630 results about "Vesicle" patented technology

The invention relates to the technical field of pharmaceutical preparations, and discloses a composition with liver protection and antioxidation effects and a preparation method thereof.The composition is prepared from galactosamine modified astragalus membranaceus exosome, schisandra chinensis nanocrystals, liquorice-chitosan nanoparticles, lactobacillus acidophilus freeze-dried powder, beta-(1, 3) / (1, 3, 4-tetramethyl-1, 3-pentanediol monoisobutyrate) sourced from saccharomyces cerevisiae, and beta-(1, 3, 4-tetramethyl-1, 3-pentanediol monoisobutyrate). And (6) a dextran, grape exosome-curcumin hybrid vesicle freeze-dried powder, a curcumin-mitochondrial targeting peptide compound, a resveratrol-hydroxypropyl-beta-cyclodextrin inclusion compound and reduced panthenol. According to the liver-protecting and anti-oxidation composition disclosed by the invention, multi-dimensional intervention on liver protection is realized through an elaborately designed ternary system. The system is composed of a traditional Chinese medicine extract, a microorganism-plant exosome combination and a high-bioavailability antioxidant, all the parts have a synergistic effect, and the liver protection effect is remarkably improved.

The invention discloses a therapeutic nano-vesicle preparation rich in mitochondrial functional protein, a preparation method of the therapeutic nano-vesicle preparation and application of the therapeutic nano-vesicle preparation in treatment of radioactive skin injury. The preparation method comprises the following steps: culturing, amplifying and cleaning umbilical cord mesenchymal stem cells, dispersing the umbilical cord mesenchymal stem cells in a suspension, mechanically extruding the umbilical cord mesenchymal stem cells into a crude solution through a multistage aperture polycarbonate membrane, and centrifuging, concentrating and purifying the crude solution to obtain the preparation. The preparation has the remarkable advantages that local administration can be realized, inflammation can be quickly inhibited, oxidative stress is reduced, and a repair pathway is activated, so that the preparation is suitable for treating acute radioactive skin injury; the preparation method is easy to standardize and scale, high in stability and convenient for emergency storage and delivery, and avoids the defects of cell therapy; a plurality of targets such as immune inflammation, oxidative stress and the like can be synergistically regulated, the biological effect of hUMSCs is simulated and partially replaced, and the limitation caused by the cell survival rate and the microenvironment dependency is avoided; the key function of hUMSCs is covered in function, and the clinical feasibility, safety and emergency suitability of the application level are higher.

The invention provides a method for detecting an external vesicle marker through high-flux nano plasma exciting light immune color development, and belongs to the technical field of human extracellular vesicles. After a serum sample is subjected to centrifugal treatment, the serum sample and a CD81 capture antibody substrate are incubated, and the particle size distribution of the vesicles is monitored in real time; a zwitterionic polymer modified gold nanoparticle LAM detection probe and a polyethylene glycol modified silver nanoparticle LprG detection probe are prepared to be specifically combined with a vesicle surface antigen, a chromogenic enhancement solution is adopted to induce a plasma resonance signal, and full-hole scanning imaging is carried out; and constructing a double-layer game optimization model to cooperatively optimize the detection sensitivity and the signal stability, carrying out weighted summation on normalized signals of the particle size subgroups to obtain comprehensive detection signal intensity, comparing the comprehensive detection signal intensity with a threshold value, and outputting a final judgment result. The technical problem that quantitative accuracy is affected by signal intensity deviation caused by vesicle particle size difference in outer vesicle marker detection is solved.

The invention discloses an engineered bionic nucleic acid nano-vesicle as well as a preparation method and application thereof, relates to the technical field of nano biomedicine, and aims at solving the problems that in-vivo targeting efficiency and immunogenicity of a traditional cationic lipid nano-carrier are limited due to deletion and cleavage obstacles of GSDMD expression in tumor cells. The technical key point of the invention is as follows: the engineered bionic nucleic acid nano-vesicle is provided and is prepared by wrapping a cationic lipid nucleic acid drug with an exosome derived from engineered macrophages; wherein the exosome from the engineered macrophage is the exosome from the macrophage with high expression of PD1, which is as shown in SEQ. ID. NO.1. The exosome from the engineered macrophage is the exosome from the macrophage with high expression of PD1; the lipid nucleic acid medicine is prepared by loading GSDMD-N mRNA (messenger Ribonucleic Acid) shown on the basis of SEQ.ID.NO.2 on a cationic liposome. The engineered bionic nucleic acid nano-vesicle is used for preparing an oral squamous cell carcinoma diagnostic kit and a therapeutic drug.

