90 results about "Tree shrew" patented technology

The present invention discloses a method for evaluating a cigarette harm degree by using primate animal models, and belongs to the field of cigarette toxicology evaluation and related technology detection. According to the method, Chinese tree shrews are adopted as test animals, an active smoking device with a special design is adopted to enable tested animals to be at a smoking stimulating state so as to observe conditions of the animals at the state, determine real test data, and reasonably evaluate a cigarette harm degree. The method of the present invention has the following advantages that: the specific Chinese tree shrews with high biological classification are adopted to carry out evaluation researches of tobacco toxicology and attenuated mechanisms, wherein the method has characteristics of sensitive response on toxicological effects produced by cigarette smoke gas, convenient operation and economic breeding, the evaluation system used in the method is perfect, the evaluation cost is relatively low, the evaluation result obtained by using the primate animal models provides comparative medicine values compared with the evaluation result obtained by using other experimental animals, and the method is applicable for evaluation of cigarette harm reduction efficacy.

The invention provides a fluorescent staining method of evaluating sperm motility of a tree shrew. The fluorescent staining method comprises the following steps: (1) preparing a TALP-HEPES sperm rinse; (2) collecting sperms; (3) washing the sperms; (4) staining; and (5) detecting by a microscope. According to the fluorescent staining method provided by the invention, by utilization of the characteristics that Hoeches 333422 and PI are different in reaction to dead and alive sperms and can be excited by a laser with a same wavelength, fluorescent staining is carried out on sperms of the tree shrew, and the sperm motility can be accurately detected without needing to switch a light source; in addition, as the two dyes respectively emit red and blue fluorescence after being excited by the laser, a dye which emits green fluorescence can also be introduced to detect the integrity of sperm acrosomes while the motility is detected, and the sperm motility and the integrity of the sperm acrosomes of the tree shrew are accurately detected by virtue of a fluorescence microscope or a flow cytometer.

The invention relates to a method for detecting and estimating the change situation of CD81 molecule which is infected by hepatitis C virus (HCV) in vivo and in vitro, in particular to a Taqman fluorescence probe quantitative PCR detection method on the basis of using cloned tree shrew CD81 plasmid as standard substance, belongs to the molecular biology field. The method comprises the following steps: cloning tree shrew CD81 gene segment; designing reasonable probe and primer with Primers Express2.0 software on the cloned tree shrew CD81 gene segment; designing experiment to select annealing temperature (Tm value), primer concentration and probe concentration under optimal conditions; further adopting tree shrew CD81 plasmid as standard substance to perform 10 times concentration gradient dilution and building standard curve; verifying the repeatability, stability and sensitivity of the method of the invention; and finally detecting the change of CD81 molecule which is infected by hepatitis C virus (HCV) in vitro by the method of the invention. The method can detect the change situation of tree shrew CD81 accurately, fast and conveniently.

The invention relates to a tree shrew hepatocyte immortalized cell line and its construction method and application and belongs to the technical field of biomedicine. The cell line is named as ITH6.1, is preserved in the China center for type culture collection in the Wuhan University at No. 229, Bayi road, Wuchang District, Wuhan City, Hubei Province and has a preservation number CCTCC NO: C201740. After continuous passage 100 times, the tree shrew hepatocyte immortalized cell line ITH6.1 still keeps a diploid karyotype and has a very strong multiplication capacity so that immortalization is realized really. The cell line has a strong multiplication capacity and realizes protein expression and biochemical index characteristic analysis of ITH6.1. The result shows that the ITH6.1 has typical tree shrew hepatocyte characteristics. The safety test such as a tumorigenicity test also indicates that the cell line has better biosecurity.

