ANALYST
·
2023
·Vol. 148
·期 9
·2180-2188
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IF 4.2
Mengyao Li,Tingting Zhang,Yuzhong Zhang
In this study, an ultrasensitive electrochemical miRNA-21 biosensor is described. Manganese dioxide-gold nanoparticle (MnO2-Au NP) nanoconjugates were employed as sensing base materials, miRNA-21 was selected as a model analyte, and hybridization chain reaction (HCR) was employed to form long DNA concatemers using two different oligonucleotides with a complementary sequence. Thus, lots of biotin were loaded on DNA concatemers and one of them was labelled with biotin at its 3′ terminal. The biosensor was designed as follows: a sulfhydryl-hairpin probe (HP) was first dropped on the surface of the glassy carbon electrode (GCE) modified with MnO2-Au NP nanoconjugates (HP/MnO2-AuNPs/GCE). After it was treated with MCH, the modified electrode was hybridized with miRNA-21, resulting in the loop of HP being opened to form a vertical structure. Subsequently, the modified electrode (miRNA-21/HP/MCH/MnO2-AuNPs/GCE) was incubated with DNA concatemers to form a sandwich structure of HP-miRNA-21-DNA concatemers on the modified electrode surface. Finally, the streptavidin-HRP conjugates were linked to the sandwich structure by specific recognition interaction between biotin and avidin. Differential pulse voltammetry (DPV) was used to measure the electrochemical response of the biosensor in the phosphate-buffered solution (0.10 M PBS, pH 7.0) containing 2.0 mM hydroquinone (HQ) and 1.8 mM H2O2. As a result, a larger reductive signal was obtained at a potential of −0.17 V (vs. SCE). Various experimental conditions were optimized, including solution pH, incubation time, and the amount of DNA concatemers. Under optimal conditions, the biosensor showed good sensing performance, such as a wide linear response range (0.1 fM and 100 nM) and low detection limit (0.063 fM, at S/N = 3). Meanwhile, the biosensor can discriminate single base matched miRNA-21, indicating that the biosensor had good selectivity.
研究领域:
分析化学
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生物化学
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纳米技术
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分子生物学
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DOI
PubMed
高影响因子
Chemistry-An Asian Journal
·
2023
·
IF 4.1
Junyou Li,Hung-Wing Li
By combining exquisitely designed hairpins with the catalytic hairpin assembly to form tripedal DNA walkers driven by enzyme, we constructed a 3D DNA walker with accordingly complementary hairpins attached on gold nanoparticles (AuNPs) and sensitive fluorescence sensing system for the sensitive detection of target miRNA-21. The presence of miR-21 triggers the CHA among three hairpins, which lead to the formation of the tripedal DNA walkers. For the walking trajectories, FAM-labeled hairpins were attached to the surface of AuNPs, the fluorescence of which was initially quenched due to its close proximity to AuNPs. After the binding/cleaving/moving process of tripedal DNA walkers with HP4 driven by Exonuclease III (Exo III), a number of single-stranded DNAs (ssDNAs) will be released with FAM fluorescence recovered. Benefiting from the DNA walker and CHA cascade amplification, the proposed sensing strategy showed remarkable improvement in sensitivity with the LOD of 46 aM. Also, the system was practically applied in human serum samples, demonstrating satisfactory outcome. Owing to the precise design of the system, this method exhibited excellent specificity to distinguish miR-21 from its single-, double-mismatched sequences and non-complementary sequences, showing great versatility and potential for the biological analysis and early disease diagnosis.
