60 results about "Peripheral blood cell" patented technology

The invention discloses an application of CD4 + CD8 + double-positive T cells in improvement of poor immune reconstitution of HIV (human immunodeficiency virus) infected patients, and relates to the technical field of biological pharmacy, in particular to the application of preparation of the CD4 + CD8 + double-positive T cells in the poor immune reconstitution of the HIV infected patients. The number of peripheral blood CD4 + T cells of HIV infected patients is increased, the immune system of the patients is improved, and the probability of possible opportunity infection and other related diseases of the patients is reduced, so that the morbidity and mortality of the patients are reduced.

The invention discloses a Wright staining peripheral blood cell accurate identification system and method based on intelligent microscopic imaging and multi-modal fusion deep learning, belongs to the crossing field of medical examination and artificial intelligence, and is suitable for cell morphology analysis under a 10 * 40-time microscope. The core scheme comprises the following steps: standardized slide preparation and dyeing (the slide pushing angle is 25 + / -2 degrees, the pH is 6.8 + / -0.2, and color sensing dynamic regulation is performed); intelligent imaging (focusing formula and color normalization; the method comprises the following steps of (1) data processing, (2) data processing, (3) data processing, (4) data processing, (5) data processing, (6) data processing, (7) data processing, (8) data processing, (7) data processing, (8) data processing, (8) data processing, (8) data processing, (8) data processing, (8) data processing and (8) data processing.

The invention provides an ischemic stroke auxiliary diagnosis system based on a peripheral blood T cell single cell transcriptome and application thereof, and the auxiliary diagnosis system comprises a peripheral blood mononuclear cell acquisition module, a single cell RNA sequencing module, a data processing module, a data analysis module and a result discrimination module. With a coding gene of a protein molecule significantly related to the occurrence of ischemic stroke as a target gene, a data processing module obtains the expression level of the target gene in each T cell; the data analysis module obtains a risk score of each T cell of the subject by using a built-in single cell risk scoring model, and weights the risk scores of all the T cells through distributed perception identification to obtain an individual risk score of the subject; and the result judgment module is used for classifying the subjects into ischemic stroke negative and ischemic stroke positive according to the individualized risk scores of the subjects. The auxiliary diagnosis system disclosed by the invention is high in ischemic stroke discrimination capability, and high in sensitivity and specificity.

The invention discloses a biomarker for early warning or diagnosis of early-stage PD (Parkinson's Disease) and application thereof, belongs to the technical field of biological medicines, and proposes that the level of peripheral blood P-LCK cells in a blood sample is taken as the biomarker for the first time, and the biomarker has extremely high accuracy, specificity and sensitivity for early warning or diagnosis of early-stage PD.

The invention discloses application of 5 beta-cholanic acid (5 beta-CA) in enhancement of NK cell functions and preparation of antiviral drugs. The NK cell function of a chronic hepatitis B (CHB) patient is depleted, it is found that the intracellular 5beta-CA level is reduced by screening differential metabolites of peripheral blood NK cells of the CHB patient, and the NK cell function of the CHB patient can be enhanced by exogenous supplementation of 5beta-CA; the function of NK cells of HBV persistently infected mice can be enhanced, and the level of virology indexes such as serum HBV related antigens and the like can be reduced. Specifically, the 5beta-CA enhances the NK cell function by activating a TGR5-cAMP-mTOR signal channel so as to play an anti-HBV role. The research finds that the improvement of the 5beta-CA level is expected to become a new thought for enhancing the NK cell function of a CHB patient, and a new technical means is provided for the treatment of CHB.

The present application relates to the application of Bifidobacterium longum subsp.longum BL21 in the preparation of a preparation for improving hematopoietic dysfunction, wherein the Bifidobacterium longum subsp.longum BL21 is a Bifidobacterium longum subsp.longum BL21 strain with a preservation number of CGMCC No.10452.The present application develops a new microbial strategy for improving hematopoietic dysfunction, and uses the Bifidobacterium longum subsp.longum BL21 in the preparation of a preparation for improving hematopoietic dysfunction.It is found that the Bifidobacterium longum subsp.longum BL21 can increase the number of peripheral blood cells (red blood cells and white blood cells), the content of platelets or the content of hemoglobin, reverse the weight loss caused by hematopoietic dysfunction, and improve the serum inflammatory response and the spleen inflammatory response caused by hematopoietic dysfunction.The present application also provides a new idea for the wide application of the Bifidobacterium longum subsp.longum BL21.

