743 results about "Murine model" patented technology

The invention provides a strain of lactobacillus plantarum YS-AG23 with high antioxidant activity, and a rhizoma polygonati and radix angelicae sinensis fermented product obtained by fermenting rhizoma polygonati and radix angelicae sinensis with the strain of lactobacillus plantarum YS-AG23. In a D-galactose induced premature ovarian failure mouse model, the fermentation product can regulate up the levels of antioxidant and stress protection genes SOD2 and HO-1 and an anti-aging gene SIRT1 by regulating down the expression of a key gene AGER of an inflammation and oxidative stress pathway, so that the oxidative stress and inflammatory response of model mouse ovarian tissues are remarkably reduced; meanwhile, the serum hormone level can be specifically adjusted, the serum estradiol (E2) level can be increased, the follicle stimulating hormone (FSH) and luteinizing hormone (LH) levels can be reduced to be close to the normal range, the expression of anti-mullerian hormone (AMH) and hormone regulatory gene LHR of ovarian reserve markers can be up-regulated, the proliferative activity and estrogen secretion function of follicular granulosa cells can be improved, and the effect of preventing ovarian reserve is achieved. The wet weight of atrophic uterus and ovary of the model mouse is remarkably increased, uterine and ovary atrophy is effectively reversed, and the ovarian reserve function of the D-galactose induced premature ovarian failure model mouse is recovered. Besides, in an H2O2-induced human dermal fibroblast aging model, the yeast can improve the activity of aging cells in a concentration-dependent manner and reduce the level of reactive oxygen species (ROS) in the cells; in addition, by up-regulating the expression of a collagen synthesis key gene COL1A1 and down-regulating the expression of a cell senescence marker gene p16, skin cell senescence caused by oxidative stress can be effectively improved. In conclusion, the polygonatum sibiricum and angelica sinensis leavening can synchronously improve premature ovarian failure and skin cell senescence caused by oxidative stress by accurately regulating and controlling oxidative stress, inflammatory response and tissue function related genes, and has wide application potential.

The invention belongs to the technical field of biology, and particularly relates to a cryptosporidium parvum infection immunosuppression mouse model and a construction method thereof. Aiming at the problem that the development of cryptosporidiosis drugs and vaccines is seriously limited due to the fact that no model for successfully infecting mice with cryptosporidium parvum exists at present, the invention provides the construction method of the cryptosporidium parvum infection immunosuppression mouse model, which comprises the following steps: firstly, carrying out gavage treatment on the mice for 4-7 days by adopting cyclophosphamide; constructing to obtain an immunosuppressive mouse; feeding PBS (Phosphate Buffer Solution) containing cryptosporidium parvum oocysts through the mouth, and infecting for 7-21 days; and when oocysts are continuously detected from excrement of the mouse and the content of the oocysts is higher than the infection amount, constructing the cryptosporidium parvum infection immunosuppression mouse model. The cryptosporidium parvum infection immunosuppression mouse model is successfully constructed, can be used for subsequent drug and vaccine development tests for cryptosporidium parvum diseases, and has a good application prospect.

The invention relates to the technical field of image recognition, in particular to an adhesion focus positioning method for an endometriosis rat model, which comprises the following steps of: firstly, adjusting a clustering distance according to color difference between saturation connected domains and a position inertial system, and clustering to obtain a connected domain cluster; due to the fact that the connected domain clusters contain the ectopic glands, the clusters need to be screened to obtain the ectopic glands, the gland probability of each connected domain cluster is obtained according to the characteristic that columnar epithelial cells are distributed on the inner walls of the ectopic glands, and the possibility that each connected domain cluster belongs to the ectopic glands is judged; due to the fact that the ectopic glands are distributed in a concentrated mode, the abnormal probability of the connected domain clusters is obtained through the aggregation of the connected domain clusters with the high gland probability, the possibility that the connected domain clusters belong to the abnormal ectopic glands is further judged, and the focus positioning accuracy of the endometriosis rat model is higher.

