BIOSENSORS & BIOELECTRONICS
·
2016
·
IF 7.476
Hong-Lei Shuai,Ke-Jing Huang,Ying-Xu Chen,Lin-Xia Fang,Meng-Pei Jia
An ultrasensitive electrochemical biosensor for detecting microRNAs is fabricated based on hollow molybdenum disulfide (MoS 2 ) microcubes. Duplex-specific nuclease , enzyme and electrochemical–chemical–chemical redox cycling are used for signal amplification . Hollow MoS 2 microcubes constructed by ultrathin nanosheets are synthesized by a facile template-assisted strategy and used as supporting substrate. For biosensor assembling, biotinylated ssDNA capture probes are first immobilized on Au nanoparticles (AuNPs)/MoS 2 modified electrode in order to combine with streptavidin-conjugated alkaline phosphatase (SA-ALP). When capture probes hybridize with miRNAs, duplex-specific nuclease cleaves the formative duplexes. At the moment, the biotin group strips from the electrode surface and SA-ALP is incapacitated to attach onto electrode. Then, ascorbic acids induce the electrochemical–chemical–chemical redox cycling to produce electrochemical response in the presence of ferrocene methanol and tris (2-carboxyethyl) phosphine. Under optimum conditions, the proposed biosensor shows a good linear relationship between the current variation and logarithm of the microRNAs concentration ranging from 0.1 fM to 0.1 pM with a detection limit of 0.086 fM (S/N=3). Furthermore, the biosensor is successfully applied to detect target miRNA-21 in human serum samples.
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高影响因子
BIOSENSORS & BIOELECTRONICS
·
2024
·
IF 10.7
Yuchen Song,Changqing Mao,Wenjiao Zhang,Dongmei Deng,Huinan Chen,Pei Sun,Meiyin Liu,Chang Feng,Liqiang Luo
The low abundance, heterogeneous expression, and temporal changes of miRNA in different cellular locations pose significant challenges for both the detection sensitivity of miRNA liquid biopsy and intracellular imaging. In this work, we report an intelligently assembled biosensor based on catalytic hairpin assembly (CHA) and aggregation-induced emission (AIE), named as catalytic hairpin aggregation-induced emission (CHAIE), for the ultrasensitive detection and intracellular imaging of miRNA-155. To achieve such goal, tetraphenylethylene-N 3 (TPE-N 3 ) is used as AIE luminogen (AIEgen), while graphene oxide is introduced to quench the fluorescence. When the target miRNA is present, CHA reaction is triggered, causing the AIEgen to self-assemble with the hairpin DNA . This will restrict the intramolecular rotation of the AIEgen and produce a strong AIE fluorescence. Interestingly, CHAIE does not require any enzyme or expensive thermal cycling equipment, and therefore provides a rapid detection. Under optimal conditions, the proposed biosensor can determine miRNA in the concentration range from 2 pM to 200 nM within 30 min, with the detection limit of 0.42 pM. The proposed CHAIE biosensor in this work offers a low background signal and high sensitivity, making it applicable for highly precise spatiotemporal imaging of target miRNA in living cells.
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高影响因子
Bioengineered
·
2021
·
IF 3.269
Jing Wang,Yan Liu,Hongbing Cai,Hong Jiang,Wei Li,Yuying Shi
Cervical cancer (CC) is one of the most common malignancies among women. It has been demonstrated that long coding RNAs (lncRNAs) play a crucial role in CC. The purpose of this study was to investigate the role of the colon cancer associated transcript 2 (CCAT2) lncRNA in CC and elucidate its possible mechanisms of action. The expression of CCAT2, the miR-493-5p microRNA (miRNA), and mRNA was detected using qRT-PCR. Cell viability, proliferation, and migration and invasion were determined using the MTT, colony formation, and transwell assays, respectively. The interactions between miR-493-5p and CCAT2 or cAMP response element-binding protein 1 (CREB1) were verified using the luciferase and RNA pull-down assays. The effects of CCAT2 knockdown on in vivo tumor growth were determined using tumor xenografts and immunohistochemistry assays. The expression of CCAT2 was upregulated in CC cells and tissues. However, the knockdown of CCAT2 inhibited the proliferation and epithelial-mesenchymal transition (EMT) of CC cells in vitro and suppressed tumor growth in vivo. Mechanistically, CCAT2 functions as a competing endogenous RNA (ceRNA) to upregulate the expression of CREB1 by binding to miR-493-5p. The overexpression of CREB1 or downregulation of miR-493-5p antagonized the effect of CCAT2 knockdown on the proliferation and EMT of CC cells. The knockdown of CCAT2 suppressed the aggressiveness of CC via the miR-493-5p/CREB1 axis. Therefore, CCAT2 is likely to be a promising therapeutic target for CC.
