87 results about "C57BL/6 Mouse" patented technology

The invention relates to the establishment of an animal model of non-alcoholic fatty liver disease, in particular to the establishment of a non-alcoholic fatty liver animal model induced by high fat accompanied by severe blood fat abnormality. SPF-grade male AopE- / - mice and C57BL / 6 mice were selected, randomly divided into model group and control group according to the species of mice, a total of 4 groups, 8 mice in each group, and all mice were adaptively fed with ordinary maintenance feed for 1 week. , the model group was given high-fat feed, and the control group was given control feed. After 10 weeks, the blood was drawn and sacrificed, and the four items of routine blood lipids and liver function were tested; Liver tissue was made into paraffin and frozen sections, and H.E. and oil red O staining were performed respectively to determine the effect and quality of the NAFLD model. The present invention uses a simple high-fat and high-calorie diet to make models, and creates a severe NAFLD model within a very short period of 10 weeks. The model's blood lipids, liver fat and other indicators have excellent stability and high repeatability, which is a high-quality NAFLD model. Establishment provides a reliable method.

The invention relates to a method for preparing non-alcohol fatty hepatic disease mouse model, which comprises that randomly distributing the SPF male C57BL / 6 mouse at 18-25g into model group and normal contrast group, while each group has 36 mouse; two groups can be divided into six groups via different feeding times, as 1 week, 2, weeks, 3 weeks, 4 weeks, 5 weeks, and 8 weeks, while each time point has six mouse; all mouse are fed via normal contrast forage, then to be fed via bursine-acimetion lack forage, while the contrast group is fed via normal contrast forage; weighting them in each week, killing at first, second, third, fourth, fifth, and eighth week, to evaluate the model condition. The inventive model has high stability, integral disease spectrum, and high speed.

The invention discloses a modeling method of an Sjogren syndrome mouse model. The modeling method comprises the following steps: killing mice, taking out bilateral salivary glands and peeling off capsules and connective tissues; washing with PBS (Poly Butylenes Succinate); adding the PBS according to the amount of adding 0.5ml of the PBS into each salivary gland; shearing the salivary glands into pieces, and uniformly homogenizing and centrifuging in an ice bath; then taking supernatant and quantifying salivary gland antigens by adopting a BCA (Bicinchoninic Acid) protein quantifying method; adjusting the concentration of the salivary gland antigens to be 4mg / ml by utilizing the PBS; adding equal quantity of an FCA (Freund Complete Adjuvant) or an FIA (Freund Incomplete Adjuvant) and diluting the concentration to be 2mg / ml; repeatedly blowing and beating until two liquid phases are dissolved mutually to form an ivory color; randomly grouping C57BL / 6 mice and shaving off furs on the backs of the mice; carrying out intradermal multi-site injection of 2mg / ml mouse salivary gland antigens prepared by the FCA on the back and tails of the mice on the current day and the 7th day, wherein the injection amount is 0.1ml per mouse; injecting equal quantity of the salivary gland antigens prepared by the FIA on the 14th day through the same method; after modeling for about 6 weeks, detecting indexes and screening the successfully modeled mice. The modeling method disclosed by the invention is high in modeling efficiency and short in modeling time.

The invention relates to polymethoxyflavone for treating melanoma. The effective ingredients of the polymethoxylflavone comprise one or multiple of hesperetin, nobiletin, sinensetin, 5-hydroxy-3,6,7,3',4'-pentamethoxylflavone, 5-hydroxy-3,6,7,8,3',4'-hexamethoxyflavone, and 5-hydroxy-6,7,4'-trimethoxylflavone. The polymethoxylflavone has the advantages that after the polymethoxylflavone acts for 48h, the survival rates of melanoma cell lines A2058 and A375 are respectively 42% and 26%; compared with a contrast group, the inhibition action of the polymethoxylflavone on the tumor growth in a C57b1 / 6 rat inoculated with the melanoma cell lines A375 is increased by 2.75 times.

