542 results about "Extracellular" patented technology

The invention relates to a single cell image segmentation method based on a minimum circumcircle, which solves the technical problem of how to improve the precision, robustness and adaptability of an image-based single cell segmentation method, and comprises the following steps of: firstly, obtaining an original cell image, and secondly, converting the original cell image into a gray level image; preprocessing and binarization processing are carried out on the grayscale image to obtain a binarized image, a cell edge contour image is obtained through processing, a minimum circumcircle is drawn for each contour in the cell edge contour image to obtain a cell circumcircle image, and finally, the center of the minimum circumcircle is used as the center point of a cutting area to obtain a cell edge contour image. And determining the boundary of the cutting area by taking the background including the cells and within a certain range around the cells as a standard, cutting the cell circumcircle image, and finally obtaining a single cell image. The method is suitable for single cell identification and segmentation in a microscope image, and can be widely applied to the fields of cell biology research, medical diagnosis, drug screening and the like.

The invention discloses a multi-effect repair type collagen and application thereof, and belongs to the technical field of synthetic biology. Transdermal peptide TD-1, a transmembrane region of human XVII type collagen, an extracellular sixteenth non-collagen region, an extracellular fifteenth collagen region and a coding gene of a His tag are selected to be connected in series, and a gene sequence is optimized through pichia pastoris codon selection preference; and then the T-COL17R3 is constructed and expressed to obtain the T-COL17R3. Efficacy experiments prove that compared with similar products sold in the market, T-COL17R3 not only has excellent transdermal performance, but also has better cell proliferation and migration promoting capability, better capability of resisting early saccharification product ketoamine and later saccharification product dicarbonyl compounds, better capability of scavenging free radicals and resisting elastase, and can be used for preparing the anti-epidermal drug. In addition, synthesis of melanin in B16-F10 cells can be reduced, so that T-COL17R3 has excellent application potential in development of pharmaceutical compositions or skin care products with repairing, anti-wrinkle and firming or whitening and brightening effects.

The invention discloses a light-driven synthetic flora and application thereof. The light-driven synthetic flora comprises phototrophic bacteria and heterotrophic bacteria, the phototrophic bacteria are genetically modified synechococcus slender, and the gene modification comprises construction and expression of heterologous sucrose permease, glyceraldehyde dehydrogenase and sucrose synthase in the synechococcus slender; and the heterotrophic bacteria are the P.adami. According to the light-driven synthetic flora disclosed by the invention, carbon dioxide is converted into cane sugar through photosynthesis of phototrophic bacteria, and the cane sugar is secreted out of cells to provide a carbon source for heterotrophic bacteria; carrying out cell growth and polyhydroxyalkanoate synthesis on the heterotrophic bacteria by using cane sugar; therefore, the light-driven synthetic flora provided by the invention can realize direct conversion of carbon dioxide into polyhydroxyalkanoate without additional carbon sources, has the advantage of environmental friendliness, and provides a new economical and feasible scheme and approach for production of polyhydroxyalkanoate.

The invention relates to an antibody endocytosis efficiency detection method based on fluorescence resonance energy transfer, and belongs to the technical field of antibody endocytosis detection. The method comprises the following steps: coupling succinimide ester with an amino-containing compound to obtain a quenching agent, incubating an antibody to be detected and a target cell together, adding a fluorescence-labeled secondary antibody to carry out endocytosis reaction, removing an extracellular fluorescence signal by using the quenching agent after the reaction is finished, and detecting an intracellular fluorescence signal to obtain the endocytosis percentage of the antibody to be detected. The detection method provided by the invention is based on a fluorescence resonance energy transfer principle, an intracellular signal direct detection method in the prior art is optimized, and an extracellular fluorescence signal is removed, so that a detection system has higher sensitivity. The quenching agent provided by the invention can also be combined with an acid eluent in the prior art to further reduce an extracellular signal background value and improve the sensitivity of a detection system, can be applied to high-throughput detection of a to-be-detected antibody, is high in sensitivity and low in detection cost, and has a good application prospect.

