Nanoscale
·
2025
·
IF 5.1
Beibei Zhang,Xinyi Wang,Min Pan,Xinhao Li,Hong Wang,Yingwei Zhang
In this study, a novel dual-switch fluorescent biosensor based on a bipedal DNAzyme walker was developed for the highly sensitive detection of the tumour biomarker miRNA-141. By exploiting the typically elevated expression levels of miRNA-141 and apurinic/apyrimidinic endonuclease 1 (APE1) in tumour cells, an endogenous dual-gating system was constructed. The presence of both targets was required to initiate an enzyme-free strand displacement reaction, enabling target recycling and the subsequent release of bipedal DNAzyme walker components (D1/D2), which were efficiently isolated using streptavidin-modified magnetic beads. In the presence of Mg2+, the activated DNAzyme walker specifically cleaved fluorophore-labelled substrate strands on gold nanoparticles (AuNPs). Through repeated“walking–cleavage”cycles, fluorescence signals were progressively restored and amplified. By integrating AuNPs as carriers and magnetic separation technology, a cascade dual-amplification system based on non-enzymatic strand displacement and bipedal DNAzyme walking was established, markedly enhancing the detection sensitivity. The biosensor exhibited dual-switch-specific activation, effectively suppressing interference from non-target RNAs and other nucleases, and maintained a low background signal. It demonstrated excellent discrimination between tumour cells and normal cells, and showed good anti-interference performance in spiked serum samples. This study provides a new strategy for the development of high-precision tumour biomarker detection, offering potential for trace analysis in complex biological samples.
研究领域:
生物传感器
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分子诊断
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核酸化学
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肿瘤标志物
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纳米技术
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DOI
PubMed
高影响因子
MICROCHEMICAL JOURNAL
·
2025
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IF 5.1
Weiying Liu,Xiaolu Sui,Yunpeng Xu,Tingfei Xie,Yanzi Zhang,Aisha Zhang,Zhenyu Lin,Jihong Chen
MicroRNA-21 (miRNA-21) can serve as an early biomarker for acute kidney injury (AKI), it is important to develop sensitive and selective detection methods for miRNA-21 detection. In this study, an electrochemiluminescence (ECL) biosensor for miRNA-21which synergistically integrates T7 RNA polymerase-driven transcriptional amplification with CRISPR/Cas13a-mediated collateral cleavage and magnetic separation strategy had been designed. T7 RNA polymerase-driven transcriptional amplification has the characters of low requirements for conditions and high amplification efficiency. A cascaded signal amplification strategy which coupled enzymatic target recycling with CRISPR/Cas13a-mediated exponential signal amplification was proposed to achieve high detection sensitivity. And T7 promoter specificity and CRISPR/Cas13a sequence targeting had been adopted to ensure high specificity of the developed system. Furthermore, magnetic bead separation had been adopted to reduce the background signal. The parameters which may affected the performance of the biosensor had been optimized. Under optimized conditions, the ECL response of the system has a linear relationship with the logarithmic value of miRNA-21 concentration in the range of 1.0 fM to 1.0 nM, with a calculated limit of detection(LOD) of 0.33 fM (S/N = 3). The proposed methodology has been adopted to detect the target in clinical urine specimens with satisfied results.
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
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生物医学工程
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临床诊断
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DOI
高影响因子
Journal of Materials Chemistry B
·
2021
·Vol. 9
·期 15
·3335-3345
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IF 6.331
Chen Tan,Fan Jia,Peng Zhang,Xinghuai Sun,Yunsheng Qiao,Xueli Chen,Youxiang Wang,Junyi Chen,Yuan Lei
The elevation of intraocular pressure (IOP) is an important risk factor in the development of primary open angle glaucoma (POAG), which is the main cause of irreversible vision loss. miRNAs are promising new anti-glaucoma therapeutic agents. However, the low stability and cellular transfection of miRNA in vivo hinder its further application. This study aims to investigate the use of polydopamine–polyethylenimine nanoparticles (PDA/PEI NPs) as miRNA carriers in the treatment of ocular hypertension and glaucoma. The in vitro study proves that the carrier preserves the activity of nucleic acid for a long period. Besides, it has comparable transfection efficiency with commercially available vehicles, while having lower cytotoxicity. It has been demonstrated in the animal model that PDA/PEI NPs successfully reach the target tissues without an obvious inflammatory response. PDA/PEI NPs/miR-21-5p increases the permeability of porcine angular aqueous plexus cells, thereby reducing IOP by facilitating the conventional outflow pathway at least partially through the pathway involving endothelial nitric oxide synthase. Our results indicate that PDA/PEI NPs/miR-21-5p is a promising anti-glaucoma drug for treating POAG. And the delivery strategy may be extended to other gene therapy in treating intraocular diseases.
