36 results about "Murine cell" patented technology

The present invention relates to an RNA capping enzyme comprising or consisting of an amino acid sequence having high identity with any of the sequences of SEQ ID NO: 1-28, which can be expressed in vitro, in human cells, in mouse cells or in yeast cells, and which has the activities of RNA 5 '-triphosphatase (RT), guanylate transferase (GT) and RNA guine-N7 methyltransferase (RNMT) at the same time. The invention also relates to a mutant obtained by carrying out substitution, addition and / or deletion of amino acid on any sequence in SEQ ID NO: 1-28, a nucleotide sequence for coding the capping enzyme and / or the mutant thereof, and an expression construct or a host cell containing the nucleotide sequence. The invention also relates to a composition comprising the capping enzyme as well as a method and a reaction system for capping RNA by using the capping enzyme, a mutant or a composition thereof.

The invention relates to the technical field of medicines and biology, in particular to a mouse EGFR (Epidermal Growth Factor Receptor) mutant lung adenocarcinoma cell line (EP Mus musculus) as well as preparation and application thereof. The invention provides an EP cell line ([EGFRE19delta (mutation); trp53 (deletion), EP] is derived from a lung adenocarcinoma model of a C57BL / 6 strain mouse, carries the most common EGFRE19del mutation in lung adenocarcinoma, is a first lung adenocarcinoma mouse cell line carrying EGFR mutation known in the world at present, has the in-vivo tumorigenesis ability of the C57BL / 6 strain mouse, and fills the blank of the mouse EGFR mutant cell line. The cell line provided by the invention has the background of EGFRE19del homozygous activation mutation and Trp53 homozygous deletion mutation, is sensitive to EGFR-TKI targeted therapy, can be used for in-situ tumor formation in subcutaneous and lung of an immune normal mouse C57BL / 6, and is suitable for immunotherapy and targeted therapy research of EGFR mutation lung adenocarcinoma.

The utility model relates to the field of culture devices, in particular to a partitioned culture device for mouse cells. Comprising a culture dish body, a plurality of culture cavities are formed in the upper surface of the culture dish body, an adjusting structure is arranged on one side of the culture dish body and comprises a sealing film and a connecting plate, the inner wall of the sealing film is attached to the upper surface of the culture dish body, and two rolling wheels are rotationally connected to one side of the connecting plate; and the arc surface of the rolling wheel is in transmission connection with two belts. According to the partitioned culture device for the mouse cells, provided by the utility model, a person can conveniently perform mouse cell culture operation in a single culture cavity, so that all the culture cavities are prevented from being exposed outside, and the environment and temperature in the culture cavities can be prevented from being damaged; only one culture cavity is exposed, so that the risk that pollutants in the air enter other cavities can be reduced, and the success rate of mouse cell culture is improved.

The invention belongs to the technical field of biological medicine, and relates to pharmaceutical application of an oxidative phosphorylation inhibitor, in particular to application of the oxidative phosphorylation inhibitor in preparation of an immunomodulatory drug, and the immunomodulatory drug has the advantages that the function damage of newborn myeloid-derived suppressor cells caused by antagonism BCG (bacillus calmette guerin) vaccination is inhibited, and the immunomodulatory effect is improved. The compound is used for preventing or treating neonatal immune-related diseases caused by bacillus calmette guerin vaccine inoculation. The invention finds that the BCG can cause damage to the immunosuppression function of the MDSC of the suckling mouse through up-regulation oxidative phosphorylation, and the negative effect caused by inoculation of the BCG can be reversed by using the OXPHOS inhibitor, so that the immunosuppression function of the MDSC of the suckling mouse is successfully recovered in an in-vivo and in-vitro model, and the immune-related adverse reaction caused by inoculation of the BCG is expected to be improved.

The invention provides a mouse PDX cell and a construction method of a humanized systemic lupus erythematosus (SLE) mouse model, and belongs to the technical field of animal model construction. The invention provides a mouse PDX cell L0407-6-P3 which is preserved in the China Center for Type Culture Collection (CCTCC), and the preservation number is CCTCC NO: C2024181. After the mouse PDX cell L0407-6-P3 is intravenously injected into an NSG mouse, the mouse disease is stable, the human SLE disease phenotype can be generated, the multi-system and multi-organ inflammatory response and injury of human SLE can be better simulated, the mouse PDX cell L0407-6-P3 can be used for constructing a humanized systemic lupus erythematosus mouse model, the blank of the current humanized SLE animal model is filled, and the mouse PDX cell L0407-6-P3 has better popularization and application values.

