24 results about "Hirudin" patented technology

This invention relates to the field of health products or food technology, and more specifically to a composition and its application in removing dampness and controlling blood sugar, balancing blood pressure, and regulating blood lipids. The invention provides a composition for removing dampness and controlling blood sugar, balancing blood pressure, and regulating blood lipids, comprising Poria cocos extract, Astragalus membranaceus extract, hirudin, and at least one or more of Eucommia ulmoides extract, Gynostemma pentaphyllum extract, and Morus alba twig extract. The composition provided by this invention utilizes extracts from at least four precious Chinese medicinal herbs to exert a synergistic effect, jointly achieving the effects of lowering blood pressure, removing dampness and lowering blood sugar, and lowering blood lipids.

The invention discloses a hirudin escherichia coli engineering bacterium with controllable production generations as well as a construction method and application thereof, and relates to the technical field of biological medicines. The invention discloses a construction method of hirudin escherichia coli engineering bacteria with controllable production generations, which comprises the following steps: constructing a hokC protein gene as shown in the sequence of SEQ ID NO.1 and a constitutive promoter trxA as shown in the sequence of SEQ ID NO.2 into a recombinant expression vector pET28a (+)-ptrxA-hokC, and then transforming the recombinant expression vector into a hirudin synthetic strain E. coli Hirudin HV1-3 to obtain the hirudin escherichia coli engineering bacteria with controllable production generations. Through multiple rounds of culture, the hirudin yield of the engineering strain is reduced to 0 in the tenth round. The method is suitable for various hirudin types, and has the characteristics of reliable operation and high safety.

The invention relates to an exosome with high permeability and application of the exosome, and the surface of the exosome is connected with dioleoyl phosphatidylserine, 1, 2-dioleoyl-sn-glycerol-3-phosphorylethanolamine and hirudin. The exosome constructed by the invention can improve the permeability of the skin, can carry active components to directly reach the skin epidermis and corium layer, is accumulated on the epidermis and corium layer, continuously provides nutrition for the skin, and fully exerts the skin care and beautifying effects.

The invention belongs to the technical field of nano preparations, and particularly relates to a hirudin nano drug for relieving chronic asthma and a preparation method thereof.The hirudin nano drug is prepared from, by weight, 0.5-1 part of hirudin prodrug, 10-15 parts of stabilizer, 40-80 parts of freeze-drying protective additive and 1.5 parts of histidine; the hirudin prodrug molecule is composed of hirudin, MMP-9 responsive peptide, ROS responsive thioether bond and hydrophobic hexadecanoic acid, can spontaneously assemble to form a nano-sized prodrug under hydrophilic and hydrophobic effects, has stability in systemic circulation and is targeted and enriched at an asthma airway inflammation part; the hirudin with the thrombin activity inhibition effect is responsively released at the focus, so that the effect and the purpose of slowly treating the asthma are achieved.

The present application belongs to the technical field of biological medicine separation and purification, and particularly relates to a leech-derived hirudin separation and purification method based on thrombin affinity magnetic microspheres and application thereof. The present application uses self-prepared leaching liquor to coarsely extract hirudin from leech powder, then selects carboxyl magnetic microspheres to prepare thrombin affinity magnetic microspheres, and further separates and purifies by using the thrombin affinity magnetic microspheres, so as to finally obtain natural hirudin with an activity of 17000 ATU / g or more. The self-prepared leaching liquor can destroy the cell wall of leech, so that the hirudin in the leech is fully dissolved, thereby providing higher efficiency for the coarse extraction of hirudin in leech powder. The prepared thrombin affinity magnetic microspheres can fully utilize the sample enrichment effect thereof, and compared with agarose microspheres and polymer microspheres, can separate and purify hirudin with higher activity and purity, has a long storage period and stable quality, can provide safe and high-quality raw materials for health food, medicine or cosmetics, and is beneficial to industrialized production and application.

