Detection Of Phosphatase Research Articles

Parthenolide Has Negative Effects on In Vitro Enhanced Osteogenic Phenotypes by Inflammatory Cytokine TNF-α via Inhibiting JNK Signaling.

Nuclear factor kappa B (NF-κB) regulates inflammatory gene expression and represents a likely target for novel disease treatment approaches, including skeletal disorders. Several plant-derived sesquiterpene lactones can inhibit the activation of NF-κB. Parthenolide (PTL) is an abundant sesquiterpene lactone, found in Mexican Indian Asteraceae family plants, with reported anti-inflammatory activity, through the inhibition of a common step in the NF-κB activation pathway. This study examined the effects of PTL on the enhanced, in vitro, osteogenic phenotypes of human periosteum-derived cells (hPDCs), mediated by the inflammatory cytokine tumor necrosis factor (TNF)-α. PTL had no significant effects on hPDC viability or osteoblastic activities, whereas TNF-α had positive effects on the in vitro osteoblastic differentiation of hPDCs. c-Jun N-terminal kinase (JNK) signaling played an important role in the enhanced osteoblastic differentiation of TNF-α-treated hPDCs. Treatment with 1 µM PTL did not affect TNF-α-treated hPDCs; however, 5 and 10 µM PTL treatment decreased the histochemical detection and activity of alkaline phosphatase (ALP), alizarin red-positive mineralization, and the expression of ALP and osteocalcin mRNA. JNK phosphorylation decreased significantly in TNF-α-treated hPDCs pretreated with PTL. These results suggested that PTL exerts negative effects on the increased osteoblastic differentiation of TNF-α-treated hPDCs by inhibiting JNK signaling.

Homogeneous assay based on the pre-reduction and selective cation exchange for detection of multiple targets by atomic spectrometry

In view of the high sensitivity and good selectivity, chemical vapor generation atomic spectrometry (CVG-AS) and inductively coupled plasma mass spectrometer (ICP-MS), especially low-cost atomic fluorescence spectrometry (AFS) have been widely used in bioassay. However, the existing AS method is mostly based on heterogeneous strategies, and can't detect multiple targets in one system. In this study, we present the discovery and mechanism study of a phenomenon of Hg2+ pre-reduction that the concentration of Hg2+ decreased when it was mixed with the reductants (ascorbic acid (AA), SnCl2, or NaBH4/KBH4) over long-time reaction (hours) by CVG-AFS and ICP-MS. A homogeneous Cu2+ assay method was developed based on the competition reaction of Cu2+ and Hg2+ for consuming AA, and its application in the detection of pyrophosphate (PPi) and alkaline phosphatase (ALP) was investigated based on the PPi complexation with Cu2+, and ALP hydrolyzation of PPi using CVG-AFS as a representative detector. Subsequently, in order to further verify the applicability of the system, cation exchange reaction (CER) was utilized here based on the selectively recognize Ag+ and C–Ag+-C by CuS nanoparticles (NPs). As the exchanged Cu2+ from CuS NPs can be sensitively and selectively detected via above-mentioned Cu2+ assay method, this strategy can be extended for the Ag+, DNA and prostate specific antigen (PSA) detection based on base complementary pairing and the specific recognition of aptamer. Under the optimal experimental conditions, the system showed high sensitivity for the detection of Cu2+, PPi, ALP, Ag+, DNA, and PSA, with limit of detections (LODs) of 0.12 nmol L−1, 25 μmol L−1, 0.025 U/L, 0.2 nmol L−1, 0.05 nmol L−1, and 0.03 ng/mL, respectively. The method was successfully used to determination Cu2+, ALP, and PSA in human serums, showing similar results with those of ICP-MS and kits methods.

