61results about How to "Strong secretory ability" patented technology

The invention provides a sludge composting method, and relates to a sludge treatment method which comprises the steps of preferable accessory selection, CK21 fungus spraying inoculation, heavy metal passivation and powder granulation, and then three-dimensional composite biological organic fertilizer with organic, inorganic and microbial active constituents is obtained. Mushroom compost and pig manure are used as accessories which are easily obtained and degraded, the stacker fermentation time is short, only one time of fermentation is needed, the C/N ratio can be effectively adjusted, and the water content of the sludge is reduced; and the efficient beneficial bio-fungus community CK21 is adopted. The microbial concentration is high, the secretion ability of the enzyme is strong, organic matters can be quickly decomposed so as to prevent the corruption of the organic matters, no odor exists, sludge is used for CK21 inoculation, fermentation starts fast, and the effect is good; and the product is inoculated with the CK21 fungus secondarily so as to further balance and activate nutrients, and the microbial natural reproduction is utilized to realize the is called flower dipping absorbing of the plants to nutrients. The invention has the advantages of optimizing the composting conditions, shortening the composting time, reducing energy consumption of turning, and reducing the damage of the heavy metal on the ecological environment.

The invention relates to the technical field of cell culture, and particularly relates to a culture method of mesenchymal stem cells. In the culture method, a serum-free culture medium optimized by the invention is used for culturing the mesenchymal stem cells. In addition, the invention further provides a better culture method for 3D culture. The cells obtained by the culture method disclosed bythe invention are strong in proliferation capacity, intact in shape and strong in secretion capacity, and are in accord with the international quality control standards of the mesenchymal stem cells.Meanwhile, according to the culture method disclosed by the invention, a large number of mesenchymal stem cells (particularly human umbilical cord mesenchymal stem cells) can be obtained by amplifyinga small number of mesenchymal stem cells under the conditions of small space occupation, less culture medium consumption, simpler and more convenient operation and greatly reduced workload; and a solid technical scheme and a theoretical basis are provided for obtaining a large number of high-quality mesenchymal stem cells (especially human umbilical cord mesenchymal stem cells) and secretions thereof in the clinical field.

A serum-free cartilage cell culture solution is obtained by adding exogenous additives to a basic culture solution. The basic culture solution is a DMEM culture solution or an F12 culture solution or a DMEM culture solution-F12 culture solution mixed culture solution, and the exogenous additives are nonessential amino acids, glucan, vitamin C, glutamine, sodium pyruvate, insulin or insulin-like growth factor, transferrin, growth hormone, triiodothyronine, hydrocortisone, dexamethasone, sodium selenite, beta-mercaptoethanol, lipid concentrate, progesterone, succinimide, a basic fibroblast growth factor, an epidermal growth factor, a transforming growth factor, a bone morphogenetic protein and a platelet derived growth factor respectively. The serum-free cartilage cell solution can avoid potential risks due to the use of serum, improves the extracellular matrix secreting ability of cartilage cells, makes the growth propagation state approach the growth propagation state of the cells in a serum-containing culture solution, and allows the dedifferentiation rate and the matrix secreting ability of low density inoculation cartilage cells to be obviously better than those of the cells in the serum-containing culture solution respectively.

The invention provides alginate lyase, host cells capable of secreting alginate lyase and application of the alginate lyase and the host cells. The alginate lyase has any one of amino acid sequences shown as (I) and (II): (I) an amino acid sequence shown as SEQ ID NO. 1; (II) an amino acid sequence obtained by carrying out substitution, deletion or addition of 1 to 20 amino acid residues on the amino acid sequence shown as SEQ ID NO. 1; the obtained amino acid sequence has the activity of the alginate lyase. According to the alginate lyase, whole genes of the alginate lyase are truncated to reduce the molecular weight, so that the expression amount of the host cells on the alginate lyase is remarkably improved; the secreted alginate lyase has high activity, good stability and strong degradation capability on sodium alginate.