The invention provides a method for regulating and controlling proliferation and movement of human peripheral blood T cells by Amuc1100 protein. The method relates to the technologies of separation, activation and culture of human peripheral blood T cells, CCK-8 cell proliferation detection, Transwell cell chemotactic detection, flow cytometry receptor expression body detection and the like. The application of the Amuc1100 protein to treat T cells proves that the protein can significantly down-regulate the expression of CD25 and CXCR4 of the T cells and inhibit T cell proliferation and CXCL12 induced T cell chemotactic movement respectively. The method and research means for in-vitro regulation of T cell proliferation and chemotactic movement can be applied to targeted regulation of T cells and directional migration of intestinal tracts of the T cells, and are used for maintaining the steady state of the intestinal tracts, relieving intestinal inflammations and screening drugs to treat diseases related to the intestinal inflammations.

The invention belongs to the technical field of tumor immunotherapy, and relates to an anti-Claudin18.2 single-domain antibody, a multifunctional fusion protein, a recombinant natural killer cell (CT-CAR-NK), and preparation and application thereof. Through alpaca immunization and phage display library construction and panning, the single-domain antibody VHH-C18.2-1 specifically combined with Claudin18.2 is obtained, and the amino acid sequence of the single-domain antibody VHH-C18.2-1 is SEQ ID NO: 1. The amino acid sequence of the designed fusion protein is SEQ ID NO: 3, the fusion protein sequentially comprises a VHH-C18.2-1, a flexible Linker, a TGF-beta RII extracellular domain, a CD8alpha hinge region, a CD8alpha transmembrane region, a 4-1BB intracellular domain and a CD3zeta intracellular domain from the N end to the C end, and the fusion protein has the functions of targeting, resisting TGF-beta inhibition and activating signals. The fusion protein gene transfects human peripheral blood CD56 + CD3-NK cells through lentivirus to obtain CT-CAR-NK, in-vitro verification shows that the CT-CAR-NK still keeps efficient killing in an immunosuppression environment, tumor growth can be remarkably inhibited in vivo, the lifetime can be prolonged, and a safe and efficient scheme is provided for Claudin18.2 positive solid tumor treatment.

The invention relates to a quality control substance for flow cytometry HLA-B27 analysis as well as a preparation method and application of the quality control substance. The quality control substance comprises a negative quality control substance and a positive quality control substance; the negative quality control substance comprises human lymphocyte negative quality control substance simulation particles and human peripheral blood cells or simulation particles; an HLA-B27 antigen is not expressed on the surface of the human lymphocyte negative quality control substance mimic particle; the positive quality control substance comprises human lymphocyte positive quality control substance simulation particles and human peripheral blood cells or simulation particles; an HLA-B27 antigen is expressed on the surface of the human lymphocyte positive quality control substance simulation particle; cD3 antigens and CD45 antigens are expressed on the surfaces of the human lymphocyte positive quality control substance simulation particles and the human lymphocyte negative quality control substance simulation particles. The technical problem to be solved is how to provide a quality control substance which is good in stability and can be applied to flow cytometry HLA-B27 analysis.

Provided in the present invention is a grading method for evaluating the severity of the clinical symptoms of hypersplenism. In the present invention, a model for evaluating the severity of the clinical symptoms of hypersplenism is constructed by means of performing logistic regression analysis on PLT, WBC and RBC to establish a nomogram, converting regression coefficients into a visual scoring system, separately performing score assignment on the basis of primary and secondary factors, and finally constructing a total-score-based hypersplenism grading prediction model. The model in the present invention is used to evaluate the severity of the clinical symptoms of hypersplenism in patients. By means of correlation analysis of peripheral blood cell testing results of patients with hypersplenism, hypersplenism grades of the patients are evaluated, such that personalized treatment plans can be provided to the patients in a timely manner, thereby avoiding delays in treatment.