研究领域:
药学
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纳米技术
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眼科
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分子生物学
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DOI
PubMed
高影响因子
JOURNAL OF APPLIED TOXICOLOGY
·
2023
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IF 3.3
Tianqi Huang,Jinglou Chen,Yumiao Zhang,Yao Chen,Congyue Xu,Jing Guo,Hao Ming
Cadmium (Cd) is a toxic heavy metal pollutant and serves as an important environmental endocrine-disrupting chemical. Cd exposure is believed to can enhance the risks of age-related disorders including benign prostatic hyperplasia (BPH). This study was to investigate the harms of Cd exposure on mice prostate and human nonmalignant prostate epithelial RWPE-1 cells. Mice prostate fibrosis was evaluated by visualizing the prostatic collagen deposition via Masson and Sirius red staining, and detecting the content of hydroxyproline. Additionally, the epithelial-mesenchymal transition (EMT), primary ciliogenesis and SHH signaling pathways in both mice prostate and RWPE-1 cells were evaluated. It was found that Cd exposure stimulated prostatic collagen deposition, EMT and primary ciliogenesis, as well as enhanced the circ_0027470 level and reduced the miRNA-1236-3p level. Circ_0027470 functioned as a sponge of miRNA-1236-3p, which had the inhibiting target of SHH. The whole results showed that circ_0027470 promoted Cd exposure-induced prostatic fibrosis via sponging miRNA-1236-3p and subsequently stimulating SHH signaling pathway. This study shed a light on a novel molecular mechanism involved in circRNA for Cd exposure-induced prostate deficits.
研究领域:
毒理学
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分子生物学
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细胞生物学
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环境健康
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DOI
PubMed
高影响因子
Scientific Reports
·
2017
·Vol. 7
·期 1
·1-12
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IF 4.259
Liu Liang,Xu Qingqing,Hao Shuai,Chen Yun
MicroRNAs (miRNAs) play a vital role in regulating gene expression and are associated with a variety of cancers, including breast cancer. Their distorted and unique expression is a potential marker in clinical diagnoses and prognoses. Thus, accurate determination of miRNA expression levels is a prerequisite for their applications. However, the assays currently available for miRNA detection typically require pre-enrichment, amplification and labeling steps, and most of the assays are only semi-quantitative. Therefore, we developed a quasi-direct liquid chromatography-tandem mass spectrometry (LC-MS/MS)-based targeted proteomics approach to quantify target miRNA by innovatively converting the miRNA signal into the mass response of a reporter peptide via a covalently immobilized DNA-peptide probe. Specifically, the probe containing the targeted proteomics-selected substrate/reporter peptide, GDR AVQLGVDPFR/AVQLGVDPFR, and the DNA sequence complementary to the target miRNA (i.e., miR-21) was first immobilized on APMTS modified silica nanoparticles using PDITC. After the immobilized probe was recognized and hybridized with the target miRNA, the excess probe was degraded using MBN and followed by a trypsin digestion of the hybrids. The reporter peptide was released and quantified using LC-MS/MS. The obtained LOQ was 5 pM. Finally, the developed assay was used for the quantitative analysis of miR-21 in breast cells and tissue samples.
研究领域:
分子生物学
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癌症研究
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蛋白质组学
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分析化学
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DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
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2021
·
IF 7.46
Lei Liu,Yue Yao,Kejian Ma,Changjian Shangguan,Songlong Jiao,Songyang Zhu,Xiaoxuan Xu
Elucidation of the photoelectrochemical (PEC) detection mechanism and exploring highly efficient photoactive materials hold both a challenge and an opportunity to construct high-performance biosensors. In this work, we present a strategy to enhance the charge separation efficiency by fabricating hierarchical Ti 3 C 2 @ReS 2 via the vertical anchoring flaky ReS 2 on the Ti 3 C 2 backbone. According to the experimental results, the Ti 3 C 2 @ReS 2 sample containing 45 wt % of ReS 2 exhibits a 2.48-time promotion in the photocurrent as compared to ReS 2 owing to the synergistic effects of its photoactive and conductive counterparts, indicating its significant advantages in separating photo-induced carriers and suppressing carrier recombination. In essence, the Ti 3 C 2 @ReS 2 Schottky junction formed at the interface can capture the photogenerated electrons generated by ReS 2 , thereby suppressing carrier recombination. As a result, an ultrasensitive PEC detection platform based on the optimized Ti 3 C 2 @ReS 2 can offer a log-linear detection window from 0.1 fM to 1 nM with an estimated detection limit of 2.4 aM (S/N = 3) for cancer-related miRNA-141 without any other amplification.
