38 results about "Interstitial cell" patented technology

The application discloses a kind of hepatic organoids containing mesenchyme induction method, belong to organoid culture technical field.The method includes the following steps: (1) human embryonic stem cell WIBR3 pretreatment;(2) utilize human embryonic stem cell WIBR3 and induce liver organoids and mesenchymal cells;(3) mesenchymal cells and liver organoid cells are co-cultured, i.e. obtain the hepatic organoids containing mesenchyme.There is beneficial effect: the application changes the culture mode and method of the hepatic organoids containing mesenchyme.The hepatic organoids containing mesenchyme induction method proposed in the application is simple, and mesenchymal cells and other liver cells have the same genetic background source, suitable for the safety evaluation of drug and other chemical substances, liver fibrosis, non-alcoholic hepatitis modeling and pharmacodynamic evaluation.

The invention relates to the technical field of biological medicine, and discloses rapamycin-loaded microemulsion gel as well as a preparation method and application thereof. The preparation method of the microemulsion gel comprises the following steps: dissolving N, N-dimethyl octadecyl carboxymethyl chitosan in a water phase, then adding an oil phase, a cosurfactant and rapamycin, and stirring to obtain rapamycin-loaded microemulsion; the preparation method comprises the following steps: dissolving a high polymer material in a PBS solution, heating until the high polymer material is completely dissolved, then adding dopamine, stirring for reaction, and finally dialyzing and freeze-drying to obtain the adhesive. And adding a gel matrix containing a thickening agent and an adhesive into the microemulsion, and stirring to obtain the rapamycin-loaded microemulsion gel. According to the prepared microemulsion gel, transdermal drug delivery of hydrophobic drugs is achieved, testis are coated with the microemulsion gel loaded with rapamycin, the blocking effect of damp-heat stress on differentiation and development of testis interstitial cells can be relieved, and the functionality of a male reproduction endocrine system is recovered.

The application provides a WNT5B-promoting tooth differentiation biological product and an application thereof. Compared with the prior art, the application has the following advantages: the patent integrates human embryonic tooth germ spatial transcriptome sequencing and epithelial cell and interstitial cell extracellular protein group sequencing data, screens signal molecules WNT5B and CTNNB1 secreted in the development process of tooth germ epithelium, and confirms the important role of WNT5B protein in tooth germ development for the first time. Compared with other WNT family members, WNT5B has stronger tooth-forming ability of promoting tooth-derived stem cells; the effective concentration of WNT5B is low, and 10 ng / ml is the optimal concentration of WNT5B for promoting tooth-derived stem cells to form teeth; and the role of WNT5B and CTNNB1 in extracellular secretion of tooth germ epithelial cells and the synergistic effect of WNT5B and CTNNB1 in promoting tooth differentiation of tooth germ mesenchymal cells are further determined, that is, the combined low-concentration use effect of the two is better than the single high-concentration use.

The utility model relates to a kind of human adenomyosis ectopic epithelium and stromal cell separation culture devices. The device includes L-shaped base, fixed tube, centrifugal tube, filter separation component. Two centrifugal tubes are used to collect stromal cell and epithelial cell respectively, and the top end of centrifugal tube is connected with top cover. Filter separation component includes mounting ring, locating ring, flexible filter screen, annular frame and positioning mechanism. Mounting ring is horizontally arranged, locating ring is peripherally arranged on mounting ring, and locating ring is adapted to the top end of centrifugal tube, filter screen is arranged at the bottom of mounting ring, the bottom of annular frame is connected with the bottom surface of flexible filter screen, gap is provided between the top of annular frame and mounting ring, and annular frame is limited by positioning mechanism. Compared with prior art, the utility model can simplify the separation process of human adenomyosis ectopic epithelium and stromal cell, reduce cleaning work, and is suitable for laboratory culture operation.

The invention belongs to the technical field of biological medicine research, and particularly relates to a cell in-vitro calcification inducer, an induction culture medium and application thereof, an aortic valve calcification in-vitro induction model and a modeling method thereof. The inducer provided by the invention is prepared from the following components in parts by weight: 2160.4 parts of beta-sodium glycerophosphate, 0.039 parts of dexamethasone, 50 parts of vitamin C and 34 parts of vitamin D. The invention further provides a culture medium formed by using the inducer and a method for modeling an aortic valve calcification in-vitro induction model by using the inducer. According to the method, osteogenic differentiation of the aortic valve interstitial cells can be stably induced within a short time, an efficient and repeatable standardized induction scheme is provided for in-vitro aortic valve calcification models and drug screening, and the method has a very good application prospect.