The application belongs to the technical field of bioinformatics analysis, and discloses a kind of anti-new coronavirus neutralizing antibody VJ gene preference analysis method, including antibody database acquisition data, data screening, selection frequency statistics, selection preference analysis, selection frequency and preference visualization, pairing frequency statistics, pairing preference analysis, pairing frequency and preference visualization.The beneficial aspects of the present application are that the neutralizing antibody and VJ selection and pairing frequency are tested by scientific statistical methods, and are visualized to make the result interpretation more simple and intuitive.Therefore, according to the above method, a new method for early prediction of anti-new coronavirus neutralizing antibody production based on peripheral blood B cell VJ gene frequency and preference is established, which can predict whether the probability of producing neutralizing antibody by the subject is higher than the baseline level of healthy people.

This useful solution, in the field of biotechnology and regenerative medicine, relates to a process for enhancing the efficiency of induced pluripotent stem cell (iPSC) colony formation from human peripheral blood mononuclear cells (PBMCs). The solution aims to overcome the low reprogramming efficiency, unstable colony formation, and limited iPSC cloning rates commonly encountered in PBMC iPSC generation processes. The technical essence of the solution lies in the synchronized optimization of the reprogramming process, including pre-activating PBMCs with an appropriate cytokine combination at predetermined times before reprogramming, using a non-integrated Sendai virus system under optimized transformation conditions, switching culture media at defined intervals to promote cell state transitions, and applying a selection, enrichment, and stabilization process for iPSC cloning in the initial passages after colony formation.The process also incorporates cell line quality control steps through assessment of residual Sendai virus RNA removal and mycoplasma contamination testing during line maintenance. Experimental results show that the proposed process increases the number of iPSC colonies formed, improves the rate of establishing stable iPSC lines, and enhances cell homogeneity and pluripotency maintenance. The solution can be applied in basic research, pathogenesis modeling, drug screening, regenerative medicine, and studies using induced pluripotent stem cells derived from human peripheral blood cells.

The invention relates to application of a traditional Chinese medicine composition in preparation of a medicine for preventing or treating myelodysplastic syndrome and related complications thereof. The traditional Chinese medicine composition comprises sculellaria barbata, astragalus membranaceus, angelica sinensis, nightshade, radix curcumae, red ginseng, duchesnea indica, saussurea involucrata, cyclea verticillata and salvia miltiorrhiza. The traditional Chinese medicine composition has a good prevention and treatment effect on myelodysplastic syndrome and related complications thereof, can effectively inhibit proliferation of hematological tumor cells, and also can inhibit proliferation of the myelodysplastic syndrome by increasing the hemoglobin level, reducing the inflammatory cytokine level and reducing the proportion of peripheral blood T cells. Anemia, inflammation and low immune function caused by the myelodysplastic syndrome can be treated. The invention provides an effective strategy for prevention or treatment of myelodysplastic syndrome and related complications thereof, and provides a new direction for application of the traditional Chinese medicine composition.

The application provides a novel coronavirus specific T cell and a preparation method and application thereof, and belongs to the technical field of functional cells. The novel coronavirus specific T cell is enriched from peripheral blood PBMC cells, and after two rounds of in-vitro amplification, the novel coronavirus specific T cell with the order of 10 billion, an amplification multiple higher than 10,000 times and a purity higher than 90% is obtained, and the novel coronavirus specific T cell shows the characteristics of specific immune response to the novel coronavirus, thereby providing a new idea for preventing and treating the novel coronavirus pneumonia in clinic.

The application belongs to the technical field of biological medicine, and particularly relates to a TCR sequence targeting KRAS G12D antigen and application thereof. The CDR3 region of a wild type TCR is rationally designed and optimized through a simulation docking technology, and an optimized TCR with significantly improved affinity is obtained. Experimental results show that the TCR-T cell (human peripheral blood T cell TCR Homo sapiens) can effectively kill solid tumor cells, inhibit tumor growth in vivo and in vitro, improve the T cell immunotherapy effect, and can be used for preparing a cell drug for solid tumors, and has the advantages of high safety, strong targeting, significant effect and the like. The application provides a new drug selection and treatment scheme for the treatment of solid tumors.

