37 results about "Mouse tissue" patented technology

This application describes the cloning of p63, a gene at chromosome 3q27-29, that bears homology to the tumor suppressor p53. The p63 gene encodes at least six different isotypes. p63 was detected in a variety of human and mouse tissue and demonstrates remarkably divergent activities, such as the ability to transactivate p53 reporter genes and induce apoptosis. Isotopes of p63 lacking a transactivation domain act as dominant negatives towards the transactivation by p53 and p63.

The invention relates to an immune magnetosome for detecting Bt insecticidal protein in mice tissue and a preparation method thereof. The immune magnetosome is coated by anti-Bt insecticidal protein polyclonal antibody with the particle size being 50nm. The preparation method of the immune magnetosome comprises the following steps of: performing ultrasonic disruption and magnetic adsorption to obtain purified magnetosome of magnetotactic bacteria, mixing adjuvant and Bt insecticidal protein, immunizing mice, performing separation and purification to obtain mouse anti Bt insecticidal protein polyclonal antibody and coupling the antibody and the magnetosome by a carboxy-amino chemical coupling method to obtain the immune magnetosome for detecting Bt insecticidal protein in mice tissue. By chemiluminescence EIA detection, the immune magnetosome provided by the invention has a luminous unit of 83kcounts / microgram magnetosome. The correlation of luminous intensity and Bt protein concentration shows that as the protein concentration goes up, the luminous intensity goes down with a range being 1-103ng / ml and with a minimum detectable concentration being 1ng / ml. The method for the practical detection of Bt insecticidal protein content in mice tissue has advantages of high sensitivity, stable result and simple operation, and shows a good application prospect.

The invention discloses a brain targeted liposome preparation of a <99m>Tc tumor imaging medicament and application thereof. In the invention, the idea that an active brain targeted liposome entraps the <99m>Tc tumor imaging medicament is put forward, and the brain targeted liposome preparation of the <99m>Tc tumor imaging medicament is prepared by a fast film dispersion method, wherein liposome has the entrapment rate of over 99 percent, the particle size of less than 30 nm and a structure shown in figure 1. Phospholipid is dispersed in water to form single-layer or multi-layer microcapsules; the liposoluble <99m>Tc medicament is inserted into a bimolecular lipid membrane; and a bradykinin analogue RMP-7 is spread on the surface of the liposome as an active brain targeted directing molecule. Mouse tissue distribution experiments show that the preparation greatly improves the capability of the <99m>Tc medicament in passing through BBB; the SPECT imaging and self-development results of a model of mice with C6 brain gliomas show that the active targeted liposome preparation can realize the early diagnosis of brain tumors when applied to the <99m>Tc tumor imaging medicament; and the brain targeted liposome preparation belongs to the technical field of radiopharmacy and medicinal preparations.

Monoclonal antibodies against human and mouse tissue factor, and fragments thereof, are disclosed, as well as pharmaceutical compositions and compositions for drug delivery containing the same. Therapeutic methods using the same are also disclosed.

The invention relates to a method for constructing a mouse TCR alpha CDR3 region sequencing library. The method mainly comprises the following steps: (1) extracting total RNA from mouse tissues or whole blood; (2) reversely transcribing the RNA into cDNA, ligating a linker sequence to the end of the long-stranded cDNA, and amplifying TCR alpha containing a CDR3 region through a universal C-terminal primer and a linker sequence primer; (3) fragmenting the TCR alpha through Tn5 transposase, and enriching a CDR3 region sequence; and (4) carrying out sequencing by an illumina high-throughput sequencing platform. The method is suitable for library construction of tissues containing various types of T cells, and body fluids, can highly-efficiently amplify the mouse TCR alpha through a 5'RACE technology in an agonic manner, and solves the problems of difficulty in control of the template copy number and the offset of the product, caused by multiplex PCR amplification; and the Tn5 enzyme's ability to simultaneously complete DNA fragmentation and linker ligation and a specific primer are used to fast and accurately enrich and construct a library of the hypervariable region (CDR3 region) ofthe TCR alpha, so pertinent sequencing, easy data analysis and sequencing cost saving are achieved.

The invention discloses a method for detecting compound genotoxicity by a biomarker and a use of the biomarker. The biomarker is a mice BC gene. The method comprises the following steps of 1, treating a mice or cells by a compound to obtain mice tissue or a cell sample, and 2, detecting the expression of a biomarker in the mice tissue or the cell sample, wherein the biomarker is a mice BC gene and through the expression of the biomarker, genotoxicity of the compound is detected. The method for detecting compound genotoxicity has the characteristics of accuracy, high sensitivity and easy operation, can be used as a candidate method for evaluating drug safety and can be used for early screening of genotoxicity of a new drug.

