Analytical Methods
·
2024
·Vol. 16
·期 25
·4116-4123
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IF 2.7
Yaxing Xie,Yulei Hou,Yang Yu,Jianhong Zhang,Jinyan Long,Mengqi Chen,Xueqing Lang,Xiaolan Yang,Hui Chen
MicroRNAs (miRNAs) have been involved in many biological processes and are regarded as promising biomarkers. The short sequence, low abundance and highly homologous interference sequences greatly hinder the accurate detection of miRNAs. Here, a cascade branch migration-triggered strand displacement amplification (CBM-TSDA) strategy was developed for the first time for specific and sensitive detection of miRNA-155 (miR-155). In the presence of target miR-155, the CBM was initiated and two Y-shaped probes were eventually produced. Next, the Y-shaped probes were transformed into three-way junction (3WJ) structures and triggered the SDA to produce a large number of G-quadruplex (G4) structures. Finally, the increased fluorescence signal of G4/Thioflavin T (ThT) was used to quantify miR-155. Meanwhile, the colorimetric responses of the G4-hemin DNAzyme could be used as supplementary detection to obtain a dual-mode signal readout. This detection strategy showed high detection sensitivity, and the limit of detection was 0.28 pM in the fluorescence detection mode and 0.34 pM in the colorimetric detection mode. Notably, it showed high detection specificity, being able to discriminate the single-base mutations of the target with a high discrimination factor. The strategy also possessed excellent capacity for miR-155 detection in cell lysates and real human blood samples. The developed strategy provides a promising detection platform for miRNA, which may be applied to early clinical diagnosis.
研究领域:
生物化学
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分子生物学
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临床诊断
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DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2022
·
IF 8.008
Fang Yang,Shunmei Li,Xia Li,Ruo Yuan,Yun Xiang
Accurate and specific imaging of low-abundance intracellular microRNAs (miRNAs) is crucial for monitoring cellular processes and disease diagnosis. Despite the fact that nucleic acid amplification technologies have shown great advantages for the detection of trace targets, their live cell imaging applications remain a major challenge because of the insufficient stability and slow reaction kinetics of the probes in cellular delivery and imaging. Herein, we demonstrate the synthesis of DNA-cross-linked polymeric lighting-up nanogels (DPLNs) through the DNA hairpin-based hybridization chain reaction within nanoscale-confined space for monitoring and imaging live cell miRNA-21 with high sensitivity. Cascaded catalytic hairpin assembly of two hairpin signal probes confined in the DPLNs can be triggered by the target miRNA, causing substantially amplified fluorescence resonance energy transfer signals with accelerated reaction kinetics. Moreover, the DPLNs show low cytotoxicity and highly enhanced nuclease resistance and can be successfully delivered into live cells for imaging low levels of miRNA-21. In addition, the DPLN probes can be readily tuned by specific sequences for monitoring various molecular targets in live cells for important biological and biomedical applications.
