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ANALYTICA CHIMICA ACTA · 2024 · IF 5.7

Pyrroloquinoline quinone exhibiting peroxidase-mimicking property for colorimetric assays

Ning Xia,Fengli Gao,Gang Liu,Yong Chang,Lin Liu

Background Small molecule mimics offer the advantages of easy preparation, good thermodynamic stability, and reproducible catalytic activity. However, most of the reported organic artificial mimics face challenges including low catalytic activity, oxidative self-destruction, and auto-aggregation into inactive dimers. Therefore, novel organic mimics with high catalytic activity as well as good thermal and environmental stability are highly desirable. Results We found that pyrroloquinoline quinone (PQQ), a nutritionally important growth factor, displayed significant peroxidase-like activity to catalyze the oxidation of colorless 3,3′,5,5′-tetramethylbenzidine (TMB) into blue oxidized TMB (oxTMB) by H 2 O 2 . Its catalytic activity surpassed that of other organic small molecules reported previously, including fluorescein derivatives and adenosine/guanosine triphosphate. Based on the reduction of oxTMB by the enzymatically generated ascorbic acid (AA), the PQQ/TMB-based system was used to monitor alkaline phosphatase (ALP) activity with ascorbic acid 2−phosphate (AAP) as the substrate. This sensing system allowed for the determination of ALP with a detection limit of 1 mU/mL. Furthermore, we discovered that PQQ can coordinate with Cu 2+ to form metal-organic nanoparticles. The building blocks of two active cofactors endowed the nanoparticles with impressive functionalities for biocatalytic applications. With Cu-PQQ nanoparticles as signal labels, colorimetric immunoassays of prostate specific antigen (PSA) were achieved with a lowest detectable concentration down to 0.1 pg/mL. Significance This work is valuable for the design of novel biosensors by utilizing PQQ as an artificial enzyme because of its high catalytic activity, good stability, ease of storage, and simple chemical structure.

研究领域: 生物化学 · 分析化学 · 纳米技术 · 生物传感器
✓ 已验证 产品关联 DOI PubMed 高影响因子
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LAB ON A CHIP · 2016 ·Vol. 17 ·期 1 ·120-127 · IF 5.586

Microfluidic PDMS on paper (POP) devices

Jin-Wen Shangguan,Yu Liu,Jian-Bin Pan,Bi-Yi Xu,Jing-Juan Xu,Hong-Yuan Chen

In this paper, we propose a generalized concept of microfluidic polydimethylsiloxane (PDMS) on paper (POP) devices, which combines well the merits of paper chips and PDMS chips. First, we optimized the conditions for accurate PDMS spatial patterning on paper, based on screen printing and a high temperature enabled superfast curing technique, which enables PDMS patterning to an accuracy of tens of microns in less than ten seconds. This, in turn, makes it available for seamless, reversible and reliable integration of the resulting paper layer with other PDMS channel structures. The integrated POP devices allow for both porous paper and smooth channels to be spatially defined on the devices, greatly extending the flexibility for designers to be able to construct powerful functional structures. To demonstrate the versatility of this design, a prototype POP device for the colorimetric analysis of liver function markers, serum protein, alkaline phosphatase (ALP) and aspartate aminotransferase (AST), was constructed. On this POP device, quantitative sample loading, mixing and multiplex analysis have all been realized.

CAS / MDL: 328-50-7
研究领域: 微流控技术 · 生物医学工程 · 分析化学
✓ 已验证 产品关联 DOI PubMed 高影响因子
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ELECTROCHIMICA ACTA · 2019 · IF 5.383

Full-spectrum responsive photoelectrochemical immunoassay based on β-In2S3@carbon dot nanoflowers

Zhonghua Yu,Lingting Huang,Jialun Chen,Ying Tang,Biyun Xia,Dianping Tang

This work designed a new photoelectrochemical (PEC) immunoassay for the ultrasensitive detection of alpha-fetoprotein (AFP) based on the flower-like β-In 2 S 3 @carbon dots (CDs). The photocurrent of the as-prepared heterogeneous nanostructure was enhanced significantly under ultraviolet (UV), visible (Vis) and near-infrared (NIR) irradiation thanks to the more generation of carriers and less electron-hole pairs recombination with CDs. The three-layer sandwich-like immunocomplex would be formed upon the target AFP existed, the ascorbic acid 2-phosphate (AA2P) was subsequently transformed into ascorbic acid (AA) owing to the catalysis of alkaline phosphatase-conjugated rabbit polyclonal anti-human AFP (ALP-Ab2). Without additional applied voltage, the photocurrent of the β-In 2 S 3 @CDs-modified electrode was positively correlated with the increasing AA level in the detection cell. Under optimal conditions, the developed PEC immunosensing platform exhibited good reproducibility and high sensitivity for the determination of target AFP within a dynamic working range from 0.5 to 50 ng mL −1  at a detection limit of 37.9 pg mL −1 . In addition, the method accuracy relative to human AFP ELISA kit was evaluated by analyzing human serum specimens, and a good match between two methods was obtained. Such the increasing photocurrent through full-spectrum responsive mode gives a new horizon to develop highly sensitive PEC immunoassay in early clinical diagnosis by monitoring disease-related biomarkers.

