SENSORS AND ACTUATORS B-CHEMICAL
·
2021
·
IF 7.46
Yunpeng Xu,Yanzi Zhang,Xiaolu Sui,Aisha Zhang,Xinguang Liu,Zhenyu Lin,Jihong Chen
MicroRNA-21(miRNA-21) can act as a novel biomarker in early stage of acute kidney injury (AKI). The development of simple but sensitive miRNA-21 detection technology is helpful for prevention and treatment of AKI. In this work, based on the cascaded signal amplifications of the target-induced hybridization chain reaction (HCR) and magnetic assisted enrichment, a highly sensitive homogeneous electrochemiluminescence (ECL) biosensor for miRNA-21 detection had been developed. Firstly, capture probe H1 (CPH1) had been modified on magnetic beads (MBs) which could capture target (miRNA-21) and trigger HCR amplification to produce long strand dsDNA in the homogeneous solution. Ru(phen) 3 2+ could be embedded into the grooves of dsDNA and played the role of luminous tag, which could be separated by magnetic easily to realize the enrichment. Then the enriched products had been dissolved in the buffer solution and the ECL signal could be detected easily. The ECL intensity had a linear correlation with the logarithm of miRNA-21 concentration in the range of 1.0 fM-1.0 nM, the detection limit was 0.14 fM. The method developed in this work showed well sensitivity, specificity, which had been applied to detect miRNA-21 in urine of AKI patients with satisfied results.
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高影响因子
MICROCHIMICA ACTA
·
2019
·Vol. 186
·期 11
·1-9
·
IF 5.479
Liu Chao,Zhao Li,Liang Dongxia,Zhang Xiaoru,Song Weiling
A photocathode is described for the determination of microRNA-21 by using CuInS 2 as an active photocathode material. Exonuclease III assisted target recycling amplification was employed to enhance the detection sensitivity. The TATA-binding protein (TBP) was applied to enhance steric hindrance which decreases the photoelectrochemical intensity. This strategy is designed by combining the anti-interference photocathode material, enzyme assisted target recycling amplification and TBP induced signal off, showing remarkable amplification efficiency. Under the optimized conditions, the detection limit for microRNA-21 is as low as 0.47 fM, and a linear range was got from 1.0 × 10 −15 M to 1.0 × 10 −6 M.
Schematic representation of sensitive photoelectrochemical detection of microRNA-21.CuInS 2 is used as an active photocathode material. Combined Exonuclease III assisted target recycling amplification and TATA-binding protein decreased of photoelectrochemical intensity, the detection limit was 0.47 fM with good selectivity. (miR-21: microRNA-21; CS: chitosan).
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高影响因子
CLINICAL AND EXPERIMENTAL PHARMACOLOGY AND PHYSIOLOGY
·
2018
·Vol. 46
·期 4
·389-397
·
IF 2.092
Hao Yuan,Hongyan Zhao,Jiafeng Wang,Hong Zhang,Lihua Hong,He Li,Hongze Che,Zhimin Zhang
Summary Pulpitis suppressed the level of let-7c-5p that promotes osteogenesis and bone formation by repressing HMGA2. In the current study, the function of let-7c-5p in the inflammation and osteogenesis in dental pulp stem cells (DPSCs) was explored. The level of let-7c-5p in DPSCs was up-regulated, and the cells were subjected to lipopolysaccharide (LPS) to induce inflammation. The effect of let-7c-5p on cell proliferation potential, osteogenic differentiation potential, and activity of HMGA2/PI3K/Akt pathway was detected. The administration of LPS suppressed the cell proliferation of DPSCs and suppressed calcium deposition, activity of alkaline phosphatase (ALP), and levels of OCN, OPN, OSX, MSX2, and RUNX2 in inflamed DPSCs. The impaired osteogenic differentiation of inflamed DPSCs was associated with the increased levels of HMGA2, p-PI3K, and p-Akt. In let-7c-5p-overexpressed inflamed DPSCs, the proliferation and osteogenic differentiation potential of DPSCs were restored, and the activation of HMGA2/PI3K/Akt signalling was inhibited. In rat pulpitis models, the injection of let-7c-5p agomir restored the osteogenic differentiation potential of dental pulp cells and inhibited HMGA2/PI3K/Akt signalling. The findings demonstrated the anti-inflammation and pro-osteogenesis effect of let-7c-5p during the attack of pulpitis, which depended on the inhibition of HMGA2/PI3K/Akt signalling.