One-pot cascade catalysis at neutral pH driven by CuO tandem nanozyme for ascorbic acid and alkaline phosphatase detection

During the past decades, most reported nanozymes have been confined to limited species of enzyme-like activities and only few examples were reported to mimic tandem enzyme-like activities. To fill gaps in expanding the species and improving catalytic efficiency of nanozymes, exploring nanozymes as tandem nanozymes has turned out to be an effective method. Herein, we have presented the cupric oxide nanoparticles (CuO NPs) as tandem nanozymes, which is expected to simultaneously catalyze the cascade reaction coupling ascorbate oxidase with peroxidase like activities at temperature (45 °C) and neutral pH (pH = 7). This one-pot cascade reaction system included the oxidation of ascorbic acid (AA) to yield H2O2 and terephthalic acid (TA) oxidation reaction mediated by H2O2 to generate a fluorescence product (λex = 315 nm, λem =422 nm). More significantly, fluorescent sensors are respectively fabricated for AA, alkaline phosphatase (ALP) and l-phenylalanine (ALP inhibitor) detection coupling the catalysis of CuO tandem nanozymes with ALP enzymatic reaction. Base on these findings, this work shows high selectivity/sensitivity and low limit of detection (2.92 × 10−8 M for AA, 0.058 U/L for ALP) properly due to an in-situ reaction. CuO tandem nanozymes expand the species of nanozymes and these CuO tandem nanozymes based fluorescent sensors can be applied for one-pot nonenzymatic biomolecular sensing in clinical diagnostics, biological or pharmaceutical analysis.

Generation of induced pluripotent stem cells from large domestic animals

BackgroundInduced pluripotent stem cells (iPSCs) have enormous potential in developmental biology studies and in cellular therapies. Although extensively studied and characterized in human and murine models, iPSCs from animals other than mice lack reproducible results.MethodsHerein, we describe the generation of robust iPSCs from equine and bovine cells through lentiviral transduction of murine or human transcription factors Oct4, Sox2, Klf4, and c-Myc and from human and murine cells using similar protocols, even when different supplementations were used. The iPSCs were analyzed regarding morphology, gene and protein expression of pluripotency factors, alkaline phosphatase detection, and spontaneous and induced differentiation.ResultsAlthough embryonic-derived stem cells are yet not well characterized in domestic animals, generation of iPS cells from these species is possible through similar protocols used for mouse or human cells, enabling the use of pluripotent cells from large animals for basic or applied purposes. Herein, we also infer that bovine iPS (biPSCs) exhibit similarity to mouse iPSCs (miPSCs), whereas equine iPSs (eiPSCs) to human (hiPSCs).ConclusionsThe generation of reproducible protocols in different animal species will provide an informative tool for producing in vitro autologous pluripotent cells from domestic animals. These cells will create new opportunities in animal breeding through transgenic technology and will support a new era of translational medicine with large animal models.

Facile synthesis of magnetic hierarchical flower-like Co3O4 spheres: Mechanism, excellent tetra-enzyme mimics and their colorimetric biosensing applications

Magnetic hierarchical flower-like Co3O4 spheres (Co3O4 nanoflowers) were facilely prepared via one-step surfactant-free and template-free wet chemical route at room temperature. The formation mechanism of Co3O4 nanoflowers was explored. The as-prepared Co3O4 nanoflowers exhibited excellent tetra-enzyme mimetic activities, including oxidase-like, peroxidase-like, catalase-like and superoxide dismutase (SOD)-like activity. The catalytic mechanism of the Co3O4 nanoflowers was studied in detail. The oxidase-like catalytic activity of Co3O4 nanoflowers was derived from the inherent oxygen vacancies of Co3O4, while the peroxidase-like catalytic activity originated from the •OH radical generated by hydrogen peroxide (H2O2). Only one specific enzyme mimics reaction with Co3O4 nanoflowers can be obtained by inhibiting specifically other nanozymes via varying pH, adding appropriate scavengers or selecting its specific substrate. Further, the steady-state kinetic and catalytic performance of the oxidase-, peroxidase- and catalase mimics of Co3O4 nanoflowers were studied. Based on the oxidase-like and peroxidase-like activities of Co3O4 nanoflowers, bifunctional colorimetric sensing platforms were constructed for the sensitive detection of acid phosphatase (ACP) with the linear range of 0.1–25 U L-1 and H2O2 with the linear range of 4–400 μM. Further, it is capable of detecting ACP in serum samples and H2O2 in water samples, respectively. Thus, the tetra-enzyme mimetic Co3O4 nanoflowers show broad prospects in the fields of biosensors, tumor therapy, environmental monitoring and biocatalysis.