The invention relates to a construction method for an acute hyperuricemia tree shrew model. The method comprises the following steps: (1) selecting adult healthy tree shrew with body weight of 110 to 150 g and an age of 1 to 3 years; (2) respectively measuring fasting serum uric acid values of males and females of tree shrew selected in the step (1); (3) carrying out conventional breeding on the tree shrew selected in the step (1) and pretreating the tree shrew by grasping and releasing the tree shrew with hand; and (4) injecting the tree shrew treated in the step (3) with an oteracil potassium suspension so as to obtain the acute hyperuricemia tree shrew model, wherein the amount of the used oteracil potassium suspension is 40 mg/kg to 100 mg/kg on the basis of the body weight of the tree shrew. The model has typical disease symptoms, a quantity index has statistical significance, and serum urea nitrogen and serum creatinine values and histopathological results prove that such a model making dosage range is safe and does not result in influence on the liver and the kidney.

The invention discloses an in-vitro separation and purification method for tree-shrew corneal endothelial cells. The in-vitro separation and purification method includes the steps that tree-shrew corneal endothelial cells are separated, purified and subjected to passage cultivation through CECs, and corneal endothelial cells are obtained. The separated corneal endothelial cells have high cell activities, purification of the corneal endothelial cells can be achieved in the passed P3 generation is passed, the purified cells have typical endothelial cell morphology, and are identified through NSE immunofluorescence, and the dyeing rate is high. The purification efficiency of the method is greatly improved, the cost is low, using is convenient, and the large quantity of tree-shrew corneal endothelial cells can be obtained. In-vitro cultivation of the tree-shrew corneal endothelial cells is achieved, and the void that a specific corneal-endothelial-primary-cell in-vitro cultivation technology for a tree-shrew species does not exist ate present is filled; the material taking and purification methods of primary cells are easy to operate, the cost is low, and in-vitro cultivation tree-shrew corneal endothelial cells can be kept the good cell states in the long-term passage cultivation process.

The invention relates to a complete pellet feed for tree shrews and a preparation method thereof. The complete pellet feed comprises the following raw material components in percentage by weight: 38 to 40 percent of corn, 17 percent of barley, 10 percent of rice, 17 percent of bean pulp, 8 percent of fish meal, 2 percent of alfalfa meal, and 1.2 percent of egg powder which are taken as main raw materials, and 1 percent of sucrose, 0.5 percent of fructose, 1.5 percent of beef tallow, 1 to 1.5 percent of lard, 0 to 1.5 percent of whole milk powder, and 0.8 percent of amino acid, vitamin and trace element which are taken as additives. The raw materials are prepared into the pellet feed which has the diameter of 0.5cm and the length of 0.5 to 1.2cm. The pellet feed of the invention has overall and balanced nutrition, so that the stress reaction of the tree shrews caused by feed factor is avoided. The feeding is convenient, the requirements on the normal growing development of the tree shrews under laboratory conditions can be met, and the normal oestrus, mating, impregnation and lactation of the tree shrews all the year round are ensured.

The present invention provides a method for detecting the transcription level of a tree shrews gene SLC22A12/UTAT1 through RT-qPCR. According to the method, the total RNA is extracted from the fresh kidney tissue of tree shrews, conventional reverse transcription synthesis is performed to obtain a tree shrews kidney tissue cDNA first strand as a template, real-time fluorescence quantitative PCR amplification is performed with a PCT primer combination, the Ct values, the dissolving peaks and the amplification efficiencies of the gene SLC22A12/UTAT1 fragment and the internal reference gene GAPDHfragment are obtained, and the [delta][delta]C(t) value expressed by the normalized gene SLC22A12/UTAT1 folds is obtained through treatment so as to obtain the relative expression value of the gene SLC22A12/UTAT1 transcription level. According to the present invention, the method provides the effective way for the research on the function of tree shrews urate anion transporter 1 and the influencecaused by related drugs, is suitable for the real-time quantitative PCR detection, and has advantages of simple operation, high reproducibility, low detection cost, high sensitivity and strong specificity.