The invention relates to the technical field of traditional Chinese medicinal material processing, in particular to a leech fresh-cut processing device and method. Comprising the steps that a conveying mechanism is installed on a workbench so as to transversely fix and convey leeches, and an intelligent air control guiding-in mechanism and a fresh cutting mechanism conduct section limiting cutting on the leeches in a channel; and the fresh cutting mechanism is connected with the controller, identifies the position of the leech based on the laser signal as a starting signal, controls the conveying mechanism to stop in a delayed manner so as to adjust the length of the feed opening of the leech, and is linked to cut the leech in a node limiting manner. Impurities are removed from the source, the relative content of hirudin and other antithrombotic polypeptides is remarkably increased, the color and smell of a finished product are improved, the risk of pathogenic bacteria is reduced, efficient and standardized production is achieved through automatic equipment, and the problems of component dilution, quality degradation, low efficiency and the like existing in a traditional processing mode are solved.

The invention discloses a natural hirudin extraction method and application, and relates to the technical field of natural hirudin extraction.The method is characterized by comprising the steps that a tissue structure capable of being subjected to salivary gland single cell combined gland culture is selected; washing away bloodstain in the tissue structure by using PBS, then washing for three times by using the antibacterial liquid, and finally washing away the residual antibacterial liquid by using PBS; inoculating the tissue structure to a cell culture plate, placing the cell culture plate in a 28 DEG C incubator for inverted culture for 2 hours, then adding a DMEM / F-12 complete medium containing a growth additive and not containing serum, and placing the cell culture plate in the 28 DEG C incubator; when the tissue structure generates a large number of secretory vesicles, collecting the culture solution, adding a new culture solution, taking the collected culture solution as a natural hirudin crude product, and separating and purifying the natural hirudin crude product to obtain the natural hirudin. The method can achieve the effects of long-term survival in vitro and generation of a large amount of secretions.

The invention provides a multi-target synergistic cardiovascular protection beverage which comprises functional components and auxiliary components, and the functional components comprise nattokinase, hirudin, squalene, EPA, DHA and phytosterol; the preparation method of the multi-target synergistic cardiovascular protection beverage comprises the following steps: S1, firstly, washing the functional components with deionized water respectively to remove surface impurities, dust and microorganisms for subsequent processing; compared with the prior art, the raw material drying device has the following beneficial effects that in the drying process, flowing of hot air can take away moisture on the surfaces of raw materials, the humidity gradient is formed, the drying process is more uniform, and the phenomenon that the raw materials are locally overheated or dried unevenly is avoided; the method disclosed by the invention has the following beneficial effects that the raw materials are washed by deionized water and are dried by vacuum or low-temperature hot air, so that impurities and microorganisms can be thoroughly removed, and the loss of heat-sensitive functional components can be reduced.

The invention provides a hirudin penetration-promoting herbal compound body fat health-care liquid as well as a preparation method and application thereof, and the hirudin penetration-promoting herbal compound body fat health-care liquid comprises the following raw materials in percentage by weight: 0.3-0.6% of a leech extract, 0.3-0.6% of a Chinese herbal medicine extract and the balance of water. 5-10% of a herbal compound extract; 0.3 to 0.5 percent of methyl salicylate; 0.2 to 0.4 percent of ginger root oil; 8-12% of ethanol; and the balance of water. Wherein the herbal compound extract comprises an aloe vera extract, an astragalus membraneus extract, a ginseng extract and a panax notoginseng leaf and stem extract. According to the invention, a three-in-one technical system of efficient permeation promotion, multi-element conditioning, stability and mildness is constructed, the technical bottlenecks of low transdermal efficiency, single effect and insufficient stability of a traditional external herbal product are broken through, and the synergistic gain of body fat conditioning and skin repair is realized.

The invention provides a high-activity hirudin freeze-drying preparation as well as a preparation method and application thereof, and the preparation method comprises the following steps: (1) introducing a hirudin gene to a chassis cell to construct a recombinant engineering strain; (2) inoculating the engineering strain into a prepared culture medium, carrying out fermentation culture at a set temperature, after the thallus density reaches a target density, switching to methanol induced expression, and after continuous induction is carried out for a preset time, extracting a fermentation liquid; (3) performing ultrafiltration concentration and chromatographic purification on the obtained fermentation liquor to obtain a purified hirudin solution; and (4) adding a stabilizer into the obtained hirudin solution, pre-freezing to completely cure the hirudin solution, and drying to obtain the target hirudin freeze-dried preparation. According to the invention, the high-activity, high-purity, high-stability and low-cost industrial production of the hirudin freeze-drying preparation is realized through collaborative optimization of genetic engineering strain construction, high-density fermentation, multi-step purification and freeze-drying processes.

