1821 results about "Living cell" patented technology

The invention belongs to the field of molecular biology and immunology, and particularly relates to an anti-CD3 nano antibody capable of activating the activity of T cells as well as a preparation method and application of the anti-CD3 nano antibody. The invention provides a preparation method of the anti-CD3 nano antibody. The preparation method comprises the following steps: constructing a CD3 plasmid and an overexpression cell line, and immunizing alpaca to obtain peripheral blood mononuclear cells (PBMC) of the alpaca; extracting RNA (Ribonucleic Acid) and carrying out reverse transcription to obtain cDNA; carrying out PCR amplification on the antibody gene segment and cloning the antibody gene segment to a phage display vector; and electrically transforming the vector into SS320 competent cells to obtain the nano antibody library. After two rounds of panning, detecting and screening an antibody clone capable of being combined with the CD3 protein through ELISA (Enzyme-Linked Immunosorbent Assay); an antibody sequence is selected through sequencing, and an antibody protein is expressed by using a mammalian cell high-throughput expression system; and then detecting and screening the antibody which is only specifically combined with the CD3. The method disclosed by the invention is simple, convenient and rapid to operate, and can be used for preparing active antibodies for treating potential tumors and proliferative diseases.

A method and apparatus are disclosed. In an example from the perspective of a User Equipment (UE), the UE receives an indication of activation of Cell Discontinuous Transmission (DTX), wherein the Cell DTX is associated with one or more active periods and one or more non-active periods, and the UE is configured with a plurality of Channel State Information Reference Signal (CSI-RS) resources for deriving a CSI report. The UE determines whether to transmit the CSI report based on whether the UE receives a CSI-RS in an active period of the one or more active periods associated with the Cell DTX.

The invention provides a single particle diffusion quantitative feature prediction method and device, electronic equipment and a storage medium. The method comprises the following steps: acquiring a target input image corresponding to a target single particle; based on a pre-trained particle trajectory prediction model, according to the target input image, predicting to obtain a particle pseudo trajectory image; on the basis of the particle pseudo-trajectory image, diffusion quantitative characteristics of the target single particle are calculated; wherein the particle trajectory prediction model is obtained by performing training optimization according to a first training sample set formed by the single-particle motion blurred image and the corresponding particle motion trajectory image. According to the method, the actual movement path of the target single particle is predicted from the target input image through the particle trajectory prediction model, and the diffusion quantitative characteristic of the target single particle is calculated based on the actual movement path, so that the diffusion coefficient of a large number of single particles can be rapidly and accurately predicted in a high-mark-density environment, and the phototoxicity is remarkably reduced; diffusion characteristics of different biological particles can be effectively distinguished, and a powerful tool is provided for dynamic research of particles in a diffusion system and living cells.

The invention relates to a barrier repair composition for activating cell energy and application thereof, the composition comprises the following components: nicotinamide, a wheat germ extract, oat beta-glucan and a golden camellia flower extract, the four components synergistically form a full-link barrier repair system of energy supply (nicotinamide)-autophagy cleaning (wheat germ extract)-structure strengthening (beta-glucan)-oxidation defense (camellia nitidissima extract), the limitation that a traditional skin care product singly supplements lipid or resists oxidation is broken through, lipid dynamic balance is rebuilt from the root of cell metabolism, and the skin care product has the advantages that the skin care effect is good; and certain skin smoothing and anti-aging effects are achieved.

The invention provides a 3D biological printing osteochondral organ based on dynamic mechanical regulation and control, and a construction method and application thereof. Compared with the prior art, the invention innovatively provides a biphasic scaffold optimization modification strategy: firstly, an osteochondral hydrogel scaffold with an accurate layered structure is prepared by adopting a 3D biological printing technology, then a cell mechanical transduction signal channel is activated by a dynamic pressure stimulation system, and cartilage / bone biphasic differentiation of stem cells is directionally regulated and controlled; and by constructing a bionic mechanical microenvironment, the obtained osteochondral organ not only keeps excellent biocompatibility, but also has gradient mechanical properties and a clear spatial layered structure (cartilage layer-bone layer), the directional cartilage formation / osteogenic differentiation efficiency of the osteochondral organ is remarkably improved, and the osteochondral organ has a good application prospect. The problems of spatial heterogeneity construction and accurate regulation and control of cell fate in osteochondral tissue engineering are effectively solved, and the method has important application value in the fields of osteochondral defect repair treatment and mechanical stimulation mechanism research.