The invention relates to the technical field of biomedicine, in particular to stem cell exosome-loaded bi-crosslinked hydrogel as well as preparation and application thereof, and the bi-crosslinked hydrogel comprises the following components in parts by weight: 50-90 parts of a matrix polymer; 0.1 to 5 parts of an active load; 1 to 10 parts of cyclodextrin; 5 to 20 parts of modified chitosan; 0.01 to 1 part of a photoinitiator; and 0.01 to 0.5 part of growth factors. Through double-network design of physical / chemical primary cross-linking and photo-initiation secondary cross-linking, the bottleneck that the mechanical property and the machinability of the traditional single cross-linking hydrogel are difficult to consider at the same time is overcome; meanwhile, the human umbilical vein endothelial cell exosome is introduced, and efficient loading, uniform dispersion and degradation coupling slow release of the exosome are realized by virtue of beta-cyclodextrin inclusion and a double-crosslinking network. Therefore, the problems of mechanics, degradation, cross-linking speed or tissue adhesion of cross-linked hydrogel in the prior art are solved.

Compositions of retroviruses and methods of using the same for gene delivery, wherein the retroviruses comprise a viral envelope protein comprising at least one mutation that diminishes its native function, a non-viral membrane-bound protein comprising a membrane-bound domain and an extracellular targeting domain.

Provided herein are novel stabiligases for use in cell membrane proteome analysis. The subject stabiligases are capable of attaching to glycans found on the surface of cell membranes to form glycan-tethered (GT) stabiligases. Such glycan-tethered stabiligases are capable of robustly and selectively attaching label probes to cell surface proteins of intact cells. The subject novel stabiligases described herein advantageously allow for the identification and profiling of cell membrane proteins that have undergone an extracellular N-terminal proteolytic event.

The invention discloses an MUC1-CAR-T cell for co-expressing a BTLA / IL-18R chimeric receptor as well as a preparation method and application of the MUC1-CAR-T cell, and belongs to the technical field of biological medicines. The MUC1-CAR-T cell comprises a BTLA / IL-18R chimeric receptor and an MUC1-CAR structure, wherein the BTLA / IL-18R chimeric receptor is a BTLA MUC1 is used as a targeting molecule for CAR-T cell therapy, an extracellular domain of BTLA is connected with a transmembrane domain and an intracellular domain of an IL-18 receptor, and the co-expression BTLA / IL-18R chimeric receptor is designed. Experimental verification shows that the BTLA / IL-18R chimeric receptor can significantly improve the tumor killing ability of MUC1-CAR-T cells, can better inhibit the growth of tumors, and provides a novel immunotherapy strategy for the treatment of pancreatic cancer.

In various aspects, the invention provides culture platforms, cell culture media, and methods of differentiating pluripotent cells into hematopoietic cells. In certain aspects, the invention also provides pluripotent stem cell-derived hematopoietic cells generated using the media and methods disclosed herein. The pluripotent stem cell-derived permanent HE cells produced by the methods provided herein are capable of differentiating into hematopoietic lineage cells comprising T cell progenitor cells and T cells in addition to NK cell progenitor cells, NK cells, NKT cells, or B cells.

The invention relates to the field of biology, and discloses a cornu cervi pantotrichum stem cell exosome preparation and a preparation method thereof.The preparation method of the cornu cervi pantotrichum stem cell exosome preparation comprises the following steps that after cornu cervi pantotrichum is disinfected, a semitransparent mesenchymal layer at the tip of the cornu cervi pantotrichum is separated out, the cornu cervi pantotrichum is cut into pieces and then flushed with PBS containing double antibodies, the cornu cervi pantotrichum is put into tissue digestion mixed enzyme, and the cornu cervi pantotrichum exosome preparation is obtained centrifuging and cleaning after digestion, and then spreading in a culture bottle for culture to obtain an isolate; when cells of the isolate grow to 70%-90% of a culture bottle, discarding an original culture solution, cleaning with PBS, adding pancreatin for digestion, terminating digestion with a complete medium, and performing cell passage to obtain a passage; and performing gene editing treatment on the cells of the passage. By obtaining the stem cells of the cornu cervi pantotrichum, damage to a donor when the stem cells are obtained by the donor is avoided, and due to the fact that the cornu cervi pantotrichum stem cells have stable biological characteristics, the difference between different sources is reduced, and gene editing is carried out on the subcultured cells.

