ANALYTICAL CHEMISTRY
·
2014
·
IF 5.825
Mo Wang,Zhengliang Fu,Bingchen Li,Yunlei Zhou,Huanshun Yin,Shiyun Ai
Taking advantage of the special exodeoxyribonuclease activity of T7 exonuclease, a simple, sensitive, selective, and label-free microRNA biosensor based on the cyclic enzymatic amplification method (CEAM) has been proposed. First, thiol functionalized DNA probes were assembled onto a gold nanoparticles modified gold electrode surface through a Au–S bond, followed by hybridizing with target miRNA. Subsequently, DNA in RNA/DNA duplexes was digested by T7 exonuclease, which can release the microRNA molecules from the electrode surface and return into the buffer solution. Meanwhile, the released microRNA can further hybridize with the unhybridized DNA probes on the modified electrode surface. On the basis of it, an isothermal amplification cycle is realized. The T7 exonuclease-assisted CEAM achieved a low detection limit of 0.17 fM. Moreover, this assay presents excellent specificity with discriminating only a single-base mismatched microRNA sequence. Furthermore, this work can also be applied to detect avian leukemia based on the decreased expression level of microRNA-21.
研究领域:
生物化学
·
分子生物学
·
分析化学
·
癌症研究
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产品关联
DOI
PubMed
高影响因子
ENVIRONMENTAL POLLUTION
·
2021
·
IF 8.071
Weikai Ding,Yingying Shangguan,Yuqing Zhu,Yousef Sultan,Yiyi Feng,Bangjun Zhang,Yang Liu,Junguo Ma,Xiaoyu Li
Microcystin-LR (MC-LR) and glyphosate (GLY) have been classified as a Group 2B and Group 2A carcinogens for humans, respectively, and frequently found in aquatic ecosystems . However, data on the potential hazard of MC-LR and GLY exposure to the fish gut are relatively scarce. In the current study, a subacute toxicity test of zebrafish exposed to MC-LR (35 μg L −1 ) and GLY (3.5 mg L −1 ), either alone or in combination was performed for 21 d. The results showed that MC-LR or/and GLY treatment reduced the mRNA levels of tight junction genes ( claudin-5 , occludin , and zonula occludens-1 ) and altered the levels of diamine oxidase and D-lactic, indicating increased intestinal permeability in zebrafish. Furthermore, MC-LR and/or GLY treatment remarkably increased the levels of intestinal IL-1β and IL-8 but decreased the levels of IL-10 and TGF-β, indicating that MC-LR and/or GLY exposure induced an inflammatory response in the fish gut. MC-LR and/or GLY exposure also activated superoxide dismutase and catalase , generally upregulated the levels of p53 , bax , bcl-2 , caspase-3 , and caspase-9 , downregulated the levels of caspase-8 and caused notable histological injury in the fish gut. Moreover, MC-LR and/or GLY exposure also significantly altered the microbial community in the zebrafish gut and the expression of miRNAs (miR-146a, miR-155, miR-16, miR-21, and miR-223). Chronic exposure to MC-LR and/or GLY can induce intestinal damage in zebrafish, and this study is the first to demonstrate an altered gut microbiome and miRNAs in the zebrafish gut after MC-LR and GLY exposure.
