30 results about "Cellular formation" patented technology

The invention belongs to the technical field of biological medicines, and particularly relates to application of a macrophage Angulin-1 gene and / or an Angulin-1 protein in preparation of medicines and diagnostic products for preventing and / or treating atherosclerosis and related diseases. Research finds that plaque and necrotic core areas of an atherosclerosis animal model are remarkably increased due to macrophage Angulin-1 gene knockout, and disease development is promoted; and the recovery of the expression level of Angulin-1 significantly inhibits the development of lesion. Cell experiments show that Angulin-1 reduces intake of oxidized low-density lipoprotein by down-regulating expression of macrophage LOX-1, so that formation of foam cells is inhibited. Therefore, the macrophage Angulin-1 can be used as an atherosclerosis drug target, a gene therapy target gene and an auxiliary diagnosis marker, and a new theoretical basis and an intervention strategy are provided for prevention and treatment of the disease.

The present: invention relates to a pharmaceutical composition comprising curcumin and ginsenoside, and a formulation comprising same. Using curcumin and ginsenoids according to the present invention in combination with butyric acid and sucralfate restores cell growth, mobility, and endothelial cell tube-forming by ability reduced irradiation, and inhibits an inflammatory response induced in macrophages by LPS treatment. In addition, it was confirmed that, when an irradiated mouse is topically treated with the formulation according to the present invention comprising the compounds, intestinal epithelial tissue damaged by irradiation is restored and the expression of inflammatory cytokines in the intestinal tissue is suppressed. Furthermore, a mixture of a ginseng extract and a turmeric extract also restored the intestinal epithelial tissue damaged by irradiation. Therefore, the pharmaceutical composition comprising curcumin, ginsenoid, butyric acid, and sucralfate, or the pharmaceutical composition comprising a mixture of a ginseng extract and a turmeric extract, and the formulation according to the present invention comprising same can be utilized for treating radiation proctitis.

The present disclosure relates to a method of generating pancreatic lineage cells, e.g. islet-like cell aggregates comprising pancreatic beta cells, comprising the steps of providing a single cell suspension of a population of endocrine progenitor cells (EP), allowing the EP cells in the single cell suspension to form 3D structures, and culturing the cells under conditions allowing differentiation into pancreatic mono-hormonal beta cells. The present disclosure also relates to islet-like cell aggregates obtainable by said method and their medical use.

The present application relates to a method for preparing skeletal muscle organoids, comprising the steps of: inducing paraxial mesoderm stem cells (PMSCs) by directly differentiating human pluripotent stem cells (hPSCs); forming the paraxial mesoderm stem cells in a 3D shape, and then culturing same in a medium; inducing the proliferation of the paraxial mesoderm stem cells; and inducing the maturation of the paraxial mesoderm stem cells.

B cell proliferation and / or memory B cell formation / expansion can be enhanced by contacting MAIT cells with nogapendekin alfa invaxcept (N-803) to produce stimulated MAIT cells, which stimulate B cell proliferation and / or memory B cell formation / expansion. Such stimulation can be performed in the presence of antigen or antigen-presenting cells. In particularly contemplated embodiments, MAIT cell stimulation is performed in vitro to produce immunostimulatory compositions and vaccines, or in vivo to enhance immune responses in airway tissue. Most typically, in vivo immune stimulation is performed by inhalation or intranasal delivery of a composition comprising N-803 and, optionally, vaccine components.

The invention relates to an establishment method of a high-fat induced intestinal organ injury model and an injury intervention method for the model. The establishment method comprises the following steps: step A, preparation: preparing colon cells, PBMC cells and an organ-like culture chip for later use; step B, colon barrier construction: inoculating colon cells to the surface of a barrier membrane on the lower side of a culture chip by using an organoid culture chip to obtain a colon barrier model; step C, constructing a co-culture model, and inoculating PBMC cells to the basal side of the colon barrier on the basis of the colon barrier to obtain a human colon barrier-PBMC co-culture model; d, constructing a damage model, and performing damage treatment on the co-culture model by using a treating fluid to obtain the damage model; according to the invention, the barrier function organ-like chip is used, a human intestinal cell-PBMC co-culture model is formed by utilizing human intestinal organs and PBMC cells, and OXLDL / palmitic acid can play a role in an injured intestinal organ barrier model and form a high-fat induced intestinal organ injury model through the prepared injury treating fluid.

