144 results about "C57BL/6" patented technology

The present invention belongs to the technical field of biological pharmacy, and in particular relates to a preparation and purification method of tetra-hydroxyl xanthone mango aglycone (norathyriol), and an application thereof in hypoglycemic medicine. Based on biological experiments, mango glycoside is used as raw material; under the condition with phenol and hydroiodic acid, carbon-carbon bonds are broken to produce the crude product of mango aglycone; the crude product is separated and purified through silica gel column chromatography so that the mango aglycone is separated; the yield rate reaches 12 percent, the purity is higher than 95 percent, and the reproduction performance is good; the IGTT of C57BL/6 normal mice is effectively improved with the norathyriol; and simultaneously the quantity of insulin secretion induced by glucose in vivo for the C57BL/6 normal mice can be improved with the norathyriol. Therefore, the mango aglycone can be applied for preparation of hypoglycemic medicine.

The invention belongs to the technical field of biological pharmacy, and in particular relates to suppressive activity detection of tetrahydroxy norathyriol on protein-tyrosine-phosphatase (PTP) 1B, improvement of resistance function of insulin and accpication in medicament for PTP1B related diseases. Proved by in vitro enzyme suppression experiments, the invention testifies that mango aglycone is the competitive suppressant of restructuring humanized PTP1B with IC50 value being 9.2 mu m; mango aglycone can ensure the sensitivity of C57BL/6 normal mouse and ob/ob diabetes model mouse on insulin to be obviously improved, and reduce blood sugar of experimental animals, which suggests that mango aglycone is a novel suppressant of PTP1B, and can be taken as potential lead compound to be applied in preparing medicament for curing insulin resistance related diabetes, obesity and other metabolic syndrome, tumor and other PTP1B related diseases.

The invention relates to an escherichia coli anti-tumor targeted engineering strain, and a building method and application thereof. The escherichia coli anti-tumor targeted engineering strain, i.e., Escherichia coli Nissle 1917(Tum-5) is preserved in CCTCC (China Center for Type Culture Collection), and the preservation number is CCTCC NO:M 2017345. The anti-tumor targeted engineering strain contains an anti-tumor secretion express vector pET28a-Pvhb-SUMO-Tum 5; and the soluble expression and the secretory expression of anti-tumor blood vessels of tumstatin can be realized to generate an active region Tum-5. The invention also comprises the building method of the escherichia coli anti-tumor targeted engineering strain, and application of the escherichia coli anti-tumor targeted engineeringstrain to preparation of anti-tumor medicine. The tumor inhibiting rate of the escherichia coli anti-tumor targeted engineering strain on B16-bearing melanoma C57BL/6 mice reaches up to 52.95 percent.

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 discloses a method for establishing a green fluorescence BALB/c nude mouse model. The method includes: selecting female C57BL/6-Tg (CAG-EGFP) mice and male Foxn1nu mice to perform hybrid to acquire F1 child generation hairy fluorescence mice, selecting the female mice, selecting the female mice having sexual maturation and the parental generation male Foxn1nu mice to perform mating (backcross) to acquire an F2 child generation; generating segregation of characters of the F2 child generation, selecting female hairy fluorescence mice and the parental generation male Foxn1nu mice to perform backcross, and completing a backcross cycle; performing the backcross cycle for ten times to acquire an F10 child generation, and establishing the green fluorescence BALB/c nude mouse model. The invention also discloses application of an animal model established by the method to a brain tumor transplanting model. The method uses a homogenous introduction method to establish the new strain model mice which can stably express EGFP, and can lay a firm foundation for human glioma orthotopic transplantation tracking and research on glioma and a micro environment thereof.

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 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.