An Enzyme-Induced Metallization-Based Electrochemical Signal Amplification Strategy for Ultrahigh Sensitive Alkaline Phosphatase Detection at Attomolar Concentrations

A highly sensitive enzyme-induced metallization-based electrochemical signal amplification strategy was developed based on the integration of the enzyme-induced metallization reaction, Au nanoparticles (AuNPs)-induced silver deposition, chemically modified electrode and highly sensitive stripping voltammetry detection. The presence of alkaline phosphatase (ALP) catalyzed ascorbic acid 2-phosphate into ascorbic acid, which reduced Ag+ to Ag0 on the surface of the AuNPs/multi-walled carbon nanotubes/polyethyleneimine/glassy carbon electrode (AuNPs/MWNTs/PEI/GCE). As a result, the enzyme-generated product was accumulated on the surface of the AuNPs/MWNTs/PEI/GCE by means of silver deposition via this signal amplification strategy, which enhanced the detection signal dramatically. Amounts as low as 1 × 10–8 U/mL (corresponding to 10 aM) ALP can be detected using this strategy, which is about 4−7 orders of magnitude more sensitive than with other reported methods for ALP determination. In addition, this strategy can be applied to ALP detection in real complex samples, which shows great potential in the early diagnosis of diseases.

Label-Free Colorimetric Detection of Acid Phosphatase and Screening of Its Inhibitors Based on Biomimetic Oxidase Activity of MnO2 Nanosheets.

In this research, we attempted to develop a sensitive colorimetric sensing strategy for the detection of acid phosphatase (ACP) based on MnO2 nanosheets and explored its applications in screening and evaluating inhibitors of ACP. The MnO2 nanosheets exhibit intrinsic biomimetic oxidase activity, which can catalyze the oxidation of the colorless 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonate) diammonium salt (ABTS) into green oxidized ABTS (oxABTS). Upon the introduction of ACP, l-ascorbic acid-2-phosphate can be dephosphorylated to ascorbic acid, which arouses the disintegration of MnO2 nanosheets into Mn2+ ions. This disintegration weakens the enzyme mimicking activity of the MnO2 nanosheets, leading to the impediment of the oxidation of ABTS. Conversely, in the absence of ACP, the ABTS is rapidly oxidized by MnO2, leading to a significant colorimetric signal change. The absorbance difference at 420 nm displayed a linear relationship with the concentration of ACP ranging from 0.075 to 0.45 mU·mL-1, generating a detection limit of 0.046 mU·mL-1. In the inhibition assays, this sensing platform provided simple detection for parathion-methyl (PM), a representative inhibitor of ACP. The facile evaluation of the inhibitory effect of PM, including its IC50 toward ACP, was also realized.

Inhibition of acid phosphatase detection in mixed semen-blood training samples by anti-coagulants present in blood collection tubes

Acid phosphatase detection using fast blue or fast black reagent is routinely performed at the RCMP National Forensic Laboratory Services as a presumptive test for semen. In training exercises, samples consisting of semen mixed with blood were reported to yield negative results with the fast black reagent. To determine the cause of the inhibition, mixed semen-blood stains were prepared using different blood sources, anti-coagulants, and layering techniques, before being tested with the fast blue or the fast black reagent. The anti-coagulant sodium fluoride (NaF) was shown to cause false negative results with the fast black reagent and to impair detection when using the fast blue reagent while no inhibition was observed when dipotassium EDTA (K2 EDTA) was used as anti-coagulant. Four different toothpastes containing fluoride were also tested for potential inhibition of semen detection. Inhibition was observed when semen was mixed with three of the undiluted toothpastes but not with diluted toothpastes (1 in 20 in water). These findings highlight the importance of using appropriate blood sources when preparing mixed semen-blood stains for the purpose of training and proficiency testing in a forensic laboratory. However, forensic casework samples are not expected to contain NaF at the inhibiting levels.

PTEN/PI3K/VEGF signaling pathway involved in the protective effect of xanthine oxidase inhibitor febuxostat against endometrial hyperplasia in rats.