The invention discloses a bionic gastric mucosa micro-barrier structure, comprising: a bottom microfluidic channel for simulating a submucosal vascular network; the middle gastric epithelial cell layer is used for simulating a mucous membrane epithelial barrier and comprises a matrix layer, quaternary functional cells, a culture medium and a pH buffer module; and the surface dynamic mucus layer is used for simulating a mucous membrane protective barrier and comprises a vagus neurotransmitter simulant, exogenous mucus and a pressure sensor. According to the invention, continuous nutrition supply is provided for the middle gastric epithelial cell layer through the bottom microfluidic channel, the survival and functions of cells are maintained, and accurate tracking of the transgastric mucosa transport process is realized. A function collaborative network is formed through quaternary cells, the secretion volume and the injury state are monitored in real time, regulatory factors are dynamically adjusted, and it is ensured that a cell layer can still maintain the barrier function stable under external interference. When foreign matter impact or food friction signals are recognized through the pressure sensor, the pump speed is instantly increased, and local mucus loss is rapidly supplemented.

The application discloses application of FTY720 as a PP2A phosphatase activator in preparation of a medicine for treating type I neurofibromatosis. The application first uses FTY720 for treatment of type I neurofibromatosis, and proves that FTY720 significantly inhibits formation of neurofibromas by non-specifically activating PP2A phosphatase. In-vivo experimental results show that FTY720 as a single drug can significantly inhibit tumor growth, and a synergistic effect is presented when FTY720 is combined with a MEK inhibitor, and tumor growth is almost completely inhibited. In-vitro cell experiments show that FTY720 as a single drug or in combination with MEKi treatment can inhibit tumor Schwann cells from forming tumor spheres, inhibit cell proliferation and migration, and induce tumor cell apoptosis. The application overcomes the drug resistance problem existing in the prior art MEK inhibitor, and provides a new treatment strategy for type I neurofibromatosis. FTY720 is an FDA-approved drug, and has good safety and drugability, and has high clinical conversion potential.

In some embodiments, B cell proliferation and / or memory B cell formation / proliferation may be enhanced by contacting MAIT cells with Noogatointerleukin alpha due to chibercept (N-803) to produce stimulated MAIT cells, which in turn stimulate B cell proliferation and / or memory B cell formation / proliferation. Such stimulation may be performed in the presence of an antigen or antigen presenting cell. In particularly contemplated embodiments, MAIT cell stimulation is performed in vitro to produce an immunostimulatory composition and vaccine, or in vivo to enhance an immune response in airway tissue. Most typically, in vivo immune stimulation is carried out by inhalation or intranasal delivery of a composition comprising N-803 and optionally a vaccine component.

[Object] Provided is a culture solution for promoting immune improvement and normal immune formation that promote healthy immune formation of humans and cure allergic diseases such as atopic dermatitis, and a culture solution generator capable of easily generating the culture solution. [Solving Means] A culture solution generator 1 generating a culture solution for promoting immune improvement and normal immune formation, the culture solution obtained by suspending 300 g or more of Bacillus fermented powder in 150 liters of water, maintaining a temperature of 30 to 45°C, and supplying oxygen with an oxygen saturation of 70% or more for fermentation.

The invention relates to the technical field of androgen-derived alopecia, and discloses a micro-needle patch based on a hierarchical targeting mechanism as well as a preparation method and application of the micro-needle patch. The finasteride is arranged on the needle tip section of the microneedle, the quercetin is arranged on the needle root section, different cells are targeted through the finasteride and the quercetin in a graded mode, and the microneedle patch with a graded targeting mechanism is formed. After the micro needle pierces into the skin, the water-soluble polymer at the needle tip section is dissolved, finasteride targeted hair papilla cells are released, and then in the process that finasteride acts, the quercetin slowly released at the needle root section gradually acts on endothelial cells in a targeted mode, the proliferation and migration activity of the endothelial cells is promoted, and the effect of the finasteride on the skin is achieved. Endothelial cells are enhanced to form a highly branched capillary network to act on hair papilla cells, the metabolic demand of the hair papilla cells is maintained, and finally the effect of enhancing the finasteride targeting effect is achieved. Therefore, the microneedle patch with the hierarchical targeting mechanism has an excellent AGA curative effect.

