45 results about "Basophilic Granulocyte" patented technology

On one hand, the invention provides a compound with a structure of general formula I or a conjugate thereof, wherein each group is as defined in the description. On the other hand, the invention provides a composition comprising (i) the compound of formula I or the conjugate thereof, and (ii) at least one surfactant selected from cationic surfactants and nonionic surfactants. The invention also provides a preparation method of the composition and a kit containing the composition. The invention still provides a method for the simultaneous identification of nucleated erythrocyte, basophilic granulocyte and lymphocyte and the classification by using the composition disclosed in the invention.

The invention relates to the technical field of medical examination and discloses a hemolytic agent and application thereof as well as a classifying and counting method for white blood cells. The hemolytic agent contains one or more than two kinds of cationic surfactants, one or more than two kinds of small molecule alcohols, one or more than two kinds of buffer solutions, one or more than two kinds of soluble inorganic salts and the balance of water. According to the invention, components of the hemolytic agent are simplified; the novel hemolytic agent suitable for classifying and counting basophilic granulocyte and other white blood cells is prepared by adopting the raw materials which are proper and easy to obtain; compared with the current hemolytic agent product with more complex components, the hemolytic agent disclosed by the invention is more convenient and quicker for use; the hemolysis time only needs 5-20s, the classifying and the counting for the basophilic granulocyte and the other white blood cells can be realized by combining the hemolytic agent with an impedance analysis measuring instrument without special requirement on the temperature, and the accuracy is higher.

The invention relates to compounds of general formula (I); 1a, 1 b, 1 c and 1 d. The invention also relates to a method for the production thereof, pharmaceutical preparations containing said compounds and / or physiologically compatible salts thereof which can be produced therefrom and / or solvates thereof, and to the pharmaceutical use of said compounds, salts or solvates thereof as inhibitors of phosphodiesterase 4. The compounds comprise active ingredients for the treatment of diseases which can have a positive influence by inhibiting the activity of phosphodiesterase 4 and / or TNFα-release, for example, in lymphocytes, eosinophile and basophile granulocytes, macrophages and mastocytes.

The invention discloses a leukocyte classification and a classification method thereof, and the kit is composed of a water soluble quantum dots-cyanine dye probe, and an erythrocyte and platelet hemolytic agent, wherein the quantum dots-cyanine dye probe is composed of a stabilizing agent, which takes a polypeptides material as the quantum dots, and Cy5 cyanine dye covalent joint molecules whose emission spectrum is in the near infrared area, and has the characteristics of high quantum yield and sensitivity; the erythrocyte and platelet hemolytic agent can rapidly solves erythrocytes in the blood, and is composed of a cationic surfactant, a non-ionic surfactant, a pH buffer agent or combination of the cationic surfactant, the non-ionic surfactant, and the pH buffer agent. The leukocyte classification reagent provided by the invention can carry out precise subgroup identification on lymphocyte, monocyte, neutrophil, basophilic granulocyte, and acidophilic granulocyte in leukocytes of blood samples, and further identify immature cells or abnormal cells.

The invention relates to a paper based sensor capable of detecting allergic reactions as well as preparation and application of the paper based sensor and belongs to the field of paper based sensors.The allergic reaction refers to abnormal and excessive immune response caused by the reasons that a specific immunoglobulin (IgE) antibody is produced when the body is stimulated by a certain antigenand the IgE can be bound to an IgE specific receptor on the surface of mast cells and basophilic granulocyte. Therefore, the IgE in serum is an important marker for detecting the allergic reaction. Ni-NTA can be specifically bound with a His label, thereby realizing affinity purification of proteins. The paper based material has the advantages of being cheap, portable, high in selectivity and biocompatibility and the like, so the paper based sensor containing the Ni-NTA is prepared and can be firmly bound to an allergen containing the His label, and qualitative detection of the immune globulinis realized. The paper based sensor has important significance for rapidly and sensitively detecting the allergic reactions.