The invention discloses application of 5 beta-cholanic acid (5 beta-CA) in enhancement of NK cell functions and preparation of antiviral drugs. The NK cell function of a chronic hepatitis B (CHB) patient is depleted, it is found that the intracellular 5beta-CA level is reduced by screening differential metabolites of peripheral blood NK cells of the CHB patient, and the NK cell function of the CHB patient can be enhanced by exogenous supplementation of 5beta-CA; the function of NK cells of HBV persistently infected mice can be enhanced, and the level of virology indexes such as serum HBV related antigens and the like can be reduced. Specifically, the 5beta-CA enhances the NK cell function by activating a TGR5-cAMP-mTOR signal channel so as to play an anti-HBV role. The research finds that the improvement of the 5beta-CA level is expected to become a new thought for enhancing the NK cell function of a CHB patient, and a new technical means is provided for the treatment of CHB.

The invention provides the primer and the detection kit for detecting the culicoides chromogen immune mouse B cell receptor repertoire, the amplified B cells and plasma cell clone can be identified and quantified by screening the BCR repertoire by utilizing the kit, and the kit has the advantages of high throughput, rapid and accurate detection and the like. Experiments show that a basis is provided for the research on the immune mechanism of the culicoides leucoides allergen.

The invention discloses application of sulforaphane, and belongs to the field of medicine. Experiments with mouse AML-12 cells find that when the mouse AML-12 cells are jointly treated with sulforaphane and alpha-amatoxin, the inhibition effect of alpha-amatoxin on the activity of the AML-12 cells can be effectively relieved, the ROS level in the cells can be reduced, cell oxidative stress injury can be relieved, mitochondrial membrane potential reduction can be inhibited, and the mouse AML-12 cells can be treated with raphane and alpha-amatoxin. The sulforaphane can be used for relieving alpha-amatoxin induced mouse AML-12 cell mitochondrial injury, so that the sulforaphane can become a novel medicine for relieving alpha-amatoxin poisoning. The new application of the sulforaphane is found for the first time, and a new substance is provided for preparing the medicine for relieving alpha-amatoxin poisoning.

The new alpharetrovirus-based particles are suitable for high efficiency of transiently transducing animal cells. e.g. human or murine cells, and which efficiently introduce coding and non-coding RNA contained in the alpharetrovirus-based particles into target cells The alpharetrovirus-based particles also provide for high efficiency of the activity and / or integrity of the RNA that is introduced into the animal cells, as the particles protect the incoming RNA from degradation during entry. The transferred RNA can be of non-coding nature (e.g. single guide (sg) RNA. short-hairpin (sh) RNA. micro RNA. or long non-coding (Inc) RNA. or the RNA may encode proteins or peptides. e.g. receptors. transcription factors. cellular enzymes, antigens for use in vaccination, gene / protein therapy and / or gene editing nucleases, recombinases and transposases.

The invention provides FAP CAR mRNA, a polymer lipid nanoparticle delivery system and a preparation method and application thereof, and belongs to the technical field of biological medicine. The FAP CAR mRNA polymer lipid nanoparticle delivery system disclosed by the invention comprises FAP CAR mRNA, an RP182 polypeptide and a lipid nanoparticle. The FAP CAR mRNA polymer lipid nanoparticle delivery system which is uniform in particle size and good in stability is successfully prepared, the mRNA encapsulation efficiency is high, mouse RAW264.7 cells can be efficiently transfected, FAP CAR is expressed by macrophages, and the transfection efficiency is remarkably higher than that of the FAP CAR mRNA or other control vectors independently used. The macrophages (CAR-M cells) transfected by the FAP CAR mRNA delivered by the LNP can specifically recognize and kill the activated fibroblasts, and the proliferation of the fibroblasts is remarkably inhibited.