The present disclosure relates to a method for the generation of cells of the pancreatic lineage, for example pancreatic islet-like cell aggregates comprising pancreatic β-cells, which method comprises the steps of providing a single cell suspension of a population of endocrine progenitor (EP) cells, allowing said EP cells in single cell suspension to form 3D structures and culturing said cells under conditions permissive of differentiation into pancreatic monohormonal β-cells. The present disclosure also relates to pancreatic islet-like cell aggregates obtainable by said method as well as to medical uses thereof.

The invention relates to a covalent nucleic acid aptamer. The covalent nucleic acid aptamer comprises any one of S2 nucleic acid aptamer DNA sequence as shown in SEQ ID NO: 1, S2 nucleic acid aptamer DNA sequence as shown in SEQ ID NO: 2; the DNA sequence of the S2v1 aptamer is as shown in SEQ ID NO: 2; and the DNA sequence of the S2v2 aptamer is as shown in SEQ ID NO: 3. The aptamer also comprises phosphorothioate modification at the 3'end of the S2v2 and addition of an electrophilic group. In the aptamer, six T basic groups are added at the 3'end of S2v2, and phosphorothioate modification is carried out. Further, the aptamer reacts with 4-(bromomethyl) benzenesulfonyl fluoride to modify an electrophilic group, and the aptamer can form covalent binding with cells. The nucleic acid aptamer has high specificity, high affinity and no immunogenicity, can be chemically synthesized in vitro, can target bladder cancer cells, and is relatively strong in binding capacity. In addition, the aptamer can kill or inhibit bladder cancer cells, the effect is obvious, and targeted therapy of bladder cancer is facilitated.

The invention belongs to the technical field of primary cell culture, and particularly relates to a bronchial epithelial cell physiological or pathological model and a construction method and application thereof.The construction method comprises the steps that primary HBECs are provided, an amplification culture medium is used for conducting infiltration type amplification culture on the HBECs in a cell culture plug-in, after the cells form a complete barrier, the amplification culture medium is removed, and the model is obtained; carrying out gas-liquid interface culture on the cells in the cell culture plug-in by adopting a differential culture medium so as to induce the differentiation of the HBECs to form a false multi-layer columnar epithelial tissue structure; the cell culture plug-in comprises a porous membrane substrate, and construction of a physiological model or a pathological model is realized by selecting the cell culture plug-in of the porous membrane substrate with different pore diameters. According to the method, the in-vitro differentiation period of the HBECs can be remarkably shortened, and the model culture efficiency is greatly improved. The bronchial epithelium model capable of accelerating physiological differentiation or having pathological phenotype prepared by the method provided by the invention has significant advantages in drug screening and disease research.

The invention relates to the field of biological medicines, in particular to a TCR55 mutant and application thereof.The TCR55 mutant comprises an alpha chain and a beta chain, the TCR55 mutant is obtained after mutation is conducted on the basis of wild TCR55, and the alpha chain of the TCR55 mutant is a mutant obtained after any one of the following sites on the alpha chain of the wild TCR55 is mutated into histidine, the fourth position D and / or the fifth position E of the CDR2 alpha, and the seventh position A of the CDR3 alpha; or the beta chain of the TCR55 mutant is a mutant obtained by mutating any one of the following on the beta chain of the wild type TCR55 into histidine: the second site N of CDR1beta, the second site A of CDR2beta, the fourth site E and / or the sixth site T of CDR2beta, and the thirteenth site Y of CDR3beta. The mutant can be used for identifying HIV (Pol448-456) / HLA-B35, has high activating power, triggers downstream signal transduction, and can be used for transforming T cells to form specific high-activating-power TCR-T cells for T cell adoptive immunotherapy.