A three-dimensional scaffold for use in engineered tissue includes a repeated 3D geometry; and at least one of a hydrogel, a biocompatible polymer, and a bioprintable polymer. The repeated 3D geometry may include an open geometry including a plurality of interconnected nodes configured to be seeded with living cells. The scaffold may be formed via bioprinting. The three-dimensional scaffold may also be used for (1) 3D tissue modelling (for liver, kidney, gut, pancreas, heart, lung, spleen, etc.), (2) disease modeling of different tumor types such as breast, colorectal, lung, liver, brain, ovarian and others, (3) drug testing / therapeutic screening of small molecule drugs, peptides, proteins, antibodies, ADCs, gene therapies, RNA-based therapeutics, cell therapies, and tissue therapies, and (4) therapeutic / drug delivery (i.e., via delivery device that can be used to control drug release based on different patterns).

The invention provides an enhancer RNA (Ribonucleic Acid) molecule MZGAe1 and application thereof, and relates to the technical field of biology. The invention provides an enhancer RNA (Ribonucleic Acid) molecule MZGAe1. The nucleotide sequence of the enhancer RNA molecule MZGAe1 is as shown in SEQ ID NO: 1; meanwhile, the invention further provides a specific sgRNA sequence for activating the molecule in a targeted manner and a recombinant vector of the specific sgRNA sequence. Expression of endogenous MZGAe1 of cells is specifically activated by adopting a CRISPR activation technology, and conversion of mouse embryonic stem cells to bicellular-like cells can be efficiently promoted. By providing a brand-new endogenous targeted MZGAe1 accurate activation tool, the efficient transformation of wild mouse embryonic stem cells to bicellular cells can be realized only under the condition of endogenous activation of single enhancer RNA molecule MZGAe1, the proportion is at least increased by 5%, the expression of a totipotent marker gene ZSCAN4 is activated, and the expression of the totipotent marker gene ZSCAN4 is promoted. And an efficient and specific brand-new research tool and scheme are provided for researching zygote genome activation, embryonic development early events and cell reprogramming.

A spatiotemporal illumination microscope system is presented. The system includes a multi-well plate configured to receive live cells and a DMD configured to deliver targeted illumination to selectively illuminate a plurality of the wells. The system further includes a tandem-lens system having an objective lens and an imaging lens positioned facing each other to capture fluorescent light emitted from the cells. The system also includes an image sensor that receives fluorescent light captured by the tandem-lens system, generating imaging data. The system further includes a controller configured to use the imaging data to control closed-loop modulation of the live cells.

The invention discloses a method for improving the axial resolution of photoacoustic microscopic imaging obtained based on a single-pixel frequency domain. The method comprises the following steps: 1) creating a Fourier substrate pattern with the spatial frequency of k in the axial direction; 2) respectively irradiating four Fourier substrate patterns with the spatial frequency of k and the initial phases of # imgabs0 #, # imgabs1 #, # imgabs2 # and # imgabs3 # to an imaging object, carrying out photoacoustic microscopic imaging, and acquiring data by using an ultrasonic transducer to obtain a response value of the ultrasonic transducer; 3) according to the obtained response value of the ultrasonic transducer, adopting a four-step phase shift method to obtain a Fourier coefficient corresponding to the spatial frequency k; and 4) gradually increasing the spatial frequency of the Fourier substrate pattern to the cut-off frequency of the objective lens, further obtaining Fourier coefficients corresponding to all spatial frequencies, obtaining a complete Fourier spectrum, and performing inverse Fourier transform to obtain a photoacoustic microscopic image after the axial resolution is increased. The method disclosed by the invention is expected to promote wider application of photoacoustic microscopic imaging in living cell and tissue imaging.

