543 results about "Murine model" patented technology

A transgenic non-human animal, such as a mouse, has a genome that include a nucleic acid construct having at least one transcriptional regulatory sequence capable of directing expression in B cells of the animal, wherein the transcriptional regulatory sequence is operably linked to a nucleic acid encoding a miR155 gene product. A method of testing the therapeutic efficacy of an agent in treating or preventing a lymphoproliferative condition includes assessing the effect(s) of the agent on a transgenic non-human animal.

The invention discloses a preparation method of a staphylococcus aureus CRISPR / Cas9 system and an application of the system in constructing a genetically modified mouse model. The staphylococcus aureus CRISPR / Cas9 system is composed of two components, namely Cas9 mRNA and gRNA, wherein a preparation method of the Cas9 mRNA is achieved by adding a T7 promoter to the upstream region of original Cas9 coding DNA, and a preparation method of the gRNA is achieved by adding the T7 promoter to the upstream region of an original gRNA coding sequence. The staphylococcus aureus Cas9 mRNA and gRNA, which are injected to mouse fertilized embryos through micro-injection, can achieve gene editing and modification of various types, such as single-gene knockout, multi-gene knockout and / or gene knock-in and the like, on the mouse fertilized embryos; therefore, the CRISPR / Cas9 system has a good application prospect in the aspects of fertilized embryo gene editing and modification of such animals as mouse and the like as well as construction of animal models.

The invention belongs to the technical field of bio-macromolecular pharmaceutical preparations, and specifically relates to a DNA, a plasmid and a preparation capable of directionally clearing HBV (Hepatitis B virus) ccc in hepatocyte. According to the invention, five brand-new CRISPR / Cas systems are discovered through wide and in-depth studies, and can effectively clear HBV ccc DNA in cells, inhibit reproduction of HBVs, and reduce expression of proteins related to the HBVs. A pH-sensitive and PEG-modified cationic lipid carrier is further prepared, and has the advantages of good stability and high transfection efficiency; and internal instability of multiple cationic lipid carriers is overcome. The CRISPR / Cas systems are combined with the pH-sensitive and PEG-modified cationic lipid carrier to prepare a CRISPR / Cas9 cationic lipid carrier preparation which can effectively inhibit virus reproduction in a murine model suffering from acute HBV infection and reduce antigen expression level.

Vaccines and methods of inoculation for conferring immunity to Cryptococcus infection are disclosed. Strains of Cryptococcus fungi, including Cryptococcus neoformans and Cryptococcus gattii, can be administered to a human or animal subject via inhalation. Cryptococcus fungi that can be used to confer immunity can comprise one or more mutations in genes that contribute to chitosan production, such as genes encoding a chitin deacetylase (cda), a chitin synthase (chs) and / or a regulator of chitin synthase (csr). Inhalation administration of heat-killed Cryptococcus harboring deletions in cda1, cda2 and cda3 genes can confer immunity. In a murine model system, inhalation administration of Cryptococcus neoformans harboring deletions in cda1, cda2 and cda3 genes conferred immunity against subsequent exposure to wild type Cryptococcus neoformans in 100% of test animals. Inhalation administration of heat-killed Cryptococcus grown under conditions leading to reduced chitosan production can also confer immunity.

The invention discloses a single-chain antibody of human source anti-alexin C3d molecules. A light chain and a heavy chain of the antibody have a unique CDR region; excellent antigen binding activityis realized; the affinity constant reaches 1.22*10<-7> mol / L. Biological distribution experiments prove that the anti-C3d single-chain antibody provided by the invention can be highly gathered in thearthritis positions after entering a mouse model with rheumatoid arthritis; the arthritis serious degree of the three ScFv treatment groups is obviously lower than that of a PBS group; the healing degree has the obvious dose dependency relationship; the result shows that the excellent anti-adhesion / anti-inflammatory targeted inhibition effects are achieved. In the treatment process on MRL / lpr mice with lupus erythematosus, the anti-C3d single-chain antibody provided by the invention can obviously improve the survival rate of the mice; the symptoms of proteinuria, glomerular score, interstitial inflammation, vasculitis and crescent / necrosis and the like in the treatment group are obviously relieved. The result shows that the anti C3d single-chain antibody provided by the invention has excellent application prospects in the preparation of medicine for treating autoimmune diseases.

