93 results about "THROMBIN HUMAN" patented technology

The invention discloses a method for screening a non-natural nucleic acid aptamer of targeted thrombin based on streptavidin magnetic beads and application, and belongs to the technical field of biology. According to the method, an initial screening library is efficiently constructed through a primer extension technology, and a chemical modification SELEX technical process is simplified; a streptavidin magnetic bead separation technology is introduced, so that the screening specificity and efficiency are improved, the screening period is remarkably shortened, and the cost is reduced; by selecting non-natural nucleic acid modification sites and adopting a strategy of combining reverse screening and forward screening, high specificity and affinity of the aptamer are ensured, and limitation of an existing screening method is overcome; the screened non-natural nucleic acid aptamer shows high affinity and excellent stability, the aptamer with a specific structure and function is successfully obtained by optimizing the proportion of nucleoside triphosphate monomers and extension reaction conditions, and a wide application prospect is provided for the fields of drug development, biosensor preparation and the like. A new path is opened up for research and application of the nucleic acid aptamer.

The invention relates to the technical field of probiotic beverages, in particular to a hirudin peptide probiotic beverage with a vascular scavenger function, which is prepared from the following components in percentage by mass: 1 to 10 percent of hirudin peptide, 0.1 to 5 percent of probiotics, 2 to 5 percent of prebiotics, 1 to 5 percent of fruit juice, 0.1 to 2 percent of vitamin, 0.1 to 1 percent of mineral substance, 0.5 to 2 percent of dietary fiber, 0.1 to 1 percent of natural extract, 0.1 to 1 percent of sweetening agent, 0.01 to 0.1 percent of preservative and 70 to 90 percent of water, the hirudin peptide in the beverage serves as a core component of the beverage, has remarkable anticoagulation and thrombolysis effects and can activate a fibrin dissolution system in blood and inhibit the activity of thrombin, so that thrombosis is effectively prevented and treated, the occurrence risk of cardiovascular and cerebrovascular events is reduced, and the hirudin peptide can also expand blood vessels, increase blood flow and promote blood circulation. Microcirculation is improved, the symptoms of palpitation, chest distress, paralysis of hands and feet, dizziness and headache can be relieved, and blood circulation and thrombosis resistance are improved.

The invention belongs to the technical field of biomedical materials, and discloses microgel assembly powder capable of rapidly stopping bleeding as well as a preparation method and application of the microgel assembly powder. The microgel assembly powder is composed of microgel powder and assembly agent powder, and the mass ratio of the microgel powder to the assembly agent powder is (1: 10)-(10: 1); the microgel powder comprises carboxymethylated and sodium trimetaphosphate cross-linked dual-modified polymer powder, and the assembling agent powder is polyphenol modified polymer powder. The microgel assembly powder can quickly absorb moisture in blood to concentrate the blood after being in contact with the blood, meanwhile, rich ortho-phenolic hydroxyl groups on the assembly agent can immediately form a hydrogen bond network among microgel, red blood cells and platelets, and the microgel assembly powder is assembled into hydrogel with excellent mechanical strength and adhesion performance. In addition, the powder can load bioactive molecule thrombin, activate multiple blood coagulation cascade reaction and promote formation of a fibrous protein network and blood clots, and through the synergistic effect of multiple hemostasis mechanisms, the powder shows rapid and effective hemostasis performance.

The invention relates to the technical field of biology, and discloses a method for detecting thrombin production promotion capability of extracellular vesicles in a blood sample, which comprises the following steps: centrifuging the blood sample collected within 2 hours at 2500g at room temperature for 15 minutes, sucking upper platelet-deficient plasma and placing in a centrifuge tube, centrifuging at 2500g at room temperature again for 15 minutes, and sucking upper platelet-free plasma; centrifuging the obtained platelet-free plasma at 4 DEG C at 20000g for 1 hour; and sucking the upper-layer extracellular vesicle-free plasma for later use, taking the obtained extracellular vesicle precipitate as a to-be-detected sample, and detecting the thrombin generation promotion capability of the extracellular vesicles by using a thrombin generation analyzer. The method provided by the invention can be used for accurately detecting the capability of the extracellular vesicles in the blood sample for promoting thrombin generation, has the advantages of convenience in operation, easiness in obtaining the sample, noninvasiveness, no damage, high detection speed, low cost and the like, has important significance on thrombus risk assessment and prediction, and has better clinical application value.

