查找使用阿拉丁产品的科研文献

Detection of microRNAs by the combination of Exonuclease-III assisted target recycling amplification and repeated-fishing strategy

Xiao-Mei Yan,Yin-Qi Wang,Yao Chen,Zeng-Ping Chen,Ru-Qin Yu

A simple but effective method for the detection of miRNAs was proposed by integrating exonuclease-III assisted target recycling amplification and repeated-fishing strategy. In the proposed method, exonuclease-III assisted target recycling amplification reaction is adopted to produce a large amount of DNA fragments with fluorescence group at its 5′ end in the presence of the target miRNA, which are then repeatedly fished out from the reaction mixture by a gold foil modified with a capture probe and transferred into a so-called ‘product tube’. The amount of the target miRNA can then be determined from the fluorescence measurement of the solution in the ‘product tube’. Application to the detection of miRNA-155 in samples of KH-2 and BRSA-2B cells revealed that the proposed method could achieve sensitive and accurate quantification of the target miRNA with a limit of detection of 36 fM and recovery rates in the range from 96.2% to 105%. Its simplicity, sensitivity and resistance to possible fluorescence interferences in complex biological samples make the proposed method a potentially competitive alternative for miRNAs detection in complex biological samples.

A simple and efficient strategy for trace detection of ferroptosis-related miRNAs based on novel hydrophobic paper-based plasmonic substrate and “inverse molecular sentinel (iMS)” nanoprobes

Wang Youwei,Chen Bing,Fan Jiang,Wang Zhong

Monitoring ferroptosis-related miRNAs is crucial for the treatment and prognosis of patients with intracerebral hemorrhage. In this work, a novel hydrophobic paper (h-paper)-based plasmonic substrate was produced by dropping DS Au nanorods with a narrow range of sizes and morphologies onto h-paper. Raman reporter molecules were adsorbed to the array surface, and surface-enhanced Raman scattering spectra at randomly selected points reveal uniform and significant SERS enhancement. Hairpin DNAs labelled with Raman reporters and hybridized with placeholder DNAs were decorated on SERS substrate to fabricate SERS biosensor. Target miRNAs initiated the “inverse Molecular Sentinel” process. During the process, PHs were removed and the conformation of HPs changed toward the hairpin structure, thus eliciting the proximity of Raman reporter to substrate and a stronger SERS signal. The proposed SERS biosensor performs well in terms of stability, reproducibility, and selectivity. The limits of detection of miR-122-5p and miR-140-5p in serum were 4.17 aM and 4.49 aM, respectively. Finally, the fabricated SERS biosensor was applied to detect miR-122-5p and miR-140-5p in ICH patients and healthy subjects, and the results obtained by SERS were consistent with the results from quantitative real-time polymerase chain reaction, revealing the accuracy of the method. This simple, rapid approach offers great potential for the simultaneous detection of miRNAs in practical clinical applications.

Functionalized nanopores based on hybridization chain reaction: Fabrication and microRNA sensing

Xia Qiu,Jingyi Dong,Qingshan Dai,Mimi Huang,Yongxin Li

Enzyme-free hybridization chain reaction (HCR) technology is often used as a signal amplification tool for the detection of different targets. In this study, an ultrasensitive and label-free method for detecting miRNA-21 was developed using the nanopore ionic current rectification (ICR) technology coupled with HCR technology. The probe oligonucleotide (DNA1) was combined with the gold-coated nanopore through the Au–S bond to form a DNA1-functionalized gold-coated nanopore (DNA1-Au-coated nanopore). Since miRNA-21 is partially complementary to DNA1, it can be selectively recognized by DNA1-functionalized gold-coated nanopores. The target (miRNA-21) can induce the opening of hairpin DNA and HCR reaction after the introduction of hairpin DNA H1 and H2. The concentration of miRNA-21 will affect the combination of H1 and H2 on the inner wall of the nanopore, and its surface charge will change with the internal modification, thereby changing the ion current rectification ratio . Under the condition that the concentration of H1, H2 and HCR reaction time are constant, the change of ICR ratio is linearly correlated with the logarithm of miRNA-21 concentration within a certain range, which shows that the sensing strategy we designed can achieve target miRNA-21 detection. This ultrasensitive miRNA holds great promise in the field of cancer diagnosis.