Leptin was previously demonstrated to exert potent immune modulatory properties in several immune mediated disorders. The aim of the study was to determine leptin's anti-tumor effect in a murine model of human hepatocellular carcinoma (HCC). In vivo, Athymic T cell deficient (nude) mice transplanted with 1×106 human Hep3B cells, followed by administration of two daily intraperitoneal doses of 0.5 mg / gram leptin for 6 weeks. Leptin administration induced a significant reduction in tumor size and improved survival in nude mice. Histologically, tumors of leptin-administered mice featured increased inflammatory exudate in interphase areas. Leptin-induced tumor suppression was associated with a significant increase in peripheral natural killer (NK) cell number. Splenocytes from leptin-treated mice featured decreased expression of CIS mRNA. To determine which lymphocyte subset is a prerequisite for the anti tumor effect of leptin, T&B cell deficient (Scid) mice and T,B& NK deficient (Scid-Beige) mice were subcutaneously implanted with Hep3B tumor cells, with and without the daily intraperitoneal administration of 0.5 mg / gram leptin for 6 weeks. SCID mice featured leptin-associated tumor suppression similar to those of nude mice. In contrast, NK-deficient SCID-Beige mice developed larger tumors. To further establish natural killer cell's central role in mediation of leptin's anti-tumor effect, NK cells were incubated in vitro with increasing doses of leptin, demonstrating a dose-dependent increase in cytotoxic activity. Incubation of leptin with hepatoma cell line was found to induce a dose-dependent reduction in hepatoma cell proliferation, suggesting an additive direct anti-tumor effect. Further synergism in inhibition of hepatoma cell proliferation in vitro was achieved following addition of natural killer cells. HCC cells expressed leptin receptor mRNA, while addition of leptin induced increased mRMA expression of STAT2 and SOCS1 on tumor cell lines. Leptin administration induces a significant suppression of human HCC. This effect is mediated by induction of natural killer cell proliferation and activation, and by direct inhibition of tumor growth. Decreased natural killer cell expression of inhibitory CIS protein and over expression of the anti-proliferative STAT2 and SOCS1 proteins in HCC lines may underline both anti cancerous effects of leptin.

The invention relates to a method for building a spleen-deficiency chronic atrophic gastritis gastric precancerous lesions animal model. The method is based on the existing spleen-deficiency rat model and the existing chronic atrophic gastritis rat model and is used for building the animal model through united copy of free drinking of N-methyl-N '-nitro-N -nitrosoguanidine (MNNG) solution, irregular eating and purgation. The method comprises the following specific steps of: preparing 10mg / L MNNG storage solution by utilizing sterilized tap water, preparing 80 to 200 mug / mL MNNG solution in temporary use to substitute the drinking water to be drunk freely by the rats, and adopting the free drinking of MNNG as the beginning of the modeling; copying a spleen-deficiency syndrome through irregular eating and improved purgation way, feeding the rats on alternate days since the fifth week after the modeling, and utilizing qi-activating decoction for gavage in a dose of 1 to 3ml per rat every day in the seventh weak; and continuing the modeling for 12 weeks. The animal model built through the method is identical to or similar with the clinical syndrome and physical signs of middle-late lesions in chronic atrophic gastritis. After being verified in repeatedly modeling, the method has good repeatability and good stability. The built model is suitable for selecting traditional Chinese medicine for treating the chronic atrophic gastritis through a traditional Chinese medicine spleen treatment method.

The present invention relates to methods of treating or reducing the risk of necrotizing enterocolitis (NEC) in an infant comprising orally administering an effective amount of a CpG-ODN. It is based, at least in part, on the results of experiments in which orally administered CpG-ODNs were observed to reduce the histopathology and markers of inflammation in a murine model for NEC. The present invention further provides for oral formulations of CpG-ODN for administration to infants.

The invention discloses a rat-pig intestinal epithelial cell integrated model established by lipopolysaccharide (LPS) stimulation. Bacterial lipopolysaccharide (LPS) has a function in damaging intestinal epithelial barrier, and the integrated model disclosed by the invention aims at identifying whether an antimicrobial peptide Cathelicidin-BF (C-BF) derived from a snake has a protective effect on LPS-induced enteritidis or not. IPEC-J2 is an intestinal epithelial cell line isolated from the porcine jejunum segment and the porcine digestive system is very similar to the human digestive system. By in vivo rat model and in vitro porcine IPEC-J2 cell model, the protective effect of Cathelicidin-BF on impaired LPS-induced intestinal barrier function is tested. The test results show that the Cathelicidin-BF has protective effects on impaired LPS-induced intestinal barrier function and LPS-induced IPEC-J2 cell damage model.