The invention provides a basophilic granulocyte activation and degranulation identification method. The basophilic granulocyte activation and degranulation identification method comprises the following steps that test samples are numbered in sequence, and flow type sample feeding pipes required by testing are marked; a required anti-human CD123, anti-human CCR3, anti-human HLA-DR, anti-human CD203c and / or anti-human CD63 fluorescence labeling flow type antibody combination is added into each flow type sample feeding pipe; mixed whole blood is added into the marked flow type pipes, and shading incubation is conducted at the indoor temperature; red blood cell lysate is added into each sample pipe, and shading incubation is conducted at the indoor temperature again; supernatant is removed in a centrifugal mode after incubation; detection is conducted by means of a flow cytometry, and the number of basophilic granulocytes and the average fluorescence intensity are analyzed. According to the basophilic granulocyte activation and degranulation identification method, 80%-100% of the basophilic granulocytes in peripheral blood can be distinguished from other types of cells under the condition that the basophilic granulocytes do not need to be separated or purified, the basophilic granulocyte activation and / or degranulation state can be identified, whole blood of no more than 100 microlitres is needed by each sample to be tested, and the repeatability is good.

The invention discloses a method for detecting an anaphylactoid reaction of a Shengmai liquid preparation. According to the method, the quality of a finished Shengmai liquid preparation product is detected by inducing the activation amount of basophilic granulocyte through the Shengmai liquid preparation, and the method is a quick detection method for industrial scale production safety evaluation of the Shengmai liquid preparation, so that the quality of the finished Shengmai liquid preparation product is ensured, and the occurrence rate of the anaphylactoid reaction in the clinical application process is reduced.

The invention discloses a hemolytic agent for leukocyte and basophilic granulocyte counting and hemoglobinometry, and belongs to the technical field of the blood analysis. The problem that the leukocyte and basophilic granulocyte counting and hemoglobin in the human whole blood cannot be determined at the same time in the prior art is solved. The hemolytic agent is a buffer solution containing cationic surfactant, a nonionic surfactant and hemoglobin stabilizer, wherein the concentration of the cationic surfactant is 0.1g / L to 50g / L, the concentration of the nonionic surfactant is 0.5g / L to 100g / L, the concentration of the hemoglobin stabilizer is 0.1g / L to 50g / L, and a pH value of the buffer solution is 1.8-4.8. The component of hemolytic agent is simplified, the hemolysis time only needs10-18s without using a heating device, namely accomplishing the classified counting on the leukocyte and basophilic granulocyte and the determination on the hemoglobin; the operation is simple and fast and high in accuracy.

The invention discloses a reagent for classifying leukocytes, which can be used for classifying leukocytes into five groups corresponding to lymphocytes, monocytes, eosinophilic granulocytes, neutrophilic granulocytes and basophilic granulocytes. The reagent comprises a buffering agent and a hemolytic agent, wherein the hemolytic agent comprises at least two types of betaine type amphoteric surfactants. The reagent can be applied in wide pH value and temperature ranges, does not damage the leukocytes and can be used for accurately classifying white blood cells.

The invention relates to the field of medical diagnostics, and in particular relates to an application of granulocyte group basophils as diagnostic markers for allergic diseases. The inventors classify basophils into mononuclear cell group basophils and the granulocyte group basophils, and conduct a systematic study on the granulocyte group basophils to find that the granulocyte group basophils are specifically associated with the allergic diseases and can be used for the diagnosis of the allergic diseases. The diagnostic method has good specificity and easy detection, and is easy to promote and clinically apply.

The invention discloses an anti-human FcepsilonRIalpha subunit monoclonal antibody and application thereof. A hybridoma cell strain producing the monoclonal antibody is collected in China Center for Type Culture Collection, and the collection number thereof is CCTCC NO:201211. The anti-human FcepsilonRIalpha subunit monoclonal antibody prepared by the invention can be combined with a human IgE high-affinity receptor FcepsilonRIalpha, and can be used for detecting FcepsilonRIalpha expression on the surface of a cell; and the monoclonal antibody is of great significance for prevention and treatment of mast cell and basophilic granulocyte related diseases such as allergic dermatitis, dermatomyositis, psoriasis and the like, and has great clinical application value. The hybridoma cell strain provided by the invention is favorable in cell strain growth vigor, uniform in cell body size, muddy or transparent and vigorous in division, and has a proliferative potential of infinite division.

The invention relates to a method used for detecting ostrea plicatula eosinophilic and basophilic granulocyte phagocytic ability, and the method comprises (1) ostrea plicatula blood cell suspension preparation; (2) fluorescent microsphere phagocytosis; (3) staining with neutral red; (4) lining dying with propidium iodide; and (5) microscopy. The method has the advantages of being simple in operation, rapid and accurate, strong in specificity, high in repeatability, and the like.

The present invention concerns a method for determining the degranulation activity of basophilic granulocytes in a sample as well as a reagent kit that is suitable for this. Furthermore the invention also concerns a method for diagnosing allergic hypersensitivity and for monitoring the response to a hyposensitization therapy.

