28 results about "Healthy donor" patented technology

The discovery and application of immune checkpoint inhibitors (ICI) have significantly promoted cancer treatment. However, the response rate is still low in gastrointestinal (GI) cancers. To explore novel adjuvant treatment options, we performed fecal microbiota transplantation (FMT) of healthy donors on 10 GI cancer patients with resistance to anti-PD-(L) 1 treatment. The combination of FMT and anti-PD1 treatment was well tolerated and no serious adverse reaction was observed. Two out of 10 patients restored clinical responsiveness to anti-PD1 after FMT. Clinical benefits were shown to be associated with better colonization of co-occurred immunogenic microbes from donors, alongside activated immune status reflected by the peripheral immune cell population. Moreover, we identified microbial signatures of ICI responsiveness and validated in an independent cohort. Our study demonstrates the feasibility of FMT as a potential adjuvant therapy for ICI irresponsive GI cancer patients and laid a foundation for LBP / consortia development to enhance ICI efficacy.

The invention belongs to the technical field of biomedicine, and particularly relates to a PAD-T cell targeting acute myelogenous leukemia cells and a preparation method and application of the PAD-T cell. The PAD-T cell comprises a T cell, a capsule shell coating the T cell, and a DNA aptamer connected with the capsule shell; wherein the capsule shell is formed by alternately overlapping a first polylysine layer, a first sodium alginate layer and a second polylysine layer, the DNA aptamer layer is an S30-T1 DNA aptamer, and the nucleotide sequence is as shown in SEQ ID NO. 1. According to the invention, the peripheral blood mononuclear cells of a healthy donor can be prepared on a large scale by utilizing a nano coating technology, and the recurrence after hematopoietic stem cell transplantation can be effectively relieved and the occurrence of GVHD can be reduced by combining an immune isolation concept and a biological material-nucleic acid material coated immune cell technology; the clinical application prospect is very high.

Provided is a technique for preparing universal DNT cells. The raw materials for preparation are derived from'white filtering residues' after white filtering treatment is carried out to avoid blood transfusion adverse reactions, and the DNT cells extracted from the white filtering residues can be a mixed culture of one or more healthy donors. The universal DNT cell obtained by using the method provided by the invention has the characteristics of high yield, high purity, high in-vitro tumor killing activity, suitability for large-scale production and the like. The initial raw material can be fresh cells or re-thawed cells after cryopreservation, the raw material source bottleneck of large-scale preparation of cell drugs is fundamentally solved, the value of leukocytes filtered out in the production process of blood products is re-excavated, and great medical value and commercial application prospects are achieved.

The present invention relates to modified T cells and methods of making and using the same. Specifically, disclosed herein are modified primary human T cells, and populations thereof, comprising a genome in which the CTLA4, PD1, TCRA, TCRB, and / or B2M genes have been edited to produce off-the-shelf universal CAR T cells from allogeneic healthy donors, the off-the-shelf universal CAR T cells can be administered to any patient while reducing or eliminating the risk of immunological rejection or graft versus host disease, and are not susceptible to T cell inhibition; and methods for allogeneic administration of the cells to reduce the likelihood that the cells will trigger a host immune response when the cells are administered to a subject in need thereof.

To provide: compositions of cells with mitochondrial DNA reduction and / or mitochondrial DNA replacement; methods for producing the same; and methods for treating various diseases associated with genetic or age-related mitochondrial dysfunction.SOLUTION: A method for producing mitochondria replaced cells includes: (a) contacting recipient cells with an agent that reduces endogenous mtDNA copy number; (b) incubating the recipient cells for a period of time sufficient for the agent to partially reduce the endogenous mtDNA copy number in the recipient cells; and (c) co-incubating (1) the recipient cells from the step (b) in which the endogenous mtDNA has been partially reduced with (2) exogenous mitochondria from a healthy donor for a period of time sufficient to non-invasively transfer the exogenous mitochondria into the recipient cells, thereby producing the mitochondria replaced cells.SELECTED DRAWING: Figure 1A