To provide agents and their use in the treatment of endocrine, nutritional and / or metabolic diseases in a mammal.SOLUTION: The present disclosure provides novel peptides which stimulate β-cell proliferation, have ability to rescue β-cells from apoptosis induced by glucotoxic conditions, and stimulate insulin secretion from rat INS-1 β-cells as well as isolated mouse pancreatic islets.SELECTED DRAWING: Figure 1

The invention discloses an application of a Pik3cb inhibitor in treatment of Sjogren's disease. Experiments prove that the Pik3cb inhibitor TGX-221 can reduce the number, activation and chemotaxis of NOD mouse B cells of a Sjogren's disease model mouse, plasma cell differentiation and antibody secretion, improve the immune microenvironment of the submaxillary gland of the Sjogren's disease model mouse, reduce the infiltration degree of submaxillary gland lymphocytes and promote the salivary secretion function; and after the Pik3cb is over-expressed, the symptom and pathology of the Sjogren's disease are aggravated. Therefore, the Pik3cb inhibitor can be used for treating the Sjogren's disease. The invention provides a new thought for research and development of medicines for treating the Sjogren's disease, and has important application value.

The invention relates to the technical field of immune cells, and discloses a rat NK cell separation and multiplication culture method which comprises the following steps: screening rat PBMC cells in a CD161 + positive screening mode to obtain rat NK cells; the rat NK cells and the rat PBMC cells are mixed and co-cultured, and the working concentration of the interleukin growth factor is adjusted in stages in the culture process. In the scheme of the application, the rat NK cells obtained by separation and amplification in the peripheral blood of the rat have the characteristics of large quantity, strong functional activity, no infectious pathogen pollution and the like, and can be used as an important source of non-autologous NK cells; abundant cell sources can be provided for clinically using non-autologous NK cells to treat diseases such as malignant tumors and virus infection. Meanwhile, efficient amplification of the rat NK cells can be realized, and the obtained rat NK cells have the characteristics of high purity and high activity.

The present invention belongs to the field of cell culture technology, and in particular to a partitioned culture device for mouse cells. In view of the problem that errors are prone to occur when dripping nutrient solution into multiple culture dishes in the prior art, the following scheme is proposed, including a base with an opening downward and an overall barrel-shaped structure, a plurality of rectangular sliding holes distributed centripetally and in an elongated structure are opened at the top of the base near the circumferential edge, each of the rectangular sliding holes is slidably connected with a folding push rod, and a horizontal C-shaped clip with an outward opening is reserved at the top of the folding push rod; a culture dish is clamped in each of the C-shaped clips; when the present invention is used to add nutrient solution to each culture dish, it is only necessary to ensure that the liquid level in the glass capillary drops to a consistent level each time; after the liquid level drops to the lowest, the piston plate is pushed downward to raise the liquid level in the glass capillary again; and after each filling, one station is rotated again to avoid repeated filling of the culture dish.

The invention discloses a method for inhibiting an RAW264.7 cell from differentiating into an M1 cell. The method comprises the step of inhibiting the RAW264.7 cell from differentiating into the M1 cell by using miR-143-3p or a simulant thereof. According to the application, miR-143-3p is utilized to transfect and inhibit RAW 264.7 cells of mice, and the miR-143-3p targets and inhibits JAK-STAT and NF-kappa B pathways, so that the differentiation of the RAW 264.7 cells to M1 cells is inhibited, and the polarization effect of the M1 cells is remarkably inhibited.

The invention provides an application of an Angpt18 neutralizing antibody in preparation of a medicine for treating mouse cytokine storm syndrome. The research proves that the angiogenin-like protein 8 (Angpt18) can promote macrophages to release inflammatory factors such as TNF-alpha (Tumor Necrosis Factor-alpha), IL-6 (Interleukin-6), IL-1beta (Interleukin-1beta) and the like by using a molecular biology means. In the aspect of mechanism, the Angpt18 promotes polarization of M1 type macrophages and inhibits M2 type polarization of the M1 type macrophages to cause enhancement of body inflammatory factor storm, and the pathological phenomenon can be improved by knocking out the Angpt18. Therefore, the Angpt18 neutralizing antibody is synthesized and injected into a CSS mouse so as to be combined with Angpt18 in blood circulation of the CSS mouse, massive release of inflammatory factors of the organism is successfully inhibited, and the survival rate of the mouse within 72 hours is increased to 60%. The invention proves that the Angpt18 neutralizing antibody can effectively treat the mouse cytokine storm syndrome, and clarifies the molecular mechanism of the target Angpt18 of the Angpt18 neutralizing antibody for regulating the disease progress, thereby providing an effective strategy for clinically treating CSS patients.