Endometrial hyperplasia (EH) is a medical condition that affects many females as it increases their uterine carcinogenic potential. EH results from entangling hormonal imbalance and inflammatory response. The study examined the role of a xanthine oxidase inhibitor, febuxostat, in a rat model of EH. Adult female Wistar albino rats were subjected to estradiol valerate (EV) 2 mg/kg for 10 days to induce EH. Another group was treated concomitantly with febuxostat 10 mg/kg for the same period. The uterine malondialdehyde, reduced glutathione (GSH), and superoxide dismutase (SOD) were assessed by chemical methods. Gene expressions of phosphatidylinositol-3-kinase (PI3K), Akt, and hypoxia-inducible factor 1 alpha were assessed by the quantitative real-time polymerase chain reaction. Moreover, the vascular endothelial growth factor (VEGF) and interleukin-6 (IL-6) were measured by enzyme-linked immunosorbent assay. Histopathology and immunohistochemical techniques were used for the detection of phosphatase and tensin homolog (PTEN). The results revealed that EV administration induced complex EH with focal atypia and loss of PTEN expression by the histological examination. Uteri of the EV group showed a significant drop in GSH content and SOD activity and rise in the expressions of PI3K, Akt, VEGF, and IL-6. Febuxostat administration significantly improved the uterine GSH and SOD levels. It decreased the expressions of PI3K, Akt, VEGF, and IL-6. The endometrium showed a regression of the proliferative epithelium with the restoration of PTEN expression and the absence of the atypical features. In conclusion, febuxostat protected the endometrium against estrogen-induced EH and may be beneficial in the management along with the hormonal therapy.

Synthesis of water soluble CuGaS2/ZnS quantum dots for ultrasensitive fluorescent detection of alkaline phosphatase based on inner filter effect

Developing monitoring technique for alkaline phosphatase (ALP) is crucial due to the important role it plays in living cells. Here, a kind of biocompatible glutathione-modified CuGaS2/ZnS quantum dots (GSH-CGS/ZnS QDs) was used as a fluorescent substance and then fabricated “turn-off” fluorescent biosensor for detection of ALP by help of inner filter effect (IFE). Firstly, we prepared CuGaS2/ZnS (CGS/ZnS) QDs using solvothermal method and explored the efficient ligand (GSH) exchanges strategy for transferring oil-soluble CGS/ZnS QDs to aqueous phase. More importantly, we also explored the potential biological applications of the nanohybrid QDs. The obtained GSH-CGS/ZnS QDs emitted strong yellow fluorescence with the maximum excitation (400 nm) and emission (601 nm). Then, GSH-CGS/ZnS QDs were mixed with p-nitrophenylphosphate (PNPP) and ALP. PNPP could be hydrolyzed to p-nitrophenol (PNP) by help of catalysis of ALP, and the excitation spectrum of the GSH-CGS/ZnS QDs overlapped well with the absorption spectrum of PNP, so the fluorescence of GSH-CGS/ZnS QDs was initially quenched via the so-called “IFE”. Finally, a novel “turn-off” biosensor for sensitive detection of ALP in the range of 0.05–10 U L −1(R2 = 0.98) with a detection limit of 0.01 U L−1 was successfully obtained. Results indicated that I-III-VI2 nanocrystals have great potential for their promising biomedical application.

Physical Properties, Antimicrobial Activity and In Vivo Tissue Response to Apexit Plus

We investigated the physical properties, antimicrobial activity, and tissue reaction to Apexit Plus in comparison to Sealapex. Flow, radiopacity, setting time, and solubility were evaluated in each material. The antimicrobial activity against Enterococcus faecalis was performed. Polyethylene tubes containing Apexit Plus or Sealapex, and without material (control group) were implanted into the subcutaneous tissue of rats. At 7, 15, 30, and 60 days of implantation, the specimens were paraffin-embedded and the number of inflammatory cells (ICs) and the amount of birefringent collagen (BC) were quantified. The von Kossa reaction followed by immunohistochemistry for detection of alkaline phosphatase (ALP) was also performed. Statistical analysis was performed with ANOVA and Tukey test (p ≤ 0.05). The flow value of Apexit Plus was greater than Sealapex, whereas the radiopacity (3.44 mm Al) was lower than Sealapex (6.82 mm Al). Apexit Plus showed lower solubility and shorter initial and final setting (p < 0.0001), whereas the antimicrobial activity was significantly greater than Sealapex. Although the number of ICs was higher in Apexit Plus (p = 0.0009) at 7 days, no significant difference was detected between Apexit Plus and Sealapex at 15, 30, and 60 days. All groups showed higher values for BC in the capsules over time. ALP-immunolabelled cells were observed, mainly around von Kossa-positive structures, either in the capsules of Apexit Plus or Sealapex. Therefore, our results revealed that Apexit Plus exhibited a greater effectiveness against Enterococcus faecalis and better physical properties than Sealapex, except for the radiopacity. In vivo findings indicate that Apexit Plus is biocompatible and presents potential bioactivity in the subcutaneous tissue.