The invention provides a preparation method and application of an anti-oxidation active component of bamboo willows. The preparation method takes bamboo willow peels as raw materials and comprises the following steps: carrying out ultrasonic extraction for 1-5 times by utilizing 5 times to 20 times of ethanol with the concentration of 70% to obtain an extracting solution, wherein each time of extraction is carried out for 30min-120min; decompressing and concentrating the extracting solution until no ethanol is remained; adding distilled water and dissolving; extracting by utilizing ethyl acetate and water saturated n-butanol; decompressing, concentrating and drying an n-butanol extracting solution to obtain crude extract powder; and purifying the crude extract powder to obtain the anti-oxidation active component of the bamboo willows. The active component has a very strong cleaning effect on ABTS (2,2'-Azinobis-(3-ethylbenzthiazoline-6-sulphonate))+.free radicals and DPPH (1,1-diphenyl-2-picrylhydrazyl)+.free radicals; the activity of superoxide dismutase (SOD) in mouse tissues can be improved; and the damages to free radicals are inhibited and lipid peroxidation is prevented. A human body test shows that the active component can be directly used as a prescribed preparation for treating diseases, such as hyperlipidemia, caused by the fact that the free radicals in the human body are excessive. According to the preparation method and application of the anti-oxidation active component of bamboo willows, the anti-oxidation active component is successfully extracted from the bamboo willows and a foundation is laid for application of the bamboo willows to the field of medical products.

Disclosed is an evaluation method which can rapidly discriminate a Dao− / − homozygote from a large number of animals produced in a mating experiment between a DAO enzyme deficient mouse and other disease model mice, to rapidly perform a quantitative measurement of the D-amino acids contained in a large number of samples. The invention provides a method for evaluating the effect of a test condition on a mouse tissue, or cultured tissue cells derived from the tissue. The method comprises the steps of: providing a Dao1− / − mouse or the like; exposing the tissue from the Dao1− / − mouse or the like, to the test condition; and analyzing the effect of exposing the tissue from the Dao1− / − mouse or the like, to the test condition.

The invention discloses a method for constructing a mouse model capable of specifically knocking out an IKKalpha gene in a hippocampus region, a targeting vector and a kit. The IKKalpha gene is knocked out from specific mouse tissues through utilizing a Cre-LoxP recombination system, so that a transgenic mouse with the knocked-out IKKalpha gene is established, and an NF-omicronB signal channel and an acting mechanism on diseases, particularly tumors, are researched on the whole level of animals. Meanwhile, in order to determine and monitor whether specific knockout of the IKKalpha gene in the hippocampus region causes influences on the behavioral science of the mouse, red luciferase RFP720 is connected into a transgenic carrier. A gene targeting technology is adopted, and expression of the endogenous IKKalpha gene in the hippocampus region of the mouse is reduced based on iRFP720 near-infrared fluorescent protein and a Cre-LoxP gene knockout system, so that living body imaging of the transgenic mouse is monitored through an animal imaging system.

The invention relates to a quick identification method of genetically modified mouse genotypes. The quick identification method comprises two kinds of autonomous research and development lysate SG1 Buffer and SG2 Buffer, and comprises the steps of obtaining the DNA of genetically modified mouse tissue, amplifying target DNA fragments and performing gel electrophoresis on PCR products. The autonomous research and development tissue lysate can realize digestion of mouse tail tissue in water bath or metal bath of 95 DEG C for 5-10min, the genome DNA in the mouse tissue is rapidly released, wherein pyrolysis products are free from extraction and purification, and are directly used as a PCR amplification formwork, so that the total time for preparing the genetically modified mouse identification formwork is shortened to 10-15min. A PCR premixing solution 2*PCR Master Mix ( With Dye ) is used, experiment systems and conditions are optimized by the method, the target DNA fragments are amplified, then the PCR products are subjected to gel electrophoresis, and the identification results are quickly obtained.

