38 results about "Miniature swine" patented technology

The invention provides a porcine reproductive and respiratory syndrome virus receptor CD163 knock-out swine and a cultivation method thereof. A constructed CD163 gene knock-out carrier pSSC-Larm-Sarm-CD163 uses a genome of Chinese experimental miniature swines as a template, and adopts a PCR method to amplify a homologous left arm and a homologous right arm of the swine CD163 gene; the cloned homologous left arm and the homologous right arm of the CD163 gene are subcloned to a pSSC-9 carrier respectively; and the porcine reproductive and respiratory syndrome virus receptor CD163 knock-out swine is cultivated, wherein the CD163 gene is not expressed, and the swine is not infected by the porcine reproductive and respiratory syndrome virus.

The invention provides the labeled molecule and the method for identifying wu zhishann miniature pig with labeled molecule. The labeled molecule possesses the nucleic acid sequence 1 in sequence table. The method comprises the following steps: using the DNA of pig as mold, making the 2 nucleic acid sequence and 3 nucleic acid sequences carry out PCR augmentation, and checking that if there is 88bp strip in augmentation product. The invention lays the foundation of utilizing wu zhishann miniature pig inbred line.

The invention relates to the biological cell technology field, particularly a method for separating and culturing miniature swine bone marrow-derived endothelial progenitor cells. The conventional EPCs culture method has the disadvantages of complexity and greatly reduced cell harvest rate by using immunomagnetic bead separation. The method in the invention comprises the steps of directly using myelomonocyte to conduct culture in vitro to obtain EPCs, adding growth factors in culture solution, planting in a definite density in a culture flask / vessel or a glass sheet enveloped by fibronectin, and conducing appropriate culture to obtain EPC. The method with high repeatability simplifies the steps of the conventional culture method, effectively avoids unnecessary cell loss caused by the conventional separation and culture method, increases the pick-up rate of EPCs, and saves a large amount of funds. The method of the invention also prepares for clinical application in future, and can reduce the usage of bone marrow and provide a technology platform for curing traumatic or ischemic diseases by autotransfusion.

The invention provides a targeting vector with the swine ApoE gene knocked out as well as a construction method of the targeting vector. The targeting vector takes the carrier pLoxPneo as a skeleton carrier, and comprises a positive screening component, a negative screening marker gene and the 5' homologous arm and 3' homologous arm of the swine ApoE gene. The invention further provides applications of the targeting vector in preparing animal models with the swine ApoE gene knocked out. With the adoption of the method provided by the invention, the healthy homozygote miniature pig with the ApoE gene knocked out is successfully obtained, therefore, a foundation is laid for the development of the novel medicines for treating cardiovascular diseases, and meanwhile, a guarantee is provided for developing pathological study, drug screening, efficacy evaluation and the like.

The invention discloses a method for assisting the judgment of a Wuzhishan minipig inbred line. The method for assisting the judgment of the Wuzhishan minipig inbred line and / or a non-Wuzhishan-mountain minipig inbred line, which is provided by the invention, detects that the 1400th position of deoxyribonucleotide from the 5' end of GenBankAccession Number DQ406743 of a pig to be detected is C or T, and determines that the genotype of the pig to be detected is CC, CT or TT; the pig to be detected with the CC genotype is an alternate Wuzhishan minipig inbred line; and a pig with the CT genotype and a pig to be detected with the TT genotype are non-Wuzhishan-minipig inbred lines. The method can be applied to the seed breeding of the Wuzhishan minipig inbred line. The method can firstly carry out preliminary screening on all the pigs in a swinery to be detected, eliminates the non-Wuzhishan-minipig inbred line, finds out the alternate Wuzhishan minipig inbred line and combines with other methods to carry out further validation. The invention can also be used for detecting whether the Wuzhishan minipig inbred line purchased in the market is counterfeit or not.

Miniature swine whose genomes contain sequences characteristic of pig endogenous retrovirus genes but which are non-infectious to humans are disclosed as sources of organs, tissues and cells for introduction into human recipients afflicted with diseases, or at risk of diseases, whose etiology involves the presence of inadequately functioning organs and for which xenotransplantation of such organs, tissues and cells would have a palliative effect. Methods of producing such animals and for screening animals for the desired properties are also disclosed.