The application relates to the fields of biotechnology and targeted therapy, and particularly relates to a recombinant construct, a CAR-T cell and construction and application thereof. The application verifies the role and mechanism of THEMIS in maintaining the long-term killing of tumor cells of CAR-T through experiments. The CAR-T cell overexpressing THEMIS is constructed, compared with a control group, the proliferation capacity and memory cell formation capacity of the THEMIS-7x19 CAR-T cell are enhanced after antigen stimulation, and the cytokines are in a controllable state, the cell apoptosis and exhaustion are reduced, and the long-term tumor killing capacity can be maintained more effectively. The in-vivo experiment also proves that the THEMIS-7x19 CAR-T cell has stronger anti-tumor effect compared with the control group in a lymphoma tumor-bearing mouse model, and the survival time of the mouse is significantly prolonged.

The invention provides an induced differentiation method of a pluripotent stem cell derived kidney organoid, which is realized through five key stages: firstly, inoculating pluripotent stem cells into a 24-pore plate coated with matrigel, so that the cells form a single cell adherent state with proper density; a WNT agonist and a Noggin factor are added in the Day 0-4 stage to induce original stripe differentiation; the cells are inoculated into the low-adhesiveness U-shaped bottom plate in the Day4 stage to form spheres; in the Day 4-7 stage, FGF9 and Activin A factors are added to induce an intermediary mesoderm; at the Day 7-11 stage, adding FGF9 to induce kidney mesenchymal; and factors do not need to be added to promote the formation of renal units in the Day 11-18 stages. According to the method, the whole differentiation period does not exceed 20 days, matrigel embedding is not needed in the whole process, the organoid capable of simultaneously expressing LRP2, PODXL and E-cadherin can be obtained, and the organoid contains glomerular-like clusters and renal tubule-like structures. The method has the characteristics of simplicity and convenience in operation, short period and low cost, and can be used for kidney development research, drug screening and disease model construction.

The application discloses a HeLa cell three-dimensional culture model, a preparation method and application. HeLa cells are mixed with a collagen-sodium alginate composite hydrogel precursor solution to obtain a cell-precursor mixture, the cell-precursor mixture is mixed with a CaCl2 solution to obtain a mixed solution, and the mixed solution is placed in an incubator for standing culture for 10-60 min, so that the hydrogel is cross-linked and solidified to form a three-dimensional culture system; complete culture medium is further added for continuous culture, and a HeLa cell three-dimensional culture model is constructed. The HeLa cell three-dimensional culture model is constructed by using the hydrogel, defects existing in simulation of a tumor microenvironment by two-dimensional culture and an animal model are effectively solved, the body microenvironment can be better simulated, cells form regular three-dimensional cell spheroids, a malignant phenotype is maintained, the accuracy and repeatability of drug screening are improved, and a reliable technical platform is provided for cervical cancer mechanism research and anti-tumor drug screening.

The invention provides an application of nevermectin in tumor treatment. According to the present invention, the glucosidase II catalytic subunit GANAB can be subjected to targeted inhibition with the affinity constant KD of 0.312 [mu] M, the effect of the 3-site methoxyl demethylation product Norhistidine of the nehistidine on the GANAB is weak, and the affinity constant KD of the Norhistidine is 97.59 [mu] M; neurotetrastin can inhibit GANAB to induce endoplasmic reticulum stress and unfolded protein reaction of tumor stem cells, promote differentiation of the tumor stem cells, induce death of the tumor stem cells and play a role in inhibiting malignant tumors. In a glioblastoma stem cell model, the cytotoxic activity IC50 of the nevermectin is 7.20 + / -2.10 [mu] M, and a mouse in-situ brain tumor formed by glioblastoma stem cells can be significantly reduced after intratumoral injection, indicating that the nevermectin has good anti-tumor stem cell activity.

The invention relates to the field of biotechnology and targeted therapy, in particular to a recombinant construct, a CAR-T cell and construction and application of the CAR-T cell. Experiments verify that THEMIS maintains the effect and mechanism of CAR-T for persistently killing tumor cells. The CAR-T cell overexpressing THEMIS is constructed, compared with a control group, the multiplication capacity and the memory cell forming capacity of the THEMIS-7 * 19 CAR-T cell after antigen stimulation are enhanced, cell factors are in a controllable state, cell apoptosis and depletion are reduced, and the more effective long-term tumor killing capacity can be maintained. In-vivo experiments also prove that the THEMIS-7 * 19CAR-T cell has a stronger anti-tumor effect in a lymphoma-bearing mouse model compared with a control group, and the survival time of the mouse is remarkably prolonged.