The invention discloses a beta-lactoglobulin fluorescent probe as well as a preparation method and application thereof, the structural formula of the probe is (C30H25N4I), the probe is combined with beta-lactoglobulin through non-covalent interaction, a fluorescence signal is triggered to be remarkably enhanced, and low-background fluorescence detection of the beta-lactoglobulin is realized. The probe has low cytotoxicity and excellent mitochondrial targeting ability, and provides a powerful tool for visualization of beta-lactoglobulin in living cells. After being combined with beta-lactoglobulin, a sample can generate obvious color change under the irradiation of an ultraviolet lamp, and the change can be used for evaluating the content of active beta-lactoglobulin in the sample, so that the rapid detection of the milk protein level in a beverage is realized. The beta-lactoglobulin fluorescent probe disclosed by the invention has a wide application prospect in the fields of biological imaging, milk protein detection, beverage quality control and the like.

The invention belongs to the technical field of gene engineering, and particularly relates to an RNA polymerase beta subunit mutant and application thereof. The RpoB mutant is obtained through a gene editing technology, after the 560th amino acid of the RpoB is changed into leucine from proline, the number of living cells during strain fermentation is greatly increased, the tolerance of the RpoB mutant in the later environment of a fermentation tank is improved, the RpoB mutant is applied to escherichia coli for producing human milk oligosaccharide, and finally the yield of the human milk oligosaccharide is greatly increased.

Small molecule therapeutics can concentrate in distinct intracellular environments, some bounded by membranes, and others that may be formed by membrane-less biomolecular condensates. The chemical environments within biomolecular condensates have been proposed to differ from those outside these bodies, but the internal chemical environments of diverse condensates have yet to be explored. Here we use small molecule probes to demonstrate that condensates formed in vitro with the scaffold proteins of different biomolecular condensates harbor distinct chemical solvating properties. The chemical rules that govern selective partitioning in condensates, which we term condensate chemical grammar, can be ascertained by deep learning, allowing efficient prediction of the partitioning behavior of small molecules. The rules learned from in vitro condensates were adequate to predict the partitioning of small molecules into nucleolar condensates in living cells. Different biomolecular condensates harbor distinct chemical environments, that the chemical grammar of condensates can be ascertained by machine learning.

The invention relates to a culture method for activating NKT cells by DC cells in vitro and application of the NKT cells. The preparation method of the NKT cell comprises the following steps: activating a DC cell by using cell inducing factors such as alpha-GalCer, GM-CSF and IL-4, co-culturing the DC cell and a cryopreserved and resuscitated suspension cell left after adherent sorting, and activating and amplifying the NKT cell. The proportion of NKT (CD3 + CD56 +) prepared by the invention is more than 80%, and a small amount of iNKT cells are contained at the same time.

The invention belongs to the technical field of living cell imaging, and discloses a living cell nucleic acid imaging system and method based on CRISPR (clustered regularly interspaced short palindromic repeats) and a nano antibody and application, the system comprises: a modified dCas protein expression vector containing n antigen polypeptide tags; the modified sgRNA and / or crRNA expression vector comprises a specific guide sequence; the fluorescent nano-antibody expression vector contains m fluorescent nano-antibody coding sequences which are connected in an operable manner, and fluorescent protein serving as an internal tag is inserted between nano-antibody splitting sites, can be combined with a polypeptide tag carried by dCas protein and can stably emit light; and the fluorescent nano antibody in a free state can be self-degraded, so that the fluorescent nano antibody does not emit fluorescence. According to the invention, the single-copy nucleic acid sequence, especially the imaging marker of the single-copy nucleic acid sequence in a living cell, can be visualized with single-molecule resolution and high signal-to-noise ratio.

