2023results 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 relates to a universal nano biological vesicle preparation method. The preparation method comprises the following steps: carrying out crude extraction on cells from bacteria, fungi, plants or animals to obtain a crude extracting solution; the crude extracting solution is placed in a low-temperature and high-pressure homogenizer to be subjected to low-temperature and high-pressure homogenizing treatment, then the nano vesicles are obtained, the low-temperature and high-pressure homogenizer comprises a high-pressure cavity, a homogenizing valve, a working cavity, a conveying system and a temperature control system, a slit is formed in the homogenizing valve, and the crude extracting solution located in the high-pressure cavity enters the working cavity through the slit of the homogenizing valve, the pressure intensity of the high-pressure cavity is 300-3000 bar, and the working temperature of the high-pressure homogenizer is 0-10 DEG C. The activation energy is reduced by controlling high-pressure and low-temperature environments, so that the original cell membrane structure is broken, the cell wall structure is destroyed, and phospholipid bimolecules are driven to aggregate and self-assemble to form the vesicles. For biological membranes derived from bacteria, fungi, plants and animals, the vesicles can be prepared by the preparation method disclosed by the invention, and the method has universality.

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 a prothallus proliferation culture medium and a rapid propagation method for rare and endangered plant cibotium barometz. The prothallus proliferation culture medium of cibotium barometz is used for accelerating the proliferation of gametophytes, and the prothallus maturation time is shortened; meanwhile, the invention provides a rapid propagation method of cibotium barometz, which comprises the following steps: carrying out artificial sterile culture on cibotium barometz spores, rapidly germinating in a culture bottle to form prothalli, culturing on a prothalli proliferation culture medium, inducing and transforming gametophytes in a specific matrix outside the bottle to form sporophytes by virtue of an incomplete tissue culture method, and transplanting. According to the method, the formation of prothallium is accelerated, the sporophyte conversion rate is increased, the reproduction rate is increased while the domestication and reproduction of sporophyte seedlings are accelerated, a large number of cibotium barometz seedlings which grow healthily can be obtained, the survival rate reaches 90%, the spore breeding period is obviously shortened, the operation requirement is not strict, and the method is suitable for large-scale popularization and application. The technical problems that the seedlings in the tissue culture bottle are easy to pollute, fragile and low in seedling rate are solved.

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 application finds that the heat resistance of a mutant material ski2dcl4 generated by a large amount of 22nt siRNA is obviously enhanced, and the heat resistance is mainly mediated by 22nt siRNA generated at an SMXL4 / 5 site; after thermal domestication, 22nt siRNA generated at the SMXL4 / 5 site can inhibit the expression of SMXL4 / 5 protein, so that the inhibition on HSFA2 is relieved; the specific targeting transcription inhibition factor SMXL4 / 5 can be used as a novel plant heat resistance method to improve the tolerance of plants to high temperature stress.

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 invention discloses aloe exosome-like nano-vesicles capable of delaying skin photoaging and an extraction method and application thereof, and belongs to the technical field of exosome-like nano-vesicle extraction. The extraction method of the aloe exosome-like nano-vesicles is designed and optimized, the yield of the aloe exosome-like nano-vesicles is obviously improved, and the extracted aloe exosome-like nano-vesicles can obviously delay skin photoaging and have an obvious improvement effect on skin fibroblast aging damage caused by ultraviolet radiation; a potential treatment means is provided for preventing or treating actinic keratosis caused by ultraviolet radiation and further possible squamous cell carcinoma.

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 belongs to the technical field of biological medicine, and relates to hydrogel for promoting wound healing and a preparation method and application thereof.The preparation method comprises the steps that firstly, coriander exosomes are extracted from coriander through a gradient centrifugation and sucrose density gradient method, then the coriander exosomes and a sodium alginate solution are mixed according to the mass-to-volume ratio of 0.5-2 mg: 1 mL, and a hydrogel solution is obtained; and mixing the exosome suspension with a calcium chloride solution according to a volume ratio of (0.5-2): 1, and carrying out crosslinking to obtain the coriander exosome-loaded hydrogel. The coriander exosome-loaded novel hydrogel provided by the invention has good viscoelasticity and biocompatibility, and can be applied to the surface of a skin wound as a dressing; meanwhile, the hydrogel has a coriander exosome slow-release effect, so that the exosome release time is prolonged, and the biological function is better played; by slowly releasing the coriander exosome, the hydrogel can relieve oxidative stress and inflammation of wounds and promote wound repair.

The invention relates to a preservation method and application of panax notoginseng germplasm resources, and relates to the technical field of panax notoginseng germplasm preservation. The preservation method comprises the following steps: (1) suspending pseudo-ginseng embryogenic calluses and a WPM culture medium, and screening and collecting to obtain pseudo-ginseng embryogenic cell clusters with the size of 0.45-1.5 mm; (2) transferring the panax notoginseng embryogenic cell cluster into a cryopreservation tube, and adding a loading solution for loading treatment to obtain a loaded embryogenic cell cluster; (3) carrying out vitrification treatment on the loaded panax notoginseng embryogenic cell cluster and a vitrification solution in a cryopreservation tube to obtain a vitrified panax notoginseng embryogenic cell cluster; (4) placing the cryopreservation tube filled with the vitrified pseudo-ginseng embryogenic cell cluster in liquid nitrogen for preservation; and after preservation, carrying out recovery culture on the pseudo-ginseng embryogenic cell cluster. According to the method, the survival rate reaches 90% after preservation, pseudo-ginseng plants can be rapidly differentiated, the differentiated pseudo-ginseng plants survive after being transplanted under natural conditions, the transplanting survival rate reaches 90%, and the pseudo-ginseng plants grow well in the later period.

