1949 results about "Somatic cell" patented technology

The invention discloses a bionic nanofiber-hydrogel composite biological scaffold as well as a preparation method and application thereof. The bionic nanofiber-hydrogel composite biological scaffold is a composite scaffold for simulating a gray-white matter partition structure of a spinal cord tissue, and comprises bionic nanofibers and a bionic hydrogel material, wherein the bionic nanofibers are arranged in an oriented manner and simulate a white matter ordered structure around the spinal cord, and the bionic hydrogel material is combined with the bionic nanofibers and simulates gray matter in the center of the spinal cord. According to the bionic nanofiber-hydrogel composite biological scaffold, the biological activity of loaded stem cells or adult cells can be improved, the problem that a large number of transplanted exogenous cells die due to unbalanced oxidative stress generated after tissue damage can be solved, and it is guaranteed that the transplanted cells reside in a damaged area and play a role; therefore, the spinal cord microenvironment is improved, proliferation and differentiation of endogenous neural stem cells are promoted, functional recovery after spinal cord injury is finally promoted, and great potential is achieved in spinal cord injury treatment.

A method and system to make a modular microfluidic device in a miniaturized form for cell culture, co-culture, and organ-on-a-microfluidic device as in-vitro model of human organs within compartmentalized 3D structures is described. The modular microfluidic device enables co-culturing of heterogeneous cell assemblies that are adjacent or sequentially connected to promote enhanced cell-cell interaction. In vivo-like flow into and / or from one cell growth chamber to adjacent cell growth chamber is maintained by micro-engineered porous barriers i.e., porous walls and membranes. These porous barriers provide a tool for mimicking the paracrine exchange between cells in the human body. The system comprises of a set of microelectrodes for real-time and long-term monitoring and quantitative measurement of the impact of stimuli on the epithelial tissue permeability. The modular microfluidic system provides a platform technology for bio-fabrication of variety of miniaturized in-vitro models for mimicking the structure and function of single and multi-organ models.

The invention discloses a nanometer antibody targeting FAP, a CAR-T cell and application of the nanometer antibody and the CAR-T cell. The invention provides nano-antibodies VHH1 and VHH2 capable of simultaneously recognizing human or mouse FAP. The nano-antibodies VHH1 and VHH2 are used as antigen binding regions to be constructed in a human or mouse CAR structure, and human or mouse FAP-CAR-T cells with a killing function can be prepared. Results of the mouse FAP-CAR-T cell or the human FAP-CAR-T cell provided by the invention show that the FAP-CAR-T cell has a good treatment effect on removal of FAP positive cells, and is a novel FAP-CAR-T cell which is expected to be applied to clinical solid tumor CAR-T therapy.

The invention relates to the technical field of biological information, in particular to an immunoreaction evaluation method based on tumor neoantigen activity sorting. The method comprises the following steps: obtaining genome data of tumor and normal tissues through whole exon sequencing, extracting multi-dimensional features of candidate somatic mutation, and screening by combining a Gaussian mixture model and a Transform model to obtain a high-confidence mutation genome set; predicting the HLA genotype of a patient based on sequencing data, translating and mutating into a peptide fragment, predicting the binding affinity of the peptide fragment and an MHC molecule by using an XGBoost model, and calculating a new antigen activity score sequence in combination with various parameters; and finally, synthesizing a new antigen peptide fragment according to a sorting result, carrying out in-vitro co-culture to detect an IFN-gamma secretion result, and dynamically optimizing a characteristic combination coefficient through a PPO algorithm to realize intelligent iterative updating, so that the accuracy of new antigen screening and the immunoreaction prediction capability are remarkably improved.

The invention relates to the technical field of carbon ion radiotherapy relative biological effects, and discloses a cell survival rate prediction method and system based on Monte Carlo simulation carbon ion spectrum distribution. The method comprises the following steps: collecting experimental data of isolated cells to construct a cell database; based on the energy range of carbon ion radiotherapy particles and the type of secondary particles, a monte carlo method is used to construct a monoenergetic particle DNA damage database. And then constructing a cell survival rate prediction model based on DNA damage repair, and fitting cell radiation specificity parameters of the model based on a cell database to accurately reflect the radiation sensitivity of the cells. Then on the basis of the monoenergetic particle DNA damage database, the mixed beam DNA damage characteristics are determined in a dose weighting mode, the mixed beam DNA damage characteristics are input into the model subjected to parameter fitting to predict the cell survival rate, the secondary particle biological effect is considered, and the prediction accuracy of the clinical carbon ion beam relative biological effect can be improved.

