97results about How to "Disinhibition" patented technology

The invention discloses a method for producing fatty acid by hydrolyzing lipid through three liquid-phase lipase catalytic systems. The method comprises the following steps: firstly, preparing enzyme solution with the lipase concentration of 5-20000 U/mL, and adding soluble inorganic salts, alcohol/ketonic hydrophilic fraction and lipid to the enzyme solution so as to form three liquid-phase systems; secondly, laying the prepared three liquid-phase systems into an agitated vessel for reaction, standing or centrifuging the systems after the reaction so as to divide the systems into an upper layer, a middle layer and a lower layer, and collecting products in the upper layer so as to obtain fatty acid phase. The method adopts the three liquid-phase systems to extract fatty acid in crude enzyme solution, performs direct hydrolyzing to produce fatty acid, can not only greatly reduce the production cost of enzyme, but also solve the problems of high energy consumption for glycerol and enzyme separation, high cost, high pollution and the like.

The invention relates to a method for preparing PD-1 and LAG3 double knockout CD19 CAR-T cells by Cas9/RNP. The method comprises the following steps: mixing sgRNA and Cas9 protein for knocking out PD-1 and LAG3 genes with an electroporation transformation reagent, and adding the mixture into CD4+/CD8+ cells for electroporation transformation; transfecting CD4+/CD8+ positive cells with CD19 CAR lentivirus; and screening, and culturing and amplifying transfected T cells, so as to obtain the PD-1 and LAG3 double knockout CD19 CAR-T cells. The method, adopting a Cas9/RNP gene editing system, improves the transfection efficiency and the expression efficiency and time of transgenes, simplifies the preparation process of CAR-T cells, and improves the feasibility and increases the load of the system. In addition, the method is free of transfection reagents when the PD-1 and LAG3 double genes are knocked out, and thus the safety is higher.

The invention discloses a gene for encoding anti-BCMA chimeric antigen receptor. The nucleotide sequence of the gene is represented by SEQ ID NO:2. The invention also discloses a recombinant expression vector containing the gene, and a myeloma BCMA antigen-targeted transgenic T cell. The transgenic T-cell is a primitive cell containing the recombinant expression vector and knocked out of a PD1 gene or/and a CTLA4 gene, or is a primitive cell with the chromosome being integrated with the gene for encoding anti-BCMA chimeric antigen receptor and knocked out of tbe PD1 gene or/and the CTLA4 gene.A preparation method of the transgenic T-cell comprises the following steps: mixing of gRNA, CRISPR-cas9 mRNA and HDR mix, and electrotransformation recombination of the T cell. The invention furtherdiscloses an application of the myeloma BCMA antigen-targeted transgenic T cell in the preparation of drugs for treating multiple myeloma. In the construction process of carT of, a recognition sequence of EGFR is introduced, EGFR monoclonal antibody Cetuximab is used to eliminate a carT cell if necessary, and PD1 and CTLA4 genes are knocked out to relieve the inhibition effect of the PD1 and CTLA4 genes on the carT cell and enhance the overcoming effect of the carT cell on the inhibition of the tumor microenvironment on immune cell functions.

The invention relates to a large-scale preparation method of mesenchymal stem cell factors. The large-scale preparation method comprises three steps of invitro culture of mesenchymal stem cells, induction and secretion of the mesenchymal stem cell factors and separation and purification of the mesenchymal stem cell factors, wherein an induction medium required by the step of the induction and secretion of the mesenchymal stem cell factors is prepared from a mixed medium of a DMEM (Dulbecco's Modified Eagle Medium), a PBS buffer solution and an L-alanyl-L-glutamine aqueous solution of which the volume ratio is (30 to 80):(20 to 60):(0.2 to 1.5) and an inducer. According to the large-scale preparation method of the mesenchymal stem cell factors, a single sample source is adopted to culture on a large scale, so that the uniformity of batch products is guaranteed, and the induction medium is adopted in the step of the induction and secretion of the mesenchymal stem cell factors, so that a large quantity of cell factors are stimulated to secrete, a formula is reasonable and effective, the conditions of the whole preparation process are stable and reasonable, and the large-scale preparation method is suitable for large-scale production.