The invention discloses a recombinant duck interleukin-2 protein preparing method and its use, the DNA fraction of RT-PCR amplified duck interleukin-2c of different ducks and prokaryotic expression carrier pBAD/HisB, as well as eukaryotic expression carrier pMET alpha A to compose high-efficacy expression particles, recombining and converting these carriers to Escherichia coli LMG194, and microzyme strains PMAD11 and PMAD16, respectively and then inducing expression. After inducing, making ultrasonic wave cracking and deposit elimination by centrifugation on Escherichia coli to obtain crude products of duck interleukin-2 and using protein purifying system to obtain duck interleukin-2 pure products. The pure or crude of duck interleukin-2 can act as immune assistant, anti-disease additive and disease-curing drug. The duck interleukin-2 monoclonal and polyclonal antibodies have simple preparing procedure, and can act as good immune inhibitor. It applied genetic engineering technique to prepare recombinant protein gsIL-2, and has simple technical flow, low production cost, good stability, high bioactivity and other characters.

The invention relates to the technical field of thelephora ganbajun Zang polysaccharides, in particular to a thelephora ganbajun Zang exopolysaccharide. A preparation method of the thelephora ganbajun Zang exopolysaccharide includes the steps: fermenting thelephora ganbajun Zang strains TG-1; extracting the thelephora ganbajun Zang exopolysaccharide from fermentation broth; separating mixture by the aid of an anion exchange column chromatography; eluting by the aid of deionized water, 0.1 mol/L of NaCl solution and 0.3 mol/L of NaCl solution to respectively obtain thelephora ganbajun Zang exopolysaccharide components such as EPS-1, EPS-2 and EPS-3. The preservation number of the thelephora ganbajun Zang strains TG-1 is CGMCC No.12977. The thelephora ganbajun Zang EPS has good humidity preservation and antioxidant activity, antioxidant ability of the EPS-2 in the three components is highest, and the EPS is high in the humidity preservation ability. The thelephora ganbajun Zang exopolysaccharide can be widely applied to anti-aging health care products and cosmetics.

The invention discloses a preparation method of oxalate decarboxylase. The method comprises the following steps: constructing a recombinant expression plasmid vector comprising Bacillus-containing acid induced promoter, a secretion signal peptide, and a fungus-derived oxalate decarboxylase gene prepared under the control of the secretion signal peptide, transforming bacilli normally growing in a pH value being not more than 4.5 by the vector to prepare oxalate decarboxylase recombinant bacteria; and carrying out culture and acid feeding on the recombinant bacteria in a fermentation medium, inducing the recombinant bacteria, and carrying out simple purification to recover generated extracellular oxalate decarboxylase. The extracellular oxalate decarboxylase prepared through the method is used in diagnosis reagent raw materials, medicinal compositions, healthcare foods or formulated foods with special medical uses. The mutated acid induced promoter is adopted to substitute an original acid induced promoter in the recombinant expression vector, so the transformed bacilli have a high expression level.

The invention belongs to the technical field of medical treatment and health and particularly relates to a concentrated adipose-derived stem cell culture supernatant preparation to treat skin scalding. The concentrated adipose-derived stem cell culture supernatant preparation is prepared with supernatant of a cell culture fluid cultured via adipose-derived mesenchymal stem cells; cell factors hrEGF, VEGF and bFGF are added to the preparation. The invention also provides a preparation method and application of the concentrated adipose-derived stem cell culture supernatant preparation. The concentrated adipose-derived stem cell culture supernatant preparation of the invention can significantly promote healing of scald wounds, and the results lie in the aspect: after the use of the preparation, epidermis and dermis can heal and restore their functionality faster than those treated in a positive control and other embodiment groups. In addition, the concentrated adipose-derived stem cell culture supernatant preparation of the invention is free of living cell ingredients, no ethic issues or immune safety issues occur, compounding the preparation when it is needed is not required, and thepreparation is convenient to apply.

The invention relates to mesenchymal stem cell composite active factor freeze-dried powder as well as a preparation method and application thereof. In the preparation process of the composite active factor freeze-dried powder, a specific pre-coating solution is used for treating a cell culture container; and mesenchymal stem cells are cultured by adopting a first culture medium and a second culture medium under specific culture conditions, so the mesenchymal stem cells can be still kept at a normal state in the processes of hunger culture and induction and secrete more active factors. The composite active factor freeze-dried powder prepared by the invention can promote cell division and cell proliferation, repair damaged skin or skin with thin congenital cuticle, tighten the skin and reduce wrinkles; moreover, the immunoregulation function of the skin is activated, the metabolism of the skin is promoted, and an anti-aging effect is achieved.