Disclosed in the present disclosure is a stable anti-HER2 bispecific antibody, which comprises a first protein functional region and a second protein functional region used for recognizing extracellular domain II and extracellular domain IV of HER2, respectively. The bispecific antibody remains stable after being stored in PBS at about 37°C for 30 days. Further disclosed in the present disclosure are a pharmaceutical composition containing the bispecific antibody and a PD1 antibody, and the use thereof. The bispecific antibody and the pharmaceutical composition containing same of the present disclosure can be used for treating cancers.

The invention discloses a nano compound based on a CRISPR-Cas9 system and application of the nano compound. The nano compound comprises a histidine oligopeptide-cell penetrating peptide conjugate and an RNP compound composed of sgRNA of a target gene and Cas9 protein, and the histidine oligopeptide-cell penetrating peptide conjugate and the RNP compound are self-assembled under the action of zinc ions to form the nano compound. The nano-composite designed by the invention is simple in structure, and overcomes the defects of high cytotoxicity and large particle size (diameter gt; compared with the prior art, the compound has the advantages that the compound can overcome the defects of complex chemical synthesis, potential immunogenicity and the like, can be used for extracellular-cytoplasm-cell nucleus trinity antivirus, is high in virus removal efficiency and good in antivirus effect, and has high market value and development potential.

Disclosed are systems and methods for detecting extracellular ligands and / or inducing expression of an exogenous or endogenous gene. The disclosed systems and methods typically include and / or utilize (i) first and second exogenous extracellular sensors, and third and fourth exogenous extracellular sensors; and (ii) an expression vector comprising a target gene operably linked to a hybrid promoter sequence. The hybrid promoter sequence of the expression vector includes a minimal promoter for inducing transcription and the hybrid promoter sequence further includes interspaced transcription regulator binding sites upstream of the minimal promoter that bind two or more transcription regulators of the extracellular sensors that are released from the extracellular sensors when the extracellular sensor bind an extracellular ligand.

The invention discloses a composition based on an engineered stem cell exosome and application of the composition in treating male sexual dysfunction. The composition comprises a mesenchymal stem cell exosome modified by genetic engineering, and the surface of the mesenchymal stem cell exosome displays a bifunctional fusion polypeptide targeting TbetaR II and LOXL2. According to the fusion polypeptide, TbetaR-PEP and LOXL2-PEP are integrated through a flexible linker, and an HIV-TAT penetrating peptide and a self-assembly motif are introduced, so that nanoparticles are formed, and the half-life period is prolonged. In-vitro experiments show that the inhibition efficiency of the fusion polypeptide on TbetaRII and LOXL2 is obviously superior to that of a single peptide, and the fusion polypeptide can synergistically reverse fibrosis and promote angiogenesis. Animal experiments prove that the exosome loaded with the fusion polypeptide can significantly improve the erectile function of diabetic ED rats. According to the invention, a radical treatment scheme is provided for male sexual dysfunction through double-target cooperation, long-acting delivery and a multi-effect repair mechanism.

The invention discloses a nano antibody and a preparation method and application thereof, the nano antibody comprises a heavy chain variable region, the heavy chain variable region comprises CDR1, CDR2 and CDR3, and the amino acid sequence of the heavy chain variable region of the nano antibody comprises SEQ ID NO: 3. The nano antibody provided by the invention is determined as a competitive OTR inhibitor with extracellular treatment potential; in-vitro and in-vivo experiments prove that the nano antibody has a curative effect equivalent to that of atosiban which is widely used clinically, and a new direction is provided for developing effective anti-premature delivery medicines.

The invention provides a grifola frondosa liquid culture medium as well as a culture method and application thereof, and relates to the technical field of fermentation. The invention provides a grifola frondosa liquid culture medium, which contains glucose, peptone, KH2P2O4, MgSO4, vitamin B1, gibberellin and tyrosol, can significantly improve the growth of grifola frondosa thalli and the secretion of exopolysaccharides, and can be used for the preparation of food, drugs and health care products.

