39 results about "Alp activity" patented technology

The present invention relates to an application of iridoid in the preparation of anti-osteoporosis medicines, which belongs to the medicines technical field. The invention provides an application of iridoid which is shown in a general formula I in the preparation of medicines used for treating and / or preventing osteoporosis. Experiments found that ALP activity in UMR106 cells is taken as guidance for tracing and separating an extract of iridoid possessing general formula I, the extract and each separated component are researched from the aspects of osteoblast propagation and antioxidation activity, the iridoid possessing the general formula I has the effects of substantially promoting alkaline phosphatase activity and osteoblast differentiation, so that the iridoid has anti-osteoporosis effect and can be used for preparing the medicines used for treating or preventing osteoporosis. The component of specnuezhenide and a compound G13 possesses the effects of substantially promoting the alkaline phosphatase activity and osteoblast differentiation, and the specnuezhenide has effects for obviously promoting UMR-106 cell proliferation, a better effect is obtained when the specnuezhenide is used for anti-osteoporosis effect.

The invention discloses a 3D printed sodium alginate-collagen type I-ceramic composite scaffold and a preparation method and application thereof. The composite scaffold is mainly composed of sodium alginate, collagen type I and calcium magnesium silicate according to a mass ratio of (1-6): 2: (0-10) and utilizes calcium chloride as a crosslinking agent, wherein a mass fraction of calcium chloride in the composite scaffold is 1 to 10%. The preparation method comprises dispersing the calcium magnesium silicate powder in deionized water, fully mixing the solution, compounding the solution, collagen type I and sodium alginate to obtain uniform hydrogel, placing the hydrogel in a 3D printer, carrying out crosslinking with calcium chloride and printing a porous scaffold which is the composite scaffold. The composite scaffold can promote chondrocyte proliferation and ALP activity, has excellent biological activity and has an application value in tissue engineering.

The invention provides an alkaline phosphatase activity assay method with high-efficiency signal amplifying function based on in-situ formation of optical active nanometer material mimic enzyme. Hydrolysate products of ALP like catechol or salicylic acid can be combined to the surface of TiO2 nanometer material specially to form charge transfer complex, so that the TiO2 nanometer material can express efficient mimic enzymatic activity under irradiation of visible light. Compared with conventional nanometer mimic enzyme, the optical active mimic enzyme is generated in situ by catalytic reaction of natural enzyme and has excellent biocompatibility without high-concentration H2O2 during use. The optical active nanometer mimic enzyme generated in situ by catalytic reaction of alkaline phosphatase can generate high-efficiency signal amplifying function for activity assay of alkaline phosphatase, so that detection limit of ALP activity is up to 0.01 U / L. The alkaline phosphatase activity assay method has the advantages of simplicity, flexibility and fastness.

The invention discloses an Mg-Ca-Sr-Zn magnesium alloy as well as a preparation method and application thereof. The Mg-Ca-Sr-Zn magnesium alloy consists of Mg, Ca, Sr and Zn in percentage by mass, wherein the content of Ca is 0-5% but not 0, the content of Sr is 0-5% but not 0, the content of Zn is 0-10% but not 0, and the remnants are Mg. Due to cooperation of component design and preparation process, the purposes of conditioning and controlling the mechanical property and the decomposition velocity of medical implants can be achieved, optimal mechanical property and corrosion resistance can be achieved, relatively good biocompatibility can be achieved, the Mg-Ca-Sr-Zn magnesium alloy has the main characteristic of relatively high cell survival rate, relatively high ALP activity and good erythrocyte and hemoglobin compatibility, and the cell survival rate is not remarkably reduced along with the culture time; metal elements essential for human bodies are adopted, no harmful element or element with potential harm is used, and the alloying element Sr can promote osteogenesis and inhibit bone resorption, so that along with decomposition of the Mg-Ca-Sr-Zn magnesium alloy, tissue heal can be promoted while Sr is partially released.

