ANALYTICA CHIMICA ACTA
·
2021
·
IF 6.558
Xiangyu Ma,Kun Qian,Onome Ejeromedoghene,Martha Kandawa-Schulz,Wei Song,Yihong Wang
In this work, we report AuNPs-decorated pyrolyzed Co-BDC nanosheets (p-Co-BDC/AuNPs) as high-performance electrocatalyst for developing an electrochemical platform. p-Co-BDC/AuNPs as a new electrocatalyst showed superior electrocatalytic activity towards the electrochemical oxidation of methylene blue (MB). Besides, magnetic p-Co-BDC/AuNPs can be well immobilized on the magnetic glassy carbon electrode without further assistance. The oxidation of MB can be reduced by ascorbic acid . Inspired by this phenomenon, an electrochemical biosensor was constructed based on multiple signal amplification for the diagnosis of miRNAs . Firstly, p-Co-BDC/AuNPs enhanced the electrochemical oxidation of MB. Then, strand displacement amplification reaction can form lots of double helix structure DNA to embed more MB molecules. Finally, ascorbic acid in the electrolyte was utilized to reduce the oxidation of MB and improve the electrochemical signal of MB electro-oxidation. The linear detection range for the detection of miRNAs is 100 aM to 10 nM, and the limit of detection is 86 aM. Furthermore, the constructed biosensor also displayed satisfactory selectivity , good reproducibility, and excellent recovery in the detection of real samples. We are convinced that our proposed multiple signal amplification strategy will provide more promising methods for the diagnosis of cancer.
研究领域:
生物化学
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分子生物学
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分析化学
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PubMed
高影响因子
ACS Sensors
·
2026
·
IF 10.9
Yinghui Sun,Rui Wang,Hongxiao Sun,Jiadong Chen,Chengwei Liang,Zhiyang Zhang,Yujie Feng,Lingxin Chen,Xiaokun Wang
MicroRNAs (miRNAs) are emerging biomarkers for early hepatocellular carcinoma (HCC) detection, but most CRISPR−Cas12a assays rely on reverse transcription and preamplification and often lack simple visual readouts. Here, we develop a reverse transcription-free, cleavage-guided strategy that converts miRNA recognition into visual outputs. In the presence of the target miRNA, a scaffold RNA forms an active crRNA that triggers Cas12a trans-cleavage using a preformed dsDNA activator, enabling two readout formats. In a homogeneous non-crosslinking colorimetric assay (mC-NCA), Cas12a cleavage regulates π−π-stacking-mediated gold nanoparticle (AuNP) aggregation to produce a rapid visual colorimetric response. In a heterogeneous versatile lateral flow assay (mC-vLFA), the cleavage of partially hybridized DNA−magnetic bead probes generates a target-dependent test line while maintaining a built-in control line. Using miRNA-21 and miRNA-122 as targets, mC-NCA shows calculated limits of detection (LODs) of 1.62 and 1.64 fM with a linear range of 50 fM−500 pM, whereas mC-vLFA shows calculated LODs of 1.59 and 1.40 fM with a linear range of 10 fM−10 nM. In a preliminary clinical evaluation, both formats show good agreement with reverse-transcription quantitative polymerase chain reaction (RT-qPCR) and clinical assessment while enabling faster detection with minimal instrumentation. Overall, this dual-readout platform couples RT-free CRISPR−Cas12a miRNA recognition with cleavage-directed visual signal transduction, enabling rapid and low-instrument-dependence CRISPR diagnostics for miRNAs.
