and DNA damage repair inhibitor siRNA-ATR, enabling simultaneous induction of DNA mutations and inhibition of their repair. Importantly, this regulator, named as MOF DOX&siATR , can modulate the tumor gene expression profile, induce the production of neoantigens of Atp8b1, and enhance the immunogenicity of pancreatic cancer. The characteristics of DNA stability intervention by MOF DOX&siATR hold promise for augmenting the immune response in low immunogenic tumors, making it a potential nanomedicine for the treatment of pancreatic cancer.
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CHEMICAL ENGINEERING JOURNAL
·
2022
·
IF 16.744
Xinglu Jiang,Kangjing Li,Beibei Xie,Jun Zhou
Taq DNA polymerase is the most widely used enzyme for PCR amplification and it contains 832 amino acids but no cysteines. Bovine serum albumin (BSA) is an 832-amino acid protein containing 35 cysteines, and at high temperature BSA can produce protein hydrogels through recombined disulfides and noncovalent bonds. When hydrogels are prepared using a mixture of Taq DNA polymerase and BSA, no disulfide bonds occur between them, and noncovalent bonds are the main forces holding together the Taq DNA polymerase with the hydrogels. During PCR amplification, temperature cycling can destroy the noncovalent bonds, thereby releasing Taq DNA polymerase from the hydrogels to synthesize DNA. The release rate, regulated through the variation of BSA concentration, allows for velocity-controlled amplification of DNA. As the amount of DNA products exceeds that of the Taq DNA polymerase released from hydrogels, a linear amplification replaces the exponential PCR amplification. When the amplification velocity begins to decrease, the time needed for reaching a plateau increase. This provides a broader detection window for the analysis of the end products, which correspond to the difference compared with the initial DNA concentration. The number of primers is continuously reduced with increasing cycles, which further causes a decline in the DNA amplification efficiency. The difference in the initial DNA template may cause a different consumption of primers, which can cause errors in the Ct values in the case of qPCR. Based on the slopes of the linear amplification curve, the Ct values can be calibrated to reduce this error, and a more accurate detection of the initial content of the target DNA can be implemented.
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JOURNAL OF PHYSICS D-APPLIED PHYSICS
·
2025
·Vol. 58
·期 13
·135207
·
IF 3.2
Liu Jinren,Wang Xiangni,Cui Yixin,Xu Guimin,Lu Jiajia,He Zhirou,Xu Yulin,Li Rongrong,Gao Lingge,Wang Hong,Gao Junhong,Wu Xili,Shi Xingmin,Zhang Guanjun
Cisplatin resistance coupled with adverse effects presents substantial hurdles in the treatment of ovarian cancer. Both cold atmospheric plasma and plasma-activated liquid (PAL) hold the ability to induce DNA damage, which is a primary target of cisplatin. In this study, we investigate the potential of PAL in augmenting cisplatin sensitivity or diminishing the necessary drug dosage by impairing the DNA of cisplatin-resistant cancer cells. We found that plasma irradiated medium exceeding 40 s could decrease the viability, induce DNA damage, trigger cell cycle arrest, and augment apoptosis and autophagy in two types of cisplatin-resistant ovarian cancer cells (A2780/DDP and SKOV3/DDP) after a 24 h incubation period. Notably, pre-treatment with 40 s plasma-activated medium (PAM) for 2 h significantly enhanced the anti-cancer effect of cisplatin administered at lower doses on these cell types. RNA-seq, quantitative real-time PCR and Western blotting analysis revealed that PAM treatment could downregulate the expression of several pivotal proteins involved in nucleotide excision repair (NER) and homologous recombination repair (HRR). The mouse xenograft tumor model demonstrated that plasma-activated saline and cisplatin synergistically inhibited tumor growth and significantly suppressed the expression of proteins related to HRR and NER pathways. We identified plasma-derived peroxide generation in PAM as crucial in these processes. These results pave the way for considering PAM as an adjuvant treatment for cisplatin-resistant ovarian cancer and other abdominal cancers. Additionally, the findings indicate the feasibility of reducing cisplatin dosage to alleviate side effects through sensitization.