CAS / MDL: 107-15-3 22519-64-8
研究领域: 分析化学 · 生物医学工程 · 临床诊断
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ANALYTICA CHIMICA ACTA · 2022 · IF 6.911

Integrated enzyme with stimuli-responsive coordination polymer for personal glucose meter-based portable immunoassay

Liping Gao,Yong Li,Zhen-Zhong Huang,Hongliang Tan

Coordination polymers (CPs) with tunable structures and properties have been extensively explored in a variety of fields. In this work, we demonstrated the potential of stimuli-responsive CPs as a host of integrating enzymes to construct a portable immunoassay . By employing terbium ion (Tb 3+ ) as a metal node and guanine monophosphate (GMP) as a bridge ligand, an alkaline phosphatase (ALP)-responsive Tb/GMP CPs was fabricated, which allows amyloglucosidase (GA) to be integrated to form [email protected] /GMP composite. Owing to the size-selectivity of Tb/GMP CPs as a host, the loaded GA was physically isolated from its substrate starch. However, Tb/GMP CPs is highly sensitive to ALP, which can hydrolyze the phosphate group of GMP to destroy the structure of Tb/GMP CPs, leading to the release of GA from [email protected] /GMP composite. The released GA can digest starch to produce glucoses and give a measured signal by personal glucose meter (PGM). This finding leads to a PGM-based portable immunoassay for the quantitative analysis of carcinoembryonic antigen (CEA), and satisfactory results with a detection limit of 0.28 ng/mL have been achieved. The successful determination of CEA in serum samples demonstrates its potential in practical application. We believe that this work can provide a remarkable insight for the rational design of stimuli-responsive CPs for a wide of applications.

研究领域: 材料科学 · 分析化学 · 生物化学 · 医学诊断
✓ 已验证 产品关联 DOI PubMed 高影响因子
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JOURNAL OF APPLIED ELECTROCHEMISTRY · 2022 ·Vol. 53 ·期 3 ·547-557 · IF 2.925

Specific unlocking of the butterfly effect: nanointerface-based electrochemical biosensing of adenosine triphosphate and alkaline phosphatase

Hu Kaiyue,Ren Xinxin,Qin Lingxia,Guo Zhiyong,Wu Di,Wang Sui,Hu Yufang

The purpose of this study was to achieve a specific unlocking of the butterfly effect: nanointerface-based electrochemical biosensing of adenosine triphosphate (ATP) and alkaline phosphatase (ALP). Based on the Faraday-cage concept reported first by our group, we built a new outer Helmholtz plane (OHP)-based electrochemical biosensor by using an unique nanocomposite involving three-dimensional graphene-Au nanoparticles (3D-GO-AuNPs), tetrahedral DNA nanostructures (TDNs), and separated ATP aptamers, in which methylene blue (MB) was employed as the electrochemical signal output. In this process, the prepared nanocomposites were attached favorably onto the TDN substrate electrode surface due to the interaction of ATP and its aptamer, creating a better OHP of the electrode owing to its large enough specific surface area; then a detection limit of 0.25 pM was calculated by 3 δ /slope. Whereas, the hydrolysis for ATP of ALP can hinder this binding process, therefore, the biosensor could be indirectly applied for ALP analysis with a detection limit of 0.21 mU/L (3 δ /slope). Since some small changes of the two targets will set off a whole series of changes in system, the OHP-extended biosensor provides a superior electrochemical platform for complex biological processes with causal relationships in clinical diagnosis and drug development, similar to the butterfly effect. Graphical abstract

研究领域: 生物化学 · 纳米技术 · 电化学 · 临床诊断
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ANALYTICAL CHEMISTRY · 2023 · IF 7.4

MnO4–-Triggered Immediate-Stable Real-Time Fluorescence Immunosensor with High Response Speed and Low Steady-State Error