The invention discloses a humanized osteoporosis mouse model and a construction method thereof, and belongs to the technical field of biotechnology and animal models. The invention aims at the core defects of large species difference, insufficient humanization degree, low pathological simulation degree, unstable phenotype and the like of the existing osteoporosis model. The humanized osteoporosis mouse model is constructed through five core steps of experimental animal pretreatment, collagen-nano hydroxyapatite bionic scaffold preparation and cell pre-implantation, immune-bone metabolism double-derived system construction, progressive osteoporosis induction and model identification and verification. According to the method, double-person origination of immune and bone metabolism systems is realized for the first time, the colonization rate and survival time of human cells are greatly improved, the pathological process of clinical postmenopausal osteoporosis is accurately simulated, the phenotype of the model is stable and uniform, the reactivity matching degree with clinical drugs is high, and the method can be widely applied to research and development of anti-osteoporosis drugs and research of pathological mechanisms.

The invention belongs to the technical field of biological medicines, and particularly relates to application of a small molecule compound P7C3 in preparation of a pharmaceutical preparation for inhibiting hepatic fibrosis. According to the invention, a CCl4-induced hepatic fibrosis mouse model is used for verifying the treatment effect of P7C3 on hepatic fibrosis. Meanwhile, the influence of P7C3 on HSC activation and proliferation is verified, eIF4A1 is a direct target of P7C3 for inhibiting HSC activation, and P7C3 is a promising anti-hepatic fibrosis therapeutic drug and an effective eIF4A1 inhibitor.

The invention discloses application of amantadine in inhibition of ITGAV in preparation of medicines for treating dry age-related macular degeneration, and belongs to the technical field of biological medicines. At present, no report for researching the ITGAV gene in the dry AMD exists, the specific action mechanism is not clear, and amantadine adaptation diseases do not include the dry AMD; according to the application disclosed by the invention, the gene target ITGAV is screened from mutual hair generation of microglial cells and RPE cells in a retina microenvironment, and amantadine is used for inhibiting ITGAV protein to relieve the progress of AMD, so that a new thought is provided for treating dry AMD. Experiments show that amantadine can inhibit ITGAV and delay EMT transformation of RPE cells. In a dry AMD mouse model induced by sodium iodate, RPE cell damage can be remarkably relieved by intraocular injection of amantadine, and the structure and function of a retina layer are improved. These results indicate that it may function in early intervention of dry AMD.

The invention discloses an application of lactobacillus mucosa LMSJ001 or a metabolite agmatine of the lactobacillus mucosa LMSJ001 in preparation of a medicine for improving intestinal aging, and the application research of the lactobacillus mucosa LMSJ001 in a constructed aging mouse model finds that the lactobacillus mucosa LMSJ001 can be used for preparing a medicine for improving intestinal aging. Therapeutic gavage and application of Lactobacillus mucosa LMSJ001 in colon organs for intervention can significantly improve aging indexes, alleviate histological aging performance, promote intestinal peristalsis of aged mice, reduce secretion of inflammatory factors of the aged mice, improve intestinal epithelial barrier functions of the aged mice and improve intestinal dryness of the aged mice. The lactobacillus mucosa LMSJ001 can generate a metabolite agmatine through arginine decarboxylase, and agmatine can improve related indexes of proliferation and differentiation of intestinal stem cells and reduce expression of related indexes of senescence. Based on the unique functions, the lactobacillus mucosa LMSJ001 or the metabolite agmatine of the lactobacillus mucosa LMSJ001 can be cooperated or complemented with other means, is used for preparing pharmaceutical compositions or detection kits and the like, and has very wide application prospects.

The invention belongs to the field of biological medicines, and particularly relates to application of loganic acid in preparation of a medicine for treating arthritis. Specifically, through experimental research of a collagen-induced arthritis mouse model, the loganin acid is found to be capable of remarkably relieving arthritis symptoms, relieving joint swelling, synovitis and cartilage injury in a dose-dependent manner, and reducing the levels of proinflammatory factors TNF-alpha and IL-6 in plasma. By further combining 16S rRNA sequencing analysis, the loganic acid can remarkably regulate the intestinal microbiota structure of arthritis mice, increase the abundance of Lactobacillus murinus in the intestinal tract and promote the generation of tryptophan metabolite indole-3-acetic acid, so that the effect of resisting rheumatoid arthritis is achieved through an intestinal flora dependent mechanism. Therefore, the invention provides a new application of loganin acid in preparation of a medicine for treating arthritis, and the compound has further research and application values as a potential small molecule medicine.