The present invention discloses an experimental method for evaluating eye irritation by using miniature pig erythrocytes. The method mainly comprises: (1) preparing a suspension of miniature pig erythrocytes; (2) carrying out a miniature pig erythrocyte hemolysis test; (3) carrying out a protein denaturation experiment; (4) predicting a model; (5) carrying out classification and judgment. According to the present invention, the source of the erythrocytes is reliable and safe; the tested material is easy to obtain, and has good uniformity; the test system is simple and cheap, and no special equipment is required; the detection method is rapid, sensitive and universal; the method of the present invention can be directly applicable for safety evaluations of chemicals, cosmetics, pesticides, eye medicines and other substances possibly contacting the eyes, wherein the chemicals comprise surfactants, disinfection products, detergent and the like, the cosmetics comprise shampoo, bath foam, eye cream and the like; the method further can be applicable for rapidly screening, classifying and identifying the eye irritations of the raw materials, the formulas or the products.

The invention discloses a real-time fluorescence quantitative PCR detection method of campylobacter jejuni and a kit. In the invention, a primer and a Taqman probe are designed according to a conserved region of a campylobacter jejuni flagellin A (flaA) gene for qualitatively detecting the copy number of nucleic acid of the campylobacter jejuni in a fecal sample of a clinical patient, the fecal samples of a monkey and a dog, and the sample of ileocecal contents of a miniature swine, an upstream primer sequence SEQ ID NO:1, a downstream primer sequence SEQ ID NO:2 and a Taqman probe sequence SEQ ID NO:3. The invention has a great practical significance on quality control of relevant people and animal derived biological products and the inspection and quarantine field of import and export animals, and can ensure the quality of the relevant biological products. The detection method and the kit of the invention can be applied to diagnosis of campylobacter diseases and verification of the campylobacter jejuni in the biological products, thus having wide application prospect.

The invention discloses a preparing method for a small-sized pig 2-type diabetes model. The method comprises the steps of feeding high-fat high-sugar daily feed to a small-sized pig, continuously feeding for 280-320 days, wherein the high-fat high-sugar daily feed comprises the following components of 55-65 parts of complete feed, 25-35 parts of sugar and 5-15 parts of animal source fat. The small-sized pig 2-type diabetes obtained by the preparing method can be used for screening medicines for treating the 2-type diabetes model. The method is simple in step, convenient to operate and lower in cost, can be used for inducing the small-sized pig to present the impaired glucose tolerance and insulin resistance, the clinical features of the 2-type diabetes occur, a modeling success judgment standard is determined, the repeatability is strong, and the obtained small-sized pig 2-type diabetes model can be used for researching and screening the medicines for treating the 2-type diabetes model and has wide application prospect.

The invention discloses a preparation method of a miniature pig with a human complement regulatory protein knocked into a Rosa26 site at a fixed point, which is characterized in that a human complement regulatory protein gene is inserted into a pig ROSA26 gene site in a fixed-point gene recombination mode, so that the miniature pig with constitutive expression of the human complement regulatory protein is constructed. The constructed miniature pig not only avoids the unclear genetic background caused by exogenous random insertion and the unstable expression quantity caused by chromosome separation in the progeny breeding process, but also provides important animal resources for the research of xenotransplantation and immune related diseases in the future, and has important clinical application value.

The invention relates to a method for constructing an acute kidney injury (AKI) model of miniature pigs. Cisplatin at a dosage of 3.8-4.0 mg / kg is adopted for the first time, the male Ba-ma mini-pigsare selected as objects for constructing the model, and the acute kidney injury (AKI) model of the miniature pigs is constructed successfully in a mode of opisthotic intravenous injection. The acute kidney injury (AKI) model of the miniature pigs is constructed successfully for the first time, the problem is solved that an rodent model in the prior art has large differences from human acute kidneyinjuries in genetics, kidney anatomy morphology, physiology, administration modes, pathogenic mechanisms and other aspects, a closer and more reliable animal model for preclinical research of drugs is provided for study of the human acute kidney injuries, and a basis is provided for earlier prevention and treatment of cisplatin-AKI in clinical patients.

The invention relates to inbred line miniature pig auxiliary hind limbs, and belongs to the technical field of pig breeding. The inbred line miniature pig auxiliary hind limbs comprise a fixing plate,a connecting belt and a supporting rod; and the fixing plate is provided with a connecting belt used for connecting the hind limbs of a pig, and the lower end of the fixing plate is provided with a supporting rod capable of rotating by a certain angle. The fixing plate is firmly connected to the hind limbs of the inbred line miniature pig through the connecting belt, then the fixing plate is supported through the supporting rod, stable auxiliary supporting is provided for the pig from the side face of the hind limbs, the hind limb bearing of the pig is assisted, the normal function of the hind limbs of the inbred line miniature pig is recovered, and movement, ingestion, drinking and mating of the inbred line miniature pig are assisted; and meanwhile, in the advancing process of the pig, the supporting rod can rotate and move along with the shanks of the hind limbs, so that the pig walks more naturally and smoothly.