The invention relates to a Beta-TCP porous material and a preparation method thereof. Natural plant fiber or polyurethane sponge is used as a pore creating material. A three-dimensional-net-shaped skeleton green body is prepared by immersing the pore creating material into a slurry made from Beta-TCP which is doped with a trace element Zn. The Beta-TCP porous material is obtained by drying and sintering the three-dimensional-net-shaped skeleton green body. The Beta-TCP material which is doped with the Zn better satisfies the need for material degradation and osseous tissue creeping growth. In the invention, the addition of the trace element Zn into the Beta-TCP porous material improves the ALP activity and promotes the degrading of the Beta-TCP porous material in the case of low porosity, thereby greatly improving the pressive strength and the degrading capability of the Beta-TCP porous material and solving the problem that the mechanical property and the biodegradation property of a Beta-TCP porous biological ceramics embedded material are not matched. The Beta-TCP porous material has good degradation property and good mechanical property, is expectable to improve the defects of bone defect repairing and rebuilding materials, and can be widely and clinically used.

The subject invention relates in part to the surprising and unexpected discovery that insects that are resistant to Bacillus thuringiensis Cry toxins have measurably altered alkaline phosphatase (ALP) activity as compared to insects that are susceptible to Cry toxins. This and other surprising discoveries reported herein have broad implications in areas such as managing and monitoring the development of insect resistance to B.t. toxins. For example, the subject invention provides a simple and fast assay (enzymatic or otherwise) for detecting ALP activity levels and thereby monitoring the development of resistance by insects to crystal protein insect toxins. There was no prior motivation or suggestion to go about resistance monitoring using this simple and easy approach.

The invention discloses an imidacloprid fluorescence immunoassay method based on gold nano-cluster anchored hydroxy cobalt oxide nanosheet, and belongs to the technical field of the biological sensor.A nano composite material is formed by anchoring the gold nano-cluster on a surface of the two-dimensional hydroxyl cobalt oxide nanosheet, and the fluorescence intensity is obviously reduced. The ascorbic acid capable of triggering CoOOH nanosheet decomposition is imported to effectively reverse a quenching effect. Noticeable, the corresponding fluorescence reaction induced by the ascorbic acidis related to the ALP activity of the antibody mark. After the competitive immunoreaction, the antibody of the ALP mark can be combined with the immobilized antigen, the fluorescence change of a detection platform can be adjusted. By utilizing the fluorescence switching of the system, the detection concentration (IC50) on the imidacloprid by the FIA is 1.3ng mL-1, and is 60 times sensitive than the the conventional ELISA (86.4ng mL-1). The fluorescence immunoassay method can realize the high-sensitive detection of the target antigen imidacloprid, a new prospect is opened for the pesticide detection, and an effective policy is opened for the fluorescence immunoassay.

The invention relates to a method for detecting activity of acid phosphatase based on light-controlled enzyme cascade reaction. The method is characterized in that a novel enzyme cascade catalyzing reaction system is established by a photoactivity nanometer material which is produced by natural enzyme through catalyzing reaction in situ, namely the catalyzing product of the acid phosphatase can becombined to the surface of titanium dioxide nanoparticle in situ to form the photoactivity nanometer material; the photoactivity nanometer material is used for activating horse radish peroxidase to catalyze and oxidize the reaction of feature substrate under the radiation of visible light, so as to form the enzyme cascade catalyzing system which can be controlled by light to realize accurate regulating and controlling effects. The novel light-controlled enzyme cascade catalyzing system has the advantages that the high-efficiency triple signal amplifying function is realized, and the acid phosphatase can be simply and conveniently determined at high efficiency and high selectivity; the detection limit of ALP activity can reach 1.0mU / L.

