TALANTA
·
2021
·
IF 6.057
Min-Xi Li,Yao Chen,Zeng-Ping Chen,Ru-Qin Yu
Herein, a label free and sensitive miRNA detection method with enhanced practical applicability was developed based on the locked nucleic acid (LNA) assisted repeated fishing amplification strategy. The working mechanism of the proposed method is as follows: 1) a DNA probe (i.e, L-DNA) with LNA bases is immobilized onto the surface of a gold foil. The L-DNA hybridizes with the 3′ terminus of the first strands of complementary deoxyribonucleic acid (cDNA) of the target miRNA in the test samples; 2) The protruding 5′ terminus of the cDNA serves as a ‘fishhook’ to repeatedly fish the products of a hybridization chain reaction (HCR) out from a ‘reaction tube’; 3) The HCR products can be unloaded from the gold foil into a ‘product tube’ through temperature-controlled dehybridization; 4) The concentration of the target miRNA is determined based on the fluorescence intensity generated by the addition of SYBR-Green I (SG) into the ‘product tube’. The proposed platform was applied to the detection of miRNA-122 in cell lysate samples and obtained quantitative results with accuracy comparable to the quantitative reverse transcription PCR method (qRT-PCR). It is worth pointing out that the proposed platform achieved a limit of detection value of 2.9 fM for miRNA-122 by a simple but effective LNA-assisted repeated fishing amplification strategy instead of complicated enzyme-based amplification techniques. It is reasonable to expect that the proposed method provides a competitive alternative for designing practically applicable, cost-effective and label-free miRNA detection methods.
研究领域:
生物化学
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分子生物学
·
纳米技术
·
分析化学
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产品关联
DOI
PubMed
高影响因子
POSTHARVEST BIOLOGY AND TECHNOLOGY
·
2023
·
IF 7
Fan Yang,Shuqi Liu,Zhonghuan Tian,Yujie Du,Deyao Zhang,Chao-an Long
Edetate salts are a kind of food additive regarded as candidates for chemical fungicides because of their lower toxicity and higher safety for the environment and human health. The antifungal activities of edetate salts have been determined in some phytopathogens except Penicillium digitatum , which causes the most destructive fungal disease during the citrus postharvest period. In the present study, we investigated the molecular mechanisms of calcium disodium edetate (EDTA-Na 2 Ca) against P. digitatum in deepth. The results suggested that EDTA-Na 2 Ca inhibited the P. digitatum P44 strain in vitro at a minimum inhibitory/fungicidal concentration (MIC/MFC) of 50 mmol L −1 . Meanwhile, EDTA-Na 2 Ca also inhibited P. digitatum in vivo by enhancing the host defense system. The P44 strain treated with EDTA-Na2Ca displayed a delayed conidial germination rate, expanded hyphal tips, thickened septa and cell walls, and destroyed mitochondria. Transcriptome sequencing and qRT-PCR analyses indicated that EDTA-Na 2 Ca suppressed the P44 strain primarily by disturbing the carbohydrate metabolism pathway, especially the synthesis of pyruvate in glycolysis. In addition, the adverse effects, including those on mycelial growth, pyruvate synthesis and superoxide dismutase activity, of EDTA-Na 2 Ca could be reversed by exogenous Mn 2+ , implying that Mn 2+ chelation is an essential mode of action. In summary, the current work verifies that EDTA-Na 2 Ca is a potential antifungal drug that cures citrus green mold probably by chelating Mn 2+ and targeting pyruvate synthesis in P. digitatum .
研究领域:
植物病理学
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分子生物学
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食品科学
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环境科学
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DOI
高影响因子
PHYTOMEDICINE
·
2021
·
IF 5.34
Ying-yuan Zhao,Xin-peng Yao,Ting-ying Jiao,Jia-ning Tian,Yue Gao,Shi-cheng Fan,Pan-pan Chen,Yi-ming Jiang,Yan-ying Zhou,Yi-xin Chen,Xiao Yang,Min Huang,Hui-chang Bi
Background Schisandrol B (SolB) is one of the bioactive components from a traditional Chinese medicine Schisandra chinensis or Schisandra sphenanthera . It has been demonstrated that SolB exerts hepatoprotective effects against drug-induced liver injury and promotes liver regeneration . It was found that SolB can induce hepatomegaly but the involved mechanisms remain unknown. Purpose This study aimed to explore the mechanisms involved in SolB-induced hepatomegaly. Methods Male C57BL/6 mice were injected intraperitoneally with SolB (100 mg/kg) for 5 days. Serum and liver samples were collected for biochemical and histological analyses. The mechanisms of SolB were investigated by qRT-PCR and western blot analyses, luciferase reporter gene assays and immunofluorescence. Results SolB significantly increased hepatocyte size and proliferation, and then promoted liver enlargement without liver injury and inflammation. SolB transactivated human PXR , activated PXR in mice and upregulated hepatic expression of its downstream proteins, such as CYP3A11 , CYP2B10 and UGT1A1. SolB also significantly enhanced nuclear translocation of PXR and YAP in human cell lines. YAP signal pathway was activated by SolB in mice. Conclusion These findings demonstrated that SolB can significantly induce liver enlargement, which is associated with the activation of PXR and YAP pathways.