Fusion proteins are provided according to aspects of the present disclosure which include: a glycan binding component linked to a mutant E. coli biotin ligase BirA, the glycan binding component capable of specific binding to a glycosylation post-translational modification of a target protein and the mutant E. coli biotin ligase BirA having enzymatic activity to ligate biotin to proteins proximal to the target protein. Methods of detecting proteins proximal to a target protein are provided according to aspects of the present disclosure which include contacting a living cell with the fusion protein according to aspects of the present disclosure under compatible biological conditions; and detecting the biotinylated proteins, thereby detecting proteins proximal to the target protein.

The invention relates to the technical field of fluorescent dyes, in particular to a citric acid-based fluorescent dye for distinguishing dead cells from living cells as well as a preparation method and application of the citric acid-based fluorescent dye. The invention discloses a citric acid-based fluorescent dye for distinguishing dead cells from living cells. The citric acid-based fluorescent dye comprises at least one of a compound 1 and a compound 2, the citric acid-based fluorescent dye can specifically enter dead cells and specifically dye cytoplasm of the dead cells, so that the dead cells can be dyed or imaged. The citric acid-based fluorescent dye is difficult to dye living cells, so that dead cells and living cells can be quickly and accurately distinguished, and the citric acid-based fluorescent dye can be applied to the field of biological medicines.

The invention discloses a cell culture and microscopic imaging integrated device which adopts an integrated design and comprises an imaging assembly, a fluorescence excitation assembly, an objective lens assembly, an automatic shading assembly, a two-dimensional displacement platform, a cell culture assembly, a bright field light source assembly and a shell, the imaging assembly, the fluorescence excitation assembly, the objective lens assembly, the automatic shading assembly and the two-dimensional displacement platform are integrated in the shell, the cell culture assembly is arranged on the two-dimensional displacement platform, and the bright field light source assembly is arranged above the shell. The device has the advantages that the bright field light source, the fluorescence excitation module, the imaging assembly and the cell culture assembly are integrated together, the device is complete in overall function, compact in structure, free of multiple sets of systems and convenient and fast to operate, the temperature and gas environment needed by cell culture are provided through the cell culture assembly, and the requirement for long-time real-time observation of living cell imaging is met; cell culture, multichannel fluorescence imaging and analysis can be completely realized through a single device, the operation process is simplified, and the working efficiency is improved.

The invention relates to the technical field of biological microscopic imaging, and discloses a deconvolution super-resolution imaging method and device based on deep learning denoising and a medium. The method comprises the following steps: acquiring a to-be-denoised image of a to-be-imaged structure marked by fluorescent molecules; de-noising the image to be de-noised by using a preset image de-noising neural network to obtain a de-noised image; a training data set for training the network is generated by utilizing the optical switch characteristics of optical switch fluorescent molecules, and the optical switch fluorescent molecules are used for marking to-be-imaged structures of samples in the training data set; the emission spectrum of the fluorescent molecules is overlapped with the emission spectrum of the fluorescent molecules of the optical switch; and carrying out deconvolution processing on the denoised image to obtain a super-resolution imaging image. According to the invention, the large-scale and high-quality true value image is obtained by using the optical switch characteristic of the optical switch fluorescent molecules, and the denoising effect of the image denoising neural network is improved; and denoising the image by using the network, and then carrying out deconvolution on the denoised image to realize large-view living cell single-frame super-resolution imaging.

The specification describes an improved serum-free animal cell culture medium, which can used for the production of a protein of interest. Ornithine, or a combination of ornithine and putrescine can be added to serum-free media or chemically defined media to improve viable cell density, to reduce cell doubling time, and to increase the production of a protein of interest.