The invention relates to an efficient scale inhibitor of a tail-gas compressor of a styrene device and an application method thereof, comprising hydroxylamine compounds, alcohol amine compounds and phenol compounds. The invention overcomes the disadvantages that the generation of polymer dirt in the tail-gas compressor can not be effectively inhibited by using an ethylbenzene physical spraying method, can thoroughly inhibit and suppress the radical polymerization of the styrene, alpha-methyl styrene, phenyl ethylene and the other activated alkenes in the styrene tail-gas compressor, and has the functions of metallic catalytic coke deposition suppression and detergency and dispersancy. The invention has the advantages of safety and environment protection, energy conservation and low energy consumption and simple use, has no influence on downstream processes, can effectively promote the operational cycle and working load of the styrene tail-gas compressor, reduce energy and material consumption of the styrene device, and can improve the economical and social effects of the whole styrene device.

The invention discloses a method for preparing gardenia blue by utilizing phase-transfer catalysis. The method comprises the following steps: peeling off gardenia jasminoides fruits, crushing into fruit powder, adding ethanol water for supersonic extraction, filtering, performing vacuum concentration on the filtrate, recycling ethanol, drying the solid obtained through concentration, and dissolving the solid into water so as to obtain a crude geniposide product solution; mixing the crude geniposide product solution with beta-glucosidase to be used as a water phase, performing catalytic hydrolysis reaction under the condition of oscillation or stirring so as to obtain an intermediate product genipin by using an organic solvent as an organic phase; adding a solution of hydrophilic amino acid into the organic phase, subsequently reacting under the condition of oscillation or stirring so as to prepare the gardenia blue; concentrating and drying the water phase in vacuum so as to obtain gardenia blue powder. According to the method, a reaction-extraction technique is utilized, product inhibition and side product degradation reaction when the genipin is prepared by using an ordinary method are effectively prevented, and the conversion rate of geniposide is high, and meanwhile high-purity gardenia blue can be obtained through reverse extraction by using an amino acid solution, and an organic solvent can be recycled.

The invention relates to a production process for producing gamma-cyclodextrin by a biological method and belongs to the technical field of cyclodextrin production. The production process solves the technical problems to provide the process for producing gamma-cyclodextrin by the biological method, and the production cost of the gamma-cyclodextrin is reduced. The production process has the technical scheme that the gamma-cyclodextrin glucosyltransferase from alkalophilic bacillus is adopted for producing the gamma-cyclodextrin, the starch pulp regulation is carried out according to the concentration being 10 percent, the materials are stirred for 5 to 15 minutes under the condition being 60 to 90 DEG C, the temperature is set to be 40 to 60 DEG C, the pH is regulated to be about 10.0, the gamma-cyclodextrin glucosyltransferase is added according to a quantity of 1 to 10 units for one gram of starch, after 2 to 5 percent (w/v) organic solvents are added, the sufficient reaction is carried out for 8 to 10 hours, the organic solvents are recovered, and the gamma-cyclodextrin is obtained by adopting a crystallization method.

The invention belongs to the technical field of water environment restoration and particularly relates to a regeneration method for urban sewage by continuous flow. In view of the problems existing in a conventional simultaneous nitrogen and phosphorus removal process of serious shortage of carbon sources due to competition for carbon sources in nitrogen and phosphorus removal, big differences in sludge ages of a denitrifier and phosphorus removal bacteria and demand for dissolved oxygen, insufficient alkalinity of sewage, difficulty in giving consideration to nitrogen and phosphorus removal effects concurrently and the like, through overall planning from the process flow and optimization of tasks of all processing units, the regeneration method can be used for removing phosphorus and organic matters by an anaerobic-aerobic process, partially oxidizing ammonia and nitrogen into nitrite nitrogen by a multistage completely hybrid nitrosation process, and realizing autotrophic denitrification by using an upward-flow anaerobic ammonia oxidation biofilter so as to achieve high-efficient low-carbon treatment and regeneration of the sewage, with effluent COD less than 45 mg/L, BOD less than 10 mg/L, TP less than 0.4 mg/L, and TN less than 10 mg/L, so that the treated sewage can reach the national primary standard A.