The present invention relates to the field of biotechnology and in particular to a construction method of an experimental animal model, specifically a mouse model, with non-alcoholic steatohepatitis (NASH) transforming into hepatic carcinoma (HCC), and use thereof. The present invention provides a construction method of a mouse model with NASH and chronic steatohepatitis developing into chronic hepatic fibrosis, cirrhosis and HCC. The construction method comprises the following steps: newborn C57BL / 6 mice (including male and female newborn mice) are subjected to subcutaneous single injection of streptozotocin (STZ) (100-400 [mu]g / newborn mouse) and the mice begin to be fed with experiment use high-fat feed after the completion of female mouse breastfeeding to construct the NASH mouse model with hepatic fibrosis, cirrhosis and HCC. The provided mouse model is relatively simple in construction process, relatively simple in technical means, and low in construction cost, has an extremely high value as the needed animal model in scientific research and drug research and development, and belongs to a domestic initiative.

The invention provides a method for building a dextran sodium sulphate induced enteritis mouse model. The method includes the following steps: A, choosing female C57BL / 6 mice with the weight of 19-21g as the objects to build the mouse model; B, dissolving dextran sulphate sodium salt solution in disinfected drinking water to prepare dextran sulphate sodium salt solution with the weight percentage concentration of 2%, and placing the solution in a water bottle for protecting away from the sun, wherein the solution is prepared when required; C, feeding the chosen C57BL / 6 mice in a storageprotect feature (SPF) level animal house, enabling the mice to drink the dextran sulphate sodium salt solution prepared in the step B for six days and then drink the normal drinking water for four days. Accordingly, the chosen C57BL / 6 mice are dextran sodium sulphate induced enteritis mouse models. By means of the method for building the dextran sodium sulphate induced enteritis mouse model, death rate of animals is reduced to be zero, and pathology expressions are the same, so that the model is stable, reliable and repeatable.

A mouse animal model C57BL / 6 for the APP gene of human presenile dementia (AD) is disclosed, which can be used to research the AD, develop its medicine and screen its vaccine. Its configuring process includes using PDGF promoter to regulate APP and configure its transgene, injecting it in the male pronucleus of fertilized ovum of mouse, PCR, southern discrimination to obtain transgenic male animal, cesarean section, and DNA recombination-inbred-line passage for configuring its line.

The invention discloses a modeling method of a helicobacter pylori infection animal model. The modeling method comprises the steps of: 1, by using a 4-weeks old female C57BL / 6 mouse as the experimental animal, performing intra-gastric administration for 3 to 7 days by mixed antibiotic, wherein the mixed antibiotic is composed of gentamicin, vancomycin and nystatin; 2, raising the C57BL / 6 mouse which has been subjected to the intra-gastric administration in the step 1, for 6 to 8 days; 3, inoculating helicobacter pylori: fasting the C57BL / 6 mouse obtained in the step 2, carrying out intra-gastric administration by helicobacter pylori, and feeding after 8 to 12 hours, wherein the inoculating in the step 3 is performed one time every other day, and performed 2 to 4 times in total; and 4, raising the mouse which has been inoculated for 2 to 4 times in the step 3. In the modeling method provided by the invention, the experimental time is short, the operation time is less, the order of magnitude of the helicobacter pylori infection is high, and the infection time is endurable; the model can be widely used for the method of helicobacter pylori infection animal in related researches.

The invention relates to a method for detection of P-glycoprotein (P-gp) mediated rhodamine 123 (Rh123) transport in a 3D type organ and an application thereof in screening of P-gp inhibitors. The method comprises the steps: firstly, sorting small intestinal crypts of C57BL / 6 mice, inoculating a petri dish containing a matrix gel, culturing in an Advanced DMEM / F12 culture medium, and forming the 3D type organ; and then carrying out qualitative and quantitative detection of P-gp mediated Rh123 transport and influence of the P-gp inhibitor verapamil on Rh123 transport by using the 3D type organ model. The method particularly comprises the steps of respectively co-incubating the 3D type organ with (1) the P-gp substrate Rh123 and with (2) the Rh123 and the verapamil, releasing the Rh123 in the 3D type organ by using an ultrasonic crushing method, and finally detecting the concentration of the Rh123 on a multifunctional microplate reader. The method has the advantages of being simple, rapid, and high in sensitivity, can be combined with the 3D type organ model for carrying out research of the P-gp mediated drug transport, and also can perform research of in-vitro screening of the P-gp inhibitors.