The invention provides a plug-and-play Mi3 protein nanocage fusion protein with a VNP6 tag and a preparation method and application of the plug-and-play Mi3 protein nanocage fusion protein, the fusion protein comprises the VNP6 tag, a SpyCatch003 component, a Mi3 protein nanocage and a functional protein sequence, and the VNP6 tag is located at the N end of the fusion protein. The VNP6 peptide sequence or the variant thereof is utilized to induce a vesicle or vesicle-like structure beneficial to protein folding and stabilization in escherichia coli, so that the correct folding rate and activity of recombinant protein are improved while the overall intracellular molecular crowding effect is relieved. The core idea of the invention lies in that the internal structure of cells is regulated at low temperature, and the internal microenvironment of an escherichia coli expression system is optimized, so that protein molecules under high expression load can obtain more reasonable spatial distribution, and the problems of misfolding and aggregation caused by molecular crowding are reduced.

The invention belongs to the field of biological medicines, and relates to a drug-loaded nano-vesicle as well as a preparation method and application thereof. The drug-loaded nano-vesicle comprises a vesicle core and a drug-loaded nano-vesicle, wherein the vesicle core comprises siRNA (small interfering Ribonucleic Acid) capable of specifically targeting and silencing an NR1D1 gene; the vesicle membrane is formed by fusing an erythrocyte membrane, a macrophage membrane, cardiolipin, cholesterol and lecithin. The drug-loaded nano-vesicle can specifically target macrophages in a sepsis immunosuppression stage, has an intracellular response release function, recovers BMAL1 and IGF2BP2-ATP6V1B2 / ATP6V0c axis functions by inhibiting NR1D1 expression, reconstructs a macrophage phagocytosis function and lysosome-dependent bacterium removal capability, and can be used for preparing a drug-loaded nano-vesicle with a specific targeting function. The survival rate of sepsis immunosuppression model animals is obviously improved; and the bacterial load is reduced. Compared with a traditional electroporation method, the preparation method disclosed by the invention has the advantage that the encapsulation efficiency of siRNA is remarkably improved.

The invention discloses a preparation method and application of a novel hybrid extracellular vesicle. According to the invention, vesicles derived from endothelial cells and neutrophil cells are combined with deferoxamine, and a biological mixed nano vesicle platform (DFO (HEVS)) is developed to solve the problem of diabetes wound healing. According to the dual-targeting system, DFO is accurately delivered to a wound part by utilizing CXCR4 mediated endothelial cell homing and beta2 integrin dependent inflammation tropism, DFO (at) HEVs activates and recovers vascular regeneration through HIF-1alpha / VEGF, ferroptosis is inhibited through Nrf2 / GPX4 signal transduction, macrophages are reprogrammed into a repair promoting M2 phenotype, and oxidative stress-inflammation-ferroptosis circulation is effectively broken.

The invention relates to the technical field of biomedical materials, in particular to engineered small extracellular vesicle hydrogel enriched with PD-L1 and Siglec-15 as well as a preparation method and application of the engineered small extracellular vesicle hydrogel. The engineering small extracellular vesicle hydrogel is enriched with PD-L1 and Siglec-15 at the same time, the engineering small extracellular vesicle hydrogel comprises a hydrogel base body and PDL1-Siglec15-sEVs loaded in the hydrogel base body, and the PDL1-Siglec15-sEVs are small extracellular vesicles with the PD-L1 and the Siglec-15 in an overexpression mode. Compared with natural small extracellular vesicles and a traditional wound surface treatment strategy, the engineered small extracellular vesicle hydrogel has the advantages that PD-L1 and Siglec-15 can be enriched in the vesicles, so that more accurate immune microenvironment regulation and control can be realized on the local part of a wound surface, excessive inflammatory response is moderately inhibited, phenotype transformation of macrophages to be beneficial to tissue regeneration is promoted, and the wound surface treatment effect is improved. The hydrogel is used as a carrier and can provide a moist healing environment and a three-dimensional scaffold structure, so that the residence time of the engineered small extracellular vesicles on the local wound surface is remarkably prolonged, and slow release is realized.