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PubMed
高影响因子
TALANTA
·
2024
·
IF 5.6
Ruili Wang,Ruiying Peng,Liran Song,Jishan Li
Herein, we present a colorimetric sensing strategy for the identification and quantification of tumor-associated miRNAs based on dual DNAzyme amplification. In this sensing ensemble, the substrate portion of the Pb 2+ -dependent 8−17 DNAzyme combines with the G-quadruplex portion to form a hairpin substrate strand. The two split 8−17 DNAzyme strands are partially complementary to the substrate strand and serve as a recognition unit for binding the target miRNA. In the presence of the target miRNA, the activated DNAzyme cleaves the substrate strand, releasing the G-quadruplex. This G-quadruplex binds to hemin to form a G-quadruplex/hemin complex with horseradish peroxidase (HRP)-like properties, which catalyzes the oxidation of ABTS 2− by H 2 O 2 . This oxidation reaction produces a colorimetric signal output, enabling the detection of the target miRNA. Under the optimal reaction conditions explored in this study, the constructed sensing ensembles tailored for each of the specific target miRNAs successfully identified and quantified the four target miRNAs—miR-122, miR-21, miR-335, and miR-155—in both buffer solutions and cell extracts. This colorimetric sensing strategy offers significant advantages in terms of simplicity, cost, and versatility and holds great potential for wide application in biomedical research and clinical diagnostics.
研究领域:
生物化学
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分子诊断学
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分析化学
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PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2026
·
IF 7.3
Zhang-Feng Tang,Jun Zeng,Yao Chen,De-Huan Yang,Jing-Hao Fu,Zeng-Ping Chen,Ru-Qin Yu
MicroRNAs (miRNAs) have emerged as potential biomarkers for human diseases. Classification models established on the expression levels of multiple miRNAs can be used to realize a reliable diagnosis of diseases. The detection of expression patterns of multiple miRNAs using traditional methods is generally cost-prohibitive, labor-intensive, and limited by the requirements of sophisticated facilities. DNA molecular computing technology attempts to implement a multiple-miRNA-based disease classification model through reactions among DNA probes and miRNAs without the need to quantify each individual miRNA, and hence has the potential to realize fast and cheap clinical diagnosis of early-stage diseases. However, existing DNA molecular computing frameworks for disease diagnosis struggle in faithfully implementing even linear classification models with nonzero classification thresholds, which hinders their practical clinical applications. Herein, for the first time, a novel DNA molecular computing platform enabling full and accurate implementation of multiple-miRNA-based disease classification models is proposed. It consists of “multiplication,” “addition,” “calibration,” and “diagnosis” modules. The first two modules are implemented experimentally through “one-pot” reactions among DNA probes and miRNAs, while the last two modules are carried out in silico. Experimental results have demonstrated that such a hybrid implementation strategy breaks through the bottleneck of conventional DNA molecular computing frameworks for disease diagnosis, and faithfully implements a disease classifier for discriminating diseased samples from normal ones. The new platform has the advantages of reduced design complexity, easy experimental implementation, and robustness to variations in signal amplification factors across samples, and opens up an avenue for point-of-care clinical diagnosis of early-stage diseases.
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高影响因子
Analytical Methods
·
2023
·Vol. 15
·期 24
·2915-2924
·
IF 3.1
Wei Xu,Yu Zhang,Dianhai Hou,Jianjun Shen,Jinhua Dong,Zhiqin Gao,Honglin Liu
Simultaneous and ultrasensitive detection of multiple microRNA (miRNA) biomarkers is an essential precondition for early cancer diagnosis and treatment. Here we developed a sandwich surface-enhanced Raman scattering (SERS) sensor based on Au@Ag core–shell nanorods combined with duplex specific nuclease-mediated signal amplification (DSNSA) for quantitative detection of multiple breast cancer miRNA biomarkers. The DSNSA strategy enables quantitative detection of target miRNA through rehybridizing the capture probe DNA-SERSnanotag conjugates to trigger signal amplification. The Au@Ag core–shell nanorods coated with an Ag shell exhibit excellent SERS performance, implying that molecules can be concentrated by the Ag shell at the hot spots. By monitoring the Raman signal attenuation of hot spots in the presence of target miRNAs, three breast cancer associated miRNAs (miR-21, miR-155, and let 7b) were simultaneously determined using the sandwich SERS sensor, and their detection limits (LODs) were 0.05 fM, 0.063 fM and 0.037 fM, respectively. These results indicated that our sandwich SERS sensor combined with the DSNSA strategy holds remarkable promise for multiplex detection of cancer biomarkers and contributes to early diagnosis of cancer.