Ultrasensitive electrochemiluminescence biosensor constructed by luminol-diazonium ion functionalized Au/MXene nanocomposites for early stage detection of disease in human

Li Tan,Wenyi Zhang,Ziqi Lian,Yu Wang,Wenjing Gao,Tao Zeng,Yun Chen,Meng Yu,Benlin Hong,Rongkai Ye,Ying Ma,Yuchan Liu,Jianqiang Hu,Jiaju Mo,Aiqing Li

ABSTRACT MicroRNAs (miRNAs) are important biomarkers for tracking occurrence and development of many diseases, therefore, the reliable, sensitive, fast and accurate detection of miRNAs plays an extremely critical role in clinic diagnosis and treatment of early stage diseases. Herein, we developed ultrasensitive electrochemiluminescence (ECL) biosensor constructed by luminol-diazonium ion functionalized Au/MXene nanocomposites for early stage detection of kidney disease in human. The synergistic effect of MXene and Au nanoparticles significantly enhanced the ECL emission of luminol up to 6.9-fold due to their shortened electron transfer distance and excellent catalytic performance. The loading efficiency of luminol on MXene could be as high as ∼7%, enabling the enrichment of luminol on MXene nanosheets. The enhanced ECL signal and excessive loading of luminol allowed the Au-MXene-luminol as an excellent ECL labeling agent, which was further developed for the determination of miRNA-377 according to the sandwich-like assay. The as-developed miRNA-377 biosensor had ultrahigh sensitivity, wide linear range (from ∼50 aM to ∼1 nM), low limit of detection (∼13.2 aM) and high selectivity (∼82.6%) for highly homologous miRNA with mismatched single-bases. The ultrasensitive ECL biosensor was successfully applied to determine miRNA-377 in the serum samples of early stage kidney patients, which were well consistent with the clinic standard of quantitative real-time polymerase chain reaction method. The ultrasensitive ECL-labeling biosensor with high selectivity is very promising for universally screening any kinds of biomarkers with corresponding specific targets.

RNA-Modified Graphene Oxide-Encapsulated Gold Nanoparticles-Based Stable Fiber Optic MiRNA-126 Sensor

Bing Xu,Liu Yang,Liyun Ding,Zhihui Luo,Zhihong Pan

Time-course miRNA alterations and SIRT1 inhibition triggered by adolescent lead exposure in mice

Liu Rundong,Wang Yawei,Bai Lin,Wang Ruike,Wu Yingying,Liu Mengchen,Li Qiong,Ba Yue,Zhang Huizhen,Zhou Guoyu,Cheng Xuemin,Huang Hui

Sirtuin 1 (SIRT1), the NAD-dependent histone deacetylase, has been extensively investigated due to its cognitive protective effect. Studies suggest microRNAs (miRNAs) and histone modifications are key epigenetic regulators of gene expression and play important role in brain development. We previously showed that cognitive impairment by lead (Pb) was associated with downregulation of SIRT1, but the epigenetic role of this is unclear. Thus, we exposed 4-week-old male mice to 0.2% lead acetate solution for three months, and subsequently extracted brain homogenate from mice cortex and hippocampus at the age of 1, 4, and 16 months, respectively. In this study, we found that the protein level of SIRT1 was inhibited in the hippocampus and cortex of 16-month-old aged mice exposed to Pb. Moreover, changes in the levels of miR-138-5p and miR-141-3p, which were considered to the mechanistic target of SIRT1 by bioinformatic analysis, were negative correlations SIRT1 protein expression. We also found miR-34c-3p expression was increased in the cortex of mice at the age of 16 months. Collectively, our results showed the expression of neural SIRT1 and three selected microRNAs at different age nodes of mice for the first time of following Pb exposure. Our results suggest that additional efforts should focus on the consequences of early Pb exposure from an epigenetic perspective.