The invention discloses a modeling method of a Bama miniature pig excess heat syndrome model. A Bama miniature pig is taken as an experimental animal to manufacture the excess heat syndrome model by intramuscular injection of endotoxin. The method is simple to operate, high in modeling success rate and easy to popularize and apply, the animal model of the excess heat syndrome of the traditional Chinese medicine can be improved, objective reference standards are provided for syndrome differentiation and treatment and quantitative identification of the syndrome, and the modernization development of the animal model of the syndrome of the traditional Chinese medicine is promoted.

The invention discloses a special functional feed additive for experimental miniature pigs. The special functional feed additive is prepared from the following components in parts by weight: 8-12 parts of a Chinese herbal medicine extract, 1-3 parts of beta-nicotinamide adenine dinucleotide, 6-10 parts of fish protein amino acid, 5-10 parts of plant polysaccharide, 3-6 parts of soybean meal, 1-3 parts of multivitamins, 0.3-0.6 part of table salt, 0.5-1 part of taurine and 20-30 parts of rice bran. The special functional feed additive for the experimental miniature pigs is small in mixing amount, can effectively improve the immunity and digestibility of the organism, is balanced in nutrition, and has the effects of regulating the organism function and mental state of the experimental miniature pigs, relieving stress injury, promoting the repair of the organism after stress and improving the adaptability and the anti-stress capability; the health and experimental application value of the experimental miniature pigs can be ensured, the loss caused by various stresses in the production and experimental application process of the experimental miniature pigs is effectively reduced, and the animal welfare is guaranteed.

The invention discloses a special protection and fixation cage for miniature pigs. The special protection and fixation cage comprises a cage body (1) and a base (2); the cage body (1) is fixed at the top of the base (2); the cage body (1) is of a grating type structure; a cage door (3) is arranged on the cage body (1); a grating supporting plate (4) is arranged in the cage body (1); a lifting device (5) is arranged in the base (2); and the grating supporting plate (4) can move in the cage body (1) in the vertical direction under the action of the lifting device (5). According to the special protection and fixation cage for the miniature pigs disclosed by the invention, the miniature pigs can be effectively fixed and cannot move; and therefore, the success rate of technical operation, such as anesthesia, blood sampling and puncture, is increased.

The invention relates to an underwater breathing support system for Bama miniature pigs. The underwater breathing support system comprises a high-pressure gas cylinder (13), a secondary pressure reducing device (14) and a breathing mask (15), wherein the high-pressure gas cylinder (13), the secondary pressure reducing device (14) and the breathing mask (15) are connected through ventilation connecting members. According to the invention, the underwater breathing support system can be repeatedly used, 100% sealing can be achieved in cooperation of an adhesive tape and a glass glue, the underwater breathing of Bama miniature pigs is fully guaranteed, drowning and inhaling are avoided, and subsequent test failures are avoided.

The invention relates to an experimental miniature pig feeding fence, and belongs to the technical field of experimental miniature pig breeding. The experimental miniature pig feeding fence comprises a supporting frame, a feces leaking plate, a feces tray and barriers, wherein the bottom of the supporting frame is connected to the feces leaking plate in a clamped mode, feces leaking holes are evenly formed in the feces leaking plate at intervals, the feces tray located below the feces leaking plate is mounted on the supporting frame in a sliding mode, the fence is fixedly mounted on the periphery of the supporting frame, vertical sliding rods are correspondingly mounted in the middle of the front side and the middle of the rear side of the supporting frame, and sliding grooves matched with the sliding rods are formed in the front ends and the rear ends of the barriers correspondingly. According to the experimental miniature pig feeding fence, the barriers convenient to disassemble and assemble are arranged in the middle of the feeding fence, the barriers are disassembled or assembled according to the feeding scale, the pig feeding space is reduced or enlarged in applicability, the feeding space can be saved to the maximum extent, the feeding cost can be reduced, meanwhile, feces of pigs in the feeding fence can be rapidly discharged by arranging the feces leakage plate and the feces disc, the clean feeding environment is guaranteed, the miniature pigs can grow healthily, and breeding, popularization and utilization of clean-level and SPF-level experimental miniature pigs are accelerated.