The invention belongs to the technical field of analytical chemistry, and relates to an alkaline phosphatase activity colorimetric detection method based on CeVO4. The alkaline phosphatase activity colorimetric detection method comprises the following steps: respectively adding alkaline phosphatase with different activities and 0.05 mL of 1 mM sodium hexametaphosphate (NaPO3) 6 into a 5 mL centrifuge tube; sequentially adding 0.1 mL of 1mg / mL CeVO4 suspension, 0.1 mL of 5mM TMB and an acetate buffer solution into the mixed solution, wherein the final total volume of the solution is 3mL; afterpassing through the membrane, recording the absorbance at the wavelength of 652 nm by using an ultraviolet-visible absorption spectrophotometer, and drawing an ALP activity-absorbance standard workingcurve; performing calibration; and measuring the absorbance of the sample to be measured, and comparing to obtain the ALP activity. The method is mild in detection process condition, low in detectioncost and simple to operate; the alkaline phosphatase activity is detected through a TMB + CeVO4 system, the detection limit is as low as 0.68 U / L, and the detectable range is as wide as 1-210 U / L; the alkaline phosphatase activity is detected by using a TMB + CeVO4 system, the response to alkaline phosphatase is sensitive, the measurement result of an actual sample is accurate and reliable, and the detection of the alkaline phosphatase activity in human serum is realized.

The invention belongs to the technical field of mimic enzyme and analytical chemistry, and relates to a sulfuration modified CoOx-based alkaline phosphatase activity colorimetric detection method, which comprises respectively adding alkaline phosphatase with different activities and 1 mM HMPi to a 5 mL centrifuge tube, and reacting for 30-60 min; sequentially adding the vulcanization modified CoOxsuspension, TMB, H2O2 and an acetate buffer solution into the mixed solution, uniformly mixing the system, and reacting for 1-30 minutes; recording the absorbance at the wavelength of 652 nm by usingan ultraviolet-visible absorption spectrophotometer after passing through the film; drawing a calibrated ALP activity-absorbance standard working curve according to the absorbance measurement value of the TMBox and the corresponding ALP activity; repeating the above steps on the ALP sample to be detected, and comparing the ALP sample with the standard working curve to obtain the ALP activity. Themethod has the advantages of mild conditions in the detection process, does not need other reagents, realizes convenient and rapid detection of the ALP activity, is low in detection cost, simple in operation, has wide detectable range of 0.8-320 U / L, and realizes detection of the alkaline phosphatase activity in human serum.

The purpose of this study was to identify the periodontal regeneration factors in enamel protein shown to induce cementum- and osteo-promotive activities in vivo. The cementum regeneration (CR), a part of periodontal regeneration, was examined by using experimental cavities prepared in a buccal dehiscence dog model. The CR activity was found in the aggregate consisted of sheath proteins along with a small amount of amelogenins separated from newly formed secretory enamel. The sheath proteins were purified to be homogeneous, and examined for the alkaline phosphatase (ALP) inducing activity of human periodontal ligament (HPDL) cells. Application of 17 kDa sheath protein induced ALP activity in HPDL cells. Peptides synthesized from the sheath protein sequence also showed ALP inducing activity. The 17 kDa sheath protein has cell differentiation activity.

The invention provides a method for quantitatively detecting alkaline phosphatase based on a surface enhanced Raman spectroscopy technology by taking BCIP as a substrate and DMSO as an internal standard substance. Results show that the ALP activity and the intensity ratio (600cm <-1 > / 677cm <-1 >) of the characteristic peak to the internal standard peak have a good linear relationship, and the correlation coefficient is 0.977; the ALP activity in a seawater sample is successfully and quantitatively detected by using the model, and rapid detection of the ALP activity in seawater is realized. Meanwhile, the method can also be applied to activity detection of extracellular enzymes of other microorganisms in seawater, and a solid scientific foundation is laid for in-situ detection of the activity of the extracellular enzymes of the microorganisms in the seawater.

