55 results about "Na k atpase activity" patented technology

The invention relates to a preparation method of a dental implant and a composite surface thereof. In the dental implant, medical titanium or titanium alloy is used as a substrate, and porous morphology is prepared on the surface while polydopamine and hydroxyapatite are deposited to obtain a dental implant composite surface with good bioactivity. In the invention, a uniform multi-layer pore oxidation film is quickly formed on the surface of the titanium substrate to obtain nano porous morphology; and by enabling a thin layer of protein analogue (polydopamine) to tightly adhere to the surface, the surface hydrophilicity and the corrosion resistance of the dental implant can be improved. Finally, the hydroxyapatite deposition is induced on the surface to obtain a three-layer composite surface; moreover, the nano porous composite surface can effectively reinforce the adhesion and spreading of osteoblast on the implant surface as well as the alkaline phosphatase activity, early bone integration of the implant is accelerated, and firm combination of the material and the organism interface is realized so as to improve the stability and success rate of the implant.

The invention relates to a fluorescence biosensor, and a preparation method and a purpose thereof. On the basis of a two-dimensional manganese dioxide nanometer sheet as a fluorescence sensing platform, morpholineethanesulfonic acid is used for reducing potassium permanganate for synthesizing the manganese dioxide nanometer sheets; the manganese dioxide nanometer sheets adsorb FAM-ssDNA and quench FAM fluorescence; under the effect of alkaline phosphatase, vitamin C magnesium L-ascorbate-2-phosphate is hydrolyzed to generate ascorbic acid; the two-dimensional manganese dioxide nanometer sheets are reduced into manganese ions by the ascorbic acid; FAM-ssDNA is released; the fluorescence is recovered; the fluorescence sensor using the manganese dioxide nanometer sheets and the alkaline phosphatase to regulate the fluorescence intensity is built and is used for detecting the activity of the alkaline phosphatase. Compared with the prior art, the fluorescence biosensor has the characteristics of simplicity, high speed, low background signal, high sensitivity, low detection limit and good selectivity by using the change of the two-dimensional manganese dioxide nanometer sheets on the fluorescence signal, i.e., the off-on mode for detecting the alkaline phosphatase.

The invention discloses a method for measuring alkaline phosphatase activity. According to the method, L-ascorbic acid-2-phosphate is taken as a catalytic substrate of alkaline phosphatase, the substrate is hydrolyzed to form L-ascorbic acid under the catalysis of alkaline phosphatase, L-ascorbic acid can reduce bivalent copper into monovalent copper, a bright yellow bivalent copper-bathophenanthroline compound is converted into deep green monovalent copper-bathophenanthroline compound, the higher the alkaline phosphatase activity is, the more obviously the color of a detection liquid changes, and accordingly, semiquantitative colorimetric detection of the alkaline phosphatase activity is realized or quantitative detection is realized through an ultraviolet-visible spectrophotometer. By means of the method, alkaline phosphatase with limit of detection being 3.05 mU*mL<-1> can be detected. Compared with conventional methods, the method has high sensitivity and anti-interference capability and good repeatability, and is simple to operate, short in detection time and low in cost, and a simple, convenient, low-cost, high-resolution and sensitive colorimetric analysis method with stable performance is provided for detection of the alkaline phosphatase activity in a clinic sample.

A neuroprotective pharmacological method in cerebral ischemic insult comprising administration of L- or DL-threo-DOPS or a pharmaceutically acceptable acid addition salt thereof to a patient. Threo-DOPS directly acts on neurons to exert an effect to protect against neuronal death due to brain ischemia, an antilethal effect, and an anti-edema effect due to excessive polarization of neuronal membrane potential caused by an increase in Na-K-ATPase activity. renaline.