The present invention discloses a preparation method and an application of a mussel oligopeptide, and belongs to the field of marine biotechnology application. The preparation method comprises: adopting mytilus edulis as a raw material, and adopting an alkaline cold extraction method to obtain mussel protein from mussel freeze-drying powder; and carrying out composite enzymolysis of protein to obtain mixed polypeptides, carrying out ultra-filtration to obtain the oligopeptide, adopting aging mouse models to carry out a series of anti-aging animal experiments and behavioral experiments, and carrying out a drosophila survive experiment to prove aging delay activity of the mussel oligopeptide so as to create a new approach for achievement of scientific and high-value mussel utilization. Compared with the method in the prior art, the method of the present invention has the following advantages that: the mussel protein is extracted and prepared, the optimal protease and the preferred process are adopted to carry out enzymolysis purification on the mussel protein, decomposition on the mussel protein molecule structure due to the high temperature is avoided, the grading of the mussel polypeptide at the molecular weight level is achieved, and the obtained mussel oligopeptide has the pure and good aging delay effect.

An effective prophylactic mucosal gene expression vaccine (GXV), made up of a cocktail of at least 4 different plasmid DNAs encoding corresponding RSV antigens, coacervated with chitosan to formulate nanospheres. In a murine model of RSV infection, intranasal administration with GXV results in significant induction of RSV-specific antibodies, nasal IgA antibodies, cytotoxic T lymphocytes, and IFN-γ production in the lung and splenocytes. A single dose of GXV induces a drastic reduction of viral titers.

The invention relates to a circulating tumor cell mouse model and a construction method and application thereof. The method includes the following steps of 1, subcutaneous transplantation, wherein a primary tumor tissue sample is transplanted into the body of an immunodeficient mouse to construct a primary cancer heterotransplantation model PDX; 2, collection of circulating tumor cells, wherein the circulating tumor cells in peripheral blood in the primary cancer heterotransplantation model obtained in the step 1 are collected; 3, renal sac membrane transplantation, wherein the circulating tumor cells collected in the step 2 are transplanted into the renal sac membrane of the immunodeficient mouse, and then the circulating tumor cell mouse model is successfully constructed. According to the method, the mode of transplantation with two or more times of passages is adopted, the CTCs in the primary cancer heterotransplantation model are transplanted into the body of the immunodeficient mouse through renal sac membrane, and the circulating tumor cell mouse model is obtained; the circulating tumor cell mouse model can be applied to research on the in-vivo metastasis mechanism and proliferation condition of CTCs.

The invention discloses an in vitro construction method of liver organs and applications. Mouse hepatocytes are extracted to induce to construct liver organs; the forms of the liver organs are observed under a microscope; and immunofluorescence and PCR identification are performed on dryness and epithelial cell properties. An acute hepatic failure mouse model after 70% hepatectomy is established;the liver functions on the first, fourth and seventh day after operation are evaluated after the liver organs are transplanted; liver-to-body ratio is compared; and the treatment effects of the liverorgans on acute hepatic failure are verified through HE staining and immunohistochemistry. Inflammations such as HE staining and ki-67 staining and proliferation indexes show that proliferation can beobviously enhanced after the liver organs are transplanted. The liver organs have strong dryness and proliferation functions; the liver functions of acute hepatic failure mice after 70% hepatectomy can be effectively relieved, and the liver-to-body ratio can be increased; and liver regeneration and repair of acute hepatic failure after 70% hepatectomy can be promoted by relieving inflammatory infiltration of livers.

The invention discloses an application of baicalin in preparation of a medicine for treating polycystic ovarian syndrome. As shown in an in vivo experiment, the baicalin has obvious treatment effect on a polycystic ovarian syndrome rat model and can greatly reduce the high androgen level of the rat model, promote follicle maturity and induce ovulation; as shown in an in vitro experiment, the baicalin can inhibit the expression of androgen of a polycystic ovarian syndrome cell model. More importantly, the baicalin has small side effect and incomparable security while having no liver and kidney toxicity. Therefore, the baicalin can be used for preparing the medicine for treating polycystic ovarian syndrome and has a brilliant prospect.