The invention discloses a recombinant protein and a detection reagent for detecting encephalitis of an anti-GABABR antibody, and belongs to the technical field of neuroimmunology diagnos.A GABABR1 partial sequence, a GABABR2 partial sequence and a transmembrane sequence (and a fluorescent label) are adopted to construct a cell strain for stably and highly expressing the recombinant protein by transfecting CHO cells or HEK293 cells, and the recombinant protein and the detection reagent are used for detecting the encephalitis of the anti-GABABR antibody. The sensitivity and the specificity of the kit in anti-GABABR antibody encephalitis detection can be remarkably improved, the aim of detecting the antibody by using a fixed CBA method or a living cell CBA method can be achieved, and the kit is suitable for auxiliary diagnosis of autoimmune encephalitis of cerebrospinal fluid / serum.

The invention discloses a preparation method of a cell membrane modified ultrasonic response type copper / ferroptosis activated nano-carrier. The nano-carrier is used for in-vivo and in-vitro treatment of ultrasonically activated tumors. The carrier is used as an ultrasonic activation type copper / iron death activator, and breast cancer cell membrane doped lipid is coated on the outer layer of a copper-iron bimetal organic framework to form nanoparticles with a core-shell structure. The biomimetic copper / ferroptosis activated nano-carrier disclosed by the invention is mainly characterized in that (1) the in-vivo circulation time of the nano-carrier is prolonged by modification of a breast cancer cell membrane, and accumulation of the nano-carrier in breast cancer cells and tissues is improved by virtue of homologous targeting of the nano-carrier; (2) the copper-iron double-metal-doped metal organic framework has a uniform nano structure, and has the potential of effectively catalyzing Fenton / Fenton-like reaction activity and activating a copper / iron death synergistic effect in cells; and (3) depending on the sound-sensitive agent and the oxygen carrier which are loaded by lipid, the nano-carrier has an ultrasonic signal response release behavior, and can simultaneously activate a sonodynamic effect, a copper death process and an iron death process in tumor cells, so that accurate and efficient killing of tumor tissues is realized.

KLRB1 binding agents (in particular anti-KLRB1-antibodies and antigen binding portion thereof) with increased humanness and compositions thereof, as well as therapeutic methods of using the agents, e.g., for depleting cells or inhibiting cells or activating cells, (in particular, Th17, Th17.1, ex-Th17, Tc17, MAIT, iNKT, peTh2, ILC2, ILC3, NK cells, and / or neoplastic T or NK cells in vivo), for the treatment of autoimmune disease, allergic diseases, transplant rejection, hematologic malignancies, and cancer.

The invention belongs to the technical field of microorganisms and fermentation engineering, and particularly relates to application of bifidobacterium longum subsp. Infantis Y46 in the aspects of enhancing cell defense and repair and regulating metabolic homeostasis of a host. The strain is isolated from infant feces. Animal experiments prove that living cell thalli of the bifidobacterium longum subsp. Infantis Y46, which are obtained by culturing an MRS liquid culture medium added with L-cysteine hydrochloride (0.05%, v / v), can significantly enhance the oxidative stress resistance of an organism and reduce the level of active oxygen in the organism compared with a control bacterium bifidobacterium longum subsp. Infantis 15697. Transcriptomics analysis shows that the bifidobacterium longum subsp. Infantis Y46 can enhance self-protection of cells and improve physiological imbalance caused by external pressure by regulating related signal channels of an organism. The action mechanism is closely related to signaling pathways related to cell defense, metabolic homeostasis and barrier functions. The invention provides a new strain resource and an application scheme for developing functional food or medicinal preparations for enhancing the health defense function of a human body.

The present invention relates to the field of stem cell technology. The present invention discloses a method for inducing stem cells to secrete specific factors, which includes stimulating stem cells with high-frequency waves, emitting sound waves of a specific frequency to stem cells, regulating stem cells in a hypoxic environment, culturing stem cells in a hypoxic environment, stimulating stem cells with oxidative stress, and subjecting stem cells to oxidative stimulation treatment. The present invention can significantly enhance the proliferation ability of stem cells, promote the secretion of multiple active factors, and improve cell function and adaptability. As a non-invasive physical method, high-frequency wave stimulation can activate internal cell signal pathways without introducing exogenous substances, significantly improving the proliferation ability and tissue repair potential of stem cells. Hypoxic environment regulation simulates the naturally existing microenvironmental conditions in the body, greatly promoting the secretion of active factors such as vascular endothelial growth factor, basic fibroblast growth factor, and brain-derived neurotrophic factor for tissue repair and immune regulation.