研究领域:
生物化学
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分子生物学
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医学诊断
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DOI
PubMed
高影响因子
PHYTOMEDICINE
·
2021
·
IF 5.34
Zheng Niu,Min Fu,Yuan Li,Huanhuan Ren,Xuanyu Zhang,Li Yao
Background Pulmonary vascular remodeling is the key pathological feature of pulmonary arterial hypertension (PAH) characterized by a pattern of lipid-related insulin resistance(IR), hormonal derangements and metabolic reprogramming. Our previous studies have demonstrated osthole as natural coumarin compound derived from traditional Chinese medicine is a promising agent for the treatment of pulmonary vascular remodeling in PAH. Purpose The present study sought to delineate lipid metabolic modulatory mechanism of osthole against pulmonary vascular remodeling by employing an interdisciplinary strategy. Methods Rat model with PAH induced with MCT and PASMCs proliferation model induced with PDGF-BB were established in this study. Serum and lung tissues were used to lipid-related IR, hormone related indexes, pulmonary vascular remodeling analysis. Then, lipid metabolic gene, key enzymes , metabolites and cell proliferation indexes were examined to investigate metabolic regulatory mechanism in vivo and vitro model of PAH. Results Osthole significantly showed improvement of lipid-related IR and hormone dysregulation in rats with PAH evidenced by elevating testosterone, androgen receptor and cyclic guanosine monophosphate (cGMP), inhibiting phosphodiesterase-5(PDE-5), modulating lipid-related IR indexes total cholesterol (TC), high density lipoprotein cholesterol (HDL-C), triglyceride (TG)/HDL-C ratio. Additionally, osthole limited key metabolic gene and enzymes to inhibit accumulation of decadienyl-l-carnitine in lipid metabolism , thus to promote oxidative phosphorylation and ATP production through inhibition of miRNA-22–3p, fatty acid translocase (CD36), fatty acid synthase (FAS), phospholipase A2 (PLA2), carnitine palmitoyltransferase 1A (CPT1A), hexokinase 2 (HK2), activation of metabolic switch isocitrate dehydrogenase 3α (IDH3α), NADH dehydrogenase 1 (ND1). We found for the first time miRNA-22–3p modulated PASMCs proliferation and vascular remodeling by regulating lipid metabolism reprogramming. Those modifications uncovered therapeutic mechanism of osthole against pulmonary vascular remodeling. Conclusion Our findings revealed the function of miRNA-22–3p in PASMCs and demonstrated a novel mechanism that miRNA-22–3p as a regulator can be targeted by osthole to greatly restore dysregulated lipid metabolism thus to alleviate pulmonary vascular remodeling in PAH, which provides novel insight into the potential therapeutic target for PAH, further highlights the development potential of osthole derived new drug against PAH.
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DOI
PubMed
高影响因子
TISSUE & CELL
·
2020
·
IF 1.837
Haocong Zhang,Meihui Piao,Mingming Guo,Lingzhi Meng,Hailong Yu
The recovery of spinal cord injury (SCI) involves multiple factors, of which miRNAs take an important part. In this study, we evaluated the function of microRNA-211−5p (miR-211−5p) on SCI in a rat model. SCI model was established using modified Allen's weight-drop method and Basso-Bcattie-Bresnahan score was applied to assess the locomotor function. MiR-211−5p agomir was utilized to increase miR-211−5p expression and endoplasmic reticulum (ER) stress inhibitor, 4-PBA (4-phenylbutyric acid), was utilized to suppress ER stress. Neuron apoptosis and the expressions of miR-211−5p, activating transcription factor 6 (ATF6), apoptosis-related proteins, pro-inflammatory cytokines and endoplasmic reticulum stress-related proteins were detected. Dual luciferase reporter gene assay was performed to verify the binding between miR-211−5p and ATF6. The results showed that miR-211−5p directly targeted ATF6. MiR-211−5p was down-regulated and ATF6 was up-regulated in SCI rats. Both interferences with miR-211−5p agomir and 4-PBA effectively attenuated neuron apoptosis and reversed the expressions of apoptosis, inflammation and endoplasmic reticulum stress-related molecules post SCI in rats. These findings demonstrated that miR-211−5p could effectively alleviate SCI-induced neuron apoptosis and inflammation via directly targeting ATF-6 and regulating ER stress.
研究领域:
神经科学
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分子生物学
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药理学
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DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
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2021
·
IF 7.46
Zilian Jiang,Jiao Jiao,Jianjun Li,Hongfang Zhang,Jianbin Zheng
MicroRNAs are novel and reliable biomarkers for early cancer diagnosing. However, accurate detection of the ultralow amount of a specific microRNA remains a big challenge. Herein, an electrochemical biosensing platform based on bivalent recognition-induced nanoparticle blocking in nanochannels for signal amplification is developed. Bivalent recognition is realized by two oligonucleotides, individually containing the complementary sequence for opposite ends of the target microRNA. One oligonucleotide is anchored into the alumina nanochannels and the other is bonded onto the SiO 2 nanoparticles. The presence of the target microRNA increases the blockage effect of the nanochannels and which is greatly amplified by bringing the nanoparticles into the nanochannels. Methylene blue (MB) is chosen as the reporter of the blockage effect. The amount of MB fluxing through the nanochannels is monitored at a working electrode functionalized with MoS 2 nanoplates and treated by the electrochemical method to further amplifying the signal. The reductive current of MB is quantitatively dependent on the concentration of microRNA. Using microRNA-21 as the model target, the electrochemical signal exhibited a linear dependence on the concentration of the target in a wide range of 100 aM to 1 nM. The platform could be readily extended to other nucleic acids.
