MICROCHEMICAL JOURNAL
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2024
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IF 4.9
Yanlian Niu,Zezhao Meng,Guofeng Qiu,Han Zhao,Shulan Liu,Nan Sun,Wei Qin
A high sensitive sandwich-type electrochemical biosensor utilizing capture probe (CP), target probe (TP), and signal probe (SP) was first developed for specific detection of microRNA-26a (miR-26a), a biomarker with diagnostic significance for atherosclerosis. In general, gold nanoparticles (Au NPs) were electrodeposited on to a glassy carbon electrode to enhance conductivity, followed by immobilization of 5 ′- thiolated CP through a gold-sulfur bond. Subsequently, the modified electrode was hybridized with the mixture of miR-26a and 3′-biotin-tagged SP in sequence. Furthermore, horseradish peroxidase-streptavidin (HRP) with potent reducing capability was finally grafted on the SP via the strong affinity between biotin and streptavidin. Additionally, the fabrication process of the miR-26a biosensor was thoroughly investigated. Based on the dual amplification mechanism facilitated by Au NPs and HRP-catalytic activity, the proposed biosensor demonstrated outstanding electrochemical properties, including good reproducibility and stability, as well as high sensitivity for miR-26a detection down to 0.88 fM with a linear range from 1 fM to 1 nM. The biosensor also displayed exact specificity, which could discriminate miR-26a from a range of base mismatched microRNAs. Moreover, it was successfully applied to the determination of miR-26a in serum samples. In conclusion, given the excellent analytical performance, this biosensor shows promise for potential use in clinical diagnostics for atherosclerosis.
研究领域:
生物化学
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纳米技术
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生物医学工程
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分析化学
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临床诊断
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DOI
高影响因子
MICROCHEMICAL JOURNAL
·
2024
·
IF 4.9
Yuanyi Wu,Yan Yang,Shihao Yang,Yang Ju,Yang Jiang,Yumei Xiao,Jialin He,Tian Liu,Leyao Tang,Kaiyong Wang,Yi Yang
Interfacial and interlayer interactions between layered double hydroxides (LDHs) and nucleic acids offer LDHs great potential to absorb and detect nucleic acids. Herein, we demonstrate a simple, rapid, and versatile fluorescent sensing platform based on two-dimensional LDHs and exonuclease I (Exo I) for the selective analysis of DNA and microRNA. ZnNiAl-LDHs were utilized as quenchers for FAM labelled on single-stranded DNA (ssDNA) probes via dynamic quenching with high fluorescence quenching ability. The quenching mechanism was explored by means of spectral analysis, characterization experiments, DNA adsorption and desorption studies, zeta potential analysis, and molecular dynamics simulations. The driving forces for the adsorption of LDHs toward fluorophore-labelled nucleic acids were confirmed to be electrostatic interaction, van der Waals force, and anion exchange for the first time. In order to distinguish the formed target/probe duplexes from FAM-ssDNA probes, Exo I was introduced to specifically hydrolyze FAM-ssDNA probes. The fabricated fluorescent sensing platform for nucleic acids detection shows a low detection limit of 0.08 nM. Finally, this sensor was effectively utilized for the detection of DNA and microRNA in actual biological and environmental samples, achieving recoveries ranging from 96.4 % to 105 %, indicating high accuracy. This work proposed a promising sensing platform for sequence-specific nucleic acids detection, and provides a new perspective on two-dimensional LDHs as structural supports and biomolecular reservoirs for biomedical applications.