Use of hydroxytyrosol or 3,4-dihydroxyphenylethanol-O-β-D-glucoside in promoting melanin generation. The effect of hydroxytyrosol or 3,4-dihydroxyphenylethanol-O-β-D-glucoside in the generation of melanin in recombinant human tyrosinase, mouse B16 cells, and AB-strain wild-type Danio rerio has been studied, and the results show that hydroxytyrosol or 3,4-dihydroxyphenylethanol-O-β-D-glucoside can promote the generation of melanin. Hydroxytyrosol or 3,4-dihydroxyphenylethanol-O-β-D-glucoside can be used for tanning, treating hypopigmentation diseases, blackening hair, and the like, to expand its pharmaceutical application.

The invention relates to the field of bioengineering, in particular to an artificial codon optimized nucleic acid molecule for coding red fluorescent protein. The invention provides a nucleic acid molecule for coding red fluorescent protein, the red fluorescent protein is mCherry, compared with the nucleotide sequence of the existing red fluorescent protein, the number of CpG sites of the nucleic acid molecule is increased from 56 to 92, and the content of G / C (GC3) at the third site of a codon is not less than 99%. The nucleotide sequence provided by the invention is an mCherry synonymous codon substitution sequence obtained by combining increasing the number of CpG sites and using an optimal codon strategy, and the fluorescence intensity of the synonymous codon substitution sequence in mammalian (human / mouse) cells is improved by 1.3-2.4 times compared with that of a sequence before modification; nucleic acid molecules which do not exist in the nature and can encode red fluorescent protein with higher fluorescence intensity are obtained by using a codon optimization method, and the development of living cell imaging is promoted.

The invention discloses a mouse NK cell in-vitro culture method and application, and belongs to the technical field of cell culture. The method comprises the following two culture steps: firstly, culturing for 1.5-2.5 days by using an activated culture medium containing an MEMS alpha culture medium, 15-25v / v% of FBS, beta-ME and mIL-2 (optional mIL-15 and mIL-12); and culturing for 16.5-17.5 days by using an amplification culture medium containing an MEMS alpha culture medium, FBS, beta-ME and mIL-2, wherein the cell density is controlled to be less than or equal to 2E < 6 > / mL in the whole culture process. According to the culture method provided by the invention, the amplification efficiency of the mNK cells can be remarkably improved, the initial proliferation speed of a multi-factor activation group is higher, the cells are maintained in a normal form, the transduction efficiency is improved, and sufficient high-activity cells are provided for research of the mNK cells.

The present application relates to the field of bioengineering, and particularly relates to a kind of artificial codon optimization nucleic acid molecule of the red fluorescent protein coding.The present application provides nucleic acid molecule of red fluorescent protein coding, and the red fluorescent protein is mCherry, compared with the existing red fluorescent protein nucleotide sequence, the number of CpG site is increased from 56 to 92, the content of the third position of codon G / C (GC3) is not less than 99%.The nucleotide sequence provided by the present application is the mCherry synonymous codon substitution sequence obtained by combining the increase of the number of CpG sites and the use of optimal codon strategy, which is 1.3-2.4 times higher than the fluorescence intensity of the sequence before modification in mammalian (human / mouse) cells;The present application uses the method of codon optimization to obtain the nucleic acid molecule of the red fluorescent protein that does not exist in nature, which can encode the red fluorescent protein with stronger fluorescence intensity, and promotes the development of live cell imaging.