研究领域:
生物技术
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医学诊断
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纳米科技
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产品关联
DOI
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2020
·
IF 7.1
Jia Tang,Yuxin Xing,Zhenqiang Wang,Mengnan Yang,Jixi Zhang,Kaiyong Cai
The design and interfacial engineering of DNA functionalized nanosensors with spatially separated but functionally integrated detection units holds a great promise in optical biosensors. In this study, we developed a bi-component hybrid of Janus nanoquencher particles with immobilized enzymes for fluorescence-based “off-on” detection in solution. Specifically, the hybrids consisting of a mesoporous polydopamine nanoparticle (MPDA) side (∼200 nm) asymmetrically attached with a gold (Au) particle side (∼30 nm) were generated through electrostatic repulsion-controlled growth. By regioselective binding and modification of the exposed side of the particles, fluorescent DNA probes and duplex-specific nuclease (DSN)/T7 exonuclease (T7) were subcompartmentalized on the MPDA and Au side, respectively, to realize substrate recognition and target recycling via the formation and hydrolysis of DNA hybrids. By using microRNAs (miRNAs) as a model target, the developed nanosensor realized a sensitive detection with a low limit of 32 fM (in a linear range from 20 fM–500 fM). Additionally, a high selectivity capable of discriminating a single-base mismatch, as well as a long-term detection stability were realized by the asymmetric interface designs and enzyme immobilization. The methodology is promising in the design and development of DNA probe-loaded hybrid nanosensors.
研究领域:
生物传感器
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纳米技术
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分子生物学
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分析化学
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DOI
高影响因子
MICROCHIMICA ACTA
·
2019
·Vol. 186
·期 7
·1-8
·
IF 5.479
Chen Donghua,Wen Siming,Peng Rulin,Gong Qingsong,Fei Junjie,Fu Zhuo,Weng Chao,Liu Minna
Mesoporous silica nanospheres (MSNs) are used in a triple signal amplification chemiluminescent (CL) assay for microRNA-21. It is based on (a) the synergistic amplification via loading and controlled-release of signal reagents by MSNs, (b) target recycling amplification, and (c) the enhancement effect of graphene oxide quantum dots (GOQD). CL is generated by the bis(2,4,6-trichlorophenyl) oxalate (TCPO) and H 2 O 2 reaction in the presence of the fluorophore rhodamine B (RB). RB is firstly loaded into the pores of MSNs modified with amino groupsand coupled with ssDNA. Then, the pores are capped by GOQD. Upon the addition of microRNA-21 into the system, the designed ssDNA assumes a double stranded structure. With the aid of duplex-specific nuclease, the double strand structure is cleaved and the free microRNA-21 enters into the next cycling process to combine with other ssDNA forming double strand structures. After several cycling process, amounts of GOQDs departing from the surface of MSNs cause the opening of the pores of MSNs and the release of RB causes the CL of TCPO-H 2 O 2 reaction system. Free GOQDs can lead to a further CL enhancement. By this method, even a low amount of microRNA-21 leads to a large number of released RB molecules and triggers high-intensity CL. The method was applied in an assay where the CL signal increases linearly with the logarithm of the microRNA-21 concentration in the range of 0.005–50 pmol L −1 and the detection limit is 1.7 fmol L −1 (at 3σ).
Schematic presentation of a triple signal amplification chemiluminescence (CL) analysis platform based on rodamine B (RB) loading and controlled release, target recycling amplification and graphene oxide quantum dots (GOQD) as the enhancer for analysis of microRNA-21 in human serum.