A portable photoacoustic device for facile and sensitive detection of serum alkaline phosphatase activity

It is still a high challenge to develop a simple, sensitive and portable approach for bioassay in strong scattering medium. Herein, a photoacoustic (PA) device is developed for the detection of alkaline phosphatase (ALP) in serum with silver nanoparticles (AgNPs) as signal probe, without any requirements for expensive equipment, professional operation and pre-processing of real samples. ALP as an important disease marker could catalyze the breakdown of sodium L-ascorbyl-2-phosphate (AAP) into ascorbic acid (AA), thereby reducing Ag+ to AgNPs. AgNPs could generate strong PA signal under the irradiation of modulated 638-nm laser due to their localized plasmon resonance, and detected by the self-made portable PA device. Under the optimized experimental conditions, the present PA device exhibits excellent photostability and reproducibility with the relative standard deviation (RSD) of 2.2% at the concentration of 25 U L−1 ALP. Linear calibration graph is obtained within 5–70 U L−1 for ALP, along with a detection limit of 1.1 U L−1. This portable PA device is applied to detect ALP in serum samples, providing satisfactory spiking recoveries and competitive analytical performances with the current techniques. The PA-based analytical strategy obviously opens up a new avenue to the detection of disease-correlated biomarker in practice.

Enzyme-responsive Fluorescent Ionic Liquid.

Herein, we describe the development of a novel material, "enzyme-responsive fluorescent ionic liquid", which enabled a highly sensitive detection of alkaline phosphatase (ALP). We prepared a plasticized poly(vinyl chloride) (PVC) membrane using this new material as a plasticizer and quantified ALP in aqueous solutions. Preliminary results suggested that the PVC membrane responded to ALP at an interface between the membrane and the sample solution with anion extraction to maintain electroneutrality in the membrane phase. The developed PVC membrane showed an approximately six-times higher sensitivity than the conventional membrane, thereby demonstrating highly sensitive ALP detection. These results suggested the potential applicability of the proposed membrane for highly sensitive protein detection by using ALP-labeled antibodies.

O-Phenylenediamine/gold nanocluster-based nanoplatform for ratiometric fluorescence detection of alkaline phosphatase activity

This study demonstrates a novel and convenient ratiometric fluorescent method for the detection of alkaline phosphatase (ALP) activity. Amino-functionalized mesoporous silica nanoparticle-gold nanoclusters (MSN-AuNCs) nanocomposites were integrated with o-phenylenediamine (OPD) to form a ratiometric fluorescence nanoplatform. The presence of ALP induced the generation of quinoxaline (QX) derivative which called 3-(dihydroxyethyl)furo[3,4-b]quinoxaline-1-one (DFQ) with strong fluorescence emission at 450 nm, while the orange-red fluorescence of MSN-AuNCs at 580 nm was slightly quenched. Meanwhile, an obvious fluorescence color change from orange-red to purple and finally to blue can be observed by naked eyes with the increasing of ALP concentration. Therefore, employing the fluorescence emission of DFQ at 450 nm as the reporter signal and the fluorescence emission of MSN-AuNCs at 580 nm as a reference signal, a sensitive ratiometric detection method for ALP was developed. Quantitative detection of ALP activity in the linear range from 0.2 to 80 U/L with a detection limit of 0.1 U/L can be realized in this way, which endows the assay with high sensitivity enough for practical detection of ALP in human serum samples.

Dual-emission carbon dots for ratiometric detection of Fe3+ ions and acid phosphatase.

We have developed a simple and convenient route to prepare fluorescent carbon dots with dual emission peaks respectively at 470 and 570nm. The prepared dual-emission carbon dots can be used for ratiometric detection of Fe3+ ions in the range from 0 to 50μmol·L-1 with 0.8μmol·L-1 detection limit based on the fluorescence quenching at 570nm. The quenched fluorescence induced by Fe3+ ions could be recovered by pyrophosphate. We further used the carbon dots-Fe3+ ions-pyrophosphate mixed system for ratiometric detection of acid phosphatase in the range from 0.08 to 6.75μg·mL-1 with 0.01μg·mL-1 detection limit.