The invention discloses an application of a Gal-9 + Th cell and / or Gal-9 + Treg as an AIH detection target spot. Researches find that compared with a healthy control group, the peripheral blood Gal-9 + Treg cell proportion and Gal-9 + Th cell proportion of an AIH patient baseline group are remarkably increased, and after ALT of a patient in a treatment group returns to normal, the Gal-9 + Treg cell proportion and the Gal-9 + Th cell proportion are both decreased compared with those before treatment, and it is indicated that the Gal-9 + Treg cell proportion and the Gal-9 + Th cell proportion are effective indexes for monitoring disease improvement. However, the AIH is a chronic disease process, even if the ALT level returns to normal, Gal-9 + Treg cells and Gal-9 + Th cells still do not return to the level of a healthy control group, the Gal-9 + Th cells are taken as detection targets, the progress of the AIH disease course can be monitored more accurately, and disease repetition and biochemical rebounding caused by premature drug withdrawal are prevented.

The invention belongs to the field of molecular targeted therapy, and discloses a small-molecule inhibitor CTPI3 of a targeted mitochondrial citric acid transporter SLC25A1 and application of the small-molecule inhibitor CTPI3 in tumor therapy. On the basis of an existing inhibitor CTPI2 structure, molecular design is carried out through an induced fit docking (IFD) module of Schrdinger software, and CTPI3 is obtained. Experiments show that the median inhibitory concentration (IC50) of CTPI3 on acute myelogenous leukemia (AML) cell lines (such as Kasumi-1 and THP1) and primary AML cells is obviously lower than that of CTPI2, the synergistic effect of CTPI3 and Venetoclax is enhanced, the mitochondrial ATP yield is reduced, and the TCA cycle metabolite level is affected. Animal experiments prove that CTPI3 can significantly prolong the lifetime of AML model mice and reduce leukemia load (spleen weight is reduced, and bone marrow and peripheral blood GFPcells are reduced), and has no significant toxicity (liver, kidney and brain histopathology is normal). The invention provides an efficient and safe new candidate drug for SLC25A1 targeted therapy.

The invention discloses a wolfberry-astragalus compound polysaccharide with an anti-myelosuppression effect as well as a preparation method and application of the wolfberry-astragalus compound polysaccharide. The wolfberry-astragalus compound polysaccharide is prepared by adopting a process of separately extracting wolfberry polysaccharide and astragalus polysaccharide and then mixing, the total sugar content is 70-80%, the uronic acid content is 10-14%, and the wolfberry-astragalus compound polysaccharide has specific molecular weight distribution and monosaccharide composition characteristics. Animal experiment results show that the wolfberry and astragalus compound polysaccharide can significantly improve peripheral hemopenia of myelosuppression mice induced by cyclophosphamide, promote recovery of the number of leukocytes, lymphocytes, monocytes and neutrophils, and effectively reverse weight loss caused by chemotherapeutic drugs. The wolfberry-astragalus compound polysaccharide provided by the invention is high in safety and remarkable in anti-myelosuppression effect, and can be used for preparing medicines or functional preparations for improving myelosuppression related diseases.

The invention belongs to the technical field of biological medicine, and discloses application of JAM3 protein in preparation of a medicine for inhibiting formation of a neutrophil extracellular trapping net. JAM3 is an I-type single transmembrane protein containing 310 amino acids, and belongs to an immunoglobulin superfamily. Researches show that the JAM3 protein remarkably inhibits excessive formation of a sepsis neutrophile granulocyte extracellular trapping net by regulating and controlling an activation path of neutrophile granulocytes, and meanwhile prevents blood platelets from being excessively captured, so that formation of microthrombus in the liver and damage to organs are avoided, disease progression is effectively controlled, and recovery of peripheral blood cells and organisms is promoted.

The invention discloses an in-vitro amplification method for improving the purity of natural killer cells. The use combination of the cell factors is IL-2, IL-15, IL-18 and IL-21, the culture of the umbilical cord blood NK cells is optimized mainly from the aspects of the factor combination used for culturing the NK cells, the use concentration and the like, CD3-CD56 + positive indexes all reach 80% or above, and the amplification rate reaches 100 times or above. The method is also suitable for large-scale culture of peripheral blood NK cells, the CD3-CD56 < + > positive rate reaches 99.0% or above, the amplification rate reaches 1000 times or above, and the killing rate of K562 cells is 30% or above when the effect-target ratio is 1: 1.