The invention belongs to the field of nano-particles, and particularly relates to a method for detecting alkaline phosphatase activity by molybdenum disulfide quantum dot inner-filter effect fluorescence. The method includes the steps: firstly, preparing fluorescent molybdenum disulfide quantum dots: taking molybdenum disulfide powder as a raw material, taking ethyl alcohol/water as a solvent, taking inorganic base as a stripping auxiliary and synthesizing the molybdenum disulfide quantum dots by an ultrasonic method; secondly, adding alkaline phosphatase solution into 4-nitrophenyl phosphatedisodium salt (PNNP) and magnesium sulfate alkaline buffer solution, and performing co-incubation; thirdly, adding the alkaline buffer solution into molybdenum disulfide quantum dot solution and performing further co-incubation; fourthly, building a standard work curve for detecting alkaline phosphatase; fifthly, acquiring the activity of the alkaline phosphatase in the solution to be detected according to a standard curve equation. The fluorescent molybdenum disulfide quantum dots have the advantages of stable performance, simple synthesis conditions, low price and the like, and are simple indetection process and high in selectivity and sensitivity when being used for detecting the alkaline phosphatase.

The invention discloses a compound shown as a formula (i) and a preparing method thereof. The compound can be adopted as a fluorescence probe and applied for an alkaline phosphatase activity fluorescence detection method. By utilization of a fluorescence resonance energy transfer theory, and by adopting a tetraphenylethylene derivative and 5(6)-carboxyl fluoresce as a donor and a receptor for an FRET system, an "OFF-ON"FRET probe adopting a phosphate group as a recognition group is designed and synthesized. Equipment needed for preparation of the fluorescence probe is simple. Operation processes are simple. The sources of raw materials are commercialized and easily available. The probe has high specificity for the alkaline phosphatase. When the fluorescence probe is applied for alkaline phosphatase activity fluorescence detection, the accuracy and the sensitivity are largely improved than those of basic methods used in the prior art for detection the alkaline phosphatase activity.

The invention discloses a simple and low cost method for detecting the activity of alkaline phosphatase. According to the method, after the alkaline phosphatase analyte in a sample solution catalyzes the pyrophosphoric acid hydrolysis, Cu2+ is not complexed with pyrophosphoric acid and thus can high efficiently inhibit the activity of glucoamylase, and starch with low water solubility cannot be hydrolyzed by f glucoamylase. Starch in the reaction solution is dropwise added on a paper micro-fluidic analysis device, the corresponding paper carries out wettability alteration from hydrophilic property to hydrophobic property, the volume of a detection reagent solution that can flow through the paper is reduced, and finally the flow length of the detection reagent solution in the paper device is shortened. The flow length of the detection reagent solution in the paper device and the activity of the alkaline phosphatase in the sample are in a inversely proportional relationship, and thus the quantitative detection of alkaline phosphatase activity can be realized. The method is simple and easy to perform, even an untrained operator can carry out experiments, the quantitative signals can be read without using any instrument, the analysis cost is largely reduced, and the method has a wide application prospect.

The invention provides an electrochemical method used for measuring activity of alkaline phosphatase and an electrochemical sensor used in the method. The method comprises the following steps: preparing an electrode modified by cysteine and copper ions; drawing a standard curve between the enzyme activity concentration of alkaline phosphatase and relative current intensity under a pyrophosphate ion substrate with specific concentration; changing the alkaline phosphatase solution with known enzyme activity concentration into a to-be-measured sample containing alkaline phosphatase, performing cyclic voltammetry scanning to obtain the relative current intensity, and calculating to obtain the activity concentration of the alkaline phosphatase in the to-be-measured sample according to the standard curve. According to the method disclosed by the invention, a reversible competitive coordination principle of the cysteine, pyrophosphate ions and copper ions is utilized, the electrochemical sensor used for measuring the activity of alkaline phosphatase is prepared, and the activity of alkaline phosphatase can be rapidly, simply and accurately measured by utilizing the electrochemical sensor.