研究领域:
药理学
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分子生物学
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肝病学
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产品关联
DOI
PubMed
高影响因子
ACTA BIOCHIMICA ET BIOPHYSICA SINICA
·
2020
·Vol. 52
·期 8
·842-852
·
IF 2.836
Zhang Liang,Wang Xin,Liu Xin,Lv Mingyue,Shen Erdong,Zhu Guolian,Sun Zhe
Esophageal cancer (EC) is one of the most common aggressive malignant diseases worldwide. miR-28-5p plays important regulatory roles in many cancers including human EC. However, the molecular mechanism and potential role of miR-28-5p in EC remain uncertain. In this study, qRT-PCR and western blot analysis revealed that miR-28-5p expression was up-regulated and metastasis suppressor-1 (MTSS1) was down-regulated in EC tissues relative to matched para-cancer tissues. Cell counting kit-8 (CCK-8) assay demonstrated that miR-28-5p mimics increased cell viability, and miR-28-5p inhibitor decreased it. Flow cytometry (FCM) assay indicated that miR-28-5p mimics promoted cell cycle entry, while miR-28-5p inhibitor reduced it and induced cell apoptosis. Moreover, miR-28-5p mimics up-regulated the expressions of cyclin A, cyclin dependent kinase 2 (CDK2), cyclin D1, and cyclin E but down-regulated the expressions of cleaved caspase-3 and cleaved caspase-9, which was abolished by miR-28-5p inhibitor. Furthermore, luciferase reporter assay verified that miR-28-5p directly targeted MTSS1 3′UTR and down-regulated its expression. MTSS1 overexpression in TE-1 cells inhibited cell proliferation and promoted apoptosis induced by miR-28-5p mimics, whereas silencing of MTSS1 reversed cell progression induced by miR-28-5p inhibitor. We also demonstrated that miR-28-5p could promote esophageal tumor formation in vivo. Hematoxylin–eosin staining, immunohistochemistry, and TUNEL assays confirmed that miR-28-5p antagomir inhibited cell growth and accelerated apoptosis. Our results suggest that miR-28-5p may induce cell proliferation and suppress apoptosis to promote EC tumor formation via decreasing MTSS1 expression. Thus, miR-28-5p may be a potential target for human EC therapy.
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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产品关联
DOI
PubMed
高影响因子
ARCHIVES OF ORAL BIOLOGY
·
2020
·
IF 1.931
Ying Bian,Juan Xiang
Background Osteogenic differentiation of human periodontal ligament cells (hPDLCs) is crucial for regenerate periodontal tissues. In this study, we investigated the function of salvianolic acid B (Sal B) in osteogenesis of hPDLCs. Methods HPDLCs were isolated from healthy third molar roots. HPDLCs at passage 3 were identified by morphological observation and immunohistochemistry of vimentin . The viability of hPDLCs incubated with Sal B at concentrations of 0μM, 0.1μM, 0.5μM, 1μM and 5μM were measured by CCK-8 assay. To evaluate the effect of Sal B on osteogenic differentiation of hPDLCs, the alkaline phosphatase (ALP) activity, osteogenic differentiation markers, and mineralized nodules were determined by ALP kit, qRT-PCR and alizarin red S staining, respectively. To confirm the function of Sal B in hPDLCs involved in Wnt/β-catenin signaling pathway , hPDLCs were incubated with Sal B or co-incubated with Sal B and DKK-1 (a inhibitor of Wnt/β-catenin). The levels of Wnt/β-catenin signaling pathway and osteogenic differentiation-associated indicators were then determined. Results HPDLCs showed a typical fibroblast-like and spindle-shaped, with vimentin-positive. The viability of hPDLCs had no obvious change with stimulation of Sal B at various doses. Sal B promoted the increase of ALP activity, osteogenic differentiation markers levels, mineralized nodules and activation of Wnt/β-catenin signaling pathway, and DKK-1 could block those effects of Sal B on hPDLCs. Conclusion Sal B promoted osteogenesis of hPDLCs through Wnt/β-catenin signaling pathway, which providing a potential drug for periodontitis treatment .