The invention belongs to the technical field of biological medicine, and particularly relates to application of corynebacterium casei in preparation of medicine for preventing and treating lung injury. By constructing a radon inhalation-induced lung injury mouse model, after exposure at different radon levels, the weight of a mouse is reduced, the lung coefficient is increased, the tidal volume of the lung, the ventilation volume per minute and the maximum inspiration flow are reduced, the respiratory function is inhibited, the alveolar cavity of the mouse is abnormally expanded, the pulmonary septum is thickened and fractured, and more fibrin exudation is accompanied; the traditional Chinese medicine composition can be used for effectively inhibiting pulmonary fibrosis and lung injury due to obvious inflammatory infiltration beside bronchial tubes, extravasated blood in small blood vessels and lung tissue collagen deposition, and can be used for gavage of corynebacterium casei to effectively inhibit the above phenomena. Therefore, the corynebacterium casei can be used for preventing and treating lung injury, especially lung injury caused by radon exposure.

The invention discloses an establishment method and application of a damp-heat intestine accumulation type ulcerative colitis animal model, and belongs to the field of animal models.The establishment method comprises the steps that S1, high-fat feed treatment is conducted, specifically, experimental animals freely eat high-fat feed for 6-10 weeks from the first day; s2, DSS treatment: feeding 1.5% (w / v) DSS solution to the experimental animal for 2 weeks; and S3, observing the damp-heat syndrome type and ulcerative colitis related indications of the experimental animals, and selecting the experimental animals meeting the scoring standard, thereby establishing the ulcerative colitis mouse model. Aiming at the situation that most patients induce ulcerative colitis due to excess energy due to the change of dietary structures and lifestyles at present, the invention constructs and evaluates the damp-heat accumulation type ulcerative colitis mouse model by inducing internal dampness by a direct pathogenesis (high fat diet), which is closer to the actual situation of clinical patients. Therefore, a model and reference are provided for research and treatment of pathology of the damp-heat accumulation type ulcerative colitis and pharmacological mechanisms of traditional Chinese medicines.

The application discloses a method for constructing a mouse model of atherosclerosis induced without high-fat diet, and relates to the technical field of disease model construction.The mouse model construction method provided by the application obtains a mouse model that spontaneously exhibits atherosclerosis symptoms without high-fat diet induction.The mouse model of atherosclerosis induced without high-fat diet can be used for researching the pathogenesis of atherosclerosis and screening and evaluating drugs for atherosclerosis.

The invention belongs to the field of biological genetic engineering, and particularly relates to a method for constructing a congenital heart disease mouse model. The invention provides gRNA (guide Ribonucleic Acid) for targeting a mouse Tmed1 gene. Nucleotide sequences of the gRNA are as shown in SEQ ID NO: 1 and SEQ ID NO: 2. Meanwhile, the invention provides a method for knocking out the mouse Tmed1 gene based on a CRISPR / Cas9 technology to obtain the mouse with the congenital heart disease. The Tmed1 mouse model is successfully constructed, a reliable mouse model is provided for congenital heart disease and visceral ectopic syndrome patients, subsequent detection and verification of upstream and downstream target gene functions and subsequent verification of development of related target drugs are facilitated, and great convenience is brought to disease mechanism research and drug research and development verification work.

The invention discloses a method for constructing a polycystic ovarian syndrome (PCOS) rat model with reduced sex hormone binding globulin (SHBG) on the basis of ruminococcus aureus with the preservation number of ATCC29149, and belongs to the technical field of animal models. According to the present invention, the bacterial strain is subjected to anaerobic culture to prepare the bacterial liquid, the dosage is 0.2-1 mL / 100 g, the intragastric administration frequency is one time per day to two times per week, and the time lasts for 4-8 weeks, such that the SHBG synthesis of the rat liver is induced to decrease, and the PCOS pathological characteristics such as testosterone level increase, estrus cycle disorder, ovarian polycystic change, insulin resistance and the like are initiated; the problem that a traditional hormone induction method does not conform to the physiological status is solved, the modeling period of a high-fat diet induction method is shortened, and an efficient experimental tool is provided for PCOS mechanism research and drug development.

