56 results about "Na k atpase activity" patented technology

The invention relates to a dental implant and a preparation method of a bioactive antibacterial surface of the dental implant. The dental implant is characterized in that medical titanium or a titanium alloy is taken a substrate; a nanotube morphology is constructed on the surface of the substrate through anodic oxidation; the surface of the substrate is uniformly coated with a polydopamine bionic membrane layer formed by auto-polymerization; the dental implant with the bioactive antibacterial surface provided with in-situ grafted silver nanoparticles is obtained by means of the adsorption and reduction capabilities of a polydopamine surface for silver ions. By adopting the dental implant, a uniform discrete nanotube oxide layer is formed in situ on the surface of the titanium substrate to be taken as a storage carrier of a large quantity of antibacterial silver particles, and the bioactive antibacterial surface which grows in situ is prepared through the steps of surface dopamine auto-polymerization and in-situ soaking and reduction and the like, so that the implant surface has more durable antibacterial property, the hydrophilic performance and bioactivity of the surface are improved, the alkaline phosphatase activity of osteoblast on the implant surface is increased, the differentiation of the osteoblast is facilitated, the infection risk of the implant after the implant is implanted is lowered, and the implanting success rate is increased.

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 present invention provides a rat embryonic stem cell characterized by having the following properties of (a) expressing Oct3 / 4 gene and Nanog gene, (b) positive for alkaline phosphatase activity, (c) having an embryoid body forming ability, (d) expressing SSEA (Stage-Specific Embryonic Antigen)-1 and SSEA-4, (e) having the same number of chromosomes as does a normal rat cell, (f) capable of being subcultured and holding the undifferentiated state, (g) having in vitro pluripotency, (h) having a potential to differentiate for cells of three embryonic germ lineages, (i) having teratoma formation ability, and (j) having an ability to produce a chimeric rat, a method of establishing the aforementioned rat embryonic stem cell and the like.

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 invention provides a method for colorimetric-fluorescent two-channel detection of alkaline phosphatase activities. The method comprises the following steps: catalyzing a substrate trisodium L-ascorbyl-2-phosphate (AAP) of alkaline phosphatase by utilizing the alkaline phosphatase so as to hydrolyze to generate ascorbic acid (AA) with reducibility, whereas the AA can cause a Tollens reagent tobe reduced to an Ag elementary substance; under the condition that gold nanoparticles (AuNPs) and graphene quantum dots (GQDs) exist, Ag can be deposited on the surface of AuNPs to form gold and silver core shell nanoparticles (Au@AgNPs) and quenches the fluorescence of GQDs. According to the method, metallization of enzymatic silver ions, AuNPs-induced silver deposition and Au@AgNPs-quenched GQDsfluorescence are combined, the changes of color, ultraviolet-visible absorption spectroscopy and fluorescence intensity are utilized, and the colorimetric-fluorescent two-channel detection of the alkaline phosphatase activities can be realized. The method has the characterics that the sensitivity is high, the specificity is strong, the response is fast, the operation is simple, and no complex instruments are required.

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 relates to a fluorescence detection method for the activity of alkaline phosphatase, which is characterized in that sodium 4-hydroxyphenyl phosphate (4-HPP) is used as a substrate, molecules of the substrate are hydrolyzed into 1,4-benzenediol (HQ) under the catalysis of alkaline phosphatase in an aqueous solution, 1,4-benzenediol can react with subsequently added 3-(2-aminoethylamino)propyltrimethoxysilane (DAMO) to generate silicon nanoparticles (SiNPs) capable of emitting green fluorescence. The higher the activity of alkaline phosphatase in the system is, the more the generated fluorescent silicon nanoparticles are, and the higher the fluorescence intensity is. Quantitative analysis and detection of the activity of alkaline phosphatase are carried out according to the fluorescence intensity change of the detection solution. Compared with the prior art, the method has the advantages that required reagents and medicines are simple and easy to obtain, the operation is convenient, quick and accurate, and a sensitive and reliable method is provided for quantitative detection of the activity of alkaline phosphatase in a clinical sample. According to the fluorescence detection method, the detection limit is 0.1mU / mL, and the fluorescence detection method has the advantages of high sensitivity, high selectivity, wide linear range and the like.

