33 results about "Poly d lysine" patented technology

The invention belongs to the technical field of biomedical materials, and particularly relates to a biological adhesive with stable performance and a preparation method and application thereof. Specifically, by regulating and controlling the mixing mode of the active ester and the amino cross-linking agent, an acidic polylysine hydrochloride aqueous solution is adopted as the amino cross-linking agent, the active ester is directly dissolved in the polylysine hydrochloride aqueous solution, and the formed solution mixing system is stable in performance, can be stably stored for 6 hours at room temperature, can be prepared by an operator before an operation, and is free of secondary pollution. And the device can be immediately used at any time in the operation, has good batch stability, and avoids individual difference, so that the device has important application value and market prospect.

The invention discloses a gamma delta T cell efficient amplification method and application, and belongs to the technical field of cell biology. The amplification method comprises the following steps: separating PBMC (peripheral blood mononuclear cells) by adopting a density gradient centrifugation method, carrying out initial culture through a polylysine coated container, carrying out three-stage dynamic stimulation, combining with an X-VIVO 15 culture medium of 5-8% autoserum, and finally carrying out anti-gamma delta TCR magnetic bead separation to obtain high-purity cells. Through collaborative optimization of stepped factor combination, staged container coating and a low-serum system, the amplification multiple of the gamma delta T cells reaches 150-180 times, the purity is larger than or equal to 90% after purification, the cytotoxicity and the survival ability are remarkably improved, the gamma delta T cells can be efficiently used for immunotherapy of tumors and infectious diseases, and stable technical support is provided for clinical transformation of the gamma delta T cells.

The invention relates to the field of biological material science and the field of material science, in particular to a preparation method and application of asymmetric adhesion hydrogel powder for clinical treatment of pleural adhesion. The preparation method comprises the following steps: firstly, preparing hydroformylated hyaluronic acid (HAA) and 5-hydroxydopamine silk fibroin (5-HDP-SF), and preparing lipidosome loaded with simvastatin and miRNA-124 by using a film method; the synthesized HAA, epsilon-polylysine. HCl and 5-HDP-SF are mixed according to a certain mass ratio, sodium hydroxide (NaOH) is added to adjust the pH value, and then adhesive layer hydrogel is formed; dispersing the prepared lipidosome loaded with simvastatin and miRNA-124 into the adhesive layer hydrogel, and then freeze-drying to obtain adhesive layer hydrogel powder (loaded lipidosome); the synthesized HAA and four-arm sulfydryl polyethylene glycol (4-arm-PEG-SH) are mixed according to a certain mass ratio and then freeze-dried, and non-adhesive layer hydrogel powder can be formed; and dissolving the adhesive layer hydrogel powder again to obtain adhesive layer hydrogel, and smearing the non-adhesive layer hydrogel powder on the upper surface of the adhesive layer hydrogel according to a certain mass ratio to obtain the asymmetric adhesive hydrogel (loaded liposome). The hydrogel prepared by the invention is loaded with simvastatin and miRNA-124 at the same time, can be used for synergistically treating pleural adhesion, has good biocompatibility and mechanical adaptability, and has a quite wide application prospect in the field of biological medicines.

The invention relates to the technical field of slide processing. The invention discloses a slide treatment method for improving wall adhesion of 293T cells. The slide processing method comprises the following steps: preparing a cleaning solution; putting the slide into the cleaning solution, soaking, taking out the slide, washing the slide with deionized water, and putting the slide into a drying oven for drying; immersing the dried slide into PLL treatment liquid for 3-7 minutes, taking out and airing; wherein the cleaning solution is prepared from the following raw materials: potassium dichromate, water and concentrated sulfuric acid; the PLL treating fluid is poly-L-lysine with the mass concentration of 0.05%-0.15%, and the solvent is deionized water. The invention further discloses the slide prepared by the slide treatment method for improving the wall adhesion of the 293T cells. According to the slide treatment method, the adhesive force of the slide to 293T cells can be effectively improved, and the cell adherence is enhanced.