The invention discloses a method for constructing a tumor xenograft model in mice immunized with gastric cancer based on an organ-like method, and an application thereof. The invention relates to a medical tumor animal model, based on an organ-like method. For the first time, the present invention employs a quasi-organ technique, that is, using a microcarrier as substrate. The composite human gastric cancer cell line SGC-7901 / MKN-45 is transplanted subcutaneously into C57BL / 6 mice to establish a tumor model of human gastric cancer with normal immune function. The tumor model constructed by this method has good tumorigenicity, and the tumorigenicity rate is up to 75%. The HE staining and the immunohistochemistry of the transplanted tumor accord with the characteristics of human gastric cancer. Compared with the existing immunodeficient mice transplanted tumor model, the model can better study and further clarify the mechanism of tumor occurrence and development in the normal immune system, and also provides a more valuable new animal model for the research and development of anticancer drugs. The invention is applied to the tumor model field.

The invention relates to a construction method of a Membrane protein cDNA library and an application thereof. The method constructs a eukaryotic expression vector used for the specificity selection of the membrane protein. The expression vector can perform clone recombination with any cDNA clone library provided with the recombination site attL of Lambada bacteriophage so as to acquire a cDNA expression library. After the transfection of COS-1 cell, expression membrane protein clone is further selected by immunofluorescence so as to acquire the membrane protein cDNA expression library. As the application of the method, the liver lymphocyte membrane protein cDNA library of a C57BL / 6 rat is built up. The method is proved to be an effective proposal of the high flux specificity selection membrane protein through the identification and analysis of the library, and the method provides a powerful method to describe the map of the membrane protein of the cell surface of certain issue and to research the unknown membrane protein.

The present invention provides application of crosslinked polyethylenimine as an oncoprotein antigen vaccine vector, and overcomes the problem that protein antigen after penetration into the body can be degraded easily by enzyme, leading to weak immunogenicity of the vaccine. In an aqueous solution, the polymer material is capable of forming a complex antigen nanoparticle with antigen, in order to achieve loading of tumor protein antigen. The present invention uses RF33.70 as a target cell model, OVA as a model antigen, and C57BL / 6 mice bone marrow-derived dendritic cells as antigen cross presenting cells, and detects the antigen cross-presentation, cytotoxicity and in vivo anti-tumor effect of the complex nanoparticles formed by biodegradable PEI as the antigen vector and the OVA.

The invention relates to application of benserazide hydrochloride in preparing medicine for treating acute inflammation and belongs to the technical field of biological medicine. According to the application of benserazide hydrochloride in preparing medicine for treating acute inflammation, a treating effect of the benserazide hydrochloride on acute inflammation is researched through animal experiments, and experiment results show that the benserazide hydrochloride has an inhibiting effect on inflammatory factors generated by mouse acute inflammation caused by lipopolysaccharide. Specifically, a C57BL / 6 mouse acute inflammation model is induced to be constructed through the lipopolysaccharide, different concentrations of benserazide hydrochloride is utilized to treat the mouse, results show that the benserazide hydrochloride can reduce inflammatory factor level generated by C57BL / 6 mouse acute inflammation and pathological change degree of hepatic tissues, and the low dosage benserazide hydrochloride has the optimal effect. The benserazide hydrochloride is prepared into varieties of preparation required by clinic by adding carriers accepted by pharmacy, and the application of benserazide hydrochloride in preparing medicine for treating acute inflammation has the beneficial effect of providing a novel path for preparing acute inflammation medicine.

