1511results about "Plant cells" patented technology

The invention discloses an extraction and purification method and application of plant-derived exosome-like nanoparticles, and relates to the technical field of plant exosome preparation. The extraction and purification method comprises the following steps: washing fresh ginseng, airing for later use, adding a proper amount of buffer solution, mixing, breaking walls, filtering, centrifuging filtrate to obtain supernate, carrying out ultracentrifugation on the supernate to obtain ginseng exosome nano-particle precipitates, resuspending the precipitates, respectively adding pure water sucrose solutions with different concentrations, and carrying out ultracentrifugation to obtain ginseng exosome nano-particles; and re-suspending the target strip, and carrying out ultracentrifugation to remove redundant cane sugar so as to obtain the purified ginseng exosome-like nano-particles. The ginseng exosome-like nano-particles disclosed by the invention have a remarkable effect when being applied to cosmetics or medicines with skin photoaging resistance, skin oxidation resistance and wrinkle resistance. In an anti-UVB (Ultraviolet B) induced cell injury experiment, the ginseng exosome-like nano-particles have an excellent anti-UVB induced cell injury effect.

The invention discloses a method for extracting extracellular vesicles from dried Sanguisorba officinalis rhizomes and application of the extracellular vesicles, and relates to the technical field of molecular biology and biomedicine.The method comprises the following steps that S1, rehydration is conducted, specifically, the dried Sanguisorba officinalis rhizomes are subjected to liquid nitrogen quick freezing and then ground into powder at the low temperature, the powder and hydration liquid are mixed according to the proportion of 1: 10 (w / v), the mixture is slowly stirred for 2-4 h at the temperature of 4 DEG C, and the extracellular vesicles are obtained; removing large-particle impurities by using sterile gauze or a filter screen, and collecting filtrate; according to the method, through the synergistic effect of multiple enzymolysis and in combination with cellulase, pectinase and the like, the degradation efficiency is remarkably improved, meanwhile, EDTA is chelated with metal ions, and the enzymolysis effect is enhanced. Short-time alternating ultraviolet light is introduced to activate'dormant 'vesicles, so that the yield and the biological activity are improved. Secondly, multiple protection mechanisms, such as vitamin C, EDTA (Ethylene Diamine Tetraacetic Acid), cane sugar, BSA (Bovine Serum Albumin) and an ultraviolet protective agent are adopted, so that the integrity and the biological activity of the vesicles are ensured.

The invention discloses a ginseng exosome, an extraction method thereof and application of the ginseng exosome in preparation of anti-photoaging skin care products, and belongs to the technical field of biological medicines. In order to solve the technical blank of ginseng exosomes in the aspect of light aging resistance in the prior art and solve the technical problems of complicated steps, long time consumption and small treatment capacity of exosome extraction by an ultracentrifugation method in the prior art, the invention provides an extraction method of ginseng exosomes. Therefore, the extraction rate of the ginseng exosome is improved, the treatment steps are simplified, the time is saved, and the purpose of producing the ginseng exosome in a large scale is achieved. The ginseng exosome obtained through the extraction method can be applied to preparation of anti-photoaging skin care products, and clinical application of the exosome is achieved.

The invention discloses application of camellia nitidissima exosome-like nano-vesicles in preparation of antitumor drugs, and belongs to the technical field of biological medicines. The exosome-like nano-vesicles are extracted from golden camellia for the first time, and the exosome-like nano-vesicles are proved to have remarkable anti-tumor activity.

The invention relates to polygonum multiflorum exosome-like vesicles and application thereof, the polygonum multiflorum exosome-like vesicles are applied to preparation of antioxidant products, anti-inflammatory products and anti-ultraviolet products, the antioxidant products are products for removing one or more of DPPH free radicals, ABTS free radicals and hydroxyl free radicals, the anti-inflammatory products are products for inhibiting hyaluronidase, and the anti-ultraviolet products are products for removing one or more of DPPH free radicals, ABTS free radicals and hydroxyl free radicals. And the anti-ultraviolet product is a product capable of absorbing UVB. Compared with the prior art, the polygonum multiflorum exosome-like vesicles prepared by the method disclosed by the invention are low in cost and large in treatment capacity, and the obtained polygonum multiflorum exosome-like vesicles have excellent oxidation resistance, inflammation resistance and ultraviolet resistance.