研究领域:
牙科学
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药理学
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细胞生物学
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分子生物学
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产品关联
DOI
PubMed
高影响因子
Theranostics
·
2017
·
IF 8.766
Feifei Xu,Weixian Zhou,Jianxiang Cao,Qingqing Xu,Dechen Jiang,Yun Chen
The distorted and unique expression of microRNAs (miRNAs) in cancer makes them an attractive source of biomarker. There is much evidence indicating that a panel of miRNAs, termed “miRNA fingerprints”, is more specific and informative than an individual miRNA as biomarker. Thus, multiplex assays for simultaneous quantification of multiple miRNAs could be more potent in clinical practice. However, current available assays normally require pre-enrichment, amplification and labeling steps, and most of them are semi-quantitative or lack of multiplexing capability. In this study, we developed a quasi-targeted proteomics assay for multiplexed miRNA quantification by a combination of DNA-peptide probes and liquid chromatography-tandem mass spectrometry (LC-MS/MS). Specifically, the signal of target miRNAs (i.e., miR-21, miR-let7a, miR-200c, miR-125a and miR-15b) was converted into the mass response of reporter peptides by hybridization of miRNAs with DNA-peptide probes and subsequent tryptic digestion to release the peptides. After a careful optimization of conditions related to binding, conjugation, hybridization and multiple reaction monitoring (MRM) detection, the assay was validated for each miRNA and the limit of quantification (LOQ) for all the miRNAs can achieve 1 pM. Moreover, crosstalk between DNA-peptide probes in multiplex assay was sophisticatedly evaluated. Using this quasi-targeted proteomics assay, the level of target miRNAs was determined in 3 human breast cell lines and 36 matched pairs of breast tissue samples. Finally, simplex assay and qRT-PCR were also performed for a comparison. This approach grafts the strategy of targeted proteomics into miRNA quantification and may offer a new way for multiplexed miRNA profiling.
研究领域:
癌症生物学
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蛋白质组学
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分子生物学
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生物化学
·
临床诊断
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产品关联
DOI
PubMed
高影响因子
AQUATIC TOXICOLOGY
·
2016
·
IF 3.557
Jieyi Li,Jinfeng Liu,Yuhuan Zhang,Xuedong Wang,Weijun Li,Hongqin Zhang,Huili Wang
The toxicity of β-diketone antibiotics (DKAs) to larval and adult zebrafish ( Danio rerio ) was investigated by miRNA sequencing and bioinformatics analyses. In control and DKA-exposed groups, 215 differentially expressed miRNAs were screened, and 4076 differential target genes were predicted. Among 51 co-differentially expressed genes, 45 were annotated in KOG functional classification, and 34 in KEGG pathway analysis. The homology analysis of 20 miRNAs with human hsa-miRNAs demonstrated 17 high homologous sequences. The expression levels of 12 miRNAs by qRT-PCR were consistent with those by sRNA-seq. A regulatory network for 4 positive miRNA genes (dre-miR-10, ⿿96, ⿿92 and ⿿184) was plotted, and the high-degree of connectivity between miRNA-gene pairs suggests that these miRNAs play critical roles during zebrafish development. The consistent expression of dre-miR-184 and dre-miR-96 was proved in 120-hpf zebrafish brain, gill, otoliths and lateral line neuromast by qRT-PCR, miRNA-seq, W-ISH and ISH. DKA-exposure led to vacuolation of interstitial cells, reduced number of neurons, glial cell proliferation and formation of glial scar, and the obvious abnormality of cell structure might result from abnormal expression of differentially expressed miRNAs. In general, chronic DKA-exposure resulted in comprehensively toxic effects on larval and adult zebrafish tissues, especially for nervous system.
