ChemistryOpen
·
2019
·Vol. 8
·期 12
·1437-1440
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IF 2.205
Yu-Xin Yue,Yong Kong,Fan Yang,Zhe Zheng,Xin-Yue Hu,Dong-Sheng Guo
Graphical Keeping an eye on enzymes : We designed a supramolecular tandem assay based on the guanidinocalix[5]arene and fluorescein reporter pair, which realized quantitative detection of pyridoxal-5′-phosphate. Guanidinocalix[5]arene and fluorescein reporter pair has been chosen to set up a supramolecular tandem assay principle based on the differential recognition of pyridoxal-5′-phosphate (the substrate of alkaline phosphatase, ALP), pyridoxal (the product of ALP) and phosphate (the product of ALP). This supramolecular tandem assay system offers an opportunity to monitor the activity of ALP in a label-free, continuous, and real-time manner. More importantly, a calibration curve can be given for selective and quantitative detection of pyridoxal-5′-phosphate (biomarker for several diseases).
研究领域:
分析化学
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生物化学
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分子生物学
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医学
✓ 已验证
DOI
PubMed
高影响因子
TALANTA
·
2017
·
IF 4.162
Yaqi Mei,Qiong Hu,Baojing Zhou,Yonghui Zhang,Minhui He,Ting Xu,Feng Li,Jinming Kong
Simple and fast detection of alkaline phosphatase (ALP) activity is of great importance for diagnostic and analytical applications. In this work, we report a turn-off approach for the real-time detection of ALP activity on the basis of the charge transfer induced fluorescence quenching of the Cu(BCDS) 2 2- (BCDS = bathocuproine disulfonate) probe. Initially, ALP can enzymatically hydrolyze the substrate ascorbic acid 2-phosphate to release ascorbic acid (AA). Subsequently, the AA-mediated reduction of the Cu(BCDS) 2 2- probe, which displays an intense photoluminescence band at the wavelength of 402 nm, leads to the static quenching of fluorescence of the probe as a result of charge transfer. The underlying mechanism of the fluorescence quenching was demonstrated by quantum mechanical calculations. The Cu(BCDS) 2 2- probe features a large Stokes shift (86 nm) and is highly immune to photo bleaching. In addition, this approach is free of elaborately designed fluorescent probes and allows the detection of ALP activity in a real-time manner. Under optimal conditions, it provides a fast and sensitive detection of ALP activity within the dynamic range of 0–220 mU mL −1 , with a detection limit down to 0.27 mU mL −1 . Results demonstrate that it is highly selective, and applicable to the screening of ALP inhibitors in drug discovery. More importantly, it shows a good analytical performance for the direct detection of the endogenous ALP levels of undiluted human serum and even whole blood samples. Therefore, the proposed charge transfer based approach has great potential in diagnostic and analytical applications.
研究领域:
分析化学
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生物化学
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分子诊断
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药物发现
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Materials Science & Engineering C-Materials for Biological Applications
·
2021
·
IF 7.328
Qifan Wang,Wenjie Ye,Zhiyong Ma,Wenjia Xie,Linna Zhong,Ying Wang,Qiong Rong
Scaffolds prepared by 3D printing are increasingly used in the field of bone tissue repair. However, on traditional 3D printed bone tissue engineering scaffolds, cells can only grow on the fiber surface and form bone. We designed a scaffold with a cross-scale structure of PCL/β-TCP, which contains thick fibers with a diameter of 500 μm printed by FDM. And in the pores of the coarse fiber, the ultra-high precision fine fiber grid with a diameter of about 10 μm is filled by MEW mode. In cell experiments, cells can not only grow on the thick fiber surface of the cross-scale scaffold. At the same time, the mesh structure of fine fibers provides a bridge for cell growth, allowing cells to pass through the pores of thick fibers and grow in the pores and gradually cover the pores of the scaffold. In the osteoinduction experiment, β-TCP in the PCL/β-TCP composite provides Ca 2+ and PO 4 3− to the scaffold, which effectively promotes the osteogenic differentiation of cells on the scaffold. Compared with traditional scaffolds, the osteogenic performance of cross-scale scaffolds is greatly improved. Not only did bone form on the surface of the scaffold, but also obvious ALP expression and effective calcium precipitation appeared in the pores of the scaffold. This can effectively speed up the repair of bone defects. We believe that the 3D printed PCL/β-TCP cross-scale scaffold with high-precision fibers has great application prospects in the field of bone tissue engineering.