研究领域:
生物技术
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纳米技术
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分析化学
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分子生物学
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高影响因子
MICROCHEMICAL JOURNAL
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2026
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IF 5.2
Yi Liang,Ke Huang,Jin-Wen Liu,Yumin Wang
Thioflavin T as split G-quadruplex fluorescent dye was researched. • A novel sensing method based on CoOOH nanoflakes and HCR was proposed. • Label-free and sensitive detection of miRNA-21 was successfully realized. To date, various fluorescent probes for G-quadruplex have been reported. However, the development of fluorescent dyes for split G-quadruplex formed on double-stranded DNA structure still remained largely unexplored. Herein, we have demonstrated that thioflavin T (ThT) was able to intercalate into split G-quadruplex sequence and the formed split G-quadruplex-ThT structure exhibited remarkable fluorescence enhancement. By integrating cobalt oxyhydroxide (CoOOH) nanoflakes and hybridization chain reaction (HCR), a sensitive and label-free biosensing strategy for miRNA detection was proposed. In our design, miRNA-21 was chosen as the model target and it was capable of triggering HCR with two split G-quadruplex sequence-contained hairpin probes, resulting in the generation of long chain-like HCR product. Unlike hairpin probes, the HCR product cannot be adsorbed onto CoOOH nanoflakes because of lower affinity. Consequently, in the presence of ThT, numerous split G-quadruplex-ThT structures on HCR product were formed and the target miRNA was converted into amplified and activatable fluorescence signal. Experimental results demonstrated that this novel sensing strategy can be applied for sensitive and selective detection of miRNA. Moreover, the analysis of miRNA in complex biological media was successfully realized, offering a robust, label-free and cost-effective platform for miRNA-related disease diagnosis and biomedical research. A novel fluorescence biosensing strategy based on cobalt oxyhydroxide (CoOOH) nanoflakes and hybridization chain reaction (HCR) was developed for sensitive and label-free detection of miRNA-21. Download: Download high-res image (161KB) Download: Download full-size image
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高影响因子
ACS Applied Nano Materials
·
2025
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IF 5.5
Yanci Zhang,Yujin Fan,Jiyuan Zhang,Yanxi Hu,Lili Duan,Zhanhui Wang,Yihong Wang,Jiansheng Cui,Liang Tian
The imbalanced expression of microRNA is related to many diseases and tumors, which can be used as a noninvasive biomarker to assist in the diagnosis, prognosis, and monitoring of cancer. A target conversion strategy-based electrochemical biosensor was developed by DNAzyme-cleavage dual cyclic signal amplification and the strand displacement reaction (SDR) to produce high electrochemical signals for microRNA-155 (miRNA-155) analysis. Thereinto, VSe2/multi-walled carbon nanotubes (MWCNTs) with high conductivity were applied as the electrode surface material to provide more active sites and accelerate electron transportation. Then, the excellent immobilization efficiency and large specific surface area of the ZnO@CuS nanoprobes were applied as a recognition molecule carrier. Furthermore, hexaammineruthenium(III) chloride (RuHex), with superior electrochemical redox activity, acted as the electroactive indicator by intercalating into double-helix DNA through electrostatic interaction, which produced high current responses in the presence of the target. Therefore, this miRNA-155 biosensing platform exhibited a relatively low limit of detection of 0.11 fM, ranging from 0.5 fM to 5 nM, and demonstrated excellent repeatability and stability. Hence, this platform appeared to possess favorable human serum detection capacity and be suitable for early cancer diagnostics.
研究领域:
生物医学工程
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分子诊断学
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纳米技术
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DOI
高影响因子
Acta Biomaterialia
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2025
·
IF 9.6
Chunfa Chen,Quan Li,Beinuo Wang,Qian Liu,Zi Wang,Bihang Zhang,Lu Wang,Yujuan Zou,Ziye Mou,Chengshan Ren,Zaichun You,Bin Wang,Zhi Xu,Hang Qian
Acute lung injury (ALI) is characterized by widespread inflammation and oxidative stress, leading to impaired gas exchange and significant morbidity. In this study, we propose a potential approach using a magnesium-free DNA self-assembly strategy to assemble a DNA nanotube that carries agmatine and microRNA-126b mimics (NT AGM -126). Agmatine not only reduces electrostatic repulsion between DNA helices, thereby facilitating the folding of the DNA nanotube, but also serves as a drug that inhibits iNOS signaling. The microRNA-126b mimics restore the downregulated microRNA-126b in macrophages and suppress inflammation by targeting high mobility group box 1 (HMGB1). Preliminary results indicated that agmatine can effectively facilitate the assembly of the DNA nanotube, improve serum stability, and enhance the cellular uptake efficiency of NT AGM -126. Further in vitro and in vivo results demonstrate that NT AGM -126 effectively reduces oxidative stress and inflammation by downregulating iNOS and HMGB1, providing a combined therapeutic effect in ALI. This study highlights the potential of agmatine-facilitated DNA nanostructures as a versatile drug delivery platform for treating inflammatory diseases, broadening the application of DNA nanotechnology in biomedical research. Statement of Significance This study introduces a promising therapeutic approach using a magnesium-free DNA self-assembly strategy to create a DNA nanotube (NT AGM -126) that carries agmatine and microRNA-126b mimics. The agmatine not only aids in the assembly and stability of the DNA nanotube but also inhibits iNOS signaling, while the microRNA-126b mimics restore downregulated microRNA-126b in macrophages and suppress inflammation by targeting HMGB1. Preliminary and further results demonstrate that NT AGM -126 effectively reduces oxidative stress and inflammation, providing a synergistic therapeutic effect in ALI. This study underscores the potential of agmatine-facilitated DNA nanostructures as a versatile drug delivery platform, broadening the application of DNA nanotechnology in the treatment of inflammatory diseases and advancing biomedical research.