The invention discloses a symmetric nucleoside microneedle patch loaded with polymer vesicles as well as a preparation method and application of the symmetric nucleoside microneedle patch. The symmetric nucleoside microneedle patch loaded with the polymer vesicles comprises a guanosine-quadruplex hydrogel microneedle tip doped with drug-loading polymer vesicles, the raw material of the drug-loaded polymer vesicle comprises a polycaprolactone-polyglutamic acid copolymer; the guanosine-quadruplex hydrogel is prepared from the following raw materials: guanosine and KB (OH) 4. The microneedle patch is constructed on the basis of dynamic covalent bond and non-covalent interaction, has a sharp tip array capable of penetrating the epithelium and the lower layer of a mucous membrane, can efficiently deliver a drug to a canker lesion part by bypassing a mucous membrane barrier, can be controllably broken under a high ROS condition of an inflammation microenvironment, has the characteristics of high drug delivery efficiency and high accuracy, and can be used for preparing a medicine for treating the canker lesion part. The effect of effectively improving chemotherapy-induced oral ulceration can be achieved by reducing inflammatory response, relieving pain feeling and inhibiting pain sensitization.

The invention discloses a plant cell vesicle efficient extraction method based on the synergistic effect of three-dimensional gradient micro vortex and sound wave fluidization and application of the plant cell vesicle efficient extraction method. According to the plant cell vesicle extraction method provided by the invention, high-yield, high-purity and high-activity extraction of plant cell vesicles is realized through an active enzymolysis-acoustic cracking-three-dimensional gradient separation full-chain process, and the plant cell vesicle extraction method can be used for large-scale production. The method overcomes the defects of low yield and purity, chemical pollution, low extraction efficiency, uncontrollable vesicle size, easily damaged structure, long time and the like of the traditional extraction method, and can be used for producing and preparing more cosmetics and medicines containing plant cell vesicles on a large scale. The method systematically solves the global problem of extraction and application of the plant vesicles through multidisciplinary cross innovation (acoustics, fluid mechanics and molecular bionics), is high in extraction efficiency, good in activity effect and comprehensive and advanced in technical parameters, and has important significance on extraction and development of the plant cell vesicles.

The invention discloses a preparation method and application of a dual-targeting carrier-free delivery system for enhancing anti-tumor immunity of NKT cells, Lys-alpha-GalCer is used as a raw material, and two molecules of LGC are connected through a disulfide bond to synthesize an amphiphilic LGC-SS-LGC micromolecule prodrug; the LGC-SS-LGC micromolecule prodrug is prepared into vesicles internally loaded with an NKT cell antigen presenting molecule CD1d and externally modified with polysarcosine through electrostatic adsorption through a film dispersion-probe ultrasonic method, finally a CD1d / LGC-SS-LGC / PSar dual-targeting carrier-free delivery system is formed, and the dual-targeting carrier-free delivery system can be used for liver cancer treatment after intravenous administration. In addition, the compound can be used for preparing a preparation for enhancing NKT cell Th1 type immune response. The dual-targeting carrier-free delivery system prepared by the invention can directly or indirectly enhance the anti-tumor immune effect of the NKT cells, and has a dual-synergistic effect.

The invention discloses an extraction method of safflower leaf exosome-like nano-vesicles and application of the safflower leaf exosome-like nano-vesicles in preparation of drugs for preventing and treating adriamycin-induced cardiotoxicity, fresh safflower leaves are taken and cleaned, a PBS solution is added for crushing and filtering, filtrate is collected and continuously centrifuged, supernate is taken and subjected to ultracentrifugation, precipitates are collected after centrifugation, and the safflower leaf exosome-like nano-vesicles are obtained; and filtering and sterilizing by using a filter membrane to obtain the safflower leaf exosome-like nano-vesicles. The carthamus tinctorius leaf exosome-like nano-vesicles can significantly improve the heart function of doxorubicin-induced cardiotoxic mice; myocardial injury induced by doxorubicin can be repaired, and myocardial fibrosis induced by doxorubicin can be inhibited; the content of a myocardial injury marker lactic dehydrogenase in body serum can be reduced, a new treatment strategy is provided for preventing and treating adriamycin-induced cardiotoxic diseases, and the application prospect is good.