研究领域:
生物医学工程
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组织工程
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材料科学
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骨科
✓ 已验证
产品关联
DOI
PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2016
·
IF 7.476
Huifang Zhang,Xiaoming Ma,Shuisheng Hu,Yue Lin,Longhua Guo,Bin Qiu,Zhenyu Lin,Guonan Chen
Development of convenient but sensitive method for influenza detection is highly important in immediate and effective clinical treatment. In this study, an ultrasensitive colorimetric approach combining the advantages of the convenience of the enzyme-induced metallization and the high specificity of enzyme-linked immunosorbent assay for the detection of influenza virus A (H7N9 as model) has been developed. Two rounds of amplification are utilized to enhance the detection sensitivity. The amplification of enzymatic reaction combines with the specific optical properties of gold nanoparticles causing the enhancing of the optical signal immensely. In addition, the increased surface area and the magnetic enrichment effect also enable the magnetic bead (MB) to catch a large number of alkaline phosphatase (ALP) and detection antibody (Ab 2 ), thus very small amounts of the virus can be easily detected. Compared with conventional method, this approach exhibits outstanding sensitivity for ALP detection, 0.2 U/L of ALP can be distinguished with a spectrometer and 2 U/L with the naked eye. And as low as 25 pg/mL of H7N9 can be detected by the naked eye. This approach shows an extensive horizon for bioassays and is available in clinical diagnosis with the advances of simplification, effectiveness, low cost and sensitive readout.
研究领域:
病毒学
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诊断医学
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纳米技术
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分析化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Nutrients
·
2022
·Vol. 14
·期 9
·1940
·
IF 6.706
Kuaitian Wang,Xiao Kong,Mengdi Du,Wei Yu,Zhenhua Wang,Bo Xu,Jianrong Yang,Jingru Xu,Zhili Liu,Yongqiang Cheng,Jing Gan
DEDEQIPSHPPR, the calcium-binding peptide (CBP) identified in soy yogurt, was proven to be a potential cofactor in osteoporosis prevention in our previous study, but the mechanism was unknown. In this study, the activity of alkaline phosphatase (ALP) and osteocalcin (OCN), the regulation of RUNX2, and the expression of TβRI were investigated to elucidate the underlying mechanism. The results show that CBP upregulated ALP activity and OCN concentration and increased the expression of RUNX2 and the activation of the MAPK signaling pathway. Similarly, the expression of osteogenesis-related genes in osteoblasts also increased upon CBP treatment. Moreover, the CBP-induced enhancement of ALP activity and phosphorylation levels in the p38 pathway was inhibited by treatment with a p38 inhibitor (SB203538) and TβRI inhibitor (SB431542), respectively, suggesting that p38 and TβRI were involved in the osteogenic action. Based on the signaling pathways, the intracellular calcium concentration was significantly elevated by CBP, which was correlated with the increased behavioral functions and the relative fluorescence intensity of the bone mass. These findings suggest that CBP stimulates osteoblast differentiation and bone mineralization through the activation of RUNX2 via mechanisms related to the TβRI-p38-MAPK signaling pathways, further highlighting CBP’s important potential for treating osteoporosis.Keywords:bone remodeling;soy peptide CBP;osteoblast;differentiation;p38-MAPK;RUNX2
研究领域:
生物学
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生物化学
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药理学
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骨骼生物学
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产品关联
DOI
PubMed
高影响因子
RSC Advances
·
2019
·Vol. 9
·期 66
·38590-38596
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IF 3.049
Rong Huang,Lu Lu Xiong,Hui Hui Chai,Jing Jing Fu,Zhisong Lu,Ling Yu
