44 results about "Podocyte" patented technology

The invention discloses a construction method of a COL4A5-K229X point mutation X-linked Alport syndrome mouse model. The construction method comprises the following steps: aiming at c.685Agt of a No.12 exon of a mouse COL4A5 gene; carrying out T point mutation, and designing and preparing Cas9 mRNA, gRNA and a donor vector; the components are mixed and then microinjected into fertilized eggs of a C57BL / 6J mouse to obtain an F0-generation mouse; identifying the genotype through PCR (Polymerase Chain Reaction) amplification and Sanger sequencing, and screening positive mice; mating the positive F0-generation mice with the wild-type mice, and breeding F1-generation and subsequent generations; the phenotype of the model is further verified through qPCR, biochemical analysis, light microscopic examination, transmission electron microscope and immunofluorescence. The model constructed by the invention shows hematuria, proteinuria, azemia, podocyte loss and irregular thickening and layering of glomerular basement membrane, is consistent with phenotypes of human XLAS patients, and provides an animal model tool for analyzing pathogenesis and developing treatment strategies.

The invention belongs to the field of biological medicines, and particularly discloses application of extracellular vesicles EVs carrying CD35 functional molecules in preparation of medicines for treating sepsis-related multi-organ injury, preferably, the EVs are derived from human podocytes or other cells or tool cells capable of naturally or engineered expressing CD35, and the EVs are administrated through caudal vein injection and can be used for preparing medicines for treating sepsis-related multi-organ injury. Normal saline is taken as a carrier, and the dosage of 1 * 10 < 10 > EVs each time is once every 12 hours and twice continuously. It is proved for the first time that EVs carries CD35 to inhibit circulation mononuclear cell complement activation (the EVs are combined with C3b to block downstream complement cascade reaction), migration of mononuclear cells to the kidney, the heart, the liver and the lung is reduced, and therefore multi-organ inflammation damage and cell apoptosis are relieved. In a cecum ligation perforation (CLP) sepsis model, it is proved that the medicine significantly improves the survival rate of mice, improves renal functions (reduces creatinine / urea nitrogen) and kidney pathological tissue damage, and meanwhile relieves pathological damage of multiple organs of the heart, the liver and the lung. The invention provides a new treatment strategy and drug candidate for sepsis-related multi-organ dysfunction.

The invention discloses an application of TMEM106A as a target spot in preparation of a diagnostic kit and a targeted therapeutic drug. Relates to the technical field of immunodiagnosis. Specific applications are provided. The invention discloses a key effect of TMEM106A in podocyte injury related to preeclampsia. TMEM106A is significantly down-regulated in urine of a PE patient, is closely related to podocyte injury and inflammatory response, and can be used as an early noninvasive diagnostic marker; meanwhile, the targeted podocyte up-regulation TMEM106A has remarkable anti-inflammatory and kidney protection effects, and a theoretical basis and practical feasibility are provided for targeted therapy of PE-related podocyte injury. The invention provides a new way for early precise diagnosis and intervention strategy of PE, and has wide clinical application prospect.

The invention relates to the technical field of functional foods, and particularly discloses a dietary therapy composition for promoting repair of cranial nerve damaged cells and a preparation method. The composition is prepared from the following raw materials in parts by weight: artichoke peptide, taurine, phosphatidylserine, natural nicotinic acid, guarana extract, collagen peptide, acerola cherry fruit powder, inulin, yak bone marrow powder, vitamin B family, sialic acid and sucralose. The preparation method comprises the following steps: S1, pretreating the raw materials; s2, performing nano crushing; s3, low-temperature drying; s4, embedding the inulin; and S5, mixing and forming. The artichoke peptide and the phosphatidylserine are synergistically matched, and a compound component with liver and brain axis functions is taken into consideration, so that a multi-target synergistic effect is achieved, the requirements of cell repair and neural development are synchronously met, and compared with single-component supplement or neglect of the influence of liver detoxification on brain repair in the prior art, the brain repair effect is greatly improved. The problems of low repairing efficiency and insufficient component utilization rate are solved.