研究领域:
生物医学工程
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分子生物学
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药学
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DOI
PubMed
高影响因子
Bioactive Materials
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2023
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IF 18.9
Hongyan Wei,Fenfang Li,Tiantian Xue,Haixia Wang,Enguo Ju,Mingqiang Li,Yu Tao
As the most abundant liver-specific microRNA, microRNA-122 (miR122) played a crucial role in the differentiation of stem cells into hepatocytes. However, highly efficient miR122 delivery still confronts challenges including poor cellular uptake and easy biodegradation. Herein, we for the first time demonstrated that the tetrahedral DNA (TDN) nanoplatform had great potential in inducing the differentiation of human mesenchymal stem cells (hMSCs) into functional hepatocyte-like cells (HLCs) by transferring the liver-specific miR122 to hMSCs efficiently without any extrinsic factors. As compared with miR122, miR122-functionalized TDN (TDN-miR122) could significantly up-regulate the protein expression levels of mature hepatocyte markers and hepatocyte-specific marker genes in hMSCs, indicating that TDN-miR122 could particularly activate the hepatocyte-specific properties of hMSCs for developing cell-based therapies in vitro . The transcriptomic analysis further indicated the potential mechanism that TDN-miR122 assisted hMSCs differentiated into functional HLCs. The TDN-miR122-hMSCs exhibited hepatic cell morphology phenotype, significantly up-regulated specific hepatocyte genes and hepatic biofunctions in comparison with the undifferentiated MSCs. Preclinical in vivo transplantation appeared that TDN-miR122-hMSCs in combination with or without TDN could efficiently rescue acute liver failure injury through hepatocyte function supplement, anti-apoptosis, cellular proliferation promotion, and anti-inflammatory. Collectively, our findings may provide a new and facile approach for hepatic differentiation of hMSCs for acute liver failure therapy. Further large animal model explorations are needed to study their potential in clinical translation in the future.
研究领域:
分子生物学
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干细胞研究
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肝病
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纳米技术
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再生医学
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DOI
PubMed
高影响因子
ARCHIVES OF DERMATOLOGICAL RESEARCH
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2024
·Vol. 317
·期 1
·1-12
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IF 1.8
Mu Yanan,Lian Chen,Chen Xinghui,Yang Xueying,Li Dongxia,Zhang Yi
Tumor-derived exosomes that transport MicroRNAs significantly influence cutaneous squamous cell carcinoma(CSCC) progression. However, the molecular mechanisms of exosomal MicroRNA-31 regulation of CSCC are mostly undefined. To determine whether a targeting relationship exists between MicroRNA-31 (miR-31) in CSCC-derived exosomes and the tumor suppressor RhoBTB1, and the regulatory effect of the relationship on tumor cells. Immunoblotting and quantitative PCR were used to measure miR-31 and RhoBTB1 levels in various cells and exosomes. Differential ultracentrifugation was used to isolate exosomes. MTT and Transwell assays were used to assess cell proliferation, migration, and invasion. Dual luciferase reporter assays were used to assess the direct interaction between miR-31 and the tumor suppressor RhoBTB1 in cutaneous squamous cell carcinoma (CSCC)-derived exosomes. Compared with a human skin keratinocyte cell line, in CSCC cell lines RhoBTB1 was downregulated and miR-31 levels were elevated. Exosomal miR-31 from CSCC cell lines directly targeted RhoBTB1 by binding to the 3’ UTR of RhoBTB1. This interaction suppressed expression of RhoBTB1 and enhanced CSCC cell proliferation, migration, and invasion. MicroRNA-31 in CSCC-derived exosomes can enhance CSCC cell proliferation, migration, and invasion by suppressing expression of RhoBTB1. This finding explains, in part, the molecular mechanism of CSCC. Investigative approaches focused on suppressing miR-31 or enhancing RhoBTB1 signaling pathways are promising avenues for developing targeted therapies for CSCC.