The present invention relates to mutant enzymes with enhanced properties and processes for oxidation of organic compound substrates using such enzymes. More specifically, provided herein are mutations in cytochrome P450 enzymes that enhance their peroxygenase activity and / or alter product selectivity compared to wild-type cytochrome P450 enzymes. Uses for such mutant cytochrome P450 enzymes are also provided, such as for biocatalytic oxidation of carbon-hydrogen bonds via a peroxygenase pathway, or for oxidizing a substrate which is an organic compound.

Fusion proteins are provided that consist of essential elements such as one or more proteins / polypeptides capable of transporting into the endoplasmic reticulum (ER) and fragments or variants of interleukin 18 (IL-18) or IL-18. The proteins / polypeptides capable of transporting into the ER include, but are not limited to, immune globular proteins (e.g., Fc regions, VHH antibodies, and scFv), and globular proteins (e.g., serum albumin). In some embodiments, the IL-18 may be a pro-IL-18 similar to a natural precursor cytokine, wherein IL-18 is fused to the C-terminus of a propeptide or a propeptide variant thereof. In some embodiments, the fusion protein may further comprise a cleavable peptide linker.

The invention discloses a bispecific cytokine fusion protein, which comprises an amino acid sequence of a type I cytokine and an amino acid sequence of a type II cytokine, and the amino acid sequence of the type I cytokine and the amino acid sequence of the type II cytokine are directly connected through a connecting peptide or are connected in series or in parallel through a fusion partner and the connecting peptide. Amino acid sequences of type I and type II cytokines are connected in series or in parallel to form the bispecific cytokine fusion protein, and the fusion protein can synergistically activate precursor depleted CD8 + T cells and terminal depleted CD8 + T cells, improve immunosuppression of a solid tumor complex microenvironment and release the potential of tumor immunotherapy. The invention further discloses nucleic acid, a carrier, a cell, a medicine or a medicine composition related to the bispecific cytokine fusion protein. The invention also discloses application of the bispecific cytokine fusion protein in preparation of drugs for treating cancers.

The invention discloses a serum-free adipogenic differentiation method for porcine adipose precursor cells, and relates to the technical field of mammalian animal cell differentiation.In the adipogenic differentiation process, an induced differentiation culture medium and a maintained differentiation culture medium are sequentially used, and the induced differentiation culture medium is composed of a basic culture medium and cell culture auxiliary factors; the cell culture auxiliary factor is composed of 66nmol / L insulin, 33 [mu] mol / L biotin, 17 [mu] mol / L pantothenic acid, 0.1 [mu] mol / L dexamethasone, 0.25 mmol / L IBMX, 1 [mu] mol / L rosiglitazone, 150 [mu] mol / L sodium oleate and 10 [mu] g / mL transferrin; the maintenance differential medium removes IBMX and rosiglitazone on the basis of inducing the differential medium. The serum-free adipogenic differentiation method for the porcine fat precursor cells, provided by the invention, is good in differentiation effect and high in repeatability.

The invention discloses a full-automatic milk somatic cell detection system and method based on fluorescence analysis. According to the full-automatic fluorescence microscopic examination system, automatic sample introduction and automatic mixing are completed through the automatic sample introduction module, automatic focusing of dyed samples is completed through the automatic focusing module, automatic photographing processing and automatic counting are completed through the automatic photographing and counting module, and therefore full-automatic fluorescence microscopic examination is achieved. According to the method, the automatic detection level is remarkably improved, the consistency of the detection process is ensured, and manual operation errors are reduced, so that the experiment efficiency, the data accuracy and the reproducibility are improved; compared with traditional off-line detection or laboratory detection, the method and system provided by the invention have better real-time performance and accuracy, and also have higher automation degree.