The invention relates to the technical field of lung adenocarcinoma cell lines, in particular to a mouse Trp53 / Keap1 mutant lung adenocarcinoma cell line (Mus musculus) as well as construction and application of the mouse Trp53 / Keap1 mutant lung adenocarcinoma cell line. The mouse Trp53 / Keap1 mutant lung adenocarcinoma cell line (Mus musculus) constructed by the invention is delivered to the China Center for Type Culture Collection on May 18, 2025, and the preservation number is CCTCC (China Center for Type Culture Collection) NO: C2025163. The cell line is derived from a lung adenocarcinoma model of a C57BL / 6 strain mouse, carries the most common Keap1 mutation in lung adenocarcinoma, is the first known lung adenocarcinoma mouse cell line only carrying Keap1 and Trp53 mutations in the world at present, has the in-vivo tumorigenesis ability of the C57BL / 6 mouse, and fills the blank of the mouse mutant cell line in the level.

The invention discloses a construction method and application of an in-vitro depletion type CD8 + T cell mouse model. The construction method comprises the following steps: S1, preparing antibody coating liquid and pre-coating plate holes; s2, separating, extracting and culturing mouse CD8 + T cells; s3, carrying out induction establishment on early-stage depletion type CD8 + T cells; s4, establishment of induction of mid-term depletion type CD8 + T cells; and S5, induction establishment of the late exhaustion type CD8 + T cells. The three-high characteristics of the in-vitro depletion CD8 + T cell model, namely high phenotype fidelity, high cell survival rate and high data reproducibility, are realized for the first time, a standardized platform is provided for immune depletion mechanism research, immune checkpoint drug development and combined treatment strategy optimization, and the method has remarkable industrial application value.

The application provides an application of an Angptl8 neutralizing antibody in preparation of a drug for treating mouse cytokine storm syndrome. The present study proves that angiopoietin-like protein 8 (Angptl8) can promote macrophages to release inflammatory factors such as TNF-alpha, IL-6 and IL-1beta by using molecular biology means. In terms of mechanism, Angptl8 can cause the enhancement of the inflammatory factor storm of the body by promoting the polarization of M1 type macrophages and inhibiting the polarization of M2 type macrophages, and the knockout of Angptl8 can improve this pathological phenomenon. Therefore, we synthesize the Angptl8 neutralizing antibody and inject it into the CSS mouse to combine with the Angptl8 in the blood circulation of the mouse, successfully inhibit the large release of the inflammatory factors of the body, and improve the survival rate of the mouse within 72h to 60%. The application proves that the Angptl8 neutralizing antibody can effectively treat the mouse cytokine storm syndrome, and clarifies the molecular mechanism of the target Angptl8 in regulating the disease process, and provides an effective strategy for treating the CSS patient in the clinic.

The invention discloses resveratrol solid lipid nanoparticles and application thereof in preparation of drugs for treating hepatic fibrosis, belongs to the technical field of drugs for treating hepatic fibrosis, and explains a regulation mechanism of the resveratrol solid lipid nanoparticles for resisting rat hepatic fibrosis in vivo and in vitro. Through CCK-8, cell scratches and Transwell experiments, the influence of the novel nano dosage form on the proliferation, migration and invasion ability of rat hepatic stellate cells (HSC-T6) is detected; an Annexin V-FITC / PI and ROS kit is used for determining the influence of the nano dosage form on the HSC-T6 apoptosis, the period and the ROS content; immunofluorescence and western-blot are used for detecting the influence of a new nano dosage form on the expression content of a hepatic fibrosis specific marker of HSC, and it is found that the resveratrol solid lipid nanoparticles can effectively inhibit proliferation of rat HSC-T6 cells, induce HSC-T6 cell apoptosis, inhibit migration and invasion and regulate alpha-SMA expression. The resveratrol solid lipid nanoparticles can inhibit the activation of hepatic stellate cells and achieve the effect of resisting rat hepatic fibrosis by influencing a signal channel of TGF-beta / smad2.

The invention discloses an application of aqua glycerol channel protein AQP7 in inhibition of pancreatic beta cell ferroptosis. According to the application disclosed by the invention, the change of intracellular H2O2 amount and the change of autophagy related proteins (LC3II / I and P62) are studied by overexpressing and knocking out an AQP7 gene in a rat beta cell INS-1 by utilizing a lentivirus, the expression change condition of ferroptosis related proteins (ACSL4 and GPX4) is detected, and it is determined that AQP7 overexpression can inhibit decline of an autophagy function and occurrence of ferroptosis in the INS-1 cell under the condition of high fat and high glucose; after the AQP7 is knocked down, the autophagy of the cells is reduced, the amount of H2O2 in the cells is increased, and ferroptosis is activated. Researches show that change of AQP7 expression may become a feasible strategy for preventing beta cell ferroptosis in type 2 diabetes mellitus.