研究领域:
毒理学
·
环境科学
·
分子生物学
·
鱼类生物学
·
微生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2017
·
IF 7.78
Shuzhen Yue,Tingting Zhao,Sai Bi,Zhipeng Zhang
High throughput analysis of miRNAs is of great significance to clinical and biomedical applications . In this work, we have developed a highly sensitive and selective chemiluminescence imaging array (CLIA) for simultaneous detection of three miRNAs with high throughput, easy operation and low cost. In this assay, three kinds of hairpins that can specially recognize respective miRNAs (miR-155, miR-let-7a and miR-141) are parallel modified on the magnetic beads (MBs). The employment of MBs enables quick sorting of multiplex targets in high yield and purity with high throughput. Upon introduction of the target miRNAs, they hybridize to the corresponding hairpins and initiate the DNA machines with the assistance of Klenow fragment exo- DNA polymerase and Nb.BbvCI NEase , achieving exponential amplification of targets. After that, the toehold-mediated strand displacement (TMSD) reactions are performed through sequential addition of displacement probes under magnetic separation, resulting in the release of HRP-tagged DNA hybrids in solution for CL imaging and recovery of MBs for repetitive use. This CLIA method demonstrates ultrahigh sensitivity with detection limits down to fM level, wide linear range over 5 orders of magnitude, excellent selectivity to distinguish one-base mismatched target miRNA, and good performance in real sample analysis. Together with the advantages of high throughput, easy operation, acceptable accuracy, and good recyclability and reproducibility, the proposed multiplex CLIA method holds great potential in practical applications, such as profiling the pattern of miRNA expression, disease screening, biomedical research, and so on.
研究领域:
生物医学工程
·
分子诊断学
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DOI
PubMed
高影响因子
ANALYST
·
2021
·Vol. 146
·期 10
·3242-3250
·
IF 4.616
Pan Fu,Mengjia Xu,Shu Xing,Yang Zhao,Chao Zhao
The practical use of a point-of-care (POC) device is of particular interest in performing liquid biopsies related to cancer. Herein, taking advantage of the practical convenience of a commercially available personal glucose meter (PGM), we report a convenient, low-cost and sensitive detection strategy for circulating microRNA-155 (miRNA155) in human serum. First, miRNA155 in serum triggers the catalyzed hairpin assembly (CHA) reaction, and then the CHA product is specifically captured by the peptide nucleic acid (PNA) probes attached to the surface of a 96-well plate, which in turn triggers the hybridization chain reaction (HCR), resulting in the local enrichment of invertase. Next, introduction of a substrate (sucrose) for the invertase results in the generation of glucose, which can be detected by a PGM. In this sensor, neutrally charged PNA (12 nt) is more likely to hybridize with the CHA products than with the negatively charged DNA in kinetics, which improves the detection sensitivity and specificity. Due to the synergistic isothermal amplification reaction between CHA and HCR, the sensor is able to achieve a broad dynamic range (from 1 fM to 10 nM) with a detection limit down to 0.36 fM (3 orders of magnitude lower than that without HCR) and is capable of distinguishing single-base mismatched sequences. Thus the convenient, sensitive, robust and low-cost PGM sensor makes on-site nucleic acids detection possible, suggesting its great application prospect as a promising POC device in cancer diagnostics.
研究领域:
医学诊断
·
生物技术
·
纳米技术
·
分子生物学
·
癌症研究
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DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2022
·
IF 8.008
Kefeng Wu,Qiang Li,Chi Yao,Dayong Yang,Dingbin Liu
We report a Trojan horse strategy to efficiently deliver the spherical nucleic acid probes (namely, nanoflares) into the cytoplasm for microRNA (miRNA) imaging with high fidelity, breaking through the cytoplasmic transport dilemma of RNA probes in living cells. The nanoflare is encapsulated into a “Trojan horse” consisting of zwitterionic choline phosphates (CPs) and acid-degradable crosslinkers; the former effectively promotes cell uptake and the latter triggers instantaneous liberation of the nanoflare probes from the lysosome to the cytoplasm. The exposed nanoflares in the cytoplasm can be lightened up by the target miRNAs specifically. Compared with the conventional nanoflares as well as the improved ones in previous reports, the “Trojan horse” nanoflares avoid nuclease degradation and thiol displacement during the delivery process, providing unprecedentedly high accuracy for intracellular miRNA imaging.