Human pluripotent stem cells (hPSCs), especially induced pluripotent stem cells (iPSCs) provide a promising starting material to produce mimetic innate immune cells such as natural killer (NK) cells and γδ T-cells for cancer immunotherapy. To facilitate consistent mass production, an overall manufacturing scheme to make mimetic innate immune cells from hPSCs was designed and demonstrated. Particularly, a robust protocol to differentiate hPSCs into NK cells or γδ T-cells through sequential hematopoietic differentiation on stromal cell line deficient in expressing M-CSF and lymphoid commitment on stromal cell line deficient in expressing M-CSF ectopically expressing DLL1 without employing CD34+ cell enrichment and spin embryoid body formation is established. Using this two-stage protocol, the generation of functional mimetic NK cells and functional mimetic γδ NKT-cells was demonstrated from hPSCs, including hESCs, peripheral blood cell-derived iPSCs (PBC-iPSCs). non-T cell-derived iPSCs or γδ T cell-derived iPSCs and the use of these mimetic innate immune cells in killing cancer cells.

The invention discloses application of a B cell depleting agent to preparation of a medicine for preventing and / or treating depression and application of a B cell detection reagent to preparation of a reagent for monitoring the depression curative effect and / or assisting in screening depression, and belongs to the technical field of biological medicine. It is found for the first time that B cell constitution of peripheral blood samples of depression patients is obviously higher than that of healthy people. Therefore, the purpose of assisting in screening depression can be achieved by detecting the B cell composition ratio of the peripheral blood sample. It is found for the first time that depression can be effectively treated by applying CD20 monoclonal antibody to a depression animal model to deplete peripheral B cells. In addition, by detecting the composition ratio of the B cells of the peripheral blood sample and quantitatively analyzing the number of the B cells of the peripheral blood, the purpose of monitoring the depression treatment effect can be achieved, and the method has good practical application value. The method can be used for researching the pathogenesis of depression and finding possible indexes for prompting prognosis, and has a wide application prospect.

The invention provides a method for in-vitro amplification and induced activation of iNKT cells, and belongs to the technical field of immune cell preparations. The in-vitro amplification method comprises the following steps: S1, separating peripheral blood PBMCs cells; s2, adding a part of PBMCs into a culture medium I for stimulation culture and irradiation, and separating to obtain irradiated PBMCs; s3, re-suspending the other part of PBMCs cells in a culture medium II, adding alpha-GalCer and IL-2 to stimulate culture, then adding the irradiated PBMCs cells, and amplifying iNKT cells; the induction and activation method comprises the following steps: amplifying the iNKT cells, adding a culture medium III, adding a dasatinib derivative, and inducing the iNKT cells to be mature. According to the method, amplification and induced activation of the iNKT cells can be rapidly promoted, the amplification efficiency is high, the obtained iNKT cells are high in cytotoxicity, the method is safe, and graft versus host reaction is not caused.

The application belongs to the technical field of biological medicine, and particularly relates to application of TREM-2 in preparation of a therapeutic drug and / or a diagnostic reagent for multiple sclerosis. The application research finds that TREM-2 is highly expressed in peripheral blood CD4+ T cells of multiple sclerosis patients, and positively correlated with the severity of the disease, indicating that TREM-2 can be used as a diagnostic marker for multiple sclerosis. The marker can be taken from peripheral blood, and has the characteristics of small trauma, low cost, rapidness and high sensitivity compared with other examinations such as cerebrospinal fluid examination, evoked potential and head CT, magnetic resonance imaging and the like. The application also finds that TREM-2 is involved in the transformation of pathogenic Th17 cells and the secretion of IL-6 and IL-17, indicating that intervention of the expression of TREM-2 can relieve and even hopefully treat multiple sclerosis.