研究领域:
肿瘤学
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分子生物学
·
细胞生物学
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PubMed
高影响因子
Biochemistry Research International
·
2020
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IF 0
Yaoyv Xiang,Yanlin Li,Lingjian Yang,Yinghong He,Di Jia,Xidan Hu
Osteoarthritis (OA) is a chronic joint function disorder with characteristics of chondrocytes reduction and extracellular matrix (ECM) components destruction. MicroRNAs (miRNAs) and the SDF-1/CXCR4 axis are essential factors of chondrocyte apoptosis and ECM degeneration. However, very few studies have investigated the correlation between miRNAs and the SDF-1/CXCR4 axis in osteoarthritis so far. Here, through miRNAs microarray and bioinformatics analyses, we identified miR-142-5p as a CXCR4-targeted and dramatically downregulated miRNA in cartilage from OA patients, as well as in SDF-1-induced OA chondrocytes in vitro. In SDF-1-treated primary human OA chondrocytes that were transfected with a miR-142-5p mimic or inhibitor, the expression of CXCR4 was found to be inversely correlated with the expression of miR-142-5p. The dual luciferase reporter assay further verified the target relationship between miR-142-5p and CXCR4. Overexpression of miR-142-5p alleviated OA pathology by suppressing chondrocyte apoptosis, even in CXCR4 overexpressed OA chondrocytes. This was associated with decreased cartilage matrix degradation, reduced cartilage inflammation, and inactivated MAPK signaling pathway. Our study suggests that upregulated expression of CXCR4-targeted miR-142-5p can inhibit apoptosis, inflammation, and matrix catabolism and inactivate the MAPK signaling pathway in OA chondrocytes. Our work provides important insight into targeting miR-142-5p and the SDF-1/CXCR4 axis in OA therapy.
研究领域:
分子生物学
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风湿病学
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骨科学
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PubMed
BIOELECTROCHEMISTRY
·
2026
·
IF 4.9
Lan Jiang,Lina Zou,Jing Dong,Ping Zhang,Jian Zhou,Hao Jiang,Huanyu Ren,Huiru Niu,Hao Liao,Xiaojing Zhang,Shanshan An,Fei Ren,Xiuhong Ge,Feiyan Yang,Lang Cheng,Hongzhi Pan,Shengzhong Rong,Hongkun Ma
Non-coding RNAs are critical tumor biomarkers, rendering innovations in their detection technologies highly significant. To overcome the limitations of conventional single-signal electrochemical biosensors (susceptibility to interference, insufficient stability), this study developed a novel ratiometric electrochemical DNA biosensor with dual-signal amplification. Its core novelty is the rational integration of two synergistic nanomaterials: Ferrocene-functionalized MgAl-layered double hydroxide (Fc@MgAl-LDH, as stable immobilization matrix and internal reference signal source) and AuNPs@Fc@MgAl-LDH coupled with MB-cDNA@AuNPs, which construct a cascaded signal amplification pathway. The sensing mechanism involves two-stage amplification: Primary by high-loading-capacity AuNPs@Fc@MgAl-LDH, secondary by MB-cDNA@AuNPs hybridizing with electrode-immobilized microRNA-21 (miRNA-21). The ratiometric signal ( I MB / I Fc ) enables built-in correction, boosting detection accuracy and reproducibility. After systematic optimization, the sensor has a wide linear range (1.0 fM–0.1 nM) and an ultra-low LOD of 0.757 fM, eliminating RNA extraction/enzymatic amplification to simplify operation. Serum validation shows high accuracy (95.3%–101.4% recovery), indicating strong clinical diagnostic application potential.
