398 results about "Cytoplasm" patented technology

The present disclosure provides bispecific chimeric antigen receptors that target BCMA and CD19. The CAR may comprise an scFv targeting BCMA and an scFv targeting CD19, a hinge region, a transmembrane domain, a co-stimulatory region, and a cytoplasm signaling domain. Chimeric antigen receptors can be used to treat autoimmune disorders or cancer.

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 multimodal-based cell experiment image processing method and system and a storage medium, and the method comprises the steps: obtaining a single-channel gray image or a multi-channel fluorescence image containing cytoplasm and cell nucleus characteristics, carrying out the Gaussian filtering of a cytoplasm channel, and carrying out the median filtering of a cell nucleus channel; performing threshold segmentation on the preprocessed cytoplasm channel image by using a maximum between-class variance method to generate an initial cytoplasm mask; performing adaptive threshold segmentation on the preprocessed cell nucleus channel image to generate an initial cell nucleus mask; performing logic OR operation on the initial cytoplasm mask and the initial cell nucleus mask to generate an initial cell region; and finally, calculating each morphological parameter of each cell region and removing the cell regions of which the morphological parameters exceed a preset range to generate a segmentation result of the complete cell region, the morphological parameters and spatial distribution information. The problems that in the prior art, cytoplasm and cell nucleus noise processing segmentation precision is low, overlapped nucleus regions are difficult to separate, and morphological parameter analysis reliability is poor are solved.

The invention discloses a stem cell fusion degree detection method and system based on artificial intelligence, and a storage medium. The method comprises the following steps: carrying out image preprocessing on a cell microscope image; automatically calculating an optimal threshold value by using an image threshold value segmentation algorithm to obtain a cytoplasm mask; median filtering is carried out on the original image to reduce noise, then an adaptive threshold segmentation method is adopted, a local threshold is calculated according to local area gray level distribution, and a cell nucleus binary image is generated; performing connected region marking on the cell nucleus binary image, calculating the area attribute of each region, and performing filtering according to a cell nucleus removal ratio parameter to obtain a cell nucleus mask; performing logic OR operation on the cytoplasm mask and the cell nucleus mask to obtain a complete cell segmentation result; and calculating the fusion degree of the stem cells based on the cell segmentation result. Therefore, the problems of accuracy and consistency of judging the fusion degree of the stem cells by observing microscope images with human eyes in the prior art are solved, and accurate detection of the fusion degree of the stem cells is realized.

The invention relates to the field of image processors and biological medicines, and discloses an NK cell activity rapid detection method based on image processing. The method comprises the following steps: acquiring an unmarked time sequence phase image sequence of an NK cell and target cell co-culture system; performing cell instance segmentation to track individual cells; extracting a morphological dynamic characteristic parameter set of the target cell, wherein the morphological dynamic characteristic parameter set comprises a volume change rate, a phase gradient entropy, a cytoplasm phase fluctuation frequency and a nuclear region phase mean value; inputting the parameters into a pre-trained death state discrimination model, and outputting a death probability; and calculating a killing efficiency index based on the death probability evolution curve, and judging the activity level of the NK cells. The system comprises a phase image acquisition unit, a cell segmentation unit, a feature extraction unit, a death judgment unit and an activity judgment unit. Through unmarked imaging and deep learning fusion analysis, high-precision, real-time, quantitative and ultra-early NK cell activity evaluation is realized, and the method is suitable for clinical instant inspection and immunotherapy monitoring.

The invention provides a cell protein degradation platform based on an artificial biological macromolecular aggregate. Specifically, the invention provides a PROTAC functional module based on an interworking nucleic acid skeleton, a PROTAC-aggregate complex (MLO-PROTAC), a kit, application and a preparation method of the PROTAC functional module, the PROTAC-aggregate complex (MLO-PROTAC) and the kit, and also provides a targeted protein degradation method. In a PROTAC platform built by the PROTAC functional module and the PROTAC-aggregate complex, a programmable nucleic acid component is used as a core assembly unit, and the functional module is enriched and spatiotemporal-spatial regulation is performed by using the polypeptide aggregate, so that the delivery efficiency and the cytoplasm exposure degree are remarkably improved while the universality is maintained, and the delivery efficiency and the cytoplasm exposure degree are remarkably improved. Therefore, a more effective target protein degradation way is provided for the field.