The present disclosure provides an attention-based MIL model for use in extracting and characterising cell features, at both the cell level and the population level. To characterise cells, cells in a well are initially fed into the model. The cells in a well include a points cloud of cells and a point cloud of corresponding cell nuclei. The cells in the well are passed through a pretrained DFN encoder which extrapolates the cell and nuclei features from the cells in the well. These extracted features are then passed through a transformer based encoder to produced transformed versions of the extracted cell and nuclei features. These transformed features are then fed into at least two classifiers. One of the classifiers is a cell-level MLP classifier which is used to compare the cell features with known drug response to attempt to match a phenotype signature with a known drug response. One other classifier is the bag classifier which is used to compare the cell population features with that of known drug responses, again to attempt to match to a phenotype signature. The outputs of the cell classification and the bag classification are used to label the cells in the well with a drug label or a potential drug label / use case.

Disclosed is a single-cell exon sequencing method and a use thereof, the method comprising: S1, tumor single cell sorting; S2, performing MDA amplification of sorted tumor single cells to obtain a genome library; S3, measuring the uniformity of MDA amplification products, selecting multiple sites located on different chromosomes and designing corresponding amplification primers, and testing the amplification effect of the multiple sites of the MDA product of each single cell by means of qPCR, thereby determining whether the genome has been uniformly amplified; S4, gDNA library preparation: breaking the amplified DNA into 150-200 bp fragments, and obtaining a gDNA library by means of end repair, tailing, adding adapters, and amplification; S5, exon capture and library construction: hybridizing the prepared gDNA library with a specific probe, capturing a specific region, and then adding an index tag via PCR amplification to obtain an exon library; and S6, sequencing the exon library.

Disclosed are systems, components of systems, and methods for sensing extracellular ligands and / or modulating expression of an endogenous or exogenous gene in a cell. In some embodiments, the disclosed systems and methods comprise or utilize first and second exogenous extracellular sensors and / or nucleic acid sequences encoding the first and second exogenous extracellular sensors, wherein the first exogenous extracellular sensor comprises: a) a ligand binding domain; b) a transmembrane domain; c) a protease cleavage site; and d) a functional domain comprising an RNA-binding subdomain fused to a transcription regulatory subdomain; and the second exogenous extracellular sensor comprises: e) a ligand binding domain, f) a transmembrane domain, and g) a protease domain that cleaves the protease cleavage site of the first exogenous extracellular receptor.

The invention belongs to the technical field of microorganism application, and particularly relates to a genetically engineered bacterium for producing L-alanine and application of the genetically engineered bacterium in two-stage fermentation of the L-alanine, and the genetically engineered bacterium takes vibrio natrievibrio as a chassis bacterium and can co-express an alanine dehydrogenase gene and an alanine transporter gene. An alanine dehydrogenase gene is heterologously introduced into a vibrio natriticus genome, and an inactivated gene combination with the optimal L-alanine yield is found out by randomly inactivating a key enzyme in a byproduct production pathway, so that the vibrio natriticus genetically engineered bacterium capable of efficiently producing the L-alanine through a fermentation method is constructed. According to the present invention, the L-alanine is produced through the two-stage fermentation by using the gene engineering bacteria through the temperature-sensitive control strategy so as to reduce the influence of the accumulation of the L-alanine on the cell growth, and the high production of the Xiabanmate is provided. The genetically engineered bacterium is short in growth cycle, glucose can be efficiently utilized to convert the genetically engineered bacterium into L-alanine, the L-alanine is transferred to the outside of cells, separation and purification are simple, cell disruption is not needed, and the yield and conversion rate of the L-alanine can both reach a high level.

The invention relates to a modified RNA with a synthetic cap structure and a modified coding region encoding for DNAse1 and / or DNAse1L3. The invention further relates to a saRNA and / or a circular RNA, and to a vector encoding the modified RNA. The invention also relates to a pharmaceutical composition for the use of treating a disease associated with the formation of extracellular DNA, including extracellular traps released from leukocytes, such as NETs.