The invention discloses an application of exosome nano-vesicles derived from artemisia annua cells and peroxidase A0A2U1N9S9 contained in the exosome nano-vesicles in preparation of anti-colorectal cancer drugs. According to the present invention, the anti-CRC activity analysis results show that the artemisia annua cell-derived exosome nano-vesicles can inhibit the growth of DLD-1 and HCT116 cell strains, and the active substance base of ACDENVs for inhibiting the growth of DLD-1 and HCT116 cell strains, namely peroxidase A0A2U1N9S9, is innovatively revealed for the first time based on the difference analysis of the proteomics technology; and a new research thought and a theoretical basis are provided for the research of anti-colorectal cancer drugs.

The present disclosure provides ghost nanovesicles (gNVs) that are deficient in cytosolic components. Methods of making such vesicles and therapeutic uses of such vesicles are also provided. The gNVs may be used in preventing or treating conditions that may benefit from administration of the gNVs. Such conditions include conditions that involve inflammation.

The invention discloses a method suitable for extracting extracellular small vesicles in fresh water pearl shell tissue waste liquid, which comprises the following operation steps: (1) solid-liquid separation: slaughtering pearl shells, and filling slaughtering waste liquid into a waste liquid crude extraction device; (2) waste liquid crude extraction: performing multi-stage gradient filtration and gravity settling on the slaughtering waste liquid by using a waste liquid crude extraction device to obtain clarified waste liquid; (3) removing impurities: removing impurities for the first time and removing impurities for the second time; (4) concentration of the extracellular vesicles: concentrating the extracellular vesicles by using an ultra-speed centrifuge to obtain a high-concentration extracellular vesicle solution; and (5) preservation of extracellular vesicles: according to the final volume of the high-concentration extracellular vesicle solution obtained in the step (4), carrying out sample standardization through protein concentration, sub-packaging the sample, and carrying out long-term preservation in a low-temperature environment. The particle sizes of the extracted vesicles are concentrated, high-purity and complete extracellular small vesicles can be obtained, and the method is suitable for biomedical research and product development.

The invention discloses a method for annotating a vesicle cell subset based on monovesicle membrane proteomics, and belongs to the technical field of vesicle cell annotation. The method comprises the following steps: 1, carrying out exosome collection on a plasma sample to obtain a sample; 2, coding and labeling the antibody probe, and adding a protein tag and a labeled DNA sequence into the antibody probe to obtain a labeled antibody probe; 3, preparing a corresponding combined product; 4, adding a labeled antibody probe into the sample to obtain an exosome compound; step 5, capturing an exosome conjugate through CTB; 6, adding the binding product into the exosome conjugate to generate a sequence; 7, constructing a sequence library and sequencing to obtain a sequencing sequence; 8, removing a sequencing sequence with low expression quantity to obtain an exosome expression data matrix, and standardizing the exosome expression data matrix; 9, carrying out clustering analysis to find out an exosome core subgroup; and step 10, determining the source of the exosome core subgroup, and annotating the single vesicle subgroup.

The invention discloses a vesicle transportation path prediction system based on artificial intelligence, and relates to the technical field of vesicle transportation, which comprises the following steps of: accurately extracting a complete time sequence movement track of a single vesicle before and after disturbance by virtue of fluorescence living cell imaging under a disturbance experiment condition in combination with a target detection model YOLOv8 and a depth correlation tracking algorithm DeepSORT; the problem that perturbation conditions are insufficiently perceived is solved, through a multi-modal data matrix, the model has the time sequence perceiving capacity for changes of a local space structure where vesicles are located, so that the sensitivity and modeling capacity of the model for track variation in a complex environment are remarkably improved, deep fusion of multi-modal features is achieved by building a double-branch fusion model, and the accuracy of the model is improved. The problem that an existing prediction model cannot actively recognize and correct prediction deviation is effectively solved, and the stability and the actual usability of the system are improved.