The invention discloses a liquid cell product, a preparation method thereof and a preserving fluid for preparing the liquid cell product, and belongs to the technical field of immunodetection. The dendritic polymer is creatively added into the preserving fluid for preparing the liquid cell product, the cell morphology in the liquid cell product can be improved, and a thermal acceleration test proves that the preserving fluid can maintain the cell morphology in the liquid cell product for a long time. When the preservation solution is used for preparing the liquid cell quality control product for immunodetection, quality control spots with complete circles can be formed after sample application on slides made of different materials, it can be obviously judged that cells in the liquid cell quality control product are dispersed very uniformly after immunohistochemical staining, and the preservation solution has very high application value.

The invention discloses a drug delivery system based on branched chain nucleic acid as well as a preparation method and application of the drug delivery system. The drug delivery system comprises a multivalent nucleic acid framework, a nucleic acid-small molecule coupling drug and a nucleic acid-polypeptide coupling drug, a nucleic acid nanostructure is formed through base complementary pairing assembly, and the nucleic acid-small molecule coupling drug comprises first nucleic acid and an E3 ubiquitin ligase ligand coupled with the first nucleic acid; the nucleic acid-polypeptide coupling drug comprises a second nucleic acid and a cell penetrating peptide coupled with the second nucleic acid. The drug delivery system disclosed by the invention can respond to a tumor microenvironment, so that cell penetrating peptides are activated, tumor penetrating and cytoplasm delivery capacities are enhanced, the aim of efficiently degrading tumor-related target proteins is fulfilled by combining a multivalent effect, and a remarkable tumor treatment effect is achieved; in addition, the preparation method is simple and easy to implement, has universality and high production efficiency, and is expected to be applied to treatment of various tumors.

The invention discloses a Yarrowia lipolytica engineering bacterium for producing astaxanthin in a peroxisome compartment and application of the Yarrowia lipolytica engineering bacterium. A complete astaxanthin synthesis path is expressed in host bacteria by recombinant Yarrowia lipolytica; comprising geranyl geranyl diphosphate synthase CrtE, phytoene synthetase / lycopene cyclase CrtYB, phytoene desaturase CrtI, 3-hydroxy-3-methylglutaryl CoA reductase tHMGR, beta-carotene ketolase CrtW and beta-carotene hydroxylase CrtZ, and the enzymes are relocated into peroxisome, so that the enzyme activity of the peroxisome is improved, and the enzyme activity of the peroxisome is improved. The number of expression hemoglobin genes VHb and peroxisome is increased, the surface area of the hemoglobin genes VHb is enlarged, and one or more of enzymes PEX11, PEX19, DNM1 or PHO85 for prolonging the service life of the hemoglobin genes VHb and peroxisome is / are obtained; the astaxanthin production performance of the recombinant strain is verified based on cytoplasm engineering and subcellular organelle compartment engineering, and the astaxanthin production capacity of the yarrowia lipolytica is further improved.

The invention discloses a cherry sugar transporter gene CpSWEET10 and application thereof, and belongs to the technical field of cherry gene engineering. A novel sugar transporter gene CpSWEET10 is obtained through cloning, the nucleotide sequence of the novel sugar transporter gene CpSWEET10 is shown as SEQ ID NO.1, and the coded amino acid sequence of the novel sugar transporter gene CpSWEET10 is shown as SEQ ID NO.2. The gene is highly expressed in cherry fruits, and the content of soluble solids and total sugar in mature fruits is remarkably positively correlated with the expression quantity of the soluble solids and the total sugar; through subcellular localization and sugar transport function defect type yeast mutant functional complementation experimental analysis, the protein coded by the CpSWEET10 gene is localized in a cytoplasmic membrane and has a sucrose transport function; the cherry sugar transporter gene CpSWEET10 disclosed by the invention can be applied to the improvement of the sugar degree character of the fruits, and a new gene resource is provided for molecular breeding for improving the high-sugar quality of the cherry fruits.