This invention discloses the application of a PDLIM2 gene overexpression viral vector in the preparation of drugs for treating podocyte disease, wherein the nucleotide sequence of the PDLIM2 gene is shown in SEQ ID NO:1. This invention is the first to discover and verify the core role of the PDLIM2 gene in podocyte protection, confirming its significant downregulation in DKD and FSGS disease models, and demonstrating through functional experiments that PDLIM2 overexpression can effectively stabilize the podocyte cytoskeleton, inhibit apoptosis, and reduce proteinuria, thus establishing it as a key target gene for the treatment of podocyte disease. This invention is the first to intervene at the gene level in the core link of podocyte damage—cytoskeleton stability. By specifically upregulating the expression of PDLIM2 in podocytes, it directly enhances their intrinsic cytoskeleton support and anti-damage ability, achieving a fundamental shift in treatment strategy from "symptomatic support" to "causal repair."

The invention discloses application of an SHIP1 agonist AQX-1125 in preparation of a medicine for treating focal segmental glomerulosclerosis, and belongs to the technical field of medicines.The endogenous regulatory factor SHIP1 agonist is used for conducting precise speed limiting on a PI3K / Akt pathway, so that a new effective means is provided for FSGS treatment, and the application form of the SHIP1 agonist AQX-1125 can be an oral preparation and the like; tests prove that the AQX-1125 can remarkably repair renal functions, reduce pathological proteinuria, improve pathological damage of kidney tissues, maintain phenotypic homeostasis and ultrastructural integrity of podocyte and accurately regulate and control pathogenic signal pathways and metabolic homeostasis, so that the AQX-1125 shows remarkable technical progress and clinical application potential in the aspect of FSGS treatment.

A viral vector, wherein the viral vector comprises a COL4A3, COL4A4 or COL4A5 transgene.

The invention belongs to the technical field of biological medicine, and discloses a construction method and application of a podocyte specific S1PR1 gene knockout mouse model. A podocyte specific S1PR1 knockout mouse is constructed through a Cre-LoxP system, and then an FSGS model is constructed through doxorubicin injection. And verifying the model by using urine detection, serum detection, histopathology detection and molecular mechanism detection. Research finds that S1PR1 expression decline causes POLR2A splicing abnormity and expression decline through down-regulation of Xab2, and podocyte senescence and FSGS progress are induced. According to the invention, a related model is constructed for the first time, the regulation effect of the S1PR1-Xab2-POLR2A pathway in podocyte senescence is disclosed, a reliable tool is provided for researching FSGS pathogenesis, the potential of S1PR1 as a therapeutic target is determined, and the S1PR1 can be used for screening drugs for treating FSGS and has important clinical transformation value.

The application provides application of a TAS2R4 agonist in preparation of a drug for treating and preventing diabetic nephropathy, and comprises that a bitter taste receptor 4 subtype (TAS2R4) can be used as a drug target point for treating and preventing diabetic nephropathy, and quinine is used as a TAS2R4 agonist. Specifically, mouse diabetic nephropathy induced by a chemical reagent and mouse podocyte cell line MPC cell damage caused by chronic high glucose are taken as research objects, it is found that mouse TAS2R4 agonist quinine has a prevention and treatment effect on diabetic nephropathy, has a protection effect on podocyte loss caused by high glucose, and has an activation effect on mouse kidney TAS2R4 molecular signal, and a possible molecular mechanism is discussed from the aspects of inhibiting NLRP3 inflammasome activation and NF-kappa B signal activation path, and theoretical basis and technical support are provided for prevention and treatment of diabetic nephropathy, other kidney diseases with inactivation of TAS2R4 molecular signal and other diseases by using TAS2R4 agonists including quinine.