Unraveling Bone-Skin Crosstalk Enables miRNA Nanoformulation for Cutaneous Neurovascular Reconstruction in Diabetic Mice

Tao Shen,Hongdou Shen,Weiyue Wu,Zhuoying Jiang,Bin Liu,Yansi Xian,Rui Peng,An Yan,Yu Ben,Yong Shi,Xiang Chen,Baosheng Guo,Qing Jiang,Weijun Wang,Pingqiang Cai

The clinical intractability of diabetic foot ulcers stems from a profound uncoupling of cutaneous neurovascular networks, rendering standard metabolic and topical interventions largely palliative. Paradoxically, remote orthopedic trauma robustly accelerates distal skin repair, yet the systemic molecular mediators driving this “bone-skin crosstalk” remain undefined, precluding its translation into non-invasive therapies. Here, we establish that macroscopic bone fracture expedites diabetic wound healing through the systemic release of exosomal miR-130b-3p, a potent orchestrator of coupled angiogenesis and neurogenesis. To recapitulate this physiological axis non-invasively, we engineered a self-assembling, cholesterol-modified agomir-130b-3p nanocomplex that could bypass endolysosomal degradation. For sustained spatial delivery, these carrier-free nanoassemblies were incorporated into an in situ photocrosslinkable methacrylated collagen/silk fibroin hydrogel, creating a bio-instructive extracellular matrix that prolongs microRNA bioavailability. In streptozotocin-induced diabetic mice, hydrogel-mediated agomir delivery achieved 97.2% full-thickness wound closure. Advanced volumetric light-sheet imaging of chemically cleared whole-mount skin confirmed the robust spatiotemporal reconstruction of deep vascular and neural networks. These findings decode a distinct exosome-mediated inter-organ repair mechanism and demonstrate that biomimetic microRNA nanoformulations can effectively translate systemic physiological cues into localized, high-efficacy therapeutics for ischemic neuropathic wounds.

Logic-gated trimodal CRISPR/Cas12a biosensor for autoimmune hepatitis monitoring via dual miRNA profiling

Xiaolong Li,Daxiu Li,Yanni Wang,Caiyue Xiang,Shuang Tan,Rou Fan,Kai Shi,Wenjiao Zhou

A novel logic-gated D -CHA/Cas12a biosensor enables accurate dual-miRNA detection. • PS-G-triplex mediates label-free fluorescence, colorimetric, and RGB outputs. • Ultra-low femtomolar sensitivity was realized through D -CHA and Cas12a amplification. • Dynamic AIH progression and prednisone efficacy in mice were tracked successfully. An innovative trimodal optical biosensor incorporating dual catalytic hairpin assembly ( D -CHA) and CRISPR/Cas12a was fabricated to specifically and sensitively detect miR-21 and miR-122, two core biomarkers of autoimmune hepatitis (AIH). The platform employs a phosphorothioate (PS)-modified G-rich hairpin (SHG3) as the reporter, enabling fluorescence, colorimetric, and smartphone-based Red Green Blue (RGB) signal readouts. Target-driven AIH produces extended double-stranded DNAs that trigger Cas12a trans-cleavage, thereby cleaving SHG3 to release G-rich strands for PS G-triplex (SG3) formation. Benefiting from this synergistic amplification, the platform achieved ultra-low limits of detection (LOD) of 153 fM (fluorescence), 516 fM (colorimetric), and 6.17 pM (smartphone) for the dual targets, coupled with exceptional specificity capable of discriminating single-base mismatches. The platform was successfully applied to detect miR-21 and miR-122 in mouse serum from a concanavalin A (ConA)‑induced AIH model, showing significantly elevated signals in model mice that were notably reduced after prednisone treatment, consistent with liver function indices and inflammatory cytokine profiles. This sensing strategy features simple operation, low cost, and no need for sophisticated instruments, providing a reliable point-of-care tool for AIH progression monitoring and therapeutic efficacy evaluation. It also offers a new methodological reference for multiplex biomarker detection in autoimmune diseases, showing promising potential for clinical translation. Download: Download high-res image (160KB) Download: Download full-size image This work presents a trimodal biosensor integrating D -CHA with CRISPR/Cas12a for sensitive detection of autoimmune hepatitis biomarkers miR-21 and miR-122. Target-triggered D -CHA activates Cas12a to cleave a G-rich reporter probe, enabling fluorescence, colorimetric, and smartphone-based RGB detection. The platform achieves fM-level sensitivity, single-base mismatch discrimination, and successful application in a mouse AIH model, offering a simple, low-cost POC tool for disease monitoring.

