178 results about "Mouse Lung" patented technology

A method for separating and purifying mouse lung microvascular endothelial cell is characterized by comprising using an enzyme to perform digestive treatment on subpleural lung marginal tissue to obtain a single-cell suspension, and performing CD31 immunomagnetic bead sorting to obtain the target cell. Concretely, the method comprises getting subpleural lung marginal tissue of a large suckling mouse, firstly using a mixed solution of collagenase II and neutral protease for digestion to prepare the single-cell suspension, then performing incubation combination with CD31 immunomagnetic bead, then utilizing a magnetic bead sorting purification system to screen CD31+ cell, and removing magnetic beads combined with CD31+ cell by using a magnetic-bead release solution, so as to obtain high-purity mouse lung microvascular endothelial cell. The advantages comprise that the high-purity mouse lung microvascular endothelial cell can be obtained from mouse lung tissue through separation extraction by using enzyme digestion to prepare the single-cell suspension and performing CD31 immunomagnetic bead sorting.

The invention discloses a highly metastatic model of human melanoma, a highly metastatic cell subline of the human melanoma, creation methods for the highly metastatic model and the highly metastatic cell subline, and the dynamic detection of metastasis. The subcutaneously-transplanted mouse highly metastatic model and the corresponding cell subline are established in an in-vivo screening way in a mouse with severe combined immune deficiency (SCID) by using mouse lung metastasis, namely human malignant melanoma cell strain A375 pulmonary metastasis, wherein the highly metastatic cell subline of the human melanoma is A375sci, and has a human tumor cell karyotype; and 60 to 75 hypo-triploid-dominated chromosomes are acrocentric and have a heteroploid karyotype. The cell subline has the two routes of metastasis of blood trails and lymph. The in-vivo screening of the highly-metastatic model is performed by using animals with severe immune deficiency, and is expressed and applied in nude mice. A method for detecting Alu genes by using a polymerase chain reaction (PCR) method is simple, highly sensitive and highly specific, and can be used for detecting organ metastasis, particularly micrometastasis, in a human tumor animal-xenotransplantation model.

The invention relates to a construction method of an immortalized telocytes system. The construction method comprises the following steps: (1) digesting shorn off mouse lung tissue block in type-II collagenase, carrying out filtering and centrifuging, collecting cyte precipitate, culturing the cyte precipitate in a culture medium, after fibrocytes are attached to a wall, transferring supernate to another culture dish, culturing for 12 hours, then changing liquid, and continuing culturing for 3-5 days; (2) taking a trace spearhead as a cyte scraper to scrape off the fibrocytes, and after scraping off the fibrocytes repeatedly, observing a specificity structure telopode and identifying telocytes by an immune marker; and (3) taking the telocytes as host cells, infecting the host cells by using a retroviral vector which contains SV-40-large tumor antigen, and carrying out passage, screening and identifying. Even if the immortalized telocytes system is cultured to the 50th generation, the state is still stable, and therefore, the problem that the purity and the stability of the telocytes cannot be guaranteed by repeated separation and extraction in the prior art.

The invention relates to a preparation method of APP (Actinobacillus Pleuropneumoniae) OMVs (Outer Membrane Vesicles) and a vaccine of the APP OMVs, and belongs to the technical field of biology. The preparation method comprises the steps of culturing APP shope strains in vitro by using a culture medium which is in iron ion limit, obtaining acellular cultural supernatant after carrying out centrifugation and 0.22-[mu]m filtration treatment, and preparing the OMVs released by germs through ultracentrifugation, wherein the observation through a transmission electron microscope shows that the diameter of most OMVs is 50 to 100 nm, the OMVs are used as subunit vaccines to carry out secondary intranasal immunization on a mouse, the weight of the OMV immune mouse is increased for a long time, and the visual forms of lungs have no obvious difference from a PBS (Phosphate Buffer Saline) immune group. Meanwhile, an experiment shows that the OMVs are efficient immune stimulants, not only can high-level IgG (Immunoglobulin G) be stimulated to be generated by mouse sera, but also high-level IgA (Immunoglobulin A) can be generated in mouse lungs, mucosal immunity of the mouse lungs is effectively stimulated, and a better application prospect of using the OMVs as the subunit vaccines is expressed.

The invention provides a method of separating and purifying II-type innate lymphoid cells from animal lung tissue. The method includes: taking the animal lung tissue, crushing, and adding I-type collagenase for digestion to obtain a lung tissue digestion solution; filtering the lung tissue digestion solution; separating to obtain lung tissue lymphoid cells through a Percoll method; using a II-type innate lymphoid cell specific antibody to mark the lung tissue lymphoid cells, and separating to obtain the II-type innate lymphoid cells through flow cytometry. By the method, high-activity II-type innate lymphoid cells can be separated and purified from mouse lung tissue.