This invention relates to the application of mitochondrial formulations in the preparation of drugs for treating chronic obstructive pulmonary disease (COPD). The mitochondrial formulations contain active mitochondria isolated from healthy donor cells, with a particle size of 400–1000 nm, possessing normal mitochondrial membrane potential and ATP generation function, and capable of being internalized by mammalian COPD lung epithelial cells. In vitro cell experiments have confirmed that this mitochondrial formulation can repair CSE-induced mitochondrial dysfunction in lung epithelial cells, inhibit mPTP opening, reduce mtDNA leakage into the cytoplasm, thereby blocking the activation of the cGAS-STING signaling pathway and reducing the release of inflammatory factors such as IL-1β and TNF-α. In vivo experiments in COPD mouse models show that tail vein injection of the mitochondrial formulation can effectively accumulate in lung tissue, significantly improving lung pathological changes such as emphysema and collagen fiber deposition in model mice. This invention provides a novel strategy for COPD treatment based on repairing mitochondrial function and intervening in inflammatory signaling pathways.

The present invention involves the use of Caspase-1 inhibitors for treating ineffective erythropoiesis in patients with sickle cell disease (SCA). It highlights increased levels of IL-18 positive cells and Caspase-1 activity in hematopoietic stem cells (HSCs) of SCA patients compared to healthy donors. The invention demonstrates that treatment with a Caspase-1 inhibitor reduces this activity, suggesting its potential therapeutic benefit.

The invention discloses a microbial freeze-drying embedding method based on a pH-triggered phase change mechanism. The method comprises the following steps: step 1, preparing a zein solution: dissolving zein into a sodium hydroxide solution; step 2, preparing a bacterial suspension: taking fresh excrement of a healthy donor as a raw material, diluting with normal saline, uniformly stirring and mixing, and filtering with a multi-stage filter screen to obtain the bacterial suspension; 3, preparing an intestinal flora freeze-drying protective agent: dissolving lactose, maltotriose, mannitol, trehalose and antifreeze protein in normal saline according to a ratio, and sterilizing for later use; step 4, constructing a composite system: centrifugally washing the bacterial suspension, adding a freeze-drying protective agent, uniformly mixing in a vortex manner, adding a zein solution in proportion, and sufficiently and uniformly mixing; and step 5, freeze-drying treatment: performing vacuum freeze-drying on the composite system to form a compact hydrophobic protective layer, thereby obtaining the freeze-dried intestinal flora powder. According to the invention, the pH response characteristic of the protein is coupled with a freeze-drying process, so that efficient embedding protection of intestinal flora is realized.

The present invention relates to the prevention and treatment of disease like cancer. The inventors have previously characterized MELOE-1 antigen as an IRES dependent, melanoma specific translation product from a lncRNA mainly transcribed in the melanocytic lineage. MELOE-1 contains numerous class II epitopes and one HLA-A*0201-restricted CD8 epitope eliciting a frequent repertoire of high avidity T cells. They designed various synthetic long peptide (SLPs) comprising a CD4 epitope coupled to the CD8 epitope by a serie of linkers of 4 to 6 aa and studied the efficacy of T cell clone activation by SLP-loaded DC in vitro. Particularly, they evaluated the ability of a few selected SLPs to stimulate specific T cells proliferation of PBL from healthy donors in vitro and finally, they explored the vaccination potential of their best SLP candidate in vivo in an HLA*A0201 / HLA-DRB0101 transgenic mouse. Thus, the present invention relates a SLP comprising a CD4 class II peptide linked to a CD8 class I peptide by a specific linker and its use in the treatment of disease like cancers.