研究领域:
分子生物学
·
纳米技术
·
生物医学工程
·
细胞生物学
✓ 已验证
DOI
PubMed
高影响因子
Gut Microbes
·
2022
·
IF 9.434
Xihong Zhou,Yonghui Liu,Xia Xiong,Jingqing Chen,Wenjie Tang,Liuqin He,Zhigang Zhang,Yulong Yin,Fengna Li
Diarrheal disease is a common health problem with complex causality. Although diarrhea is accompanied by disturbances in microbial diversity, how gut microbes are involved in the occurrence of diarrhea remains largely unknown. Here, using a pig model of post-weaning stress-induced diarrhea, we aim to elucidate and enrich the mechanistic basis of diarrhea. We found significant alterations in fecal microbiome, their metabolites, and microRNAs levels in piglets with diarrhea. Specifically, loss of ssc-miRNA-425-5p and ssc-miRNA-423-3p, which inhibit the gene expression of fumarate reductase (frd) in Prevotella genus, caused succinate accumulation in piglets, which resulted in diarrhea. Single-cell RNA sequencing indicated impaired epithelial function and increased immune response in the colon of piglet with diarrhea. Notably, the accumulated succinate increased colonic fluid secretion by regulating transepithelial Cl-secretion in the epithelial cells. Meanwhile, succinate promoted colonic inflammatory responses by activating MyD88-dependent TLR4 signaling in the macrophages. Overall, our findings expand the mechanistic basis of diarrhea and suggest that colonic accumulation of microbiota-produced succinate caused by loss of miRNAs leads to diarrhea in weanling piglets.
研究领域:
兽医学
·
微生物学
·
分子生物学
·
胃肠病学
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DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2026
·Vol. 98
·期 31
·23153-23163
·
IF 7.3
Peng Ruixiao,Xiao Weirong,Xu Xiaodong,Liu Sinong,Zhang Yuqian,Tian Wenhai,Yang Chaoyong,Song Yanling,Cui Liang
Extracellular vesicles (EVs) have
emerged as a prominent
and rapidly
evolving class of biomarkers, garnering an increasing amount of attention
in recent years. The analysis of diverse EV-derived biomolecules,
including proteins, mRNA, and miRNA, has driven the development of
numerous innovative analytical technologies for EV detection. Nevertheless,
despite significant advancements, most current EV analysis methods
still require substantial improvements to address critical limitations,
such as operational complexity, insufficient sensitivity, and the
persistent issue of false-positive signals. In this study, we present
a targeted fusion and nonenzymatic amplification strategy for ultrasensitive
detection of tumor-derived EV miRNAs. The proposed approach integrates
an aptamer-anchored liposome system that facilitates specific recognition
and fusion with tumor EVs, coupled with a spatial confinement nonenzymatic
signal amplification system to significantly enhance detection sensitivity.
In conjunction with flow cytometry, the platform demonstrates a detection
limit of 4 ng/mL for EV- associated proteins (0.2 fM for EV miR-21)
and effectively discriminates cancer patients from healthy controls.
This innovative strategy shows substantial promise for diverse biomedical
applications, potentially revolutionizing early cancer diagnosis,
metastasis monitoring, and prognostic evaluation while also offering
opportunities for the detection and management of various other diseases.