The invention relates to the technical field of cell in-vitro culture. The invention aims to provide an amplification method of high-purity anti-tumor NK cells. The amplification method comprises the following steps: S1, obtaining initial NK cells; s2, performing double-stage culture; a double-stage culture mode is adopted, the first-stage culture period is 8 days, the second-stage culture period is 10 days, and after the first-stage culture is finished, NK cells with full cytoplasm are separated through a density gradient centrifugation method for second-stage culture; s3, harvesting the NK cells. The method has the advantages of simplicity and convenience in operation, controllable cost, suitability for large-scale production and the like, and a more efficient and economical solution can be provided for preparation of the high-purity anti-tumor NK cells.

A method of pretreatment of a sample containing a cell for mass spectrometry, including contacting the cell with a first acidic solution containing an organic acid; and extracting a cytoplasmic component of the cell by heating the cell in contact with the first acidic solution.

The invention discloses a mechanical response type engineered mesenchymal stem cell exosome preparation for treating diseases caused by fibrosis inflammation, a preparation method and application, and the exosome preparation is an exosome which is secreted by mesenchymal stem cells and has high cholesterol and high-efficacy protein. The exosome is prepared by culturing mesenchymal stem cells on a hydrogel matrix in a modulus hardness range of 1-50KPa, the exosome secretion amount of a single cell is (3-5) * 10 < 4 > / cell, the cholesterol content is (1-10) nM / 5mg exosome, and the size of the exosome is 110-140nm; the high-efficacy protein is a key functional protein for regulating and controlling the biological activity of the exosome and the cell action efficiency. The cholesterol content in the exosome is precisely regulated and controlled, the cytoplasm delivery efficiency and tissue penetrability of the exosome are improved, a whole-process control system of hydrogel preparation, stem cell culture and exosome purification is constructed, and the stability of the preparation and the yield of the exosome are improved.

The invention relates to the field of medical image processing, and particularly discloses a TLS structure sketching system and method based on artificial intelligence, and the method comprises the steps: S1, image preprocessing: carrying out the standardization processing of an input HE staining section image, firstly separating cell nucleus and cytoplasm staining components through a color deconvolution algorithm, and highlighting the nucleoplasm contrast of lymphocytes; then strengthening the cell contour boundary by adopting an edge detection algorithm, and connecting the fracture edge through morphological operation to form a continuous and clear cell boundary mask; s2, morphological feature extraction: performing single cell segmentation based on the preprocessed image, and extracting geometric features and texture features of each cell; through machine learning model training, distinguishing lymphocytes and non-lymphocytes according to the characteristics, and generating a lymphocyte distribution probability graph; and S3, preliminarily identifying the aggregated area. By adopting the technical scheme of the invention, the TLSs differentiation stage can be identified, and the identification accuracy can be improved by combining morphological characteristics and spatial distribution characteristics.

Systems and methods for multi-modal subcellular segmentation using photolysable biomarkers and / or transcriptomic readout density maps are disclosed. The systems and methods improve the accuracy of cell segmentation of the nucleus, cytoplasm, and cell membrane regions by using optical and bleach correction from a variety of photolysable morphological markers in combination with high quality 3D images acquired with high dynamic range scans and spatial transcriptomic readout density maps.

The invention discloses an amniotic epithelial stem cell screening method and device based on microscopic image feature extraction, relates to the technical field of microscopic image analysis and cell screening, and is used for solving the problem that group topology and internal activity in amniotic epithelial stem cell lossless screening are difficult to collaboratively quantify. The method comprises the following steps: firstly, performing enhancement processing on a phase difference microscopic image, and extracting a kernel centroid and a global gradient energy diagram; then, constructing a Voronoi jurisdiction, and coupling a cytoplasm texture entropy and a gray scale attenuation gradient to generate a cytoplasm compactness factor; meanwhile, calculating a cell gap topology consistency index based on mapping of the Voronoi boundary and the gradient map; and finally, weighting and correcting the compactness factor by using a topological index to obtain a dryness maintenance confidence coefficient, and outputting a target coordinate. Through dual verification of geometric topology and physical gradient, combined constraint screening of internal and external characteristics of cells is realized.