The invention relates to the technical field of sustained-release gel preparations, and discloses a sustained-release gel preparation capable of responding to glucose to release growth factors as well as a preparation method and application of the sustained-release gel preparation. According to the invention, a gel preparation containing a'growth factor-FPBA-polylysine-hyaluronic acid 'quaternary composite system is constructed, hyaluronic acid is used as a gel matrix, and FPBA-grafted polylysine and the hyaluronic acid matrix are compounded to form a gel carrier framework; the gel carrier framework is used for realizing mechanical protection and specific condition response type delivery on the growth factors, and the utilization of organisms on the growth factors is realized through a synergistic effect. Wherein the hyaluronic acid can promote the contact between the gel and a wound, the existence of the FPBA-grafted polylysine makes the active component of the growth factor difficult to degrade, and when the gel is in contact with the wound, the glucose responsive release effect of the growth factor is realized through the reversible combination of the FPBA and the glucose; therefore, the gel is suitable for dressing application of diabetes wound healing.

The application discloses a kind of composite drug-loaded nanoparticles based on calcium peroxide carrier, which is formed by the surface of hyaluronic acid modified calcium peroxide nanoparticles loaded with polylysine-dihydrophenophor e6 (Ce6) as carrier. pll The composite drug-loaded nanoparticles CaO2@HA / Ce6 are spherical particles with a particle size of about 100nm. pll In the application, Ce6 is converted into Ce6 by polylysine modification, and then combined with hyaluronic acid modified calcium peroxide nanoparticles. pll Ce6 is uniformly loaded on the surface of the composite drug-loaded nanoparticles CaO2@HA / Ce6. pll The application solves the problem that Ce6 cannot be attached to the surface of CaO2 nanoparticles. pll The prepared nanocomposite particles have uniform particle size and high specific surface area. pll After the combination of Ce6 and CaO2@HA, as a good bacterial microenvironment response material, the anoxic condition of bacterial site can be improved, and the photodynamic therapy can be enhanced. On the other hand, the biocompatibility and delivery efficiency of Ce6 are improved, so that significant bactericidal effect can be achieved at low dose.

The application belongs to the technical field of regenerative medicine, and particularly relates to an ASP / BFP-1 sustained-release nanofiber membrane and a preparation method and application thereof. In the initial stage of bone repair, matrix metalloproteinase-2 (MMP-2) secreted by a large number of stem cells can effectively degrade gelatin and polylysine in the nanofiber membrane coating, so as to sequentially release aspirin and bone formation peptide-1 in the drug-loaded coating. The released aspirin can control inflammation, and the bone formation peptide-1 can enhance the osteogenic potential of stem cells and guide bone tissue regeneration.

The invention discloses an immunofluorescence staining method for karyocytes in blood. When immunofluorescence staining is carried out, polylysine is used for treating a cell slide, and then the cell is fixed and smeared and dried, so that the cell and the slide are attached more tightly. The problem that cells fall off in the cleaning process can be effectively avoided. Meanwhile, non-blood cells in the blood sample can be effectively reserved.

The invention discloses application of desoxyribonucleic acid-poly-L-lysine composite hydrogel in redistribution of endogenous exosomes and treatment of myocardial infarction, and the desoxyribonucleic acid-poly-L-lysine composite hydrogel comprises a DNA (deoxyribonucleic acid) skeleton and poly-L-lysine wrapping the DNA skeleton, the DNA skeleton contains a nucleic acid aptamer targeting exosome surface protein, and the desoxyribonucleic acid-poly-L-lysine composite hydrogel is used for capturing and releasing endogenous exosomes. According to the invention, the injectable desoxyribonucleic acid-poly-L-lysine composite hydrogel is used for treating myocardial infarction, and targeted enrichment and controllable release of endogenous exosomes are realized through the unique molecular design of the desoxyribonucleic acid-poly-L-lysine composite hydrogel; important breakthrough is achieved in the aspects of targeting, controllability, safety, clinical transformation and the like, and a brand new solution is provided for myocardial infarction treatment.