The invention belongs to preparation of a new gene and construction of an engineering bacterium, and particularly relates to a neutral proteinase gene constructed by a molecular chaperone prsA, a protein, bacillus subtilis, preparation and an application. The molecular chaperone PrsA and the neutral proteinase gene are integrated to construct pHT43-npr-PrsA recombinant plasmid by a molecular biology technology and finally converted into a host bacterium, namely the bacillus subtilis WB800N, to construct a new bacillus subtilis engineering bacterium CGMCC No. (China General Microbiological Culture Collection Center number) 14837; the molecular chaperone PrsA and the neutral proteinase gene are co-expressed; and an expression level of neutral proteinase is effectively increased.

The invention discloses recombinant bacillus subtilis of exocytosis PNAG polysaccharide. A PNAG polysaccharide synthetase coding gene and expression plasmid are connected, recombinant expression plasmid is constructed, then the recombinant expression plasmid is transferred into bacillus subtilis, and the recombinant bacillus subtilis is obtained. The invention further discloses a construction method of the recombinant bacillus subtilis. A new metabolism synthetase gene is guided into bacillus subtilis, the recombinant bacillus subtilis is successfully constructed, PNAG polysaccharide can be produced through fermentation, the produced exocytosis PNAG polysaccharide can reach 136mg/L, and a base is established for further reforming the bacillus subtilis for producing the PNAG polysaccharidethrough metabolic engineering. The invention further discloses an application of the recombinant bacillus subtilis to preparation of products containing the PNAG polysaccharide through fermentation indifferent fields of foods, medicines and chemical industry. The PNAG polysaccharide produced from recombinant strains can promote growth of probiotics and restrain harmful bacteria, and has good application prospects in many fields.

The invention relates to a recombinant pichia pastoris strain for regulating and controlling transglutaminase expression of zea mays through a TEF1 promoter. The recombinant pichia pastoris strain isa genetically engineered strain obtained in the mode that transglutaminase from the zea mays is transferred to pichia pastoris. According to the recombinant pichia pastoris strain, an alcohol oxidasepromoter PAOX1 in a pPIC9K plasmid is replaced by a promoter PTEF1 of a translation extension factor 1-alpha, the promoter PTEF1 is connected with a transglutaminase gene to form a loop, and a recombinant expression vector is constructed. The recombinant strain is easy to culture, high in secretion capacity and beneficial for mass transglutaminase expression, the enzyme activity can reach 479 mU/mL, and the recombinant pichia pastoris strain has the advantages of being safe and reliable and has important application prospects.

The invention belongs to the field of biological medicine, and relates to an anti-TIGIT antibody and application thereof. Specifically, the invention relates to an anti-TIGIT antibody or an antigen binding fragment thereof, the antibody comprises a heavy chain variable region and a light chain variable region, the amino acid sequences of HCDR1 to HCDR3 of the heavy chain variable region are respectively shown as SEQ ID NOs: 3-5, and the amino acid sequences of LCDR1 to LCDR3 of the light chain variable region are respectively shown as SEQ ID NOs: 8-10; and according to an EU numbering system, the heavy chain constant region comprises mutation such as L234A and L235A. The antibody disclosed by the invention can be effectively combined with TIGIT, has small toxic and side effects and has the potential of being applied to tumor prevention and treatment.

The invention relates to the technical field of microorganisms and biology, in particular to aspergillus sydowii as well as a separation method and application thereof. The aspergillus sydowii JDQMZZ-1 has been deposited in China General Microbiological Culture Collection Center on March 12, 2020 with a deposit number of CGMCC No.19278, and has nitrogen fixation and strong secretion ability; has a strong inhibitory effect on fungi and bacteria, can also promote germination of plant seeds, and can achieve growth-promoting effects by adjusting a content of plant endogenous hormones; can promote growth of underground part of seedlings of Paris polyphylla after applying to the seedlings of Paris polyphylla, but does not activate plant autoimmunity, and effectively avoids a result of activating plant autoimmunity leading to reduced biomass. Meanwhile, the aspergillus sydowii JDQMZ-1 increases chlorophyll content of plants and reduces malondialdehyde content, which indicates that when the aspergillus sydowii JDQMZ1 acts on seedlings of Paris polyphylla, an influence of the environment on the seedlings of Paris polyphylla is reduced, damage to cell membranes is reduced, and abiotic stress resistance of the seedlings of Paris polyphylla is improved.