The invention is applicable to the technical field of biology, and provides a method for rapidly extracting plant rhizome extracellular vesicles from monosaccharide and compound enzyme. According to the method, the plant extracellular vesicles are extracted by an enzymolysis-monosaccharide solution extraction coupling method for the first time, and the defect that an extraction method is single is overcome. According to the method, the conditions are mild, the primary property, the integrity and the biological activity of the vesicle structure are kept, the extraction purity is high, and the particle size distribution is uniform and concentrated in the range of 20-300 nm. The total protein extraction amounts of glucose, galactose and fructose solutions are respectively increased by 36.09% + / -1.18%, 37.65% + / -2.83% and 36.83% + / -2.59% compared with those of PBS (Phosphate Buffer Solution) extraction treatment. In addition, the'single ultracentrifugation 'purification process overcomes the problem of time consumption of traditional gradient centrifugation, and the time is shortened by more than 3h compared with that of a traditional extraction method. The method breaks through the technical bottleneck of large-scale production of the plant extracellular vesicles, and has great application and popularization values.

The invention discloses a grifola frondosa-derived exosome-like nano-vesicle as well as an extraction method and application thereof. The extraction method comprises the following steps: cutting pilei and stipes of grifola frondosa, and grinding with a proper amount of cold PBS to prepare grifola frondosa bacterial juice; the grifola frondosa juice is centrifuged at 500 g-1, 000 g, sediment is discarded, the obtained supernatant is centrifuged at 2,000 g-3,000 g, sediment is discarded, the obtained supernatant is centrifuged at 10,000 g-12,000 g, sediment is discarded, the obtained supernatant is filtered, and the pretreated grifola frondosa juice is obtained; and centrifuging the pretreated grifola frondosa juice at 30,000-100,000 g for 30 minutes to 4 hours, and resuspending by using PBS (Phosphate Buffer Solution), so as to obtain the grifola frondosa-derived exosome-like nano-vesicles. The invention develops a new thought for the mechanism of cancer inhibition, chemotherapy effect enhancement and flora regulation of the grifola frondosa, and lays a foundation for application of the grifola frondosa in development of products such as related foods, functional foods, health foods, medicines and gel-loaded medicines.

The present disclosure relates to a CCL20 promoter derived from a regulatory region of the CCL20 gene, which stimulates gene expression in response to inflammation. These CCL20 promoters, which can be stimulated by endogenous or exogenous cytokines, can be used to control the timing of expression of heterologous genes. The disclosure also relates to expression cassettes or vectors comprising a CCL20 promoter of the disclosure operably linked to a nucleic acid sequence encoding, for example, a polypeptide of interest; and delivery systems (e.g., viral particles, lipid vesicles or nanoparticles) or cells comprising such expression cassettes or vectors. The disclosure also relates to the use of the promoter, expression cassette, vector, delivery system or cell in the treatment of inflammation-related diseases or for recombinant gene expression.

The invention discloses a preparation method of a plant source nano vesicle and the plant source nano vesicle. According to the preparation method disclosed by the invention, through extensive and deep research and a large amount of screening optimization, the coix seed-derived nano vesicles (SCDNVs) and the gastrodia elata-derived nano vesicles (RGDNVs) which are high in concentration and high in quality are successfully extracted by using the preparation method disclosed by the invention; meanwhile, the invention further researches the anti-tumor effect and the anti-oxidation effect of the prepared plant source nano vesicle. Through screening of conditions of the preparation method, an ultrasonic combined centrifugal method (further comprising screening of ultrasonic conditions, such as ultrasonic power / ultrasonic time and the like) adopted in the invention can overcome the defects that the membrane structure of the vesicle is damaged and the form of the vesicle is not complete enough in the traditional method; therefore, the plant extracellular vesicles which are small in loss, high in concentration, good in stability, more uniform in vesicle particle size, small in particle size span, high in yield and high in quality are obtained.