Wenlin Ma,Lihua Pang,Jinhua Liu,Lei Wen,Huili Ma,Yinhui Li,Zhihui Xu,Chengwu Zhang,Hai-Dong Yu

Real-time chemical and biological sensing in vitro is important for application in health and environmental monitoring. Thus, a more rapid and stable detection method is urgently needed. Herein, an immediate-stable real-time fluorescent immunosensor with a high response speed (∼100%, <1 s) and approximately zero steady-state error is constructed. The developed sensor is based on the MnO4–-triggered in situ immediate-stable fluorogenic reaction between dopamine and orcinol monohydrate to produce azamonardine (DMTM). The obtained DMTM is identified and characterized by high-resolution mass spectrometry, 1H NMR spectroscopy, 13C NMR spectroscopy, and theoretical calculations. The present sensor achieves a highly sensitive detection of dopamine (DA) with a limit of detection (LOD) of 10 nM as well as alkaline phosphates (ALP) with an LOD of 0.1 mU/mL by using orcinol monohydrate phosphate sodium salt as a substrate. As a proof of concept, ALP-triggered fluorescence ELISA using cardiac troponin I (cTnI) as a model antigen target is further constructed. The developed real-time sensor achieves the detection of cTnI with an LOD of 0.05 ng/mL. Moreover, the sensor proposed by us is successfully applied to assess the cTnI level in clinical serum specimens and yields results consistent with those obtained by the commercial ELISA method. The immediate-stable real-time fluorescence immunosensor provides a promising and powerful platform for the trace detection of biomolecules in clinical diagnosis.

研究领域: 分析化学 · 生物化学 · 医学诊断 · 环境监测 · 临床化学
✓ 已验证 产品关联 DOI PubMed 高影响因子
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Small · 2023 · IF 13.3

Magnetic Field Boosts the Transmembrane Transport Efficiency of Magnesium Ions from PLLA Bone Scaffold

Zuyun Yan,Tianshi Sun,Wei Tan,Zhicheng Wang,Jinpeng Yan,Jinglei Miao,Xin Wu,Pei Feng,Youwen Deng

In the system of magnesium-loaded scaffolds, the effect of magnesium ions (Mg 2+ ) on the osteogenesis induction is restricted due to the low transmembrane transport efficiency of Mg 2+ into the cell, which limits the application for bone defect repair. Inspired by the fact that magnetic field can regulate ion channel proteins on the cell membrane, magnetite nanoparticle is introduced into the poly (l-lactic acid) /magnesium oxide composite in this study, and a magnetic magnesium-loaded bone scaffold is prepared via selective laser sintering . Notably, the activities of the Mg 2+ channel protein (MAGT1) on the membrane of bone marrow mesenchymal stem cells (rBMSCs) are enhanced via magnetic torque effect (via integrin α V β 3/actin), under the action of static magnetic field (SMF), which promoted rBMSCs to capture Mg 2+ in the microenvironment and induced osteogenesis. In vitro experiments showed that the magnetic magnesium-loaded scaffold, under the action of SMF, can accelerate the inflow of Mg 2+ from surrounding microenvironment, which improved cellular activities, osteogenesis-related gene expression (ALP, Runx2, OCN, and OPN), and mineralization. Besides, in vivo skull defect repair experiments showed that the scaffolds possessed good ability to promote bone differentiation and new bone regeneration.

CAS / MDL: 1317-61-9 1309-48-4
研究领域: 生物材料 · 组织工程 · 生物医学工程 · 细胞生物学 · 骨科
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MICROCHIMICA ACTA · 2024 ·Vol. 191 ·期 7 ·1-8 · IF 5.3

Atomically Co-dispersed nitrogen-doped carbon for sensitive electrochemical immunoassay of breast cancer biomarker CA15-3