The invention relates to the technical field of biological medicines, and particularly discloses a hirudin-containing composition and application thereof in preparation of a product with a scalp absorption improving effect. The hirudin-containing composition is prepared from hirudin and a traditional Chinese medicine extract, the traditional Chinese medicine extract is prepared from the following raw materials: herba ecliptae, cacumen biotae and rhizoma drynariae. Researches show that the composition has the effect of promoting hair follicle cell proliferation; in addition, the absorption on the scalp can be improved; the important application value is realized.

The invention belongs to the technical field of traditional Chinese medicine pharmacy, and particularly relates to poecilobdella manillensis freeze-dried powder as well as a preparation method and application thereof. The preparation method of the poecilobdella manillensis freeze-dried powder comprises the following steps: (1) taking living poecilobdella manillensis or freezing and melting the living poecilobdella manillensis, and homogenizing to obtain a poecilobdella manillensis sample solution; (2) carrying out water extraction on the poecilobdella manillensis sample liquid; (3) performing fat extraction on the poecilobdella manillensis precipitate subjected to water extraction; and (4) combining the two extracts to prepare freeze-dried powder. The hirudin content of the poecilobdella manillensis freeze-dried powder prepared by combining a water extraction method with a fat extraction method is remarkably higher than that of a commercially available poecilobdella manillensis product, and the poecilobdella manillensis freeze-dried powder has a remarkable effect on treating thrombotic diseases and other vascular diseases.

The utility model belongs to the technical field of hirudin extraction, and particularly relates to an extraction device for hirudin, which comprises an extraction box. Living leeches are placed in a filter box, then a top cover is fixed, a driving motor drives an eccentric wheel to rotate, the eccentric wheel extrudes a ball when rotating for the first half circle, the ball drives an L-shaped plate to move rightwards, the L-shaped plate drives a connecting plate to slide on a guide rod and compress a first spring, and the connecting plate drives the filter box to move rightwards. When the eccentric wheel rotates to the rear half circle, extrusion on the ball is gradually released, the first spring is reset, the first spring drives the filter box to move leftwards through the connecting plate, the filter box shakes left and right in a reciprocating mode, then the leeches are stimulated to automatically spit saliva, and then the saliva of the leeches is flushed to the bottom of the extraction box through the filter box by spraying water through a spray head; when hirudin is separated out in water, shaking of the filtering box is achieved, the leeches are stimulated to secrete saliva more actively, and therefore the extraction period is shortened.

The application provides an extraction method of hirudin, an oral preparation containing the hirudin and a preparation method of the oral preparation, and belongs to the technical fields of hirudo processing and traditional Chinese medicine. The extraction method of the hirudin comprises the following steps: S1, soaking fresh hirudo in a freeze-drying protective agent, wiping the surface after taking out, pre-freezing at-40 to-50 DEG C for 1 to 2 hours, and then freeze-drying at-20 to-30 DEG C for 4 to 6 hours to obtain freeze-dried hirudo; S2, crushing the freeze-dried hirudo, adding water, adding enzymes and lecithin, and enzymolysis at 35 to 40 DEG C and pH=7.5 to 8.5 for 4 to 6 hours, and then cooling to room temperature to obtain an enzymolysis product; and S3, sterilizing, ultrafiltering and drying to obtain the hirudin. The hirudin obtained by the method has high activity and high recovery rate.

The invention provides a PMP hollow fiber membrane with an antithrombotic function and a preparation method of the PMP hollow fiber membrane. According to the method, a decahydronaphthalene / xylene low-temperature solvent system is constructed, PMP is uniformly dissolved under the condition that the temperature does not exceed 90 DEG C, and a thermal protective agent is adopted, so that the biological activity of hirudin in the processing process is effectively maintained; furthermore, oxidized chitosan is used as a controlled release carrier, and high load and slow release regulation of hirudin are realized through directional covalent coupling grafting of the zwitterionic polymer; finally, the physical barrier of the zwitterionic polymer and the direct anticoagulation effect of the hirudin are combined to construct a synergistic antithrombotic mechanism. On the premise of not depending on heparin, the obtained hollow fiber membrane has the comprehensive advantages that the blood coagulation time is remarkably prolonged, hirudin is released stably and durably, and HIT risks are avoided, is suitable for clinical scenes such as blood purification and in-vitro oxygenation, and has important clinical application value.