研究领域:
水生毒理学
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分子生物学
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生物信息学
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产品关联
DOI
PubMed
高影响因子
American Journal of Translational Research
·
2021
·Vol. 13
·期 4
·2111-2126
·
IF 4.06
Chao Shi,Chongwen Shen,Ge Liu,Shaofeng Yang,Fenglin Ye,Jinjin Meng,Youmin Pan
Purpose To clarify the regulatory effect of Nuclear-enriched abundant transcript 1 (NEAT1) on abdominal aortic aneurysm (AAA) model rats and isolated endothelial progenitor cells (EPCs). Methods The AAA rat model was established by CaCl 2 stimulation, and overexpressed NEAT1 was injected into rats through tail vein. Abdominal aorta lesions and numbers of EPCs in tissues and peripheral blood were examined by hematoxylin-eosin, immunofluorescence and flow cytometry. The extracted EPCs were identified by microscopy, DiI-ac-LDL staining and flow cytometry. Effect of overexpressed/silencing NEAT1 on the viability, migration, tube formation and VEGF content of EPCs was investigated by MTT-, wound-healing, tube formation assays and ELISA, respectively. The expressions of NEAT1, miR-204-5p, Angiopoietin-1 (Ang-1)/ERK pathway were determined by qRT-PCR and Western blot as needed. The targeting relationships between NEAT1 and miR-204-5p, and miR-204-5p and Ang-1 were predicted on starBase, TargetScan and confirmed by dual-luciferase experiments. The mutual regulation effect was studied through rescue experiments. Results Overexpressed NEAT1 not only reduced inflammatory infiltration and increased the number of EPCs in abdominal aorta and peripheral blood, but also promoted the viability, migration, tube formation of EPCs, increased VEGF content and upregulated the expression of the Ang-1/ERK pathway in EPCs. However, silencing NEAT1 produced opposite results. NEAT1 targeting miR-204-5p inhibited the functional effects of miR-204-5p on of EPCs. Overexpressed/silencing Ang-1 partially reversed the effects of NEAT1 or miR-204-5p on the characteristics of EPCs. Conclusion NEAT1 competitively binds with miR-204-5p and up-regulates Ang-1 expression in EPCs to effectively improve the proliferation, migration and angiogenesis of EPCs.
研究领域:
分子生物学
·
心血管疾病
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细胞生物学
·
基因调控
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产品关联
DOI
PubMed
高影响因子
RUSSIAN JOURNAL OF PLANT PHYSIOLOGY
·
2018
·Vol. 65
·期 3
·372-383
·
IF 0.816
Zhang Y.,Yong Y. B.,Wang Q.,Lu Y. M.
Lily is regarded as one kind of the most economically important cut flowers in China. In this paper, lily cultivar ‘Aladdin’ ( Lilium longiflorum × Lilium asiatic ) was chosen as experimental material for its distinct high propagation coefficient in nature and three main conclusions were achieved. Firstly, our observation results revealed that the noticeable propagation coefficient of ‘Aladdin’ depended on strong capacity to form many underground stem axillary bulbils positioned on underground node of plant after 70 days of cultivation and having 1 cm flower buds. Secondly, in terms of physiological changes, zeatin riboside (ZR) relative concentration, soluble sugar and starch content peak when and where such bulbils are forming. Finally, two KNOX family Lilium genes were sequenced completely and one BELL family gene was sequenced partly; besides, there might be an interaction between KNOX-2 and BELL protein to accomplish their functions in bulbils organogenesis according to qRT-PCR results. As far as we know, it is the first time to study the formation mechanism of lily underground stem axillary bulbils. We illustrated physiological and molecular profiles, as well as offered a new prospect to lily molecular breeding in the future.
研究领域:
植物学
·
分子生物学
·
植物生理学
·
遗传学
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DOI
高影响因子
ANALYTICAL BIOCHEMISTRY
·
2023
·
IF 2.9
Jianhua Yan,Jiayu Wang,Hongjie Liu,Liwei Wang,Kefu Yu,Li Deng,Junyou Su,Hongfei Chen
Gestational diabetes mellitus (GDM) is a severe perinatal condition with serious consequences for the growth and development of the mother and baby. MicroRNA-29b (miR-29b) is essential to the pathogenesis of GDM and can be used as a molecular biomarker for diagnosis. Given the limitations of current GDM screening technologies, there is a pressing need for a sensitive detection approach to evaluate serum miR-29b in GDM patients, thus aiding in disease treatment. In this study, an electrochemical biosensor Co 7 Fe 3 –CN nanoparticles (NPs) was developed. Using a duplex-specific nuclease (DSN) signal amplification strategy with a linear range of 1–10 4 pM and a low detection limit of 0.79 pM, the ultra-sensitive detection and quantification of miR-29b were accomplished. The dependability and applicability of the developed biosensor were validated by the standard method of qRT-PCR, and the content of serum miR-29b in GDM patients was shown to be significantly lower than that in the control group ( P = 0.03). Specifically, miR-29b concentrations could be detected from 2.0 to 7.5 and 2.4 to 7.3 pM using qRT-PCR and the biosensor, respectively. These similar results indicated that a biosensor based on miR-29b detection has the potential to be used in the point-of-care testing of GDM patients in clinical practice.