DNA logic-gated electrochemiluminescence biosensor for sensitive detection of dual-miRNA in bladder cancer

Jingjing Zhang,Yingxin Xie,Xiaoxu Wang,Xiaoman Ju,Lixing Weng,Lianhui Wang,Jie Chao,Lin Hao

Early diagnosis of bladder cancer requires highly sensitive and specific biosensors for detecting trace biomarkers in complex biological samples. However, single- biomarker detection is often prone to false-positive results due to biological variability and nonspecific fluctuations. Herein, we present a dual-target electrochemiluminescence (ECL) biosensor that integrates a DNA “AND” logic gate with toehold-mediated strand displacement (TMSD) circuits for parallel detection of miR-183–5p and miR-96–5p in human urine. A high-surface-area UIO-66@AuNPs composite served as the electrode substrate, increasing the loading of DNA logic gates and thus the Ru(bpy) 3 2 + - labeled signal probes, which significantly enhanced ECL response. The constructed DNA “AND” logic gate, consisting of a Ru(bpy) 3 2+ -labeled signal strand, a ferrocene-quenched strand, and a hairpin probe, restored the ECL signal only when both target miRNAs triggered parallel TMSD reactions, thereby achieving “AND” logic operation. This dual-target strategy effectively suppresses false-positive signals arising from single-biomarker fluctuations, thereby improving diagnostic accuracy compared with conventional single-target assays. Benefiting from TMSD-mediated circuits, the biosensor achieved detection limits as low as 0.45 fM for miR-183–5p and 0.85 fM for miR-96–5p over a broad linear range of 1 fM to100 pM. Urine analysis revealed significantly elevated levels of these miRNAs in bladder cancer patients. Furthermore, the quantified miRNA concentrations were incorporated into a machine learning-based classification model, enabling accurate discrimination between healthy individuals and bladder cancer patients. This integration of logic-gated biosensing with data-driven analysis not only improves diagnostic reliability but also demonstrates a promising strategy for intelligent biosensing in biological analysis and clinical auxiliary diagnosis.

Electrochemical sensor for miRNA-34a detection: Synergistic signal amplification for robust and sensitive analysis

Yanjie Zheng,Yibin Cai,Siyi Wang,Chunyan Liu,Kun Yang,Wanrong Chen,Hui Zhang,Jie Tan,Yaobin Chen,Shihao Zhuang,Wei Chen,Feng Feng,Chao Teng