The invention belongs to the field of biological medicine, and relates to application of triptolide in preparation of a medicine for preventing and treating calcified aortic valve diseases. The triptolide is found to be capable of promoting expression and maturation of CTSD and activating enzyme activity of the CTSD, meanwhile, biogenesis of lysosome in valvular interstitial cells can be promoted by regulating and controlling AKT / mTOR / TFEB signal axes, lysosome functions are improved, autophagy flux is recovered, and finally the effect of treating the calcified aortic valve disease is achieved. A novel treatment medicine is provided for the calcified aortic valve disease.

This invention belongs to the field of biomedical technology and relates to the application of OSM inhibitors in the preparation of drugs for the prevention or treatment of calcific aortic valve disease. This invention reveals for the first time that OSM drives osteogenic differentiation and calcified nodule formation of valvular interstitial cells through the OSM-OSMR-JAK2 / JAK3-STAT3-RUNX2 signaling axis, which is the core pathogenic mechanism leading to calcific aortic valve disease. This invention utilizes anti-OSM neutralizing antibodies or sulforaphane to effectively block this signaling axis, significantly inhibiting macrophage infiltration, valvular calcification, and improving hemodynamic abnormalities. This invention provides a precise target and a novel strategy for the modification therapy of calcific aortic valve disease, solving the technical problem of the ineffectiveness of traditional broad-spectrum anti-inflammatory therapy in valvular calcification.

The invention relates to the technical field of genetic engineering, in particular to application of PCK1 in preparation of products for regulating and controlling goat testicular interstitial cell proliferation. According to the invention, an over-expression plasmid pcDNA3.1 (+)-PCK1 of the PCK1 gene is constructed, an interference sequence si-PCK1 of the PCK1 gene is synthesized, and it is proposed for the first time that the over-expression of the PCK1 gene can significantly promote the proliferation of goat testicular interstitial cells, and the inhibition of the expression of the PCK1 gene can significantly inhibit the proliferation of the goat testicular interstitial cells. The invention has important significance for improving the development of the goat testis, and provides technical support for researching and developing related products for regulating and controlling the proliferation of the goat testis interstitial cells by taking the PCK1 gene as the target spot.

The invention provides application of disulfiram in preparation of a medicine for treating calcified aortic valve diseases, and belongs to the field of biological medicine. The disulfiram is used for intervening in-vitro cultured valvular interstitial cells and a mouse in-vivo calcified aortic valve stenosis model, and experimental results show that the disulfiram can inhibit the capability of differentiating the valvular interstitial cells to osteogenic phenotypes, relieve extracellular calcification deposition and relieve aortic valve calcification and stenosis. Therefore, the disulfiram can be used as a medicine for treating the calcified aortic valve disease and has a wide application prospect.

The invention discloses application of H-151 in preparation of drugs for preventing and treating calcified aortic valve diseases, relates to the technical field of biological medicines, and is characterized in that the invention provides application of an STING inhibitor H-151 or pharmaceutically acceptable salts thereof in preparation of drugs for preventing and / or treating calcified aortic valve diseases. The invention reveals for the first time that H-151 can inhibit the increase of the expression level of osteogenic differentiation markers ALP and RUNX2 in aortic valve interstitial cells induced by an osteogenic culture medium in an in-vitro experiment, and inhibit osteogenic differentiation and calcium salt deposition of the aortic valve interstitial cells; in addition, aortic valve thickening, fibrosis and calcium salt deposition of mice induced by an aortic valve injury model are inhibited in vivo. Based on the biological effect, the application of H-151 in developing the medicine for treating the calcified aortic valve disease is provided for the first time, and a new technical path and a new medicine choice are provided for preventing and treating the calcified aortic valve disease.