The embodiment of the invention discloses a method for extracting extracellular vesicles from a traditional Chinese medicine dry product. The method comprises the following steps: taking a dry traditional Chinese medicine product as a raw material, and sequentially performing crushing, soaking, first-order enzymolysis, wall breaking, differential centrifugation, second-order enzymolysis, high-speed centrifugation, ultracentrifugation and ultrafiltration enrichment to obtain the extracellular vesicles. A two-step enzymolysis method is adopted in the enzymolysis process, impurities are removed to the maximum extent in a sequence difference enzymolysis mode, and the structure and function stability of the exosome are protected. According to the invention, the differential centrifugation and the enzymolysis method are alternately used, so that impurities with different sizes and different properties can be removed more thoroughly. According to the method, the cost is strictly controlled, the exosome purification efficiency is relatively high, the concentration of the obtained dry exosome is relatively high, and the operability is extremely high.

The invention discloses a preparation method and application of plant-derived extruded nano-vesicles, and relates to the technical field of biological medicines. The preparation method of the plant-derived extruded nano-vesicles comprises the following steps: juicing cleaned plant roots, stems, leaves, flowers or fruits to obtain plant juice, and filtering with gauze to obtain first filtrate; centrifuging the first filtrate at a low speed of 300-6000 * g to obtain a second filtrate; performing 8,000-20,000 * g centrifugation on the second filtrate for 0.5-3 hours at the temperature of 0-5 DEG C, so as to obtain a first precipitate; adding the first precipitate into a buffer solution, carrying out ultrasonic treatment, diluting, and sequentially passing through a filter membrane with the pore diameter of 0.4-10 microns and a filter membrane with the pore diameter of 0.1-0.4 microns, so as to obtain a third filtrate; and centrifuging the third filtrate, taking the precipitate, and resuspending to obtain the extruded nano-vesicles. Experimental detection shows that the extruded vesicles are similar to natural vesicles in characteristics, but the yield is far higher than that of the natural vesicles, and the extruded vesicles can serve as nano-carriers to be applied to a foundation or clinic.

The present invention relates to a method for extracting exosome-like nanovesicles from Platycladus orientalis leaves by freeze-drying and enzymatic hydrolysis, and the extracted exosome-like nanovesicles and applications thereof. Specifically, the freeze-drying and enzymatic hydrolysis method for extracting exosome-like nanovesicles from Platycladus orientalis leaves provided by the present invention ensures a stable supply of raw materials, is easy to store for a long time, and significantly improves the extraction efficiency and economy, opening up a new path for the commercial production of exosome-like nanovesicles from plant sources. Through freeze drying and low-temperature enzymatic hydrolysis technology, the active ingredients in Platycladus orientalis leaves are retained to the maximum extent, and the fiber and polysaccharide colloid are effectively removed, thereby improving the purity and biological activity of the exosome-like nanovesicles. The method is simple, easy to operate, low in cost, and suitable for large-scale production. In addition, the present invention applies the extracted Platycladus orientalis leaves exosome-like nanovesicles to hair growth, especially for androgenic alopecia (AGA), providing a safe and efficient new treatment strategy, and is expected to lead a new trend in the preparation and application of plant exosome-like nanovesicles.

The invention discloses a method for efficiently separating and purifying plant exosomes and application, and belongs to the technical field of biological nanotechnology. The method comprises the following key steps: (1) performing sterile cleaning on plant raw materials, and then performing peeling or tissue slitting pretreatment; (2) carrying out mechanical crushing treatment by adopting a phosphate buffer solution (PBS), and filtering through a 200-mesh screen to obtain a primary extracting solution; (3) performing gradient centrifugal purification on the extracting solution, collecting supernate, and filtering the supernate through a 0.45 mu m microporous filter membrane; (4) adding 8-12% of polyethylene glycol for low-temperature phase separation and precipitation; (5) purifying by using an electrodialysis technology; and (6) carrying out final concentration by adopting a 100kDa ultrafiltration membrane to obtain the high-purity PELNs preparation. According to the method, the process, equipment and reagents are simple and easy to obtain, the final yield is high, the purity is high, particle size distribution is uniform, and an innovative solution is provided for large-scale industrial production of natural-source nano-drug carriers.