The invention relates to a three-dimensional culture method for hair papilla cells and application of supernate of the hair papilla cells in hair regeneration. The invention relates to a three-dimensional culture method for hair papilla cells. The method comprises the following steps: (1) preparing hair follicle hair papilla cells into a cell suspension; (2) cutting the gelatin sponge, and dropwise adding polylysine; (3) wetting the gelatin sponge with the culture medium; (4) sucking out the culture medium, adding a cell suspension, and incubating, so that the cells are fully colonized; and (5) adding a complete culture medium after colonization, immersing all the sponge, culturing for a period of time, then replacing with a culture medium containing L-ascorbic acid, culturing until cells are aggregated into balls, then replacing with a serum-free culture medium, culturing for 24 hours, and then collecting supernate. According to the method, the in-vivo inductivity of the hair papilla cells subjected to 3D culture by using the gelatin sponge scaffold is maintained, the survival ability and differentiation potential of the cells are improved, and the secretion ability of growth factors for promoting hair regeneration is remarkably improved, so that the effect of promoting hair regeneration is achieved.

The invention belongs to the technical field of medical immunodetection analysis, and discloses a kit for detecting an anti-AQP4 autoantibody and application of the kit. The kit comprises a cell pore plate and a cell freezing medium; reaction holes of the cell pore plate are internally coated with a coating substance, and the coating substance is one of polylysine, gelatin and collagen; the cell cryopreservation solution is a cryopreservation protection solution of cells containing overexpressed AQP4 antigens; during coating, the concentration of a coating substance is 0.5 mg / mL-3. 0 mg / mL, and the dosage of the coating substance is 40 [mu] L / hole to 60 [mu] L / hole. According to the kit for detecting the anti-AQP4 autoantibody, the detection performance can be improved, and the kit can be frozen at low temperature for a long time and then unfrozen for detection.

The invention provides a magnetic nanomaterial-guided anti-inflammatory enhanced mesenchymal stem cell as well as a preparation method and application thereof, and belongs to the technical field of biomedicine. The magnetic nanoparticle labeled mesenchymal stem cell provided by the invention is a magnetic nanoparticle (PLL-MNPs) modified by poly L-lysine coated in a mesenchymal stem cell (MSC). The PLL-MNPs has no obvious influence on the cell morphology, differentiation potential, proliferation, apoptosis and cell cycle of the MSC, meanwhile, the PLL-MNPs obviously enhances the anti-inflammatory ability of the MSC, and the conversion of reactive astrocytes / macrophages from a proinflammatory phenotype to an anti-inflammatory phenotype can be promoted. In conclusion, the mesenchymal stem cells marked by the magnetic nanoparticles have a relatively strong anti-inflammatory function, and a new means is provided for application of clinical cell transplantation therapy of the MSC.

The invention discloses a method for inducing HL-60 cells to generate a neutrophil extracellular trap net and application of the method. The method comprises the following steps: firstly, inducing and differentiating HL-60 cells into neutrophile granulocyte-like cells by using dimethyl sulfoxide (DMSO), and then stimulating the differentiated cells by using phorbol ester (PMA), thereby inducing to form the neutrophile granulocyte extracellular trapping net. Preferably, the final concentration of the DMSO is 0.8%-1.5%, the final concentration of the PMA is 50-400 nM, and the stimulation time is 2-4 hours. According to the invention, a polylysine-coated substrate is adopted to enhance the structural stability of NETs, and SYTOX Green nucleic acid dye is adopted for fluorescence detection. According to the invention, a complete and standardized experimental system formed from the HL-60 cell to the NETs is successfully established, the technical problems of unstable primary cell source and poor experimental repeatability are overcome, the operation is simple and convenient, the result is reliable, and an ideal tool is provided for related research of the NETs.

The application belongs to the technical field of biomedical materials, and particularly relates to a biological adhesive with stable performance and a preparation method and application thereof. Specifically, by regulating a mixing mode of active ester and amino cross-linking agent, using an acidic polylysine hydrochloride aqueous solution as the amino cross-linking agent, directly dissolving the active ester in the polylysine hydrochloride aqueous solution, the solution mixing system formed has stable performance, can be stored at room temperature for 6 hours, an operator can prepare before operation, and can be used at any time during operation, and has good batch stability, avoids individual differences, and therefore has important application value and market prospect.