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高影响因子
ARCHIVES OF DERMATOLOGICAL RESEARCH
·
2025
·Vol. 317
·期 1
·1-16
·
IF 2.1
Lin Ling,Li Wei,Gao Xinjing,Li Qian,Zhou Xin,Liu Weiyu,Zhong Xuelian,Yang Yunqing,Zhang Xibao,Luo Quan
Psoriasis represents a chronic inflammatory skin disease occurring globally. We investigated the role of plasma exosomal microRNA (miR)- 148a- 3p and its target gene Bim in psoriasis. Plasma exosomes (Exos) and CD4 + T cells were extracted from psoriatic patients. miR- 148a- 3p expression in Exos and CD4 + T cells of psoriatic patients, the proportions of CD4 + T cell subsets, and the contents of anti-inflammatory/pro-inflammatory factors were determined by RT-qPCR, flow cytometry and ELISA. The correlation between plasma exosomal miR- 148a- 3p and CD4 + T cell subsets in psoriatic patients was analyzed by Pearson’s analysis. The psoriatic mouse was treated with Exos/antagomir miR- 148a- 3p. Histopathological changes in the skin were observed. The CD4 + T cell subset levels, serum cytokine contents and miR- 148a- 3p expression in the blood were measured. The miR- 148a- 3p-Bim targeted binding relationship was predicted and verified by Starbase database and dual-luciferase assay. The Bim expression in psoriatic mice was determined. Psoriatic patients had highly-expressed miR- 148a- 3p in both Exos and CD4 + T cells, and abnormal CD4 + T cell subsets and cytokine levels. Plasma exosomal miR- 148a- 3p was correlated with the CD4 + T cell subsets in psoriatic patients. Exos caused down-regulated miR- 148a- 3p level in skin tissues of mouse, regulated CD4 + T cell function and aggravated the symptoms in psoriatic mice. miR- 148a- 3p repression partially reversed the role of Exos in CD4 + T cell function and psoriasis-like symptoms. Exos-carried miR- 148a- 3p targeted Bim in CD4 + T cells of psoriatic mice. Plasma exosomal miR- 148a- 3p targeted Bim to affect the dysfunction of CD4 + T cells in psoriatic mice, thereby aggravating the psoriasis-like symptoms.
研究领域:
皮肤病学
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分子生物学
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免疫学
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高影响因子
JOURNAL OF FLUORESCENCE
·
2025
·
IF 3.1
Xue Jinxiong,Huang Jingping,Lin Xue,Pan Wuyuan,Ke Fang
MicroRNA-183-5p is a clinically relevant biomarker, yet its detection is hindered by short sequence length, high homology, and low abundance in complex biological fluids. We report a hydrothermally synthesized nickel chloride-decorated graphitic carbon nitride (NiCl 2 @g-C 3 N 4 ) with intrinsic peroxidase-like and fluorescence activities for label-free dual-mode detection. Characterized by multiple spectroscopic and microscopic techniques, the nanocomposite enables quantitative analysis via π-π stacking and van der Waals interactions between the probe and target, eliminating the need for enzymes or fluorescent labels. Under optimal conditions, using the formula LOD = 3Sb/b, the detection limit was calculated as 0.69. This material with broad linear ranges, excellent selectivity, month-long stability, and recovery rates of 97.7-101.6% in spiked serum. This facile and cost-effective platform offers a promising tool for sensitive miRNA quantification in clinical diagnostics.