The invention belongs to the technical field of vaccine adjuvants, and particularly relates to tubeimoside B liposome as well as a preparation method and application thereof. The preparation method comprises the following steps: dissolving phospholipid, cholesterol and PEGylated phospholipid in an organic solvent to form a mixed solution, and then evaporating the mixed solution to form a film; and hydrating the film by using an aqueous solution of tubeimoside B to finally prepare the tubeimoside B liposome. Compared with free tubeimoside B, the lipidosome has the advantages that the hemolysis percentage is obviously reduced, and antigen-specific humoral immune response and / or cellular immune response can be effectively enhanced when the lipidosome is used as a vaccine adjuvant. In an H1N1 influenza mouse model, the lipidosome combined antigen shows an excellent protection effect, which is shown in that the mouse weight fluctuation is relatively small and the lung virus load is remarkably reduced. The invention provides important theoretical basis and practical examples for developing low-toxicity and high-efficiency plant-derived vaccine adjuvants, and has a wide application prospect.

The invention relates to an N-acetyl-D-glucosamine composition with an anti-tumor effect and application of the N-acetyl-D-glucosamine composition, and relates to the technical field of medicines and health care products, the N-acetyl-D-glucosamine composition with the anti-tumor effect comprises the following raw materials by weight: 17-29 parts of N-acetyl-D-glucosamine and 5-10 parts of an extract of Tongshan. According to the invention, the N-acetyl-D-glucosamine and the extract in a proper proportion are matched for use for the first time, so that the synergistic interaction of the N-acetyl-D-glucosamine and the extract is realized. In-vitro animal anti-tumor experiments show that the N-acetyl-D-glucosamine composition provided by the invention can significantly reduce the tumor volume in a rat model, provides a new way for clinical treatment of tumor patients, and has a wide application prospect.

The invention belongs to the technical field of biological medicine, and provides CKS1B serving as an immunotherapy response prediction biomarker and application of the CKS1B, and according to the application, CKS1B serves as an immunotherapy response marker, and a CKS1B inhibitor is combined with an active component to treat a model mouse. In-vitro cell experiments are adopted to evaluate the immunotherapy prediction effect of the CKS1B as a biomarker on esophageal squamous carcinoma, a Cks1b overexpression tumor mouse model, a homologous mouse model and a human immune reconstruction mouse model are established, and a CKS1B inhibitor is combined with active ingredients to treat the two models; results show that the CKS1B inhibitor combined with the active component can promote removal of esophageal squamous carcinoma cells by CD8 + T cells, inhibit interferon signal channels and antigen presentation, effectively recover immune response, inhibit tumor cell proliferation and significantly reduce tumor volume, so as to achieve the purpose of treating esophageal squamous carcinoma.

The invention relates to the field of bioengineering, in particular to a bilateral cleft lip mouse model based on Pitx2-Cre conditional knockout of Bmp4 as well as a construction method and application of the bilateral cleft lip mouse model. The invention aims to construct a stable bilateral cleft lip mouse model. Experiments find that the Bmp4 gene is expressed in the epithelium of the frontal nose process and the maxillary process at the early lip stage in E9.25. The spatio-temporal expression mode of the Pitx2 gene in the early primordium epithelium is highly consistent with the spatio-temporal expression mode of the Bmp4 gene. Based on this, the inventor constructs a Bmp4 gene knockout model driven by an endogenous Pitx2 gene promoter, and finds that the mouse model shows a 100% bilateral cleft lip phenotype. Compared with the reported Netin-Cre, the Nestin-Cre has the advantages that the Nestin The bilateral cleft lip model constructed by the method is more stable, can better represent genetic diseases of human cleft lip, and has important research significance.