Han Lei,Cai Shuanglong,Chen Xiaogeng

The development of an ultrasensitive and precise measurement of a breast cancer biomarker (cancer antigen 15-3; CA15-3) in complex human serum is essential for the early diagnosis of cancer in groups of healthy populations and the treatment of patients. However, currently available testing technologies suffer from insufficient sensitivity toward CA15-3, which severely limits early large-scale screening of breast cancer patients. We report a versatile electrochemical immunoassay method based on atomically cobalt-dispersed nitrogen-doped carbon (Co-NC)–modified disposable screen-printed carbon electrode (SPCE) with alkaline phosphatase (ALP) and its metabolite, ascorbic acid 2-phosphate (AAP), as the electrochemical labeling and redox signaling unit for sensitive detection of low-abundance CA15-3. During electrochemical detection by differential pulse voltammetry (DPV), it was found that the Co-NC-SPCE electrode did not have a current signal response to the AAP substrate; however, it had an extremely favorable response current to ascorbic acid (AA). Based on the above principle, the target CA15-3-triggered immunoassay enriched ALP-catalyzed AAP produces a large amount of AA, resulting in a significant change in the system current signal, thereby realizing the highly sensitive detection of CA15-3. Under the optimal AAP substrate concentration and ALP catalysis time, the Co-NC-SPCE-based electrochemical immunoassay demonstrated a good DPV current for CA15-3 in the assay interval of 1.0 mU/mL to 10,000 mU/mL, with a calculated limit of detection of 0.38 mU/mL. Since Co-NC-SPCE has an excellent DPV current response to AA and employs split-type scheme, the constructed electrochemical immunoassay has the merits of high preciseness and anti-interference, and its clinical diagnostic results are comparable to those of commercial kits. Graphical abstract

研究领域: 分析化学 · 生物化学 · 电化学 · 癌症诊断
✓ 已验证 产品关联 DOI PubMed 高影响因子
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BIOMEDICINE & PHARMACOTHERAPY · 2024 · IF 6.9

The bone-protective benefits of kaempferol combined with metformin by regulation of osteogenesis-angiogenesis coupling in OVX rats

Zhongyuan Zhang,Wenshu Xu,Zhenhua Zhang,Xiaoxue Chen,Hui Jin,Ningning Jiang,Hui Xu

This study was to investigate the potential mechanisms of treatment with metformin (Met) combined with kaempferol (Kae) against postmenopausal osteoporosis. Experiments were conducted in both ovariectomy (OVX)-induced osteoporosis rats and in vitro using RAW264.7 cells, MC3T3-E1 cells, and HUVECs. Results demonstrated the therapeutic effect of Met combined with Kae on osteoporosis. In vivo, Kae alone and in combination with Met treatments enhanced tibial trabecular microstructure, bone mineral density (BMD), and mechanical properties in OVX rats without causing hepatotoxicity and nephrotoxicity. It also reduced bone resorption markers (CTX-1 and TRAP) and increased the bone formation marker (PINP) level in the serum of OVX rats. The expression of bone resorption marker TRAP was reduced, while bone formation markers Runx2 and ALP were enhanced in the bone tissue of OVX rats. Furthermore, Met combined with Kae also promoted the expression of angiogenesis-related markers CD31 and VEGF in OVX rats. In vitro, MC3T3-E1s cells treated with Met combined with Kae showed higher expression of ALP, Runx2, and VEGF. Interestingly, the treatment did not directly promote HUVECs migration and angiogenesis, but enhanced osteoblast-mediated angiogenesis by upregulating VEGF levels. Additionally, Met combined with Kae treatment promoted VEGF secretion in MC3T3-E1, and activated the Notch intracelluar pathway by upregulating HES1 and HEY1 in HUVECs. Meantime, their stimulation on CD31 expression were inhibited by DAPT, a Notch signaling inhibitor. Overall, this study demonstrates the positive effects of Met combined with Kae on osteoporotic rats by promoting osteogenesis-angiogenesis coupling, suggesting their potential application in postmenopausal osteoporosis.

研究领域: 药理学 · 骨科 · 分子生物学 · 细胞生物学
✓ 已验证 DOI PubMed 高影响因子
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TALANTA · 2018 · IF 4.244

Construction of liquid crystal droplet-based sensing platform for sensitive detection of organophosphate pesticide

Lele Zhou,Qiongzheng Hu,Qi Kang,Li Yu

Alkaline phosphatase (ALP), as an essential disease biomarker, widely distributes in various tissues, and plays a significant role in diagnosis and treatment of some diseases (e.g. cancers and liver dysfunction). Dichlorvos (DDVP), as one of typical organophosphate pesticide , could be hydrolyzed by ALP. In this work, we constructed a simple but robust 4-cyano-4′-pentylbiphenyl (5CB) droplet sensing platform for sensitive and convenient detection of DDVP based on its hydrolysis by ALP. The optical responses of liquid crystals (LCs) originated from controlling orientations of 5CB by using an ALP cleavable surfactant, sodium monododecyl phosphate (SMP). A dark crossed optical image of 5CB was observed due to formation of the SMP monolayer at the aqueous/LC droplet interface. Subsequently, 5CB turned to bright fan-shaped optical appearance upon addition of ALP and SMP mixture. Interestingly, LCs remained dark crossed optical images in contact with the pre-incubated mixture of DDVP and ALP. The detection limit of DDVP reached ~0.1 ng/mL. Isothermal titration calorimetry (ITC) and humam ALP ELISA kit provided strong evidence for the detection mechanism. This strategy was also used to evaluate whether the maximum pesticide residue limits of tomatoes conform to the relevant national standard requirement or not. Our study holds promise for real-time and label-free detection of pesticides with high sensitivity by employing portable LC-based sensing platform.