The application discloses a preparation method of a composite material, which comprises the following steps: S1, obtaining mesoporous silica, adding dopamine, and allowing the dopamine to self-polymerize on the surface of the mesoporous silica; S2, adding a silver source into the reaction system of S1, and carrying out chelation reaction with the polydopamine; S3, adding polylysine into the reaction system of S2; and S4, adding mannose into the reaction system of S3, carrying out coupling reaction with the polydopamine-polylysine, and forming a silver nanoparticle loaded mannose coupled polydopamine-polylysine modified mesoporous silica material. The loading of the polylysine can improve the in-vivo bioavailability and has a broad-spectrum antibacterial performance; the introduction of the mannose coupling can be coupled to the micro-nano material as a target recognition molecule, so that the toxicity generated by off-target is reduced. By utilizing the characteristics that mycobacterium tuberculosis surface highly expresses CD206 receptors, i.e., mannose receptors, and based on the bactericidal effect of silver nanoparticles and Schiff bases, an anti-tuberculosis system of the composite material is constructed.

The invention provides a method for extracting, purifying and identifying fibroblasts, which comprises the following steps of: cutting meninx or spinal membrane tissues into pieces, digesting, and centrifuging to obtain a precipitate; re-suspending the obtained precipitate by using a fibroblast culture solution, spreading the precipitate in a polylysine pre-coated culture flask, culturing until the cell confluence degree is 90%, shaking, and discarding the supernatant liquid to obtain purified fibroblasts; and carrying out passage on the obtained purified fibroblasts, carrying out immobilization and permeabilization, sealing with goat serum, sequentially adding a primary antibody solution and a fluorescent secondary antibody solution, dyeing with DAPI, and observing and identifying the fibroblasts under a fluorescence microscope to complete the extraction, purification and identification of the fibroblasts. The invention also provides an application of the method. The method is used for extracting, purifying and identifying meningeal fibroblasts or spinal fibroblasts. The fibroblasts extracted by the method are high in purity, short in culture period and simple to operate.

The invention provides a compound adjuvant for enhancing cellular immunity as well as a preparation method and application thereof, and belongs to the technical field of biological medicines. The adjuvant disclosed by the invention is prepared from a polylysine hydroxymethyl cellulose polyinosinic cell solution and a PEG (Polyethylene Glycol) 5000-PLA15000-GM-CSF (Granulocyte-Macrophage Colony-Stimulating Factor) nanoparticle solution, the concentration of the polylysine hydroxymethyl cellulose polyinosinic cell solution is 0.25 to 4 mg / mL, and the concentration of the PEG5000-PLA15000-GM-CSF nano particle solution is 0.5 to 2 mg / mL; the volume ratio of the polylysine hydroxymethyl cellulose polyinosinic-polycytidylic acid solution to the PEG5000-PLA15000-GM-CSF nano particle solution is (2.5 to 10): 1. The composite adjuvant disclosed by the invention can activate DC cells and induce stronger immunoreaction, and has a better anti-tumor effect when being applied to tumor treatment. The preparation method of the composite adjuvant is simple and controllable.

The invention discloses a desoxyribonucleic acid-poly-L-lysine composite hydrogel and a preparation method thereof, the desoxyribonucleic acid-poly-L-lysine composite hydrogel comprises a DNA skeleton and poly-L-lysine wrapping the DNA skeleton, and the DNA skeleton contains a nucleic acid aptamer targeting exosome surface protein. By optimizing a cross-linking strategy (such as physical entanglement or chemical covalent bonds), the pore size and the stability of the gel are regulated and controlled, and the DNA aptamer capable of identifying the exosome surface protein in a targeted manner is introduced, so that efficient and specific exosome separation is expected to be realized, the key problems of low efficiency, poor specificity, insufficient stability and the like in exosome separation in the prior art are solved, and the method is suitable for large-scale industrial production. The method has wide application prospects in the fields of liquid biopsy, targeted therapy and the like, and a revolutionary technical platform is provided for clinical application of the exosome.

The invention discloses a female bacteriostatic device, and relates to the technical field of bacteriostatic compositions and bacteriostasis. The device comprises an LED light source, a micro liquid pump and a liquid storage bin, wherein a bacteriostatic composition contained in the liquid storage bin comprises epsilon-polylysine, vitamin B, sodium lignin sulfonate, a zein and hydroxypropyl trimethyl ammonium chloride chitosan compound, calcium peroxide and the like. The composition is prepared into freeze-dried gel through a freeze-drying process, when in use, the freeze-dried gel is redissolved to form a gel state, and the antibacterial effect is enhanced through irradiation of an LED light source. The problem that vitamin B is easy to decompose in the vagina environment is effectively solved, the stability and sterilization efficiency of antibacterial components are improved, and the composition is suitable for prevention and treatment of vulvovaginal candiditis and multi-bacterial mixed vaginitis.