✓ 已验证
产品关联
DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2026
·
IF 8.7
Meijiao Duan,Qiyue Sun,Jiaqi Duan,Zhuo Chen,Chunyan Yang,Cui Zhai,Meng Cao,Huan Liu,Mengqian Wei,Yunong Chen,Lei Lei,Xingchun Gou,Liujiao Bian,Jing Luan
This study presents a groundbreaking investigation into the molecular interaction between microRNAs (miRNAs) from the traditional Chinese medicine Astragali Radix and the anti-angiogenic protein Kringle 5 (K5). We successfully screened and identified 10 highly enriched miRNAs that exhibit a significant binding affinity to K5. Through a combination of bioinformatics and experimental techniques, including molecular docking and affinity chromatography, we found that specific miRNAs, particularly miR1, act as natural aptamers, binding to K5 with high selectivity and an association constant ( K a ) of approximately 5.80 × 10 4 L/mol. Our findings reveal that the conserved stem-ring structure of these miRNAs is crucial for their recognition and binding to K5, mediated by electrostatic, hydrophobic, and hydrogen bond interactions. Functional assays in vitro demonstrate that miR1 effectively neutralizes K5-mediated inhibition of endothelial cell proliferation and restores the HIF-1α/VEGF signaling pathway. This research provides a novel biochemical perspective suggesting that Astragalus -derived miRNAs may function as natural molecular decoys to modulate K5 activity, thereby supporting angiogenesis. These findings offer new insights into the bioactive landscape of traditional Chinese medicine and providing a strong foundation for the development of novel nucleic acid-based drugs. Download: Download high-res image (114KB) Download: Download full-size image
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DOI
PubMed
高影响因子
ACS Omega
·
2026
·
IF 5.2
YanYan Ma,Mingzhen Yang,Jiali Zheng,YiXin Zeng,Xuan Zhang,Juan Sun,Yushan Chen,Enqi Tang,Xiaolei Cai,Yaping Feng,Wenwang Li,Lei Guo,Zhe Huang,Xiuli Zhang,Cuimin Wu,Ying Sun
Early diagnosis of hepatocellular carcinoma is crucial for improving patient prognosis, and microRNA (miRNA) detection via liquid biopsy is important. However, the low abundance, high homology, and complex background interference of miRNAs in biological samples pose serious challenges to the sensitivity and specificity of their detction techniques. This study developed a catalytic hairpin assembly (CHA) signal amplification platform with dual activation of near-infrared light (NIR) and glutathione (GSH), combined with the low background fluorescence properties of upconversion nanoparticles (UCNPs) for the accurate and specific quantification of both miRNA-21 and miRNA-221. Under optimized conditions, the limits of detection for miRNA-21 and miRNA-221 were 0.266 and 0.113 nM, respectively. The recovery rates in the serum spiked experiments ranged from 98.0 to 107.1%, with a low relative standard deviation (<4.84%), suggesting that the method has good accuracy and reliability. The constructed NIR/GSH dual-activation CHA-sensing platform not only offers a novel approach for the determination of low-abundance miRNAs in complex biological samples but also establishes a feasible foundation for the development of advanced theranostic nanoplatforms for early screening and subtyping diagnosis of hepatocellular carcinoma.
✓ 已验证
DOI
PubMed
高影响因子
ACTA PAEDIATRICA
·
2021
·Vol. 110
·期 7
·2221-2229
·
IF 2.299
Jing Li,Meixue Chen,Jinfeng Wang,Lingling Lu,Xiang Li,Yuan Le
Aim This study aimed to profile the microRNA levels in Chinese Henoch-Schonlein purpura (HSP) children and to explore their association with inflammatory factors and T helper 17 (Th17)/regulatory T (Treg). Methods Forty-five HSP children and 27 healthy controls were enrolled in this study, and microRNA levels were profiled with a microRNA microarray. The levels of selected microRNAs were determined by quantitative real-time PCR, and the levels of serum IgA, interleukin-6, interleukin-10 and interleukin-17A were detected by enzyme-linked immunosorbent assay. Additionally, Th17 and Treg cells were analysed by flow cytometry. Results There were 9 up-regulated and 27 down-regulated microRNAs in the PBMCs of Chinese HSP children. Among them, miR-1-3p, miR-19b-1-5p and miR-29b-1-5p were up-regulated, while miR-483-5p and miR-1246 were down-regulated. Additionally, these selected microRNAs could differentiate HSP patients from healthy controls. Interestingly, miR-29b-1-5p was correlated with IgA, miR-19b-1-5p, miR-483-5p and miR-1246 were correlated with interleukin-6, while miR-1-3p and miR-1246 were correlated with Th17/Treg. Conclusion This study reveals that the altered microRNAs could differentiate HSP from the healthy, and were associated with inflammatory factors or Th17/Treg. It is indicated that alteration in these microRNAs may contribute to the HSP pathogenesis and may become therapeutic targets or diagnostic biomarkers for HSP.