The invention relates to a Chinese herbal medicine extracellular vesicle composition and a preparation method and application thereof, and the Chinese herbal medicine extracellular vesicle composition comprises at least four of turmeric extracellular vesicles, polygonum cuspidatum extracellular vesicles, polygonum multiflorum extracellular vesicles, cacumen biotae extracellular vesicles and dried ginger extracellular vesicles. Wherein the Chinese herbal medicine extracellular vesicle composition contains turmeric extracellular vesicles. The Chinese herbal medicine extracellular vesicle composition provided by the invention has the effects of growing hair and preventing hair loss, is safe and free of toxic and side effects, and can be easily taken by cells.

The invention relates to the field of biological medicine, and discloses a rhizoma polygonati-fructus lycii heterozygous exosome, a preparation method and application thereof and an antioxidant composition. The hybrid exosome comprises miRNA-156a (micro Ribonucleic Acid) and miRNA-396b (micro Ribonucleic Acid), and the expression quantity ratio of the miRNA-156a to the miRNA-396b is (1.2 to 1.8): 1; the heterozygous exosome is loaded with a polygonatum kingianum characteristic active component and a Chinese wolfberry fruit characteristic active component. The rhizoma polygonati-fructus lycii heterozygous exosome is good in biocompatibility and capable of remarkably improving the cell survival rate, and brand-new and efficient raw materials and technical supports are provided for development of anti-aging and anti-oxidation functional products.

The present disclosure is in the field of plant genetic engineering. Specifically, the invention relates to application of a mutant GmLn protein and an encoding gene thereof in regulation and control of soybean yield, in particular to but not limited to a method for obtaining soybeans capable of being densely planted, a method for detecting the soybeans capable of being densely planted, related primers and kits, and further relates to soybean plants capable of being densely planted and a method for improving soybean planting density. According to the method provided by the invention, the soybean target sequence is subjected to gene editing by expressing the gene editing tool in the soybean, so that the soybean which is improved in yield and can be densely planted is obtained.

The invention discloses a preparation method of rhizoma polygonati exosome microcapsules, and belongs to the technical field of biology, the rhizoma polygonati exosome microcapsules are prepared by four steps of crude extraction of rhizoma polygonati exosome, purification of the rhizoma polygonati exosome, preparation of rhizoma polygonati exosome freeze-dried powder and preparation of the rhizoma polygonati exosome microcapsules. The rhizoma polygonati exosome is wrapped in the tiny vesicles formed by the high polymer material, so that the external environment is effectively isolated, a protective barrier is provided for the exosome, the exosome is prevented from being degraded or inactivated in the external environment, and the stability of the exosome is improved; meanwhile, the polygonatum kingianum exosome can be prevented from being subjected to in-vivo enzymolysis and clearing through micro-capsule wrapping, the in-vivo residence time of the polygonatum kingianum exosome is prolonged, and the bioavailability is improved; the preparation process is simple, the operability is high, and the prepared exosome microcapsule has good application in the aspect of inhibiting gastric cancer cells.

The invention relates to application of a tea exosome in preparation of a medicine for improving skin pigmentation, the tea exosome is creatively extracted and purified, and the tea exosome can effectively inhibit proliferation of melanoma cells and inhibit expression of melanin generation related protein by scavenging free radicals and inhibiting tyrosinase activity, so that the effect of improving skin pigmentation is achieved. The intracellular melanin removal is promoted. In a mouse in-vivo experiment, after the tea exosome is applied, it is observed that the skin epidermis thickness of a mouse is reduced, the dermis thickness is increased, collagen arrangement is tighter and more ordered, the content is increased, and meanwhile generation of inflammatory factors is reduced. The tea exosome can significantly increase the content of a melanin autophagy key factor LC3B and reduce the content of P62 to promote melanin autophagy, and has good percutaneous infiltration capacity, thereby playing a role in improving skin pigmentation.

The invention relates to Chinese pulsatilla root and stem exosome-like vesicles as well as a preparation method and application thereof. The preparation method comprises the following steps: juicing Chinese pulsatilla roots, filtering, and collecting filtrate; and centrifuging the filtrate, collecting the precipitate, re-suspending, and filtering to obtain a solution containing the exosome-like vesicles of the Chinese pulsatilla roots. Compared with the prior art, the Chinese pulsatilla root and stem exosome-like vesicles extracted by the method can inhibit malignant progression, including proliferation and migration, of lung cancer, and cell uptake and animal living imaging prove that the Chinese pulsatilla root and stem exosome-like vesicles can be taken by cells and can be enriched in tumor tissues at the same time.