The invention discloses a tri-antibody sandwich ELISA (enzyme-linked immunosorbent assay) detection method of the IgG (Immunoglobulin G) of a tree shrew, which is applicable to the detection of the total content of IgG in the blood plasma of the tree shrew. The method comprises the following steps: coating an enzyme-label plate with the monoclonal antibody in the anti-rat IgG of the tree shrew as a capture antibody, and combining with the IgG of the tree shrew to be detected; and establishing the tri-antibody sandwich detection method of the IgG of the tree shrew by optimizing the concentration of the antibody and other multi-factor conditions by using the polyclonal antibody of the anti-rabbit IgG of the tree shrew as the secondary antibody and the commercial goat anti-rabbit IgG-HRP as the detection antibody. In the invention, the IgG level of the blood plasma of 30 tree shrews is detected by using the method, wherein according to the detection, the normal IgG level of the blood plasma of the tree shrews is 8.6g/L-17.8g/L. The tri-antibody sandwich ELISA detection method of the IgG of the tree shrew provides necessary tools for the research on the tree shrew as the animal model for the experiment on human diseases and the immunity mechanism of the relates diseases.

The invention discloses a tree shrew active behavior monitoring box, and relates to the technical field of animal experiment scientific research test and analysis equipment. Detachable wall baffles are inserted into plug slots of a white base plate with a groove, an organic glass transparent top cover is fixedly installed at the upper ends of the detachable wall baffles, the detachable wall baffles comprise the front transparent organic glass front baffle, the non-transparent white left baffle, the non-transparent white right baffle and the non-transparent white rear baffle with the back faceprovided with a reinforcement bar, the left baffle and the right baffle are provided with reinforcement bars, and plug slots are formed in the inner side walls of the non-transparent white left baffleand the non-transparent white right baffle which are provided with the reinforcement bars. Accordingly, the operation processes such as installation, operating and cleaning are easy, the tree shrew activity can be accurately measured, and the high repeatability is achieved; tree shrew capturing is not needed in the whole process, and artificial stress caused by grabbing to a tree shrew is eliminated; while the tree shrew activity is accurately measured, the natural instincts of the tree shrew good at jumping can be recovered to the maximum extent.

The invention provides a method for establishment of an acute lung injury model of tree shrews and application of the model. After the tree shrews are narcotized, 180-200 mg/kg of a lipopolysaccharidesolution is intratracheally administrated, and when the lung injury area of the tree shrews reaches a half or more, the acute lung injury animal model of the tree shrews is established. Through the establishment method, lung injury of the tree shrews can reach a half or more after 70 hours; in 96 to 120 hours, the tree shrews clinically show the symptoms of wasting, breathing difficulties and thelike, the lung injury area of the tree shrews is 70% or more, and the blood oxygen index can reach 200 mmHg or below. The modeling method is efficient and high in success rate, and the cost can be effectively saved; the model cycle is 70 hours or more, and the research cycle space for acute lung injury is significantly widened; and the established tree shrew animal model is incapable of self-healing and reversible recovery, the clinical symptoms are more similar to human diseases, and good prospects of industrialization are achieved.

The invention discloses a method for establishing an EB virus tree shrew animal model. The method for establishing the model comprises the steps of animal selection, preparation of a virus liquid, innoculation, observation of the state of a tree shrew, periodical blood drawing after innoculation for the tree shrew, separation of PBMCs, measuring of EBV copy numbers in PBMCs through qPCR, measuringof expression of relevant EBV genes in PBMCs through RT-PCR, and pathological examination. The method for establishing the EB virus tree shrew animal model is disclosed for the first time, and a basis is laid for better researching EB virus biological characteristics and diseases relevant to the EB virus; the selected tree shrew is a novel experimental animal, and is closer to primates than currently-existing experimental animals; moreover, the tree shrew has the advantages of being susceptible to various viruses, easy to feed, small in size, easy to obtain and the like.