The invention discloses a method for simply, conveniently and quickly determining the activity of alkaline phosphatase. The method comprises the following steps: obtaining 5'-end phosphorylated double-stranded linear DNA (deoxyribonucleic acid) of one chain, then reacting in a reaction system in the presence of alkaline phosphatase by taking the double-stranded linear DNA as a substrate, reacting a reactant of the 5'-end phosphorylated double-stranded linear DNA obtained by reaction in the presence of sufficient gamma exonuclease, determining the relative amount of single-stranded DNA or double-stranded DNA in the reaction system after reaction, and deducing the degree of activity of alkaline phosphatase according to the detection result. Compared with the prior art, the method is simple and fast to operate, can realize high-throughput and automated activity determination, further has high sensitivity and can detect 10mU of alkaline phosphatase.

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 a method for screening a pluripotent cell and a special culture medium thereof. A pluripotent cell screening culture medium provided by the invention comprises PD0325901 shown in formula (I) and CHIR99021 shown in formula (II). The method for screening a pluripotent cell provided by the invention comprises the step of using the screening culture medium to culture a cell A, thus obtaining the pluripotent cell; and the cell A is a cell obtained by inducing an inducible factor encoding gene in a differentiation cell. The pluripotent cell obtained by applying the method of the invention has the protuberant cloned shape appearance of mESC; the symbol gene and the surface protein of an express embryonic stem cell have the activity of surface alkaline phosphatase and the capacity of automatically differentiating to form an Embryoid body as the embryonic stem cell;, and the pluripotent cell can be further differentiated into the cell types of an inner embryonic layer, a middle embryonic layer and an outer embryonic layer. The invention has important significance for the research field of the pluripotent cell.

The invention provides a method for detecting alkaline phosphatase activities. The method comprises the following steps: A1) uniformly mixing tri(hydroxymethyl) aminomethane and pyrophosphate, and then dividing into a plurality of reaction systems, respectively adding alkaline phosphatase with different concentrations into all the reaction systems, incubating at 37 DEG C, adding an acetate buffersolution, CeO2 nanoparticles and 3,3',5,5'-tetramethyl benzidine into each reaction system in turn, incubating at 22-47 DEG C, and then respectively measuring ultraviolet-visible absorption spectrum,thus obtaining a relation equation between alkaline phosphatase activities and Delta A; A2) measuring the ultraviolet-visible absorption spectrum for the to-be-detected alkaline phosphatase accordingto the operation in the step A1, thereby acquiring Delta A, substituting the Delta A into the relation equation of alkaline phosphatase activities and Delta A acquired in the step A1, and calculatingthe activities of the to-be-detected alkaline phosphatase. The method has the beneficial effects: 1) the CeO2 nanoparticles utilized by the invention are excellently dispersed in an aqueous solution and 2) the method provided by the invention is short in detecting time.

The invention relates to novel uses of rhodiola effective ingredient in prevention and cure for osteoporosis. As experiment demonstrates: the compound can obviously upgrade osteotropic protein bmp-2 and mRNA expression in an osteoporosis-resisting medicine screening model, obviously facilitates osteoblast proliferation of human, alkaline phosphatase activity and collagen synthesis and increase osteogenous bone formation, thereby possessing prospect of medicine for prevention and cure for osteoporosis.

The invention discloses a method for detecting and evaluating in-vitro cell morphology and osteogenic function of the surface of a bone substitute implant, and belongs to the technical field of biomaterial detection. The method is characterized by comprising the following steps: taking the bone substitute implant to be detected and evaluated, firstly, observing the cell morphology, cytoskeleton morphology and skeleton protein distribution condition, then detecting cell activity, cell adhesion on the surface of the material and alkaline phosphatase activity in cells, then detecting collagen secretion and the external parts of the cells, and remembering the mineralization degree, and finally, determining the cell morphology as well as the expression condition of framework related genes and osteogenesis related genes. According to the method, biological reference is better provided for optimal design of the surface of the bone substitute implant, and on the basis of the prior art, lots of detecting methods are added, and part detecting methods are improved, so that the method is comprehensive and systematical, and passes the cytology and molecule biology cross comprehensive evaluation, and important guiding significances are provided for optimal research of the surface of the bone substitute implant.