The invention belongs to the technical field of traditional Chinese medicines, and discloses a preparation method and application of a herba ardisiae japonicae alcohol extract. The preparation method comprises the following steps: after drying and crushing a herba ardisiae japonicae medicinal material at low temperature, weighing 0.1g of the crushed herba ardisiae japonicae medicinal material, and adding 0.1mL of methanol; after carrying out vortex oscillation, carrying out ultrasonic treatment and cooling; then weighing the weight, and supplementing the reduced weight with the methanol; uniformly shaking and filtering; taking subsequent filtrate and preserving for later use; weighing a bergenin reference substance, and adding the methanol to prepare a solution; determining the content of bergenin in the herba ardisiae japonicae alcohol extract by adopting an ultra-high performance liquid chromatography. According to the preparation method and the application of the herba ardisiae japonicae alcohol extract, the content of the bergenin in the herba ardisiae japonicae alcohol extract is 9.88mg / g and accords with quality standards; herba ardisiae japonicae can be used for reducing the quantity of lipid droplets in ST-2 and C3H10 cells which are obtained by inducing adipogenesis differentiation, and for inhibiting mRNA expression of adipogenesis differentiation characterization genes PPARgamma, C / EBPalpha and aP2; interventional treatment of the herba ardisiae japonicae can be used for effectively reducing the weight of C57BL / 6 mice fed by high fat diet.

The invention relates to construction for a number 1 chromosome substitution laboratory mouse strain C57BL / 6-Chr1NZW. According to the construction for the number 1 chromosome substitution laboratory mouse strain C57BL / 6-Chr1NZW, a new chromosome substitution laboratory mouse strain is constructed. A genetic background of the chromosome substitution laboratory mouse strain is based on a C57BL / 6 mouse, while wherein a number 1 chromosome is from an NZW mouse. According to the construction for the number 1 chromosome substitution laboratory mouse strain C57BL / 6-Chr1NZW, the chromosome substitution laboratory mouse strain can be used for research of character differences brought by different number 1 chromosome genomes under the same genome background, and can be a useful tool for research of the chromosome genome functions by adoption of a genetic method.

The invention relates to the field of experimental zoology, in particular to a pulmonary fibrosis animal model and a preparation method thereof, and application of chrysotile. Chrysotile fiber is used as an inducer and is instilled to SPF-level C57BL / 6 mouse by non-exposure process via intratracheal instillation, pulmonary fibrosis may be successfully induced in 21 days, operating is simple, model success rate is high, good conformity to pulmonary fibrosis process is achieved, post-forming stability is good, the fibrosis degree is irreversible and worsens gradually, self-healing tendency possible to a bleomycin model is overcome, and the model provides reliable animal model basis for the deep study on molecular mechanisms of pulmonary fibrosis.

The invention provides a construction method of a type I diabetes animal model. The method comprises the following steps: selecting C57BL / 6 mice, performing first tail vein injection, continuously performing intraperitoneal injection of a streptozotocin reagent for 5 days, performing ambrosia for 12 hours before injection, and allowing the mice to freely drink water. Two days after the injection procedure is completed, the ambrosia is carried out for 4 hours, and a glucometer is used for measuring blood glucose until the blood glucose value is 10 mmol / L or above. The statistical analysis of the blood glucose measurement result of the type I diabetes animal model shows that the C57BL / 6 mouse type I diabetes model constructed by using the streptozotocin is feasible and can be used for researching the diabetes pathogenesis and the anti-hyperglycemia intervention of medicaments and health-care foods. Compared with the prior art, the C57BL / 6 mouse type I diabetes model constructed by the invention has the advantages of high film forming rate, simple operation, no death of animals and good reproducibility.

The invention discloses an intervention effect of a ginsenoside Rg1 on autoimmune encephalomyelitis mouse, belonging to the field of Chinese herbal medicine monomer intervening disease. The autoimmune encephalomyelitis (EAE) model of cord sheath oligodendrocyte glycoprotein peptide MOG35-55 immune C57BL / 6 mouse is an international accepted multiple sclerosis mouse animal model, establishing the model, and simultaneously inducing, using the ginsenoside Rg1 to intervene, through observing the impact of the ginsenoside Rg1 on the phenotype and the histopathology of the EAE model, obtaining that the ginsenoside Rg1 can significantly reduce the incidence of the autoimmune encephalomyelitis. Therefore, the ginsenoside Rg1 has a certain role in the intervention of multiple sclerosis. The ginsenoside Rg1 can significantly reduce the disease incidence of the multiple sclerosis animal model-EAE mice.