The invention discloses a method for extracting mycobacterium tuberculosis DNA from a mouse tissue. The method for extracting the mycobacterium tuberculosis DNA from the mouse tissue comprises the following steps: (1) homogenising a mouse lung tissue, centrifuging, then abandoning a supernatant, adding a GA buffer solution and a proteinase K solution, and then carrying out water bath for implementing histolysis, so as to obtain cell suspension; (2) adding a TES buffer solution (containing lysozyme) and an SDS solution into the cell suspension, putting the cell suspension in a boiling water bath for 60 minutes, and placing on ice for obtaining suspension; and (3) adding a sodium acetate solution and a glucose solution into the suspension, placing the suspension on ice, then adding a phenol / chloroform / isoamyl alcohol solution for extracting, centrifuging and then taking an upper water phase, adding chloroform / isoamyl alcohol into the upper water phase for extracting, centrifuging, then taking an upper water phase, adding ethyl alcohol, oscillating and then centrifuging, abandoning a supernatant, and airing, thereby obtaining the total DNA. The method for extracting the mycobacterium tuberculosis DNA from the mouse tissue is high in extraction ratio and easy to operate, and the purity and concentration of the obtained DNA are high.

The invention discloses a preparation method of mouse ECMs (Extracellular Matrixes), obtained mouse ECMs and a mouse ovarium in-vivo regeneration method. The method comprises the following steps of 1, performing treatment for different time by using detergents with different concentrations, removing cells of mouse tissue organs, and preparing the ECMs; 2, performing histological analysis and biochemical analysis on the prepared ECMs so as to determine the optimum treatment condition; and 3, performing orthotopic transplantation on the ECMs subjected to cell removal treatment, recording the development condition of the ECMs and the ovarium regeneration efficiency, and further, verifying the regenerated ovarium tissues by a histology method, an immunohistochemistry method, a PCR (Polymerase Chain Reaction) method and the like. In addition, the effect that the tissue scaffolds (ECM) from different sources can also promote the ovarium regeneration is also proved. By using the method provided by the invention, the ovarium is successfully regenerated for the first time; and the further solving method is provided for the problem of female apogeny due to ovarium dysfunction. The method provided by the invention has the advantages of simplicity, convenience, low cost, low experiment condition requirements and high operability.

Monoclonal antibodies against human and mouse tissue factor, and fragments thereof, are disclosed, as well as pharmaceutical compositions and compositions for drug delivery containing the same. Therapeutic methods using the same are also disclosed.

The invention discloses a preparation method of a cold shock protein. The preparation method is characterized by comprising the following steps of 1, TRIzol method-based total RNAs extraction: grinding a mouse tissue sample by a mortar and adding 1ml of a pyrolysis liquid into each 50-100mg of the tissue, 2, cDNA first-chain synthesis: adding 5 microliters of a RNA solution, 1 microliter of Oligo and 1 microliter of dNTP into 20 microliters of a reaction system and carrying out a reaction process at a temperature of 65 DEG C for 5min, 3, a reaction process: adding 1 microliter of cDNA, 2 microliters of dNTP, 1 microliter of TapDNA polymerase, 1 microliter of an upstream primer, 1 microliter of an downstream primer and 2 microliters of 10*buffer into 20 microliters of the reaction system, 5, inducible expression, 6, preliminary purification and 7, high-degree purification. The cold shock protein which is coldinducible RNA-binding protein (CIRP) has effects of protecting nervous tissue cells and resisting nervous tissue cell apoptosis.

The invention discloses a preparation method of a cold shock protein. The preparation method is characterized by comprising the following steps of 1, TRIzol method-based total RNAs extraction: grinding a mouse tissue sample by a mortar and adding 1ml of a pyrolysis liquid into each 50-100mg of the tissue, 2, cDNA first-chain synthesis: adding 5 microliters of a RNA solution, 1 microliter of Oligo and 1 microliter of dNTP into 20 microliters of a reaction system and carrying out a reaction process at a temperature of 65 DEG C for 5min, 3, a reaction process: adding 1 microliter of cDNA, 2 microliters of dNTP, 1 microliter of TapDNA polymerase, 1 microliter of an upstream primer, 1 microliter of an downstream primer and 2 microliters of 10*buffer into 20 microliters of the reaction system, 5, inducible expression, 6, preliminary purification and 7, high-degree purification. The cold shock protein which is coldinducible RNA-binding protein (CIRP) has effects of protecting nervous tissue cells and resisting nervous tissue cell apoptosis.