The application discloses a kind of methods, systems, equipment and media for constructing multi-species cell-integrated gene expression profile, belong to bioinformatics technical field.The method comprises: the single-cell transcriptome sequencing data of PDX model is compared with mixed genome library, and cell-gene expression profile matrix is obtained;According to the proportion of human or mouse genes, the cell is identified as human cell, mouse cell or double cell;The single-cell transcriptome sequencing data is compared with the integrated gene sequence set of human and mouse homologous genes, and the alignment result with integrated gene is obtained;Again based on the barcode of the identified human cell and mouse cell, cell-integrated gene expression profile is obtained from the obtained alignment result, and the interaction mechanism of human and mouse cells can be known further through cell clustering analysis.

The invention belongs to the technical field of medicines, and particularly relates to application of CD47 serving as a target spot to preparation of a medicine for treating sepsis. The invention discovers the application of CD47 in preparation of drugs or pharmaceutical compositions for treating sepsis-related diseases for the first time. The survival rate of a sepsis mouse can be remarkably improved by intervening CD47 expression through the CD47 monoclonal antibody / small molecule inhibitor / siRNA, and the research finds that B cell inhibition of the sepsis mouse is relieved by intervening a CD47-beta amyloid protein signal axis through the CD47 monoclonal antibody, lung and kidney injuries are relieved, and the survival rate of the sepsis mouse is remarkably improved. Therefore, the CD47 can be used as a therapeutic target for preparation of drugs for treating sepsis. A monoclonal antibody / small molecule inhibitor / siRNA aiming at CD47 can also be used as a treatment strategy of sepsis.

The present disclosure provides methods, compositions, kits, and systems useful in the determination and evaluation of the immune repertoire. In one aspect, target-specific primer panels provide for the effective amplification of nucleic acid sequences of murine T cell receptor and / or B cell receptor chains with improved sequencing accuracy and resolution over the repertoire. Variable regions associated with the immune cell receptor are resolved to effectively portray clonal diversity of a biological sample and / or differences associated with the immune cell repertoire of a biological sample.

The invention discloses an esophageal squamous carcinoma cell line and related application thereof, and relates to the field of biological medicine. According to the invention, two stable esophageal squamous carcinoma cell lines with relatively high purity are successfully established, and the preservation numbers of the esophageal squamous carcinoma cell lines are CGMCC No: 46307 and CGMCC No: 46308. The establishment of the two new esophageal squamous carcinoma cell lines makes up for the deficiency of mouse cells with sex difference of the same model in the research process of an esophageal squamous carcinoma animal model, and valuable experimental materials and tools are provided for subsequent research of esophageal squamous carcinoma. By deeply researching the biological characteristics and genetic backgrounds of the two cell lines, the pathogenesis of the esophageal squamous cell carcinoma and the effect of sex difference in the esophageal squamous cell carcinoma can be more comprehensively known, so that a new thought and a new method are provided for clinical treatment and prevention of the esophageal squamous cell carcinoma.

The invention discloses a therapeutic chimeric monoclonal antibody and application thereof. The monoclonal antibody M01 is obtained from a mouse of an immune polypeptide vaccine MAP-Loop2, and the MAP-Loop2 polypeptide vaccine is a polypeptide fragment designed on the basis of a neisseria gonorrhoeae MtrE protein Loop2 structure. All sequences of immunized mouse B cells BCR are obtained through single cell sequencing, and a variable region sequence with the highest occurrence frequency is screened and cloned to a human IgG1 antibody constant region for expression and purification and is named as M01. Researches prove that the M01 has preventive and therapeutic effects on neisseria gonorrhoeae infection in a mouse vaginal infection model. According to the invention, the chimeric antibody with the Loop2 structure and exposed on the surface of the neisseria gonorrhoeae MtrE protein is prepared, so that a complement-dependent antibody sterilization reaction is effectively initiated; neisseria gonorrhoeae infection is effectively treated, the problem of drug resistance enhancement caused by antibiotic treatment is avoided, and a new treatment means is provided for gonorrhoeae prevention and control.