The invention discloses a medicine for improving the production performance of a sow and a preparation method and application thereof, and belongs to the technical field of veterinary medicines. The medicine for improving the sow production performance is prepared from 2 to 10 parts by weight of vitamins, 4 to 15 parts by weight of plant extract, 2 to 6 parts by weight of an enzyme preparation, 2to 6 parts by weight of amino acids, 1 to 5 parts by weight of a microbial ecological agent and 1 to 6 parts by weight of mannooligosaccharide. The medicine is easy to use, is effective and safe, hasno residue, effectively reduces the sow production process, promotes the postpartum recovery of the sow, improves ovulation, prolongs the sow production life, promotes the sow mammogenesis and milk secretion, improves milk quality, improves the sow production quality, improves the survival rate of the piglet through use in a lactation period and guarantees the healthy and fast growth of piglets.

The invention relates to a device and a method for biologically manufacturing succinic acid by continuous crystallization, and the device comprises: a bioreactor (1), a cross-flow membrane module (2), and a crystallization separation tank (3); a front end of the cross-flow membrane module (2) is communicated with the bioreactor (1) through a pipeline; a filtering end of the cross-flow membrane module (2) is communicated with a bottom of the crystallization separation tank (3); a top of the crystallization separation tank (3) is communicated with the bioreactor (1); therefore, a loop is formed in the device, and the continuous fermentation and crystallization of succinic acid is realized; a peristaltic pump (9) is disposed between the bioreactor (1) and the cross-flow membrane module (2); a back end of the cross-flow membrane module (2) is communicated with the bioreactor (1), which realizes the circulation of the flowing fermentation liquid between the two. The device and the method for biologically manufacturing succinic acid by continuous crystallization of the invention establish a coupling system for low-temperature crystallization in-situ extraction of succinic acid and fermentation and separation, and meet requirements for disinfection sterilization and pollution-free operation.

The invention discloses a method for expressing and purifying recombinant ethanol oxidase rAOX. According to the method, an ethanol oxidase-poly-L-histidine expression vector pPIC9K-AOX-HIS is constructed first by a molecular biological technique, then the expression vector pPIC9K-AOX-HIS is linearized and transferred to pichia pastoris GS115, and thus, pichia pastoris engineering bacteria capable of expressing active recombinant ethanol oxidase are obtained, and part of recombinant ethanol oxidase can be secreted and expressed. Through optimization of the expression conditions of recombinant ethanol oxidase, the ethanol oxidase produced by the engineering bacteria may reach 1,862U/L. Meanwhile, the secretion and expression of the ethanol oxidase and the introduction of a poly-L-histidine tag simplify the purification process of the recombinant ethanol oxidase. According to the result of tests, the recombinant ethanol oxidase expressed by the engineering bacteria has much higher thermostability than pichia pastoris wide type ethanol oxidase. The operation in the whole process of the method is simple, so the method is suitable for industrial production and can create certain economic benefit.

The invention discloses a dry anaerobic fermentation process for reducing ammonia nitrogen accumulation. The dry anaerobic fermentation process is characterized by comprising the following steps of: detecting ammonia nitrogen concentration of fermentation liquor in a dry anaerobic fermentation reaction tank; when the ammonia nitrogen concentration of the fermentation liquor is less than a preset value, directly reflowing into the dry anaerobic fermentation reaction tank; and when the ammonia nitrogen concentration of the fermentation liquor reaches the preset value, removing ammonia nitrogen of the fermentation liquor through zeolite. The invention also discloses a device which is used for implementing the process. According to the dry anaerobic fermentation process and device, the suppression of the high-concentration ammonia nitrogen on the anaerobic fermentation reaction is reduced, the gas producing speed of the system is increased, and the replaced zeolite which adsorbs the high-concentration ammonia nitrogen can also be used as a soil conditioner.

The invention discloses a method for producing lactic acid and simultaneously realizing coupling preparation of GABA (gamma-aminobutyric acid). The method comprises the steps of inoculating rhizopus oryzae into a fermentation culture medium to perform fermentation, adding glutamic acid decarboxylase, pyridoxal phosphate and L-sodium glutamate into a fermentation solution when the pH value of the fermentation solution is less than 4.8, then continuously adding L-sodium glutamate into the fermentation solution, and generating GABA of glutamic acid decarboxylase by utilizing the vitality of catalyzing glutamic acid to consume H+ in an environment so as to maintain the pH value of the fermentation solution at 4.8-6.0 and achieve the purposes of relieving acid inhibition against fermentation of lactic acid and realizing simultaneous production of lactic acid and GABA. The invention provides a new method for relieving the inhibition of a substrate for fermentation of lactic acid; furthermore, in the fermentation process, the coupling production of a functional compound, namely gamma-aminobutyric acid, is realized, so that the yield of lactic acid is increased, and the economy of the process is further improved.