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 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 separation and culture method of testicular mesenchymal stem cells for expressing nidogen and an application of the testicular mesenchymal stem cells. The separation and culture method comprises the following steps: providing a nidogen-green fluorescent protein transgenic mice model adopting C57BL/6 as the genetic background; raising the mice for one or more than one week, separating the testis, removing envelopes and blood vessels, exposing the seminiferous tubule, then digesting by using collagenase IV, and filtering digested cell suspension; and sorting by a flow cytometer to obtain single cells and collecting the cells for expressing green fluorescent protein with the fluorescence intensity being 10 times or more than 10 times of the fluorescence intensity of negative control cells so as to separate the testicular mesenchymal stem cells for expressing the nidogen. The invention provides testicular mesenchymal stem/progenitor cells separated according to the preparation and culture method and self-renewed and proliferated cells obtained by culture in a culture medium and provides the application of the cells in preparing a composition for treating related diseases with low testosterone level.

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 present invention relates to the design and development of recombinant protein for the delivery of silencer RNA complex to mediate RNA interference since it represents a novel therapeutic approach to modulate several neurodegenerative disease-related genes across the blood-brain barrier (BBB). To overcome challenges due to this barrier for biologics and other biological complex, the present invention describes a method wherein peptide having sequence GGGGHLNILSTLWKYRC represented by SEQ ID NO. 9 known to target specific gangliosides was linked to a double-stranded RNA binding protein to bind and deliver silencer RNA to the brain parenchyma. The designed fusion protein comprising a double-stranded RNA-binding domain (dsRBD) of human Trans Activation response element (TAR) RNA Binding Protein (TARBP2) and a brain targeting peptide sequence that binds GM1. Conformation-specific binding of TARBP2 domain to silencer RNA results in the formation of homogenous serum-stable complex with GM1 targeting potential. Uptake of the complex in neural cells reveals selective requirement of GM1 for entry. Remarkably, the invention pertains to the systemic delivery of the complex comprising TARBP-BTP and silencer RNA in AβPP-PS1 mouse model of Alzheimer's disease (AD) led to distinctive localization primarily in the cerebral hemisphere in the hippocampus and brain cortex and in principle can work across other mammalian CNS targets. Further, the delivery of silencer-RNA mediated by brain targeting peptide fusion led to significant knockdown of BACE1, a therapeutic protease target in both AβPP-PS1 and wild type C57BL/6 mice. The invention establishes the emergent importance of fusion proteins in delivering therapeutic siRNA as a simple complex to brain tissues to treat neurodegenerative diseases besides Alzheimer's disease (AD). The complex is also useful to study gene function of hitherto unidentified genes/interplay of genes in mammalian systems and central nervous system.

The invention belongs to the technical field of animal model building and discloses a chronic food-borne Parkinson's disease mouse model and a building method and application thereof. The method includes: a 5% ferric citrate solution is used to continuous intervene a C57BL/6 mouse for 16 weeks, behavioral evaluation is performed through open-field, speed-up rotation and pole climbing experiments every other 4 weeks, and the mouse of the 16th week is subjected to a suspension experiment and a food award type Y-maze experiment; the measured mouse total journey, average speed, center times, center journey, center time and on-rod time of the mouse are evidently reduced as compared with those of the control, turning time and pole climbing time of the mouse are evidently increased as compared with those of the control, the results are time-dependent, and mouse sports coordination, balance ability, muscular tension and activity are lowered evidently. The chronic food-borne Parkinson's diseasemouse model and the building method have the advantages that the occurrence conditions of Parkinson's disease are simulated, and the chronic model is suitable for monitoring early-stage change and screening the early-stage diagnosis indexes of the Parkinson's disease; behavioral disorder can be quantified, and the pathological characteristic of the Parkinson's disease is dopaminergic neuron degeneration and loss characteristic lesion.

The invention provides a skin type lupus erythematosus mouse model and a construction method and application thereof. According to the skin type lupus erythematosus mouse model constructed by the construction method, pristane is combined with ultraviolet (UVB) stimulation to construct the skin type lupus erythematosus mouse model. A C57BL/6 mouse is continuously irradiated by a proper amount of UVB after being subjected to Pristane intradermal immune treatment. The skin type lupus erythematosus mouse model better simulates immune response generated by abnormal activation of mouse skin immune cells under UVB stimulation. The model can be used for discussing the occurrence mechanism of the disease progress of lupus erythematosus, researching and developing drugs for targeting lupus skin lesion and the like, and is of great significance in the aspect of accelerating and promoting scientific research and clinical transformation.

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 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 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.