The invention discloses application of fluorinated ionizable lipid as a gene drug delivery carrier, and belongs to the technical field of biological medicine. The gene drug delivery carrier provided by the invention is prepared from fluorinated ionizable lipids, cationic lipids, auxiliary lipids, structural lipids and PEG lipids, wherein the molar percentage of the fluorinated ionizable lipids in the gene drug delivery carrier is 1%-10%. According to the invention, the molar percentage of the fluorinated ionizable lipid in the total lipid is optimized to 1-10%, so that the cytotoxicity is obviously reduced, and more excellent nucleic acid transfection efficiency is realized in vivo and in vitro; various nucleic acid drugs including DNA, mRNA, siRNA and CRISPR-Cas9 ribonucleoprotein can be efficiently delivered, action targets of the nucleic acid drugs are expanded to cell nucleuses and cytoplasm, and the nucleic acid drugs are more accordant with the action mechanism of modern gene therapy and editing.

The invention belongs to the technical field of biomedical materials, and particularly relates to nucleic acid for synthesizing human III-type collagen as well as an in-vivo expression method and application of the nucleic acid. The nucleic acid sequence of the synthesized human III type collagen is as shown in SEQ ID NO. 1. The invention provides an effective mRNA (messenger ribonucleic acid) nucleic acid, the nucleic acid encodes a human full-length III-type collagen gene sequence, can effectively realize expression in a human body, and is guided to translate into full-length III-type collagen through the nucleic acid in human cytoplasm and has a correctly folded space structure triple-helix collagen, namely human III-type collagen.

The application discloses a mucous membrane adhesion auxiliary material and a lipid material with mucous membrane adhesion prepared by using the auxiliary material, the nanoparticle has mucous membrane adhesion and can be adhered to the surface of a mucous membrane and is suitable for mucous membrane administration. The lipid nanoparticle promotes the entry of the lipid nanoparticle into cells by forming a disulfide bond with the sulfydryl on the surface of mucous membrane cells, and the entry of the lipid nanoparticle into cells is a non-endosome-lysosome pathway, so that the encapsulated nucleic acid macromolecule can be avoided from being degraded by enzymes in cells, thereby directly delivering the macromolecular nucleic acid in the cytoplasm to play a role.

This invention discloses a biodegradable brush-shaped polymer drug carrier and its applications, belonging to the field of polymer compound technology. The brush-shaped polymer drug carrier is obtained by a click reaction between a polydisulfide backbone and cationic side chains. The polydisulfide backbone in its structure can undergo a special "sulfhydryl exchange" reaction with free thiol groups present on cell membrane proteins, directly carrying the nucleic acid drug into the cell through the cell membrane, thus completing the delivery of the nucleic acid drug. After entering the body, under the action of glutathione (GSH) which is abundant in the tumor cell cytoplasm, the polydisulfide backbone is broken and degraded, releasing the nucleic acid drug, which is suitable for the delivery of nucleic acid drugs.

The application discloses a single-cell biopsy and dynamic transcriptome tracking system and method, and belongs to the technical field of single-cell sequencing. The system comprises the following steps: culturing single cells to be detected in a cell culture dish; puncturing the single cells by using an amino-modified quartz nanocapillary to extract trace cytoplasm samples; recovering the single cells in a culture environment; applying specific stimulation to the recovered single cells; at one or more time points after the stimulation is applied, puncturing the same single cells again by using the quartz nanocapillary to extract trace cytoplasm samples, and recovering the single cells after each extraction; and respectively performing reverse transcription, amplification, library construction and sequencing on the cytoplasm samples extracted at different time points to obtain dynamic transcriptome data of the single cells at different time points. The application can perform low-damage multiple longitudinal biopsies on the same single living cell, and can construct a dynamic transcriptome atlas of the single cell on a time axis in combination with downstream sequencing.