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 relates to a brittleness grading method for crisped grass carp based on living body detection, comprising the following steps of: (1) collecting blood samples of crisped grass carp fromdifferent embrittlement stages and blood samples and muscle samples of grass carp in the same batch, pretreating the muscles, and tasting and identifying the brittleness level; (2) detecting alkalinephosphatase activity, complement C3 content, and lymphocyte content in blood samples of blood samples, and establishing a blood index database of crisped grass carp mites; (3) Establishing a non-standardized Bayes discriminant function based on blood indicators; (4) using the Bayes discriminant function to determine the brittleness level of the muscles of the crisped grass carp to be determined. The brittleness grading method for crisped grass carp based on living body detection realizes a method for detecting the brittleness of crisped grass carp by live fish, which can reflect and evaluate the embrittlement effect of crisped grass carp with objective and high accuracy without killing fish.

The invention discloses application of aloin in improvement of osteogenic capability of a bone mesenchymal stem cell (BMSC). According to the application of the invention, the BMSC is treated by aloins with different concentrations, and cell viability of the BMSC is detected; influence of the aloin on osteoblast differentiation of the BMSC is evaluated by adopting alkaline phosphatase activity andmineralization experiments; real-time fluorescent quantitative PCR detection is carried out for detecting expression situation of osteogenesis-related genes such as Runx2, ALP, OCN and OPN; variationof an ERK1 / 2-Runx2 signal channel is detected by virtue of a Western blotting process and an immumofluorescence process; and results show that the aloin promotes the osteoblast differentiation of theBMSCs by activating the ERK1 / 2-Runx2 signal channel, the osteogenic capability of the BMSCs is improved, and the research results provide data support for research and development of a new drug usedfor preventing and treating osteoporosis, so that the aloin has a greater application prospect.

The invention relates to application of miR-216 in preparation of medicines for promoting osteogenesis differentiation. The miR-216a is capable of promoting osteoblastic differentiation of human adipose tissue-derived mesenchymal stem cells (hAMSCs). Alkaline phosphatase staining, alkaline phosphatase activity, alizarin red staining and analysis of osteogenesis-related marker genes and transcription factors through RT-PCR and western blot show that the osteogenic capability of the hAMSCs is increased by the miR-216a.

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 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 present invention provides a rat embryonic stem cell characterized by having the following properties of (a) expressing Oct3 / 4 gene and Nanog gene, (b) positive for alkaline phosphatase activity, (c) having an embryoid body forming ability, (d) expressing SSEA (Stage-Specific Embryonic Antigen)-1 and SSEA-4, (e) having the same number of chromosomes as does a normal rat cell, (f) capable of being subcultured and holding the undifferentiated state, (g) having in vitro pluripotency, (h) having a potential to differentiate for cells of three embryonic germ lineages, (i) having teratoma formation ability, and (j) having an ability to produce a chimeric rat, a method of establishing the aforementioned rat embryonic stem cell and the like.

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 discloses an instant detection method for the activity of alkaline phosphatase. Through a mechanism that Cu<2+> which is subsequently added is not subjected to complexion through the pyrophosphoric acid so as to be subjected to high-sensitivity detection through a Cu<2+> selective detection electrode after an alkaline phosphatase analyte in sample solution catalyzes pyrophosphoric acid to be subjected to hydrolysis reaction, the magnitude of an electrode response signal and the activity of alkaline phosphatase in the sample solution are in positive correlation, i.e., the instantquantitative detection of the activity of the alkaline phosphatase is finished. The method disclosed by the invention is simple and easy in implementation, and an experiment can be carried out withoutoperation personnel who do not receive professional training. In addition, large professional analysis instruments, including enzyme-labeled instruments, luminoscopes, electrochemical workstations and the like, do not need to be used for carrying out quantitative signal reading, analysis cost is greatly lowered, and the method has a wide application prospect.

The invention provides application of long chain non-coded RNA-NKILA in bone tissue damage repair. Experiments verify that overexpressed NKILA can promote calcium ion deposition in a mesenchymal stemcell obviously, enhance the activity of alkaline phosphatase in the mesenchymal stem cell obviously and improve expression quantities of osteogenic differentiation markers RUNX2, SP7 and SPP1 genes inthe mesenchymal stem cell obviously.