43 results about "Animal Disease Models" patented technology

The invention relates to a recombinant adenovirus vector for efficiently inducing a pluripotent stem cell (PS cell) and method for inducing the PS cell by using the recombinant adenovirus vector. The recombinant adenovirus vector is characterized in that the fiber genes therein are B subgroup adenovirus fiber genes, and an Sox2 gene expression cassette and an Oct4 gene expression cassette are connected within the recombinant adenovirus vector in an operating way. The terminally differentiated cell or the adult stem cell of the mammal, particularly the human is infected in vitro, the Oct4 genes and the Sox2 genes are expressed in an ectopic way, and the terminally differentiated cell or the adult stem cell of the mammal, particularly the human can be induced into the PS cell efficiently and rapidly under the synergistic effect of adjusting the epigenetic-inheritance small-molecular medicament. The PS cell of the human can be used for the cell replacement therapy to treat diseases, and the PS cell of the mammal can be used for preparing the transgenic animal model and the animal disease model.

The invention relates to a composition and kit for quickly extracting circulating un-related karyocyte from peripheral blood of an animal disease model, as well as application thereof. The composition which is used for quickly acquiring circulating un-related karyocyte from mammal blood of an animal disease model (including a tumor animal model, a cardiovascular disease animal model and the like consists of buffer fluid, magnetic or non-magnetic immunomicrospheres coated by anti-related karyocyte surface marker antibody, and cell separation medium with certain density. Compared with commercial centrifugal medium which is universally applied currently on the market, the centrifuge time can be greatly shortened by applying the novel composition, and the cell recovery rate is stably increased to over 95 percent from the previous 60+/-20 percent. The erythrocyte is not required to be cracked, molten and broken by means of the method, so that the un-related circulating rare cells can be enriched and extracted from the peripheral blood in the animal disease model effectively and quickly.

The invention relates to a method for obtaining a lymphoma minipig disease model by knocking out P53 genes, and belongs to the field of medical animal disease models. The method mainly comprises the steps that TALEN target spots of the P53 genes are designed, P53 gene knockout TALEN plasmid pCAG-pP53-L and pCAG-pP53-R of the P53 genes are assembled, TALEN plasmid is transfected and screened to obtain a P53 gene knockout positive cell line, reconstructed embryos are constructed based on the somatic cell nuclear transplantation technology and transplanted into the body of a surrogate sow to continue to develop, and a piglet with the P53 genes knocked out is obtained; the function of the P53 genes is verified by detecting the expression level of RNA and protein of the P53 genes in tissue of the cloned piglet and downstream related genes, and lymphoma in the tissue of the piglet is identified through histopathology and an immunohistochemical method. The built lymphoma disease model has great significance in research and development of occurring, forming and medicine of human lymphoma, and the reliable model is provided for research and treatment of human lymphoma diseases.

The invention relates to a preparation method of a hepatocyte apoptosis animal disease model by coinduction of D-galactosamine and lipopolysaccharide, in particular to a simple inducer formula and a simple operation process which can generate a hepatocyte apoptosis rate above 80% and reproduce a stable and reliable hepatocyte apoptosis animal model. The invention also relates to the application of the hepatocyte apoptosis animal model to the research on the pathogenesis of hepatitis and evaluation of novel anti-hepatitis medicaments.

Disclosed are a cloned pig expressing green fluorescent protein (GFP) and a cloned pig having a 1,3-galactosyltransferase (GT) gene knocked out. Also, the present invention discloses methods of producing such cloned pigs, comprising the steps of establishing a somatic cell line; preparing a GFP-transfected or GT gene knock-out nuclear donor cell; producing a transgenic nuclear transfer embryo using the nuclear donor cell and a recipient oocyte; and transplanting the transgenic nuclear transfer embryo into a surrogate mother pig. The cloned pig expressing GFP of the present invention is useful for large-scale production of an animal disease model, and the GT gene knock-out cloned pig can be used as a organ donor allowing xenotransplantation in humans without hyperacute immune rejection.

The invention discloses a lipidosome microbubble metal-loaded-ICG self-assembly composite system which has a multimode imaging/sonodynamic therapy (SDT) function. The system shows sonodynamic performance better than pure ICG molecules and has sonodynamic therapeutic effects of fixed-point explosion of lipidosome microbubbles, drug fixed-point release and increase of animal disease models of tumor,infection and atherosis. The invention further relates to sonodynamic therapy guided by metal ion/ICG@Microbubbless ultrasonic imaging, fluorescent imaging and photoacoustic imaging. The system is simple to prepare, cheap in price and remarkable in therapeutic effect.