The present invention relates to methods and compositions for reducing and / or delaying one or more signs of aging which comprise inhibiting NF-kappa B activation. It is based, at least in part, on the discovery that a peptide which inhibits IKK-β interaction with NEMO, linked to a transducing peptide, inhibits the development of various indicia of senescence in a murine model of aging, Ercc1− / Δ mice.

The invention provides a construction method of a mouse model realizing conditional site-specific double-overexpression of HPV E6 / E7 genes. The construction method comprises the following steps: 1, acquiring mice: (1) respectively designing and acquiring sgRNAs; (2) respectively preparing Cas9 / sgRNA mixtures; (3) constructing a targeting vector; (4) carrying out microinjection and acquiring F0-generation mice, including F0-generation mice with HPV E6 knocked in at the specific ROSA26 site and F0-generation mice with HPVE7 knocked in at the specific H11 site; (5) acquiring F1-generation mice; and (6) acquiring F2-generation positive homozygote mice; and 2, acquiring conditional site-specific HPV E6 / E7 double-knock-in model mice: hybridizing the two F2-generation positive homozygote mice toobtain the conditional site-specific HPV E6 / E7 double-knock-in model mice. According to the method provided by the invention, the animal model which can be stably inherited and accurately regulated and controlled can be obtained.

The invention provides a construction method and an application of an HBeAg transgenic mouse model. The method comprises the following steps: CRISPR / Cas9 technology is adopted, an HBeAg gene is inserted into a pliver-HBeAg expression cassette at the Rosa26 gene locus in a fixed-point manner through homologous recombination, and a recombinant R26-e(Alb-HBeAg)1 targeting vector is constructed with an In-Fusion cloning method and contains a 3.3kb 5' homologous arm, pliver-HBeAg and a 3.3kb 3' homologous arm; Cas9mRNA, gRNA and the recombinant R26-e(Alb-HBeAg)1 targeting vector are micro-injectedinto a zygote of a C57BL / 6J mouse, then the zygote is transplanted into the uterus of a C57BL / 6J female mouse, and an HBeAg transgenic mouse is obtained through propagation and purification step by step. The transgenic mouse can specifically express HBeAg in liver, thereby being capable of applying to experimental animal models for researching HBeAg infection mechanism and evaluating treatment effects of therapeutic drugs and vaccines for chronic hepatitis B.

The invention relates to the technical field of biology, in particular to a leukemia mouse model based on a gene co-transfection technology and a preparation method thereof. The preparation method of the leukemia mouse model comprises the following steps: performing construction and packaging of K-ras mutants and AML1-ETO fusion gene lentivirus vectors; performing bone marrow cell separation and virus infection condition monitoring; implanting infection cells in a mouse to build a leukemia animal model; and performing model identification. The method of building the leukemia mouse model in a mode of utilizing caudal vein injection to lead in the manual site-directed mutagenesis K-ras mutants and AML1-ETO fusion gene lentivirus vectors is adopted initiatively. The leukemia mouse model is high in success rate, pathological characteristics are high in similarity to morbidity conditions of clinical leukemia, and the novel animal model can be provided for leukemia extramedullary infiltration mechanism research, leukemia medicine screening and gene and molecule target treatment.

The present invention discloses a fodder formulation for constructing nutritive obesity animal model, and especially fodder formulation for constructing nutritive obesity rat model. Obesity is dystrophic disease caused by the mutual action of genetic factors and environment factors and is inducement of several chronic diseases. The present inventor prepares fodder with the pre-mixed material of corn, lard, casein, flour, dried pork floss, grass powder, fish meal, bran and mineral salt, and the pre-mixed material of cholesterol and vitamins. The fodder is used in raising rat to obtain obesity model with obvious difference to the contrast in weight index and biochemical blood indexes. The animal model may be used as the platform for obesity research, medicine screening, and functional food development and identification.

The invention provides an inactivated or attenuated gene functionally linked to a hotspot for somatic hypermutation. Inventive nucleic acids include inactivated genes operatively linked to immunoglobulin gene control elements. Embodiments of the invention include murine models of plasma cell disease and germinal center cell lymphoma.