The invention relates to a detection method for evaluating intestine toxicity of medicine by utilizing a 3D organ model. According to the detection method, firstly, small intestinal crypt of a C57BL / 6mouse is subjected to sorting, inoculated to a culture dish containing matrigel and cultured in an Advanced DMEM / F12 culture medium, and the shape of the 3D organ is observed. The 3D organ model is verified by utilizing a positive compound, and changes of the shape of the 3D organ and IC50 detection are included. Finally, the model is utilized for conducting toxicologic study on new medicine, andthe study particularly comprises the IC50 detection of the medicine, apoptosis conditions of the 3D organ and a mechanism of medicine induction organ apoptosis. The detection method has the advantages of being simple, quick and high in sensitivity, the 3D organ mode can be combined for conducting intestine toxicity detection on the medicine, and the medicine intestine toxicity mechanism can be conducted.

The invention relates to construction for a number 1 chromosome substitution laboratory mouse strain C57BL / 6-Chr1C3H / HeJ. According to the construction for the number 1 chromosome substitution laboratory mouse strain C57BL / 6-Chr1C3H / HeJ, a new chromosome substitution laboratory mouse strain is constructed. A genetic background of the chromosome substitution laboratory mouse strain is based on a C57BL / 6 mouse, while wherein a number 1 chromosome is from a C3H / HeJ mouse. According to the construction for the number 1 chromosome substitution laboratory mouse strain C57BL / 6-Chr1C3H / HeJ, the chromosome substitution laboratory mouse strain can be used for research of character differences brought by different number 1 chromosome genomes under the same genome background, and can be a useful tool for research of the chromosome genome functions by adoption of a genetic method.

The invention discloses a method for establishing an insulin resistance animal model. According to the method, feed with high content of selenium, sugar and fat and low-concentration fructose drinkingwater are used for inducing inbred line C57BL / 6 mice with highly similar genetic backgrounds in order to cause glycolipid metabolism disorder of the mice and then induce the occurrence of insulin resistance, and then the insulin resistance animal model is established. Through unique proportioning of the feed, a pathogenesis process of the insulin resistance caused by the change of a dietary structure is highly simulated, comparison between the induced animal model and a control group shows that animals in a model group are obviously different from animals in the normal control group in intakeglucose tolerance, insulin tolerance, animal insulin resistance index and insulin sensitivity index, and it is indicated that the animals in the model group have obvious insulin resistance. The method has the advantages that the special feed is adopted for feeding in two modes, adverse reactions caused by chemical reagents are eliminated, a molding method is simple, the molding rate is high, thecost is low, and the reproducibility is high.

The invention provides an oral squamous carcinoma cell strain as well as a preparation method and applications of the oral squamous carcinoma cell strain. The oral squamous carcinoma cell strain is derived from the MAL- / -C57BL / 6 mouse oral carcinoma tissue induced by drinking water containing 4-nitroquinoline-1 oxide. Through primary culture on the tumor tissue, the tumour cell is subjected to continuous subculture in vitro and grows lively. Through the monoclonal screening method, the oral squamous carcinoma cell strain is obtained, the cell strain can be cultured continuously in vitro and grows lively, and the following biological characteristics of the cell are analyzed: cell proliferation, growth curve, cell cycle and apoptosis, plate colony and migration, and the in-vivo tumor formation and pulmonary metastasis capacity of the nude mouse. The result proves that the oral squamous carcinoma cell strain has the typical biological characteristics of the tumor cell, can be taken as the cell model for researching the generation, development and metastasis mechanism of oral carcinoma, and also can be taken as a tool for screening or developing anti-cancer drugs.

The invention relates to the technical field of biological medicines, and relates to application of cyclic dinucleotide cGAMP combined PD1 antibody (Nivolumab) in preparation of antitumor combined drugs. Studies indicate that the cGAMP combined Nivolumab can significantly inhibit the growth of a variety of tumor cells, has obvious anti-tumor effect, and can be used for preparing the antitumor combined drugs. A C57BL / 6 mouse subcutaneous transplantation tumor model shows that the cGAMP combined Nivolumab has obvious inhibition effect on adenocarcinoma cell line MC38, lung cancer cell line LLC, breast cancer cell line EO771, melanoma cell line B16-F10 and colon cancer cells MC38, and the obvious inhibition effect is superior to single drug use effect, so that the cGAMP can be combined with the Nivolumab for the treatment of tumors.