Multi-emitting fluorescence sensor of MnO2–OPD–QD for the multiplex and visual detection of ascorbic acid and alkaline phosphatase

A blue-, yellow-, and red-emitting fluorescence sensor of MnO2–OPD–QD for the multiplex and visual detection of ascorbic acid and alkaline phosphatase.

Human serum albumin templated MnO2 nanosheets as an efficient biomimetic oxidase for biomolecule sensing.

Herein, we have proposed a colorimetric biosensor for detection of acid phosphatase based on human serum albumin (HSA) templated MnO2 nanosheets (HSA-MnO2 NSs). HSA-MnO2 NSs as an efficient biomimetic oxidase could catalyze the oxidization of 3,3',5,5'-tetramethylbenzidine (TMB) to the coloured oxidation product (oxTMB). Acid phosphatase (ACP) could hydrolyze l-ascorbic acid-2-phosphate (AAP) to produce ascorbic acid, and ascorbic acid could lead to the decomposition of MnO2 NSs to Mn2+ ions, inhibiting the production of oxTMB. On the basis of this, we have demonstrated a novel colorimetric approach for the detection of acid phosphatase with the linear range from 50 μU mL-1 to 1500 μU mL-1 and a detection limit of 40 μU mL-1. The MnO2 NS-based colorimetric method has been successfully used to determine the content of acid phosphatase in real samples with satisfactory results.

A ratiometric fluorescence sensor based on enzymatically activatable micellization of TPE derivatives for quantitative detection of alkaline phosphatase activity in serum.

A novel ratiometric fluorescence assay via enzymatically activatable micellization in aqueous solution was devised for quantitative detection of alkaline phosphatase (ALP) activity. We demonstrated that the dephosphorylation of the water-soluble, phosphate-functionalized, fluorophore monomer P-TPE-TG, induced by an enzymatic reaction of ALP, leads to micelle formation in aqueous solution because its water-soluble functionality is reduced. The dephosphorylation-induced micellization of P-TPE-TG exhibited a ratiometric sensing response for various ALP concentrations (10–200 mU mL−1) and provided a suitable sensing platform for naked eye detection with increased fluorescence quantum yield (Φ = 3.2%), even compared to a typical TPE-based sensor (Φ = 1.0%), where ALP can be sensed with a detection limit of 0.034 mU mL−1. In addition, P-TPE-TG displayed excellent sensing performance at concentrations from 0 to 50 mU mL−1 in diluted human serum (10%), which offers the capability to exploit ratiometric responses for bioactive substances under practical conditions.

Dephosphorylation-directed tricyclic DNA amplification cascades for sensitive detection of protein tyrosine phosphatase.

We develop a new fluorescence method for the sensitive detection of protein tyrosine phosphatase 1B (PTP1B) based on dephosphorylation-directed tricyclic DNA amplification cascades. This method exhibits good specificity and high sensitivity with a detection limit of 0.24 pM. Moreover, it can be applied for kinetics analysis, inhibitor screening, and the accurate detection of PTP1B in a variety of cancer cells.

Phosphate-responsive 2D-metal-organic-framework-nanozymes for colorimetric detection of alkaline phosphatase.

In this study, a simple colorimetric method with a tunable dynamic range for alkaline phosphatase (ALP) activity assay was developed by using a peroxidase-mimicking two-dimensional-metal-organic-framework (2D-MOF). Phosphates including pyrophosphate (PPi), ATP, and ADP inhibited the peroxidase-like activity of the 2D-MOF, while their hydrolytic product, phosphate (Pi), exhibited no such inhibitory ability. Therefore, by integrating with the ALP catalyzed hydrolytic reaction, a colorimetric method was developed for ALP activity quantification. Furthermore, by using different phosphates as inhibitors for the peroxidase-mimicking 2D-MOF, the dynamic range of this colorimetric method could be efficiently modulated. Three linear ranges of 2.5-20 U L-1, 5-60 U L-1, and 50-200 U L-1 could be obtained by using PPi, ATP, and ADP as inhibitors, respectively. Other proteins exhibited negligible interference, demonstrating that the proposed sensing method possessed excellent selectivity for ALP. Moreover, the developed assay was applied to evaluate ALP inhibitors and construct logic gates. This work not only provides a promising method for ALP detection but also represents a major step towards nanozyme bioanalysis.