The application discloses a kind of diazirine compound base and preparation method and application thereof, it is related to microarray immobilization compound technical field.The diazirine compound base is obtained by using diazirine compound to modify polylysine base, it is simple in synthesis step, wide in application range, can be applied to high-throughput biochip preparation field, biological molecule interaction research, biological molecule activity evaluation, disease diagnosis, high-throughput screening, drug screening, fluorescence array imaging and surface plasmon resonance imaging etc., with wide market prospect;Meanwhile, when preparing biochip by diazirine compound base, preparation result is more reliable, and the detection result of prepared biochip is more sensitive, with low background and good signal, with universality, and the preparation method of biochip has the characteristics of simple operation, flexible control, without special equipment, low cost, high work efficiency.

The application provides a method for modifying a solid support suitable for adhering cells, comprising: contacting the solid support with a polylysine-protein factor mixture, the polylysine-protein factor mixture comprising polylysine and a protein factor, the protein factor comprising at least one of an extracellular matrix protein and a cell integrin. The method modifies the solid support, allows the solid support to capture a large number of cells, makes the profile of the cells captured by the solid support clear, and improves the cell capture efficiency of the solid support.

The application provides a kind of composite adjuvant for enhancing cellular immunity and its preparation method and application, belong to biological medicine technical field.The adjuvant includes polylysine hydroxymethyl cellulose polymyxin solution and PEG5000-PLA15000-GM-CSF nanoparticle solution;The concentration of the polylysine hydroxymethyl cellulose polymyxin solution is 0.25-4mg / mL, the concentration of the PEG5000-PLA15000-GM-CSF nanoparticle solution is 0.5-2mg / mL;The volume ratio of polylysine hydroxymethyl cellulose polymyxin solution and PEG5000-PLA15000-GM-CSF nanoparticle solution is (2.5-10):1.The composite adjuvant of the application can activate DC cells, induce stronger immune response, and has better antitumor effect when applied to tumor treatment.

This invention discloses a bone cement possessing both antibacterial properties and high mechanical strength. The invention involves the methacrylylation modification of polylysine, followed by a cross-linking reaction to form a hydrogel, which is then mixed with a solidifying agent powder to successfully prepare a novel bone cement. This material not only exhibits excellent antibacterial activity but also possesses superior mechanical properties and good biocompatibility, making it widely applicable in the field of bone tissue repair and regeneration engineering.

This invention discloses a recombinant human (or bovine) basic fibroblast growth factor gel with wound-healing and antibacterial functions, and its preparation method. Recombinant human (or bovine) basic fibroblast growth factor gel is a genetically engineered topical drug used clinically to promote healing of burn wounds, chronic wounds, and fresh wounds, with significant efficacy. However, its drawbacks include limited antibacterial properties and inability to be used simultaneously with chemical disinfectants such as iodine and ethanol, which, while promoting wound healing, are not conducive to controlling wound infection. Research has found that protamine sulfate or polylysine, with high safety and good antibacterial properties, can exert good antibacterial and anti-infection functions without negatively affecting the bioactivity and stability of recombinant human (or bovine) basic fibroblast growth factor. This represents an advanced method for preparing recombinant human (or bovine) basic fibroblast growth factor gel with wound-healing and antibacterial functions, enabling this type of drug to better exert its clinical effects and expand its application scope.

The invention discloses a polymerase labeled mouse and rabbit universal secondary antibody as well as a preparation method and application thereof. The polymerase labeled mouse and rabbit universal secondary antibody comprises an anti-mouse IgG and rabbit IgG recombinant antibody, and the anti-mouse IgG and rabbit IgG recombinant antibody sequentially comprises an anti-mouse IgG nano antibody, a connecting peptide, an anti-rabbit IgG nano antibody, a human IgG hinge region peptide fragment, a polylysine peptide fragment and a purification label from an amino terminal to a carboxyl terminal. According to the present invention, the anti-mouse IgG and anti-rabbit IgG recombinant antibody with the brand new structure is designed to obtain the recombinant antibody with the multiple primary amine groups at the carboxyl terminal, and the primary amine groups can be further utilized to couple the labeled enzyme so as to develop the preparation method of the polymerase-labeled mouse-rabbit universal secondary antibody, such that the polymerase-labeled mouse-rabbit universal secondary antibody can be obtained; the secondary antibody signal amplification effect can be improved, the secondary antibody preparation period is shortened, the secondary antibody preparation process is simplified, the stability between batches is improved, and meanwhile dependence on experimental animals is reduced.