The invention relates to a verification method for stimulating osteogenic differentiation of valvular interstitial cells by inorganic phosphate. The method comprises the following steps: respectivelyperforming osteogenic differentiation culturing on extracted rat valvular interstitial cells by using CON, IP-OIM and OIM and then extracting protein to prepare a protein sample; after ALP staining and alizarin red staining on three kinds of protein samples, evaluating an osteogenic differentiation degree after staining by using ALP activity detection and a relative calcium content determination method, and detecting expression changes of BMP-2, RhoA and Rock-1 through a WB method and an RT-PCR method respectively; with the si-RNA technology, respectively interfering the expression of BMP-2 and RhoA and observing the relationship of mutual changes between the BMP-2 and the RhoA and whether the BMP-2 and the RhoA have the effect of resisting osteogenic differentiation or not after interference; and studying the influence of IP-OIM on SMAD1 / 5 / 9 phosphorylation as well as a mutual relation between two paths of BMP-2 / SMAD1 / 5 / 9 and RhoA / Rock-1. According to the invention, facts that calcification formation of valvular interstitial cells is caused by inorganic phosphate through BMP-2 / smald1 / 5 / 9 and RhoA / Rock-1 signal pathways and the BMP-2 / SMAD1 / 5 / 9 pathway is an upstream pathway of theRhoA / Rock-1 pathway in the valvular interstitial cell osteogenic differentiation process are determined; and a forward regulation effect is realized.

The invention relates to a BMP-2 activator and application of the same in stem cell induced differentiation. Alkaline phosphatases (ALPs), early osteogenesis markers, are mainly distributed in cytomembranes to promote cell calcification, quantitative detection of the ALPs can reflect the differentiation level of osteoblasts, and the higher the activity is, the more obvious differentiation of preosteoblasts into the mature osteoblasts is; the high expression of the ALP activity is an early indicator for osteogenic differentiation maturation, bone formation is enhanced when the ALP activity is improved, and bone matrix mineralization is promoted, so that the ALP activity is a good indicator reflecting the differentiation degree and the functional status of the osteoblasts; to-be-tested compounds 1-3 promote osteogenic differentiation of hBMSCs by activating BMP-2 protein expression in the hBMSCs, and to-be-tested compounds 4 and 5 promote osteogenic differentiation of the hBMSCs by activating VEGF165 protein expression in the hBMSCs.

The invention relates to vascular endothelial growth factor 165 activators and application of the same to stem cells. Alkaline phosphatase (ALP) is an early osteogenesis marker, is mainly distributedin cell membranes and can promote cell calcification. Quantitative detection of ALP can reflect the differentiation level of osteoblasts. The higher the activity of ALP is, the more obvious the differentiation of preosteoblasts into mature osteoblasts is. The high expression of ALP activity is an early indicator for the differentiation and maturation of osteoblasts. When ALP activity is enhanced,bone formation is improved and bone matrix mineralization formation is promoted. Therefore, ALP activity is a good indicator reflecting the differentiation degree and functional status of osteoblasts.To-be-tested compounds 1 to 3 promote the osteogenic differentiation of hBMSCs by activating BMP-2 protein expression in hBMSCs; and to-be-tested compounds 4 and 5 promote the osteogenic differentiation of hBMSCs by activating VEGF-165 protein expression in hBMSCs.

The invention discloses a Mg-Ca-Sr-Zn series magnesium alloy, a preparation method and application thereof. The magnesium alloy is composed of Mg, Ca, Sr and Zn; by weight percentage, in the magnesium alloy, the content of Ca is 0-5%, but not 0, and the content of Sr is 0-5%, but not 0, the content of Zn is 0-10%, but not 0, and the rest is Mg. The present invention regulates the mechanical properties and degradation speed of medical implants through the cooperation of composition design and preparation process, and obtains the best mechanical properties and corrosion resistance; it has better biocompatibility, mainly manifested in higher biocompatibility Cell survival rate, and the cell survival rate does not decrease significantly with the prolongation of culture time, has high ALP activity, good compatibility of red blood cells and hemoglobin; selects essential metal elements for human body, does not contain harmful or potentially harmful elements, of which The alloying element Sr in Mg-Ca-Sr-Zn series magnesium alloy can promote osteogenesis and inhibit bone resorption, so the local release of Sr along with the degradation of Mg‑Ca‑Sr‑Zn series magnesium alloy can promote tissue healing.