The application discloses a kind of artificial cells for single-cell mass spectrometry and preparation method thereof, wherein artificial cell includes internal water phase, intermediate oil phase and external water phase;Internal water phase includes polyethylene glycol and polyvinyl alcohol aqueous solution, intermediate oil phase includes L-alpha-phosphatidylcholine chloroform and hexane mixture;External water phase includes PVA and F-68 aqueous solution.Compared with natural cell sample, the novel artificial single cell based on microfluidic self-assembly developed in the application has better uniformity, stability and controllability, effectively avoids significant measurement difference between single-cell individual samples, and effectively solves the problem of difficult stable preservation of biological samples.The proposed artificial cell preparation method will largely solve the problem of lack of standard reference material in the field of single-cell mass spectrometry, making the results of single-cell mass spectrometry method more accurate, reliable and mutually recognized.

Myotonic Dystrophy type 1 (DM1) is an inherited disease characterized by multi-systemic symptoms, particularly in skeletal muscles (progressive weakness and atrophy, myotonia). The inventors screened 7 500 bioactive compounds for their ability to improve the myogenic fusion and reduce the molecular hallmarks of DM1 skeletal muscle cells. The inventors found that five compounds, all inhibiting HDAC6, a cytoplasmic histone deacetylase, were effective at the micromolar range in normalizing the myogenic fusion, reducing the number of nuclear foci of mutant DMPK mRNA, decreasing the expression level of DMPK mRNA, restoring the splicing of several genes, and increasing the acetylation of SMAD3, a transcription factor involved in muscle development. The effects of HDAC6 inhibitors were observed both in immortalized and hiPSC-derived skeletal muscle cells from DM1 patients, and in different stages of differentiation. Thus, the present invention relates to the use of selective HDAC6 inhibitors for the treatment of DM1.

The present application relates to a kind of drug composition containing STING agonist and WIP1 inhibitor and its liposome and application.The composition can be co-encapsulated in liposome with STING agonist and WIP1 inhibitor, form stable drug delivery system.The liposome can promote efficient endocytosis of cell, and release two active ingredients in cytoplasm synchronously, block the negative feedback mechanism of STING signal path by WIP1 inhibitor, enhance and prolong the activation level of STING path, to synergistically inhibit tumor growth.Based on the mechanism, the drug composition of the present application can be used for preparing the drug for treating diseases related to STING path (such as tumor).

The invention discloses a blocking agent for cell immunofluorescence detection by a slide method and a use method of the blocking agent. The blocking agent is a homologous no-load cell lysis solution without transfection plasmids, and the protein concentration is greater than or equal to 30mg / mL. The blocking agent is completely the same as cell cytoplasm and cell membrane components of cells to be detected except for plasmids-expressed antigens, when in use, the blocking agent is pre-mixed with a blood cleaning sample in advance, interfering antibodies except target antibodies in serum can be purified, and meanwhile, due to the fact that no antigen protein expressed after plasmid transfection exists, the blocking agent can be used for detecting the cells to be detected. And the protein concentration is equivalent to the globulin concentration in serum, so that the target antibody cannot be lost in the serum, and the slide method CBA detection sensitivity can be greatly improved.

The invention discloses a histone-derived peptide nano-particle for targeted degradation of cytoplasm cGAS and a preparation method and application thereof, and is characterized in that a polypeptide compound is dissolved in a phosphate buffer, ultrasonic treatment and room temperature standing self-assembly are performed to form nano-particles, and centrifugal purification is performed to obtain the histone-derived peptide nano-particle A4-CMA NPs for targeted degradation of cytoplasm cGAS. The nano-particles (A4 at CMA NPs) can be combined with cytoplasm cGAS in a targeted manner and mediate the cytoplasm cGAS to degrade, so that the activation of a cGAS-STING pathway is blocked, periodontal tissue inflammation and alveolar bone resorption are effectively relieved, and a new strategy is provided for periodontitis treatment.