The invention provides a targeting exosome for hair regeneration and a preparation method thereof. The method comprises the following steps: separating dermal mastoid cells with a cell marker CD133 from a hair follicle strain of a healthy donor, and culturing the dermal mastoid cells in a laboratory conforming to a Taiwan GTP cell preparation factory domain in China in a heterologous and serum-free environment. Then, a culture solution containing exosomes is collected from the cell culture solution, and the multiple exosomes have surface markers such as CD9, CD63 and CD81; and finally, separating, purifying and concentrating the exosome from the culture solution by utilizing a tangential flow filtration system method or a size exclusion chromatography method and other technologies to obtain the final targeting exosome freeze-dried powder product. The invention also provides a liquid hair growth preparation prepared from the exosome product.

The invention relates to a method for rapidly preparing exosomes by using human peripheral blood. In one aspect, the method for preparing plasma exosomes from peripheral blood comprises the following steps: (1) rapidly separating plasma components of the peripheral blood obtained from healthy donors or non-specific disease patients by using a differential centrifugation method; (2) performing thermal inactivation to remove a complement system in the plasma; (3) gradually removing impurities (such as substances including protein, polypeptide, fibrinogen and the like) in the plasma by a differential centrifugation method; and (4) precipitating the exosome component in the plasma by a PEG6000 method, and washing the precipitate to obtain the exosome. The invention further relates to the plasma exosome prepared from peripheral blood, the maximum value of particle size distribution of the plasma exosome is within the range of 80-200 nm, the positive rate of CD9-PE is larger than 70%, and the positive rate of CD81-PE is larger than 80%. The method disclosed by the invention can be used for rapidly and efficiently obtaining the exosome from the human peripheral blood.

The present invention relates to methods and compositions for generating and using myoblast chimeric cells (MCCs) for treating a muscle disease, such as muscular dystrophy, where the MCCs are composed of a myoblast derived from a patient with muscle disease (MD) and a myoblast from a donor without the MD (e.g., a healthy donor). In certain embodiments, cell fusion methods are performed using 2-4, or 5, times passaged myoblasts from the MD and donor subject, and / or polyethylene glycol 0.5-1.5 g / ml. In other embodiments, the MCCs created by fusion are passaged 1-5 times before use, and are passaged at 60-80% confluency. In further embodiments, the myoblasts and / or MCCs are tested at any stage during the process for less than 5-10% CD34 and / or CD45 expression, and / or greater than 50-70% CD56 and / or CD90 expression.

The present application relates to the application of mitochondrial transplantation preparation in the preparation of a drug for treating non-alcoholic fatty liver disease, wherein the mitochondrial transplantation preparation comprises active mitochondria isolated from healthy donor cells, and has normal membrane potential and mitochondrial respiratory function. In vivo experiments on a non-alcoholic fatty liver mouse model prove that tail vein injection of the mitochondrial transplantation preparation of the present application can effectively reduce the weight of the mouse, improve the fatty liver condition of the mouse, reduce the accumulation of fat in the liver tissue, and at the same time restore the abnormal liver function of the non-alcoholic fatty liver mouse.

The application belongs to the technical field of medicine, and particularly relates to a CAR-gammadelta T cell targeting GP350 and a preparation method and application thereof. The application provides an antibody or antigen binding fragment thereof targeting GP350. The GP350-targeting antibody screened is used as an antigen binding domain to construct a chimeric antigen receptor, and a gammadelta T cell isolated from peripheral blood of a healthy donor is used to prepare a CAR-gammadelta T cell targeting GP350. In-vivo and in-vitro experiments show that the CAR-gammadelta T cell targeting GP350 has obvious killing activity on an EBV-positive nasopharyngeal carcinoma cell (GP350 + ). The application can be applied to the diagnosis of EBV-positive diseases, and has a broad prospect in the preparation of products for detecting, preventing and / or treating GP350-positive tumors.