The invention relates to cell biology and regenerative medicine, specifically to a culture method and application for porcine limbal epithelial stem cells. The method involves pretreatment of cells, coating of culture dishes, centrifugation, culturing, passaging, and identification by immunofluorescence staining. The cultured cells are applicable in repairing corneal diseases, including limbal stem cell deficiency. During repair, the cells can be combined with biological materials to form a stem cell engineering carrier for implantation at the injury site. The separation and culture method is simple, yields a large number of cells, and is suitable for use with porcine eyeballs. Limbal epithelial stem cells cultured in vitro form distinct cell clusters, referred to as stem cell islands. The obtained cells exhibit phenotypes including BMI1+, ΔNp63α+, CK14+, and ABCG2+. Based on experimental and clinical research, this method provides a promising approach for cell transplantation in ocular surface reconstruction.

Disclosed is a method for producing kidney organoids including steps of: (1) differentiating stem cells into metanephric mesenchyme cells; (2) forming metanephric mesenchyme cell aggregates by culturing the metanephric mesenchyme cells; and (3) differentiating the metanephric mesenchyme cell aggregates into kidney organoids.

The invention relates to a ZIF-8 nano-enzyme with a Ce-luteolin coordination coating as well as a preparation method and application of the ZIF-8 nano-enzyme, and belongs to the technical field of biological medicines and nano-materials. The nano enzyme is constructed by adopting a step-by-step loading-coordination assembly strategy: firstly, synthesizing ZIF-8 nano particles as an inner core; then luteolin is introduced and adsorbed to pores and the surface of ZIF-8; finally, cerium ions are introduced, and the dense Ce-luteolin metal-organic coordination coating is formed on the surface of the ZIF-8 through in-situ growth by means of the strong coordination effect of phenolic hydroxyl groups and carbonyl groups in luteolin molecules and the cerium ions. The nano-enzyme has excellent SOD-like and CAT-like activities, and effectively inhibits foam cell formation, regulates endothelial cell senescence-related secretory phenotype (SASP) and alleviates DNA damage through an ROS removal-anti-inflammatory-anti-aging synergistic mechanism. The invention provides a new multi-target nano intervention strategy for the treatment of atherosclerosis.

The present application relates to the technical field of tissue engineering, and particularly relates to an organoid double-layer three-dimensional tissue graft for treating retinal pigment degeneration and a preparation method thereof.The organoid tissue graft of the technical scheme comprises a retinal precursor stem cell layer, a retinal pigment epithelial cell monolayer cell layer and an electrospun silk support layer arranged in sequence.The retinal precursor stem cell layer is formed by solidification of a low-temperature agarose cell suspension containing retinal precursor stem cells.The organoid tissue graft of the scheme can supplement a plurality of degenerative missing cells, can form interaction between transplanted cells and can maintain the activity of the transplanted cells.The three-dimensional retinal stereoscopic structure formed can guarantee correct nerve circuit and improve the visual function of the patient.The electrospun silk material provided by the scheme forms support for cells, and prevents deformation of the graft in the later period.The technical scheme can solve the technical problem that the treatment effect of the organoid tissue graft of the prior art on retinal pigment degeneration is not ideal, and has an ideal popularization and application prospect.

The invention relates to the technical field related to biological medicine, in particular to a method for inhibiting cholesterol esterification of macrophages, through targeted regulation of a PPAR-gamma / LXR-alpha signal channel by means of palmatine (PAL), foam cell formation can be effectively reduced, and accordingly atherosclerosis (AS) can be improved. Oxidized low-density lipoprotein (ox-LDL) is used for inducing foaming, then 25-100 [mu] g / ml palmatine is used for intervening for 48 hours, and experiments show that the palmatine remarkably reduces the cholesterol esterification rate (from 52.17% to 41.88%, Plt; according to the present invention, with the application of the palmatine in the preparation of the medicine, the expression of cholesterol efflux genes such as ABCA1, ABCG1 and the like is up-regulated, the release of inflammatory factors IL-6 and TNF-alpha is inhibited (reduced by more than 60%), and the molecular docking proves that the combination of the palmatine and the PPAR-V is stable (the combination energy is-8.482 kcal / mol), and is superior to the traditional medicine; the invention provides a new strategy for natural, safe and multi-target intervention of AS, and has significant clinical application potential.