The invention relates to a cell detection kit and application thereof. The cell detection kit includes a staining solution and a hemolytic agent. Nucleated red blood cells, basophilic granulocyte andother four kinds of white blood cells are distinguished through simultaneous usage of reagents in the cell detection kit, the nucleated red blood cells and the basophilic granulocyte can be accuratelydetermined, the basophilic granulocyte and the other four kinds of white blood cells can be distinguished through individual usage of the hemolytic agent, and the basophilic granulocyte can be accurately determined. Therefore, according to the cell detection kit, the basophilic granulocyte and / or the nucleated red blood cells and other white blood cells can be classified and counted through autonomous selection of modes.

The invention discloses a construction method of a transcriptome expression profile of activated basophilic granulocyte. The construction method comprises the following steps of: preparing, activatingand culturing basophilic granulocyte, extracting total RNA of cells, sequencing a whole transcriptome and identifying a whole transcriptome expression profile. The invention further discloses the transcriptome expression profile of the activated basophilic granulocyte; compared with a transcriptome expression profile of non-activated human basophilic granulocyte, 6 remarkable up-regulation effects exist in mRNA (messenger ribonucleic acid), 10 remarkable up-regulation effects exist in lncRNA (long non-coding ribonucleic acid), 6 remarkable up-regulation effects exist in circRNA (circular ribonucleic acid) and 8 remarkable up-regulation effects exist in microRNA. According to the construction method disclosed by the invention, the change of the transcriptome expression profile related to the activation of the human basophilic granulocyte is systematically researched and a plurality of RNA which are differentially expressed after being activated are identified; and the RNA provides a new target for inhibiting the activation of the human basophilic granulocyte, so that a new direction is provided for treating allergic diseases and autoimmune diseases.

The invention provides a method for detecting the sensitivity of suspected allergen in milk and dairy products. The method mainly comprises the following steps: preparing the milk or the dairy products to be detected into sample solution, mixing with human basophilic granulocyte, culturing, centrifuging and detecting the amine content of a supernatant group. The method of the invention realizes food sensitization detection at cell level, and is effective to replace a clinical skin-prick test (skin test).

The invention provides a hemolytic agent applied to differential counting of white blood cells. The hemolytic agent contains a cationic surfactant capable of effectively dissolving red blood cells; the hemolytic agent contains a nonionic surfactant; the nonionic surfactant can cooperate with the cationic surfactant to crack the red blood cells, can rapidly break red blood cell membranes and can completely dissolve the red blood cells and enable the volume of the red blood cell fragments to be reduced so as to cause no interference on the analysis of the white blood cells; trifluoromethanesulfonic acid is added, so that the ductility of the membranes can be reduced, and leakage and hemolysis of the red blood cells are caused; the stability and the dispersibility of the white blood cells inblood samples can be improved; meanwhile, the hemolysis effects of the surfactants can be improved; the surfactants supplement each other, so that the differential counting number of the white blood cells is more accurate; a phosphate buffer solution and disodium edetate dehydrate are added, so that basophilic granulocytes and other white blood cells can be accurately distinguished; sodium chloride is added, so that the integrity of the state of the white blood cells in an isosmotic solution is retained. The hemolytic agent is non-toxic and environmentally friendly, and is excellent in hemolytic effect, high in dispersibility of the blood samples after hemolysis, convenient in subsequent operations, free of damage to instruments and free of harmless treatment on waste liquid.

The invention discloses a test method for basophilic granulocyte activation and degranulation. The method comprises: diluting a to-be-tested allergen to prepare a standard to-be-tested allergen, setting a positive control and a negative control, wherein all to-be-detected samples are heparinized peripheral blood of a patient and need to be sealed and incubated, then adding a buffer solution, centrifuging, adding a fluorescent dyeing mixed solution, lightly mixing until precipitate is suspended, incubating in a dark place, adding an erythrocyte lysate, uniformly mixing in a vortex manner, incubating in a dark place, diluting, centrifuging, re-suspending the precipitate, adding a buffer solution after filtering, and detecting by adopting a flow cytometer to obtain an activation and degranulation ratio. According to the method, the basophilic granulocyte is not required to be separated and purified, the new specific basophilic granulocyte marker combination is selected, the use amount ofthe detected sample is small, the repeatability is good, the error is small, the activation and degranulation degrees of the basophilic granulocyte are effectively distinguished, and the detection andthe technical support are well provided for the clinical differential diagnosis and the scientific research project of related diseases.