This invention discloses an antithrombotic medical coating, its preparation method, and its application. The antithrombotic medical coating is prepared by mixing an aqueous resin, hirudin-loaded PLGA / CS microspheres, a dispersant, a wetting agent, a pH adjuster, a film-forming aid, a foaming agent, and deionized water. The hirudin-loaded PLGA / CS microspheres are drug-loaded microspheres with a double-shell core-shell structure, consisting of hirudin as the core, PLGA as the inner shell, and CS as the outer shell. The antithrombotic medical coating can be used to prepare an antithrombotic coating containing microneedles. The prepared microneedle-loaded coating not only has the advantages of active permeation promotion, intelligent sustained release, and comfortable breathability, but also achieves non-invasive, long-lasting, convenient, and comfortable antithrombotic prevention functions, significantly improving adherence to antithrombotic treatment. Furthermore, it is adaptable to various flexible substrates, easily manufactured into various wearable antithrombotic products, and offers flexible application options. It also solves the common problems of brittle fracture and low yield in the preparation of microneedle patches.

The application provides a kind of palace pill for clearing palace and removing blood stasis and a preparation method thereof, which is prepared from the following raw materials in parts by weight: 25-30 parts of wine red peony root, 15-20 parts of vinegar-fried aconite, 15-20 parts of bran-fried broad wood, 15-20 parts of fried five spirit resins, 15-20 parts of vinegar-fried corydalis, 10-15 parts of hirudin, 5-10 parts of earthworm protein peptide powder and 5-10 parts of oyster peptide powder. The preparation steps of the oyster peptide powder are as follows: fresh oysters are selected, washed with running water, cleaned of internal organs and mucus, soaked in a citric acid solution, ultrasonically broken, enzymatically hydrolyzed, deodorized, purified, filtered, concentrated and dried to obtain oyster peptide powder. The palace pill has a scientific formula, a synergistic effect of various components, the efficacy of clearing palace and removing blood stasis, heat-clearing and detoxifying, carbuncle-dissolving and pus-discharging, blood stasis-removing and pain-stopping, is convenient to carry and take, and has clinical drug value.

This invention discloses a small molecule peptide derived from *Hirudo medicinalis*, with the amino acid sequence Leu-Leu-Phe-Arg. The peptide was obtained by enzymatically hydrolyzing the waste protein residue after purifying hirudin from *Hirudo medicinalis*, combined with tyrosinase-functionalized magnetic bead affinity fishing technology and mass spectrometry identification. In vitro kinetics and molecular dynamics simulations confirmed that the peptide LLFR significantly prolongs the lag time of monophenolase reactions. It penetrates the catalytic pocket of tyrosinase through an "induced fit" mechanism, forming a stable dead-end complex with the enzyme and inducing the unfolding of the enzyme's tertiary structure. Cell experiments confirmed that LLFR significantly inhibits tyrosinase activity and melanin synthesis in melanoma cells without significant cytotoxicity. This peptide exhibits high safety and good water solubility, providing a new approach for the high-value utilization of industrial waste from *Hirudo medicinalis*, and is suitable for the industrial production of whitening cosmetics and functional products.

The invention is applicable to the technical field of hirudin extraction, and provides a method for extracting and separating hirudin series products from living leeches, which comprises the following steps: feeding adult poecilobdella manillensis subjected to fasting starvation treatment with an inducer, adding a small molecule emetic to promote emesis, and collecting culture water to obtain a natural hirudin crude product; the method comprises the following steps: filtering by using a bag filter to remove fine particles, effectively separating hirudin from components larger than hirudin and components smaller than hirudin by using ultrafiltration membranes with different molecular weights, and hydrolyzing protein larger than hirudin in molecular weight by using neutral protease or alkaline protease; the leeches are extracted and separated to obtain various products such as hirudin, small-molecule oligopeptide and small-molecule extracts. According to the method, the culture pond is directly used for induction, emetic and separation, hirudin is effectively separated from macromolecules and micromolecules, combination can be carried out according to market requirements, and multi-variety and multi-effect leech series products are obtained.