The present invention discloses the use of methyl α-D-glucopyranoside in the preparation of thalassemia therapeutic drugs. Comprehensive analysis shows that when methyl α-D-glucopyranoside was intraperitoneally injected into β-thalassemia mice, it was found that methyl α-D-glucopyranoside was one of the metabolites with the largest down-regulation amplitude, and had multiple beneficial effects on Th3 / + mice, including increasing the total red blood cell count, hemoglobin level and hematocrit, and reducing the coefficient of variation of the red blood cell volume distribution width; reducing the iron content in the liver, splenomegaly and iron content in the spleen; increasing the GSH / GSSG ratio, reducing the ROS level, reducing the apoptosis rate of peripheral blood cells, and increasing the lifespan of red blood cells. After GP treatment, the ineffective hematopoiesis, anemia and iron overload of thalassemia mice were effectively alleviated without obvious toxic side effects. The treatment is convenient to implement, and GP is cheap and easy to store.

The invention discloses a freeze-drying preservation solution as well as a preparation method and a freeze-drying preservation method thereof. The freeze-drying preservation solution comprises the following components: trehalose, PVP-40KD, BSA (Bovine Serum Albumin), mannitol, EGCG (Epigallocatechin Gallate) and a buffer system. The freeze-drying preserving fluid provided by the invention can reduce oxidative stress reaction caused by freezing through the synergistic effect of multiple components, can effectively improve the survival rate and activity of cells after freeze drying and recovery, and can be well suitable for long-term activity preservation of peripheral blood T cells, B cells and NK cells (TBNK cells); the freeze-drying method provided by the invention can be used as a method for replacing the traditional cryopreservation, the cell preservation does not need to maintain a low-temperature environment, the cell can be transported in a room-temperature environment, the storage condition is simplified, and the long-term preservation of the cell is verified to reach 2 years by a 37 DEG C accelerated stability test.

Methods, devices, and systems for managing peripheral blood cells are provided. In one aspect, a fully automatic peripheral blood separation method includes: adding a peripheral blood sample liquid in a sample bag and a gradient liquid into a centrifuge cup, collecting a target cell liquid into an intermediate bag after centrifugation of the peripheral blood sample liquid and the gradient liquid in the centrifuge cup, adding the target cell liquid in the intermediate bag and a washing liquid to the centrifuge cup for washing and replacement to obtain a pre-product, and mixing the pre-product and a cell dilution liquid to obtain a final product.

The invention provides a biomarker screening method and system for radiotherapy-immunity pneumonia prediction, and relates to the technical field of biomedicine. Aiming at the problem that the timeliness and the specificity of the existing radiotherapy-immunity pneumonia diagnosis cannot be considered at the same time, the method comprises the following steps: separating peripheral blood T cells of a radiotherapy-immunity pneumonia group and a control group, carrying out T cell clustering annotation in combination with single-cell secretory proteomics, an R language and an ImMen database, and screening a key T cell subset Tc17 by utilizing an Augur algorithm; the method comprises the following steps: determining pericytes as key interaction cells of Tc17 through cell interaction analysis, and screening a radiotherapy specific up-regulation gene MFAP4; and carrying out difference analysis on the Mfap4hi week cell subpopulation, and screening out a biomarker gene set containing 18 genes on the basis of an elastic network algorithm in combination with 3-fold nested cross validation. The invention provides a key basis for early prediction and accurate intervention of radiotherapy-immunity pneumonia, and has the advantages of high specificity and strong repeatability.

The application relates to the field of biological medicine, and particularly discloses a monoclonal antibody combined with human CD8 and a preparation method and application thereof. The antibody is obtained by combining single B cell sorting, gene cloning and mammalian cell expression technology after rabbit immunization of a CD8 polypeptide antigen, a New Zealand white rabbit is immunized by adopting a KLH coupled CD8 antigen polypeptide combined with a rapid immunoadjuvant, high-affinity antibody clones are obtained by single B cell sorting technology, and the antibody is prepared by recombination expression. The antibody is verified by ELISA and flow cytometry, has high titer and strong specificity, can effectively recognize human peripheral blood CD8+ T cells, and has a signal strength and a background signal-to-noise ratio which are significantly better than those of a traditional mouse-derived antibody. The application provides light and heavy chain variable region amino acid sequences of the antibody, realizes controllable preparation of the antibody and protection of intellectual property rights. The antibody can be widely applied to the fields of flow cytometry detection, immunohistochemistry, tumor immunotherapy monitoring and T cell function research, and has important scientific research and clinical application values.