研究领域:
生物传感器
·
糖尿病
·
分子生物学
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纳米技术
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临床诊断
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Food Bioscience
·
2025
·
IF 5.9
Yujia Zhou,Zhiming Li,Xinghui Xie,Xingtao Zhou,Jielun Hu,Shaoping Nie
Gellan is a microbial polysaccharide synthesized by Sphingomonas paucimobilis , and is widely utilized in the food and pharmaceutical industries, as well as in other fields. However, much remains unknown about the key genes involved in the gellan synthesis pathway. In this study, ultraviolet (UV) and atmospheric room temperature plasma (ARTP) were employed to induce mutations in S . paucimobilis ATCC31461 (Sp31461 NC), and ultimately resulting a high-yield mutant strain (Sp31461 UA) with a yield increase of 120 % from 3.6 g/L to 8.3 g/L. The growth curve, monosaccharide composition and molecular weight indicated that mutagenesis accelerated the growth of the strain, but did not affect gellan's basic physicochemical property. A combined analysis of transcriptomic and metabolomic demonstrated that the differentially expressed genes and metabolites resulting from mutations were primarily enriched in pathways related to energy metabolism, among which potential key genes such as pgi , clpB , ycaJ associated with gellan yield, were further verified by quantitative reverse transcription polymerase chain reaction (qRT-PCR) and gene overexpression. Notably, the strain overexpression the pgi gene resulted in a 20 % increase in gellan yield compared to the Sp31461 UA strain, reaching 10.0 g/L. In summary, the energy metabolism pathway in S . paucimobilis is closely linked to gellan biosynthesis, and high-yield strains of gellan can be developed through mutagenesis breeding and the overexpression of key genes identified by transcriptomics and metabolomics. This research offers valuable scientific insights for the industrial production and application of gellan.
研究领域:
微生物学
·
工业生物技术
·
分子遗传学
·
代谢工程
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生物化学
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产品关联
DOI
高影响因子
International Journal of Genomics
·
2023
·
IF 2.9
Yi Zhu,Xiaofeng Tian,Yan Wang,Chengxiang Wang,Naiquan Yang,Lianghong Ying,Hongyan Niu
Vascular smooth muscle cells (VSMCs) are critical elements of the vascular wall and play a crucial role in the genesis and development of atherosclerosis (AS). Increasingly, studies have indicated that long noncoding RNAs (lncRNAs) regulate VSMC proliferation, apoptosis, and other biological processes. Nevertheless, the role of lncRNA NFIA-AS1 (hereinafter referred to as NFIA-AS1) in VSMCs and AS remains unclear. Quantitative real-time PCR (qRT-PCR) was performed to analyze the messenger RNA (mRNA) levels of NFIA-AS1 and miR-125a-3p. CCK-8 and EdU staining were performed to detect VSMC proliferation. VSMC apoptosis was evaluated by flow cytometry. The expression of various proteins was detected using western blotting. The levels of inflammatory cytokines secreted by VSMCs were measured by enzyme linked immunosorbent assay (ELISA). The binding sites of NFIA-AS1 and miR-125a-3p, as well as miR-125a-3p and AKT1, were analyzed using bioinformatics methods and validated using a luciferase reporter assay. The function of NFIA-AS1/miR-125a-3p/AKT1 in VSMCs was clarified through loss- and gain-of-functional experiments. We confirmed that NFIA-AS1 was highly expressed in AS tissues and VSMCs induced by oxidized low-density lipoprotein (Ox-LDL). Knockdown of NFIA-AS1 restrained the exceptional growth of Ox-LDL-induced VSMCs, promoted their apoptosis, and decreased the secretion of inflammatory factors and expression of adhesion factors. In addition, NFIA-AS1 regulated the proliferation, apoptosis, and inflammatory response of VSMCs through the miR-125a-3p/AKT1 axis, suggesting that NFIA-AS1 may be a potential therapeutic target for AS.
研究领域:
心血管疾病
·
分子生物学
·
细胞生物学
·
病理学
✓ 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）