The invention relates to a coumarin-based fluorescent probe as well as a preparation method and application thereof in the aspect of HClO detection, and belongs to the technical field of HClO detection. The invention discloses a coumarin-based fluorescent probe W (O-(3-((2-morpholinyl ethyl) carbamoyl)-2-oxo-2hydrogen-benzopyran-7-yl) dimethyl thiocarbamate) prepared on the basis of an intramolecular charge transfer (ICT) mechanism, and a preparation method of the coumarin-based fluorescent probe W. The novel lysosome targeting fluorescent probe contains coumarin as a fluorophore, a morpholine group as a targeting group and a dimethyl thiocarbamate group as a recognition group, is a novel lysosome targeting fluorescent probe, and has excellent performances of high selectivity, low detection limit, low cytotoxicity, high resolution and the like in the aspect of HClO detection; the method can be used for detecting the content of endogenous and exogenous hypochlorous acid in living cells.

The invention discloses a low-quality fluorescent microscopic image reconstruction method based on noise compensation guided filtering and unmatched projection deconvolution, which efficiently realizes pre-denoising of detail preservation and background noise suppression through guided filtering of noise compensation, and takes the pre-denoising as the input of an unmatched projection deconvolution method. The blurring caused by diffraction is removed, the pre-denoising step can effectively suppress noise amplification in the deconvolution process, and the whole process realizes the improvement of the signal-to-noise ratio of the image and the restoration of the resolution. Therefore, the method disclosed by the invention can be used for simply and effectively improving the quality of the image acquired under the low-light-dose imaging condition, and is beneficial to information acquisition of long-time fluorescent imaging of the living cells.

The invention belongs to the technical field of microorganisms and fermentation engineering, and particularly relates to application of bifidobacterium longum subsp. Infantis Y46 in the anti-aging aspect. The bifidobacterium longum subsp. Infantis Y46 is obtained from excrement of infants. Animal experiments prove that living cell thalli (Y46) and thermally inactivated cell thalli (HKY46) obtained by culturing the bifidobacterium longum subsp. Infantis Y46 by using an MRS liquid culture medium containing L-cysteine hydrochloride (0.05%, v / v) can significantly reduce accumulation of lipofuscin in a host, and prolong the healthy life and life of the host. Therefore, the bifidobacterium longum subsp. Infantis Y46 is a promising probiotic and a promising metagen with an anti-aging function.

The invention discloses a living cell super-resolution imaging method and system based on self-supervised learning. The method comprises the following steps: S1, obtaining a fluorescence microscopic image through a standard fluorescence microscopic system; s2, for the acquired single noise image, generating positive and negative sample pairs of self-supervised training data through an autonomously designed and optimized self-supervised strategy; s3, constructing a front denoising neural network and a rear deconvolution network, and performing network training; and S4, inputting a newly obtained noise image into the trained front denoising network to obtain a denoised image, and inputting the denoised image into the rear deconvolution network to obtain a final super-resolution reconstructed image. According to the invention, the standard fluorescence microscopic system and the self-supervised denoising processing module are combined, so that high-quality denoising and super-resolution reconstruction of the fluorescence microscopic image can be realized in a low-photon signal scene.

The invention provides a nano-drug for treating glioma as well as a preparation method and application of the nano-drug, and particularly, the nano-drug comprises a nano-carrier and a carrier drug. The nano-drug can interact with a receptor on a brain endothelial cell, enters a brain tumor microenvironment through an endocytosis way, promotes antigen presenting cell maturation, activates T cell anti-tumor immune response, and effectively inhibits tumor growth.