研究领域:
儿科
·
免疫学
·
分子生物学
✓ 已验证
DOI
PubMed
高影响因子
JOURNAL OF APPLIED TOXICOLOGY
·
2020
·Vol. 41
·期 8
·1275-1285
·
IF 2.997
Shan Yi,Yan-ping Liu,Xun-ya Li,Xiao-yan Yuan,Ye Wang,Ying Cai,Yuan-di Lei,Lian Huang,Zhao-hui Zhang
Beryllium and its compounds are systemic toxicants that mainly accumulate in the lungs. As a regulator of gene expression, microRNAs (miRNAs) were involved in some lung diseases. This study aimed to analyze the levels of some inflammatory cytokine and the differential expressions of miRNAs in human bronchial epithelial cells (16HBE) induced by beryllium sulfate (BeSO 4 ) and to further explore the biological functions of differentially expressed miRNAs. The profile of miRNAs in 16HBE cells was detected using the high-throughput sequencing between the control groups ( n = 3) and the 150 μmol/L of BeSO 4 -treated groups ( n = 3). Bioinformatics analysis of differentially expressed miRNAs was performed, including the prediction of target genes, Gene Ontology (GO) analysis, and the Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis. Quantitative real-time polymerase chain reaction (qRT-PCR) was applied to verify some damage-related miRNAs. We found that BeSO 4 can increase the levels of some inflammatory cytokine such as interleukin-10 (IL-10), tumor necrosis factor-alpha (TNF-α), interferon-γ (IFN-γ), inducible nitric oxide synthase (iNOS), and cyclooxygenase-2 (COX-2). And BeSO 4 altered miRNAs expression of 16HBE cells and a total of 179 differentially expressed miRNAs were identified, including 88 upregulated miRNAs and 91 downregulated miRNAs. The target genes predicted by 28 dysregulated miRNAs were mainly involved in the transcription regulation, signal transduction, MAPK, and VEGF signaling pathway. The qRT-PCR verification results were consistent with the sequencing results. miRNA expression profiling in 16HBE cells exposed to BeSO 4 provides new insights into the toxicity mechanism of beryllium exposure.
研究领域:
毒理学
·
分子生物学
·
生物信息学
·
细胞生物学
✓ 已验证
DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2020
·
IF 6.785
Yuqiang Xiang,Huaduo Yan,Bingjie Zheng,Aroosha Faheem,Yonggang Hu
High-throughput analyses of multitarget markers can facilitate rapid and accurate clinical diagnosis. Suspension array assays, a flow cytometry-based analysis technology, are among some of the most promising multicomponent analysis methods for clinical diagnostics and research purposes. These assays are appropriate for examining low-volume, complex samples having trace amounts of analytes due to superior elimination of background. Physical shape is an important and promising code system, which uses a set of visually distinct patterns to identify different assay particles. Here, we presented a morphology recognizable suspension arrays based on the microorganisms with different morphologies. In this study, UiO-66-NH2 (UiO stands for University of Oslo) metal–organic frameworks (MOFs), was wrapped on the microorganism surface to form an innovative class of microorganism@UiO-66-NH2 composites for suspension array assays. The use of microorganisms endowed composites barcoding ability with their different morphology and size. Meanwhile, the UiO-66-NH2 provided a stable rigid shell, large specific surface area, and metal(IV) ions with multiple binding sites, which could simplify the protein immobilization procedure and enhance detection sensitivity. With this method, simultaneous detection of three colorectal cancer-related microRNA (miRNA), including miRNA-21, miRNA-17, and miRNA-182, could be easily achieved with femtomolar sensitivity by using a commercial flow cytometer. The synergy between microorganisms and MOFs make the composites a prospective barcoding candidate with excellent characteristics for multicomponent analysis, offering great potential for the development of high throughput and accurate diagnostics.
研究领域:
生物医学工程
·
分子生物学
·
癌症研究
·
分析化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）