The invention discloses an establishment method and application of a novel rat hypertension model. The disclosed establishment method of the novel rat hypertension model includes the following steps that 1, a human-body intron-derived 27nt-microRNA sequence is provided to construct its high expression plasmids; 2, the high expression plasmids in the step 1 is mixed with an X-treme GENEHP DNA transfection reagent in a certain proportion, after dilution with normal saline, the reagent is injected instantaneously in a SD rat at high speed from the caudal vein of the rat, the blood pressure of therat is increased to a high level, and the novel rat hypertension model is formed. The rat hypertension model is an effective animal disease model, can be used for verifying the influence of human-body intron-derived 27nt-MicroRNA on the blood pressure and vasodilator factors of the normal rat and screening antihypertensive drugs, and has wide application prospects.

The treatment of various sensitive organs with low energy acoustic shockwaves has been proposed. However, the prior art is lacking in guidance as to what constitutes an efficacious minimum dosage or a safe maximum dosage for various target organs and tissues. Through extensive experimentation with cultured cells, live animals, and animal disease models, the inventors of the present disclosure have determined safe and efficacious shockwave energetic dosage ranges for vital and sensitive organs, including the brain, pancreas, kidneys, liver, and spleen, as well as for skin and subcutaneous tissues, peripheral nerves, and skeletal muscles.

The invention discloses a method for constructing a full-retina in-vitro culture model suffering from diabetes retinopathy and belongs to the field of animal disease models. The model construction method disclosed by the invention comprises the step of culturing a retina of a rat or fetal pig in a DMEM/F12 culture solution containing D-glucose, Neurobasal A, N2/B27 supplement, insulin, BSA and cpt-cAMP. According to the method disclosed by the invention, an in-vitro model with typical diabetes retinopathy symptoms can be rapidly obtained, consumed time is short, the method can be repeatable, and the application value is good.

The invention provides a method for establishing a mouse endometritis model. The method includes the steps that female mice of 8-12 weeks old are used as test animals, after the mice are subjected to general anesthesia, a mouse uterus perfusion apparatus is used for injecting 20 microliters of 2.5-5.0 mg/ml lipopolysaccharide (LPS) into each mouse uterus, and after 12-24 hours, it is judged that the model is successfully established by observing the infiltration condition of inflammatory cells in mouse uterus tissue slices and the content change of mouse uterus nitric oxide (NO) and myeloperoxidase (MPO). The invention further discloses the mouse uterus perfusion apparatus used for injecting drugs. The perfusion apparatus is simple in structure and reasonable in design, and medicine can be injected into mouse uteruses conveniently and rapidly. Compared with an existing method for establishing the endometritis model, the damage of operation techniques to experimental animal organisms is reduced, the model better approaches to a natural paroxysm, the interference with test results is small, the success rate is high, operation is simple, and an appropriate animal disease model is provided for diagnosis, treatment, prevention and control of endometritis.

Compositions for the in vivo delivery of a gene editing CRISPR/Cas9 complex was developed to eliminate integrated retroviral DNA sequences from latently infected human cells and animal disease models.

A non-human animal disease model for hepatitis B virus-associated liver disease is disclosed. The animal disease model is transduced with a hepatitis B virus genome in the liver cells thereof and exhibits the following symptoms: hepatitis B viral particles and hepatitis B viral DNA in the serum, hepatitis B virus (HBV) envelope and HBV e proteins in the serum, expression of HBV core and HBV envelope proteins in the liver but not in the kidney, heart, lung, brain, pancreas, spleen, stomach or intestine tissues. The animal disease model may develop hepatocellular carcinoma, exhibiting an elevated level of alanine aminotransferase as compared to a control animal without the hepatitis B virus genome in the liver cells thereof, and liver pathological symptoms such as tumor nodules, dysplasia, inflammatory infiltrates, necrosis and fibrosis.

The invention relates to a composition for constructing a urolithiasis cat disease model, and belongs to the technical field of animal disease model construction. Comprising the following raw materials in parts by volume: 40-60 parts of a pacifying agent, 5-15 parts of a stone inducing agent, 5-15 parts of a stone making agent and 25-35 parts of a nutritional agent. The method has the advantages that damage to an experimental animal body is small, self-recovery can be achieved in the later period, an ideal disease model is provided for exploring an effective treatment method, the method is suitable for being used in research of owl urolithiasis, and a model basis is provided for pathogenesis research of the disease and screening of treatment drugs. According to the animal model of the cat urolithiasis disease, which is obtained by the construction method disclosed by the invention, not only can the formation process of urinary calculi be deeply understood, but also the screening of anti-calculi Chinese and western medicines can be helped. The method has important significance in the aspects of discussing the pathogenesis of the urinary calculus, preventing and delaying the formation of the calculus, searching medicines for preventing and treating the urinary calculus and the like.