The invention discloses an application of a Beclin 1 targeting pharmaceutical composition in preparation of antitumor drugs. The pharmaceutical composition comprises stapled peptides targeting Beclin 1 and platinum chemotherapeutic drugs. The treatment effect of the pharmaceutical composition on ovarian cancer is verified through an in-vitro cell model and an in-vivo mouse model, the combined use of the stapled peptide i7-01s-31 targeting Beclin 1 and cis-platinum can further amplify the anti-tumor effect of the stapled peptide i7-01s-31 and cis-platinum, the synergistic interaction effect is achieved, and the pharmaceutical composition can be used for treating ovarian cancer. The problem that drug resistance is easily generated due to single use of cis-platinum in actual clinical application is avoided; meanwhile, the dosage of chemotherapeutic drugs can be reduced through drug combination, so that the treatment safety is improved, and the toxicity is reduced.

The invention provides an application of liquiritin in preparation of a PPAR gamma receptor partial agonist. Through a structure-based high-throughput virtual screening technology, it is found that liquiritin can be used as a partial agonist of a PPAR gamma receptor, and the relative activation efficiency of liquiritin is 35.9% of that of rosiglitazone; in-vitro experiments prove that liquiritin can remarkably promote glucose uptake and consumption of HepG2 cells and does not induce adipocyte differentiation; in-vivo experiments prove that liquiritin can effectively improve the blood glucose level of an insulin resistance mouse model induced by high fat diet and reduce the level of serum proinflammatory factors. Compared with thiazolidinedione compounds, liquiritin not only can relieve and treat insulin resistance or related metabolic diseases, but also does not cause adverse reactions such as weight gain, fat accumulation and myocardial hypertrophy, provides a new candidate compound for developing safe and effective anti-diabetic drugs, and has a wide application prospect.

The invention belongs to the technical field of animal model construction, and particularly relates to a non-alcoholic fatty liver disease mouse model based on humanized PNPLA3 I148M transgene as well as a construction method and application thereof, the construction method comprises the following steps: step i: designing CDS of a human gene and a key 3 'UTR element for patent medicine of a small nucleic acid drug in gene plasmid DNA of the constructed model; step ii, using Luc2 or iRFP to report gene expression in the gene plasmid DNA for constructing the model; and (iii) a disease model of the fatty liver disease induced by combining plasmid DNA transient transfer with high-fat diet. The mouse model constructed by the invention has the advantages that a specific exogenous gene expression signal of the liver is detected in real time through small animal living imaging, a human PNPLA3 protein is detected and overexpressed through serum ELISA, PNPLA3 I148M mRNA is detected and overexpressed through liver RT-qPCR, the phenotype of the non-alcoholic fatty liver disease of the liver is determined through 15-week-old pathology, and the whole process of modeling for 8 weeks does not need to be operated in a biosafety secondary laboratory.

The invention discloses a noninvasive chronic abacterial prostatitis rat model making device which comprises a multifunctional operation base, a lifting stand column, a rat posture adjusting mechanism, a rat fixing plate, a lifting support, a pitching angle adjusting mechanism and a rat prostate multi-mode stimulator. The lifting support is fixed on the base, the pitching angle adjusting mechanism is fixed on the lifting support, the rat prostate multi-mode stimulator is fixed on the pitching angle adjusting mechanism, the rat posture adjusting mechanism is fixed on the multifunctional operation base, and the rat fixing plate is fixed on the rat posture adjusting mechanism. According to the device, the chronic abacterial prostatitis rat model is constructed by combining prostate local thermal stimulation with prostate local high-frequency vibration stimulation, the prostate tissue of the rat is noninvasive in the whole modeling process, and the problem of modeling failure or low success rate caused by introduction of bacteria in the injection puncture process is avoided.

The invention discloses a construction and evaluation method of a PM2.5 induced chronic obstructive pulmonary disease acute exacerbation mouse model, and relates to the technical field of disease animal models. The invention aims to evaluate whether PM2.5 can induce acute exacerbation of COPD through a mouse model, a COPD model is established by adopting combined treatment of smoking and lipopolysaccharide (LPS), and an infectious AECOPD model is established by using common klebsiella pneumoniae (KP) nasal drop on the basis. Meanwhile, PM2.5 processing is added on the basis of the COPD model, and the effect of PM2.5 on acute exacerbation of COPD is evaluated. Through comparison with a classical model (an infectious AECOPD model), the potential effect of PM2.5 in induction of acute exacerbation of COPD is evaluated, and an important reference is provided for future research on an AECOPD animal model induced by air pollution.