The utility model discloses a hirudin extraction device convenient to clean, and relates to the technical field of hirudin extraction. Comprising a collecting box, a separating frame and a box cover, a first clip edge is arranged at the bottom end of the separating frame, the separating frame is clamped on the collecting box through the first clip edge, a stainless steel screen is installed in the separating frame, and a second clip edge is arranged at the bottom end of the box cover. According to the hirudin extraction device convenient to clean, living leeches are placed on a stainless steel screen, an electric heating plate is used for heating, the environment temperature in the box is kept within a suitable range, under irradiation of an ultraviolet induction lamp, an inducer is sprayed to the body surfaces of the leeches through a spraying pipe, and the leeches are stimulated to secrete secretory fluid in a warm white light flickering mode; due to the fact that the collecting box, the separating frame and the box cover are designed in a split mode, the living leeches can be conveniently cleaned while the living leeches are conveniently separated from the secretory fluid, follow-up cultivation and reutilization are facilitated, and operation is easy.

The present application relates to an exosome with high permeability and application thereof, the surface of the exosome is connected with dioleoyl phosphatidylserine, 1,2-dioleoyl-sn-glycerol-3-phosphoethanolamine and hirudin. The exosome constructed by the present application can improve the permeability of the skin, can carry active components directly to the skin epidermis and dermis, and accumulate in the epidermis and dermis, continuously provide nutrition for the skin, and fully exert the skin care and beauty effect.

The invention provides a formula of an aquatic animal poecilobdella manillensis healthy growth factor, and relates to the field of leech breeding. The formula of the healthy growth factor of the aquatic animal poecilobdella manillensis comprises the following raw materials: fresh livestock blood, tea leaf peptide, glacial acetic acid and sterile distilled water, and the raw materials comprise 90 kg of fresh livestock blood, 20 g of tea leaf peptide, 1 g of glacial acetic acid and 9.979 kg of sterile distilled water according to 100 kg of basic materials. The whole process is free of chemical poison residues, conforms to the green industrialization trend of medical poecilobdella manillensis and hirudin products, has multiple effects of environment optimization, activity protection, immune regulation, synergistic interaction and the like from cultivation to product full-chain energization, can improve the survival rate of poecilobdella manillensis and the yield of hirudin, can also enhance the safety and stability of the products, and can be used for preparing the medical poecilobdella manillensis. The economic and medical values of the poecilobdella manillensis industry are obviously improved.

This invention discloses a method for preparing hirudin-tanshinone B self-assembled nanoparticles for reversing foam cell regeneration, belonging to the field of nanomedicine formulation technology. The method includes: dispersing hirudin (HV) and tanshinone B (SaB) in pure water at a mass ratio; adding zinc chloride solution dropwise under constant temperature stirring at a mass ratio of zinc chloride to the total mass of hirudin and tanshinone B; using zinc ions as a bridging medium, driving hirudin and tanshinone B to self-assemble into nanoparticles through non-covalent interactions; and obtaining the nanoparticles after solid-liquid separation. This invention achieves efficient co-loading of peptides and polyphenolic active molecules through a metal ion coordination-driven self-assembly strategy. The resulting nanoparticles have uniform particle size and good stability, significantly reducing lipid deposition in foam cells and enhancing their cellular burial function, thus reversing the function of foam cells to normal macrophages.

The invention discloses an external preparation for treating cold-dampness stasis type old lumbocrural pain and a preparation method and application thereof, and belongs to the technical field of external preparations. The preparation comprises turpentine, a cortex pseudolaricis extract, modified hirudin dry powder, radix clematidis sapogenin dry powder and the like, and 60-degree pure grain white spirit is used as a matrix and matched with components such as a borneol composite inclusion compound and modified nanoscale magnet powder. Wherein the modified hirudin is obtained through synergistic embedding of liposome and licoflavone, the borneol composite inclusion compound is obtained through synergistic embedding of beta-cyclodextrin and hydroxypropyl methyl cellulose, the modified nanoscale magnet powder is obtained through grafting modification of silane-polyethylene glycol, and the modified nanoscale anhydrous mirabilite-polyethylene glycol compound is obtained through carboxymethylation modification. The preparation can specifically act on the pathological link of cold-dampness stasis type old lumbocrural pain, is beneficial to improving the transdermal absorption efficiency of the medicine and the stability of the preparation and reducing skin irritation, has a treatment effect on related diseases, and has a relatively good clinical application prospect.