研究领域:
生物医学
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分子诊断
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纳米技术
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DOI
PubMed
高影响因子
BIOMEDICAL MICRODEVICES
·
2026
·Vol. 28
·期 1
·3
·
IF 3.3
Yang Chun-Guang,Han Yi,Zhang Jing-Jing,Xu Li-Wen,Li Jin
The expression levels of miRNAs in cells or biological fluids are closely associated with the onset and progression of many diseases, making the detection of miRNAs in serum and plasma increasingly important. However, due to the low levels of miRNAs in biological fluids and the complexity of their surrounding environment, their analysis faces challenges related to accuracy and reproducibility. We developed a duplex-specific nuclease (DSN)-mediated detection method for the direct detection of miRNA-21 in real samples. This method eliminates the need for total RNA extraction from real samples using commercial kits, a key advantage over traditional methods requiring pre-treatment. As verified by performance evaluation, this method exhibits a linear range of 5.00 fmol·L − 1 to 500 pmol·L − 1 and a limit of detection (LOD) of 2.66 fmol·L − 1 for miRNA-21. Direct detection of miRNA-21 in real samples yielded favorable results. These findings confirm that the method holds great application potential for the direct detection of miRNA-21 in real biological samples and exhibits promising prospects in clinical cancer diagnosis and drug screening.
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高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2025
·
IF 8.5
Hui Li,Zongshan Zhang,Wantong Hao,Huixin Shi,Ruiyang Yin,Jing Xu
MicroRNAs (miRNAs) are key regulators of gene expression and play a crucial role in various biological processes. However, existing methods for miRNA detection face challenges in terms of sensitivity, specificity and ease of use. These limitations highlight the need for innovative and highly sensitive detection platforms capable of accurately quantifying miRNAs. In this work, an integrated biochip based on a self-powered sensor system developed for the ultra-sensitive detection of miRNAs-21 and miRNA-20a. The system uses a bilirubin oxidase (BOD)-supported bio-cathode and a glucose oxidase (GOD)-supported bio-anode to form a closed loop that converts chemical energy into electrical energy. The complementary chains of target miRNAs are modified on the bio-anode using a bio-conjugate amplification strategy, enabling the system to generate an electrical signal proportional to the concentration of miRNAs. The sensor has a wide linear range of 5 fmol/L to 10 nmol/L for miRNA-20a and 1 fmol/L to 10 fmol/L for miRNA-21, with low detection limits of 0.176 fmol/L and 0.138 fmol/L, respectively. These detection limits are significantly lower than those achieved by existing technologies, making our biosensor a significant advancement in the field. In addition, the sensor system is integrated into a portable chip that transmits electrical signals in real time via Bluetooth to a mobile phone interface, enabling convenient and rapid analysis.
研究领域:
生物医学工程
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分子生物学
·
分析化学
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PubMed
高影响因子
Polymer Chemistry
·
2017
·Vol. 8
·期 27
·4043-4051
·
IF 5.375
Wenhua Yang,Changmin Yu,Chunxian Wu,Shao Q. Yao,Shuizhu Wu
MicroRNAs (miRNAs) are small regulatory noncoding RNAs that control a variety of biological processes. The regulation of many endogenous miRNAs is tightly associated with various diseases. Thus, the design of miRNA delivery systems with minimal endolysosomal trapping, efficient delivery and controlled miRNA release is urgently needed. Herein, we have developed a new star polymer which consists of a β-cyclodextrin (βCD) core and multiple cell-penetrating poly(disulfide) (CPD) arms. By subsequently loading the system with miRNAs and small molecule drugs, we have successfully proven that this novel drug delivery platform could efficiently enter mammalian cells (<2 h) without apparent endolysosomal trapping. The global Pearson's R value was calculated to be 0.31 between our complex and endolysosome, which is far below the threshold of >0.5 required for correlation. In addition, the GSH-triggered degradation of CPD arms and the subsequent intracellular release of miR-203 and CPT, as well as the combination therapeutic effects have been successfully demonstrated. In this way, we show that this novel platform could be used in future to minimize potential cytotoxicity encountered by many existing cationic branched polymer systems in miRNA delivery. Our results provide important starting points for using CPD-based polymers to design personalized delivery platforms.