The invention relates to an application of a reagent for detecting SRC expression in preparation of a product for diagnosing and treating lupus nephritis, SRC is highly expressed in a patient with lupus nephritis, when the expression level of SRC is reduced, ferroptosis of podocyte is remarkably relieved, ferroptosis related markers are remarkably reversed, the levels of Fe < 2 + > and MDA are reduced, the level of GSH is increased, and compared with a control group, the SRC expression level is remarkably reduced. The viability of podocyte is increased; the invention relates to application of a reagent for detecting CYBB expression in preparation of a product for diagnosing and treating lupus nephritis, CYBB is highly expressed in a patient suffering from lupus nephritis, when the expression level of CYBB is reduced, ferroptosis in podocytes is remarkably relieved, ferroptosis related markers are remarkably reversed, the levels of ACSL4, Fe < 2 + >, ROS and MDA are reduced, and the levels of GPX4 and GSH are increased, so that the effect of treating lupus nephritis is achieved. The polarization of M2 macrophages is promoted, the ferroptosis phenomenon is further relieved through the polarization of the macrophages, and the viability of podocytes is improved.

The invention discloses a Chinese catalpa fruit-based unilateral preparation for eliminating urine protein and application thereof, and belongs to the technical field of traditional Chinese medicine, the active ingredient of the unilateral preparation is extracted from a single medicinal material of Chinese catalpa fruit, and the unilateral preparation can contain pharmaceutically acceptable auxiliary materials. The single preparation can be prepared into four dosage forms of oral liquid, tablets, dripping pills and granules so as to meet different clinical requirements. Animal experiments prove that the single preparation can effectively eliminate urine protein and improve the renal function through four synergistic mechanisms of anti-inflammation, anti-oxidation, improvement of the renal function, protection of podocyte and immunoregulation and diuresis, and is suitable for treating proteinuria accompanied by nephropathy. The preparation provided by the invention has the characteristics of single formula, easy quality control and various dosage forms, and provides a traditional Chinese medicine scheme with an exact curative effect for clinically treating related diseases.

The present application relates to a kind of tumor infiltrating lymphocyte (TIL) cell expansion culture process. Specifically, the present application provides a kind of TIL cell expansion method, comprising: in expansion culture medium, preliminary culture TIL cell is carried out fast expansion culture. In the TIL cell expansion method of the present application, it is not dependent on activation antibody CD3 antibody, high-dose IL-2 and feeder cell, that is, the TIL cell number hundreds of times expansion can be realized in short time, the demand of TIL cell quantity in TIL cell therapy clinically can be satisfied, and it has broad application prospect in the field of TIL cell treatment.

A low-salt fasting-mimicking diet (LS-FMD) is provided for promoting kidney regeneration and treating kidney disease. The LS-FMD comprises cyclic dietary interventions that reduce caloric and sodium intake to induce fasting-like metabolic effects while maintaining essential nutrition. In animal models of puromycin-induced nephrosis, LS-FMD cycles restore normal proteinuria levels and improve nephron structure and function relative to untreated controls. The diet activates kidney-specific gene expression programs and promotes a quiescent, non-proliferative state in mature podocytes, supporting renal recovery. In human subjects with chronic kidney disease, administration of fasting-mimicking diet cycles for five days per month over three months reduces proteinuria and improves endothelial function. The invention demonstrates that kidney-specific fasting-mimicking dietary cycles have therapeutic potential for slowing or reversing progressive renal disorders.

The invention discloses a monoterpene indole alkaloid compound as well as a preparation method and application thereof, and belongs to the technical field of medicines. The monoterpene indole alkaloid compound disclosed by the invention is separated from ervatamia palmata, has remarkable activity for repairing podocyte injury under the hypoxia condition, and can be used for preparing a medicine for treating chronic renal injury under the hypoxia condition. The traditional Chinese medicine composition has relatively high economic benefits aiming at the current situation of non-hypoxia chronic kidney injury specific medicines.