The invention provides a method for detecting the transcription level of tree shrew xanthine dehydrogenase/oxidase gene through RT-PCR. The method comprises the following steps: carrying out PCR amplification by using a PCR primer combination with cDNA obtained after inverse transcription of total RNA of a sample as a template to obtain a ratio of the gray value of an amplification product of an XDH/XO gene fragment to the gray value of a PCR amplification product of an internal control gene GAPDH, calculating the relative expression value of XDH/XO mRNA under different physiological conditions according to the gray value ratio, and semi-quantitatively detecting the transcription level of tree shrew xanthine dehydrogenase/oxidase gene according to the calculated relative expression value in order to provide an effective way to study the functions of tree shrew xanthine oxidase and the influences of related drugs on the tree shrew xanthine oxidase. The method is suitable for the semi-quantitative RT-PCR detection, and has the advantages of low cost, simple operation, high repeatability low detection cost, and good specificity and sensitivity.

The invention discloses a tree shrew behavior sensitization model evaluation method and an open field experiment device for the evaluation, the method comprises the following steps of: inducing the tree shrew behavior sensitization by METH, detecting the tree shrew behavior science, the motion distance, the speed and the track by an open field experiment, measuring the weight change of the tree shrew and marking plate behaviors; the invention also discloses an open field experiment device for evaluating the tree shrew behavior sensitization model, which comprises a box body, a bottom plate replacement box, a moving chassis, a fixing bracket and a camera, wherein the bottom of the moving chassis is provided with universal wheel with a brake, the box body is fixed on the top of the bottom plate replacement box, the bottom plate replacement box is fixed on the moving chassis, an upright post of the fixing bracket is fixed on the moving chassis, and the camera is fixed on the fixing bracket. The device provides reference and guidance for the success of establishing the tree shrew behavior sensitization model, and is convenient for researching the addiction mechanism of lazy drugs by means of the tree shrew behavior sensitization model; the open field experiment device can conveniently replace the bottom plate after each batch of experiments are finished, thereby reducing the time for cleaning, scraping, assembling and other processes.

The invention relates to a method for establishing an HCV (hepatitis C virus) cell model by utilizing tree shrew bone marrow mesenchymal stem cells, and belongs to the technical field of biological medicines. The method comprises the steps that firstly, an OCLN (occludin) lentiviral vector is adopted to infect the tree shrew bone marrow mesenchymal stem cells, then a CD81 lentiviral vector is added for further infection, then a miR-122 lentiviral vector is added for infection for 4 hours, then a fresh culture medium containing 4 mcg/mL of polybrene with 1/2 volume of a current culture medium is added, and the used culture medium is removed the next day and is replaced with the fresh culture medium for continuous culture so as to obtain the HCV (hepatitis C virus) cell model. According to the method, the cell model has the basic characteristics and the differentiation potential of the bone marrow mesenchymal stem cells and is susceptible to HCV (hepatitis C virus), HCV (hepatitis C virus) endocytosis and copy are supported, infective virus particles are produced, and new cell model resources are provided for HCV (hepatitis c virus) pathogenesis researching and drug screening.

The invention discloses a tree shrew behavioristic evaluation method. The method combines the method for hiding the food of a tree shrew by using a food hiding box, an experimental tree shrew selecting and training method and a study and memory scoring method. The characteristic that the tree shrew is developed in vision is utilized, the food of the tree shrew is hidden in the food hiding box, color marking and training are carried out, a whole experiment is carried out under the conditions that no stimulation influence factor exists, and comparison on behavior changes of the tree shrew is carried out through the average score of the tree shrew before and after the experiment is started. More scientific, comprehensive and objective evaluation results can be obtained through the tree shrew behavioristic evaluation method. The used materials are cheap and easy to manufacture, operation is convenient, and the evaluation method is scientific.