This study developed an electrochemical biosensor for the detection of miRNA-34a by integrating a synergistic signal amplification strategy that combined the electrocatalytic properties of reduced graphene oxide (rGO) with the enzymatic activity of double-stranded specific nuclease (DSN), while employing magnetic separation technology to ensure robust interference resistance. The biosensor was constructed by electrodepositing rGO onto a glassy carbon electrode via cyclic voltammetry (CV), utilizing its electrocatalytic activity to enhance the electrochemical response. Simultaneously, the presence of target miRNA-34a initiated a DSN-mediated enzymatic cyclic reaction, resulting in secondary signal amplification. The incorporation of magnetic nanoparticle separation technology effectively reduced nonspecific absorption and background interference, enabling accurate determination of the target miRNA. The biosensor demonstrated exceptional detection performance, achieving quantitative detection of miRNA-34a across a broad linear range (50.0 fM to 1.0 nM) with a detection limit of 16.6 fM. In spike-recovery assays using human plasma samples, the sensor reliably measured miRNA-34a at varying concentrations, confirming its robust anti-interference capability in complex biological matrices. These findings suggest that the proposed biosensor has significant potential for clinical diagnostics and real-sample analysis applications.

Electrospun miRNA-21@doxycycline core-sheath nanofibers for a synergistic treatment of diabetic wounds

Minghui Xu,Xiaoru Yao,Yichen Wang,Haihan Liu,Wenjia Yang,Pingping Zhang

In this paper, core-sheath nanofiber membrane (miRNA-21@DCH) co-loaded with microRNA-21 and doxycycline (DCH) was prepared by coaxial electrospinning technology for the treatment of diabetic wounds. DCH could inhibit the growth of Gram-negative and Gram-positive bacteria that easily cause infection at the wound site, thus promoting the epidermal layer thickening and dermal connective tissue hyperplasia. The cumulative release rate of DCH was 89 % after 72 h. In vitro antibacterial experiments at 24 h showed that the bacteriostatic zone diameter of miRNA-21@DCH nanofiber membrane was about 3.82 cm for Escherichia coli and 4.54 cm for Staphylococcus aureus , indicating that miRNA-21@DCH nanofiber membrane had good antibacterial effect. The microRNA-21 could promote angiogenesis and collagen deposition at the wound site, providing a favorable growth environment for tissue cell proliferation. The synergistic effect of DCH and microRNA-21 confers miRNA-21@DCH core-sheath nanofiber membrane the advantage of rapid healing of diabetic wounds. After 16 days treatment with miRNA-21@DCH nanofiber membrane, the wound basically healed and the wound surface was smooth. In addition, in vitro cytotoxicity assays showed that miRNA-21@DCH nanofiber membrane had good biocompatibility. The prepared miRNA-21@DCH nanofiber membrane provides a new and effective strategy for the synergistic treatment of diabetic wounds.

Efficient miRNA Inhibitor Delivery with Graphene Oxide-Polyethylenimine to Inhibit Oral Squamous Cell Carcinoma

Lingling Ou,Ting Sun,Minyi Liu,Ye Zhang,Zhiying Zhou,Xiaozhen Zhan,Lihong Lu,Qingtong Zhao,Renfa Lai,Longquan Shao

Background MicroRNAs (miRNAs) are widely believed to be promising targets for oral squamous cell carcinoma (OSCC) gene therapy. miR-214 has been identified as a promoter of OSCC aggression and metastasis. Methods Graphene oxide-polyethylenimine (GO-PEI) complexes were prepared and loaded with a miRNA inhibitor at different N/P ratios. The transfection efficiency of GO-PEI-inhibitor was tested in Cal27 and SCC9 cells. Moreover, the tumor inhibition ability of GO-PEI-inhibitor was measured in an OSCC xenograft mouse model by intratumoral injection. Results Here, we show that a GO-PEI complex efficiently delivers a miR-214 inhibitor into OSCC cells and controls the intracellular release of the miR-214 inhibitor. These results indicate that the GO-PEI-miR-214 inhibitor complex efficiently inhibited cellular miR-214, resulting in a decrease in OSCC cell invasion and migration and an increase in cell apoptosis by targeting PTEN and p53. In the xenograft mouse model, the GO-PEI-miR-214 inhibitor complex significantly prevented tumor volume growth. Conclusion This study indicates that functionalized GO-PEI with low toxicity has promising potential for miRNA delivery for the treatment of OSCC.