The invention relates to a traditional Chinese medicine compound for preventing and treating colorectal cancer precancerous lesions and an animal model verification method. The traditional Chinese medicine compound comprises the following components in parts by weight: 25-35 parts of semen coicis, 8-10 parts of radix aconiti carmichaeli, 12-18 parts of herba patriniae, 10-15 parts of radix et rhizoma rhei, 2-4 parts of cortex moutan, 8-10 parts of semen persicae, 25-35 parts of wax gourd kernels and 8-10 parts of mirabilite. The traditional Chinese medicine compound is based on the classical prescription of the coix lacryma-jobi radix aconiti carmichaeli dahurian patrinia powder and is combined with the prescription of the radix et rhizoma rhei and peony soup, so that the defect that the coix lacryma-jobi radix aconiti carmichaeli dahurian patrinia powder is insufficient in blood stasis removing and stagnation eliminating effects is overcome; finally, a spontaneous colorectal cancer mouse model shows a good inhibition effect on colorectal cancer.

The present application discloses a gene point mutation mouse model construction method, comprising: by taking T274M mutation on Kcnq2 Exon6 as a target, designing sgRNA and ssODN; selecting a female mouse of a set week age as an ovum donor, injecting a certain dose of serum gonadotropin and human chorionic gonadotropin into the abdominal cavity of the female mouse, then mating the female mouse with a mating male mouse, and collecting zygotes of the mated female mouse; injecting Cas9 protein, ssODN and sgRNA into the pronuclei of the zygotes by means of a microinjection technique, transplanting the zygotes obtained after injection into the ampulla of the fallopian tube of a surrogate female mouse, and obtaining a neonatal mouse after the pregnant female mouse gives birth; respectively designing a pair of identification primers F1, R1 at two ends of a mutation site on the basis of a Kcnq2 gene point mutation site, identifying the neonatal mouse, and screening out a Kcnq2T274M / + point mutant mouse in which T274M mutation is successfully knocked.

The invention discloses a chronic obstructive pulmonary disease mouse model induced by long-term continuous cigarette smoke exposure as well as a construction method and application thereof, and the construction method comprises the following steps: introducing cigarette smoke with constant concentration into a smoke exposure device, so that a mouse is in a cigarette smoke exposure environment for a long time and maintains normal diet and drinking; performing macroscopic characterization and microcosmic index evaluation on the mouse model with the chronic obstructive pulmonary disease until clinical symptoms of the chronic obstructive pulmonary disease appear; wherein the smoke exposure device comprises a smoke mixing box, a sealed feeding exposure box and a control box, the sealed feeding exposure box provides a cigarette smoke exposure environment, and the control box maintains the stability of the cigarette smoke exposure environment. According to the method, the occurrence cause of the chronic obstructive pulmonary disease of the mouse is simulated by optimizing exposure parameter conditions, the clinical symptoms of the chronic obstructive pulmonary disease of the mouse model are evaluated in multiple dimensions, and the method accords with the symptoms and signs of the chronic obstructive pulmonary disease, is high in similarity and good in reliability, and can be used as a disease model for research of the chronic obstructive pulmonary disease.

The invention provides a pharmaceutical composition and application thereof in preparation of a compound preparation, and belongs to the field of medicines. The pharmaceutical composition comprises a potassium ion channel opener, a 5 alpha-reductase inhibitor and a phosphodiesterase 5 inhibitor. The pharmaceutical composition can synergistically promote treatment of androgen dependent diseases or prevention of complications, and by taking the pharmaceutical composition of minoxidil, finasteride and tadalafil as an example, the effects in a benign prostatic hyperplasia rat model and an androgenic alopecia mouse model are verified, which indicates that the minoxidil, finasteride and tadalafil have a synergistic interaction effect, and the pharmaceutical composition can be used for treating the androgen dependent diseases or preventing the complications. Good clinical application prospects are realized.