研究领域:
肿瘤学
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分子生物学
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细胞生物学
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DOI
PubMed
高影响因子
SPECTROCHIMICA ACTA PART A-MOLECULAR AND BIOMOLECULAR SPECTROSCOPY
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2022
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IF 4.831
Qingyou Cai,Fanfan Wang,Jingying Ge,Zhiguo Xu,Mei Li,Hui Xu,Hua Wang
Herein, we reported the G-wire-based self-quenched fluorescence probe and its application in ultrasensitive microRNA (miRNA) detection by combining with target-activated isothermal cascade amplification. The terminal-single-fluorescein (FAM)-labeled G-rich oligonucletides self-assembled into G-wire nanostructures (G-wires) with K + and Mg 2+ . Thereafter, the G-wires brought terminal-labeled FAM into close proximity, as a result, the self-quenched signal probe formed. Besides, when there was the target miRNA, target-activated isothermal cascade amplification converted miRNA into the copious trigger DNA. After hybridization between trigger DNA and the self-quenched probe, the G-wires were splited and forced the apart of proximate FAM, and then the self-quenched probe displayed an “on” mechanism. Therefore, the approach gave a limit of detection (LOM) of 0.82 aM to miRNA-21 and could be implemented within a wide linear range of 2 aM to 2 nM. This approach was able to distinguish the single-mismatched miRNA-21, which was selective and sensitive in detecting human spiked serum samples.
研究领域:
分子生物学
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生物化学
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纳米技术
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癌症诊断
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DOI
PubMed
高影响因子
MICROCHIMICA ACTA
·
2019
·Vol. 186
·期 8
·1-7
·
IF 5.479
Li Mei,Xu Xiong,Cai QingYou,Luo XuJian,Zhou ZhongGao,Xu GuoHai,Xie YongRong
A graphene oxide-based method has been developed for ultrasensitive and selective determination of microRNA-141 by means of rolling circle amplification (RCA) and exonuclease III (Exo III)-assisted recycling amplification. The method uses (a) a padlock probe with a hybrid sequence that is complementary to the target microRNA-141 at both the 5'- and the 3'-end, and (b) a long binding region of a signalling reporter strand. On addition of microRNA-141, it acts as the primer for triggering the RCA reaction following ligation. This results in the formation of a repeatedly concatenated sequence of the padlock probe. Subsequently, the RCA product hybridizes with the fluorescein-labelled signal strand to form the duplex DNA containing the blunt 3'-termini of signal strand. Addition of Exo III causes signal strand digestion and leads to RCA product recycling and liberation of fluorescein. Added graphene oxide does not absorb the fluorescein liberated because of the poor mutual interaction. Therefore, microRNA-141 can be quantified by measurement of the green fluorescence under excitation/emission wavelengths of 470/520 nm. The method has a 100 aM detection limit towards microRNA-141 and works in the wide range from 1 fM to 1 nM. It can discriminate even single-mismatched microRNA and shows good selectivity and sensitivity when applied to spiked human serum.
Schematic representation of a graphene oxide (GO)-based method for fluorometric determination of microRNA by using rolling circle amplification and exonuclease III (Exo III)-aided recycling amplification. With microRNA, the fluorescein-labelled signal strand becomes digested, and this genererates a fluorescent signal.