研究领域:
癌症研究
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分析化学
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纳米技术
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生物标志物检测
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DOI
PubMed
高影响因子
ANALYTICA CHIMICA ACTA
·
2019
·
IF 5.256
Lina Wu,Yingfei Wang,Rong He,Yue Zhang,Yuling He,Chao Wang,Zhenda Lu,Ying Liu,Huangxian Ju
Despite the great success of fluorescence sensing array application in bioanalysis , the photo-quenching problem accompanied with high surface fluorophore density has become the bottleneck for its further sensitivity improvement. Herein, a fluorescence polyethylene glycol (PEG) hydrogel array was developed for highly sensitive microRNA detection based on interfacial cation exchange amplification coupled with DNA hybridization chain reaction (HCR). The carboxylate PEG hydrogel array was fabricated on glass slide via photo polymerization and modified with miRNA capture probe, target miRNA binding triggered HCR in hydrogel with biotin labelled DNA probes to generate numerous DNA polymer chains where biotinylated CdS QDs were subsequently conjugated with a streptavidin bridge. Interfacial cation exchange amplification was triggered in hydrogel upon the introduction of Ag + and Rhod-5N, and abundant Cd 2+ was released from CdS to bind with Rhod-5N for substantial fluorescence enhancement. The aqueous-like environment of hydrogel eliminated the fluorescence quenching and simplified experiment process by performing cation exchange reaction at interface with direct result. The linear range for model target miRNA-21 was 1 fM to 500 pM with a detection limit of 0.835 fM. Taking advantage of the nonfouling property of PEG hydrogel, direct quantification of miRNA-21 was achieved from crude cancer cell lysates with a detection limit down to 10 equivalent cells. The expressions of circulating miRNA-21 in clinical serum samples were also assessed with comparable results from RT-PCR. The developed hydrogel array provides a universal platform for highly sensitive fluorescence sensing, and would benefit clinical non-invasive disease diagnosis.
研究领域:
生物化学
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生物技术
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分析化学
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分子生物学
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临床诊断
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产品关联
DOI
PubMed
高影响因子
RSC Advances
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2012
·Vol. 2
·期 18
·7140-7145
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IF 0
Yunlei Zhou,Mo Wang,Xiaomeng Meng,Huanshun Yin,Shiyun Ai
A novel signal amplified electrochemical microRNA (miRNA) biosensor was developed here using hemin-G-quadruplex as signal unit. A hairpin structure DNA probe (43 bases) was immobilized on the gold nanoparticles (AuNPs) modified gold electrode surface with the segment at its 3′-end complementary to miRNA-21 (22 bases) and the segment at its 5′-end complementary to capture DNA (21 bases). Upon hybridization with the target miRNA-21, the hairpin structure was unfolded, which was further hybridized with capture DNA loaded on AuNPs. The AuNP contained two kinds of DNA; one was complementary to the hairpin structure DNA probe, while the other was aptamer for hemin. The electrochemical signal of hemin self-assembled in the center of G-quadruplex structure of the aptamer was measured using chronoamperometry. The miRNA-21 target was analyzed with a detection limit of 3.96 pM. Using isolated total RNA from human gastric carcinoma BGC-823 cells, human breast adenocarcinoma MCF-7 cells, human hepatocarcinoma HepG2 cells and human colon cancer HT-29 cells, the assay detected specifically miRNA-21 and over-expression of oncogene miRNA-21 was demonstrated.