This invention relates to a method for producing chondroitin with improved safety and extracellular secretion levels, belonging to the field of genetic engineering. Using the probiotic *Escherichia coli* Nissle 1917 as the host, this invention designs methods to increase chondroitin production by studying the chondroitin synthase KfoAC and the chondroitin-linking Kdo synthase. The relationship between chondroitin synthesis and secretion in *E. coli* was clarified, and the specificity of the chondroitin-linking Kdo was investigated, resulting in an extracellular chondroitin production of 0.32 g / L. Subsequently, the introduction of the protein KfoB from the chondroitin synthesis gene cluster increased the extracellular chondroitin production to 0.51 g / L. Further screening and optimization of the expression of transport proteins resulted in an extracellular chondroitin production of 0.73 g / L in shake flasks. Further optimization using a 7-L fermenter resulted in an extracellular chondroitin production of 5.9 g / L.

The invention relates to a circulating tumor cell enrichment, separation and dyeing integrated system and application thereof.The circulating tumor cell enrichment, separation and dyeing integrated system is composed of a bottom plate module, a platform module, a multi-pipette module, a power module and a shell module; the platform module comprises a sample tube rotating and uniformly mixing module, a magnetic adsorption module, a reagent module, a waste liquid pool module and a gun head module, and the sample tube rotating and uniformly mixing module, the magnetic adsorption module and the reagent module are fixedly mounted on a bottom plate I in sequence. The CTC separation and dyeing integrated system is developed on the basis of the immunomagnetic bead technology, enrichment separation and immunofluorescent dyeing of CTC can be automatically completed by running a program, manual operation errors are effectively avoided through a programmed process, and the CTC separation and dyeing integrated system is particularly suitable for clinical CTC detection. The system is also suitable for other enrichment and separation applications based on an immunomagnetic bead technology, including but not limited to enrichment and separation of biological macromolecules such as cells, exosomes, proteins and the like.

The invention relates to the technical field of plant genetic engineering, and particularly provides cloning of a GbGELP25D gene of gossypium barbadense and application of the GbGELP25D gene of gossypium barbadense. According to the invention, the GbGELP25D gene with a full length of 1092 bp is cloned from the sea island cotton variety Xinhai No. 7 for the first time, and the gene encodes a secretory lipase protein with a signal peptide and is positioned in an extracellular gap. Through bioinformatics analysis, phylogenetic classification, protein structure modeling and signal peptide function verification, it is clear that the gene belongs to a plant GELP family. Furthermore, a virus-induced gene silencing technology is utilized to prove that the silent GbGELP25D can obviously enhance the resistance of cotton to verticillium wilt, and the mechanism of the silent GbGELP25D is closely related to activation of ethylene synthesis and signal channels and induction of expression of disease-resistant related genes. The resistance gene provided by the invention enriches gene resources of cotton verticillium wilt resistance breeding, and has important theoretical significance and application value.

To provide a new method, model, program or the like for acquiring information usable for a disease diagnosis or prognosis prediction by using a sample taken from lungs or exhaled breath.SOLUTION: A method for assisting disease diagnosis or prognosis prediction including: acquiring (A) extracellular particle information on a population of extracellular particles collected from lungs or exhaled breath of a subject; the extracellular particle information including scattered light information on scattered light of irradiated light about each extracellular particle of the population of extracellular particles, and light emission information on a plurality of light emission whose peak wavelengths are mutually different, caused from a component of the extracellular particles or a labelling substance connected to the component due to the irradiated light; and (B) disease information on a state of a disease in the subject from each subject included in a group of subjects; and acquiring information for assisting the disease diagnosis or the prognosis prediction related to the extracellular particles collected from the lungs or the exhaled breath by machine learning on the basis of a data set including the extracellular particle information and the disease information.SELECTED DRAWING: Figure 1

The present invention provides a method for culturing organoids, the method comprising: a) disassociating unprocessed organoids to produce a cell suspension; b) sieving the cell suspension through a cell strainer to retain a sieved cell suspension containing cells of about 10 μm to about 1 mm in diameter; and c) seeding cells of the sieved cell suspension into a bioreactor in a cell culture medium comprising an extracellular support matrix.