研究领域:
分子生物学
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生物化学
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纳米医学
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DOI
PubMed
高影响因子
TALANTA
·
2019
·
IF 4.916
Feng Zhang,Shuang Wang,Jian feng,Rong Zou,Ling Xiang,Changqun Cai
A molecular beacons (MBs) loaded on molybdenum disulfide (MoS 2 ) nanosheets as fluorescence probes for sensitive and versatile detection of microRNAs (miRNAs) through hybridization chain reaction (HCR) has been designed. MoS 2 was used as a adsorbent to capture the MBs and a selective fluorescence quencher to reduce the background signal. In the absence of miRNAs, HCR could not be triggered due to the stability of MB probes. The probes attached to the MoS 2 surface, efficiently quenching fluorescence of the G-quadruplex/Thioflavin T . However, the presence of target miRNAs triggers the HCR process to generate large amount of HCR products. Meanwhile, the HCR products of long nanowires chain with abundant G-quadruplexes could not be adsorbed on the surface of MoS 2 , and therefore detach from the MoS 2 . Consequently, Thioflavin T could be embedded in G-quadruplexes and produced strong fluorescence signal. This fluorescence emission signal could achieve detection of miRNA as low as 4.2 pM and a wide linear ranges from 0.1 to 100 nM. In addition, a versatile fluorescence probe has been developed for detection of miRNA-21 by changing the miRNA-recognition domain of MB. Thus, the fluorescent probe would be a potential alternative tool for biomedical research and clinical molecular diagnostics.
研究领域:
生物化学
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分子生物学
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纳米技术
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分析化学
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产品关联
DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2019
·
IF 6.35
Huan Gao,Junfang Zhang,Ying Liu,Wenwen Tu,Tianxiang Wei,Zhihui Dai
Development of sensitive detection methods for microRNAs has realistic application value for early clinical analysis and accurate diagnosis. In this study, a triple-helix molecular switch electrochemiluminescence resonance energy transfer (ECL-RET) nanoamplifier was designed to construct an electrochemiluminescence (ECL) biosensor for microRNA determination. The newly synthesized S-doped Lu2O3, which shows a 3 times better ECL performance than Lu2O3, was chosen as the donor in this ECL-RET system. Accordingly, Ag2S quantum dots (QDs) were used as the matched acceptor. They exhibited overlapping spectra and efficient energy transfer between each other. Furthermore, using a triple-helix switch structure with an improved quenching effect for signal amplification and a nano-DNA walker transformational system as a powerful method for target amplification, a supernanoamplifier was achieved. As a consequence, the proposed ECL biosensor for microRNA-141 detection exhibited good analytical performance with a low detection limit (16 aM). The sensor not only led to the development of a novel ECL-RET pair but also provided a strong nanoamplifier for biosensing platform construction, which may offer some new considerations regarding material design, signal amplification, and show promising application value in biomedical and clinical diagnosis.
研究领域:
生物传感技术
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纳米材料科学
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临床诊断
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产品关联
DOI
PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2016
·
IF 7.476
Hao Wu,Yaling Liu,Hongyong Wang,Jun Wu,Feifan Zhu,Pei Zou
In this study, a simple, label-free, and enzyme-free colorimetric biosensor has been developed for amplified detection of let-7a microRNA (miRNA) on the basis of dual signal amplification strategy. The sensing system mainly consists of four unlabeled hairpin probes termed H1, H2, H3, and H4. Upon sensing of the target miRNA, hairpin H1 is opened. Then hairpin H2 hybridizes with H1 forming H1–H2 duplex and frees the target miRNA that can be recycled to trigger another reaction cycle. In addition, the newly formed H1–H2 duplex hybridizes with hairpin H3, and this triggers the autonomous cross-opening of the two hairpins H3 and H4 through hybridization chain reaction. During this process, numerous split G-quadruplex structures are generated and further associate with cofactor hemin to form massive peroxidase-mimicking DNAzymes. The resulting DNAzymes catalyze the H 2 O 2 -mediated oxidation of colorless 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS 2− ) to the green-colored ABTS •− , inducing a remarkably amplified colorimetric signal. This newly developed sensing system exhibits high sensitivity toward miRNA with a detection limit of 7.4 fM and a large dynamic range of 6 orders of magnitude from 10 fM to 10 nM. Furthermore, it exhibits a good performance to discriminate single-base difference among the miRNA family members and holds a great potential for early diagnosis in gene-related diseases.
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
·
纳米技术
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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）