The invention belongs to the technical field of medicine, and relates to the application of an iridoid compound in the preparation of anti-osteoporosis drugs. The application of the iridoid compound of general formula I of the present invention in the preparation of medicines for treating and / or preventing osteoporosis. The experiment found that the extract of iridoids with the general formula I was tracked and separated based on the ALP activity in UMR106 cells, and the effects of the extract and the separated components on osteoblast proliferation and antioxidant activity Research has been carried out, showing that the iridoid compounds with the general formula I can significantly promote the activity of alkaline phosphatase and promote the differentiation of osteoblasts, so this type of compound has an anti-osteoporosis effect and can be used to prepare Medicines to treat or prevent osteoporosis. Among them, privetin and compound G13 have the effect of significantly promoting the activity of alkaline phosphatase and promoting the differentiation of osteoblasts, and the effect of privetin can also significantly promote the proliferation of UMR-106 cells, which is used for anti-osteoporosis When the effect is better.

The invention relates to a vascular endothelial growth factor 165 activator and its application in stem cells. Alkaline phosphatase (ALP) is a marker of early osteogenesis, which is mainly distributed in the cell membrane and promotes cell calcification. The quantitative detection of ALP can reflect the differentiation level of osteoblasts. Cell differentiation is more obvious. High expression of ALP activity is an early marker of osteoblast differentiation and maturation. When ALP activity is enhanced, bone formation is enhanced and bone matrix mineralization is promoted. Therefore, ALP activity is a good indicator to reflect the degree of differentiation and functional status of osteoblasts. Test compounds 1-3 promote the osteogenic differentiation of hBMSCs by activating the expression of BMP-2 protein in hBMSCs, and test compounds 4 and 5 promote the osteogenic differentiation of hBMSCs by activating the expression of VEGF-165 protein in hBMSCs.

The present invention relates to a pharmaceutical composition for the prevention or treatment of CFC (cardiofaciocutaneous) syndrome comprising a TGF-β signaling pathway inhibitor or a BMP signaling pathway activator. The treatment of SB-431542 or BMP4 protein was confirmed to increase the ALP activity and bone mineral deposition in the osteoblasts originated from the induced pluripotent stem cells derived from CFC syndrome patients. Thus, the TGF-β signaling pathway inhibitor containing SB-431542 or the BMP signaling pathway activator containing BMP4 protein can be effectively used for the prevention or treatment of CFC syndrome.

The invention discloses a method for analyzing an anti-osteoporosis action mechanism of ginseng-mulberry, and belongs to the technical field of plant drug pair research. The method comprises the following steps: combining in-vitro pharmacodynamics with network pharmacology and a molecular docking technology to research the influence of single ginseng and mulberry and different polar components of the drug pair on the proliferation rate and the testosterone secretion of mouse leydig cells TM3 and the influence of the different polar components of the drug pair on the proliferation rate and the alkaline phosphatase ALP activity of osteoblasts MC3T3-E1; respectively collecting chemical components, components and disease-related targets of ginseng and mulberry through a traditional Chinese medicine component database TCMSP, a GeneCards and an OMIM database, taking an intersection of the components and the disease-related targets, analyzing protein interaction between key targets by applying an STRING database, and carrying out biological function and pathway analysis by utilizing a DAVID database; adopting Cytoscape 3.7.0 to perform composition and network topology structure analysis on a related result; and carrying out molecular docking on potential effective substances and the key targets by using an Autodock software.