研究领域:
纳米材料
·
生物传感器
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临床诊断
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microRNA检测
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分析化学
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PubMed
高影响因子
ACS Nano
·
2025
·
IF 16
Xiaohui Zhan,Yujia Jiang,Zhiyu Li,Xiaoqin Hu,Fang Lan,Binwu Ying,Yao Wu
The screening and monitoring of microRNAs as cancer molecular biomarkers is clinically significant, but traditional methods lack sufficient sensitivity, accuracy, and convenience. The CRISPR-colorimetric lateral flow assay (CLFA) integration offers a promising and efficient solution; however, cumbersome preamplification and poor quantification hinder clinical adoption. In this study, we developed a one-step isothermal CRISPR-Cas cascaded sensing system that is preamplification-free. At its core is a designed and selected split DNA tetrahedron activator, employing spatiotemporal-hierarchy mechanisms to precisely bidirectionally drive the kinetics of two Cas enzymes, accelerating the activation of Cas13a while delaying the initiation of Cas12a, to achieve optimal balance. This system enables ultrasensitive, single-step, single-tube, and rapid detection of a cervical cancer relative biomarker, microRNA-21, achieving a limit of detection of 38 aM with a broad linear range. The CRISPR system is further integrated with CLFA enhanced by ultrathin platinum-protected gold nanolabels (Au@Pt, also named Au@s-Pt), along with a smartphone equipped with dual convolutional neural network models (YOLO v5 and MobileNet v3), enabling more precise, rapid quantification of target miRNA. Using this integrated platform, miRNA-21 levels in cervical cancer and precancerous samples can be accurately quantified with approximately 30 min at low cost and without the need for large, sophisticated instruments, with results showing good concordance with quantitative real-time polymerase chain reaction. This platform provides an efficient, highly sensitive, user-friendly, and quantifiable point-of-care testing solution.
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Small
·
2018
·Vol. 14
·期 40
·1802292
·
IF 9.598
Keying Zhang,Shuting Song,Shan Huang,Lin Yang,Qianhao Min,Xingcai Wu,Feng Lu,Jun-Jie Zhu
Intracellular microRNAs imaging based on upconversion nanoprobes has great potential in cancer diagnostics and treatments. However, the relatively low detection sensitivity limits their application. Herein, a lock-like DNA (LLD) generated by a hairpin DNA (H1) hybridizing with a bolt DNA (bDNA) sequence is designed, which is used to program upconversion nanoparticles (UCNPs, NaYF 4 @NaYF 4 :Yb, Er@NaYF 4 ) and gold nanoparticles (AuNPs). The upconversion emission is quenched through luminescence resonance energy transfer (LRET). The multiple LLD can be repeatedly opened by one copy of target microRNA under the aid of fuel hairpin DNA strands (H2) to trigger disassembly of AuNPs from the UCNP, resulting in the lighting up of UCNPs with a high detection signal gain. This strategy is verified using microRNA-21 as model. The expression level of microRNA-21 in various cells lines can be sensitively measured in vitro, meanwhile cancer cells and normal cells can be easily and accurately distinguished by intracellular microRNA-21 imaging via the nanoprobes. The detection limit is about 1000 times lower than that of the previously reported upconversion nanoprobes without signal amplification. This is the first time a nonenzymatic signal amplification method has been combined with UCNPs for imaging intracellular microRNAs, which has great potential for cancer diagnosis.
研究领域:
纳米技术
·
分子生物学
·
癌症研究
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PubMed
高影响因子
MICROCHIMICA ACTA
·
2018
·Vol. 185
·期 3
·1-9
·
IF 5.705
Wang Wei,Jayachandran Silambarasan,Li Mengru,Xu Shenghao,Luo Xiliang
An electrochemical biosensor for the detection of microRNA was prepared via chemical grafting of a Methylene Blue labeled reporter (MB-Rep) duplex onto a nanostructured surface that was obtained by electrodeposition of cobalt oxide and poly(o-phenylenediamine). This is followed by the attachment of hyaluronic acid and gold nanoclusters. In the presence of the target (microRNA), the probe-target duplex and the MB-Rep hairpin are formed. These will displace the labeled reporter from the sensor surface, and this results in a decrease of the amperometric signal for MB at a typical working voltage of −0.28 V (vs. Ag/AgCl). The electrode modified with hyaluronic acid possesses a large electroactive surface area and an excellent antifouling property. This makes it useful for ultrasensitive quantitation of microRNA even in complex biological media. The sensor has a linear response in the 100 f. to 0.1 μM microRNA concentration range, and a 33.3 f. detection limit. It was successfully applied to the determination of microRNA in cancer cells.