The invention discloses a detection method of anti-tumor activity of dendrobium huoshanense refined polysaccharide. The detection method comprises an in-vitro anti-tumor detection method and an in-vivo anti-tumor activity detection method; the inhibition rate of the dendrobium huoshanense refined polysaccharide on tumor cells is calculated through the in-vitro anti-tumor detection method; a dose-effect curve is drawn according to different concentration inhibition rates, and the half inhibitory concentration IC50 of the tumor cells is calculated; the in-vivo growth inhibition effect of the dendrobium huoshanense refined polysaccharide on a C57BL/6J mouse in-vivo transplantable mouse lung cancer LLC tumor is detected through the in-vivo anti-tumor activity detection method; an in-vitro experiment shows that the dendrobium huoshanense refined polysaccharide has an obvious dose effect on an in-vitro anti-tumor effect; an in-vivo activity experiment shows that the dendrobium huoshanense refined polysaccharide has good anti-tumor activity and a relatively good dose-effect relation; the growth of a mouse lung cancer transplantable tumor can be remarkably inhibited and the dendrobium huoshanense refined polysaccharide has an obvious in-vivo anti-tumor effect; two experiment methods are combined so that the detection of the anti-tumor activity of the dendrobium huoshanense refined polysaccharide is more accurate.

The invention relates to application of obakunone to preparation of a medicine for preventing and treating lung injury and pulmonary fibrosis. The medicine consists of obakunone and pharmaceutically acceptable auxiliaries and additives, wherein the obakunone accounts for 3-25% by mass of the medicine. The obakunone in the medicine can inhibit mouse lung injury and pulmonary fibrosis induced by Bleomycin and regulate lymphokine expression related to inflammation of mouse lung injury induced by Bleomycin, and not only is remarkable in effects of preventing and treating lung injury and pulmonary fibrosis, but also has the advantages of low toxicity and no liver or kidney injury.

The invention relates to the fields of tumor models and establishment and application of tumor models and particularly relates to an establishment method and application a radiotherapy model for mousein-situ bearing cancer. The radiotherapy model can be used for evaluating radiotherapy effects. The establishment method comprises the steps of introducing firefly luciferase genes into a mouse Louislung cancer cell strain (LLC) by virtue of a gene recombination technique by taking a wild mouse as a carrier, carrying out subcloning screening so as to obtain a tumor cloning cell strain capable ofstably expressing luciferase, and inoculating recombinant LLC cells to a lung of the mouse. Compared with a traditional tumor model method, the establishment method has the advantages that the pathological and physiological environments of the lung cancer can be stimulated; the method has very important significances to the diagnosis and treatment of the lung cancer; and the model contains luciferase genes and can be used for imaging in a small animal living body imaging instrument, the development process of the lung cancer can be dynamically monitored in real time, the observation is relatively visible and convenient, living bodies can be observed without injuring the animals, the result is relatively reliable, and the model has very good application prospects.

The invention provides application of formononetin in preparing drug for treating pulmonary fibrosis, which is the novel application of formononetin. According to the invention, the formononetin has good effect for pulmonary fibrosis, has no adverse reaction, can slow down the pulmonary fibrosis of mice induced by the bleomycin and has a good application prospect for treating, slowing down or improving the pulmonary fibrosis.

The invention discloses a culture method of mouse lung organs and a special culture solution thereof. The method comprises the following steps: acquiring fresh mouse lung tissue cells, digesting into single cells by collagenase, culturing mouse lung tissue organoid under an in-vitro three-dimensional culture condition, and performing immunofluorescent staining on the lung organoid. The culture solution is prepared from ADF < + + + + >, an RPSO1 conditional culture medium, a Noggin conditional culture medium, a B27 additive, N-acetyl-L-cysteine, nicotinamide, Y-27632, A83-01, SB202190, a recombinant human fibroblast growth factor-7 and a recombinant human fibroblast growth factor-10. Experiments show that the lung organ established by the method disclosed by the invention can effectively maintain tissue cell specificity, stem cell characteristics and biological functions, and meets the requirements of scientific research.