A method of preserving haptenized tumor cells is described. The method employs a freezing medium containing an effective amount of sucrose and human serum albumin in an isotonic buffered saline solution. Cryogenically preserving haptenized cells in such a medium has been found to maintain the integrity of the tumor cells during storage. The haptenized tumor cells also retain cell-associated antigens and haptens, and are as immunogenic, i.e., capable of inducing immunotherapeutic response, as fresh vaccine in a mouse model of metastatic disease. In a specific embodiment, haptenized cells are exposed to a solution of 8% sucrose, 10% human serum albumin in Hank's buffered solution, and then frozen to −80° C. overnight and then stored in a liquid nitrogen freezer. Methods of storing haptenized tumor cells and compositions are also provided.

The invention belongs to the traditional Chinese medicine field and relates to the preparation of effective parts of astragalus mongholicus and hedysarum polybotrys and selection thereof for pulmonary fibrosis model intervention so as to obtain further study on the effective parts having the optimal curative effect. An interstitial pulmonary fibrosis model of rats is established through modeling; and compared with the normal rats, the rats having interstitial pulmonary fibrosis have obvious difference and pathologic change typical cases in various related detection indexes and pathologic changes. The various effective parts of astragalus mongholicus and hedysarum polybotrys are capable of suppressing various related indexes of the interstitial pulmonary fibrosis model of rats induced by bleomycin and preventing the development of interstitial pulmonary fibrosis, wherein low and medium dose groups of hedysarum polybotrys flavone have the optimal effect. The degree of the effect of the effective parts of astragalus mongholicus and hedysarum polybotrys on stopping the progress of interstitial pulmonary fibrosis is related to appropriate concentrations of equivalent dugs. Through comprehensive assessment, the hedysarum polybotrys flavone is better in influence on the lung function, HA, LN, HYP and the like of the rats having interstitial pulmonary fibrosis than other effective parts.

The invention discloses application of recombinant human cytoglobin (rhCYGB) in preparation of drugs treating hyperlipidemia and atherosclerosis. A hyperlipidemia rat model and an atherosclerosis rat model are respectively established by the inventor, and subcutaneous injection of rhCYGB is carried out for treatment. Experimental results show that the rhCYGB drug is non-toxic, compared with a model control group, the rhCYGB treatment group has significant improvement, the four items of blood lipid test are significantly decreased than those of a model group, and the four items of blood lipid test of a polysaccharide sulfate treatment group are significantly higher than normal, thus rhCYGB has better efficacy than polysaccharide sulfate in treatment of hyperlipidemia caused by high fat diet. In the atherosclerosis rat model, compared with the model group, the four items of blood lipid test of the treatment group are also significantly reduced, and atherosclerotic plaques also obviously decreased. Therefore, treatment of hyperlipidemia and atherosclerosis by the recombinant human cytoglobin has the advantages of low toxicity, safety and effectiveness.

The invention discloses application of N6022 in treatment of aortic dissection and aortic aneurysm, and relates to the field of cardiovascular diseases. In mouse models of aortic dissection and aorticaneurysm, dilation of aorta can be inhibited after intravenous injection of the N6022 into tail veins, thereby reducing incidence and mortality of aortic dissection and aortic aneurysm; and moreover,the N6022 is capable of reducing rupture of aortic aneurysm, inhibiting occurrence of hematoma in inner walls of lumen and excessive collagen deposition on vascular walls, maintaining integrity of elastic fibers in the vascular walls, and treating the aortic dissection and the aortic aneurysm. The application of the N6022 in the treatment of the aortic dissection and the aortic aneurysm opens upa new application field of the N6022, and provides a meaningful reference for the prevention and the treatment of the aortic dissection and the aortic aneurysm diseases as well as improvement of vascular lesions.

The invention belongs to the technical field of model building and evaluating methods, particularly relates to a building and evaluating method of a chronic atrophic gastritis rat model and mainly solves the technical problem that existing building and evaluating methods of the chronic atrophic gastritis rat model are low in accuracy, high in cost and time consuming and labor consuming. Technology of metabonomics is adopted, and a metabolic profile dynamic trajectory spectrum is acquired by analyzing changes in endogenous metabolite in organism end product urine before and after model building. In addition, MestReNova software is used to process all NR spectra to obtain integral data, statistical analysis on content of 18 biomarkers is combined, and a rule that changes in integral average value of the 18 biomarkers in the urine before and after model building reflect changing tendency of urine metabolic trajectory of rats with chronic atrophic gastritis to certain extent is found, so that the chronic atrophic gastritis model is evaluated in a targeted manner.