The invention relates to a method for preparing a Chinese wolfberry exosome, the Chinese wolfberry exosome prepared by the method and application of the Chinese wolfberry exosome. Specifically, the preparation method of the lycium barbarum plant exosome disclosed by the invention comprises a layer-by-layer filtration step, and the layer-by-layer filtration step sequentially comprises deep filtration and optional membrane filtration.

The invention relates to the technical field of plant exosome-like extracts, in particular to a liquorice exosome-like extract as well as extraction and application thereof. The method comprises the following steps: firstly, removing impurities such as cells, cell debris, organelles, large vesicles and the like in homogenate by adopting a combination mode of low-speed centrifugation and medium-speed centrifugation, and further purifying the homogenate by adopting high-speed centrifugation after the low-speed centrifugation and the medium-speed centrifugation are completed, so as to obtain the exosome-like extract. And the setting of the increasing rotation speed is beneficial to finer separation of exosome-like particles with different sizes and densities, so that the purity and quality of the extract are improved. According to the method, all centrifugal operations are carried out in a low-temperature environment of 4 DEG C; the low-temperature condition is beneficial to maintaining the biological activity and stability of the sample, and the problems of protein denaturation, enzyme inactivation and the like caused by temperature rise are reduced, so that the quality and performance of the exosome-like extract are ensured.

The invention belongs to the technical field of preparation of anti-tumor drugs, and particularly relates to a coptis root exosome, an extraction method thereof and application of the coptis root exosome in tumor resistance. The coptis root exosome is extracted from coptis roots, the size of the coptis root exosome is 125nm-135nm, and the coptis root exosome is shaped like a vesicle with a lipid bilayer structure. The coptis root exosome has an excellent anti-tumor effect and can remarkably inhibit tumor growth, and the tumor growth inhibition effect is remarkably superior to that of a common extract berberine in coptis roots.

The invention discloses a fruit and vegetable exosome extraction method based on ligand fishing. The fruit and vegetable exosome extraction method comprises the following steps: suspending amino-functionalized Fe3O4 (at) SiO2 nanoparticles in a glutaraldehyde solution, and adding activated agglutinin to obtain Fe3O4 (at) SiO2-agglutinin nanoparticles; adding a glutaraldehyde solution into agarose gel to obtain a size exclusion chromatographic column filler; the method comprises the following steps: dispersing Fe3O4 (at) SiO2-lectin nanoparticles in a PBS buffer solution to obtain a uniform suspension, adding the uniform suspension into a size exclusion chromatographic column filler to obtain a uniform slurry, filling the size exclusion chromatographic column with the uniform slurry, and flushing with the buffer solution to form a magnetic bead stationary phase; and loading fruit and vegetable supernatant into a size exclusion chromatographic column, passing through the magnetic bead stationary phase, cleaning residual impurities combined on the exosome-magnetic bead compound by using a buffer solution, and dissociating the exosome from the magnetic beads by using a competitive eluent of the exosome to obtain the high-purity fruit and vegetable exosome. The problem that an existing exosome extraction method is difficult to meet the requirements of high efficiency, simplicity, high purity and high activity at the same time is solved.

The invention discloses a large-scale preparation method of ginseng exosomes, which comprises the following steps: culturing calluses: inoculating explants into an induction culture medium until ginseng calluses are generated; proliferation of the calluses: inoculating the newborn calluses into a proliferation culture medium for proliferation culture; inducing germination and rooting: after the ginseng callus is amplified, transplanting the ginseng callus into a germination induction culture medium until the ginseng callus is germinated, and then transferring the ginseng callus into a rooting culture medium for culturing and rooting; and preparing the exosome, namely grinding root hairs grown from the ginseng callus regeneration plant, uniformly mixing the ground root hairs with PBS (Phosphate Buffer Solution) or performing liquid culture on ginseng suspension cells, and finally performing centrifugal treatment to obtain the ginseng exosome. According to the large-scale preparation method of the ginseng exosome, the endotoxin pollution in the growth and development process is reduced through ginseng callus culture rooting, and then the ginseng exosome is prepared, so that the large-scale preparation requirement in modern production and processing is met.