The invention discloses an analytical reagent of basophils, which comprises at least one alkyl phenol polyoxyethylene ether, at least one alkyl alcohol polyoxyethylene ether and a buffering agent for adjusting a pH (pondus Hydrogenii) value. The invention also discloses a method of adopting the reagent to determine the basophils. The reagent of the invention can crack red blood cells and platelets, damage cell membranes of other white blood cells except the basophils, and keep the structure integrity of the basophils so as to sort and count the basophils through optical signals determining the sizes and the internal structures of the cells.

The invention relates to a ragweed pollen allergen extract, ragweed pollen allergen extraction liquid and a preparation method of the extract. The ragweed pollen allergen extraction liquid includes ragweed pollen allergens containing the amino acid sequences shown in SEQIDNO.4 to SEQIDNO.9. The ragweed pollen allergen extraction liquid is prepared through ragweed pollen collection, drying, degreasing, extraction, ultra-filtration concentration, freeze drying, re-dissolving and the like and mainly includes ragweed pollen allergens, glycerinum, sodium chloride and the like. The ragweed pollen allergen extraction liquid has the advantage of being high in specificity, allergenic protein components of ragweed are fully extracted, the total biological value is stable, the service life is long, and the sterile effect is good. The stoste of the ragweed pollen allergen extraction liquid can be effectively used for skin prick test diagnosis of inspiratory allergosis, in-vitro basophilic granulocyte activation test diagnosis, intracutaneous test diagnosis and specific immunotherapy after appropriate dilution, and can effectively diagnose inspiratory allergosis induced by ragweed pollen and conduct specific immunotherapy on the inspiratory allergosis.

The invention relates to a method for differentiating a shellfish granular blood cell types. The method comprises the following steps of: (1) dissolving neutral red in a phosphate buffer solution at a mass ratio of 100:(1-3), filtering and then storing at a temperature of 4-5 DEG C, and diluting before use; (2) preparing a shellfish blood cell suspension, taking 1-2 ml of a neutral red working solution and 1-2 ml of the blood cell suspension and mixing evenly in a cell culture plate hole, dyeing for 30-40 minutes, then dropwise adding a drop of evenly mixed solution on a glass slide, and then observing the type of the shellfish granular blood cells through microscopic examination. Compared with the traditional MGG (May-Grunwald -Giemsa) dyeing method, the method provided by the invention is free of cell fixation, less in operation to the cells, and capable of clearly differentiating the granular blood cell type; therefore, a very convenient and practicable method is provided for studying the phagocytic capability, athletic capability, reactive oxygen generation capability and wound repair capability of eosinophilic granulocyte and basophilic granulocyte from the aspect of living cells.

The invention discloses application of IL-1ra-Fcepsilon fusion gene and coded protein thereof to treatment on allergic rhinitis, belonging to the field of medicines. In the invention, the constructed IL-1ra-Fcepsilon fusion gene expression vector is prepared into suspension according to a certain concentration and the suspension is directly dropped into a nasal cavity to treat the allergic rhinitis; or the purified IL-1ra-Fcepsilon fusion gene is diluted to a certain concentration with a PBS (Phosphate Buffered Saline) buffer solution and then the diluted solution is dropped into the nasal cavity to treat the allergic rhinitis. In the invention, the protein coded by the IL-1ra-Fcepsilon fusion gene can antagonize the inflammatory effect of IL-1 and inhibit mastocyte and basophilic granulocyte degranulation, thereby achieving the aim of treating the allergic rhinitis.

The invention discloses an anti-human Fc epsilon RI alpha subunit monoclonal antibody and application thereof. A hybridoma cell strain capable of producing the monoclonal antibody is preserved in China Center for Type Culture Collection and has the preservation number of CCTCC NO. C201326. The anti-human Fc epsilon RI alpha monoclonal antibody prepared by the invention can be bound with high-affinity receptor Fc epsilon RI alpha of human IgE and can be applied in detecting the expression of cell surface Fc epsilon RI alpha. The monoclonal antibody has important significance and relatively large clinical value for prevention and treatment of mast cell and basophilic granulocyte-related diseases, such as allergic dermatitis, dermatomyositis and psoriasis. The hybridoma cell strain disclosed by the invention grows well, and is bright and transparent, the cell body is uniform, the division is strong and the hybridoma cell strain has indefinitely divided proliferative capability.