The invention discloses an establishment method and application of an iPSCs (induced pluripotent stem cells)-derived valvular interstitial cell calcification model, and belongs to the technical field of cell model establishment. The construction method of the iPSCs-derived valvular interstitial cell calcification model comprises the following steps: (1) carrying out induced differentiation for 7 days by using the combination of four regulatory factors, namely BMP4, CHIR99021, VEGF and Forskolin, so as to differentiate mature valvular interstitial cells; and (2) culturing the iPSCs-derived valvular interstitial cells in the step (1) in an osteogenic induction culture medium at 37 DEG C under the condition of 5% CO2 for at least 3 days. The invention provides a rapid and efficient differentiation scheme, the mature valvular interstitial cells induced by the scheme are very close to human primary valvular interstitial cells, and a high-quality experimental platform is provided for screening and mechanism research of anti-calcification drugs.

The application discloses a device for predicting the recurrence / metastasis risk of a non-small cell lung cancer patient after operation and application thereof, adopts multiple machine learning algorithms to combine the clinical characteristics, genomic sequencing data and immune microenvironment infiltration characteristics of real world non-small cell lung cancer patients with disease-free survival, and constructs a postoperative recurrence prediction model. Through a Lasso coefficient path diagram, it is confirmed that five characteristics including tumor size, tumor interstitium CD8+ T cell positive rate, tumor parenchyma M2 type tumor-related macrophage positive rate, TP53 gene mutation state and postoperative ctDNA-MRD state are included in model construction, and a MIMIR prediction model is constructed through a random survival forest. The model can help clinicians to accurately evaluate the postoperative disease-free survival of non-small cell lung cancer patients, guide the development of individualized treatment and follow-up schemes, and bring better survival benefits to patients.

The invention relates to the technical field of genetic engineering, in particular to application of miR-145-3p as a target spot in preparation of products for regulating and controlling proliferation of goat testicular interstitial cells. According to the invention, Inhibitor and Mimics sequences taking the miR-145-3p gene as a target spot are constructed, and it is proposed for the first time that overexpression of the miR-145-3p gene can significantly inhibit proliferation of goat testicular interstitial cells, and inhibition of the miR-145-3p gene can significantly promote proliferation of goat testicular interstitial cells. The invention has important significance for improving the development of the goat testis, and provides technical support for researching and developing related products for regulating and controlling the proliferation of the goat testis interstitial cells by taking the miR-145-3p gene as a target spot.

The invention belongs to the technical field of reproductive biology, and particularly provides a method for separating and culturing testicular perichondrial muscle-like cells of an adult mouse. The method comprises the following steps: removing interstitial cells from testis tissues of an adult mouse, and digesting the testis tissues by pancreatin and DNA enzyme to obtain a testis single-cell suspension; and culturing by adopting a new isolated culture medium until the culture dish is full of cells, and carrying out cell passage, cell cryopreservation and resuscitation and cell identification by adopting passage culture mediums with different concentrations. According to the method for separating and culturing the mouse peridermoid cells, the cell separation degree is far higher than that in the prior art, operation is easy, time is saved, the technical blank that the mouse peridermoid cells are difficult to separate and culture is filled, and the method is suitable for large-scale popularization and application. And convenience is provided for research on interaction of the mouse testicular peritubular muscle-like cells with other testicular somatic cells and germ cells.

This invention provides the application of costunolide in the preparation of a drug for preventing posterior capsule opacification. Costunolide effectively reduces posterior capsule opacification after extracapsular lens extraction (ECLE) and exhibits significant anti-inflammatory and anti-fibrotic effects in both in vitro and in vivo models. Mechanistic studies have identified matrix metalloproteinase 8 (MMP8) as a key downstream effector of costunolide, confirming that costunolide inhibits MMP8 expression, thereby blocking the transformation of lens epithelial cells (LECs) into mesenchymal cells and subsequent fibrotic processes. This lays a solid foundation for developing targeted therapies for posterior cataract, a common surgical complication.

