37 results about "Mouse Nerve" patented technology

The invention discloses a method for preparing polyclonal antibody against mouse nerve growth factor (NGF) and application thereof in an enzyme-linked immunoassay method. The polyclonal antibody which is prepared by using mouse submandibular gland NGF as an immunogen can be used for quantitatively detecting the mouse growth factor. Therefore, an assembled kit can determine fixed position and expression of different tissue cell nerve growth factors and detect NGF contents in different samples such as blood, cells, tissues, culture medium supernatant, and the like.

The invention discloses a method for isolating recombinant human nerve growth factor from mammalian cell culture, comprising the steps of carrying out strong cation exchange chromatography, to be specific, sampling supernate of clarified mammalian cell culture to strong cation exchange chromatography resin, eluting with elution buffer to obtain first recombinant nerve growth factor; carrying out hydrophobic chromatography, to be specific, treating the first recombinant nerve growth factor, sampling to hydrophobic chromatography resin, and eluting with elution buffer to obtain second recombinant nerve growth factor. The method has few operation steps, the production cycle is shortened, recovery rate is increased, the purity is high, the uniformity is good, and in-vitro bioactivity is very high, generally higher than 1.5*105 U / mg, which is far higher than that (0.5*105 U / mg) of mouse nerve growth factors for injection in the market.

The invention relates to a medicament for nerve regeneration promotion, and particularly to a nerve regeneration promotion injection and a preparation method thereof. The nerve regeneration promotion injection is composed of the following ingredients in proportion: 280-320 micrograms of mouse nerve growth factor for injection, 19-21ml of single sialic acid four hexose ganglioside sodium injecta, 18-22ml of cerebroside-kinin injecta, 9-11ml of mecobalamine injecta and 9-11ml of dexamethasone acetate injecta. The five effective ingredients of the medicament provided by the invention are combined together and cooperative actions of multiple nutritive factors are utilized, so that the medicament has a significant curative effect on repairing of injured nerve, especially the repairing of spinal nerve injury. The medicament provided by the invention also has a protection effect on spinal cord transected injury at the early stage.

The invention discloses a method for preparing a mouse nerve growth factor (NGF) and a method for preparing a mouse nerve growth factor for injection. The method for preparing the mouse nerve growth factor adopts 100KD and 5KD of ultrafiltration membrane filtration and pasteurization processes on the basis of the conventional process, can effectively filter a macromolecular virus, can further inactivate potential viruses, such as pseudo rabies viruses (PRV), Sindbis viruses, encephalomyo-carditis (EMC) viruses and the like by pasteurization at the same time, does not introduce any pollutant, such as a foreign organic solvent, a detergent and the like, and can keep high purity, high specific activity and high safety of the mouse nerve growth factor.

The invention discloses a method for separating a natural sequence nerve growth factor from a mixture. A solution containing a human or mouse nerve growth factor is separated and purified through metal ion chelating chromatography to obtain the natural sequence nerve growth factor; the metal ion chelating chromatography resin comprises a supporting medium, a chelating ligand and metal ions; the metal ions are chelated on the chelating ligand; the elution condition is to change the concentration gradient of protein and metal ion coordinate bound competitive materials. Strong cation exchange chromatography can be conducted before metal ion chelating chromatography; after metal ion chelating chromatography, efficient strong cation exchange chromatography can be conducted. According to the method, the nerve growth factor with natural sequence and without any protein tags can be purified and separated rapidly.

The invention discloses a double-antibody sandwich enzyme immune gauging method of mouse nerve growth factor, which detects whether the NGF exists and the content qualitatively or quantatively.

The invention relates to the technical field of biomedical engineering, in particular to a mouse nerve growth factor (mNGF) and a crystal structure and a cell biological effect of a lipid small molecule lysoPS composite. Key amino acids which are combined by the mNGF and the lysoPS are analyzed and determined by means of the crystal structure, namely, the 88th lysine, and the locus of the 88th lysine is mutated by means of a technology of site-directed mutagenesis. The research result shows that the combination capacity of the mNGF mutant with the lysoPS declines obviously, and the toxic and side effect on cells of the mNGF is reduced significantly. The invention provides the crystal structure of the composite, the structural characteristics of the lipid small molecule lysoPS, the preparation method of the mNGF mutant proteins and functions of the mNGF mutant proteins on regulation of drugs of an immunization system and a nervous system.