The present invention discloses a preparation method and application of a universal CAR-T cell targeting FLT3 gene editing. The anti-FLT3 antibody or antigen-binding fragment thereof provided by the present invention includes CDRs in the heavy chain variable region, the amino acid sequence of the heavy chain variable region is shown in SEQ ID NO: 6, 12 or 17, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO: 7. The chimeric antigen receptor is constructed using the FLT3-targeting antibody or antigen-binding fragment thereof as the antigen-binding domain, and the universal CAR-T cell prepared using T cells isolated from the peripheral blood of healthy donors has significant killing activity against FLT3-positive tumors such as acute myeloid leukemia, and can be used to prepare drugs for the prevention and treatment of FLT3-positive tumors.

The invention relates to application of serum piRNA-2572505 as a breast cancer early diagnosis marker, and belongs to the technical field of medical treatment and public health. The invention firstly provides a marker for early breast cancer diagnosis, wherein the marker is serum piRNA-2572505; meanwhile, the marker is prepared into a kit for early breast cancer diagnosis. Compared with healthy donors, the expression of piRNA-2572505 in early breast cancer patients is remarkably improved, and the good early diagnosis capability is achieved; meanwhile, the expression level of the piRNA-2572505 is related to the age, so that the piRNA-2572505 can be used for screening indexes of women aged less than 45 years old.

The invention belongs to the technical field of cellular immunity, and particularly relates to a DC-CIK cell culture method based on an HBV protective antibody positive healthy donor and application, the cell culture method comprises the following steps: selecting HBsAg negative and anti-HBs antibody positive healthy donor peripheral blood mononuclear cells, loading a dendritic cell (DC) by using a mixed antigen of HBcAg and HBsAg, and culturing the DC-CIK cell by using a cell culture medium. And co-culturing the DC cells and cytokine-induced killer cells (CIK) to finally obtain the allogenic DC-CIK cells. According to the invention, DC cells of healthy donors and CIK cells are co-cultured to obtain allogenic DC-CIK cells, an HBcAg and HBsAg mixed antigen loading technology is adopted to significantly improve the antigen presentation capability of the DC cells, an optimized culture system enhances the targeting killing function of the cells, and the HBsAg clearance rate and the cccDNA inhibition rate can be effectively improved in combination with nucleoside analogue treatment, so that the DC-CIK cells can be used for preparing the DC-CIK cells. The synergistic breakthrough of efficient virus removal, immune function reconstruction and low recurrence rate in hepatitis B treatment is realized.

The present application belongs to the technical field of biological medicine, and particularly relates to an oral preparation of lipoxygenase of a healthy donor treatment type intestinal flora exosome package and a preparation method and application thereof. The oral preparation of lipoxygenase of a healthy donor treatment type intestinal flora exosome package comprises 0.18-0.78wt% lipoxygenase, 0.24-0.48wt% fucoidan, 1.0-2.0wt% tetrahydropyrimidine and a healthy donor treatment type intestinal flora exosome supplementing to 100% according to weight percentage. The present application adopts an ultrasonic method to prepare the lipoxygenase of a healthy donor treatment type intestinal flora exosome package, uses the treatment type intestinal flora exosome as a natural medicine carrier, successfully solves the technical problem of low oral bioavailability of lipoxygenase, and provides a new effective means for enhancing immunity and preventing and treating tumors.

A method for preparing targeted dermal papilla cell-derived exosomes is provided. The method involves first isolating a plurality of dermal papilla cells (DPCs) with a cell surface marker CD133 from the hair follicles of a healthy donor, in a xeno-free and serum-free environment in GTP lab in Taiwan (or GMP lab in international). Subsequently, a cell culture medium containing a plurality of exosomes is collected, wherein these exosomes possess surface markers such as CD9, CD63, and CD81. Finally, the exosomes are isolated, concentrated, and purified from the culture medium using techniques such as tangential flow filtration system (TFF system) or size exclusion chromatography, thereby obtaining the final targeted exosome lyophilized product. A liquid hair growth formulation formulated with this exosome product is also provided.