The invention relates to an organ-like core-shell microsphere as well as a preparation method and application thereof. The preparation method comprises the steps that 1, raw materials are prepared, specifically, shell raw materials and inner core raw materials are prepared, the shell raw materials comprise a photoinitiator and methacrylated hyaluronic acid (HAMA), and the inner core raw materials comprise organoid precursor cell suspension and matrigel; 2) molding: extruding the dispersion phase of the shell raw material wrapping the core raw material to a continuous phase by using a micro-fluidic chip and a high-precision injection pump, cutting the dispersion phase into liquid drops by the continuous phase, and performing illumination curing on the liquid drops to form core-shell microspheres; and (3) incubating, namely culturing the core-shell microspheres in a culture medium, so that the organoid precursor cells in the inner core are developed into organoids. According to the preparation method, rapid forming and curing of the organ-like precursor cells carried by the matrigel are achieved, so that the time of the organ-like precursor cells staying in the oil phase is shortened, the cells can obtain oxygen and nutrient substances, and the cell activity is improved.

The present invention relates to a method for constructing an eight-gene-edited xenogeneic organ transplantation donor pig, belonging to the field of animal biotechnology. In the wild-type porcine fetal fibroblast cell line, the GGTA1, β4GalNT2, and CMAH genes are knocked out by using the CRISPR / Cas9 gene editing technology, and the humanized genes of hCD39, hCD46, hCD55, hCD59, and hTBM are transfected. Combining with somatic cell cloning technology, GTKO / β4GalNT2KO / CMAHKO / hCD39 / hCD46 / hCD55 / hCD59 / hTBM eight-gene-edited cloned pigs are constructed. Further, through genotype, mRNA, protein expression identification and functional analysis, eight-gene-edited xenogeneic organ transplantation donor pigs are obtained. The present invention solves the technical problems of high production difficulty, low efficiency, and low survival rate of donor pigs for multi-gene-edited xenogeneic organ transplantation, and maximally solves the common problems of immune rejection reaction and complement dysregulation faced during xenogeneic organ transplantation, laying a foundation for more targeted development of donor pigs suitable for different tissue and organ xenotransplantation, and having important value for promoting the clinical transformation of xenogeneic organ transplantation.

Disclosed are compositions and methods for preventing graft versus host disease (GVHD) in subjects receiving donor cells. In particular, chimeric antigen receptor (CAR) polypeptides are disclosed that can be used with adoptive cell transfer suppress alloreactive donor cells. Also disclosed are immune effector cells, such as T cells or Natural Killer (NK) cells, that are engineered to express these CARs. Therefore, also disclosed are methods of suppressing alloreactive donor cells in a subject receiving transplant donor cells that involves adoptive transfer of the disclosed immune effector cells engineered to express the disclosed CARs.

The present disclosure provides compositions and methods useful in the treatment of trinucleotide repeat disorders, including Huntington's disease and Friedreich's ataxia. The present disclosure also provides gRNAs designed to target the HTT or FXN genes. Complexes comprising a base editor and any of the gRNAs disclosed herein are also provided by the present disclosure. The present disclosure further provides polynucleotides, vectors, cells, compositions, and kits. Methods of treating Huntington's disease and Friedreich's ataxia are also provided herein.

The invention belongs to the technical field of gene engineering, and particularly relates to PDRN, a gene engineering vector, a cell, a preparation method and application thereof. The polydeoxyribonucleotide (PDRN) provided by the invention comprises a nucleotide sequence as shown in SEQ ID NO. 1; 1 or a partial fragment containing a nucleotide sequence shown in SEQ ID NO. 1; 1 or a sequence having 80% or more of homology with the nucleotide sequence as shown in SEQ ID NO. 1. The PDRN provided by the invention can effectively reduce and inhibit the expression level of inflammatory factors TNF-alpha and IL-6 and increase the wound healing rate of HUVEC cells, and is concentration-dependent. The PDRN provided by the invention has a good application prospect in preparation of anti-inflammatory or healing-promoting products.