The invention provides a nano drug delivery system, which comprises (a) an active component, which comprises an anti-tumor active substance or a mixture of the anti-tumor active substance and a substance capable of synergistically generating an anti-tumor effect with the anti-tumor active substance; (b) a carrier material, wherein the carrier material comprises an mPEG-PLGA-HA material which is modified or not modified with a PD-1 or PD-L1 monoclonal antibody; optionally (c) a lipid material selected from the group consisting of lecithin (PC), cephalin (PE), mPEG-PE, mPEG-DSPE, mPEG-DPPE, DOP-DEDA, or a combination thereof; and the active ingredient is wrapped by the carrier material. The nano drug delivery system has transmembrane capability, and can deliver active components to the outside of cells, cytoplasm and / or cell nucleus.

The invention relates to the technical field of biological medicines, in particular to a preparation method of protein cross-linked nano affinity microspheres, which comprises the following steps: preparing carboxylated silicon dioxide nano microspheres with the particle size of 500nm by adopting a sol-gel method, mixing tetraethoxysilane and ammonia water according to the volume ratio of 1: (3 + / -1), reacting in an ethanol solution at 40 DEG C for 6 hours, filtering, washing, and drying to obtain the protein cross-linked nano affinity microspheres. And centrifuging to obtain a microsphere precipitate, washing with deionized water for three times to finally obtain a nano microsphere suspension of which the surface is rich in carboxyl, and controlling the solid content to be 10 + / -0.5 mg / mL. The capturing efficiency of the finally prepared protein cross-linked nano affinity microspheres for exosomes reaches 95.0%, the binding capacity reaches 28.5 mu g / mg, the purity of the extracted exosomes is higher than that of an ultracentrifugation method, the biological activity and structural integrity of animal cell exosomes and plant cell exosomes are completely kept, and the protein cross-linked nano affinity microspheres are suitable for large-scale industrial production. The problems that in a traditional exosome extraction method, the recovery rate is low, impurity pollution is serious, exosome activity is damaged, and the application range is limited are solved.

The invention discloses a method for quantifying response of two carbon utilization modes of microalgae to weathered carbon sink of magnesium-containing silicate rock, which comprises the following steps: culturing two groups of microalgae under the same culture condition, adding magnesium-containing silicate rock and microalgae into one group, and setting an inhibitor (AZ / DIDS) added with a certain carbon utilization mode as an experimental group; and only magnesium-containing silicate rock and microalgae are added into one group as a blank group without adding an inhibitor (AZ / DIDS) of a certain carbon utilization mode. Setting the treatment group added with AZ (10mM) and DIDS (2mM) as an experimental group, and subtracting the weathering quantity of the magnesium-containing silicate rocks in the treatment group not added with AZ / DIDS and the treatment group added with AZ / DIDS to obtain the weathering quantity delta Ti of the magnesium-containing silicate rocks under the influence of the microalgae anion channel utilization mode, namely delta Ti = Ti-Ti (AZ / DIDS), and rock weathering carbon sink Qi = delta Ti under the influence of the microalgae anion channel utilization mode. Fitting a polynomial function Q = Q (i) to obtain the response function of the microalgae extracellular carbonic anhydrase mode or the microalgae anion channel utilization mode to the weathering carbon sink of the magnesium-containing silicate rock. According to the method, the rock weathering carbon sink quantity under the influence of two carbon utilization modes of the microalgae can be rapidly and accurately obtained. The invention provides the method for quantifying the response of the two carbon utilization modes of the microalgae to the weathered carbon sink of the magnesium-containing silicate rock so as to overcome the defect that the response of the two carbon utilization modes of the microalgae to the weathered carbon sink of the magnesium-containing silicate rock cannot be quantified in the prior art.

The present invention provides molecules that bind to cellular components or parts thereof (e.g., filaments or histones forming the cytoskeleton or nucleoskeleton) that are exposed to the extracellular environment upon cell death, which are specifically observed in diseased tissues, and simultaneously bind to target molecules on or in cells that act specifically in tissues where cell death is observed, such as diseased tissues, abnormal tissues, etc. The molecules of the present invention are useful as drugs that provide significant therapeutic or preventive effects while reducing side effects.