研究领域:
生物技术
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纳米技术
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癌症诊断
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分子生物学
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分析化学
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DOI
PubMed
高影响因子
ACS Applied Materials & Interfaces
·
2022
·
IF 10.383
Yanli Zhu,Jikai Wang,Haitao Xie,Hailing Liu,Shuangquan Liu,Dongxiu He,Pengbing Mi,Suisui He,Jun Wang,Yiyang Sun
Sub-10 nm monodisperse alkaline-earth sulfide nanodots (ASNDs) with bright near-infrared (NIR)-excitation fluorescence and adjustable emission wavelength were prepared by a thermal decomposition method for the first time. The ASNDs exhibited high NIR-to-vis conversion efficiency and served as multicolor fluorescent labels in the proposed miR-224 assay. Targeted detection of the miR-224 level and single-nucleotide variation in miR-224 was carried out on a smartphone-based platform using a hybridization chain reaction (HCR) amplification strategy. In the presence of miR-224, the ASND-labeled HCR probes self-assembled on the surface of the diagnosis kits, generating strong fluorescent signals linearly proportional to miR-224 contents in the range of 10–2000 fM. Significantly, mutations in miR-224 led to the variation in the fluorescence intensity ratio in RGB channels. Simultaneously, evident changes of fluorescent brightness and color were easily visualized by the naked eye, which enabled on-site discrimination of miR-224 with different mutant loci. This work provides a novel preparation approach for ultrasmall NIR excitation sulfide nanodots and reveals the potential of the as-synthesized ASNDs in point-of-care (POC) nucleic acid testing. Further, it may provide a handheld platform for miRNA single-nucleotide polymorphism analysis.
研究领域:
生物化学
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纳米技术
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医学诊断
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分子生物学
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DOI
PubMed
高影响因子
Comparative Biochemistry and Physiology D-Genomics & Proteomics
·
2023
·
IF 3
Cheng Zhao,Sijin Wang,Yuxi Liu,Peng Chu,Bing Han,Xianhui Ning,Tao Wang,Shaowu Yin
Temperature is a critical factor that regulates the reproduction processes in teleost. However, the gonadal response mechanism to cold stress in fish remains largely unknown. In the present study, female zebrafish were exposed to different extents of low temperatures at 18 °C and 10 °C for 48 h. The ovarian histology was remarkably damaged after cold stress exposure. Integrated analysis of miRNA-mRNA was used to investigate the ovarian response to acute cold stress. A large number of mRNAs and miRNAs were altered by cold stress, which are involved in extensive biological processes . It is indicated that the signal transduction of MAPK and Calcium signaling pathway is highly engaged in zebrafish ovary to adapt to cold stress. The immune system was dysregulated by cold stress while the ovarian autophagy was activated. Remarkably increased gene number related to reproductive functions was identified in the cold stress at 10 °C compared to the control. The cold stress-induced dysregulated reproductive genes include star , hsd3b1 , hsd17b1 , inha , insl3 , amh , nanos1 and foxl2 . Combined with the dysregulated insulin, IGF and progesterone signaling, it is suggested that cold stress affects ovarian function in multiple aspects, including oocyte meiosis, folliculogenesis , final maturation and ovarian maintenance. On the other hand, the ovarian miRNA-mRNA regulatory network response to cold stress was also constructed. Overall, our result revealed the ovarian response to cold stress in zebrafish and provided insight into the fish adaptation mechanism to acute temperature change.
研究领域:
水生生物学
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生殖生物学
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分子生物学
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基因组学
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PubMed
高影响因子
ANALYTICA CHIMICA ACTA
·
2022
·
IF 6.911
Rui Zhang,Mingrui Zhang,Qiao Ding,Ruo Yuan,Yali Yuan
In this work, a novel label-free and signal-off electrochemical biosensor based on efficient target-trigger T-structure recycling with dual strand displacement amplification (T-DSDA) was constructed for sensitive detection of miRNA-155 that related to cardiovascular diseases. Compared with traditional SDA, the proposed strategy not only accomplished simple polymerization and shearing of dsDNA without continuous consumption of temple DNA , but also ingeniously designed two analogous sequences that produced double cleavage sites, thereby obtaining extensive output DNA for further cleaving Y-structure assembled on electrode with significantly improved target conversion efficiency. Since the redox probe methylene blue (MB) attach to Y-structure via electrostatic interaction and tessellation, an obviously decreased current with a typical signal-off mode could be acquired for sensing miRNA-155 in the assistant of DNAzyme cleavage recycling. As a result, the electrochemical biosensor realized the rapid and sensitive detection of miRNA-155 with a low detection limit down to 166.7 fM. In addition, the biosensor displayed a strong anti-interference ability in bovine whole blood, and thus was expected to be applied to detect blood samples of various miRNA-155 related diseases in clinical diagnosis.