The invention relates to a method for producing notoginsenoside and ginsenoside by utilizing panax notoginseng cotyledon embryos, and belongs to the technical field of plant adventitious root tissue culture. The method comprises the following steps: inducing embryogenic calluses by utilizing immature zygotic embryos of panax notoginseng; proliferating the panax notoginseng embryogenic callus on a plate culture medium; s2, putting the pseudo-ginseng embryogenic callus obtained in the step S2 into a liquid culture medium, carrying out cell dispersion, and sieving by using a sieve with the aperture of 24-30 meshes; a mechanical stirring type bioreactor is filled with a liquid culture medium, and the globular embryo and the heart-shaped embryo are placed in the mechanical stirring type bioreactor to be cultured; s3, recovering the filtrate in the step S3, filtering by using a screen with the aperture of 30-80 meshes and 80-150 meshes to obtain uniform panax notoginseng embryogenic cells, and differentiating the panax notoginseng embryogenic cells in a triangular flask for the step S4; and drying the cotyledon embryo obtained in the step S4 at 45 DEG C, and grinding into powder. According to the method, large-scale generation of cotyledon embryos can be completed only in 4-6 weeks, the production process is remarkably accelerated, the method is suitable for industrial batch production requirements, and the production efficiency is high.

The invention discloses a tissue culture method of a double-petal hemerocallis fulva scapes, which comprises the following steps: taking a double-petal hemerocallis fulva scapes, taking a growing point as a center, transversely cutting into long sections which are not less than 5cm, washing and disinfecting; putting the disinfected long segments into enzymatic hydrolysate for cell wall enzymolysis, taking out the long segments after enzymolysis is completed, and washing the long segments with sterile water; transversely cutting the long section subjected to enzymolysis into small sections of 1-2cm by taking the growing point as the center; and carrying out tissue culture by taking the small sections as explants. The method applies the double-petal hemerocallis fulva variety, and has the advantages that materials can be taken at any time in the flowering phase, the tissue culture efficiency is high, and the quality is good.

The invention discloses an implant-derived exosome which is prepared by the following steps: adding cleaned and crushed colored rice flour into a PBS (Phosphate Buffer Solution) with the pH value of 6-7.4, homogenizing at a high speed, adding alpha-amylase into rice milk, performing enzymolysis for 1-2 hours under the conditions that the pH value is 6-7.4 and the temperature is 90-97 DEG C, filtering, and standing filtrate overnight; carrying out gradient differential centrifugation at 4 DEG C, collecting supernate, filtering, carrying out ultracentrifugation on filtrate at 4 DEG C and 12000-15000g for 1.5-3 hours, and collecting precipitate; and re-suspending the precipitate, enriching by using an MWCO ultrafiltration tube with the molecular weight cut-off of 100kDa, collecting the precipitate at the bottom of the ultrafiltration tube, and re-suspending in a PBS buffer solution, thereby obtaining the product. The exosome can effectively inhibit the generation of inflammatory factors, has a good inhibition effect on various pathogenic bacteria, also has an anti-oxidation effect, and has application prospects in the fields of inflammation resistance, bacteria resistance, oxidation resistance and the like.

The invention relates to the technical field of freeze-dried powder preparation, and discloses a white tomato exosome freeze-dried powder preparation method, which comprises: 1, manually picking white tomatoes 3-5 days after a color changing period, carrying out double-frequency ultrasonic cleaning, and pre-cooling in a 4 DEG C phosphate buffer solution containing 0.1% of vitamin C; according to the method, by virtue of an ultrasonic-cellulase synergistic crushing process, cells are mildly and efficiently crushed, so that the damage to an exosome structure is reduced, and the release efficiency and activity of the exosome are improved; the zebra fish embryo melanogenesis can be obviously inhibited, and the whitening and freckle-removing effects are relatively good.