The application discloses a method for synchronously marking germ cells and interstitial cells in the testis tissue of Paralichthys olivaceus, which comprises the following steps: cloning MIR17HG-a (long non-coding RNA) of Paralichthys olivaceus by using a probe primer, obtaining a target plasmid after plasmid extraction, carrying out double enzyme digestion and purification to obtain an enzyme-digested plasmid as a probe template, synthesizing Paralichthys olivaceus MIR17HG-a probe by using an SP6 / T7 RNA polymerase, preparing a tissue slice from the testis tissue of Paralichthys olivaceus, and carrying out fluorescent in situ hybridization on the tissue slice by using the Paralichthys olivaceus MIR17HG-a probe, so that the Paralichthys olivaceus MIR17HG-a probe can synchronously mark the germ cells and interstitial cells in the testis tissue of Paralichthys olivaceus. The method solves the defects of traditional marking technology, such as the need for combined marking or multiple step-by-step marking, and fills the blank of synchronous marking of germ cells and interstitial cells in the testis tissue of Paralichthys olivaceus.

The invention provides a separation method and an identification method of bactrian camel testis interstitial cells, and belongs to the technical field of cell separation. The method comprises the following steps: mixing 0.08-0.12% of collagenase IV solution and 0.23-0.27% of trypsin-EDTA (Ethylene Diamine Tetraacetic Acid) according to a volume ratio of 1: 2, digesting a bactrian camel testis tissue block, then carrying out hypotonic treatment purification to remove redundant germ cells, carrying out differential adhesion purification to remove a large amount of supporting cells, carrying out percoll density gradient centrifugation purification to remove a small amount of supporting cells, and carrying out freeze drying to obtain a finished product. And transferring the suspension with the percoll gradient of 34-60% into a new culture bottle to obtain the interstitial cells with the purity of more than or equal to 95%, and completing identification by combining morphological observation, immunofluorescence staining and secretion function detection. The separated bactrian camel testis interstitial cells can be used for double-hump reproductive performance research, and a foundation is laid for the reproductive performance of bactrian camels.

The invention discloses a method for synchronously marking germ cells and interstitial cells in paralichthys olivaceus testis tissues, which comprises the following steps: cloning paralichthys olivaceus MIR17HG-a (long non-coding RNA) by using a probe primer, and extracting plasmids to obtain target plasmids; the method comprises the following steps of: synthesizing a paralichthys olivaceus MIR17HG-a probe through SP6 / T7 RNA polymerase, preparing paralichthys olivaceus testis tissues into tissue slices, and performing fluorescence in-situ hybridization on the tissue slices by using the paralichthys olivaceus MIR17HG-a probe, so that the paralichthys olivaceus MIR17HG-a probe can synchronously mark germ cells and interstitial cells in the paralichthys olivaceus testis tissues. The defect that a traditional marking technology needs combined marking or multi-time step-by-step marking is overcome, and the blank of synchronous marking of germ cells and interstitial cells in paralichthys olivaceus testis tissue at present is filled.

The application belongs to the technical field of medicine, and particularly relates to a use of a flavonoid compound in aortic valve disease or cardiovascular aging. The flavonoid compound shown in formula I is found to be capable of inhibiting the osteogenic differentiation of valve interstitial cells for the first time. In animal implementation, the flavonoid compound can significantly reduce the peak flow rate and peak pressure difference of the aortic valve of a calcified aortic valve mouse, and has a significant effect of relieving aortic valve calcification. Therefore, the flavonoid compound has a good application prospect in the preparation of a drug for relieving aortic valve disease, relieving cardiovascular aging and other related cardiovascular diseases.

The invention relates to the technical field of probiotics, in particular to a lactic acid bacteria mixture for preventing and delaying heart valve calcification and application of the lactic acid bacteria mixture, and the lactic acid bacteria mixture comprises a saliva combined lactobacillus BR202 strain, a cheese paracasei BR176 strain, a bifidobacterium animalis subsp. Lactis BR621 strain and a plant lactobacillus PQQ-1 strain. According to the invention, human valvular interstitial cells are induced to generate osteogenic differentiation, so as to simulate pathological changes in calcified aortic valve diseases for evaluation, and the results of calcium salt deposition of the human valvular interstitial cells and osteogenic differentiation experiments of the human valvular interstitial cells prove that the human valvular interstitial cells can be used for simulating the pathological changes in the calcified aortic valve diseases. The lactic acid bacteria mixture can effectively delay calcium salt deposition of human valvular interstitial cells and osteogenic differentiation of the human valvular interstitial cells, so that the lactic acid bacteria mixture has the physiological activity function of preventing and delaying cardiac valve calcification, and can be used for preparing medicines or food compositions for preventing and delaying cardiac valve calcification.