The invention relates to a novel coronavirus COVID-19 fluorescent quantitative PCR (Polymerase Chain Reaction) detection primer, a probe, a kit and a detection method, and the novel coronavirus COVID-19 fluorescent quantitative PCR detection primer and probe comprise specific primers aiming at a novel coronavirus COVID-19 N gene as shown in SEQ ID NO.1-2, and a first probe as shown in SEQ ID NO.3. The novel coronavirus COVID-19 detection kit provided by the invention can rapidly, accurately and sensitively detect the novel coronavirus COVID-19 in mouse tissues, the detection sensitivity is as high as 10 copies / mL, quantitative analysis can be carried out while qualitative analysis is carried out on viruses in the mouse tissues, and the quantitative linear range is good. The movel coronavirus COVID-19 fluorescent quantitative PCR detection primer and probe, the kit and the detection methodare used for detecting the novel coronavirus COVID-19 in laboratory mouse tissues, and have a relatively high application value.

The invention provides an anti-SETD3 monoclonal antibody and use thereof. The monoclonal antibody can recognize an SETD3 protein and comprises a heavy chain variable region and a light chain variable region, the heavy chain variable region has three complementary determining regions of amino acid sequences as shown in SEQ ID NO: 1-SEQ ID NO: 3; and the light chain variable region has three complementary determining regions of amino acid sequences as shown in SEQ ID NO: 4-SEQ ID NO: 6. The anti-SETD3 monoclonal antibody has good recognition specificity on an in-vitro purified protein, in-vitro cultured mammalian cell endogenous and exogenous SETD3 proteins and a mouse tissue SETD3 protein. Compared with a commercially available antibody, the monoclonal antibody has higher recognition specificity.

The invention discloses a novel application of 7-methoxyl baicalein, namely an application in preparing anti-hypoxic medicines. The integral animal hypoxic experiments show that 7-methoxyl baicalein has a remarkable anti-hypoxic activity, and not only can prolong the survival time of a closed hypoxic mouse at a normal pressure, but also can prolong the survival time of an acute decompression hypoxic mouse. The mechanism of action of 7-methoxyl baicalein may be related to effective removal of excessive free radicals generated in hypoxic mouse tissues and improvement of oxidation resistance.

The invention discloses an application of wedelolactone in preparation of a medicine and a health care product for preventing and / or treating herpes virus infection. Experiments prove that wedelolactone has an inhibitory activity effect on herpes simplex virus infection, that is, wedelolactone has anti-HSV-1 and HSV-2 activity, and is relatively low in toxicity. Wedelolactone can significantly inhibit expression of related proteins and mRNA of HSV-1 and HSV-2, has obvious dose dependence, and has the best effect at the early stage after HSV infection, namely 2-4 h. Wedelolactone achieves the effect of blocking herpes virus infection by directly inactivating virus particles or intervening the replication process after viruses enter cells and inhibiting virus proliferation, and the wedelolactone can reduce the virus load in mouse tissues on the in-vivo animal level. Therefore, wedelolactone has the potential of being developed into a novel anti-HSV preparation of targeted virus particles.

The invention discloses a method for detecting the number of human cells implanted in a mouse and a kit utilizing the method. The kit utilizing the method for detecting the number of human cells implanted in a mouse comprises a primer pair for amplifying a human Down's syndrome DNA sequence fragment. The human Down's syndrome DNA sequence fragment has the sequence 3 in the sequence table. The method and the kit realize specific detection on the number of human cells in mouse tissue. Through use of the primers having high specificity, the kit has the advantages of high specificity, strong sensibility and good repeatability.

The invention discloses a method for detecting the number of human cells implanted in a mouse and a kit utilizing the method. The kit utilizing the method for detecting the number of human cells implanted in a mouse comprises a primer pair for amplifying a human Down's syndrome DNA sequence fragment. The human Down's syndrome DNA sequence fragment has the sequence 3 in the sequence table. The method and the kit realize specific detection on the number of human cells in mouse tissue. Through use of the primers having high specificity, the kit has the advantages of high specificity, strong sensibility and good repeatability.

The invention provides a method for preparing and purifying recombinant adenovirus expressing Sox4 gene, and belongs to the field of genetic engineering. The method connects an Sox4 gene fragment withan adenovirus pAdtrack-CMV carrier to obtain a recombinant adenovirus expression plasmid pAdtrack-Sox4. Recon formed by linearized pAdtrack-Sox4 and an AdEasy carrier transfects HEK293 cells to generate adenovirus. The virus is purified by gradient centrifugation of cesium chloride, the virus titer is detected by a micro total cell pathological change method, and finally, the whole process of adenovirus packaging, production and purification is realized. The recombinant adenovirus of Sox4 gene can successfully infect 293T cells and mouse inguinal adipose tissues to realize continuous high expression of Sox4 gene in somatic cells and mouse tissues, which lays a good technical foundation for future research on the function of the Sox4 gene.