A novel colorimetric aptasensor based on a Au/Fe3O4 nanohybrid was developed to detect ochratoxin A (OTA). The aptasensor is composed of a free OTA aptamer, a Au/Fe3O4 nanohybrid coated with biotinylated complementary DNA of the OTA aptamer (biotin-cDNA-Au/Fe3O4), and free alkaline-phosphatase-labeled streptavidin (SA-ALP). The Au/Fe3O4 nanohybrid not only immobilizes biotin-cDNA but also magnetically separates SA-ALP from the sample solution. One part of the OTA aptamer sequence hybridizes with biotin-cDNA immobilized on Au/Fe3O4, and the left part of the OTA aptamer sequence covers the biotin and blocks the specific interaction between biotin and SA-ALP. OTA can interrupt the interaction of OTA aptamer binding to biotin-cDNA-Au/Fe3O4 and can inhibit the shielding effect of the OTA aptamer on biotin. The amount of SA-ALP that can be captured by biotin-cDNA-Au/Fe3O4 thus increases with increasing OTA concentration. Through a simple magnetic separation, the collected SA-ALP-linked Au/Fe3O4 can produce a yellow-colored solution in the presence of p-nitrophenyl phosphate (p-NPP). This colorimetric aptasensor can detect OTA as low as 1.15 ng mL−1 with high specificity.
研究领域:
分析化学
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纳米技术
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毒理学
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食品安全
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Nanoscale
·
2018
·Vol. 10
·期 15
·7163-7170
·
IF 7.233
Jiahui Zhao,Shuang Wang,Shasha Lu,Jian Sun,Xiurong Yang
As a ubiquitous hydrolysis enzyme in phosphate metabolism, alkaline phosphatase (ALP) is a significant biomarker in laboratory research and clinic diagnosis. Herein, we report a highly water-soluble Eu(DPA)3@Lap nanohybrid material for the rapid and selective assay of PPi and ALP through a luminescence off–on recognition process. Eu(DPA)3@Lap was successfully prepared in an aqueous solution, and it exhibited strong luminescence emission, high photostability, and long lifetime. More interestingly, the strong luminescence of Eu(DPA)3@Lap can be remarkably quenched by Cu2+ due to the high impetus of coordination between the DPA ligand and Cu2+ ion. Using Cu2+ as a signal transducer, the luminescence could be recovered upon the addition of PPi ion owing to the formation of a Cu2+–PPi complex; thus, a luminescence turn-on assay for PPi ions was realized. Utilizing the ability of Cu2+ to differentiate between PPi and Pi, a convenient and straightforward luminescence assay for ALP activity was accomplished based on the specific dephosphorylation of PPi to Pi. To the best of our knowledge, this elaborate luminescence sensing system constitutes the first luminescent nanohybrid material based on a europium organic complex for ALP activity assay. Furthermore, the recognition process of PPi and ALP was completed in a convenient and facile mix-and-readout manner, and it revealed significant potential in point of care testing.
研究领域:
生物化学
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分析化学
·
材料科学
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医学诊断
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Science China-Chemistry
·
2023
·
IF 9.6
Tang Xianqing,Chen Jinghuang,Zhang Mengqian,Sun Jian,Yang Xiurong
Alkaline phosphatase (ALP) activity and pyrophosphate ion (PPi) levels are remarkable for the human body functions such as signal transduction pathways and metabolism. Current quantitative methods mainly focus on developing complicated organic substrates or employing unstable metal ions as signal-regulated medium. Herein, we have developed a facile hydrothermal method for preparing FeWO 4 nanomaterials with intrinsic peroxidase-like activity and further confirmed that such a catalytic activity could be significantly enhanced by adjusting the size and oxygen vacancy content. More encouragingly, PPi can easily inhibit the catalytic activity of FeWO 4 , whereas orthophosphate ions (Pi) cannot. Therefore, we constructed an FeWO 4 -based colorimetric assay for sensing PPi by means of the classical 3,3′,5,5′-tetramethylbenzidine-peroxidase chromogenic reaction. A facile and reliable ALP activity assay was also designed and developed because of the logical regulation of the peroxidase-like activity of FeWO 4 through the ALP-catalyzed hydrolysis of PPi into Pi. Based on the clear mechanism and mimetic-enzyme FeWO 4 -catalyzed amplification, the sensing system exhibited excellent performance and was able to evaluate ALP activity in real serum samples and screen for potential ALP inhibitors. The proposed mimetic enzyme-involved colorimetric assay provides an alternative pathway, and FeWO 4 nanomaterials with excellent performance have great potential for further biosensing and biomedical applications.