Described herein are methods and apparatus for increasing sensitivity of ultrasound imaging of fluid flow in an object of interest. Ultrasound imaging of blood perfusion can be performed without contrast enhancement. Embodiments include transforming a spatiotemporal echo data array into a three-dimensional perfusion data array having a spatial dimension, a slow-time dimension, and a frame-time dimension, and filtering the perfusion data array with an eigen passband clutter filter. The clutter filter can increase sensitivity and utility of ultrasound imaging of fluid flow. In some aspects, the method can yield blood flow signal power and perfusion values well separated from tissue clutter. In an example, enhancements to ischemic tissue perfusion maps in a murine model are shown.

The invention relates to a novel human pancreatic cancer nude mouse model construction method for living imaging research and a use thereof. A human pancreatic cancer PANC-1 cell is stably transfected with a plasmid carrying a luc gene through a Lonza nuclear transfection system, a cell strain PANC-1-LUC for stable luciferase expression is screened through puromycin and PANC-1-LUC nude mouse transplant subcutaneous sarcoma is further constructed. An in vitro and vivo bioluminescence detection result proves that the PANC-1-LUC cell strain can stably express luciferase for a long time. Compared with a lentiviral vector-mediated bioluminescence pancreatic cancer nude mouse model, the built pancreatic cancer nude mouse model has a short tumor formation incubation period and a high tumor formation rate, satisfies PANC-1 transplantation tumor growth features and is suitable for living imaging research.

The invention discloses application of long non coding RNA (lncRNA) and interfering RNA thereof to treatment on atherosclerosis. Through high-throughput sequencing on an aortic tissue of an atherosclerosis mouse model, the invention discovers that the expression of lncRNA molecules in normal and late atherosclerosis mice is obviously different; and meanwhile, the lncRNA molecules are highly expressed in the atherosclerosis tissue and the lncRNA molecules are named as lncRNA-AFIAR. The experiment proves that the lncRNA-AFIAR has the functions of enhancing proliferation of macrophages, inhibiting macrophage apoptosis and the like and participates in the process of promoting the progress of the atherosclerosis. The aims of inhibiting the proliferation of the macrophages and reducing plaque formation can be fulfilled by inhibiting the lncRNA-AFIAR, so that the aim of inhibiting the progress of the AS. Therefore, the invention provides new molecular markers and intervention targets for diagnosing and treating the atherosclerosis and also provides new technical means for treating the atherosclerosis.

The invention relates to the technical field of biology, in particular to collagen as well as a preparation method and application thereof. The collagen provided by the invention can significantly increase skin moisture content of a D-galactose senile-induced nude mouse model; compared with a model control group, the contents of MDA in the skin and animal liver in high and medium dose groups of the collagen are obviously decreased, the content of hydro-xyproline in the skin in the high dose group of the collagen is obviously increased, the contents of SOD in the skin and animal liver in the high dose group of the collagen are obviously increased, and the content of T-AOC in animal plasma in the high dose group of the collagen is significantly increased. The test result shows that the collagen has the effects of resisting oxidation and scavenging free radicals, and has better effects on maintaining skin moisture and improving the content of the collagen in the skin.

The invention discloses a targeting vector and nucleic acid composition for constructing a liver injury mouse model, and a construction method, and relates to the technical field of genetic engineering and genetic modification. The targeting vector disclosed by the invention comprises a first expression cassette and a second expression cassette positioned at the downstream of the first expressioncassette; the first expression cassette comprises the following sequential series components: a liver-specific promoter, a tetracycline transcriptional activation regulating factor and a first polyA;and the second expression cassette comprises the following sequential series components: a second polyA, a mouse prourokinase activator coding gene, and a tetracycline-induced promoter. The liver injury mouse model constructed by the targeting vector has henotype of spontaneous liver injury and induction aggravated liver injury; the mouse death rate of the filial generation thereof is low; large-scale breeding can be conveniently carried out; and the invention provides the reliable liver injury mouse model for researching liver diseases.