The invention discloses a specific microsatellite primer composition for identifying experimental minipig varieties and a method for identifying experimental minipig varieties. The primer composition includes 3 pairs of primers. The present invention is based on the microsatellite sequence in the genome of the miniature pig breed, and obtains the specific microsatellite sequence and primers among different breeds of miniature pigs for experiments through population test verification and screening, optimizes the test conditions and usage methods of the primers, and solves the problem of Identification of miniature pig breeds for experimental use.

The invention discloses a specific microsatellite primer composition used for identifying an experimental miniature pig variety and a method for identifying the experimental miniature pig variety. Theprimer composition comprises three pairs of primers. According to the invention, on the basis of microsatellite sequences in the genome of the miniature pig variety, the specific microsatellite sequences and primers among different experimental miniature pig varieties are obtained through verification and screening of population tests; test conditions and use methods of the primers are optimized;and identification of the experimental miniature pig variety is solved.

The invention discloses a diabetic pig chronic skin ulcer model construction method, and relates to the technical field of skin ulcers. The method comprises the following specific steps: constructing a Bama miniature pig diabetes model, namely constructing the diabetes model in a Bama miniature pig body; 3D printing foreign matter filling design is conducted, a wound is made on the pig epidermis, and the wound is filled with foreign matter; and constructing a chronic skin ulcer model of diabetes, wherein the chronic skin ulcer model is constructed in the pig wound. The method is suitable for large animal models with relatively thick skin tissues, especially pigs; model formation is evaluated from multiple angles, and formation of diabetic chronic skin ulcers is better simulated from pathogenic factors, pathological changes and mechanism changes.

The invention relates to an underwater breathing mask for Bama miniature pigs. The underwater breathing mask comprises a silica gel sleeve (1), a hard shell (2), a first circular supporting plate (3),a second circular supporting plate (7), a plurality of ventilation copper pipes (4) and a plurality of silica gel pads (6); the silica gel sleeve (1) is of a straight cylindrical structure with an opening in one end, and the opening is in a horn shape; the silica gel sleeve (1) is divided into a first rubber sleeve section (8) and a second rubber sleeve section (9); the first rubber sleeve section (8) is nested in the hard shell (2), and a first circular supporting plate (3) and a second circular supporting plate (7) are correspondingly arranged on the outer side and the inner side of the first rubber sleeve section (8) respectively; wherein the hard shell (2) is of a straight cylindrical structure with an opening at one end; and the two ventilation copper pipes (4) are respectively and correspondingly inserted inwards from the round holes on the hard shell (2), and the silica gel pads (6) are arranged between the two ventilation copper pipes (4).

The invention provides a miniature pig tracheal intubation auxiliary tool, which comprises two supporting columns and an opening device arranged between the two supporting columns, the opening device comprises an upper mounting plate and a lower mounting plate, and a fixing frame arranged on the mounting plates and used for supporting the oral cavity of a miniature pig; the supporting column is provided with a driving device which is connected with the upper mounting plate and the lower mounting plate so as to drive the two mounting plates to be close to or away from each other, and the lower mounting plate is detachably connected with a catheter fixing device used for fixing an intubation tube. The fixing frame of the two mounting plates in the opening device between the two supporting columns are used for supporting the upper jaw and the lower jaw of the miniature pig respectively, the two mounting plates are far away from each other through the driving device, and therefore the oral cavity of the miniature pig is opened, sufficient operation space is provided for experimenters to conduct tracheal intubation, the working intensity of the experimenters is relieved, moreover, the intubation operation can be completed by only one person without assistance of multiple persons, the structure is simple, the use is convenient, and the intubation accuracy is improved.

The present invention discloses an experimental method for evaluating eye irritation by using miniature pig erythrocytes. The method mainly comprises: (1) preparing a suspension of miniature pig erythrocytes; (2) carrying out a miniature pig erythrocyte hemolysis test; (3) carrying out a protein denaturation experiment; (4) predicting a model; (5) carrying out classification and judgment. According to the present invention, the source of the erythrocytes is reliable and safe; the tested material is easy to obtain, and has good uniformity; the test system is simple and cheap, and no special equipment is required; the detection method is rapid, sensitive and universal; the method of the present invention can be directly applicable for safety evaluations of chemicals, cosmetics, pesticides, eye medicines and other substances possibly contacting the eyes, wherein the chemicals comprise surfactants, disinfection products, detergent and the like, the cosmetics comprise shampoo, bath foam, eye cream and the like; the method further can be applicable for rapidly screening, classifying and identifying the eye irritations of the raw materials, the formulas or the products.