The invention discloses a PDRN-loaded umbilical cord mesenchymal stem cell source nano-vesicle and a preparation method thereof, and belongs to the technical field of biological medicines. The preparation method provided by the invention comprises the following steps: mixing an umbilical cord mesenchymal stem cell suspension with a PDRN solution, and carrying out freeze thawing treatment; incubating the mixed solution subjected to freeze thawing treatment; extruding the incubated mixed solution for multiple times by using a filter membrane; and collecting the extruded product, and separating and purifying to obtain the PDRN-loaded umbilical cord mesenchymal stem cell source nano-vesicle. According to the preparation method disclosed by the invention, the encapsulation efficiency of PDRN is remarkably improved, meanwhile, secondary damage of a traditional post-loading technology to a preformed vesicle structure is avoided, the membrane integrity and biological activity of a final product are better maintained, the yield is high, and the preparation process is rapid; the introduction of exogenous impurities is avoided from the source, and the high purity and biological safety of the obtained nano-vesicle component are ensured.

The invention provides a preparation method of a substrate of an external vesicle functional marker rapid detection kit, and belongs to the technical field of external vesicle functional marker detection.The preparation method comprises the steps that a high-purity coating working solution is prepared, and precise spraying is achieved through a piezoelectric type ink jet system; substrate functional modification is completed through confining liquid treatment and labeled antibody working solution preparation, a double-layer game model containing detection sensitivity optimization and process stability optimization is established, and an optimal parameter combination is obtained through game iterative calculation by combining the synergistic effect of a quality evaluation function and a stability control function. A characteristic parameter database is established to store historical production data and quality detection results, a multi-parameter joint evaluation method is adopted to calculate a comprehensive score and carry out unified product grading, and a quality control system and a standardized parameter optimization mechanism are driven through full-process data. The technical problem that the quality consistency difference is large in the batch production process is solved.

The invention relates to an adipose-derived stem cell and exosome composition for mucosal inflammation and an application of the adipose-derived stem cell and exosome composition. The composition is composed of a hybrid nano-vesicle and a pulsatilla chinensis exosome compound in a mass ratio of 1: 1. The hybrid nanovesicles are prepared by fusing adipose-derived stem cell exosomes and lipidosome and modifying honey polysaccharide, and the pulsatilla chinensis exosome compound is loaded with interleukin 23 peptidomimetic, polyethylene glycol, black scale quantum dots and camptothecin. The composition has the functions of efficient anti-inflammation, immunoregulation, precise targeting and tissue repair, and meanwhile, the curative effect is enhanced through photo-thermal synergistic treatment. The preparation method comprises the steps of exosome extraction by a density gradient centrifugation method, liposome fusion and surface modification. The composition is suitable for treating mucosal inflammation such as inflammatory bowel disease and oral mucositis, has the advantages of high stability, good biocompatibility and the like, and provides a safe and effective novel treatment strategy for clinic.

The invention belongs to the technical field of bionic cilium sensors, and particularly discloses a magnetic bionic cilium and vesicle complex and a preparation method and application thereof.The preparation method comprises the following steps that S1, a cilium array male mold is prepared; then carrying out oxygen plasma treatment, silanization and curing stabilization; s3, injecting the magnetic composite precursor into a polydimethylsiloxane female die, applying pressure, and striking off; and S4, coating the mold surface with an Ecoflex precursor, performing thermocuring forming, performing magnetization treatment, and performing stripping to obtain the magnetic bionic cilia. According to the magnetic bionic cilia, the vesicle complex and the preparation method and application of the magnetic bionic cilia, the vesicle complex and the preparation method and application of the magnetic bionic cilia, high-magnetic signal output is achieved while the deformability of the cilia is guaranteed, the magnetic bionic cilia is used for non-contact real-time hemodynamic monitoring, meanwhile, specific response release of the vesicle complex is achieved, and a new thought is provided for design of responsive vascular implants.

The invention relates to a repairing composition for reducing senescence cell cumulant and a preparation method and application thereof, and the repairing composition comprises the following components in parts by weight: 0.01-7.0 parts of probiotic pellets PDRN, 0.01-3.0 parts of microspheroidized PDRN, 0.01-10.0 parts of a ginseng root extract, 0.01-10.0 parts of red ginseng fermented vesicles, a thickening agent, functional components and water. Under a formula system of a thickening agent and functional components, the probiotic pellets PDRN, the microspheroidized PDRN, the ginseng root extract and the red ginseng fermented vesicles are added, and all the components have a synergistic effect, so that the composition can penetrate into skin to achieve the effect of accurately repairing mitochondrial dysfunction, can directly remove senescent cells, reduces the senescent cell cumulant in a body in multiple ways, and has a good effect on repairing mitochondrial dysfunction. Therefore, the prepared skin care composition can effectively delay skin aging.