The invention belongs to the technical field of medicine, and particularly relates to an excrement exosome, an excrement exosome transplanting method and application thereof, and the excrement exosome is prepared through the steps of excrement collection, excrement dilution, excrement oscillation and centrifugation, excrement supernatant filtration, excrement exosome extraction, excrement exosome purification and the like. The obtained excrement exosome is high in purity and small in particle size, impurities and pollutants can be effectively removed in the extraction and separation process, the bioavailability in vivo is high, and the excrement exosome can be absorbed by cells so that the biological activity of the excrement exosome can be exerted. By acting on a colitis mouse model, it is proved that the colitis disease index can be remarkably reduced, the barrier function can be enhanced, and intestinal inflammation can be inhibited through excrement exosome transplantation; the nerve injury of the Parkinson mouse can be obviously improved. Therefore, the excrement exosome has the prospect of being applied to preparation of drugs for relieving intestinal inflammation, enhancing the intestinal barrier function and relieving nerve injury, and a new thought and direction are provided for treatment of intestinal and nerve injury diseases.

The invention belongs to the technical field of medical instruments, and discloses a surgical instrument for building a traumatic brain injury mouse model.The surgical instrument comprises a tube blade, a limiter, a holding piece and a vacuum device, the tube blade is used for rotationally cutting into brain tissue of a mouse, the limiter is movably installed on the tube blade in the axial direction of the tube blade, and the vacuum device is arranged on the holding piece; the limiting device is used for abutting against the skull of the mouse, the holding piece is provided with an airflow channel, one end of the holding piece is connected with the pipe blade, one end of the airflow channel is communicated with the pipe blade, and the vacuum device is connected to the other end of the holding piece and connected with the other end of the airflow channel. According to the surgical instrument for building the traumatic brain injury mouse model, the problems that an injury area is uncontrollable and a focus area is difficult to quantify and unify in the current traumatic brain injury mouse model building process can be solved, so that the success rate and efficiency of model building are improved to the maximum extent.

The invention relates to a gene point mutation mouse model construction method. The method comprises the following steps: designing sgRNA and ssODN by taking T274M mutation on Kcnq2Exon6 as a target spot; the method comprises the following steps: selecting a female mouse at a set week age as an ovum donor, injecting a certain dose of serum gonadotropin and human chorionic gonadotropin into the abdominal cavity of the female mouse, mating with a breeding male mouse, and collecting fertilized ova of the mated female mouse; injecting Cas9 protein, ssODN and sgRNA into the pronucleus of the fertilized ovum through a micro-injection technology, transplanting the fertilized ovum after injection into the oviduct pot belly of the female rat to be pregnant, and obtaining a newborn rat after the female rat to be pregnant is delivered; according to a Kcnq2 gene point mutation site, a pair of identification primers F1 and R1 are designed at the two ends of the mutation site respectively, and a newborn mouse is identified, so that a Kcnq2T274M / + point mutation mouse with T274M mutation successfully knocked in is screened out. The Kcnq2T274M / + point mutation mouse editing method is simple in preparation and high in repeatability, and a larger number of Kcnq2T274M / + point mutation mice can be obtained while the editing efficiency is improved.

The invention discloses application of an Hspa5 inhibitor in preparation of a medicine for preventing or treating anxiety disorder. An anxiety mouse model is constructed through chronic constraint stress (CRS), and in combination with medial amygdala kernel (MeA) transcriptome sequencing and qPCR verification, it is found that the endoplasmic reticulum molecular chaperone Hspa5 is remarkably up-regulated in the anxiety state. Furthermore, an Hspa5 specific inhibitor HA15 is locally injected into a MeA brain region, so that the anxiety-like behavior induced by the CRS is remarkably improved, and the exploration time of an open field experiment central region, the exploration time of an open arm of an elevated cross labyrinth and the exploration time of a bright box of a bright-dark box experiment are prolonged. The invention discloses the function of Hspa5 as a novel anti-anxiety target for the first time, and provides a direct experimental basis and a transformation direction for developing a novel anti-anxiety drug which is non-monoamine and targets an endoplasmic reticulum homeostasis.