The purpose of this study was to identify the periodontal regeneration factors in enamel protein shown to induce cementum- and osteo-promotive activities in vivo. The cementum regeneration (CR), a part of periodontal regeneration, was examined by using experimental cavities prepared in a buccal dehiscence dog model. The CR activity was found in the aggregate consisted of sheath proteins along with a small amount of amelogenins separated from newly formed secretory enamel. The sheath proteins were purified to be homogeneous, and examined for the alkaline phosphatase (ALP) inducing activity of human periodontal ligament (HPDL) cells. Application of 17 kDa sheath protein induced ALP activity in HPDL cells. Peptides synthesized from the sheath protein sequence also showed ALP inducing activity. The 17 kDa sheath protein has cell differentiation activity.

The invention relates to bone morphogenetic protein 2 activators and application of the same to promotion of the osteogenic differentiation of mesenchymal stem cells. Alkaline phosphatase (ALP) is anearly osteogenesis marker, is mainly distributed in cell membranes and can promote cell calcification. Quantitative detection of ALP can reflect the differentiation level of osteoblasts. The higher the activity of ALP is, the more obvious the differentiation of preosteoblasts into mature osteoblasts is. The high expression of ALP activity is an early indicator for the differentiation and maturation of osteoblasts. When ALP activity is enhanced, bone formation is improved and bone matrix mineralization formation is promoted. Therefore, ALP activity is a good indicator reflecting the differentiation degree and functional status of osteoblasts. To-be-tested compounds 1 to 3 promote the osteogenic differentiation of hBMSCs by activating BMP-2 protein expression in hBMSCs; and to-be-tested compounds 4 and 5 promote the osteogenic differentiation of hBMSCs by activating VEGF165 protein expression in hBMSCs.

The invention discloses a 3D printed sodium alginate-collagen type I-ceramic composite scaffold and a preparation method and application thereof. The composite scaffold is mainly composed of sodium alginate, collagen type I and calcium magnesium silicate according to a mass ratio of (1-6): 2: (0-10) and utilizes calcium chloride as a crosslinking agent, wherein a mass fraction of calcium chloride in the composite scaffold is 1 to 10%. The preparation method comprises dispersing the calcium magnesium silicate powder in deionized water, fully mixing the solution, compounding the solution, collagen type I and sodium alginate to obtain uniform hydrogel, placing the hydrogel in a 3D printer, carrying out crosslinking with calcium chloride and printing a porous scaffold which is the composite scaffold. The composite scaffold can promote chondrocyte proliferation and ALP activity, has excellent biological activity and has an application value in tissue engineering.

The purpose of this study was to identify the periodontal regeneration factors in enamel protein shown to induce cementum- and osteo-promotive activities in vivo. The cementum regeneration (CR), a part of periodontal regeneration, was examined by using experimental cavities prepared in a buccal dehiscence dog model. The CR activity was found in the aggregate consisted of sheath proteins along with a small amount of amelogenins separated from newly formed secretory enamel. The sheath proteins were purified to be homogeneous, and examined for the alkaline phosphatase (ALP) inducing activity of human periodontal ligament (HPDL) cells. Application of 17 kDa sheath protein induced ALP activity in HPDL cells. Peptides synthesized from the sheath protein sequence also showed ALP inducing activity. The 17 kDa sheath protein has cell differentiation activity.

The invention discloses an ALP activity detection kit based on a photo-ATRP signal amplification strategy and a use method thereof. The kit comprises the following raw materials: an electrode, MPA, EDC, NHS, O-phosphoethanolamine, BIBB, DMSO, EY, Me6TREN, FMMA and LiClO4. The photo-ATRP reaction is used as a polymerization reaction signal amplification strategy, so that the sensitivity of ALP activity detection is remarkably improved, meanwhile, the use of a transition metal catalyst is avoided, and the method has the advantages of high efficiency, convenience in operation and environmental friendliness. Under the optimal experimental conditions, the linear range of the method for ALP activity detection is 10-150mU / mL, and the limit of detection (LOD) is 2.12 mU / mL, so that the method has good sensitivity. The method provided by the invention has satisfactory selectivity, anti-interference performance, reproducibility and stability, and can obtain good experimental results in clinical serum sample and inhibition rate experiments.