研究领域:
材料科学
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生物传感器
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癌症研究
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光电化学
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DOI
高影响因子
CELL BIOLOGY INTERNATIONAL
·
2022
·Vol. 46
·期 12
·2220-2231
·
IF 4.473
Maomao Miao,Yang Zhang,Xuping Wang,Shanshan Lei,Xiaowen Huang,Luping Qin,Dan Shou
Diabetic osteoporosis (DOP) is a disorder of bone metabolism induced by multiple mechanisms. Previous studies have revealed that microRNAs (miRNAs) play crucial roles in bone metabolism. MiRNA-144-5p has been proven to participate in the regulation of osteoblast activities; however, its specific mechanism in DOP has not been elucidated. This study investigated whether high glucose (HG) inhibited osteoblasts by regulating miRNA-144-5p. Our results showed that HG inhibited bone formation not only in vivo but also in vitro. We observed that HG severely hindered the migration, proliferation and mineralization of osteoblasts, while miRNA-144-5p was upregulated by way of the cell counting kit-8 assay, wound healing assay, alkaline phosphatase (ALP) activity assay and alizarin red staining. Double luciferase reporter experiments showed that miRNA-144-5p directly targeted insulin receptor substrate 1 (IRS1). The IRS1/AKT signaling pathway is closely related to osteoblasts' migration, proliferation, and mineralization. Silencing miRNA-144-5p promoted the mRNA, and protein expression of IRS1, thereby letting the expression of total AKT down, and then preventing phosphorylation of AKT into the nucleus to regulate migration, proliferation, and mineralization genes of osteoblasts. In conclusion, this study indicated that HG regulated the migration, proliferation, and mineralization of osteoblasts via the miRNA-144-5p/IRS1/AKT axis, which suggested a possible mechanism for DOP pathology.
研究领域:
分子生物学
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内分泌学
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骨代谢
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糖尿病
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DOI
PubMed
高影响因子
Discovery Medicine
·
2024
·Vol. 36
·期 186
·1420-1429
·
IF 2
Shimaa Saad El-Din,Basma Emad Aboulhoda,Amira Hassouna,Marwa M Shakweer,Mansour A Alghamdi,Doha Essam,Mohamed Essam,Nawaf A AlRakaf,Hanaa M Elzahed,Asmaa Selmy,Dina Sabry,Dina M Mekawy
Background: Bone marrow-derived mesenchymal stem cells (BM-MSCs) have recently attracted great attention due to their crucial anti-inflammatory and regenerative properties. This work aims to examine the curative impact of intra-articular injection of BM-MSCs-derived exosomes in ameliorating osteoarthritis (OA) progression in rats and to explore the interaction between circular RNA of Yes-associated protein 1 ( circYAP1 ) and microRNA-21 ( miRNA-21 ) in the rat knee joints. Methodology: Gene expression circYAP1 , miRNA-21 , toll-like receptor-7 ( TLR7 ), aggrecan, and collagen type II were evaluated by quantitative reverse transcription polymerase chain reaction (qRT-PCR) in the rat articular tissues. In addition, the Enzyme-linked immunosorbent assay (ELISA) technique was used to estimate the level of the inflammatory markers interleukin 4 (IL-4), interleukin 10 (IL-10), interleukin 1β (IL-1β), and tumor necrosis factor-alpha (TNF-α); and the oxidative markers glutathione (GSH), malondialdehyde (MDA) and total reactive oxygen species (ROS). Histopathological examination using Hematoxylin and Eosin (H&E) staining of the rat articular tissue was also performed along with an estimation of the articular cartilage thickness. Results: Our results showed that BM-MSCs-derived exosomes significantly elevated circYAP1 gene expression level ( p < 0.05) along with subsequent downregulation of miRNA-21 and TLR7 ( p < 0.05). These effects impacted the inflammatory milieu of rat articular surfaces, where there was a significant reduction ( p < 0.05) of the pro-inflammatory and oxidative markers with significantly increased production of the anti-inflammatory and antioxidative markers ( p < 0.05). Marked elevation in aggrecan and collagen type II gene expression was also found in the treated groups ( p < 0.05). Conclusion: Those data suggest that BM-MSCs-derived exosomes have a crucial role in mitigating OA symptoms and pathology progression and might be regarded as an effective as well as acceptable treatment option for OA.