研究领域:
生物化学
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生物技术
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分析化学
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癌症研究
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产品关联
DOI
SENSORS AND ACTUATORS B-CHEMICAL
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2018
·
IF 5.667
Jiasheng Wang,Ni Hui
A simple electrochemical patterning strategy has been developed for the construction of ultrasensitive biosensors based on polyethylene glycol (PEG)-polypyrrole (PPy) nanowires substrate in order to supply improved antifouling performances. PPy nanowires array was produced through electrochemical polymerization of pyrrole at constant potential onto glassy carbon electrode (GCE) surface. PPy nanowires surfaces were decorated with 4-armed PEG molecules by electrochemical oxidation of amine groups offered by PEG end chains. The prepared PEG/PPy nanowires have integrated the excellent electrical conductivity of conducting polymer PPy nanowires with the good antifouling property of PEG. MicroRNAs (miRNAs) play very important roles in cancer development and a variety of diseases, which make them become promising biomarkers of the onset, prognosis and risk of diseases. Therefore the establishment of miRNA profiles for diseases and the detection of miRNAs in biological samples are critical milestones in diagnostics. An ultrasensitive electrochemical biosensor for miRNA can be effortlessly developed by the immobilization of DNA probes onto PEG/PPy nanowires. DNA/RNA hybridization was monitored by changes of methylene blue (MB) redox signal using differential pulse voltammetry (DPV) method. A wide linear range (0.10 pM ∼ 1.0 nM) to target miRNA was obtained by the fabricated biosensor, and miRNA mismatches can also be easily identified with satisfactory. Furthermore, the biosensor based on PEG/PPy nanowires can be extended to the development of any type of DNA-based biosensor.
研究领域:
生物传感
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分子诊断
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电化学
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材料科学
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DOI
高影响因子
PLoS One
·
2016
·Vol. 11
·期 3
·e0149257
·
IF 3.057
Chao Fang,Weihong Lu,Chengyan Li,Xi Peng,Yang Wang,Xiulan Huang,Zhihong Yao,Nali Cai,Yuge Huang,Xingliang Zhang,Jianxin Tan
Asthma is a common chronic respiratory disease. In a previous study, we found several circulating microRNA signatures associated with childhood asthma and selected miR-3162-3p for subsequent studies. Since the target proteins and underlying molecular mechanisms of miR-3162-3p in asthma etiopathogenesis are not well characterized, we designed this study to clarify its role. We employed bioinformatics and quantitative PCR methods as a first step to determine the target of miR-3162-3p, and we elucidated β-catenin. Luciferase assays and western blot analysis confirmed β-catenin as a direct target of miR-3162-3p as the 3’-untranslated region of β-catenin mRNA possesses a specific miR-3162-3p pairing site. The correlation between the expression levels of miR-3162-3p and β-catenin is confirmed by quantitative PCR and western blot studies in A549, Beas-2B and H1299 cell lines and OVA-induced asthma mouse model. Of note, upregulation of the endogenous miR-3162-3p level is concomitant with the reduction of β-catenin mRNA and protein expression levels. MiR-3162-3p antagomir treatment antagonizes the endogenous miR-3162-3p and effectively rescues the attenuation of endogenous β-catenin in OVA-induced asthmatic mice, which alleviates airway hyperresponsiveness and ameliorates airway inflammation. Collectively, our findings suggest a novel relationship between miR-3162-3p and β-catenin and clarify their mechanistic role in asthma etiopathogenesis.
研究领域:
分子生物学
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呼吸病学
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免疫学
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产品关联
DOI
PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
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2022
·
IF 12.545
Fu-Ting Wang,Yang-Yang Hou,Xuecai Tan,Ke-Jing Huang,Jing Xu,Ren Cai
A real-time self-powered biosensor is designed for ultrasensitive detection of microRNA-21 based on electrochemical energy device capacitor and target-induced recycling double amplification strategy, which greatly improves the output signal by converting a small number of targets into two glucose oxidase labeled output strand DNAs , and the squeezed-out output strand is recycled by the cathode to fix more signal [Ru(NH 3 ) 6 ] 3+ to further improve the detection signal. A digital multimeter (DMM) is connected to computer for real-time displaying the output signal of the self-powered biosensing system, which improves the accuracy of the sensing platform. The sensitivity of the proposed biosensor is 116.15 μA/pM for target microRNA-21, which is 32.26 times higher than that of pure EBFC (3.6 μA/pM). The target concentration is proportional to the open-circuit voltage value in a wide linear range of 0.1–10000 fM with a low detection limit of 0.04 fM (S/N = 3). The method shows high sensitivity and excellent selectivity , and can be applied to detect tumor marker microRNA-21 in biological matrix.
研究领域:
生物传感器
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电化学
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分子诊断
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癌症研究
·
纳米技术
✓ 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）