The invention relates to the technical field of biomedical engineering, in particular to a culture medium and a culture method of podocyte fate organoid. Compared with human induced pluripotent stem cells (hiPSCs), cells separated from human adult kidney tissues are taken as objects to induce and culture podocyte fate organoid, so that the podocyte fate organoid has better gene stability and can better reproduce original tissue phenotypes. Compared with a hiPSCs-derived kidney organ, the culture method of the podocyte fate organ provided by the invention has the advantages that a culture system is simplified; the serum-free culture medium component based on a pure compound combination also greatly improves the safety of a culture product possibly used for clinical biological treatment.

The application discloses a method for extracting mitochondria-associated membranes (MAM) and other subcellular components from podocytes, which comprises the following steps: pretreating the podocytes, crushing the podocytes by low-power ultrasonic, separating the crude mitochondrial components by differential centrifugation, and then purifying the high-purity MAM and other subcellular components by density gradient centrifugation and ultracentrifugation. The method replaces the traditional homogenizer grinding with low-power ultrasonic crushing, improves the MAM extraction yield by controlling the ultrasonic parameters, appropriately simplifies the differential centrifugation steps, increases the yield while ensuring the purity, sequentially separates the unbroken cells, the crude mitochondria, the cytoplasm and the endoplasmic reticulum components, and finally obtains the high-purity MAM and other subcellular components by density gradient centrifugation and ultracentrifugation purification. The method effectively improves the yield of the podocyte separation and extraction of the MAM and other subcellular components, reduces the cost, and provides technical support for the research on the podocyte MAM proteomics.

This invention belongs to the field of biomedical technology and discloses a method for constructing and applying a podocyte-specific S1PR1 gene knockout mouse model. Podocyte-specific S1PR1 knockout mice were constructed using the Cre-LoxP system, and then an FSGS model was established via doxorubicin injection. The model was validated using urine, serum, histopathological, and molecular mechanism detection methods. The study found that decreased S1PR1 expression induces podocyte senescence and FSGS progression by downregulating Xab2, leading to abnormal POLR2A splicing and reduced expression. This invention is the first to construct such a model, revealing the regulatory role of the S1PR1-Xab2-POLR2A pathway in podocyte senescence, providing a reliable tool for studying the pathogenesis of FSGS, clarifying the potential of S1PR1 as a therapeutic target, and enabling drug screening for FSGS treatment. It has significant clinical translational value.

Methods are provided for delaying onset or progression of kidney damage, or treating kidney disease, in a subject who has type 1 diabetes or type 2 diabetes or glomerular disease. The methods comprise administering to a subject with type 1 or type 2 diabetes or glomerular disease an effective amount of an agent capable of inhibiting NBL1 activity, and in particular, capable of inhibiting NBL1-mediated toxicity of human podocytes. In some embodiments, the agent is an antibody capable of binding to human NBL1.

The invention provides a Col4a5 + / -mouse immortalized podocyte line, and the preservation number of the Col4a5 + / -mouse immortalized podocyte line is CGMCC (China General Microbiological Culture Collection Center) NO.46563. The cell line has relatively strong stability guarantee and can be subjected to continuous passage for 30 generations. The Col4a5 < + / -> mouse immortalized podocyte line corresponds to the phenotype of a female XLAS carrier with a light disease degree, can be used for precise pathological modeling, realizes COL4A5 gene dose effect simulation, and can be used as a cell model for screening a treatment drug or a diagnosis drug for hereditary nephropathy. The cell line is subjected to immortalization treatment, can be massively cultured and subcultured, has relatively simple culture requirements, can obtain a large number of experimental cells in a short time, has a short total experimental period, is suitable for popularization, and overcomes the technical problems of difficulty in constructing a single allele mutation (Col4a5 + / -) cell line, low transfection efficiency, allele loss and the like in the prior art.

Provided are dosing regimens for the treatment of Fabry disease in a patient. Certain methods relate to the treatment of ERT-experienced or ERT-naïve Fabry patients. Certain methods comprise administering to the patient about 123 mg free base equivalent of migalastat for improving left ventricular mass and / or improving podocyte globotriaosylceramide.