The invention relates to application of demethylwedelolactone-7-sulfate in preparation of an anti-pulmonary fibrosis drug. The compound demethylwedelolactone-7-sulfate is obtained from traditional Chinese medicine herba ecliptae for the first time and the structure of demethylwedelolactone-7-sulfate is determined through NMR (Nuclear Magnetic Resonance) and other wave spectrum data, the in-vivo and in-vitro pharmacological experiments prove that the demethylwedelolactone-7-sulfate can obviously inhibit the proliferation of TGF-beta1 induced human embryo lung fibroblasts and the HYP (hydroxyproline) content, the inflammation and fibrosis degree of lung tissue in bleomycin induced mice pulmonary fibrosis model can be reduced after the anti-pulmonary fibrosis drug is orally taken at a dosage of the 5-30mg/kg, the content of MDA (methylene dioxyamphetamine) reflecting the lung injury degree in lung tissues is influenced, the content of HYP reflecting the collagen deposition of lung tissue can be reduced, the content of cell factor TGF-beta1 causing pulmonary fibrosis can be reduced, and the like, so that the pharmacological action for improving the pulmonary fibrosis degree can be achieved. The demethylwedelolactone-7-sulfate has new applications as a drug for treating or improving pulmonary fibrosis disease.

The invention discloses a method for establishing a mouse lung cytomegalovirus latent reactivation infection model. The method comprises the following steps: injecting MCMV into the abdominal cavity of a mouse to establish mouse lung latent infection cytomegalovirus, selecting latent MCMV-infected mouse, orally taking a medicine prednisone to induce reactivation of the lung latent cytomegalovirus,and identifying reactivation of the lung latent cytomegalovirus. Lung latent cytomegalovirus infection is induced to be reactivated by orally taking the medicine prednisone, the mouse lung is subjected to cytomegalovirus latent reactivation infection, and an animal model with cytomegalovirus latent reactivation can be constructed. The mouse model shows the typical characteristic of lung cytomegalovirus latent reactivation that: a virus expression-related protein appears in lung tissues. The model can be used for fundamental research in the related fields of cytomegalovirus infection, and laysan experimental animal foundation for exploring the pathogenesis of latent reactivation of cytomegalovirus.

The invention provides an economic, simple and convenient lung organ culture method and an experimental model, and relates to the technical field of biology. The method comprises the following steps: marking a transgenic mouse with pulmonary epithelial cells by using GFP (Green Fluorescent Protein), sorting the pulmonary epithelial cells and interstitial cells by using a flow cytometry, then co-culturing the sorted lung epithelial cells and mesenchymal cells in an upper-layer small chamber and a lower-layer small chamber divided by Transwell, and promoting the epithelial cells in the upper-layer small chamber to grow and develop into lung organs by using growth factors and biological components secreted by the mesenchymal cells in the lower-layer small chamber. According to the method of the invention, the natural growth environment and mode of in-vivo cells are simulated, and the lung bronchial organ can be obtained under the condition that exogenous growth factors are not added. The mouse lung organ can be used as the experimental model, and can be used for research on lung development process, cell differentiation, lung toxicology, drug screening, organ regeneration and transplantation, etc.

The embodiment of the invention provides an application of zeartinib in preparation of drugs for treating pulmonary fibrosis diseases, and the structural formula of zeartinib is shown in the specification. Experiments show that zeartinib has a good effect on pulmonary fibrosis, has no adverse reaction, can slow down bleomycin-induced pulmonary fibrosis in mice, and provides a good application prospect for the treatment, alleviation or improvement of pulmonary fibrosis diseases.

The invention belongs to the technical field of medicines, and particularly relates to new application of guanosine, in particular to new application of guanosine in preparation of medicines for treating asthma or inhibitors of MAPK, NF-kappa B and STAT3. The invention aims to solve the problem that safer and more effective asthma treatment medicines need to be developed urgently in the prior art.It is found that in an in-vitro THP-1-derived macrophage inflammation model, guanosine inhibits generation of a proinflammatory factor IL-6 by inhibiting activation of MAPK and NF-kappa B; in an asthma mouse model, guanosine reduces OVA-IgE of mouse plasma, reduces generation of IL-4, IL-6 and IL-13, relieves airway high reactivity, and relieves lung tissue cell infiltration, airway inflammationand collagen deposition. In addition, the expression levels of p-p38 MAPK, p-p65 NF-kappa B, p-I kappa B alpha and p-STAT3 proteins in lung tissues of mice in a guanosine treatment group are obviouslylower than those in an asthma model group. Therefore, the invention provides application of guanosine in preparation of a p38 MAPK inhibitor, a p65 NF-kappa B inhibitor, an STAT3 inhibitor or an asthma treatment drug.