研究领域:
纳米技术
·
生物化学
·
催化
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分析化学
✓ 已验证
产品关联
DOI
高影响因子
Frontiers in Pharmacology
·
2024
·
IF 4.4
Xiaoqin Zhang,Jiale Mao,Lu Shao,Shuang Liu,Jiwang Zhou,Mingrong Mei,Zunjing Zhang
Background: Melastoma dodecandrum Lour. (MD), a traditional botanical drug known for its hypoglycemic, antioxidant, and anti-inflammatory properties, is commonly used to treat diabetes, osteoarthritis, and osteoporosis. However, its specific active components against diabetic osteoporosis remain unclear.Purpose: This study aimed to identify the key active components in MD using cell membrane chromatography coupled with mass spectrometry and validate their effects in vitro.Methods: An AGEs-induced osteoblast injury model was established. MTT assays measured cell viability, and ALP activity was assessed using a biochemical kit. Western blotting was employed to detect the expression levels of osteoblast-related proteins OCN and RUNX2 and the AGE receptor protein RAGE. ELISA was used to determine the levels of SOD, MDA, CAT, and GPx. PCR quantified TNF-α expression to evaluate the protective effects and potential mechanisms of MD. The AGEs-induced osteoblast cell membrane chromatography-mass spectrometry method facilitated the rapid identification of potentially active compounds based on their affinity for the osteoblast cell membrane. Cell experiments further validated the activity of the characteristic component isovitexin.Results: MD significantly improved cell viability in AGEs-damaged osteoblasts, enhanced ALP, SOD, CAT, and GPx activities, reduced MDA levels, increased OCN and RUNX2 protein expression, and decreased TNF-α mRNA and RAGE protein expression. Cell membrane chromatography identified 20 chemical constituents, including 13 flavonoids, 4 organic acids, 1 phenylpropanoids, 1 terpenoids, and 1 alkaloid. Cell experiments have confirmed that isovitexin has significant protective activity against osteoblasts and can inhibit the expression of specific receptor RAGE on the osteoblast membrane, consistent with the effect of MD.Conclusion: MD and its active component, isovitexin, provide protective effects against AGEs-induced osteoblast injury, offering a basis for subsequent drug development.
研究领域:
药理学
·
生物化学
·
细胞生物学
·
分析化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2017
·
IF 5.401
Juan Ou-Yang,Chun-Yan Li,Yong-Fei Li,Bin Yang,Song-Jiao Li
Alkaline phosphatase (ALP) is an essential enzyme in phosphate metabolism and widely distributes in mammalian body fluids and tissues. So far, various fluorescent probes for ALP have been well developed, but some disadvantages such as short emission wavelength and low photostability restrict their biological application. Herein, a near-infrared (NIR) fluorescent nanoprobe based on infinite coordination polymer nanoparticles (CyOH@Tb-GMP) is constructed for endogenous ALP activity assay. The characteristics of the nanoprobe are as following: (1) The fluorescence emission of the nanoprobe is at 718 nm belonging to NIR region, which is suitable for bioimaging. (2) The nanoprobe displays better photostability than common NIR dyes such as ICG as well as CyOH. (3) The nanoprobe exhibits high sensitivity for ALP activity with a 7.5-fold fluorescence enhancement when 2.5 U/mL ALP is added. (4) The NIR nanoprobe is employed for monitoring endogenous ALP activity in various biological samples such as cell, tissue and mice with satisfactory results.