研究领域:
药学
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生物技术
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分子生物学
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药物递送系统
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DOI
高影响因子
Frontiers in Genetics
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2021
·
IF 4.599
Zhang Jing,Gao Juan,Lin Dengna,Xiong Jing,Wang Jialei,Chen Junfeng,Lin Bingliang,Gao Zhiliang
Acute-on-chronic liver failure (ACLF) is a severe syndrome associated with high mortality. Alterations in the liver microenvironment are one of the vital causes of immune damage and liver dysfunction. Human bone marrow mesenchymal stem cells (hBMSCs) have been reported to alleviate liver injury via exosome-mediated signaling; of note, miRNAs are one of the most important cargoes in exosomes. Importantly, the miRNAs within exosomes in the hepatic microenvironment may mediate the mesenchymal stem cell (MSC)-derived regulation of liver function. This study investigated the hepatocyte exosomal miRNAs which are regulated by MSCs and the target genes which have potential in the treatment of liver failure. Briefly, ACLF was induced in mice using carbon tetrachloride and primary hepatocytes were isolated and co-cultured (or not) with MSCs under serum-free conditions. Exosomes were then collected, and the expression of exosomal miRNAs was assessed using next-generation sequencing; a comparison was performed between liver cells from healthy versus ACLF animals. Additionally, to identify the intracellular targets of exosomal miRNAs in humans, we focused on previously published data, i.e., microarray data and mass spectrometry data in liver samples from ACLF patients. The biological functions and signaling pathways associated with differentially expressed genes were predicted using gene ontology and Kyoto Encyclopedia of Genes and Genomics enrichment analyses; hub genes were also screened based on pathway analysis and the prediction of protein-protein interaction networks. Finally, we constructed the hub gene-miRNA network and performed correlation analysis and qPCR validation. Importantly, our data revealed that MSCs could regulate the miRNA content within exosomes in the hepatic microenvironment. MiR-20a-5p was down-regulated in ACLF hepatocytes and their exosomes, while the levels of chemokine C-X-C Motif Chemokine Ligand 8 (CXCL8; interleukin 8) were increased in hepatocytes. Importantly, co-culture with hBMSCs resulted in up-regulated expression of miR-20a-5p in exosomes and hepatocytes, and down-regulated expression of CXCL8 in hepatocytes. Altogether, our data suggest that the exosomal miR-20a-5p/intracellular CXCL8 axis may play an important role in the reduction of liver inflammation in ACLF in the context of MSC-based therapies and highlights CXCL8 as a potential target for alleviating liver injury.
研究领域:
肝病学
·
干细胞生物学
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分子生物学
·
基因组学
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PubMed
高影响因子
RSC Advances
·
2026
·Vol. 16
·期 8
·6844-6855
·
IF 6.1
Xinqin He,Mengjie Yang,Xia Zhang,Bin Qiu,Hong Jiang
Early detection of endometrial cancer remains challenging because of the lack of sensitive and specific diagnostic methods. This study develops a near-infrared (NIR)-activatable sensing platform with an AND logic gate to simultaneously detect miR-21 and miR-155, two promising endometrial cancer biomarkers. The sensing platform integrates acid-sensitive zeolitic imidazolate framework-8 (ZIF-8)-coated upconversion nanoparticles (UCNP@ZIF-8) with ternary DNA strands, consisting of a BHQ2-modified long strand (S2), a Cy5-labelled short strand (S3), and a photocleavable switch-bearing strand (S1). In a weakly acidic environment, the ZIF-8 shell decomposes to release UCNPs and DNA strands. Under 808 nm excitation, the UCNPs convert NIR light to ultraviolet or visible emission, cleaving the photocleavable linkers on S1 and exposing the toehold domain. The target miRNAs subsequently initiate a toehold-mediated strand displacement reaction, simultaneously displacing S1 and S3, thereby restoring Cy5 fluorescence. The biosensor demonstrated excellent sensitivity, with detection limits of 0.28 nM for miR-21 and 0.35 nM for miR-155. Validation in serum samples revealed recovery rates of 92–103.08%, confirming its potential for clinical application in complex biological environments.