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 provides a drug-loaded callicarpa nudiflora-derived extracellular vesicle as well as a preparation method and application thereof, and relates to the technical field of pharmaceutical preparations. The drug-loaded callicarpa nudiflora-derived extracellular vesicle has relatively high encapsulation efficiency and drug loading rate and good gastrointestinal tract stability and biocompatibility, can target a colitis disease part to reduce adverse reaction of a drug and improve the curative effect, solves the problem that the drug is insoluble in water, and can be used for preparing the drug-loaded callicarpa nudiflora-derived extracellular vesicle. The invention provides a novel drug delivery strategy for treating the ulcerative colitis, and is expected to become an effective strategy for clinically treating the ulcerative colitis, thereby achieving the effects of synergistically treating the ulcerative colitis and improving the effectiveness and safety of drug delivery.

The invention discloses a method for inducing sandalwood suspension cells to synthesize santalene. The method comprises the following steps: (1) performing induced callus culture on sandalwood explants; (2) carrying out suspension culture on the sandalwood callus to obtain a suspension cell line; and (3) adding cytokinin and auxin into the cell culture fluid of the suspension cell line, and performing induced culture to obtain the sandalwood suspension cell with high content of santalene. The method takes plant cell culture as a technical basis, and comprises the following steps: firstly, establishing a stable sandalwood suspension cell line; by adding cytokinin and auxin into the sandalwood cell culture fluid, the synthesis of the sandalene can be promoted, so that the content of the sandalene is increased. The result of the embodiment shows that according to the method provided by the invention, the biomass of the sandalwood suspension cells is improved by adding the cytokinin and the auxin. The product content in the sandalwood suspension cell line is about 3% and is obviously higher than that of a control group; gC-MS (gas chromatography-mass spectrometry) detection shows that the main components are alpha-santalene and beta-santalene.

The invention discloses a method for improving plant polyploidy induction rate and obtaining polyploidy based on leaf bud primordium cell cycle synchronization. The method comprises the following steps: carrying out pre-differentiation culture on an explant induction material; performing cell cycle synchronization excitation treatment on the leaf bud primordium cells obtained by the pre-differentiation treatment; and performing chromosome doubling treatment on the explant induction material subjected to the cell cycle synchronization excitation treatment. According to the method disclosed by the invention, an EdU staining labeling method and a flow cytometry are used for accurately judging the time for carrying out cell cycle synchronization excitation treatment on leaf bud primordium cells of an induction material, and carrying out synchronization excitation and doubling induction on the induction material in time; the dosage of a cell cycle inhibitor for synchronous excitation treatment is accurate, the treatment opportunity is accurate, the treatment duration is proper, and the polyploidy inductivity of the plant is remarkably improved; the obtained polyploidy plant is good in stability, and the method is high in repeatability.

The invention belongs to the technical field of biological agents, and particularly discloses application of lily extracellular vesicles in preparation of tumor treatment drugs. The preparation method of the lily extracellular vesicles comprises the following steps: S1, juicing lily bulb tissues, and filtering to obtain a lily bulb stock solution; s2, treating the lily bulb stock solution obtained in the step S1 to obtain a lily extracellular vesicle extracting solution; s3, concentrating and purifying the lily extracellular vesicle extracting solution obtained in the step S2 to obtain lily extracellular vesicles; wherein in the step S2, the treatment comprises centrifugation and filter membrane filtration. The lily extracellular vesicle provided by the invention can effectively promote polarization of M2-type macrophages to M1-type macrophages, shows remarkable immunoregulation ability, is good in biocompatibility, and has the potential of preparing anti-cancer drugs.

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 plant exosome extraction method and application thereof, and belongs to the technical field of exosome extraction. The extraction method of the plant exosome comprises the following steps: S1, mixing a plant sample with a buffer solution, performing juicing treatment, and filtering to obtain a crude extract; s2, carrying out continuous flow centrifugal treatment on the crude extract to obtain a refined extract; s3, carrying out concentration treatment on the refined extract to obtain a concentrated solution; and S4, carrying out microfluidic purification treatment on the concentrated solution to obtain the plant exosome. According to the method, the exosome of the plant sample is extracted by adopting a treatment method combining juicing treatment, continuous flow centrifugal treatment, concentration treatment and microfluidic purification treatment, so that large-volume, high-throughput, high-efficiency and low-cost exosome extraction and separation of the plant sample are realized; in addition, it is found for the first time that the panax notoginseng source exosome not only can effectively inhibit expression of inflammatory factors, but also has a good effect on psoriasis treatment.