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高影响因子
JOURNAL OF CELLULAR PHYSIOLOGY
·
2018
·Vol. 234
·期 3
·2537-2551
·
IF 3.923
Hai-Ming Xu,Feng-Hua Sui,Mei-Hua Sun,Gong-Liang Guo
Recent studies have shown that circulating microRNAs (miRNA) play a critical role in diagnosing acute coronary syndrome (ACS). This study aims to investigate the effect of miR-224 on atherosclerotic plaques forming and vascular remodeling in ACS and its relationship with TGF-β/Smad pathway. Myocardial infarction (MI) rat model was established and lentivirus vector of miR-224 inhibitor was prepared for investigating the effect of downregulated miR-224 on the contents of nitric oxide (NO) and endothelin-1 (ET-1), blood lipid levels and inflammatory factor levels in serum as well as the TGF-β/Smad pathway. The rats suffering from MI had decreased survival rates and exhibited reduced levels of NO, high-density lipoprotein cholesterol, and lumen diameter, and Smad7 messenger RNA (mRNA) and protein expression; while had significantly increased ratio of heart weight or body weight, levels of ET-1, inflammatory factors, blood lipid indexes, vascular remodeling indexes, collagen volume fraction, vulnerable atherosclerotic plaque area, VCAM-1 and MMP-2 protein expression, TGF-β, Smad2, Smad3, and Smad4 mRNA and protein expression. After inhibiting the TGF-β/Smad pathway, the rats suffering from MI showed notably opposite trend. In conclusion, downregulation of miR-224 expression promotes the formation of vulnerable atherosclerotic plaques and vascular remodeling in ACS through activation of the TGF-β/Smad pathway. Therefore, this study provides a new therapeutic target for ACS.
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BIOSENSORS & BIOELECTRONICS
·
2018
·
IF 8.173
Liang Tian,Jinxu Qi,Xiangyu Ma,Xuejiao Wang,Chen Yao,Wei Song,Yihong Wang
Herein, we report a facile enzyme-free microRNA (miRNA) target-triggered strand displacement reaction (SDR) amplification strategy with ferrocene (Fc) as a signal molecule to fabricate a two-dimensional electroactive molybdenum carbide (Mo 2 C)-based biosensor. In the presence of miRNA-21, SDR was initiated and many hairpin DNA1 (HDNA1) and hairpin DNA2 (HDNA2) duplexes, which could be captured by probe DNA leading the Fc-modified HDNA2 close to the electrode surface , were produced continuously. MiRNA-21 could be detected by monitoring the redox signal of Fc. The prepared N -carboxymethyl chitosan/Mo 2 C nanocomposite featured excellent conductivity , great dispersion, and multiple functional groups (amine groups). When the nanocomposite was introduced to a miRNA biosensor electrode interface to ensure its strong connection to the DNA probe, the developed miRNA-21 biosensor demonstrated a reliable linear range of 1.0 fM to 1.0 nM with a detection limit of 0.34 fM and showed good selectivity , reproducibility, and stability. The biosensor was employed to detect miRNA-21 in human serum samples, and it showed great potential in the early clinical diagnosis of various genetic diseases.
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MICROCHIMICA ACTA
·
2018
·Vol. 185
·期 4
·1-8
·
IF 5.705
Yu Xinsheng,Hu Lianzhe,Zhang Feng,Wang Min,Xia Zhining,Wei Weili
A dual-channel ratiometric nanoprobe is described for detection and imaging of microRNA. It was prepared from MoS 2 quantum dots (QDs; with blue emission and excitation/emission peaks at 310/398 nm) which acts as both the gene carrier and as a donor in fluorescence resonance energy transfer (FRET). Molecular beacons containing loops for molecular recognition of microRNA and labeled with carboxyfluorescein (FAM) were covalently attached to the MoS 2 QDs and serve as the FRET acceptor. In the absence of microRNA, the nanoprobe exhibits low FRET efficiency due to the close distance between the FAM tag and the QDs. Hybridization with microRNA enlarges the distance between QD and beacon. This results in an enhancement of the FRET efficiency of the nanoprobe. The ratio of green and blue fluorescence (I 520 /I 398 ) increases linearly in the 5 to 150 nM microRNA concentration range in both aqueous solution and diluted artificial cerebrospinal fluid. The limit of detection (LOD) is as low as 0.38 nM and 0.52 nM, respectively. Other features of this nanoprobe include (a) excellent resistance to nuclease-induced false positive signals and (b) the option to use it for distinguishing different cell lines by in-situ imaging of intracellular microRNAs.