The invention discloses an engineered artificial vesicle and application of the engineered artificial vesicle in multiple in-situ detection of urine exosome miRNA, and belongs to the field of biosensors. The method comprises the following steps: constructing planar framework nucleic acid simultaneously modified with cholesterol and an aptamer through annealing reaction; a molecular beacon with a fluorophore is designed according to a target gene sequence, and the molecular beacon and a double-strand specific nuclease (DSN) system are jointly encapsulated in an artificial vesicle; the engineered artificial vesicles and a sample to be detected are incubated, the fluorescence modified molecular beacons can specifically recognize target genes, cyclic cutting and signal amplification are achieved under DSN mediation, and the broken molecular beacons release fluorescence signals; all fluorescence signals in the vesicles are collected through a fluorescence imaging technology, and a characteristic fluorescence spectrum of the to-be-detected sample is obtained. According to the present invention, the miRNA heterogeneity analysis at the single vesicle level and the precise multi-target detection of bladder cancer and other diseases can be achieved, and the problems of low throughput, target number limitation and the like of the existing vesicle in-situ detection are effectively overcome.

The invention discloses a single vesicle particle electrochemical signal identification and classification method and identification system, and the method comprises the steps: S1, preparing a carbon fiber ultramicro electrode, applying a voltage in a patch clamp system constant potential mode, and collecting picoampere-level current response generated by oxidation of a single vesicle content on the surface of the electrode; s2, carrying out smooth filtering on the collected original current signal, and realizing characteristic peak identification and multi-dimensional physical / statistical characteristic extraction by using a multi-parameter peak identification method based on a threshold value; s3, carrying out preprocessing operations including missing value processing, standardization, principal component analysis dimension reduction and category equalization on the extracted peak value characteristic data; and S4, on the basis of the preprocessed feature data, constructing and training at least one machine learning model for automatically identifying the particle size category of the vesicle particles. The method combines the advantages of single-vesicle electrochemical high-sensitivity measurement and intelligent algorithm classification, and has the advantages of high recognition efficiency, good repeatability, wide applicability and the like.

The invention is applicable to the technical field of biology, and provides a method for rapidly extracting plant rhizome extracellular vesicles by disaccharide and compound enzyme. According to the method, an enzymolysis-disaccharide solution extraction coupling method is created for the first time to extract the plant extracellular vesicles, and the defect that an extraction method is single is overcome. According to the method, the conditions are mild, the primary property, the integrity and the biological activity of the vesicle structure are kept, the extraction purity is high, and the particle size distribution is uniform and concentrated in the range of 40-300 nm. Compared with PBS (Phosphate Buffer Solution) extraction treatment, the total protein extraction amounts of lactose, cane sugar and maltose solutions are respectively increased by 20.04% + / -4.50%, 35.00% + / -5.74% and 36.45% + / -2.68%. In addition, the'single ultracentrifugation 'purification process overcomes the problem of time consumption of traditional gradient centrifugation, and the time is shortened by more than 3h compared with that of a traditional extraction method. The method breaks through the technical bottleneck of large-scale production of the plant extracellular vesicles, and has great application and popularization values.

The invention discloses a method for separating and detecting new intestinal bacterial exovesicles, which comprises the following steps: (1) carrying out metabolism labeling on microorganisms to be researched by an azido-modified hydrochloric acid D-type amino acid metabolism marker, so that the new intestinal bacterial exovesicles carry azido groups; (2) separating and extracting the bacterial exovesicles from a sample containing the bacterial exovesicles; (3) adding a capture connector capable of reacting with an azide group into the extracted bacterial outer vesicles; and (4) separating, capturing and detecting the bacterial outer vesicles obtained in the step (3) by adopting a micro-fluidic chip, wherein the obtained bacterial outer vesicles are newborn bacterial outer vesicles. The method disclosed by the invention can selectively capture the newborn bacterial exovesicles from the sample under the condition of not influencing surface protein expression of the bacterial exovesicles so as to carry out downstream analysis on the newborn bacterial exovesicles of the intestinal flora.