研究领域:
生物化学
·
分子生物学
·
纳米技术
·
医学影像
✓ 已验证
产品关联
DOI
高影响因子
Tissue Engineering and Regenerative Medicine
·
2023
·
IF 3.6
Zhang Ying,Li Chuan,Wei Qiushi,Yuan Qiang,He Wei,Zhang Ning,Dong Yiping,Jing Zhenhao,Zhang Leilei,Wang Haibin,Cao Xiangyang
BACKGROUND: This study aimed to explore the effect of a nanomaterial-based miR-320a inhibitor sustained release system in trauma-induced osteonecrosis of the femoral head (TIONFH). METHODS: The miR-320a inhibitor-loaded polyethylene glycol (PEG)- Poly(lactic-co-glycolic acid) (PLGA)- Poly-L-lysine (PLL) nanoparticles were constructed using the double emulsion method. The TIONFH rabbit model was established to observe the effects of miR-320a inhibitor nanoparticles in vivo . Hematoxylin–eosin staining and microcomputed tomography scanning were used for bone morphology analysis. Bone marrow mesenchymal stem cells (BMSCs), derived from TIONFH rabbits, were used for in vitro experiments. Cell viability was determined using the MTT assay. RESULTS: High expression of miR-320a inhibited the osteogenic differentiation capacity of BMSCs in vitro by inhibiting the expression of the osteoblastic differentiation markers ALP and RUNX2. MiR-320a inhibitor-loaded PEG-PLGA-PLL nanoparticles were constructed with a mean loading efficiency of 1.414 ± 0.160%, and a mean encapsulation efficiency of 93.45 ± 1.24%, which released 50% of the loaded miR-320a inhibitor at day 12 and 80% on day 18. Then, inhibitor release entered the plateau. After treatment with the miR-320a inhibitor nanoparticle, the empty lacunae were decreased in the femoral head tissue of TIONFH rabbits, and the osteoblast surface/bone surface (Ob.S/BS), osteoblast number/bone perimeter (Ob.N/B.Pm), bone volume fraction, and bone mineral density increased. Additionally, the expression of osteogenic markers RUNX2 and ALP was significantly elevated in the TIONFH rabbit model. CONCLUSION: The miR-320a inhibitor-loaded PEG-PLGA-PLL nanoparticle sustained drug release system significantly contributed to bone regeneration in the TIONFH rabbit model, which might be a promising strategy for the treatment of TIONFH.
研究领域:
纳米技术
·
骨科
·
再生医学
·
分子生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
JOURNAL OF RARE EARTHS
·
2025
·
IF 7.2
Yang Lu,Xinhua Dai,Yidi Wang,Ting Liu,Chao Liu,Xu Yan,Xiaomin Liu,Bai Li
Alkaline phosphatase (ALP), a hydrolase commonly found in living organisms, shows significantly elevated levels in serum of patients suffering from liver metastasis. Rapid and efficient detection of ALP activity in serum is thus critical for assessing cancer prognosis. In this study, we developed a composite nanoprobe (UCNPs@ZIF@PDA) based on upconversion nanoparticles (UCNPs), specifically NaErF 4 :Tm (0.5 mol%)@NaLuF 4 , characterized by strongly correlated energy levels, for the sensitive and stable detection of ALP in complex environments. To improve stability, UCNPs were encapsulated with zeolitic imidazolate framework-8 (ZIF-8). The upconversion luminescence (UCL) of the UCNPs was subsequently quenched by polydopamine (PDA), which was formed via in-situ polymerization of dopamine on the ZIF-8 surface. ALP catalyzed the conversion of L-ascorbic acid trisodium 2-phosphate (AAP) to ascorbic acid, which in turn triggered the degradation of PDA and restored the UCL. Through this mechanism, the limits of detection for ALP in aqueous solution and serum are 0.19 and 0.42 U/L, respectively. Moreover, the probe demonstrates precise imaging of ALP in cellular environments and successfully distinguishes ALP levels in samples with and without liver metastasis. These findings highlight the strong potential of UCNP-based composite fluorescent probes as a novel and efficient tool for ALP detection in clinical applications.
研究领域:
生物医学工程
·
纳米技术
·
生物化学
·
临床诊断
·
分子成像
✓ 已验证
产品关联
DOI
高影响因子
危险品化学品经营许可证（带存储）