This invention discloses Astragalus polysaccharide, extracted from Astragalus membranaceus. The average molecular weight of this Astragalus polysaccharide is 8-12 kDa, and its monosaccharides are mainly composed of galacturonic acid, glucose, galactose, and arabinose. This Astragalus polysaccharide is a GalA-type acidic polysaccharide that effectively reduces the deposition of immune complexes and the release of pro-inflammatory factors in renal tissue by regulating signaling pathways such as cGAS-STING. Simultaneously, it improves glomerular filtration function by stabilizing the podocyte cytoskeleton structure, exhibiting a synergistic protective therapeutic advantage. This invention also discloses a method for preparing this Astragalus polysaccharide.

The invention provides application of miR-181c-5p and an extracellular vesicle containing the miR-181c-5p in preparation of a medicine for treating diabetic nephropathy, and belongs to the technical field of biological medicine. Researches find that miR-181c-5p and extracellular vesicles containing miR-181c-5p can inhibit intracellular Fe < 2 + > generation, ROS generation, MDA accumulation and increase GSH synthesis by regulating and controlling expression of TFRC genes, ACSL4 genes and PTGS2 genes, and then podocyte ferroptosis is inhibited, so that the effect of improving glomerular injury in the diabetic nephropathy process is achieved; the miR-181c-5p and the extracellular vesicles containing the miR-181c-5p may be key targets for treating the diabetic nephropathy, can be used for developing medicines for treating the diabetic nephropathy, and have good practicability.

The invention discloses a biomarker for predicting the curative effect of fenerenone on treating diabetic nephropathy and application of the biomarker, and four biomarker combinations with clear prediction values are screened and verified on the basis of a core molecular mechanism (mTOR / ULK1 pathway-podocyte autophagy) of fenerenone for treating diabetic nephropathy. And a'baseline + early stage 'dual pre-judgment system and a standardized application process are established. The scheme solves the bottleneck that the curative effect of the fenerenone cannot be pre-judged in the prior art, has the advantages of being clear in mechanism, convenient to detect and accurate in prediction, can be directly applied to clinical individualized treatment guidance, improves the treatment effective rate and reduces the medical cost, and has wide industrialization prospects.

The invention relates to the technical field of sample preservation. The invention provides an exfoliated cell preserving fluid and a preparation method and application thereof, the preserving fluid comprises a cracking agent, a buffering agent, a chelating agent, a mucus dissolving agent and a preservative, the cracking agent and the buffering agent comprise ammonium salt, and the ammonium salt comprises at least one selected from the group consisting of ammonium chloride, ammonium acetate and ammonium bicarbonate. The preserving fluid can selectively lyse red blood cells to reduce blood background interference, and strongly disperse and dissolve mucus to effectively release wrapped target cells; the morphological integrity and nucleic acid integrity of nucleated cells are protected, and morphological abnormalities such as solid shrinkage, dissolution or expansion of the cells are avoided. The biological stability of the sample in the long-term storage and transportation process is ensured due to strong anti-corrosion and antibacterial capabilities; the preservation solution can meet the requirements of cell morphology examination and molecular detection at the same time, double-detection compatibility is achieved, the preparation technology is simple and convenient, and large-scale production is easy to achieve.

This invention discloses a method for constructing a mouse model of COL4A5-K229X point mutation X-linked Alport syndrome. Cas9 mRNA, gRNA, and a donor vector were designed and prepared targeting the c.685A>T point mutation in exon 12 of the mouse COL4A5 gene. These components were mixed and microinjected into C57BL / 6J mouse zygotes to obtain F0 generation mice. Genotypes were identified by PCR amplification and Sanger sequencing, and positive mice were screened. Positive F0 generation mice were mated with wild-type mice to breed F1 and subsequent generations. The model phenotype was further verified by qPCR, biochemical analysis, light microscopy, transmission electron microscopy, and immunofluorescence. The model constructed by this invention exhibits hematuria, proteinuria, azotemia, podocyte loss, and irregular thickening and stratification of the glomerular basement membrane, consistent with the phenotype of human XLAS patients, providing an animal model tool for elucidating the pathogenesis and developing treatment strategies.