The invention provides a cattle FTO gene promoter mutation site and application thereof, and belongs to the field of biotechnology and genetic engineering, the mutation site is located in an FTO gene promoter region on a cattle genome 18 # chromosome, and the mutation site is specifically characterized in that an A base at a transcriptional start site upstream-33 base position is mutated into a G base (chromosome coordinate: Chr: 22040490); according to the mutation, the expression level of the FTO gene is remarkably improved, and the differentiation capacity of intramuscular fat precursor cells is remarkably enhanced; the FTO gene promoter mutation site provided by the invention not only can increase the gene expression quantity, but also can promote the differentiation of intramuscular fat precursor cells, and provides a solid technical basis for the cultivation of new beef cattle varieties and the increase of intramuscular fat content.

The invention relates to a method of modulating the expression of an endogenous gene in a somatic cell derived from an induced pluripotent stem cell (iPSC), comprising differentiating an engineered iPSC capable of stable or inducible expression of a CRISPR-related protein into a somatic cell prior to introduction of a guide RNA (gRNA) sequence. The invention also relates to the engineered iPSC, cells obtained by the method and uses thereof.

The invention discloses a somatic cell structure variation detection method based on a monomolecular long-read length sequence chart hybrid model, which comprises the following steps of: extracting a read length local sequence of a target structure variation interval and a flank region thereof, constructing a local chart genome by utilizing a partial sequence alignment chart algorithm, and encoding a multi-sequence alignment result. And then, a multi-category hybrid model is combined with an expectation maximization algorithm, and clustering analysis is performed on the comparison data, so that the somatic cell structure variation category only from the tumor sample is identified. And finally, generating a consensus sequence of a somatic cell structure variation region in the tumor sample, and taking the consensus sequence as an optimization result of a local genome. Compared with an existing short-read-length sequencing method, the method has the advantages that the problems of comparison errors, low-complexity sequences and genome heterogeneity can be effectively solved, and the detection precision of somatic cell structure variation is remarkably improved. Through the application of a single-molecule long-read-length sequencing technology, more comprehensive and accurate genome information can be provided.

Disclosed are compositions comprising a nucleic acid molecule. The nucleic acid molecule may encode an exonic sequence or portion thereof of a target ribonucleic acid (RNA) sequence. The nucleic acid molecule may further encode one or more stabilization domains. The one or more stabilization domains may be configured to reduce a cellular nuclease activity compared to a nucleic acid molecule that does not comprise the one or more stabilization domains.

The invention relates to the technical field of separation and purification, and discloses a method for extracting, separating and purifying serrapeptase in fermentation liquor through two-step salting-out. The serrapeptase in the fermentation liquor is separated and purified through a two-step salting-out extraction technology, specifically, the serrapeptase in the fermentation liquor is concentrated in a lower phase through one-step salting-out extraction, bacterial cell fragments are removed, then the serrapeptase is concentrated and purified through two-step salting-out extraction, and the serrapeptase is enriched in a middle phase. The method has good separation and purification effects, after two steps of salting-out extraction, the purification multiple of the serrapeptase is 3.5 times, the recovery rate is 90.7%, the electrophoresis purity of the obtained serrapeptase is close to 100% through SDS-PAGE gray analysis, and the serrapeptase freeze-dried powder can keep the enzyme activity of 88.6% after being stored for 180 days at the temperature of-80 DEG C.

The invention discloses a novel coronavirus neutralizing antibody, a detection kit and application of the novel coronavirus neutralizing antibody. The amino acid sequence of a heavy chain variable region of the neutralizing antibody is shown as SEQ ID No.1, and the amino acid sequence of a light chain variable region of the neutralizing antibody is shown as SEQ ID No.2. The antibody with mature affinity is screened through bioinformatics analysis of a single B cell, the antibody screening process is optimized by combining single cell RNA sequencing, VDJ rearrangement analysis and somatic cell hypermutation research, blindness of a traditional method is avoided, and the accuracy and effectiveness of antibody screening are improved. According to the neutralizing antibody GR12 provided by the invention, a heavy chain variable region and a light chain variable region of the neutralizing antibody GR12 can be specifically combined with an RBD structural domain of the SARS-CoV-2 and an S-Trimer structural domain of an Omicro variant, so that a broad-spectrum neutralizing effect on the SARS-CoV-2 virus and the variant thereof is realized. The binding activity of the antibody GR12 to S-Trimer and RBD under 2-fold and 300-fold dilution conditions is obviously superior to that of other antibodies, which indicates that the antibody GR12 has high affinity and dilution stability and is suitable for clinical large-dose administration.