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CHEMICAL ENGINEERING JOURNAL
·
2026
·
IF 12.5
Chenchen Guo,Nan Ye,Huimin Xiao,Yu Miao,Zhihua Xiao,Qin Liu,Yuhong Ren
Self-healing hydrogel formed via synergistic chemical and physical crosslinking. • Effective loading and sustained release of proteins and nucleic acids. • Multifunctional hydrogel with antioxidant and antibacterial activities. • Synergistic therapeutic effects for rapid diabetic wound repair. Diabetic wound healing is a protracted process that presents a significant clinical challenge due to impaired angiogenesis and chronic inflammation. While multifunctional hydrogels offer a promising strategy for effective wound management, achieving synergistic tissue regeneration remains a critical hurdle. In this study, an in situ forming, injectable composite hydrogel comprising methacrylated chitosan (CSMA) and dopamine-modified hyaluronic acid (HADA), crosslinked via a combination of Schiff base formation, Michael addition, and physical interactions, was developed. This platform is engineered to co-load and effectively release recombinant laminin fragment α5E8 (LM-α5E8) and pro-healing microRNAs for the synergistic acceleration of diabetic wound repair. The resulting CMHD@α5E8/miRNAs hydrogel exhibits excellent mechanical properties, intrinsic antibacterial activity, and potent antioxidant capacity, alongside confirmed in vitro hemocompatibility and cell viability, while effectively suppressing pro-inflammatory responses. In vivo evaluations using a full-thickness diabetic murine skin defect model demonstrated that this hydrogel significantly expedites wound closure. Furthermore, the delivered miRNAs optimize the wound microenvironment by silencing specific negative regulators, thereby activating essential signaling pathways to drive cellular migration and angiogenesis. Consequently, comprehensive histological and molecular validations corroborated the accelerated angiogenesis and enhanced collagen network reconstruction. Overall, this multifunctional hydrogel represents a highly effective wound dressing, offering substantial therapeutic potential for diabetic skin injury management. Download: Download high-res image (186KB) Download: Download full-size image
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INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2024
·
IF 7.7
Yingxue Li,Tingrui Zhang,Gang Bai,Mengchun Chen,Xiaodong Lei,Li Ye,Hua Yu,Zhichao Fan,Tao Yu
MicroRNAs (miRNAs) play an important role in the process of heart failure (HF) and are emerging biomarkers that can be used for the auxiliary diagnosis of HF. However, it is very challenging to accurately analyze the expression levels of trace miRNAs in complex clinical samples. Here, we developed an enzyme-free colorimetric sensor for the ultrasensitive detection of miRNA-423-5p (HF-associated miRNA) based on three-dimensional DNA walkers constructed from functional nucleic acids and gold nanoparticles (AuNPs). DNAzyme with cleavage activity was specifically activated by miRNA-423-5p to sustainably cleave the substrate, thereby releasing the trigger sequence to initiate the subsequent mismatched catalytic hairpin assembly (MCHA) cycle. Then, as the MCHA cycle proceeded to continuously expose the G-quadruplex (GQ) sequence, the sequence bound with hemin to form a large amount of GQ/hemin DNAzyme on the surface of the AuNPs, which rapidly catalyzed the chromogenic oxidation of 3,3′,5,5′-tetramethylbenzidine to yield an amplified colorimetric signal readout. The colorimetric sensor exhibited an ultralow detection limit (32 fM), showed excellent specificity and performed well in serum samples. The sensor was applied to detect miRNA-423-5p in clinical plasma samples from healthy individuals and HF patients, and the results revealed its good clinical application in HF diagnosis. Thus, the developed colorimetric sensor provides a convenient detection tool for early screening and diagnosis of HF, as well as for pathophysiological studies.
研究领域:
生物标志物检测
·
纳米技术
·
分子诊断
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心血管疾病
·
生物医学工程
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PubMed
高影响因子
危险品化学品经营许可证（带存储）