CAS / MDL: 62-73-7 9000-90-2
研究领域: 分析化学 · 环境科学 · 生物化学 · 食品安全
✓ 已验证 产品关联 DOI PubMed 高影响因子
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Bioengineered · 2021 · IF 3.269

Down-regulation of miR-340-5p promoted osteogenic differentiation through regulation of runt-related transcription factor-2 (RUNX2) in MC3T3-E1 cells

Xiaochen Wang,Yaochuan Mi,Wei He,Xiaona Hu,Shuo Yang,Lu Zhao,Yanyang Zhang,Binhong Wen

Diabetic osteoporosis (DOP) is a chronic complication of diabetes in the skeletal system. High level of miR-340-5p may be harmful to the bone formation. In this study, the DOP model of rats was successfully established via streptozotocin (STZ) and ovariectomy (OVX) treatment. It was manifested by reduced body weight, insulin level, alkaline phosphatase (ALP) activity, and osteocalcin (OCN) and collagen-I expressions, as well as increased concentration of fasting blood glucose. Moreover, we found that miR-340-5p expression was increased while runt-related transcription factor-2 (RUNX2) was decreased in femurs. Furthermore, the effects of miR-340-5p on osteogenic differentiation (OD) in high glucose (HG)-treated MC3T3-E1 cells were explored. Exposure to OD and HG contributed to elevated miR-340-5p level. Inhibition of miR-340-5p enhanced ALP level, calcium deposition, and OCN, collagen-I and RUNX2 levels. On the contrary, miR-340-5p overexpression reversed these promotional effects. Luciferase assay indicated that RUNX2 may be a target gene of miR-340-5p. Moreover, RUNX2 deficiency decreased miR-340-5p inhibition-induced ALP activity, calcium accumulation and OCN, collagen-I, RUNX2 levels. In short, the above findings revealed that inhibition of miR-340-5p facilitated osteogenic differentiation through regulating RUNX2 in MC3TC-E1 cells, which provided targeted therapeutic strategies for the treatment of DOP.

CAS / MDL: 50-81-7 18883-66-4
研究领域: 分子生物学 · 内分泌学 · 骨生物学 · 糖尿病研究
✓ 已验证 产品关联 DOI PubMed 高影响因子
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MICROCHEMICAL JOURNAL · 2024 · IF 4.9

Dual-mode colorimetric–fluorometric immunoassay for foodborne parasitic infection

Yao Yu,Xiaolei Liu,Ning Xu,Lin Li,Yaming Yang,Mingyuan Liu,Lianjing Zhao,Xue Bai

Immunoassays are the primary method used for detection of foodborne parasitic infection. The nanomaterials used in these assays often present challenges such as complex preparation requirements and low stability during long-term storage and transport. Herein, based on cost-effective K 3 [Fe(CN) 6 ]-mediated in-situ generated CuFe Prussian blue analogues (CuFe-PBA) and quinoxaline derivatives with high fluorescence, a dual-mode immunoassay was proposed for foodborne parasitic infection. This platform is relied on the restoration of K 3 [Fe(CN) 6 ] by ascorbic acid (AA). In addition to this process, K 4 [Fe(CN) 6 ]-triggered the generation of CuFe-PBA, and the enhanced fluorogenic reaction of dehydrogenated AA and o -phenylenediamine (OPD) occur sequentially. Without complex probes preparation, the assay enabled the rapid on-site detection of ALP and foodborne parasitic infection through a colorimetric mode accessible via smartphone and offered enhanced sensitivity through a fluorescent mode. The method was used for the detection of ALP, achieving detection limits of 0.38 U/L and 0.03 U/L for colorimetric and fluorescence modes, respectively. Subsequently, the dual-mode immunoassay was applied to Trichinella spiralis infection. The sensitivity and specificity of both the colorimetric and fluorescence modes were 100 % in 950 pig serum samples. This immunosensor provides a universal analysis tool for the accurate, cost-effective and highly sensitive detection of foodborne parasitic infection.

研究领域: 分析化学 · 食品安全 · 生物技术 · 寄生虫学
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