The invention provides a method for inducing lung injury models of experimental animals with different severity degrees by spraying modeling medicines with different dosages in trachea. The method comprises the following steps that an experimental animal is anesthetized, and after the animal reaches an anesthetized state, the animal is immediately held on a slope holding table in a supine mode sothat a tracheal opening can be found smoothly through the oral cavity of the animal; the mandible of the neck of the animal is illuminated by using a cold light source (or adopting a laryngoscope) soas to obtain a sufficient light source during subsequent intra-oral operation; the tongue of the animal is pulled out, the tongue root is pressed by a tongue depressor, and the trachea opening is found; an intra-tracheal atomizer is used, a needle-shaped head of the atomizer is directly inserted into the trachea of an animal to a certain degree (the size of the animal determines the insertion depth), the respiratory law of the animal is observed, modeling drugs of different doses are rapidly sprayed into the trachea at the moment that the animal inhales, and then the animal is kept on the slope fixing table for 5-30 seconds. The experimental animal lung injury model which is stable, controllable in death rate, better in consistency and capable of simulating clinical lung injuries of different severity degrees is obtained, and a more reliable animal disease model is provided for medical pre-clinical research.

The invention relates to a pre-clinical rapid experiment method of a drug. The method comprises the following steps: constructing an experiment animal disease model; feeding the drug to an experiment animal for the first time; after pre-set time, starting to carry out trace blood drawing and carrying out drug-time curve analysis on a blood sample; feeding the drug to the experiment animal for a plurality of times; observing a treatment effect on diseases and drug toxicity; after the drug effect is finished, feeding the drug to the experiment animal for the last time and carrying out the trace blood drawing again; analyzing an accumulation condition of the drug; dissecting the animal and carrying out mass spectrum scanning analysis on organ slices to obtain distribution data of the drug in each organ; proving pharmacology and toxicology according to the distribution data; and taking relative biological matrixes and carrying out bio-marker analysis. According to the pre-clinical rapid experiment method of the drug, provided by the invention, the pre-clinical experiment time can be greatly shortened and a drug development progress is accelerated; pharmacokinetic-pharmacological function-toxicology correlation analysis is relatively accurate; and injuries to the animal are reduced and the pre-clinical experiment cost is reduced.

The invention relates to application of conjugated linoleic acid glyceride and microcapsule powder thereof in preparing foods or medicines for preventing and treating high fat diet induced non-alcoholic fatty liver diseases. The invention discloses application of the conjugated linoleic acid glyceride or the microcapsule powder of the conjugated linoleic acid glyceride in preparing foods, healthcare products or medicines for preventing, relieving and treating high fat diet induced non-alcoholic fatty liver diseases. According to the application, CLA-TG (conjugated linoleic acid-triglyceride) is adopted to prevent and relieve non-alcoholic fatty liver diseases, and an animal disease model involved in the application is an HFD (high fat diet) induced non-alcoholic fatty liver disease model.The application has the effect characteristics that weights and fat to weight ratios are reduced, contents of blood ALT (alanine transaminase), AST (aspartate transaminase) and liver TG are reduced, and a liver lipid droplet accumulation phenomenon can be alleviated; and the application is good in effect, high in security and easy in product obtaining.

The invention belongs to the technical field of animal disease models, and specifically discloses an improved SD (Sprague Dawley) rat CD (Crohn's Disease) modeling method. The improved SD rat CD modeling method comprises the following steps: after fasting and narcotizing an SD rat, carrying out clysis by using a modeling drug (which is prepared by mixing 5 percent of a TNBS (Trinitro-Benzene-Sulfonic Acid) water solution with absolute ethyl alcohol according to a volume ratio of 1 to 1 and is 0.2 ml/100 g in body mass) according to the dosage of 50 mg/kg, adopting a body position of enabling the head of the SD rat to face downwards and the tail to face upwards after leading in the modeling drug so as to prevent clysis liquid from flowing outwards, and reversely carrying the SD rat for 1 to 2 minutes. The improved SD rat CD modeling method disclosed by the invention is simple and convenient in operation and good in repeatability, and a model accords with the features such as diarrhea, bloody stools, intestinal obstruction, weight lose, colonic ulcers and adhesion and abscess between intestinal canals and between an intestinal canal and visceral organs or tissues of clinic human CD; the modeling success rate is high, and the animal death rate is low.