The invention relates to application of a Thiobacillus ferrooxidans BY3 leachate of native copper in preparation of a medicine for promoting differentiation of bone marrow mesenchymal stem cells. According to the invention, the effect of the Thiobacillus ferrooxidans BY3 leachate of native copper in a process of promoting osteogenic differentiation of bone marrow mesenchymal stem cells is studiedby taking bone marrow mesenchymal stem cells of rats as a target object. Results show that the Thiobacillus ferrooxidans BY3 leachate of native copper can enhance the ALP activity of bone marrow mesenchymal stem cells in vitro, promote deposition of calcium salts in bone marrow mesenchymal stem cells, and promote osteogenic differentiation of bone marrow mesenchymal stem cells.

The invention provides a preparation method of myricetin-loaded nano-micelles for reducing ovariectomy-induced bone loss by inhibiting formation of osteoclasts, and belongs to the field of pharmaceutical preparations. The invention discloses a myricetin-loaded nano-micelle, and aims to provide a myricetin-loaded nano-micelle capable of improving the drug loading capacity so as to improve the solubility and oral bioavailability of myricetin and reduce ovariectomy-induced bone loss by inhibiting the formation of osteoclasts. According to the key point of the technical scheme, the preparation method of the myricetin-loaded nano-micelle (hereinafter referred to as myricetin micelle) comprises the step of preparing the myricetin bone targeting micelle from a bone targeting material AL-P (LLA-CL)-PEG-P (LLA-CL) and myricetin. The prepared myricetin micelle has smaller particle size and higher encapsulation efficiency, the in-vitro solubility and in-vivo bioavailability of the myricetin micelle are enhanced, the ALP activity of osteoblasts is remarkably improved, and the treatment effect of resisting castrated rat osteoporosis is enhanced. And preliminary information is provided for clinical application of myricetin and other similar water-insoluble compounds.

The invention belongs to the technical field of simulated enzymes and analytical chemistry, and relates to a colorimetric detection method for alkaline phosphatase activity based on sulfide-modified CoOx, comprising: adding alkaline phosphatase and 1 mM HMPi, reacted for 30-60 min; then added sulfide-modified CoOx suspension, TMB, H 2 o 2 and acetate buffer, mix the system and react for 1 to 30 min; after passing through the membrane, record the absorbance at a wavelength of 652 nm with a UV-visible absorption spectrophotometer; compare the absorbance measurement value of the above TMBox with the corresponding ALP activity Draw the calibrated ALP activity-absorbance standard working curve; repeat the above steps for the ALP sample to be tested, and compare it with the standard working curve to obtain the ALP activity. The detection process of the invention has mild conditions, does not require other reagents, realizes convenient and rapid detection of ALP activity, low detection cost, simple operation, wide detection range of 0.8-320 U / L, and realizes detection of alkaline phosphoric acid in human serum Detection of enzyme activity.

The invention relates to a VEGF-165 activator and its use for promoting stem cell differentiation. Alkaline phosphatase (ALP) is a marker of early osteogenesis, which is mainly distributed in the cell membrane and promotes cell calcification. The quantitative detection of ALP can reflect the differentiation level of osteoblasts. Cell differentiation is more obvious. High expression of ALP activity is an early marker of osteoblast differentiation and maturation. When ALP activity is enhanced, bone formation is enhanced and bone matrix mineralization is promoted. Therefore, ALP activity is a good indicator to reflect the degree of differentiation and functional status of osteoblasts. Test compounds 1-3 promote the osteogenic differentiation of hBMSCs by activating the expression of BMP-2 protein in hBMSCs, and test compounds 4 and 5 promote the osteogenic differentiation of hBMSCs by activating the expression of VEGF-165 protein in hBMSCs.