研究领域:
生物化学
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分析化学
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生物医学工程
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分子生物学
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DOI
PubMed
高影响因子
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
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2022
·Vol. 414
·期 14
·4079-4088
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IF 4.478
Xu Wei,Zhang Yu,Chen Hao,Dong Jinhua,Khan Ranjha,Shen Jianjun,Liu Honglin
Here, we developed a surface-enhanced Raman scattering (SERS) sensor based on functionalized Au@Ag core–shell nanorods (Au@Ag NRs) and cascade DNAzyme amplifier (CSA) for sensitive and accurate determination of microRNA-21 (miRNA-21). The as-prepared SERS nanoprobes were composed of a thiol-modification hairpin probe (HP2)–functionalized Au@Ag NRs and hairpin DNAzyme (HP1-Dz). Compared with original gold nanorods, the silver shell caused an enhancement of plasmonic properties, resulting in a significant enhancement of Raman signals. In the presence of target miRNAs, the hairpin construction of HP1-Dz changed due to DNA/RNA hybridization; subsequently, the DNAzyme-catalyzed cleaving process changed, and the Raman signals of the SERS nanoprobes gradually “turned off” with time elapse because of the dissociation of the Raman reporter from the surface of Au@Ag NRs. Hence, based on this principle, the proposed SERS sensor exhibited good linearity in the range 0.5 fM to 10 nM for miRNA-21 detection with a detection limit (LOD) of 0.5 fM. The proposed SERS platform has potential application in quantitative and precise detection of miRNA-21 in human serum. Graphical abstract
研究领域:
纳米技术
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分析化学
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生物传感
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分子生物学
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生物医学工程
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DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2023
·
IF 8.4
Jiaxin Peng,Xiaomin Hang,Huiyi Wang,Kairen Zhao,Honghong Wang,Haoming Yu,Li Wang
MicroRNAs (miRNAs) have emerged as powerful biomarkers for various diseases diagnosis, yet their clinical application is greatly plagued by the lack of reliable and rapid detection methods. Here, a simple, fast, and accurate point-of-care testing (POCT) platform for miRNA-155 detection was developed by interfacing self-primer exponential amplification reaction (SPEXPAR) with colorimetric and POCT lateral flow assay (LFA). In this platform, a hairpin template was designed for exponential amplification reaction, which could effectively avoid nonspecific template interactions and eventually effectively avoid undesirable background amplification; the colorimetric assay is based on the SPEXPAR amplicons induced gold nanoparticles (AuNPs) color change; and the LFA detection principle involves the amplicons hybridization with designed probes. Under the optimized conditions, the colorimetric assay could detect miRNA-155 as low as 0.41 pM in less than 35 min; and LFA could detect as low as 100 pM in less than 40 min. Furthermore, the method was adopted to detect miRNA-155 in human serum samples with satisfactory recoveries. In summary, this study offered a promising analytical tool for molecular diagnostic testing application.
研究领域:
生物技术
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医学诊断
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分子生物学
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产品关联
DOI
高影响因子
Journal of Orthopaedic Surgery and Research
·
2022
·Vol. 17
·期 1
·1-10
·
IF 2.677
Bin-bin Zhang,Da-wa Zha Xi,Chao Li,Lan-tao Zhang,Tao Wu,Chuan Lu,Chao-zheng Liu,De-chun Li,Chang Feng,Shu-qing Wei,Zu-nan Dong,Xian-wei Pei,Zhang Zhi-xia,Ke-wen Li
Background Bone marrow mesenchymal stem cells have always been a heated research topic in bone tissue regeneration and repair because of their self-renewal and multi-differentiation potential. A large number of studies have been focused on finding the inducing factors that will promote the osteogenic differentiation of bone marrow mesenchymal stem cells. Previous studies have shown that macrophage exosomes or miRNA-26a-5p can make it work, but the function of this kind of substance on cell osteogenic differentiation has not been public.Methods M2 macrophages are obtained from IL-4 polarized bone marrow-derived macrophages. Exosomes were isolated from the supernatant of M2 macrophages and identified via transmission electron microscopy (TEM), western blotting, and DLS. Chondrogenic differentiation potential was detected by Alcian blue staining. Oil red O staining was used to detect the potential for lipogenic differentiation. And MTT would detect the proliferative capacity of cells. Western blot was performed to detect differential expression of osteogenic differentiation-related proteins.Results The results showed that M2 macrophage exosomes will promote bone differentiation and at the same time inhibit lipid differentiation. In addition, M2 macrophage-derived exosomes have the function of promoting the expression of SOX and Aggrecan suppressing the level of MMP13. The exosome inhibitor GW4689 suppresses miRNA-26a-5p in M2 macrophage exosomes, and the treated exosomes do not play an important role in promoting bone differentiation. Moreover, miRNA-26a-5p can enable to promote bone differentiation and inhibit lipid differentiation. miRNA-26a-5p can promote the expression of ALP (alkaline phosphatase), RUNX-2 (Runt-related transcription factor 2), OPN(osteopontin), and Col-2(collagen type II). Therefore, it is speculated that exosomal miRNA-26a-5p is indispensable in osteogenic differentiation.Conclusion sThe present study indicated that M2 macrophage exosomes carrying miRNA-26a-5p can induce osteogenic differentiation of bone marrow-derived stem cells to inhibit lipogenic differentiation, and miRNA-26a-5p will also promote the expression of osteogenic differentiation-related proteins ALP, RUNX-2, OPN, and Col-2.