The invention discloses a traditional Chinese medicine composition for reducing urine protein and a preparation method of the traditional Chinese medicine composition, and belongs to the technical field of traditional Chinese medicine. The traditional Chinese medicine composition is prepared from astragalus membranaceus, caulis sinomenii, kadsura pepper stem, rhizoma dioscoreae nipponicae, abelmoschus manihot flower, gordon euryale seed and ligusticum wallichii according to a specific proportion; a double-vine-one-dragon wind-dispelling structure synergistically resists inflammation, enhances the penetrability of kidney collaterals and improves the antibody clearance rate, radix astragali-gordon euryale seeds synergistically protect podocytes, three-dimensional synergy of dispelling wind, dredging collaterals, immobilizing and absorbing essence, removing dampness and detoxifying is formed, a closed loop of reducing leakage, enhancing retardation and promoting repair is formed, clinical and animal experiments show that the urine protein reduction rate in 24 hours exceeds 60%, and the effect is remarkable. The effective rate is 80%, immune complex deposition can be synchronously reduced, podocyte can be repaired, the function of renal tubules can be improved, and an efficient and safe solution is provided for increasing of urine protein of multiple types of nephropathy.

Methods are provided for delaying onset or progression of kidney damage, or treating kidney disease, in a subject who has type 1 diabetes or type 2 diabetes or glomerular disease. The methods comprise administering to a subject with type 1 or type 2 diabetes or glomerular disease an effective amount of an agent capable of inhibiting NBL1 activity, and in particular, capable of inhibiting NBL1-mediated toxicity of human podocytes. In some embodiments, the agent is an antibody capable of binding to human NBL1. Monoclonal anti-NBL1 antibodies are also provided.

The invention belongs to the technical field of biological medicines, and particularly relates to application of a NOTCH1 inhibitor or a compound in resisting diabetic nephropathy. The NOTCH1 inhibitor is a reagent for inhibiting the expression of NOTCH1 or a reagent for inhibiting the expression of an ANK structural domain of NOTCH1; the compound is prepared from any one or more of 2 ', 4', 5, 7-tetrahydroxyflavanone, icariin A, cassythine, anonaine, tenuigenin, bergamotin, teasel root saponin VI, (+) magnoline and piperlongumine. The invention discloses a new mechanism that NOTCH1 is directly recruited through an intracellular structural domain of NOTCH1 and is combined with KEAP1 to promote proteasome-dependent NRF2 polyubiquitination degradation so as to regulate and control oxidative damage and ferroptosis of podocytes. Meanwhile, it is found that all the nine compounds can inhibit a NOTCH1 signal channel and have the potential of treating DKD through a targeted NOTCH1 / KEAP1 / NRF2 axis.

According to the lipid metabolism pathway marker of GK in obesity-related glomerulonephritis podocyte damage and the diagnosis application, the expression level of glycerol kinase in a tested sample is detected, and the expression level is compared with the corresponding expression level in a reference sample; diagnosis information for representing the abnormal state of the podocyte lipid metabolism pathway of the obesity-related glomerulonephritis is obtained based on the comparison result, and the expression level comprises the messenger ribonucleic acid expression level of glycerol kinase, the protein expression level or the combination of the messenger ribonucleic acid expression level and the protein expression level. A test sample comprises at least one of a kidney tissue sample, a serum sample, a plasma sample, a whole blood sample or a urine sample, and the related state of podocyte damage of obesity-related glomerulonephritis is subjected to molecular level characterization from the lipid metabolism pathway level; the method has the advantages of being clear in diagnosis directivity, suitable for multiple sample types, flexible in detection mode and capable of being used for disease layering and dynamic evaluation.