461 results about "Extracellular" patented technology

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 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.

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.

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.

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 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 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 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 invention provides a spot universal CFR64-T cell prepared on the basis of CRISPR / Cas9 as well as a preparation method and application thereof. The CFR64-T cell is obtained by induced differentiation of recombinant human embryonic stem cells; in the recombinant human embryonic stem cell, a coding sequence of a CFR64 molecule is knocked into a TRAC site of the human embryonic stem cell at a fixed point through a CRISPR / Cas9 editing system; the CFR64 molecule comprises an antigen binding structural domain, a transmembrane structural domain and an intracellular costimulatory signal structural domain, the antigen binding domain is a human Fc gamma receptor extracellular domain, and the amino acid sequence of the antigen binding domain is shown in SEQ ID NO: 1. The CRISPR / Cas9 technology is adopted, gene integration sites are controllable and free of oncogene mutation risks, TCR genes are knocked out, the immunological rejection risk of universal CFR64-T cells to hosts is reduced, and meanwhile gene expression is more stable.

The application discloses a kind of multi-effect repair type collagen and its application in the technical field of synthetic biology, the application selects transdermal peptide TD-1, the coding gene of the transmembrane region of human XVII type collagen, extracellular sixteenth non-collagen region and extracellular fifteenth collagen region and His tag is connected in series, and gene sequence is optimized by pichia pastoris codon selection preference, then obtains T-COL17R3 by construction and expression. Through efficacy experiment, it is verified that T-COL17R3 compared with similar products on sale, not only excellent transdermal performance, also has more optimal ability of promoting cell proliferation, migration, anti early glycation product ketone amine and anti late glycation product dicarbonyl compound, better free radical scavenging ability and anti-elastase ability, and can reduce the synthesis of melanin in B16-F10 cell, therefore, T-COL17R3 has excellent application potential in the development of drug composition or skin care product with repair, anti-wrinkle firming or whitening effect.

The invention provides a method for producing L-carnosine, a genetic engineering strain as well as a preparation method and application of the genetic engineering strain, and belongs to the field of genetic engineering. According to the genetic engineering strain provided by the invention, by knocking out a glucose-6-phosphate isomerase coding gene pgi, knocking out an L-threonine / L-homoserine transporter coding gene rhtA or knocking out a dipeptide transfer protein coding gene dppABCD, the yield of extracellular L-carnosine is increased, the shake flask yield of the finally constructed strain reaches 9.1 g / L, the 5L fermentation tank yield reaches 65 g / L, and the yield of the extracellular L-carnosine reaches 9.1 g / L; and the method has a good application prospect in L-carnosine biosynthesis.

A physical method for regulating molecular transport in the extracellular space of the brain includes applying an external pressure to the brain tissue of an animal, and the rhythm of applying the external pressure is associated with the automaticity of the animal. A device for regulating molecular transport in the extracellular space of the brain includes a detection mechanism, a pressure mechanism, and a control mechanism. The detection mechanism is capable of detecting the automaticity of the animal. The pressure mechanism is capable of applying an external pressure to the brain tissue of the animal. The control mechanism is capable of controlling the pressure mechanism based on the detection result of the detection mechanism such that the rhythm of applying the external pressure of the pressure mechanism is associated with the automaticity of the animal. The method and device can effectively regulate molecular transport in the extracellular space of the brain.

The invention discloses a differential low-noise integrated biosensor array for cell culture detection and a preparation method thereof, and the differential low-noise integrated biosensor array integrates an ISHFET device, an REHFET device and an EXHFET device, so that extracellular action potential signals and local field potential signals can be detected during in-vitro cell culture; therefore, interference of factors such as tissue trauma and immune response on cell activity signal detection is avoided, and good reliability is realized; the ISHFET device and the REHFET device can form a differential sensing unit, so that the effect of reducing the detection error is realized; the EXHFET device not only can detect extracellular action potential signals, but also can independently detect local field potential signals, so that more diversified cell activity signals can be detected, and more data support is provided for in-vitro cell culture research. The method is widely applied to the technical field of semiconductors.

This invention provides a chimeric antigen receptor (CAR) targeting both BCMA and GPRC5D antigens and its uses. The chimeric antigen receptor (CAR) includes an extracellular localization signaling domain, an antigen domain targeting BCMA, an antigen domain targeting GPRC5D, a hinge region, a transmembrane region, a co-stimulatory factor, and an intracellular CD3ξ signaling domain. The antigen domain targeting BCMA includes a heavy chain variable region with an amino acid sequence as shown in SEQ ID NO: 1 and a light chain variable region with an amino acid sequence as shown in SEQ ID NO: 2. The antigen domain targeting GPRC5D includes a light chain variable region with an amino acid sequence as shown in SEQ ID NO: 3 and a heavy chain variable region with an amino acid sequence as shown in SEQ ID NO: 4. The dual chimeric antigen receptor, including an antigen domain targeting BCMA and an antigen domain targeting GPRC5D, can simultaneously recognize two anti-tumor targets, preventing tumor immune escape.

The invention discloses bacillus subtilis for recombinant expression of a hermetia illucens HIAMP0438 gene and a construction method of the bacillus subtilis, and belongs to the technical field of gene engineering. The hermetia illucens recombinant protein used in the invention is obtained by expression of bacillus subtilis WB800N transferred into a pHT43-HIAMP0438 high-efficiency expression vector, and the pHT43-HIAMP0438 high-efficiency expression vector is obtained by a homologous recombination technology, so that high-level expression of the HIAMP0438 gene is realized, the HIAMP0438 gene can be efficiently secreted out of cells, the preparation process of the recombinant protein is greatly simplified, and the production cost is reduced. According to the present invention, the recombinant protein can significantly inhibit the growth of Escherichia coli, Salmonella and Listeria monocytogenes, and the recombinant protein can be used as the animal health product, and can be used for preventing and treating pig and chicken intestinal diarrhea caused by Escherichia coli and Salmonella and food-borne digestive tract diseases caused by Listeria monocytogenes.

The application discloses a low-oxygen and pH dual-response nano-carrier, a drug-loaded nanoparticle and application thereof, and belongs to the technical field of biological medicines. The nano-carrier with moderate particle size and low-oxygen and pH dual-response is obtained by using the reaction of aldehyde benzoic acid, nonaethylene glycol and azo-p-phenetidine, the nano-carrier is specifically targeted to a tumor site, and then specifically releases drugs, thereby reducing toxic side effects; meanwhile, after the nano-carrier responds, benzaldehyde groups are released, the benzaldehyde groups are covalently connected with proteins on a lysosome membrane, the structure and activity of the proteins are changed, the lysosome membrane is broken after the proteins are denatured, the permeability of the lysosome membrane is increased, the nano-carrier cannot be discharged to the extracellular, the drug concentration in cells is greatly improved, and the drug bioavailability of the nano-carrier is significantly improved; in addition, the drug loading capacity of the nano-carrier is high, and therefore, the nano-carrier has a good application prospect.