The present invention discloses a spatially separated orthogonal translation system and its application. In order to enhance the specific translational expression of target genes in microbial hosts and reduce the mis-translation of background genes, the present invention provides a spatially separated orthogonal translation system. Specifically, first, the codons of the target gene are replaced with the TAG stop codon. Secondly, RTBA and thrombin F2 are selected as the mRNA-protein interaction system. Then, TyrRS / tRNA is heterologously expressed. This system can incorporate O-methyl-L-tyrosine into the amber stop codon UAG, and further fuse and express TyrRS and F2 with disordered proteins respectively. Finally, the translational selectivity of the target gene is increased to 4.01-fold. Finally, the N-acetylglucosamine epimerase AGE is modified to obtain a mutant AGE OMeY , whose specificity constant k cat / K m is increased by 86.0% compared with the wild type, and further, the specific translational expression of AGE OMeY is achieved in Bacillus subtilis cells by using the optimal orthogonal translation system, and the yield of the target product is as high as 2.1 g / L.

The invention relates to the technical field of medical instruments, in particular to a plugging device for femoral artery pseudoaneurysm and an operation method of the plugging device. The plugging device comprises a sheathing canal assembly, an air bag assembly and a thrombin conveying assembly, and the air bag assembly and the thrombin conveying assembly are arranged in the sheathing canal assembly; the air bag assembly comprises an air bag body and an inflation tube. The inflation tube penetrates through the air bag body and is communicated with the air bag body; the air bag assembly can move in the axial direction of the sheathing canal assembly and is inflated and expanded after the air bag body extends out of the head end of the sheathing canal assembly. The thrombin conveying assembly can eject thrombin through the head end of the sheath tube assembly. Compared with non-invasive compression, through the design of the high-compliance air bag, flexible plugging of the tumor neck and self-adaption to the shape and the size of the tumor neck, the pain feeling of a patient is effectively relieved, the sealing effect is enhanced, and the operation requirements for the thrombin injection speed and the dose precision are reduced. After the air bag body seals the tumor neck, a small amount of thrombin can be injected into the tumor body, and therefore the risk that the thrombin escapes to the femoral artery is avoided.

The present invention discloses an aptamer biosensor based on a covalent organic framework (COF), as well as its preparation method and application. The biosensor comprises a porous triazine-based COF and a DNA aptamer adsorbed on the surface of the porous triazine-based COF. The COF-based biomolecule detection platform provided by the present invention has potential fluorescence switch detection capabilities and can specifically bind to the aptamer. It can be used to detect ATP and thrombin with high detection selectivity and sensitivity.

A tissue regeneration multi-component biomaterial composition, including: a) a chitosan component having a number average molar mass (Mn) of about 32-47 kilodalton (kDa) and a degree of deacetylation (DDA) of about 80- 90%; b) a thrombin component present in an amount effective to accelerate the solidification of the chitosan component with a blood-derived product mixture; and c) a trehalose component present in a lyoprotectant effective amount.

The invention belongs to the technical field of polypeptide preparation and biological medicine, and particularly relates to a preparation method of Jingzhaotoxin polypeptide IX (JZTX-IX) and application of Jingzhaotoxin polypeptide IX in treatment of diabetes mellitus. A chilobrachys jingzhao toxin polypeptide IX coding gene, a 6 * His tag nucleotide sequence, a thrombin restriction enzyme cutting site nucleotide sequence and an MBP hydrotropy tag nucleotide sequence are connected to a prokaryotic expression vector for prokaryotic expression, and the expressed polypeptide is purified to obtain the chilobrachys jingzhao toxin polypeptide IX protein. The target polypeptide with high purity, complete disulfide bond folding and uniform conformation is obtained, so that the high purity and biological activity of the target polypeptide are ensured. On the basis, the treatment activity of the JZTX-IX on diabetes mellitus especially type II diabetes mellitus is found and verified, and the JZTX-IX can remarkably reduce fasting blood glucose of mice with the type II diabetes mellitus, improve glucose tolerance and insulin sensitivity, relieve polydipsia and polyuria symptoms and promote insulin secretion and islet cell proliferation.

The present disclosure is directed to human protease activated receptor 4 (PAR4) binding proteins (e.g. antibodies). In particular, anti-PAR4 binding proteins which are antagonists of human PAR4, as well as methods and uses thereof.

The invention relates to the technical field of thrombin detection, in particular to a thrombin fluorescent substrate applied to whole blood as well as a preparation method and application of the thrombin fluorescent substrate. The thrombin fluorescent substrate comprises a specific recognition peptide chain D-Phe (4-MeSO2)-Pro-Gly-Arg and a fluorescent group. The thrombin fluorescent substrate disclosed by the invention has high affinity with thrombin, high catalytic efficiency and strong anti-interference capability, and can be widely applied to a thrombin detection technology.