The invention discloses a preparation method of a mouse meningeal single-cell suspension. The preparation method comprises the following steps of: 1) preparing a reagent and a culture medium; 2) carrying out tissue pretreatment; 3) carrying out tissue digestion and dissociation; 4) terminating digestion; 5) filtering by a micron cell filter; 6) obtaining a target tissue single-cell suspension; and 7) measuring the yield of living cells. The preparation method of the mouse meningeal single-cell suspension can be used for quickly and efficiently obtaining the mouse meningeal single-cell suspension at low cost and ensuring that dissociated cells keep good activity, can be applied to dissociation, separation and purification of mouse meningeal cells in a large range and establishment of a cell line, and provides important reference data for research of mouse nerve tissue single cells, gene functions and genetic breeding.

The present invention relates to an expression vector for expressing a growth factor and a cell, specifically, relates to a mouse nerve growth factor-containing recombinant vector and a recombinant cell, and also relates to a preparation method of the recombinant cell, a purification method of a mouse nerve growth factor and an application of the recombinant vector or the recombinant cell in preparation of the mouse nerve growth factor. The mouse nerve growth factor is successfully expressed by utilizing a mammalian cell expression system, and the mouse nerve growth factor is further proved to have correct sequence and higher specific activity, and has a good application prospect.

The invention which relates to the biomedicine field discloses a mechanical separation method of neurons of adult mice. The method comprises the following steps: making the adult mice's brain tissue slices with the thickness of 350-375mum in an ice bath tissue culture solution with an electronic vibration slicer, preserving for 30-60min at room temperature, putting the brain tissue slices into a papain-containing tissue culture solution under aseptic conditions, controlling the temperature at 32.2-35.3DEG C, maintaining the temperature for 30min, placing the brain slices in a cell culture dish containing an HEPES buffer solution, selecting a neural region needing separation, and separating neurons through using a cell separator; and screening healthy neurons. The method of the invention, which allows a primary culture chemical separation method to be integrated into mechanical cell separation and the brain slices to be preprocessed with the papain, makes the healthy neurons of adult and aged mice to be obtained through the mechanical separation, and has the advantages of simplicity, convenience, and substantial separation effect.

The invention belongs to the field of medication and particularly relates to an eye drop for treatment of nearsightedness and a preparation method of the eye drop. The eye drop comprises, by weight, 5-15 parts of silkworm larva, 5-15 parts of caulis dendrobii powder, 70-79 parts of fructus ligustri lucidi, 90-100 parts of semen cassiae, 50-80 parts of Vitamin B1, 50-70 parts of Vitamin C, 5-15 parts of borneol, 5-15 parts of mouse nerve growth factors, 50-80 parts of Vitamin B12, 5-15 parts of sodium chloride and 60-80 parts of piracetam. By the arrangement, problems that drug therapy is proneto side effects, defects prone to recurrence, risks and sequelae of surgical treatment and wearing optical glasses can only overcome the shortcoming of daily vision of patients with visual impairmentbut not the root cause are solved, and the eye drop adopts natural raw materials is developed into a safe, efficient and high-quality product with reasonable compatibility and synergistic effect, remarkable effect in the treatment of nearsightedness and safety and efficiency.

The inventions discloses a novel mouse nerve growth factor intravitreal injection drug delivery system and an application thereof, and relates to the field of mouse nerve growth factor-containing drug for treating oculopathy through intravitreal injection drug delivery. In allusion to the problems that non-ideal treatment effect and big side effects existing in a conventional drug delivery mode of a mouse nerve growth factor for treating oculopathy, the invention aims to provide the mouse nerve growth factor intravitreal injection drug delivery system which is characterized by comprising the mouse nerve growth factor and an ultrasonic contrast agent. The invention also provides the application of the mouse nerve growth factor intravitreal injection drug delivery system in preparing a drug for treating glaucomatous optic nerve damage, wherein the drug is used for being administrated by intravitreal injection under ultrasonic irradiation. The novel drug delivery mode provided by the invention has the advantages of good treatment effect, safety and reliability, and can be used for delaying or preventing occurrence and development of the glaucomatous optic nerve damage.

The invention discloses a method for extracting a nerve growth factor, wherein the method includes the steps: acidizing and centrifuging a supernatant containing the nerve growth factor to obtain an acidolysis supernatant, carrying out cation-exchange chromatography on the acidolysis supernatant, and collecting a target elution peak containing the nerve growth factor, to obtain a protein liquid 1;and carrying out hydrophobic chromatography on the protein liquid 1, and collecting a target elution peak containing the nerve growth factor, to obtain a target protein liquid. The method does not include the step of cationic reverse adsorption chromatography. Compared with the prior art, the method has simple steps and is safer and more reliable, and the purity, yield and sample activity of theobtained nerve growth factor sample are improved. The method not only is suitable for extracting the mouse nerve growth factor, but also is suitable for extracting the human nerve growth factor, and provides a new way for large-scale separation and purification of the nerve growth factor.