研究领域:
干细胞生物学
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骨组织工程
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外泌体研究
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微小RNA功能
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DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2023
·
IF 7.4
Duo Chen,Na Chen,Fangning Liu,Yiming Wang,Huageng Liang,Yanbing Yang,Quan Yuan
Portable point-of-care testing (POCT) is currently drawing enormous attention owing to its great potential for disease diagnosis and personal health management. Electrochemical biosensors, with the intrinsic advantages of cost-effectiveness, fast response, ease of miniaturization, and integration, are considered as one of the most promising candidates for POCT application. However, the clinical application of electrochemical biosensors-based POCT is hindered by the decreased detection sensitivity due to the low abundance of disease-relevant biomolecules in extremely complex biological samples. Herein, we construct a flexible electrochemical biosensor based on single-stranded DNA functionalized single-walled carbon nanotubes (ssDNA-SWNTs) for high sensitivity and stability detection of miRNA-21 in human urine to achieve bladder cancer (BCa) diagnosis and classification. The ssDNA-SWNT electrodes with a 2D interconnected network structure exhibit a high electrical conductivity, thus enabling the ultrasensitive detection of miRNA-21 with a detection limit of 3.0 fM. Additionally, the intrinsic flexibility of ssDNA-SWNT electrodes endows the biosensors with the capability to achieve high stability detection of miRNA-21 even under large bending deformations. In a cohort of 40 BCa patients at stages I–III and 44 negative control samples, the constructed ssDNA-SWNT biosensors could detect BCa with a 92.5% sensitivity, an 88.6% specificity, and classify the cancer stages with an overall accuracy of 81.0%. Additionally, the flexible ssDNA-SWNT biosensors could also be utilized for treatment efficiency assessment and cancer recurrence monitoring. Owing to their excellent sensitivity and stability, the designed flexible ssDNA-SWNT biosensors in this work propose a strategy to realize point-of-care detection of complex clinical samples to achieve personalized healthcare.
研究领域:
生物医学工程
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纳米技术
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癌症诊断
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电化学
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即时检测设备
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DOI
PubMed
高影响因子
NEW JOURNAL OF CHEMISTRY
·
2021
·Vol. 45
·期 24
·10907-10913
·
IF 3.591
Yanfang Cui,Jing Zheng,Wei Zhuang,Haiwang Wang
A novel surface-enhanced Raman scattering (SERS) platform has been developed for the highly sensitive detection of miRNA-21 by target activation depending on exonuclease-assisted signal amplification with a plasmon coupling phenomenon. In this system, a Raman dye-modified gold nanoparticle (AuNP) substrate can be covalently linked by the thiolated DNA track and DNA walker, which combine the protection probe at the 3′ terminal end to prevent reaction with the DNA track. In the presence of miRNA-21, it will combine the protect probe to the DNA walker depending on more complementary base pairing and release the DNA walker, which will combine with the adjacent DNA track. Depending on the exonuclease III (exo III) cleavage, the DNA walker can walk along the surface of AuNPs and produce the self-assembled hot spots between AuNPs, allowing a homogenous detection of the miRNA-21 with high sensitivity and reproducibility. Under optimized experimental conditions, the platform detected miRNA-21 with a wide range from 10 pM to 1.0 × 105 pM and displayed very low detection limit (4.0 pM). Hence, the novel strategy will become a promising tool for the SERS assay of miRNA-21 with high sensitivity and repeatability. Furthermore, this platform also owns great potential for cancer typing and clinical diagnostics.