Disclosed herein is a method of diagnosing an immune-mediated or inflammatory prothrombotic condition in a subject, the method comprising: a. Combining a sample of plasma obtained from the subject with a sample of whole blood obtained from a healthy donor to obtain a combined sample, b. Contacting the combined sample with at least one agonist to obtain a test sample, c. Determining the proportion of procoagulant platelets in the test sample and in at least one control sample, said determining comprising: i. Contacting the test sample and the at least one control sample with GSAO and an alpha granule detection agent, wherein platelets in the test sample and the at least one control sample which have both increased uptake of GSAO and increased surface expression of alpha granule protein compared to an assay control sample are identified as procoagulant platelets, and d. Determining whether the subject is suffering from the immune-mediated or inflammatory prothrombotic condition, wherein an increased proportion of procoagulant platelets in the test sample compared to the at least one control sample indicates that the subject is suffering from the immune-mediated or inflammatory prothrombotic condition.

This application discloses the application of mitochondrial preparations pretreated with lipoic acid and NMN in the prevention and treatment of doxorubicin-induced myocardial injury. This application pre-treats healthy donors with lipoic acid and NMN to obtain functional myocardial mitochondria with significantly enhanced antioxidant capacity and energy metabolism efficiency. These pre-treated mitochondria are then directly injected in situ into the doxorubicin-damaged myocardium, achieving a synergistic effect of "pretreatment enhancement + local transplantation." This results in significantly better outcomes than conventional mitochondrial transplantation or systemic drug administration alone in terms of efficient colonization and functional replacement. By organically combining donor pretreatment with recipient local transplantation, this application effectively overcomes the shortcomings of existing technologies, such as poor targeting and insufficient efficacy, providing a highly efficient, targeted, and synergistic new treatment strategy for doxorubicin-induced acute myocardial injury.

Despite the notion that human CD8+ T cells are the final mediators of autoimmune β-cell destruction in type 1 diabetes (TID), none of their target epitopes has been demonstrated to be naturally processed and presented by β cells. The inventors therefore performed an epitope discovery study combining HLA Class I peptidomics and transcriptomics strategies. Inflammatory cytokines increased β-cell peptide presentation in vitro, paralleling upregulation of HLA Class I expression. Peptide sources included known β-cell antigens and several insulin granule proteins. Secretogranin V (SCG5 / 7B2) was identified as a novel β-cell antigen, which was processed into HLA-A2- and HLA-A3-restricted epitopes recognized by circulating naïve CD8+ T cells in type 1 diabetic and healthy donors. HLA-A2-bound neo-epitopes were also represented and originated from an alternative SCG5-009 mRNA splice isoform. Accordingly, the present invention relates to antigenic peptides derived from secretogranin V and uses thereof for the diagnosis and treatment of T1D.

The invention discloses a computer device, a method and a computer product for transplanting and matching intestinal flora. According to the method, the intestinal flora microecology is modeled in a GNN isograph, the intestinal flora matching method based on a healthy donor and an indication receptor is constructed on the basis of a Transform deep learning model architecture, and an analytic hierarchy process scoring algorithm is performed by combining intestinal bacteria indexes such as intestinal types and intestinal microbiome health indexes (GHMI), so that the intestinal flora microecology can be identified. The analysis accuracy of an intestinal bacteria matching model can be remarkably improved, and the method can be applied to analysis of an intestinal microbiome relationship, intestinal similarity matching, micro-ecological difference analysis in different diseases and the like.

The invention relates to the technical field of cell biology, and particularly discloses a peripheral blood source CIK cell preparation method and application thereof in tumor inhibition, the peripheral blood source CIK cell preparation method comprises the following steps: separating PBMC from healthy donor peripheral blood, performing IFN-gamma pre-stimulation amplification, transferring into a three-dimensional dynamic culture system, adding IL-2, IL-1alpha and CD3 antibodies, curcumin and resveratrol, and culturing in a three-dimensional dynamic culture system to obtain the peripheral blood source CIK cell. And knocking out a non-tumor chemokine receptor through CRISPR / Cas9 and over-expressing a tumor specific receptor, and finally harvesting the peripheral blood-derived CIK cells. Experiments show that the peripheral blood-derived CIK cells prepared by the method have higher amplification efficiency and tumor cell specific killing ability, and show a remarkable inhibition effect in tumor treatment, in addition, the technical scheme is simple and convenient to operate, controllable in cost and suitable for large-scale production, a novel efficient cell preparation is provided for tumor immunotherapy, and the technical scheme has wide application prospects. Wide clinical application prospects are realized.