The invention relates to a mouse lung tissue primary epithelial stem cell ball culture method. The method comprises the following steps: digesting a mouse lung lobe tissue block by adopting a preheated collagenase digestion solution, adding an equal amount of FBS-containing DMEM/ F12 culture medium to terminate digestion, extracting mouse lung tissue primary epithelial stem cells, and carrying outsuspension culture to form stem cell balls. The mouse lung tissue primary epithelial stem cell ball culture method is established through a 3D culture model, so that a lung tissue microenvironment issimulated in vitro, and an effective and reliable research model is provided for clarifying a regulation mechanism of lung epithelial stem cell proliferation and differentiation.

The invention belongs to the technical field of Chinese medicine lung cancer syndrome and relates to a building method for a lung-kidney deficiency type lung cancer mouse model. The lung-kidney deficiency type lung cancer mouse model can be used for preparing an animal model of lung-kidney deficiency type lung cancer disease combination. In the model, a preparation method that a mouse lung Qi deficiency and kidney deficiency model and a Lewis lung cancer cell transplantation tumor model are used for joint molding is adopted, i.e., a Lewis lung cancer cell is subjected to subcutaneous injection to build a lung cancer transplantation tumor model, a smoked stimulation method is adopted for building a lung Qi deficiency model, and hydrocortisone is subjected to intraperitoneal injection to build a lung deficiency model. The model is simple to operate, has the advantages of short molding period, high success rate and good repeatability and stability, and withstands the measurement and verification by a prescription (medicine). Most importantly, the model conforms to the pathologic features of lung cancer and has the characteristics of the Chinese medicine theory. The Chinese medicine theoretical knowledge can be combined to comprehensively analyze the scientificity of the mouse lung-kidney deficiency type lung cancer animal model, thus a scientific research tool is provided for researching the lung cancer resistance of the traditional Chinese medicine.

The invention provides a method for efficiently inducing advanced branched pulmonary organs in vitro, an experimental model and a compound combination, and relates to the technical field of biology. The method is to digest airway epithelial tissues in vitro, then carrying out multiplication culture on the airway epithelial cells by utilizing an amplification culture medium containing epidermal growth factors and insulin, digesting the airway epithelial cells into single cells by utilizing pancreatin, and then adding the single cells into a differential culture medium for culture to obtain a mouse lung organ which has a branched advanced structure and contains differentiated ciliated cells. The mouse lung organ can be used as an experimental model for precision medical treatment, organ transplantation, drug screening and mechanism research on drug action. According to the method disclosed by the invention, the airway epithelial cells can be cultured into the lung organs which have branched advanced structures and differentiated ciliated cells that are more similar to the lung morphology and cell structure in vivo, and the yield of the cultured lung organs is more than three times that of the lung organs cultured by a conventional method.

The invention discloses application of ginsenoside Rg3 in combination with GP as an antitumor drug. A mouse Lewis lung cancer cell line (LLC) is selected and inoculated to the right armpit of a C57BL/6N mouse, and the influence of ginsenoside Rg3 and Rg3 in combination with GP on Lewis lung cancer is observed; hematoxylin-eosin staining is performed on the liver, kidney, spleen and tumors, and pathological changes of ginsenoside Rg3 in combination with GP on the liver, kidney, spleen and tumors of mice with the Lewis lung cancer are detected; and tumor tissues are subjected to immunohistochemical staining, and the density of blood vessels in tumors and the expression of tumor platelet surface activation markers are detected. The conclusion is that the ginsenoside Rg3 in combination with GPcan enhance the effects of ginsenoside Rg3 of inhibiting the tumor growth of the mice with the Lewis lung cancer and inhibiting tumor angiogenesis, and the tumor inhibition effect of the ginsenosideRg3 is possibly and closely related to CD62P expression reduction; and the ginsenoside Rg3 in combination with GP can achieve the synergistic and toxicity-reducing effects.

The invention provides a construction method and application of a visual mouse model for monitoring functions of specific CTLs of SARS-CoV-2 Spike protein S1, and belongs to the technical field of animal model construction. According to the invention, firstly, a recombinant expression vector containing an eukaryotic promoter, a Spike protein S1 gene and a fluorescence reporter gene is constructed, the recombinant expression vector is transfected into a mouse lung through a transfection reagent, transcription and translation levels of the S1 gene and the fluorescence reporter gene in a mouse body are consistent, after a mouse is infected with SARS-CoV-2, immune response is initiated in the body, cytotoxic T cells that specifically kill the S1 protein are produced to clear the infected lung cells, and the fluorescence report protein is also cleared along with the S1 protein, so that an in-vivo fluorescence intensity of the mouse can be monitored in real time by using an in-vivo fluorescence imaging system. Therefore, a function strength of the CTLs in a mouse body can be evaluated by monitoring a fluorescence intensity in real time. The constructed visual mouse model provided by the invention can be used for screening and evaluating drugs or vaccines of SARS-CoV-2.