The invention relates to construction for Chromosome 1 to replace wild mus musculus strain C57BL / 6. Zaozhuang 2-Chr1. A novel Chromosome C57BL / 6 replacing experiment musculus strain is constructed, a C57BL / 6 musculus is mainly used as the genetic background of the musculus strain, and a Zaozhuang 2 musculus is mainly used in the Chromosome 1. The Chromosome replacing strain can be used for studying character difference brought by different Chromosome 1 genomes under the background of the same genome, and a useful tool for studying the functions of the genomes by using a genetics method is achieved.

The invention discloses a building method of a mouse stomach cancer model. A four-week-old C57BL / 6 mouse is taken and placed in a tubular device, wherein movement of the mouse is restrained but the mouse can breathe freely in the tubular device; every stress period lasts for 25-30 days, and six to eight hours are needed every day; the interval between every two stress periods is 5-7 days, and two to three stress periods are needed; when stress is carried out, the mouse drinks water containing N-nitro compounds is provided for the mouse, medicine is fed for a week and is fed again after a week, normal drinking water is provided for the mouse at the medicine feeding intervals, and the mouse needs to be fed with medicine for five weeks; the mouse is bred for 35 weeks conventionally after the medicine feeding, and the mouse suffering the stomach cancer is obtained. According to prior literatures and reports, the stomach cancer formation rate obtained after MNU treatment is carried out on a C57BL / 6 mouse purely is about 30%; after the MNU treatment and the stress treatment are combined, the stomach cancer formation rate of the C57BL / 6 mouse increases to be 60%.

The invention relates to a recombinant adenovirus high-expression vector for promoting effective presentation of Hantavirus fusion protein G2S0.7. A G2S0.7-pCAG transfer vector containing G2S0.7 mosaic gene of Hantavirus 76-118 strain and a CAG promoter / enhancer is mainly transformed to promote the effective presentation of the high-expression Hantavirus fusion protein Hantavirus in an organism. By a gene recombination technology, the recombinant adenovirus transfer vector G2S0.7-pCAG containing the mosaic gene G2S0.7 is transformed, and Ub gene is connected to the vector. The recombinant transfer vector and the adenovirus vector are respectively subjected to double digestion, and a recombinant fragment Ub-G2S0.7-pCAG is integrated into the DNA of the adenovirus vector; and after packaging, purification and titer detection, rAd-Ub-G2S0.7-pCAG and recombinant adenovirus rAd-G2S0.7-pCAG containing the G2S0.7 mosaic gene and the CAG promoter / enhancer, which is constructed early by the inventor are respectively used for immunizing C57BL / 6 mice, immunological characteristics are researched, and results show that compared with the conventional recombinant adenovirus, the recombinant adenovirus can effectively improve partial humoral immune response level and partial cellular immune response level of organisms of experimental animals.

The invention belongs to the field of biological medicines and in particular relates to application of a farnesoid X receptor stimulant GW4064 in preparation of medicine for treating non-alcoholic fatty liver disease caused by senescence. A naturally senile C57BL / 6 mouse serves as a research object, triglyceride in the liver of the mouse is deposited with the increase of age, the mouse suffers from hypertriglyceridemia, and the change is independent from the influence of weight factors. The farnesoid X receptor expression is reduced in the liver of the senile mouse, the deposition of the triglyceride in the liver of the senile mouse can be inhibited by the farnesoid X receptor stimulant GW4064, the NAFLD disease caused by natural aging is treated, and a theoretical basis and experimental data can be provided for clinically treating the NAFLD disease.

The invention belongs to the field of nephrology, and relates to a chronic kidney disease angiosteosis experiment model and a making method thereof. The chronic kidney disease angiosteosis experiment model is established by using C57BL / 6 mouse molding through cooperation of combination of adenine and vitamin D with a high-phosphorus and low-protein diet intervention mode. The making method has the advantages of short modeling time and good stability; and the established chronic kidney disease angiosteosis experiment model can be further used to screen drugs for treating chronic kidney disease angiosteosis and research mechanisms of the drugs.