The invention discloses a tree shrew oral cavity expander for experiments, and mainly relates to the technical field of tree shrew experimental equipment. The tree shrew oral cavity expander comprisesa body sleeve, an elastic band and an adjustable opening supporting device, the adjustable opening supporting device comprises a supporting plate and an adjuster; an observation opening is formed inthe center of the supporting plate; the supporting plate comprises an upper supporting plate and a lower supporting plate, and the upper supporting plate and the lower supporting plate are rotationally connected through a rotating shaft; the adjuster comprises a fixed rod cylinder hinged to the tail end of the lower supporting plate and an adjusting rod cylinder hinged to the tail end of the uppersupporting plate. The tree shrew oral cavity expander has the beneficial effects that the tree shrew oral cavity expander is invented based on a new opening supporting device technology, has a betterfixing effect, is easy, convenient and efficient to use, and can adjust the supporting height according to the oral cavity size and experiment requirements; moreover, the mouth opening amplitude of animals can be slightly adjusted, and the animals can slightly adjust the mouth opening amplitude to improve the comfort level. Furthermore, the tree shrew oral cavity expander is not only suitable fortree shrews, but also suitable for all small experimental animals.

The invention relates to a tree shrew retina ischemic injury experiment model adopting photochemical induction, and belongs to the technical field of clinical medicine, pharmacy and cell biology. The experiment model is obtained through steps as follows: a healthy adult tree shrew is selected and is anaesthetized by 40 mg*kg(-1) of 2.5% sodium thiopental through intraperitoneal injection; the tree shrew is fixed on an operating table, and 2.75% rose Bengal normal saline is injected from the sublingual vein of the animal in a dose of 0.031 ml/10 g; 2 drips or 3 drips of a 0.25% compound tropicamide solution are utilized to dilate pupils; after rose Bengal is circulated for 10 min, a small animal cerebral thrombosis experimental device with the patent number of ZL201420068737.2 is adopted to irradiate a left eyeball or a right eyeball, so that vascular endothelial injury and platelet aggregation induce thrombogenesis, and the surface temperature of the irradiated part is controlled to be 37.0 DEG C plus or minus 0.1 DEG C; the thermal insulation is performed, the animal is placed back to a cage after coming round, fundus retina photographing is performed after the experiment is finished for 4 h, 24 h and 72 h; fundus artery thrombogenesis and retina injury change are dynamically observed with a conventional method. The model has the advantages of simplicity and convenience in operation, good repeatability and low experiment cost.

The invention discloses a method for constructing, observing and analyzing a tree shrew fusarium type keratitis model. The method comprises the following steps: fusarium culture, fusarium spore preparation, experiment grouping and fusarium type keratitis modeling, fusarium type keratitis model culture analysis and fusarium type keratitis observation. According to the invention, the eyes of the tree shrew are used as experimental eyes, and the construction and analysis observation of the fusarium type keratitis model are carried out, so that the construction and timely and effective observationand analysis of the fusarium type keratitis model can be realized, and the popularization and application are facilitated.

The invention relates to a non-animal origin tree shrew sperm in vitro capacitation liquid and application thereof, belonging to the technical field of animal reproductive biology. The specific stepsare as follows: preparing tree shrew ishing semen and capacitation liquid; separating and removing the epididymis cauda of tree shrews; placing the epididymis cauda in centrifugal tubes containing ished semen and incubating for 10 min to allow sperm to swim out. transferring the sperm suspension to the sterilized centrifuge tube, adding the semen ishing liquid, gently mixing, and centrifuging; discarding supernatant, adding semen ishing liquid, and centrifuging; discardingthe supernatant and resuspending the sperm in the capacitation solution to make the sperm concentration 10* 106 sperm/mL.5 ml of that semen is divided into incubation tubes and incubating in a CO2 incubator. A method for in vitro capacitation of tree shrew spermatozoa adopting animal-free component capacitation liquid avoids that risk of introduce pathogenic factors into in vitro capacitation or in vitro fertilization system, and is easy to carry out quality control on the capacitation liquid.