The invention discloses a clustering enzyme with atomic precision, preparation and application of the clustering enzyme in nervous system diseases. The clustering enzyme is prepared by the following method: preparing HAuCl4 water as a solution I; 3-mercaptopropionic acid aqueous solution as a solution II; adding the solution I into the solution II, and stirring to obtain a colorless and transparent mixed solution; continuously adding a NaOH aqueous solution and a NaBH4 solution into the reaction system; and sealing and reacting in a dark place at room temperature to obtain a brown solution, stopping the reaction, and aging the reaction solution to obtain the final clustering enzyme. The research finds that the clustering enzyme with atomic precision has ultrahigh antioxidant activity and enzyme-like selectivity, is used for early intervention of mouse nervous system diseases, and can remarkably promote wound healing and weight recovery and effectively improve later behavioral characteristics. Important reference and basis are provided for research and development of medicines for treating nervous system diseases.

The invention relates to the technical field of Tibetan medicine, in particular to application of a Tibetan medicine composition in preparation of a medicine for preventing and / or treating diabetic peripheral neuropathy, the Tibetan medicine composition comprises the following raw material medicines: turmeric, nutmeg, rhizoma nardostachyos, actinolite, liquorice, artificial musk, rhizoma zingiberis, caraway, rhizoma acori calami, pepper and soda. It is found for the first time that the Tibetan medicine composition can improve the sensitivity of nerve endings of diabetic peripheral nerve injury model mice, relieve hypoalgesia and limb numbness symptoms caused by diabetic peripheral nerve injury and improve the motor nerve conduction speed of the diabetic peripheral nerve injury model mice; therefore, the Tibetan medicine composition can prevent and / or treat diabetic peripheral nerve injury and prevent and / or treat diabetic peripheral neuropathy, and is a potential medicine for diabetic peripheral neuropathy.

The invention discloses a method for separating a natural sequence nerve growth factor from a mixture. A solution containing a human or mouse nerve growth factor is separated and purified through metal ion chelating chromatography to obtain the natural sequence nerve growth factor; the metal ion chelating chromatography resin comprises a supporting medium, a chelating ligand and metal ions; the metal ions are chelated on the chelating ligand; the elution condition is to change the concentration gradient of protein and metal ion coordinate bound competitive materials. Strong cation exchange chromatography can be conducted before metal ion chelating chromatography; after metal ion chelating chromatography, efficient strong cation exchange chromatography can be conducted. According to the method, the nerve growth factor with natural sequence and without any protein tags can be purified and separated rapidly.

Disclosed is the use of genetic means or a related inhibitor to inhibit and down-regulate the amount of PI4KIIIα protein, RBO / EFR3 / EFR3A / EFR3B membrane proteins, TTC7 protein and membrane protein complexes formed by said proteins, or related enzyme activity to promote Aβ secretion by neuronal cells, reduce Aβ accumulation within neurons, and thereby reduce AD model fruit fly and mouse nerve dysfunction.

The invention belongs to the technical field of myopia regulatory diseases, and particularly discloses a biological agent for treating myopia. The biological agent comprises a basic fibroblast growthfactor, 10-30 [mu]g of a transforming growth factor and 5-15 [mu]g of a mouse nerve growth factor, wherein the mass sum of the transforming growth factor and the mouse nerve growth factor is less thanor equal to the mass of the basic fibroblast growth factor, and the mass of the basic fibroblast growth factor is less than or equal to two times the mass sum of the transforming growth factor and the mouse nerve growth factor. The invention also discloses a preparation method and use method of the biological agent for treating the myopia. The biological agent not only can promote self-healing ofthe myopia, but also is high in safety.

The present invention discloses cell adhesion molecule 5 (ICAM5) monoclonal antibody, a preparation method and applications thereof, wherein a preservation number of the hybridoma cell strain is CGMCC No.5798. The present invention provides the ICAM5 monoclonal antibody, which can be provided for ICAM5 antigen-based immunofluorescence and Western blotting, and other scientific researches, wherein test results show that the high and stable expression exists on the surface of human and mouse nerve cells. In addition, the antibody has characteristics of strong specificity and high titer, wherein an ELISA detection titer can be 1:400000, such that an ICAM5 antibody use concentration in a test is low, and an antibody use amount is low; and the ICAM5 antigen is positioned on the surface of nerve cell bodies and nerve fibers, such that structures of nerve cell bodies and nerve fibers can be clearly displayed with immunofluorescence staining.