The invention provides an animal disease model capable of accurately positioning thoraco-abdominal aortic aneurysm. According to a method, aorta ligation is used, and the thoraco-abdominal aortic aneurysm model capable of achieving accurate positioning is built, so that the aortic constriction degree is higher on the basis of thoracic aorta constriction, and the ligation position is more flexibleand is located on the arcus aortae part or the subhepatic aorta abdominalis part between the right innominate artery and the left common carotid artery; changes of high pressure and blood flow speed at the ligation position are used for inducing aortectasis, then the aorta structure and the hemodynamics are changed, the aortic aneurysm is caused, and observation can be conveniently achieved through modern medical means such as Doppler color ultrasound and section.

The invention provides a construction method of a persistent gouty arthritis animal disease model. The method comprises the following steps: embedding an MSU crystal in an articular cavity for 3-5 times at an interval of 3-5 days; and before an MSU crystal is embedded in an articular cavity, damaging femoral intercondylar fossa cartilage to cause cartilage defects. According to the method, the MSU embedding mode and frequency are redesigned, the pathophysiological process of recurrent gout attack is simulated by adopting repeated intervention measures of sufficient MSU, the operation is simple and repeatable, complications such as incision-related infection and MSU leakage are avoided, a producible, economical and accurate model is provided for in-vivo research of MSU deposition and local structure erosion in gouty arthritis, and a good research platform is further provided for research of related diseases.

The invention discloses a method for building a disease model for mice infected with bacterial sepsis and judging disease situations and particularly, escherichia coli is adopted for building a disease model of mouse bacteremia. According to the method, through building a relationship between observed symptoms and blood procalcitonin in human body, a mouse bacteremia disease development degree isfirstly innovatively and scientifically divided into six grades, so that the disease development degree of a bacteremia animal disease model is conveniently judged, and a better model building basis is laid for the development of diseases, screening of treatment drugs and scientific evaluation of treatment effects. By adopting the method, the severity degree and the treatment condition of a disease in a mouse can be effectively, scientifically, rapidly and accurately judged; the defects of only taking a death rate as a model building index and insufficiently sensitive and accurate judgment ofthe treatment effects of the screened drugs can be made up better; and the model building method provides a convenient, rapid and accurate judgment method for successfully building a drug screening model for treating bacteremia.

The invention discloses a CRISPR/Cas9 system and application of the CRISPR/Cas9 system in construction of swine-derived recombinant cells for resisting amyotrophy protein gene defects. The invention provides an sgRNA combination, the sgRNA combination is composed of sgRNADMD-Ug3 (the target sequence binding region is shown as the 1-20th nucleotide in SEQ ID NO: 8) and sgRNADMD-Dg3 (the target sequence binding region is shown as the 1-20th nucleotide in SEQ ID NO: 11). The invention provides a kit, the kit is composed of a plasmid pKG-U6gRNA (DMD-Ug3) of sgRNADMD-Ug3 obtained through transcription, a plasmid pKGU6gRNA (DMD-Dg3) of sgRNADMD-Ug3 obtained through transcription, and a plasmid pKG-GE3. The sgRNA combination or the kit can be used for preparing recombinant cells or preparing a Duchenne muscular dystrophy animal model. The recombinant cells are anti-amyotrophy protein gene defect cells and can be used for preparing an animal disease model through somatic cell cloning. A solidfoundation is laid for the preparation of a Duchenne muscular dystrophy swine model, and has important application value for the research and development of Duchenne muscular dystrophy medicines.

The invention belongs to the technical field of animal disease model preparation, and relates to a device for preparing an irritability gastric mucosal lesion animal model. The device mainly consists of a constant-temperature water bath kettle, a mouse cage lifter, a mouse cage, a heating rod, a temperature sensor, a control circuit, a control circuit working power supply, a heating rod power supply switch and a heating rod power supply, wherein a water tank is arranged in the shell of the water bath kettle; the mouse cage lifter, the heating rod and the temperature sensor are also installed in the shell of the water bath kettle; one side of the shell of the water bath kettle is provided with an electrical cabinet; the control circuit, the control circuit working power supply, the heating rod power supply switch and the heating rod power supply switch are all arranged in the electrical cabinet. A use result indicates that the device has the advantages of simple structure, convenience in use, good effect and reasonable construction, the experiment animal water spreading depth can be flexibly controlled, the molding success rate is improved, the stability degree in the experiment animal molding process can be strictly controlled, and the molding difference caused by an external factor is reduced.