Genetically modified cells that are compatible with multiple subjects, e.g., universal donor cells, and methods of generating said genetic modified cells are provided herein. The universal donor cells comprise at least one genetic modification within or near at least one gene that encodes a survival factor, wherein the genetic modification comprises an insertion of a polynucleotide encoding a tolerogenic factor. The universal donor cells may further comprise at least one genetic modification within or near a gene that encodes one or more MHC-I or MHC-II human leukocyte antigens or a component or a transcriptional regulator of a MHC-I or MHC-II complex, wherein said genetic modification comprises an insertion of a polynucleotide encoding a second tolerogenic factor.

The invention relates to a plasma metabolite composition for auxiliary diagnosis of sub-clinical mastitis of dairy cows and application of the plasma metabolite composition. The plasma metabolite combination is a first combination and / or a second combination, and the first combination is composed of glyceryl monolinoleate, 2-methylthiophene, L-cystine, (S)-(+)-citramalic acid, 9-hydroxy-10, 12-octadecadienoic acid, 16-hydroxyhexadecanoic acid, tyrosine hydroxyproline, stearoyl serotonin and histamine; the first composition is composed of 4, 5-dimethyl-2, 4, 5-trimethyloxazole, 12, 13-epoxy octadecadienoic acid, 2-cyclotetradecene-1-ketone, 4 [alpha]-carboxyl-5 [alpha]-cholest-8-ene-3 [beta]-alcohol, N-arachidonyl dopamine d8, 9 (Z) and 11 (E)-conjugated linoleic acid, and the second composition is composed of 2, 5-dihydro-2, 4, 5-trimethyloxazole, 12, 13-epoxy octadecadienoic acid, 2-cyclotetradecene-1-ketone, 4 [alpha]-carboxyl-5 [alpha]-cholest-8-ene-3 [beta]-alcohol, N-arachidonyl dopamine A product prepared based on the plasma metabolite composition and used for auxiliary diagnosis of cow subclinical mastitis is simple in use method and high in accuracy, can be used for assisting a milk somatic cell number method to diagnose cow subclinical mastitis, and reduces the probability of false negative and false positive.

The invention provides a cell development process dynamic modeling method and device based on time sequence single cell transcriptome data and a medium, and relates to the crossing field of bioinformatics and computational biology. The method comprises the following steps: constructing a Shenchang differential equation learning framework; adjusting parameters of the single cell development state change model based on the Shenxuan differential equation learning framework so as to construct a population cell development state change model; obtaining a cell specific gene regulation network and a population cell gene regulation network based on the population cell development state change model so as to predict occurrence opportunity of cell lineage differentiation and a molecular decision mechanism of cell differentiation; therefore, the problems of incomplete modeling mechanism, insufficient noise processing and lack of energy principle in the existing cell development process are solved.

Disclosed are methods of augmenting efficacy of oncology therapies by providing reprogrammed cells such as personalized regenerative cells that modulate the tumor microenvironment. In one embodiment, somatic cells are dedifferentiated into OCT-4 expressing cells and endowed with tumor homing properties through culture or gene engineering. Once a stable population of tumor homing cells is established, such cells are made to express immune stimulating agents constitutively, or selectively upon entering the cancer microenvironment. Selective expression of immune stimulatory agents can be induced by exposure to hypoxia, acidosis, immune suppressive signaling or inflammatory signaling.

The invention belongs to the technical field of biomedicine, and particularly relates to tumor neoantigen polypeptide based on EGFR gene mutation and application of the tumor neoantigen polypeptide. According to the effective method for predicting and screening the tumor neoantigen by using the tumor neoantigen prediction platform, the specific mutation of tumor tissues is obtained, and the neoantigen polypeptide with high immunogenicity, high affinity and high activation effect is more accurately predicted; the screened new antigen polypeptide makes up the blank of individualized antigen peptide in treatment of lung cancer NSCLC patients with EGFR-L858R somatic mutation, can be synthesized in a large scale, is used for subsequent standardized and individualized immunotherapy of EGFR-L858R mutation tumor patients, and has a good clinical application prospect.