The invention discloses a recombinant leech polypeptide with high anticoagulant activity as well as a preparation method and application thereof. A PelB-T2Y-N47K-N52Y mutant is constructed by carrying out structural optimization on a hirudin HV2 sequence, a polypeptide product with relatively high anticoagulant activity is obtained after induced expression and nickel affinity chromatography purification are carried out in a prokaryotic expression system, and the anticoagulant activity of the polypeptide product reaches 9135.8 ATU / mg and is remarkably superior to that of an initial sequence HV2. Meanwhile, the polypeptide shows a lower IC50 value in a hydroxyl radical scavenging experiment, and has the activity of inhibiting generation of AGEs in a non-enzymatic saccharification model. Molecular docking and binding free energy analysis show that the combination of the mutant and thrombin (PDB ID: 4HTC) is more stable. The leech polypeptide disclosed by the invention has the potential of being developed into a multifunctional bioactive peptide preparation, and is suitable for the related fields of anticoagulation, antioxidation and anti-saccharification.

The invention belongs to the technical field of polypeptide preparation and biological medicine, and particularly relates to a preparation method of Jingzhaotoxin polypeptide XI (JZTX-XI) and application of Jingzhaotoxin polypeptide XI (JZTX-XI) in treatment of diabetes mellitus. A Jingzhaotoxin polypeptide XI coding gene, a nucleotide sequence of a 6 * His tag, a nucleotide sequence of a thrombin restriction enzyme cutting site and a nucleotide sequence of an MBP hydrotropy tag are connected to a prokaryotic expression vector for prokaryotic expression, and a target polypeptide which is high in purity, complete in disulfide bond folding and uniform in conformation is obtained through purification. The purity and the biological activity are relatively high. Meanwhile, the treatment activity of the JZTX-XI in the aspect of diabetes is verified, and it is found that the JZTX-XI can remarkably reduce fasting blood glucose of mice with type II diabetes, remarkably up-regulate a glucose transport regulation gene Glut2, improve glucose tolerance and insulin sensitivity, relieve polydipsia and polyuria symptoms and promote insulin secretion and islet cell proliferation.

The present disclosure provides synthetic compositions useful as inhibitors of mannan-binding lectin-associated serine protease-2 (MASP-2), including compositions that selectively inhibit MASP-2 over thrombin, as well as methods for their production and use.

The invention belongs to the technical field of hemostatic nursing devices for digestive tracts, and provides a bleeding nursing device for digestion. The clamping device comprises a clamping main body, wherein the clamping main body is provided with a clamping arm and an abutting disc support; the two clamping arms are oppositely hinged to each other; the number of the abutting disc supports is two, and the abutting disc supports are installed at one ends of the clamping arms correspondingly and hinged to the clamping arms. The hemostasis assembly is arranged on the clamping main body and is located on the abutting disc support; the hemostasis assembly comprises a plasma electrocoagulation needle, clamping teeth, seta and soft hairs; the blood coagulation assembly is arranged on the clamping main body; the blood coagulation assembly comprises a thrombin release microneedle; wherein the thrombin release microneedle is filled with thrombin, and the thrombin is used for rapid blood coagulation. The device can nurse bleeding spots which are not easy to fix, and the application range of the device is effectively widened, so that the device is suitable for industrial popularization.

The present invention discloses a method for promoting heart valve endothelialization and an artificial heart valve. The method is based on modifying circular DNA on the surface of a decellularized heart valve (DHV). A DNA hydrogel targeting thrombin is obtained on the surface of the DHV by rolling circle amplification. The DNA hydrogel not only exerts an anticoagulant function, but also acts as a heme capturer, recruiting heme from the blood. The heme is further converted into biliverdin (BV) by the heme oxygenase (HO1) loaded in the DNA, ensuring the continuous production of biliverdin, promoting anti-inflammatory immune regulation and reactive oxygen scavenging, thereby creating a regenerative immune microenvironment and promoting valve endothelialization.

The invention provides a compound of formula (I): (I) or a salt thereof, wherein R1-R2, L1-L2, A. B, C, x, and y have any of the values described in the specification, as well as compositions comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof. The compounds are useful as thrombin inhibitors.