The invention discloses a protein plasmid for expressing mouse p53a or p53b protein as well as a construction method and application thereof, and the construction method comprises the following steps: extracting RNA (Ribonucleic Acid) in mouse tissues, and carrying out reverse transcription to obtain cDNA (Complementary Deoxyribonucleic Acid); respectively carrying out PCR (Polymerase Chain Reaction) amplification on genes for expressing p53a and p53b proteins by taking the cDNA as a template; carrying out BamH I and EcoR I double enzyme digestion on a gene segment expressing p53a protein, a gene segment expressing p53b protein and pCMV-HA, recovering enzyme digestion products, connecting and transforming escherichia coli, and extracting recombinant plasmids; carrying out transfection amplification on recombinant plasmids which are subjected to enzyme digestion identification and correct sequencing, and extracting plasmids; the construction method has the characteristics of rapidness, accuracy and high efficiency.

Disclosed is an evaluation method which can rapidly discriminate a Dao− / − homozygote from a large number of animals produced in a mating experiment between a DAO enzyme deficient mouse and other disease model mice, to rapidly perform a quantitative measurement of the D-amino acids contained in a large number of samples. The invention provides a method for evaluating the effect of a test condition on a mouse tissue, or cultured tissue cells derived from the tissue. The method comprises the steps of: providing a Dao1− / − mouse or the like; exposing the tissue from the Dao1− / − mouse or the like, to the test condition; and analyzing the effect of exposing the tissue from the Dao1− / − mouse or the like, to the test condition.

The invention discloses a DNA extraction kit applied to transgenic mouse gene identification. The DNA extraction kit comprises a tissue lysis solution, protease K, a binding solution, adsorption microspheres, a washing solution and an eluent, wherein the adsorption microspheres are silicon dioxide microspheres with the diameter of 50 to 100 [mu]m. Compared with a traditional kit, the DNA extraction kit disclosed by the invention has the advantages of low price, few operation steps and high extraction purity; used reagents are non-toxic, safe and stable; and the DNA extraction kit is suitable for extracting DNA of tail tips and toes of mice on a large scale. The invention also discloses a method for performing high-throughput extraction on mouse tissue genome DNA by using the kit. The provided extraction method has the advantages that the operation process is simplified, the experiment time is shortened, and the quality of the extracted DNA completely meets the requirements of subsequent PCR. The transgenic mouse gene identification efficiency is obviously improved.

The invention discloses a method for constructing a mouse model capable of specifically knocking out an IKKalpha gene in a hippocampus region, a targeting vector and a kit. The IKKalpha gene is knocked out from specific mouse tissues through utilizing a Cre-LoxP recombination system, so that a transgenic mouse with the knocked-out IKKalpha gene is established, and an NF-omicronB signal channel and an acting mechanism on diseases, particularly tumors, are researched on the whole level of animals. Meanwhile, in order to determine and monitor whether specific knockout of the IKKalpha gene in the hippocampus region causes influences on the behavioral science of the mouse, red luciferase RFP720 is connected into a transgenic carrier. A gene targeting technology is adopted, and expression of the endogenous IKKalpha gene in the hippocampus region of the mouse is reduced based on iRFP720 near-infrared fluorescent protein and a Cre-LoxP gene knockout system, so that living body imaging of the transgenic mouse is monitored through an animal imaging system.

The invention provides an application of piceatannol in preparation of a medicine for preventing and / or treating herpes simplex virus infection. Experiments prove that piceatannol has an inhibitory activity effect on herpes simplex virus infection, and the piceatannol has anti-HSV-1 and HSV-2 activity and is extremely low in toxicity. After piceatannol treatment, virus protein expression of HSV-1 and HSV-2 can be significantly inhibited, and obvious dose dependence is presented. The piceatannol achieves the effect of resisting herpes simplex virus infection by directly inactivating virus particles or intervening a replication process after a virus enters cells, so that virus proliferation is inhibited, the virus load in animal mouse tissues is reduced, the physical condition of a mouse is improved, and the body index of the mouse is increased, and therefore, the piceatannol has the potential of being prepared into the medicine for preventing and / or treating the herpes simplex virus.