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研究领域:
生物化学
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电化学
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纳米技术
·
癌症诊断
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DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2020
·
IF 7.1
Fenghe Duan,Chuanpan Guo,Mengyao Hu,Yingpan Song,Minghua Wang,Linghao He,Zhihong Zhang,Riccardo Pettinari,Liming Zhou
A promising nanocarrier of the complementary DNA (cDNA) toward miRNA-155 was synthesized on the basis of the nanohybrid type of Ti 3 C 2 T x MXene nanosheets decorated using iron phthalocyanine quantum dots (FePc QDs) (denoted as Ti 3 C 2 T x @FePcQDs) to construct a novel ultrasensitive impedimetric aptasensing system for microRNA-155 (miRNA-155). Owing to π–π* stacking interaction between Ti 3 C 2 T x nanosheets and FePc QDs, the homogeneous nanostructure of the Ti 3 C 2 T x @FePcQD nanohybrid that comprises mixed-valence states (Ti 2+ /Ti 3+ and Fe 2+ /Fe 3+ ), multicomponent (Ti-O and Ti-C), and various N-related groups was achieved. The constructed Ti 3 C 2 T x @FePcQDs-based aptasensor displayed an ultrahigh sensitivity for detecting miRNA-155 with a low detection limit of 4.3 aM (S/N = 3) within the miRNA-155 concentration ranging from 0.01 fM to 10 pM. Compared with the individual component-based aptasensors and other reported miRNA-155 aptasensors, the proposed impedimetric aptasensing system exhibited substantial merits of a feasible preparation process, non-use of labels or electrochemical indicators, fast response time, and comprehensive sensing performances for detecting miRNA-155. This strategy for determining miRNAs can extensively be applied as the platform for anchoring other kinds of aptamers in detecting diverse targets, thus indicating its great potential application for the early diagnosis of cancer biomarkers.
研究领域:
纳米技术
·
生物传感器
·
分析化学
·
分子生物学
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癌症研究
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高影响因子
INTERNATIONAL JOURNAL OF MOLECULAR MEDICINE
·
2018
·Vol. 43
·期 1
·358-370
·
IF 2.784
Aifeng Zhang,Guoyun Wang,Lihua Jia,Tao Su,Lili Zhang
Endometriosis (Ems) is a condition that refers to the ectopic implantation and growth of endometrial tissue outside the uterine cavity. The aim of the present study was to investigate the role of microRNA‑138 (miR‑138) in Ems and the possible underlying mechanism. Flow cytometry was measured CD11b level, cell proliferation was measured using MTT assay and lactate dehydrogenase (LDH) assays was analyzed using LDH activity kits. Cell apoptosis was measured using Annexin V‑FITC/PI double staining apoptosis detection kit and DAPI assays. ELISA assay and western blot analysis were used to measure protein expression determination. It was first observed that miR‑138 expression was markedly downregulated and the CD11b level was reduced in Ems mice compared with the control group. Subsequently, miR‑138 expression was downregulated in the uterine endothelial cells co‑cultured with THP‑1 cells, which resulted in decreased apoptosis and increased inflammation in the uterine endothelial cells. By contrast, upregulation of miR‑138 by mimic transfection increased the proliferation and reduced inflammation in uterine endothelial cells. In addition, in the co‑culture of uterine endothelial and THP‑1 cells, downregulation of miR‑138 induced the expression of nuclear factor (NF)‑κB and vascular endothelial growth factor (VEGF) proteins in THP‑1 cells. Furthermore, treatment with an NF‑κB inhibitor and downregulation of miR‑138 in the co‑culture of uterine endothelial and THP‑1 cells reduced inflammation. VEGF inhibitor treatment and downregulation of miR‑138 in this cell co‑culture promoted the proliferation of uterine endothelial cells. These results suggested that uterine endothelial cells promoted miR‑138 to induce exosome‑mediated inflammation and apoptosis in Ems through the VEGF/NF‑κB signaling pathway.
研究领域:
妇科学
·
分子生物学
·
细胞生物学
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PubMed
高影响因子
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