The invention discloses a method for preparing a human complement regulatory protein miniature pig at the Rosa26 site. The invention inserts the human complement regulatory protein gene into the pig ROSA26 gene site through site-directed gene recombination, thereby constructing The minipigs constitutively expressing human complement regulatory proteins were obtained. The constructed minipigs not only avoided the unclear genetic background caused by the random insertion of foreign sources and the unstable expression level caused by the segregation of chromosomes during the breeding process of offspring, but also laid a solid foundation for future development. The study of xenotransplantation and immune-related diseases provides important animal resources and has important clinical application value.

The invention discloses an SNP marker related to miniature pig type 2 diabetes and an SNP marker detection method as well as an application of the SNP marker. The SNP marker is G / A polymorphism located at a part 325bp of an exon No.1 of a Leptin gene, and the nucleotide sequence of the Leptin gene with the SNP marker is as shown in SEQ ID No.1. The SNP marker detection method comprises the following steps: extracting genome DNA of a miniature pig, which is used as a template, using a primer, the nucleotide sequences of which are as shown in SEQ ID No.2 and SEQ ID No.3, for carrying out PCR amplification, sequencing a PCR amplification product, analyzing a sequencing result, and identifying the G / A polymorphism at the part 325bp of the exon No.1. The SNP marker provided by the invention is susceptibly related to the miniature pig type 2 diabetes, and therefore, miniature pig strains susceptible to or resisting the type 2 diabetes are screened by identifying the SNP marker.

The invention relates to application of scutellarin in preparation of a medicine for treating burns and scalds, and belongs to the technical field of biological medicines. The invention discloses scutellarin (5g / kg) which reduces wound inflammatory response by inhibiting generation and release of inflammatory mediators so as to reduce growth of bacteria on a burn (scald) wound. Meanwhile, the protein and gene expression levels of VEGF-A and TGF-beta1 are up-regulated, so that the expression peak is advanced, the decrustation and hair growing time of miniature pigs is shortened, and the healing rate of wound surfaces is improved. Scutellarin (5g / kg) has an obvious treatment effect on burns (scalds), even has a curative effect better than that of a clinical first-line medicine 1% sulfadiazine silver (10g / kg), and can be used for preparing medicines for preventing and / or treating burns (scalds).

The invention relates to an underwater breathing mask for Bama miniature pigs. The underwater breathing mask comprises a silica gel sleeve (1), a hard shell (2), a first circular supporting plate (3),a second circular supporting plate (7), a plurality of ventilation copper pipes (4) and a plurality of silica gel pads (6); the silica gel sleeve (1) is of a straight cylindrical structure with an opening in one end, and the opening is in a horn shape; the silica gel sleeve (1) is divided into a first rubber sleeve section (8) and a second rubber sleeve section (9); the first rubber sleeve section (8) is nested in the hard shell (2), and a first circular supporting plate (3) and a second circular supporting plate (7) are correspondingly arranged on the outer side and the inner side of the first rubber sleeve section (8) respectively; wherein the hard shell (2) is of a straight cylindrical structure with an opening at one end; and the two ventilation copper pipes (4) are respectively and correspondingly inserted inwards from the round holes on the hard shell (2), and the silica gel pads (6) are arranged between the two ventilation copper pipes (4).

The invention discloses a method for establishing a miniature pig rib fracture fixation animal model and a method for evaluating an absorbable rib fracture fixator by utilizing the animal model. The method for establishing the miniature pig rib fracture fixation animal model comprises the following steps: weighing and performing anesthetic treatment on a miniature pig; lying the miniature pig which is subjected to the anesthetic treatment on the left side to expose a right side, and performing skin preparation treatment at a thorax position; notching for 9 to 11 cm along the parallel position of the sixth rib, cutting intercostal muscles, and bluntly separating a pleura from the rib; biting off an intermediate position of the sixth rib by using a bone ribbing rongeur to obtain the miniature pig rib fracture fixation animal model. By adopting the method for establishing the miniature pig rib fracture fixation animal model, the absorbable rib fracture fixator can be tested and evaluated by the obtained animal model, so that the fracture fixing performance of the absorbable rib fracture fixator is evaluated, and guidance is provided for the research and development of the absorbable rib fracture fixator.