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MICROCHEMICAL JOURNAL
·
2024
·
IF 4.9
Rikun Cui,Linna Li,Lili Liu,Zhen Li,Zhenzhao Wang,Xingtong Liu,Lei Tian,Fujun Yao,Xiaofeng Kang,Yanli Guo
Nucleic acid aptamers, which are short single-stranded RNA or DNA molecules, undergo conformational changes upon recognizing small molecules. However, their structural characterization is challenging due to the flexibility of single-stranded oligonucleotides. Nanopores provide a sensitive method for single-molecule analysis of molecular conformational changes as they transport through the pore. In this study, we employed α-hemolysin (α-HL) nanopore with high spatiotemporal resolution to investigate the conformational changes of DNA aptamer (ADO-23) upon binding to adenosine (Ado) molecules. Unlike the events generated by the aptamer alone, the DNA-Ado complex produced two distinct new characteristic events, indicating the formation of two unique secondary structures. By analyzing current events, the structures formed by the DNA-Ado complex were speculated to be hairpin DNA and G-quadruplex (G4) DNA. Furthermore, we demonstrated the potential of the aptamer as a chiral selector within the nanopore. The results showed that the aptamer specifically bound to D-Ado and not to L-Ado. Finally, we realized specific and sensitive detection of small molecule Ado by α-HL pore, with a detection limit as low as 10.5 nM, and successfully applied it to the detection of real human serum samples. The nanopore platform provides a powerful tool for studying small molecule-biomolecule conformational changes, and we constructed sensors for detecting adenosine through the aptamer. This work also offers a promising new approach to the recognition of enantiomers at the single-molecule level.
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Chemosensors
·
2025
·Vol. 13
·期 2
·32
·
IF 3.7
Yang Dai,Xiuran Mao,Maimaiti A. Abulaiti,Qianyu Wang,Zhihao Bai,Yifeng Ding,Shuangcan Zhai,Yang Pan,Yue Zhang
Monitoring of immune factors, including interferon-gamma (IFN-γ), holds great importance for understanding immune responses and disease diagnosis. Wearable sensors enable continuous and non-invasive detection of immune markers in sweat, drawing significant attention to their potential in real-time health monitoring and personalized medicine. Among these, electrochemical sensors are particularly advantageous, due to their excellent signal responsiveness, cost-effectiveness, miniaturization, and broad applicability, making them ideal for constructing wearable sweat sensors. In this study, we present a flexible and sensitive wearable platform for the detection of IFN-γ, utilizing a DNA hydrogel with favorable loading performance and sample collection capability, and the application of mobile software achieves immediate data analysis and processing. This platform integrates three-dimensional DNA hydrogel functionalized with IFN-γ-specific aptamers for precise target recognition and efficient sweat collection. Signal amplification is achieved through target-triggered catalytic hairpin assembly (CHA), with DNA hairpins remarkably enhancing sensitivity. Ferrocene-labeled reporting strands immobilized on a screen-printed carbon electrode are displayed via CHA-mediated strand displacement, leading to a measurable reduction in electrical signals. These changes are transmitted to a custom-developed mobile application via a portable electrochemical workstation for real-time data analysis and recording. This wearable sensor platform combines the specificity of DNA aptamers, advanced signal amplification, and the convenience of mobile data processing. It offers a high-sensitivity approach to detecting low-abundance targets in sweat, paving the way for new applications in point-of-care diagnostics and wearable health monitoring.
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Materials Today Bio
·
2024
·
IF 8.7
Hongjian Liao,Yuchao Cao,Can Hu,Shangfeng Shen,Zhifei Zhang,Dairong Li,Yonghong Du
Lung cancer is the deadliest kind of cancer in the world, and the hypoxic tumor microenvironment can significantly lower the sensitivity of chemotherapeutic drugs and limit the efficacy of different therapeutic approaches. In order to overcome these problems, we have designed a drug-loaded targeted DNA nanoflowers encoding AS1411 aptamer and encapsulating chemotherapeutic drug doxorubicin and oxygen-producing drug horseradish peroxidase (DOX/HRP-DFs). These nanoflowers can release drugs in response to acidic tumor microenvironment and alleviate tumor tissue hypoxia, enhancing the therapeutic effects of chemotherapy synergistic with sonodynamic therapy. Owing to the encoded drug-loading sequence, the doxorubicin loading rate of DNA nanoflowers reached 73.24 ± 3.45%, and the drug could be released quickly by disintegrating in an acidic environment. Furthermore, the AS1411 aptamer endowed DNA nanoflowers with exceptional tumor targeting properties, which increased the concentration of chemotherapeutic drug doxorubicin in tumor cells. It is noteworthy that both in vitro and in vivo experiments demonstrated DNA nanoflowers could considerably improve the hypoxia of tumor cells, which enabled the generation of sufficient reactive oxygen species in combination with ultrasound, significantly enhancing the therapeutic effect of sonodynamic therapy and evidently inhibiting tumor growth and metastasis. Overall, this DNA nanoflowers delivery system offers a promising approach for treating lung cancer.
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