The invention discloses a medicinal Chinese medicine for curing regenerative obstructive anaemia and its preparing technique, prepared in proportion of red ginseng tails, Radix astragali, Radix rehmanniae preparata, dodder seed, alter slice, moxibustion tortoise plastron, medlar, Malaytea Scurfpea Fruit, Prepared Common Monkshood Daughter Root, and cinnamon. And the preparing technique: decocting the above ten raw materials twice and combining the two filtrates; then concentrating, alcohol-depositing, cooling to room temperature and making into cream; drying the cream and then making powder spraying and filling into capsules; or adding dextrin or starch to the cream to make into soft material, and finally screening and drying the soft material to make into electuary or tablets. It can not only regulate mediation disorder of human immunity, stimulate the hemopoiesis recovery of human marrow, and have remarkable curative effect on the regenerative obstructive anaemia but also has considerably good curative effect on other refractory anaemias, as well as the symptoms caused by various reasons, such as marrow inhibition, blood cell reduction, etc. And it almost has no toxicity and side effect, safe and reliable, low-cost.

The invention discloses a comprehensive extraction method for fructus gardenia. The comprehensive extraction method comprises the following steps: carrying out ultrasonic extraction on the fructus gardenia by taking water as a solvent so as to obtain supernate and solid residues, filtering the supernate by virtue of a micro-filtration membrane to obtain filtrate A containing geniposide and a gardenia yellow pigment, mixing the filtrate A with beta-glucosidase liquid as a water phase, converting geniposide in the filtrate A into genipin by virtue of beta-glucosidase in a two-phase system, and transferring the genipin into the organic phase, namely an organic solvent, so as to react to obtain a gardenia blue pigment; separating and purifying the gardenia yellow pigment in the residual water solution; and extracting the solid residues with ethanol to obtain supernate, and carrying out separation and purification, so as to obtain ursolic acid. The comprehensive extraction method has the beneficial effects that the cost is low, the process is simple and practical, efficient paths for purifying the gardenia yellow pigment and converting the geniposide into the gardenia blue pigment are realized, and the extraction of ursolic acid in the solid residues, the recycling and repeated use of fixed beta-glucosaccharase and the maximum utilization of the fructus gardenia are realized.

The invention discloses a preparation method of tumor-enhanced tumor-infiltrating lymphocytes. The preparation method comprises the following steps: (1) extracting tumor-infiltrating lymphocytes TILs from tumor tissues; (2) sorting and enriching PD1 positive T cells in the TILs; (3) integrating a PBR transformed fusion protein and an IL-15 super compound into the PD1 positive T cell at one time; and (4) carrying out multiplication culture to obtain the tumor enhanced TILs. The preparation method has the advantages that (1) by sorting the PD1 positive T cells, the proportion of tumor-specific T cells is increased; (2) the designed PBR fusion protein can relieve the inhibition from a PD1 pathway in a tumor microenvironment and improve the killing ability of the TILs to tumor cells; (3) the IL-15 super compound not only can improve the tumor killing activity of T cells, but also can stimulate other immune cells in the body, such as endogenous NK, T cells and the like, and accelerates the removal of the tumor cells in the body; and (4) the added 'suicide gene' R structure solves the problem of subsequent clinical safety after T cell gene modification.

The invention discloses a composition for removing tumor immunosuppression and application thereof. The composition provided by the invention comprises following (a) to (e) components or coded nucleicacid thereof: (a) IL-15, a mutant thereof or functional fragments thereof; (b) an IL-15 receptor alpha, a mutant thereof or functional fragments thereof; (c) IL-12, a mutant thereof or functional fragments thereof; (d) TGF-beta regulatory peptide, a mutant thereof or functional fragments thereof; and (e) PD1-CD80 fusion protein, a mutant thereof or functional fragments thereof. According to the composition provided by the invention, the immune response of T lymphocytes can be improved synergistically.