The application belongs to the technical field of traditional Chinese medicine, and discloses application of matrine in preparation of a medicine for treating endometriosis and a medicine composition. The matrine used in the application can inhibit proliferation and promote apoptosis of ectopic stromal cells of endometriosis, and can also significantly inhibit metastasis and recurrence of the ectopic stromal cells, and can be widely applied in preparation of medicines for treating endometriosis, and has important clinical value.

The present application relates to a kind of drug based on PARM1 regulation treatment calcified aortic valve disease and its application.The present application first finds that PARM1 is significantly increased in the aortic valve of calcified aortic valve disease (CAVD) patient, and plays a key role in the osteogenic differentiation process of valve interstitial cells (VICs), and the expression level of PARM1 is knocked down can inhibit the osteogenic differentiation of VICs and extracellular calcium salt deposition, reduce aortic valve calcification, and its mechanism of action involves MAPK signal pathway.The present application provides a new target and treatment idea for the treatment of CAVD, and can be applied to related drug research and development and treatment method development.

The invention belongs to the technical field of organic synthetic drugs, and particularly relates to a cyclic pyrimidine derivative, a pharmaceutical composition and a preparation method and application of the cyclic pyrimidine derivative. A series of cyclic pyrimidine derivatives are synthesized, and the cyclic pyrimidine derivatives have good sGC agonist activity and can be used as sGC agonists. In-vivo and in-vitro experiments prove that the compound prepared by the invention obviously inhibits osteogenic differentiation of valvular interstitial cells, optimizes in-vivo pharmacokinetic characteristics, obviously reduces aortic valve peak flow velocity and peak pressure difference of a calcified aortic valve mouse, and has a good application prospect. The compound has the effect of relieving arterial valve calcification equivalent to or better than that of positive drugs (Vericiguat and Riciguat) with high administration dosage, and has a good application prospect in the aspect of preparing drugs for treating or relieving aortic valve diseases and other related cardiovascular diseases.

Aortic valve calcification is a condition in which calcium deposits form on the aortic valve in the heart. These deposits can cause narrowing at the opening of the aortic valve. This narrowing can become severe enough to reduce blood flow through the aortic valve—a condition called aortic valve stenosis. The inventors have shown that retinoic acid decreases calcification and osteoblast-like phenotype in valvular interstitial cells (VICs). More particularly, RARα activation reduces calcification and osteoblast-like phenotype in VIC. On the contrary, ALDH1A1 inhibition increases calcification and osteoblast-like phenotype in VIC. Thus the results prompt to consider that use or retinoic acid receptor (RAR) agonists would be suitable for the reversing, preventing or delaying calcification of the aortic valve.

The present invention provides a PAD4 inhibitor, a preparation method thereof, and an application thereof in the preparation of anti-pulmonary fibrosis drugs, belonging to the field of biomedicine technology. The present invention takes PAD4 as a new target for the treatment of pulmonary fibrosis, and performs structural modification on chloramidine to obtain a PAD4 inhibitor having a structure shown in Formula I. The small molecule inhibitor can inhibit PAD4 activity, reduce NETs formation, inhibit the activation of lung fibroblasts and their transformation into myofibroblasts, reduce lung collagen deposition, and slow down the progression of pulmonary fibrosis. In vitro, RC2-33 can reduce histone citrullination to inhibit NETs formation, inhibit TGF-β1-induced lung fibroblast proliferation, and downregulate interstitial marker proteins such as α-SMA, COL-I, FN-I, etc. Nebulized inhalation of RC2-33 can effectively reduce NETs formation in the lungs of mice, inhibit the transformation of epithelial cells to interstitial cells, reduce extracellular matrix deposition, and improve pulmonary fibrosis in mice.