The invention relates to a dibenzocyclooctyne modified tumor antigen, an anti-tumor vaccine and application of the dibenzocyclooctyne modified tumor antigen and the anti-tumor vaccine. The DBCO modified tumor antigen has an adjuvant effect, and the DBCO modified tumor antigen or the protein vaccine can promote DC antigen presentation and T cell activation, improve CTL killing activity and promote memory T cell formation by activating an NF-kB / iNOS signal axis, so that the tumor immunoprophylaxis and treatment effects of the tumor vaccine can be remarkably improved. Meanwhile, the DBCO modified tumor antigen or the tumor vaccine can remodel the tumor microenvironment through lung tissue targeting, significantly inhibit the progress of pulmonary metastatic tumors, and can synergistically enhance the curative effect of treating tumors by being combined with an immune checkpoint inhibitor. The invention provides a novel immune regulation strategy for the development of chemically modified vaccines, and has important significance in the field of anti-tumor treatment.

The invention relates to methods of preparing an implant for bone repair, the method comprising the steps of expanding periosteum derived cells, mixing the expanded periosteum derived cells with a partially cross-linked sodium alginate solution, forming the alginate solution and the cells into a shaped construct, differentiating the cells in the shaped construct towards chondrogenic differentiation, and dissolving crosslinked alginate with a calcium chelating agent.

The application belongs to the technical field of biomedical material engineering, and discloses a hydrogel support for 3D tumor sphere culture and drug screening, which is formed by four-arm polyethylene glycol maleimide, fibrinogen and thrombin through Michael addition cross-linking and biological polymerization cooperation, and has an adjustable elastic modulus of 10-1000 Pa by adjusting the proportion of each component, and the gelation is completed within 5 minutes at room temperature. The support has the biological activity of fibrin and the structural stability of polyethylene glycol, can efficiently support HCT116 cells to form high-vigor tumor spheres, accurately simulates the in-vivo tumor microenvironment, significantly improves the accuracy and repeatability of drug screening, is simple to operate and compatible with the high-throughput platform of standard well plates, and is suitable for the drug sensitivity detection of tumor personalized treatment.

The invention discloses a bispecific antibody DLK1 / CD3 for treating liver cancer. The bispecific antibody DLK1 / CD3 comprises an anti-human DLK1 monoclonal antibody and an anti-human CD3 monoclonal antibody. The sequence of the single-chain variable region of the DLK1 monoclonal antibody and the single-chain variable region of the CD3 monoclonal antibody is VL (DLK1)-VH (DLK1)-VH (CD3)-VL (CD3). The bispecific antibody can form immune synapses by targeting tumor cells and T cells, directly activate the T cells and enhance the killing ability of the T cells to liver cancer cells.

The invention discloses a PGLYRP2 cytoplasm isomer and application thereof, relates to the field of biological medicines, and aims to solve the problem that drug resistance and immune escape are easy to generate when an existing drug is used for treating hepatocellular carcinoma. The cytoplasm isomer is PGLYRP2 [beta], compared with PGLYRP2, the cytoplasm isomer has the advantages that a nuclear localization signal sequence is deleted, a disordered structural domain IDR1 and a complete PGRP structural domain are reserved, and an inherent disordered structural domain IDR2 is added to the C end of the PGLYRP2 [beta]. Membrane-free condensation bodies can be formed in cytoplasm through liquid-liquid phase separation, and brand new functions different from those of nuclear type PGLYRP2 are given to the membrane-free condensation bodies. The PGLYRP2 beta induces ferroptosis and activates antigen presenting cells and CD8 + T cells, so that a self-amplification positive feedback mechanism of ferroptosis-immune activation is formed. The tumor growth is obviously inhibited; the PD-1 immunotherapy drug resistance is reversed. The PGLYRP2beta has a definite prognosis prediction value, and shows the potential application of the PGLYRP2beta in precision medicine.