The application discloses a thrombin solution virus inactivation device, which comprises a pump body one and a rack, the input end of the pump body one is communicated with a liquid storage device for storing thrombin solution through a liquid inlet pipe, the output end of the pump body one is provided with a liquid outlet pipe one, a protective shell is arranged at the middle position of the rack, a light transmission cylinder for placing the thrombin solution is arranged in the protective shell, a plurality of pairs of semicircular protrusions for stirring the liquid are arranged on the inner wall of the light transmission cylinder alternately, a secondary transmission gear is fixedly arranged at one end of the light transmission cylinder, the liquid outlet pipe one penetrates through the secondary transmission gear and extends into the light transmission cylinder, a plurality of ultraviolet lamps are arranged in the annular distribution between the light transmission cylinder and the protective shell, a driving motor is arranged at the top of the rack, a primary transmission gear corresponding to the secondary transmission gear is fixedly connected to the output end of the driving motor, and the primary transmission gear and the secondary transmission gear are transmissionally connected through a transmission belt, the semicircular protrusions arranged on the inner wall of the light transmission cylinder alternately can generate a turbulence effect on the thrombin solution in the light transmission cylinder when the light transmission cylinder rotates, the solution is promoted to be stirred more uniformly, the ultraviolet light can be refracted, the optical path length is increased, the ultraviolet light is more fully contacted with the solution, and thus the virus inactivation efficiency is improved.

The invention provides a method for efficiently expressing and purifying Taq DNA polymerase by using HEK293T, and belongs to the technical field of molecular biology. According to an amino acid sequence of Taq DNA polymerase, thrombin recognition and cleavage sites, six connected His amino acid tags and IFNalpha-2 signal peptides are sequentially designed at the N end of the Taq DNA polymerase, reverse translation is performed to obtain a nucleotide sequence, codons are optimized according to the environment of HEK293T cells, and then an eukaryotic expression vector is constructed. And transfecting the eukaryotic expression vector to an HEK293T cell to obtain the target protein. The purity of the obtained Taq DNA polymerase can reach 100%, the protein yield is high, about 2 mg of Taq DNA polymerase can be obtained by 20 mL of a culture medium, and the method is economical and efficient.

This invention provides 19 F-PDA-DNA-DTPA / Gd nanosensing probe, its preparation method, and its application in fluorine magnetic resonance imaging: This nanosensing probe consists of an MR signal source, a quencher, and a response linker. The MR signal source is a high-density fluorine perfluorocarbon nanoparticle, the quencher is Gd-DTPA, and the response linker is a DNA aptamer composed of a single-stranded DNA S1 and its complementary strand S2, with nucleotide sequences shown in SEQ ID NO.1 and SEQ ID NO.2. As a responsive nanoparticle platform, it is only visible upon activation by a specific biological trigger. Its biological application is demonstrated by targeting thrombin with the aptamer and exhibiting a strong and selective response to thrombin in live mice.

Disclosed herein are fusion proteins comprising a truncated thrombostasin protein having at least 85% sequence homology to a thrombostasin protein, wherein the thrombostasin protein has a carboxy terminal deletion; and a fusion partner protein that is a non-thrombostasin protein. Further disclosed are vaccine compositions thrombostasin proteins having a comprising a carboxy terminal deletion, and methods for inhibiting a response to a thrombostasin protein in a host in need thereof, comprising the disclosed fusion proteins or vaccine compositions. Further disclosed are methods for the preparation of a fusion protein composition.

The invention discloses a solid-state self-enhanced ECL aptamer thrombin sensor as well as a construction method and application of the solid-state self-enhanced ECL aptamer thrombin sensor. The method comprises the following steps: (1) synthesizing conjugated polymer points PDC; (2) incubating a PDC / dicarboxyl bipyridine ruthenium solid self-enhanced ECL working electrode; and (3) construction of the conjugate heme-G-quadruplex / AuNPs (AuNPs). The branched chain polyethyleneimine and tri-n-propylamine are similar in multi-tertiary amine structure, so that the branched chain polyethyleneimine and a polymer point PDC formed by hydrothermal synthesis of NH2BDC also have the capability of serving as a bipyridyl ruthenium co-reaction reagent, the detection range of the ultra-sensitive TB aptamer sensor of a sandwich structure constructed based on the PDC is 1015-108 M, the fluctuation range of TB in a blood sample is included, and the sensitivity of the TB aptamer sensor is improved. The detection limit is 3.6 * 10 <-16 > M, and the probe has an application prospect in thrombin medical detection.

The invention discloses a method for detecting polyvinyl chloride micro-plastic through photoelectrochemistry. Specifically, the invention develops a polyvinyl chloride micro-plastic photoelectrochemical aptamer sensor, and establishes a detection method based on the sensor, the method comprises the following steps: forming a double-chain compound by a thrombin aptamer P1 and a polyvinyl chloride aptamer P2, releasing P1 after the P2 recognizes PVC, splitting P1 under the action of exonuclease I, and detecting the PVC by virtue of the split P1. At the moment, free thrombin in the system degrades the p-aminophenol modified polypeptide to generate AP, a Bi < 3 + > / Bi < 2 + > MMCT process is triggered, enhanced photocurrent is output, AP regeneration is promoted through DI-mediated EDRR, the photocurrent is further amplified, and the sensitivity of a photoelectrochemical (PEC) sensor for detecting PVC is remarkably improved. The detection method is easy to operate, high in response speed, good in selectivity and high in sensitivity, and has good accuracy in detection of actual samples (such as cosmetics as lipsticks and liquid foundation).