The present invention belongs to the field of biomedicine and specifically relates to a method for inhibiting the osteogenic differentiation of human aortic valve interstitial cells (hVICs). Specifically, the method involves silencing N-acetyltransferase 10 (NAT10) in vitro and in vivo using biological methods. According to this method, NAT10 was over-silenced in mice using an adeno-associated virus, and it was found that NAT10 silencing could inhibit mouse aortic valve calcification. NAT10 was also silenced in hVICs using an adenovirus, and it was found that NAT10 silencing could inhibit the osteogenic differentiation of hVICs, thereby delaying or inhibiting valve calcification. Furthermore, a method for preventing and treating aortic valve calcification by inhibiting NAT10 protein expression is proposed, providing a new non-surgical treatment option for the prevention and treatment of calcific aortic valve disease.

The invention discloses an ovarian endometriosis glandular epithelial organ and interstitial cell co-culture model and a construction method thereof. The construction method comprises the following steps: S1, constructing an ovarian endometriosis cyst glandular epithelial organ; s2, constructing a 3D culture body of the ovarian endometriosis cyst interstitial cells; and S3, digesting the glandular epithelium organ, mixing the glandular epithelium organ with the interstitial cells in proportion, and embedding the glandular epithelium organ in matrigel for three-dimensional co-culture. According to the invention, a patient-derived ovarian endometriosis glandular epithelial organ and interstitial cell co-culture system is established for the first time, the model and a primary focus are highly homologous in tissue morphology and biomarker expression, and the bionic degree is high; it is clear that the number of glandular epithelial organs is the largest when the ratio of epithelial cell clusters to interstitial cells is 1: 2, the vitality of the glandular epithelial organs in a group with the ratio of 1: 2 to 1: 4 is remarkably improved, and the technical bottlenecks that an existing co-culture model is fuzzy in proportion, and the cell viability is difficult to maintain are solved.

This invention relates to an organoid microarray that simulates the immune microenvironment of multiple organs affected by endometriosis. The microarray comprises a chip with three spatially independent cell culture chambers connected by a series channel: an ovarian organoid culture chamber, a peritoneal culture chamber, and an intestinal organoid culture chamber. Each cell culture chamber has a fluid channel in both its upper and lower layers. This invention also provides a method for organ culture using the organoid microarray that simulates the immune microenvironment of multiple organs affected by endometriosis. Its advantages include: simultaneously connecting ectopic endometrial organoids with three target organs; simultaneously simulating the implantation and invasion process of shed endometrial tissue under the influence of the immune microenvironment and in different organs; and constructing a mixed organoid including glandular and stromal cells to explore the interactions and influences between cells of multiple organs and tissues. This is of great significance for preventing endometriosis from affecting vital organs and for discovering new therapeutic targets.

This invention provides a method for isolating and identifying interstitial cells from Bactrian camels, belonging to the field of cell isolation technology. A mixture of 0.08–0.12% collagenase IV solution and 0.23–0.27% trypsin-EDTA at a volume ratio of 1:2 is used to digest Bactrian camel testicular tissue blocks. The mixture is then purified by hypotonic treatment to remove excess germ cells, differential adhesion purification to remove a large number of supporting cells, and Percoll density gradient centrifugation to remove a small number of supporting cells. The suspension, obtained from a 34%–60% Percoll gradient, is transferred to a new culture flask to obtain interstitial cells with a purity ≥95%. Identification is performed using morphological observation, immunofluorescence staining, and secretory function testing. The interstitial cells isolated from Bactrian camels using this invention can be used for research on the reproductive performance of Bactrian camels, laying the foundation for understanding the reproductive performance of Bactrian camels.

The application provides a combined drug for improving embryo implantation efficiency of RIF patients and a screening method, and belongs to the technical field of embryo implantation. In order to solve the problem of how to screen a general drug combination on the basis of a large heterogeneous RIF population, the application locks 1119 kinds of drugs that may have a therapeutic effect in a large range through a six-step screening method combined with advanced RNA sequencing technology, and then further screens the drugs by using a highly simulated endometrial epithelial organoid and stromal cell model, tests the influence of the 1119 kinds of drugs on the activity of the endometrial epithelial organoid and stromal cell, and finally screens 7 kinds of effective drugs which can form a broad-spectrum drug with significant therapeutic effect on RIF in a combined form. The 7 kinds of effective drugs screened by the application are icotinib, fumaric acid propofol tenofovir tablets, clopidogrel, tizacat, osimertinib, venetoclax and rebastinib.