研究领域:
再生医学
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干细胞生物学
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分子生物学
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骨科
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DOI
PubMed
高影响因子
JOURNAL OF ETHNOPHARMACOLOGY
·
2023
·
IF 5.4
Ye Liu,Jiufeng Zhao,Xingrui Mu,Junyu Deng,Xingqian Wu,Wenjie He,Yiqiu Liu,Rifang Gu,Felicity Han,Xuqiang Nie
Ethnopharmacological relevance Centella asiatica (L.) Urban is an ethnobotanical herb. The main bioactive components of Centella asiatica are pentacyclic triterpenoid glycosides , namely asiaticoside and hydroxyasiaticoside. Asiaticoside possess a diverse array of pharmacological properties, such as wound-healing, anti-inflammatory, antioxidant, anti-allergic, antidepressant, anxiolytic, anti-fibrotic, antibacterial, anti-arthritic, anti-tumor, and immunomodulatory activities. Aim of the study The purpose of this investigation is to explore potential therapeutic interventions for the delayed healing of wounds in diabetic patients (DW) facilitated by Asiaticoside-Nitric Oxide. To clarify the key molecular mechanism of miRNA-21-5p in DW wound repair and to deepen the understanding of DW disease pathogenesis. Materials and methods Firstly, miRNA microarray technology, bioinformatics, and RT-qPCR were used to analyze DW patients' and normal controls' skin tissue samples. Secondly, in order to investigate the role of miRNA-21-5p, a hyperglycemic model was established using HaCaT cells. Overexpressing as well as interfering HaCaT cell lines were constructed by lentiviral infection to further explore the proliferative and migratory effects of Asiaticoside-Nitric Oxide. The next step was to search for potential target genes of miRNA-21-5p and verify them with dual-luciferase reporter assay. Finally, the expression levels of target genes and proteins were detected through the utilization of RT-qPCR and Western blotting under the influence of Asiaticoside-Nitric Oxide. Results A library of miRNAs and target genes expressed explicitly in DW patients and rats was established. The study confirmed the upregulation of miRNA-21-5p in DW patients and identified its involvement in signaling pathways related to chronic ulcer wound repair. Overexpression of LV-miRNA-21-5p significantly promoted cell proliferation , while treatments of Asiaticoside-Low dose (AC-L) and Asiaticoside-Medium dose (AC-M) enhanced proliferation and migration, particularly when combined with nitroprusside (SNP). Further analysis revealed potential target genes of miRNA-21-5p, such as TGF-β1, SMAD7, and TIMP3. Their interaction with miRNA-21-5p was confirmed through dual luciferase assays. The study found that anti-DW drugs increased the expression of TGF-β1 and SMAD7 while inhibiting TIMP3 expression in a high-glucose environment. Conclusions The research concluded that miRNA-21-5p plays a crucial role in the delayed healing of diabetic wounds, and that the combination treatment of AC + SNP shows promise in promoting wound healing in DW rats. Target genes, including TGF-β1, SMAD7, and TIMP3, may contribute to the regulatory mechanisms involved in diabetic wound healing. These findings provide valuable insights for developing novel therapeutic approaches for DW.