The invention discloses a culture medium and a culture method for culturing human intestinal flora in vitro, every 1000g of the culture medium contains 5-20g of casein tryptone, 1-10g of beef extract, 1-10g of soybean oligosaccharide, 0.1-1g of monopotassium phosphate, 1-15g of sodium chloride, 0.5-5mg of hemin, 1-5g of cysteine hydrochloric acid, 0.5-2g of cholate, 20-100g of microcrystalline cellulose and 1-5g of agar powder, water is added to fix the volume to 1000mL, and the pH value is adjusted to 6.0-7.5. And after high-temperature steam sterilization, cooling to 40 DEG C, adding 0.5-5 / 1000 (V / V) serum, and uniformly mixing. The invention provides a culture medium and a culture method capable of culturing flora growth in human excrement twice through passage. In-vitro culture similar to intestinal flora characteristics of healthy donors is successfully realized, so that an ideal intestinal bacteria product is provided for intestinal bacteria transplantation treatment.

The application discloses a research method for tumor cell influence mechanism of monocyte remodeling based on bone marrow inflammatory microenvironment, and relates to the technical field of tumor treatment; and comprises the following steps: S1, providing bone marrow plasma from a classic myeloproliferative neoplasm patient; S2, placing primary CD14+ monocytes from a healthy donor in an in-vitro culture system containing the bone marrow plasma, inducing phenotype remodeling of the monocytes, and obtaining the remodeling monocytes. The application establishes CD163+CD206+ double-positive remodeling monocytes as a new target for MPN treatment; through comparison experiments of direct co-culture and indirect co-culture by Transwell, it is confirmed that the promotion effect of the remodeling monocytes on HEL cell proliferation strictly depends on physical contact between cells, rather than soluble secreted factors; and the application provides a theoretical basis for developing new antibody drugs or small molecule inhibitors for blocking cell-cell interaction.

The invention discloses a biomarker for early screening, postoperative recurrence early warning and prognosis evaluation of resectable non-small cell lung cancer and application of the biomarker, and belongs to the technical field of biological medicine. Polymerase chain reaction is adopted for quantitative evaluation of the peripheral blood leucocyte telomere length (LTL), compared with healthy donors, the LTL expression level of NSCLC-resected patients is remarkably reduced, the LTL expression level in early-stage patients is also remarkably reduced, and the LTL after resection is remarkably increased compared with that before resection, so that the LTL expression level is remarkably reduced. The total lifetime and disease-free lifetime of the postoperative long telomere length group are longer than those of the short telomere length group. Postoperative LTL is determined as an independent prognostic factor of OS in both univariate analysis and multivariate analysis. In addition, area under curve (AUC) analysis shows that the prediction capability of the LTL on the OS in five years after operation is high, and AUC is 0.606; and the prediction capability on the DFS is also very good, and the AUC is 0.751. The results show that LTL can be used as a promising biomarker for early screening, postoperative recurrence early warning and prognosis evaluation of resectable NSCLC.

The invention relates to a urine bacterium transplantation liquid for treating interstitial cystitis, which comprises the following components: urine bacterium precipitate obtained by centrifuging urine of a healthy donor and sterile normal saline, and the concentration of the urine bacterium precipitate in the urine bacterium transplantation liquid is 0.01-0.05 g / L. The abundance of any strain of lactobacillus, corynebacterium, prevotella, staphylococcus and streptococcus in the urine bacterium transplantation liquid is located at the first ten ranks of the abundance of all strains.