Therapeutic antibodies specific for osteopontin and methods of using them for treating osteopontin-associated disorders are provided. In particular, antibodies, or antigen-binding fragments thereof, that inhibit thrombin-cleavage of osteopontin or block the activity of thrombin cleavage fragments of osteopontin are provided. Additionally, antibody conjugates and pharmaceutical compositions or formulations comprising the antibodies or antibody conjugates as well as kits including the antibodies, conjugates, or formulations are also provided.

The invention provides a method for improving the appearance of a freeze-dried antithrombin III product. The method comprises the following steps: carrying out pre-freezing treatment and sublimation drying treatment on a sample containing antithrombin III; the pre-freezing treatment comprises at least two cooling steps, and the temperature of each cooling step is-60 to-20 DEG C independently; the sublimation drying treatment comprises at least three heating steps, and the temperature of each heating step is-45 DEG C to 0 DEG C independently. The antithrombin III freeze-dried product obtained by the method disclosed by the invention meets the requirements of pharmacopoeia quality standards, and has the advantages of good molding, full appearance, no shrinkage, no collapse, no compact film layer at the top, high mechanical strength, good product stability and high market value, and is beneficial to storage and transportation.

Therapeutic antibodies specific for osteopontin and methods of using them for treating osteopontin-associated disorders are provided. In particular, antibodies, or antigen-binding fragments thereof, that inhibit thrombin-cleavage of osteopontin or block the activity of thrombin cleavage fragments of osteopontin are provided. Additionally, antibody conjugates and pharmaceutical compositions or formulations comprising the antibodies or antibody conjugates as well as kits including the antibodies, conjugates, or formulations are also provided.

The application belongs to the technical field of biotechnology, and particularly relates to a purification method of type A recombinant botulinum toxin. The purification method comprises the following steps: collecting a fermentation liquor, crushing a bacterial body to obtain a crushing liquor; adding a flocculating agent to the crushing liquor to clarify, centrifuging to remove impurities to obtain a clarified liquor; adding the clarified liquor to an ion chromatography column to perform ion chromatography to obtain an ion chromatography liquor; adding thrombin to the ion chromatography liquor to perform enzymatic cutting to obtain an enzymatic cutting liquor; adding the enzymatic cutting liquor to a hydrophobic chromatography column to perform hydrophobic chromatography to obtain a hydrophobic chromatography liquor; performing ultrafiltration concentration on the hydrophobic chromatography liquor to obtain an ultrafiltration concentrated liquor; adding the ultrafiltration concentrated liquor to a molecular sieve chromatography column to perform molecular sieve chromatography to obtain the purified type A recombinant botulinum toxin. Compared with the prior art, the purification method can effectively remove impurity proteins, does not affect the integrity of light chains and heavy chains, has an amino acid sequence coverage of 100%, and does not affect the amino acid sequence coverage of 100%.

The application relates to the technical field of biological medicine, and particularly discloses a kit for determining antithrombin III activity, which comprises a first reagent, a second reagent, a third reagent and a diluent; the first reagent at least comprises a first buffer, thrombin, a serine protease inhibitor and a first stabilizer; the second reagent at least comprises a second buffer, a chromogenic substrate and a second stabilizer; the third reagent at least comprises a third buffer, heparin, isopropyl alcohol and a third stabilizer; and the diluent at least comprises physiological saline and a fourth stabilizer. In the kit, the cost is low, the anti-interference capability is strong, the sensitivity is high, the stability is good, and the kit has a wide application prospect.

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.

This invention provides a gene encoding a coral-derived anticoagulant polypeptide and its applications, belonging to the field of bioactive peptide technology. The invention provides an anticoagulant polypeptide GcKuz1, which includes the mature peptide segment shown in SEQ ID No. 1. The polypeptide GcKuz1 of this invention exerts antithrombotic and anticoagulant effects by binding to KLKB1 in the thrombin system. This polypeptide does not cause hemolysis or cytotoxicity, and has no bleeding risk, exhibiting good biocompatibility. The anticoagulant polypeptide of this invention has therapeutic effects on thrombotic diseases, such as arterial thrombotic diseases, venous thrombotic diseases, and capillary thrombotic diseases.