The invention discloses a method for intrathecally injecting a nerve regeneration promoting injection. The method is completed by injecting the nerve regeneration promoting medicine into the arachnoid membrane. The nerve regeneration promoting injection is prepared from the following components: 280-320mu g of mouse nerve growth factors for injection, 19-21ml of monosialotetrahexosyl ganglioside sodium injection, 8-22ml of cattle encephalon glycoside and ignotin injection, 9-11ml of mecobalamine injection and 9-11ml of dexamethasone acetate injection. The method is conductive to reducing secondary injury after spinal cord injury, and the intrathecal injection has an excellent biological safety.

The invention relates to a new application of caesalpinia minax, namely a new application of caesalpinia minax in preparation of medicines for treating an alzheimer's disease, in particular to a new application of a caesalpinia minax extract in preparation of medicines for treating the alzheimer's disease. The learning ability and the memory of dementia mice treated by the caesalpinia minax extract are obviously improved; the clearance rate of oxygen radicals in the brain tissue is improved; and beta-starch amyloid protein is reduced. The caesalpinia minax has a protecting effect on nerve injuries of AD mice and has positive reference for development and research and clinical treatment of medicines for clinical treatment of elder diseases of the alzheimer's disease and the like.

A mouse nerve retrograde tracing mark component comprises a cylinder tank body and an upper cover. A shape of the cylinder tank body is a cylindrical shape, a cross section of the cylinder tank body is a major arc, an upper cover is a curved plate and the cross section is an inferior arc, the upper cover fastens the cylinder tank body, and the upper cover and the cylinder tank body form a completecylindrical shape. Single and multiple retrograde tracing marking of mouse nerves can be realized so that direct observation of a mutual relation between different sources of motor neuron pools or neuronal subgroups in a spinal cord becomes possible.

The invention discloses a method for preparing polyclonal antibody against mouse nerve growth factor (NGF) and application thereof in an enzyme-linked immunoassay method. The polyclonal antibody which is prepared by using mouse submandibular gland NGF as an immunogen can be used for quantitatively detecting the mouse growth factor. Therefore, an assembled kit can determine fixed position and expression of different tissue cell nerve growth factors and detect NGF contents in different samples such as blood, cells, tissues, culture medium supernatant, and the like.

The invention relates to the technical field of medicines, in particular to application of ganoderma lucidum polysaccharide in preparation of a medicine for treating central nervous system injury, and discloses a preparation method of the ganoderma lucidum polysaccharide. The specific preparation method comprises the following steps: taking ganoderma lucidum spores as a raw material, and performing degreasing, water extraction, deproteinization, alcohol precipitation and drying to obtain ganoderma lucidum polysaccharide. Cell proliferation experiment detection shows that the ganoderma lucidum polysaccharide can promote neuronal cell proliferation and relieve neuronal damage caused by methamphetamine, and can improve mouse neurobehavioral disorder and learning and memory disorder caused by methamphetamine, reduce neuron loss and increase expression of nerve growth factors (BDNF). Therefore, the ganoderma lucidum polysaccharide has a good protection effect on neuron injury and can be used for treating central nervous system injury. According to the application of the ganoderma lucidum polysaccharide in preparation of the medicine for treating the central nervous system injury, the application field of the ganoderma lucidum polysaccharide is widened, and meanwhile, a new treatment scheme is provided for treatment of the central nervous system injury.

The present invention relates to an expression vector for expressing a growth factor and a cell, specifically, relates to a mouse nerve growth factor-containing recombinant vector and a recombinant cell, and also relates to a preparation method of the recombinant cell, a purification method of a mouse nerve growth factor and an application of the recombinant vector or the recombinant cell in preparation of the mouse nerve growth factor. The mouse nerve growth factor is successfully expressed by utilizing a mammalian cell expression system, and the mouse nerve growth factor is further proved to have correct sequence and higher specific activity, and has a good application prospect.

The invention discloses a monoclonal antibody against a human neurofilament medium molecular weight subunit and a preparation method and application thereof. The monoclonal antibody has specificity for polypeptide with an amino acid sequence shown in SEQ ID NO.1 and is produced by secretion of a cell strain with the preservation number CGMCC No. 6064. The invention also provides a hybridoma cell strain for production of the monoclonal antibody, which is named as 2C1 and has been preserved in China General Microbiological Culture Collection Center on April 27, 2012, and the preservation number is CGMCC No. 6064. The monoclonal antibody has high sensitivity, high specificity and cross-reactivity with neurofilaments of mouse and rats, and can be applied in research of detection kits for neurofilament medium molecular weight subunits of human beings, mouse and rats.