The invention discloses an Apopsin3 mutant and a method for regulating an environment in a plant cell to be acidic, and belongs to the technical field of pH regulation in the plant cell. The regulation and control method comprises the following steps: mutating 83rd and 120th amino acids of the light-operated hydrogen ion pump protein Apopsin3, converting into hydrogen ion channels, expressing the hydrogen ion channels on a cell membrane in a targeting manner, and stimulating with green light of 40-500 [mu] W / mm < 2 > and 520-560 nm so as to regulate the pH in the cell to be acidic. By adopting the cell specific promoter and performing site-specific mutagenesis modification on the specific hydrogen ion pump, accurate pH regulation and control on specific cells or organelles can be realized. An Apopsin3 (FARH) gene is fused with a cell membrane targeting sequence, and under the stimulation of specific light, only the pH of cytoplasm is obviously changed, and the pH of other cell regions is not influenced.

The application discloses application of branched-chain amino acid transaminase 1 gene in treatment of maple syrup urine disease, and relates to the field of gene therapy. Maple syrup urine disease caused by BCAT2 gene defect can be effectively treated by up-regulating branched-chain amino acid transaminase 1 (BCAT1), and it is verified for the first time that overexpression of BCAT1 (cytoplasm type / brain type) can systemically save lethal metabolic phenotypes caused by BCAT2 (mitochondrial type / general type) defects. The application breaks the barrier of metabolic division, and proves that expression of non-mainstream metabolic enzyme (BCAT1) in non-classical metabolic organs (liver) is enough to reconstruct BCAA decomposition flux. The application expands the MSUD treatment target spectrum: from “must repair BCKDC” to “can compensate for upstream limiting steps in ectopia”, and provides alternative strategy reserves for BCKD-deficient MSUD.

The invention relates to a formulation for total and differential counting of leukocytes for use in clinical analyses, in particular hemograms, to promote instantaneous differential staining of leukocytes in liquid medium, allowing greater practicality and agility in the total and differential counting of leukocytes. The practicality and agility are related to the use of a single dye for two parameters, eliminating the need for a smear. It results in a satisfactory effect for staining cells in suspension, staining the nucleus and cytoplasm of leukocytes in different shades, allowing their visualization under an optical microscope or in Point-of-Care image processing devices. It uses easily accessible raw materials, including cresyl acetate violet, ethyl alcohol, sodium and potassium chlorides, sodium hydroxide, acetic acid, triton X and deionized water, presenting a simple manufacturing process and low cost.

The present disclosure provides an ionizable lipid that can be used for nucleic acid delivery into cytoplasm.An ionizable lipid of the present disclosure is, for example, a compound represented by the following formula (I) or a pharmaceutically acceptable salt thereof.whereinL1 represents —(CH2)n—,L2 represents —(CH2)m—,n represents an integer of 1 to 5,m represents an integer of 1 to 5,X1 and X2 each independently represent —OC(O)— or —OC(O)O—,Y represents —OC(O)—, or —OC(O)O—,R1 and R2 each independently represent an alkyl group having 2 to 25 carbon atoms or alkenyl group having 2 to 25 carbon atoms that is optionally substituted with one or more of alkoxy groups having 4 to 12 carbon atoms, R1 or R2 each have a total of 4 to 30 carbon atoms, andP is represented by the following formula P-1 or formula P-2, andR3 represents an alkyl group having 1 to 5 carbon atoms that is optionally substituted with a hydroxy group,R4 and R5 each independently represent a hydrogen atom or an alkyl group having 1 to 5 carbon atoms, or R4 and R5 are taken together to form an alkylene group having 2 to 5 carbon atoms,p represents 0 or 1, q represents an integer of 0 to 2, R6 and R7 each independently represent an alkyl group having 1 to 5 carbon atoms,r represents an integer of 1 to 5, and* represents a linking site.