The invention belongs to the field of biomedical materials, and particularly relates to a PBAE-DOPE-PEG2000 polymer-lipid nanocomposite and a preparation method and application thereof.The preparation method comprises the steps that a dimethyl sulfoxide-ethanol solution formed by mixing DOPE-PEG2000 lipid and PBAE serves as an oil phase, a sodium acetate / acetic acid buffer solution serves as a water phase, and the PBAE-DOPE-PEG2000 polymer-lipid nanocomposite is prepared through a microfluidic synthesis mode; the PBAE-DOPE-PEG2000 polymer-lipid nano composite material is prepared by the steps of carrying out ultrasonic treatment on the PBAE-DOPE-PEG The polymer-lipid nano composite material can efficiently load exogenous genes, the surface performance of the polymer-lipid nano composite material is changed and the stability of nano particles is improved by introducing DOPE-PEG2000 lipid, membrane fusion can be promoted, the risk of lysosome degradation is reduced, the transfection process of the exogenous genes in recipient cells is accelerated, and efficient expression in 293FT cells is realized. The efficient transfection of the polymer-lipid nanocomposite material is expected to show important application prospects in immune cell therapy or tumor cell therapy.

The invention relates to the technical field of raw milk detection, and discloses an automatic detection device for a raw milk sample, and the automatic detection device comprises a mechanical arm mechanism, an information acquisition mechanism, an incubation mechanism, a detection mechanism and a control mechanism. According to the embodiment of the invention, by arranging the mechanical arm mechanism, the mechanical arm mechanism can be used for automatically taking the sample, so that manual operation is not needed. By arranging the incubation mechanism, batch incubation and batch cleaning of the sample cups can be carried out, ultrasonic treatment can be carried out, the incubation rate and the cleaning rate are increased, and the cleanliness of the sample cups can be improved. Furthermore, according to the embodiment of the invention, the detection mechanism is arranged, so that the samples can be detected in batches, various items such as a raw milk viscosity item, a raw milk bacterial colony total number item and a raw milk somatic cell item can be detected at the same time, and compared with the current manual single item detection, the detection efficiency can be obviously improved.

The invention provides a plug-and-play Mi3 protein nanocage fusion protein with a VNP6 tag and a preparation method and application of the plug-and-play Mi3 protein nanocage fusion protein, the fusion protein comprises the VNP6 tag, a SpyCatch003 component, a Mi3 protein nanocage and a functional protein sequence, and the VNP6 tag is located at the N end of the fusion protein. The VNP6 peptide sequence or the variant thereof is utilized to induce a vesicle or vesicle-like structure beneficial to protein folding and stabilization in escherichia coli, so that the correct folding rate and activity of recombinant protein are improved while the overall intracellular molecular crowding effect is relieved. The core idea of the invention lies in that the internal structure of cells is regulated at low temperature, and the internal microenvironment of an escherichia coli expression system is optimized, so that protein molecules under high expression load can obtain more reasonable spatial distribution, and the problems of misfolding and aggregation caused by molecular crowding are reduced.

Methods, systems, and software for estimating circulating tumor fraction are provided. A first plurality of nucleic acid sequences for a plurality of loci in genomic DNA from a solid tumor sample is obtained. A second plurality of nucleic acid sequences for a plurality of cell-free DNA fragments obtained from a liquid biopsy sample from the same subject is obtained. One or more somatic mutations is identified in the first plurality of nucleic acid sequences. A variant allele frequency (VAF) is determined for each somatic mutation based on a frequency of the respective somatic mutation in the liquid biopsy sample and a frequency of the corresponding wild type allele in the liquid biopsy sample, thereby determining a set of VAFs. An estimate of the circulating tumor fraction for the test subject is determined based on the set of VAFs for the one or more somatic mutations.