The invention discloses a preparation method of artemisia apiacea extracellular vesicles, which comprises the following steps: step 1, pre-treating artemisia apiacea, carrying out primary centrifugal treatment, removing large plant tissues and cell debris, carrying out secondary centrifugal treatment, filtering by a needle filter, and collecting; step 2, transferring the filtered suspension to a sucrose density gradient solution, then carrying out third centrifugal treatment, collecting the solution with the concentration of 30% in the layer, and carrying out fourth centrifugal treatment to obtain artemisia apiacea extracellular vesicles; the invention further discloses an artemisia apiacea extracellular vesicle, a pharmaceutical composition for treating cerebral apoplexy, a medicine and application of the artemisia apiacea extracellular vesicle in preparation of the medicine for treating cerebral apoplexy. The naturally-sourced vesicle delivery system provided by the invention is expected to have higher safety and lower toxic and side effects, and the enrichment concentration of the therapeutic component at the focus part of cerebral apoplexy is improved, so that the bioavailability and the treatment efficiency of the medicine are remarkably improved.

The present invention provides a method for producing extracellular vesicles, the method including a step for passing a crushed liquid of plants, mushrooms, or microalgae through a depth filter, and a step for subjecting the crushed liquid after passing through the depth filter to ultrafiltration treatment, the depth filter being composed of filaments.

The invention particularly relates to preparation of grape exosomes or a composition thereof and application of the grape exosomes or the composition in skin medical treatment and beauty and health care. Skin cells Hacat are used as a model, and it is verified that the grape exosome can significantly reduce the active oxygen level in the skin cells Hacat, inhibit expression of inflammatory factors TNF-alpha and IL-1beta, significantly promote expression of Collagen1, Elastin, FLG and LOR in the skin cells Hacat, inhibit inflammatory factors TNF-alpha and IL-1beta, inhibit inflammatory factors TNF-alpha and IL-1beta, inhibit inflammatory factors TNF-alpha and IL-1beta, inhibit inflammatory factors TNF-alpha and IL-1beta and inhibit inflammatory factors TNF-alpha and IL-1beta from inflammatory factors TNF It is proved that when the grape exosome acts on skin cells, the effects of resisting inflammation, resisting oxidation, promoting skin scar healing and tightening and resisting wrinkles are expected to be achieved at the same time, and the application prospect in the aspect of medical cosmetology is huge.

The invention discloses a preparation method of a raspberry-derived exosome loaded with neroli essential oil. The preparation method comprises the following steps: preparing raspberry fruits into homogenate; centrifuging, collecting supernate, adjusting the pH value to 4.07.5, standing, and centrifuging again; taking supernate, adjusting the pH value to 7.2-7.4, adding polyethylene glycol, standing, centrifuging, collecting precipitate, and carrying out resuspension dispersion; performing microfiltration to obtain supernate, and combining electrophoresis dialysis and ultrafiltration concentration to obtain the high-purity raspberry exosome. And diluting the exosome, adding the diluted exosome into a dimethyl sulfoxide solution containing neroli essential oil, carrying out water bath stirring, carrying out ultrafiltration centrifugation, and taking a clear liquid to obtain the exosome loaded with the neroli essential oil. Efficient separation and purification of the raspberry exosome are achieved by applying an isoelectric precipitation technology, then the neroli essential oil is wrapped in the exosome, the stability of the essential oil is improved, the action time is prolonged, meanwhile, the neroli essential oil and active ingredients of the exosome generate a synergistic effect, and the antioxidant activity is jointly enhanced.

The invention provides an exosome enrichment method based on a modified cellulose material, which comprises the following steps: carrying out TEMPO oxidation on plant residue cellulose to prepare a cellulose material with a carboxyl structure; the cellulose structure is further refined by adopting mechanical shearing and high-pressure homogenizing means, so that the specific surface area and the dispersion stability are remarkably improved; through polyethyleneimine PEI grafting, a large number of amino functional groups are introduced, the surface electropositivity of the material is enhanced, and efficient adsorption and enrichment of exosomes with negative charges are achieved. The invention also provides the exosome prepared by the method. The modified cellulose material disclosed by the invention is wide in source, simple and convenient to prepare and mild in enrichment process, the separation purity and the enrichment efficiency of the exosome can be remarkably improved, and the modified cellulose material has good biocompatibility and environmental friendliness; compared with existing ultracentrifugation, density gradient centrifugation and immunoaffinity separation methods, the method has the advantages that the operation process is simplified, the equipment dependence and reagent cost are reduced, and the potential of large-scale production and clinical application is achieved.