The invention provides a method for measuring free and cell binding total IgE in human whole blood and a diagnostic kit preparation method. The IgE is synthesized and secreted by plasma cells, most ofthe IgE and cells (such as basophilic granulocyte and mastocyte) are bound through receptors of the IgE, and only a little IgE is free in blood serum. An existing IgE detection method can only detectfree IgE in the blood serum and cannot detect the IgE bound on cell membranes. According to the method, blood pH (potential of hydrogen) is reduced, so that the IgE and the receptors of the IgE positioned on cells are dissociated, and the free and cell binding total IgE is detected by an ELISA (double-antibody one-step sandwich enzyme-linked immuno sorbent assay) method. The method overcomes theshortcoming that the existing IgE detection method can only detect the free IgE in the blood serum, and the free and cell binding total IgE can be detected.

The invention provides a Humulus japonicas pollen allergen extract, infusion liquid and a preparation method thereof. The Humulus japonicas pollen allergen extract contains an amino acid sequence of the Humulus japonicas pollen allergen protein shown as SEQ ID NO. 4-SEQ ID NO. 20. The infusion liquid is prepared by the processes of collecting of Humulus japonicas pollen, drying, degreasing, extracting, ultrafiltration concentrating, freeze drying, and redissolving. The Humulus japonicas pollen allergen infusion liquid has the characteristics of high specificity, the Humulus japonicas allergenic protein component is fully extracted, the total biological titer is stable, the effective period is long, and the sterilizing effect is good; an original solution can be effectively used for skin prick test diagnosis of allergic diseases, and can be used for in-vitro basophilic granulocyte activation test diagnosis, intradermal test diagnosis and specific immunotherapy after appropriate dilution(10-2-10-20), The infusion liquid can effectively diagnose allergic diseases induced by Humulus japonicas pollen and performs specific immunotherapy.

The invention discloses a hemolytic agent for a blood cell analyzing device. The hemolytic agent can decompose erythrocytes and blood platelets in blood. The invention also discloses a leukocyte classification kit. The leukocyte classification kit comprises the hemolytic agent, a dilute and an inhibitor. The agent provided by the invention is the assorted agent suitable for XFA9500 fully-automatic five-classification blood cell analyzer produced by Nanjing Perlong medical equipment limited company; the agent contains no dye, so that the agent is environment-friendly; by adopting the agent, lymphocyte, monocyte, neutrophile granulocyte, eosinophilic granulocyte and basophilic granulocyte can be directly detected; furthermore, the analysis method is fast and accurate.

The invention relates to the technical field of in-vitro diagnosis, and particularly discloses a reagent and a kit for classifying nucleated red blood cells, basophilic granulocytes and other white blood cells and application of the reagent and the kit. The reagent for classifying nucleated red blood cells, basophilic granulocytes and other white blood cells comprises a first reagent and a second reagent, the first reagent comprises a buffer agent, organic acid and a surfactant; the second reagent comprises a fluorescent dye and an organic solvent for dissolving the fluorescent dye. By means of the mode, nucleated red blood cells, basophilic granulocytes and other white blood cells can be distinguished, and then the nucleated red blood cells, the basophilic granulocytes and the other white blood cells are accurately counted.

The invention discloses application of IL-1ra-Fcepsilon fusion gene and coded protein thereof to treatment on allergic rhinitis, belonging to the field of medicines. In the invention, the constructed IL-1ra-Fcepsilon fusion gene expression vector is prepared into suspension according to a certain concentration and the suspension is directly dropped into a nasal cavity to treat the allergic rhinitis; or the purified IL-1ra-Fcepsilon fusion gene is diluted to a certain concentration with a PBS (Phosphate Buffered Saline) buffer solution and then the diluted solution is dropped into the nasal cavity to treat the allergic rhinitis. In the invention, the protein coded by the IL-1ra-Fcepsilon fusion gene can antagonize the inflammatory effect of IL-1 and inhibit mastocyte and basophilic granulocyte degranulation, thereby achieving the aim of treating the allergic rhinitis.

The invention discloses halogenated diarylurea compounds and application thereof in preparation of an antiallergic drug, and discloses for the first time that the halogenated diarylurea compounds caneffectively antagonize beta-aminohexylglycosidase release and histamine release of human basophilic granulocyte KU812 caused by C48 / 80, so as to inhibit anaphylactoid reaction. Therefore, when the halogenated diarylurea compounds are used as an anti-allergic preparation, the types of anti-allergic medicines are enriched, and more possible treatment schemes are provided for clinical anti-allergic treatment.