The invention discloses a small-molecule inhibitor for a bone formation negative regulatory factor Smurf1. The compound shown as a formula I can be used in preparation of medicines for promoting bone formation and preparation of a BMP sensitizing agent. The invention also provides one of the applications of the compound shown as the formula I: (1) preparation of products for improving osteogenic potential of cells, wherein the cells are specifically myogenic precursor cells or osteogenic precursor cells; (2) preparation of products for improving alkaline phosphatase, osteocalcin and / or type I collagen mRNA levels in cells, wherein the cells are specifically myogenic precursor cells or osteogenic precursor cells; (3) preparation of products for improving activity of alkaline phosphatase in myogenic precursor cells or osteogenic precursor cells or osteoblasts; (4) preparation of products for inhibiting degradation of an Smad1 / 5 protein under the condition of BMP pathway activation; and (5) preparation of products for reducing ubiquitination level of the Smad1 / 5 protein under the condition of BMP pathway activation.

The invention relates to a preparation method and the application of polypeptide of fermented cucumber seeds. The method includes: firstly, preparing degreased and dried fermented cucumber seed powder; and secondly, adding the degreased and dried cucumber seed powder into buffer solution, stirring or soaking, centrifuging, performing ultrafiltration, concentrating filtrate and drying so that the polypeptide is prepared. The polypeptide of the fermented cucumber seeds is used for preventing and treating osteoporosis. The polypeptide of the fermented cucumber seeds is safe, reliable and non-toxic, is capable of promoting sclerotomal cell proliferation, lowering activity of alkaline phosphatase and improving osteoporosis symptoms, and can play a role in prevention and treatment of osteoporosis and promotion of skeleton growth.

The invention relates to a method for expressing a recombinant human bone morphogenetic protein in insect cells. The method comprises the following steps of cloning a mature peptide gene segment of the human bone morphogenetic protein (BMP) to an an improved insect cell-baculovirus protein expression vector; co-transfecting sf9 insect cells; preparing recombinant baculovirus having mature peptide gene segment of the BMP; then transfecting High5 insect cells; highly expressing the BMP mature peptides; and purifying the expressed BMP mature peptides via an Ni affinity column. Detections on in-vitro alkaline phosphatase activity and induced-bone experiments of muscular pouch embedded in mice demonstrate that both expressed BMP2 and BMP7 have activities for inducing ectopic bone formation. By using an improved insect cell-baculovirus protein expression system,, 40 mg purified BMP2 or BMP7 mature peptide can be obtained per litre cultured insect cells; and cost is reduced at least 10 times than that in CHO cells.

The invention relates to a dental implant and a preparation method thereof. The dental implant comprises a titanium substrate and a honeycomb nano titanium oxide film layer, wherein the honeycomb nano titanium oxide film layer is formed on the surface of the titanium substrate in situ to form a honeycomb nano shape on the surface of the dental implant; the oxygen content of the honeycomb nano titanium oxide film layer is reduced with increase in thickness of the film layer and is gradually transited to the titanium substrate. The honeycomb nano titanium oxide film layer is formed on the surface of the titanium substrate in situ so as to form the honeycomb nano shape on the surface of the dental implant, wherein the honeycomb nano titanium oxide film layer is thin, has better strength, is not obviously layered with the titanium substrate and is firm in combination, and the regular nanostructure can effectively reinforce the adhesion of mesenchymal stem cells to the surface of the implant and the expression of alkaline phosphatase activities and osteogenesis-related genes, and shows superior osteogenic differentiation induction capacity, so that the long-term stability and the success rate of the dental implant are improved.

The invention discloses modestobacter new species and an application thereof, and provides Modestobacter deseti CPCC D00004 which is preserved in China General Microbiological Culture Collection Center, CGMCC with the preservation No. of CGMCC No.18004. The bacterial strain CPCC D00004 has alkaline phosphatase activity and acidic phosphatase activity, can degrade organophosphor, is functional modestobacter new species, and has broad application prospects in the respects of garbage disposal, soil remediation, control of desert, soil improvement and the like.