The present invention belongs to the field of medical technology, and in particular to a method for improving the antithrombotic action time and safety of cationic polyphosphate inhibitors in vivo. The present invention specifically chemically modifies cationic polyphosphate inhibitors with human serum protein ligands to improve their antithrombotic action time and safety in vivo, wherein the human serum protein ligand is 2-carboxyl phthalocyanine zinc, and the cationic polyphosphate inhibitor is low molecular weight polyethyleneimine or polylysine. Experimental data show that 2-carboxyl phthalocyanine zinc coupling significantly improves the half-life and anticoagulant action time of low molecular weight polyethyleneimine and polylysine in mice, prevents thrombosis without causing bleeding side effects, and effectively reduces the acute toxicity of low molecular weight polyethyleneimine to mice and zebrafish. Thus, it can be used to develop antithrombotic drugs with broad application prospects.

The invention discloses a light-operated cell membrane chimeric multi-unit drug-loaded liposome and an application thereof. The light-controlled multi-unit drug-loading liposome comprises a cell membrane, a light-sensitive element, a temperature-sensitive lipid material and a multi-unit drug-loading core, the multi-unit drug-loading core comprises a first drug-loading unit, a second drug-loading unit and a third drug-loading unit, wherein the first drug-loading unit is used for entrapping curcumin by taking vitamin E succinate-grafted-polylysine as a material, the second drug-loading unit is used for entrapping trabectedin by taking natural silk fibroin as a material, and the third drug-loading unit is used for compounding doxorubicin hydrochloride by taking dopamine-grafted-gamma-polyglutamic acid as a material. The light-operated cell membrane chimeric multi-unit drug-loaded liposome can reverse the immunosuppressive microenvironment of tumor tissues and improve the drug resistance problem of glioma after multiple chemotherapy, and has a wide application prospect in the aspect of tumor treatment.

The invention provides an establishment method and application of a pseudorasbora parva snout-end cell line, and belongs to the technical field of freshwater fish cell culture.The establishment method comprises the following steps that (1) primary culture is conducted, specifically, pseudorasbora parva snout-end tissue is digested through collagenase II, a cell / tissue suspension is prepared and inoculated into a culture bottle subjected to polylysine coating treatment, and a culture medium is obtained; culturing by using a primary culture medium; (2) subculture: when the cell generations are 1-20 generations, culturing by using a primary culture medium which contains an epidermal growth factor, a chick embryo extract and 20% fetal calf serum; after more than 20 generations, a subculture medium is used for culture, and the subculture medium contains 10% of fetal calf serum and does not contain epidermal growth factors and chick embryo extracts; and (3) collecting passage cells to obtain the pseudorasbora parva snout end cell line. The pseudorasbora parva snout end cell line established by the method can realize continuous passage and proliferation, can be recovered after ultralow temperature cryopreservation, and can be applied to the fields of exogenous gene expression research, water environment pollutant monitoring and the like.

The application discloses a method for identifying extracellular vesicles, and relates to the technical field of identification of extracellular vesicles. The method comprises the following steps in sequence: dropping a polylysine solution on a glass slide, carrying out coating, air-drying, then adding an extracellular vesicle solution, incubating at room temperature, then adding a fluorescein-5-maleimide solution, incubating at room temperature in the dark, and then carrying out fluorescence identification under a confocal microscope. The method realizes rapid detection of the vesicles by using fluorescein-5-maleimide for marking detection.

The invention discloses a method for separating and culturing chick embryo forebrain neurons. The method comprises the following steps: (1) carrying out coating treatment on a cell plate by using a poly-D-lysine solution; (2) obtaining chick embryo brain tissue fluid; and (3) culturing neurons. By improving and optimizing a neuron preparation method and a reagent, the chick embryo forebrain neuron with the high purity of 98% or above is obtained.