研究领域:
分析化学
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纳米技术
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生物医学工程
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分子生物学
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癌症研究
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产品关联
DOI
高影响因子
ACS Applied Materials & Interfaces
·
2022
·
IF 10.383
Jiulong Zhang,Xiaoyan Sun,Xiufeng Zhao,Lin Liu,Xin Cheng,Chunrong Yang,Haiyang Hu,Mingxi Qiao,Dawei Chen,Xiuli Zhao
The tumor immunosuppressive microenvironment (TIM) greatly hindered the efficacy of cancer immunotherapy. Overexpressed indoleamine 2,3-dioxygenase-1 (IDO1) in tumor tissues plays a vital role in TIM generation, and downregulation of IDO1 expression may reverse TIM. Inspired by the Watson–Crick base-pairing rule, a versatile noncationic miRNA vector (miDAC@PDA) is developed for cancer immunotherapy. Doxorubicin (DOX), adenosine triphosphate (ATP), and copper ions (Cu2+) are coassembled into coordination polymer nanoparticles (DAC) and bind miRNA via the hydrogen bond interaction (miDAC) between adenine residues (ATP) and uracil residues (miRNA). Polydopamine (PDA) is deposited onto the surface of miDAC for photothermal therapy. miDAC@PDA can efficiently accumulate into tumor tissues for cellular uptake. Under laser irradiation and high intracellular GSH levels, the PDA shell of miDAC@PDA can dissociate from miDAC for miRNA release due to local hyperthermia. Cu2+-mediated GSH consumption and intracellular ATP release can amplify the DOX-based immunogenic cell death (ICD) cascade, together with miR-448-mediated IDO1 inhibition, and these versatile nanoplexes will not only restrain primary tumor growth but also display a remarkable abscopal effect on distant tumors. Collectively, our study provides a unique strategy for intracellular gene delivery and an inspirational approach for multimechanism cancer management.
研究领域:
癌症研究
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纳米技术
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治疗学
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生物医学工程
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分子生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Oxidative Medicine and Cellular Longevity
·
2019
·
IF 4.868
Lifang Sun,Min Zhu,Wei Feng,Yiping Lin,Jia Yin,Juan Jin,Yunguang Wang
Idiopathic pulmonary fibrosis (IPF) is a prototype of chronic, progressive, and fibrotic lung disease with high morbidity and high mortality. Menstrual blood-derived stem cells (MenSCs) have proven to be an attractive tool for the treatment of acute lung injury and fibrosis-related diseases through immunosuppression and antifibrosis. However, whether MenSC-derived exosomes have the similar function on pulmonary fibrosis remains unclear. In the present study, exosomes secreted from MenSCs (MenSCs-Exo) were verified by transmission electron microscope (TEM), nanoparticle tracking analyzer (NTA), and western blotting. And MenSC-Exo addition significantly improved BLM-induced lung fibrosis and alveolar epithelial cell damage in mice, mainly reflected in BLM-mediated enhancement of the fibrosis score, blue collagen deposition, dry/wet gravity ratio, hydroxyproline and malondialdehyde levels, and downregulation of glutathione peroxidase, which were all robustly reversed by MenSC-Exo management. Additionally, BLM- and TGF-β1-evoked cellular reactive oxygen species (ROS), mitochondrial DNA (mtDNA) damage, and cell apoptosis were rescued by MenSCs-Exo in vivo and in vitro. Further study indicated that the MenSCs-Exo could transport miRNA Let-7 into recipient alveolar epithelial cells. Let-7 inhibitor administration significantly blocked the exosome-mediated improvement role on lung fibrosis in mice. Mechanistically, Let-7 was able to regulate the expression of lectin-like oxidized low-density lipoprotein receptor-1 (LOX1) through binding to its 3-UTR region. Forced expression of LOX1 promoted the expression of apoptosis-related protein and mtDNA damage markers via regulating NLRP3 which was also confirmed in BLM model mice under the combination therapy of the exosome and Let-7 inhibitor. Collectively, this study demonstrates that exosomal Let-7 from MenSCs remits pulmonary fibrosis through regulating ROS, mtDNA damage, and NLRP3 inflammasome activation. This provides a new approach of exocytosis on the treatment of fibrotic lung disease.
研究领域:
干细胞生物学
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肺医学
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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
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