Schematic of a dual-channel photoluminescence nanoprobe for the determination of microRNA-21 (miR-21) by monitoring the microRNA-triggered enhancement of the fluorescence resonance energy transfer (FRET) efficiency between MoS 2 QDs and carboxyfluorescein-labeled molecular beacons.
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高影响因子
BIOSENSORS & BIOELECTRONICS
·
2016
·
IF 7.476
Hong-Lei Shuai,Ke-Jing Huang,Ling-Li Xing,Ying-Xu Chen
An ultrasensitive electrochemical biosensor for microRNA (miRNA) is developed based on tungsten oxide-graphene composites coupling with catalyzed hairpin assembly target recycling and enzyme signal amplification . WO 3 -Gr is prepared by a simple hydrothermal method and then coupled with gold nanoparticles to act as a sensing platform. The thiol-terminated capture probe H1 is immobilized on electrode through Au–S interaction. In the presence of target miRNA, H1 opens its hairpin structure by hybridization with target miRNA. This hybridization can be displaced from the structure by another stable biotinylated hairpin DNA (H2), and target miRNA is released back to the sample solution for next cycle. Thus, a large amount of H1-H2 duplex is produced after the cyclic process. At this point, a lot of signal indicators streptavidin-conjugated alkaline phosphatase (SA-ALP) are immobilized on the electrode by the specific binding of avidin-biotin. Then, thousands of ascorbic acid , which is the enzymatic product of ALP, induces the electrochemical-chemical-chemical redox cycling to produce a strongly electrochemical response in the presence of ferrocene methanol and tris (2-carboxyethyl) phosphine. Under the optimal experimental conditions, the established biosensor can detect target miRNA down to 0.05 fM (S/N=3) with a linear range from 0.1 fM to 100 pM, and discriminate target miRNA from mismatched miRNA with a high selectivity .
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高影响因子
BIOSENSORS & BIOELECTRONICS
·
2012
·
IF 5.602
Huanshun Yin,Yunlei Zhou,Haixia Zhang,Xiaomeng Meng,Shiyun Ai
MicroRNAs (miRNAs), a kind of small, endogenous, noncoding RNAs (∼22 nucleotides), might play a crucial role in early cancer diagnose due to its abnormal expression in many solid tumors. As a result, label-free and PCR-amplification-free assay for miRNAs is of great significance. In this work, a highly sensitive biosensor for sequence specific miRNA-21 detection without miRNA-21 labeling and enrichment was constructed based on the substrate electrode of dendritic gold nanostructure (DenAu) and graphene nanosheets modified glassy carbon electrode. Sulfydryl functionalized locked nucleic acid (LNA) integrated hairpin molecule beacon (MB) probe was used as miRNA-21 capture probe. After hybridized with miRNA-21 and reported DNA loading in gold nanoparticles (AuNPs) and biotin multi-functionalized bio bar codes, streptavidin–HRP was brought to the electrode through the specific interaction with biotin to catalyze the chemical oxidation of hydroquinone by H 2 O 2 to form benzoquinone. The electrochemical reduction signal of benzoquinone was utilized to monitor the miRNA-21 hybridization event. The effect of experimental variables on the amperometric response was investigated and optimized. Based on the specific confirmation of probe and signal amplification, the biosensor showed excellent selectivity and high sensitivity with low detection limit of 0.06 pM. Successful attempts are made in miRNA-21 expression analysis of human hepatocarcinoma BEL-7402 cells and normal human hepatic L02 cells.
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高影响因子
BIOSENSORS & BIOELECTRONICS
·
2018
·
IF 8.173
Liang Tian,Jinxu Qi,Olayinka Oderinde,Chen Yao,Wei Song,Yihong Wang
Enzyme mimics have been developed for bioassay of nucleic acids , with some of them involving complicated labeling. Herein, we report a label-free bioassay for ultrasensitive electronic determination of microRNA at an ultralow concentration based on target-triggered long-range self-assembly DNA-based hybridization chain reaction (HCR) protocol coupled with bienzyme mimics synergistic catalysis strategy. In this work, a planar intercalation molecule, copper (II) complex, is applied for the first time as a small molecule enzyme mimic as well as intercalation molecule in microRNA biosensor for signal amplification . Fe 3 O 4 nanozyme were used as a separate and enriched target under magnetic field, and also in combination with HCR protocol detected in 3,3’,5,5’-tetramethylbenzidine+hydrogen peroxide (TMB+H 2 O 2 ) system to improve the sensitivity of the biosensor. Under optimal conditions, these strategies present good electrochemical behaviors for the detection of microRNA with a wide range from 100 aM to 100 nM and at relatively low detection limit of 33 aM This remarkable sensitivity can make this proposed approach a promising scheme for development of next-generation microRNA sensors without the need of enzyme labeling or fluorophore labeling.