The invention discloses a method for preparing stomach-regulating pills, which comprises the following steps of: respectively performing volatile oil extraction, water temperature extraction and alcohol extraction on different medicinal materials in the stomach-regulating formula, extracting effective parts therein, and mixing to obtain an intermediate for preparing the pills; and mixing the intermediate and a substrate, dispersing into the substrate, and performing a pill forming process to obtain the stomach-regulating pills. In the pill preparing process, each extraction link and the forming process have controllable quality, the content of active ingredients is stable, the pill preparation is carried out after the medicament intermediate is mixed fully with the substrate, and the medicament active ingredients are highly dispersed, so the prepared pills have high medicament stability.

The invention relates to the technical field of pearl cultivation, in particular to a novel method for cultivating large free pearls by pinctada maxima. The novel method mainly comprises the following steps of: cutting off 30-40% of retractor of pinctada maxima pearl cultivating shells, reducing the extrusion strength of the retractor on pearl nucleuses, implanting only one perfect circular pearl nucleus at a left bag nucleus position for the first nucleus implantation, and during the second and third nucleus implantation, implanting one pearl nucleus at the left bag nucleus position and a right bag nucleus position respectively. The pinctada maxima belongs to rare protected species, in order to make full use of the pearl cultivation potential of the pinctada maxima, nucleus implantation is carried out three times in total, and meanwhile, the diameter of harvested pearls is 2.0-3.0 mm larger than that of pearls obtained in the previous step every time nucleus implantation is added; and by adopting the nucleus implantation cultivation method, the pearl cultivation capacity of the pinctada maxima is utilized to the maximum extent, the survival rate, the nucleus remaining rate and the superior pearl rate of the pinctada maxima are increased, and large free pearl industrial cultivation of the pinctada maxima can be achieved.

The invention discloses an SD rat-derived mesenchymal stem cell induction medium and induction method. The induction medium is composed of an alpha-MEM medium and an additive, wherein the additive is composed of 5-100 ng/ml of a transforming growth factor-alpha, 10-200 ng/ml of regulatory protein [beta]1, 100-3000 nM of trehalose and 50-5000 nM of dibutyryl cyclic adenosine monophosphate according to the final concentration. According to the SD rat-derived mesenchymal stem cell induction culture medium and induction method, the secretion amount of factors such as BDNF, HGF, VEGF and GDNF of rat mesenchymal stem cells can be obviously increased, and the secreted factors are relatively rich in variety and can be stably secreted for a long time; and the risks and uncertainty caused by other methods such as gene modification are avoided through a cytokine induction method, and the induction method is more suitable for industrialization and clinical application.

The invention discloses a high-yield recombinant strain of trichoderma harzianum alpha-1, 3-glucanase and an application thereof. The high-yield recombinant strain of trichoderma harzianum alpha-1, 3-glucanase and the application thereof are characterized in that alpha-1, 3-glucanase genes from trichoderma harzianum CCM F-340 conduct codon optimization; with pyr4 genes used as screening marker genes, trichoderma reesei cbh1 strong promoter and signal peptide sequence thereof and cbh1 terminator are used to construct alpha-1, 3-glucanase expression vector pSK-mutAW; highly secreted expression of the trichoderma harzianum alpha-1, 3-glucanase in trichoderma reesei is achieved. Experiments show that the high-yield recombinant strain of trichoderma harzianum alpha-1, 3-glucanase and the application thereof have the advantages that recombinant expression of the trichoderma harzianum alpha-1, 3-glucanase can effectively degrade non-water-soluble glucan mutan with fermentation enzyme activityunit reaching 2.3 milliliters and alpha-1, 3-glucanase content in fermentation broth being about 68 micrograms per milliliter; fermentation process and target protein purification process of the sameare simple and very suitable for industrial production and application.