研究领域:
药理学
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分子生物学
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糖尿病研究
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伤口愈合
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民族药理学
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产品关联
DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2023
·
IF 8.4
Jiajing Xu,Weiqiang Yang,Jiancong Ni,Qingxiang Wang,Zhenyu Lin
Electrochemiluminescence (ECL) arises from the collisional annihilation between the electro-excited luminophore and co-reactant. Most ECL-based bioassays require extensive free co-reactants that collide with luminophores randomly. Herein, the N,N-dimethylaniline (DMA) group was electro-grafted on the ITO electrode surface as the bottom co-reactant layer, and the vertically ordered mesoporous silica film (VMSF) was prepared as the upper layer to regulate the accessibility of Ru(bpy) 3 2+ . Thus, the dual-layer functionalized VMSF/N-ITO was newly built for the controllable ECL study. Relevant conditions, including the DMA electro-grafting, VMSF thickness, buffer concentration and pH value, were discussed to improve the ECL efficiency. Based on the VMSF/N-ITO platform, a homogeneous ECL bioassay was developed for miRNA-21 detection. The target nucleotide sequence induced the hybridization chain reaction (HCR) of probes H1 and H2 to absorb free Ru(bpy) 3 2+ as much as possible. Residual Ru(bpy) 3 2+ permeated the VMSF nanochannels to collide with the DMA co-reactant in producing an ECL response. There was a linear relationship between ∆ECL intensity and logarithmic concentration of miRNA-21 in the 1 fM ∼ 100 pM range, and the detection limit was 0.25 fM. The developed approach was probes immobilization-free, highly sensitive, and anti-interference, opening a new avenue for ECL-based bioassays.
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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生物传感器
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DOI
高影响因子
Chemical Science
·
2016
·Vol. 7
·期 7
·4184-4189
·
IF 9.144
Zhen Zhang,Yuanyuan Wang,Ningbo Zhang,Shusheng Zhang
We herein report a novel finding that nucleic acid molecular aggregates (NAMAs) self-assembled on graphene oxide nanoplates (GONPs) as a result of DNA rolling circle amplification (RCA) and a functionalized triple-helix probe (THP) in single cells. The functionalized THP containing the aptamer region for target recognition and the trigger DNA region for RCA was firstly used to activate RCA for miRNA imaging in single cells. Interestingly, NAMAs with the fluorescent labels were hybridized by both the RCA products and FAM-DNA, and could partly self-assemble on GONPs; meanwhile, NAMAs could extend from the GONPs, which led to the quenched fluorescence being renewed. Significantly, the NAMAs were successfully applied for low-abundance miRNA detection and imaging in single cells. The self-assembled NAMAs could generate prominent and agminated fluorescence-bright spots in single cancer cells, which will effectively drive cell imaging into a new era.
研究领域:
分子生物学
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纳米技术
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生物医学工程
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细胞生物学
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产品关联
DOI
PubMed
高影响因子
International Journal of Nanomedicine
·
2024
·
IF 6.6
Chen Lin,Xinyi Bai,Linkun Zhang
Purpose MicroRNA-34c-5p (miR-34c-5p) plays a pivotal role in bone remodeling, yet its therapeutic potential is hindered by challenges such as instability, limited cellular internalization, and immune responses. This study was aimed at developing innovative scaffolds capable of efficiently delivering microRNAs (miRNAs), specifically miR-34c-5p.Methods Chitosan (CS)/sodium tripolyphosphate (STPP)/sodium alginate (SA) scaffolds, referred to as CTS scaffolds, were synthesized at a specific ratio and characterized using dynamic light scattering and scanning electron microscopy (SEM). Cytotoxicity assessments were conducted through cell activity staining. The loading capacity and releasing performance of miRNAs were quantified using spectrophotometry. Subsequently, the in vivo efficacy of miR-34c-5p Agomir/Antagomir in regulating bone repair was evaluated in the rabbit cranial bone defect model, with micro-CT scanning and histological analysis conducted at 4, 8, and 12 weeks.Results CTS scaffolds with a composition ratio of 1:0.2:0.1 were successfully synthesized, exhibiting a mean particle size of 360.1 nm. SEM revealed scaffolds had the porous spongy structure. Cell activity staining confirmed the excellent biocompatibility of the CTS scaffolds. Spectrophotometry demonstrated miR-34c-5p Antagomir were continually released, reaching 91.41% within 30 days. Differential new bone formation was observed between the miR-34c-5p Agomir and Antagomir groups. Micro-CT imaging and histological staining revealed varying degrees of bone regeneration, with notable improvements in the miR-34c-5p Antagomir group.Conclusion CTS scaffolds with a composition ratio of 1:0.2:0.1 demonstrate favorable biocompatibility and enable efficient loading and sustained release of miR-34c-5p Antagomir. The study suggests potential applications of miR-34c-5p Antagomir in promoting bone repair and highlights the promise of innovative scaffolds for therapeutic miRNAs administration in bone regeneration.
研究领域:
生物医学工程
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组织工程
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再生医学
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分子生物学
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DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）