研究领域:
生物化学
·
分子生物学
·
分析化学
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SENSORS AND ACTUATORS B-CHEMICAL
·
2022
·
IF 9.221
Li Shuofeng,Wang Fangfang,Chun Wang,Zhi Wang,Qiuhua Wu
As vital biomarkers for diagnosis of diseases, the anomalously expressed microRNAs (miRNAs) are closely linked to the occurrence and development of many diseases. Quantitative analysis of miRNAs has remarkable significance to clinical diagnosis and therapy of different cancers. In this study, the clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated (Cas) 13a (CRISPR/Cas13a) system was first exploited to directly and specifically recognize the miRNA at the guidance of CRISPR RNA (crRNA), which can activate the nonspecific trans-ribonuclease activity of Cas13a. Then, combining the CRISPR-Cas13a system with terminal deoxynucleotidyl transferase (TdT)-triggered template-free DNA extension to form the fluorescent copper nanoparticles (CuNCs), a simple but sensitive method (named as Cas13a-TFCN) was developed for highly specific detection of miRNA. Based on the CRISPR/Cas13a system coupled with TdT-triggered simple but efficient signal amplification strategy, the developed Cas13a-TFCN method showed high sensitivity for miRNA detection with the low detection limit of 0.7 fM. Furthermore, the proposed Cas13a-TFCN approach was successfully applied for accurate quantification of miRNA in complex biological samples, indicating its practical application ability. Moreover, featuring high sensitivity and simplicity without complicated probe design and fluorophore labeling, the proposed Cas13a-TFCN has a promising applications prospect in clinical diagnosis.
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ANALYTICAL CHEMISTRY
·
2020
·
IF 6.785
Jia Jun Liu,Hui Hong Yan,Qiang Zhang,Peng Fei Gao,Chun Mei Li,Gao Lin Liang,Cheng Zhi Huang,Jian Wang
As an important biomarker for early diagnosis of cancers, sensitive detection and high-resolution imaging of microRNA-21 in cancer cells have become important and challengeable. In this work, highly sensitive detection and spatial imaging of intracellular microRNA-21 were realized by the reduced signal background through vertical polarization excitation with a polarizer. The lateral local surface plasmon resonance property of gold nanorods (AuNRs) displayed a pronounced green color with low scattering intensity, which was adjusted to red color with strong scattering intensity when the core–satellite gold nanoparticle (AuNP) assembly was constructed on the side of AuNRs through a catalyzed hairpin assembly (CHA) circuit in the presence of microRNA-21. This unique approach allows for effectively reducing the strong background signal to improve the sensitivity of detection. Additionally, the proposed strategy can not only realize the sensitive detection of microRNA-21 with the limit of detection as low as 2 pM (3σ) but also achieve the high spatial imaging of cancer cells, which provided a specific strategy for the construction and imaging of intracellular imaging probes. It is believed that the simple and sensitive approach on the basis of lateral local surface plasmon resonance property of anisotropic AuNRs with excellent sensitivity combined with high spatial imaging holds promising potentials to visualize intracellular microRNAs with low abundance.
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高影响因子
ANALYTICA CHIMICA ACTA
·
2020
·
IF 5.977
Xiaotong Wei,Dewei Liu,Min Zhao,Tiantian Yang,Yunpeng Fan,Wenqin Chen,Ping Liu,Jianbo Li,Shijia Ding
MicroRNAs (miRNAs), considered as therapeutic targets and biomarkers, play important roles in biological processes . Herein, an enzyme-free surface plasmon resonance imaging (SPRi) biosensing method has been developed for miRNA detection based on catalytic hairpin assembly and spherical nucleic acid . The hairpin H1 tethered on the surface of the sensor chip is unfolded by miRNA, and then the hybridized miRNA is released through the displacement of the hairpin H2 for the successive hybridization and assembly process. The emerging DNA fragments on the sensor chip surface after hairpins assembly are further used to hybridize with spherical nucleic acid, inducing a remarkably amplified SPR signal. This biosensing method is highly sensitive to miRNA with a detection limit of 53.7 fM and a linear range of 4 orders of magnitude. Moreover, the biosensor demonstrates good specificity and has the ability to distinguish members of homologous miRNA family even with single base differences. Thus, the SPRi biosensing method may hold a great promise for further application in early clinical diagnosis.
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