207 results about "Cytoplasm" patented technology

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 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.

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 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.

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 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 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.

The application discloses application of pentagalloyl glucose (PGG) in preparation of an anti-doxorubicin cardiotoxicity medicine. The process is as follows: a myocardial cell H9C2 injury model is induced and established by doxorubicin (DOX), and a minimum concentration of the myocardial cell injury induced by DOX is determined. After 20 muM of PGG is added to a treatment group, DOX-induced H9C2 cell apoptosis can be significantly improved. A mouse myocardial injury model is induced and established by DOX, compared with a doxorubicin group, the left ventricular systolic fraction (FS%) and the ejection fraction (EF%) of mice in the doxorubicin+pentagalloyl glucose group are significantly increased, the myocardial fiber structure is improved, cytoplasm vacuolization is reduced, the serum cTnT and BNP levels are reduced, and the ROS level is reduced. It is proved by the above experiments that the pentagalloyl glucose can significantly reduce DOX-induced myocardial toxicity.

The application discloses an amnion 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 the population topology and internal activity are difficult to be quantified in the lossless screening of amnion epithelial stem cells. Firstly, phase contrast microscopic images are subjected to enhancement processing, and nucleolus centers and global gradient energy maps are extracted; subsequently, Voronoi districts are constructed, and cytoplasm texture entropy and gray attenuation gradient are coupled to generate a cytoplasm compactness factor; meanwhile, a cell gap topology consistency index is calculated based on the mapping of the Voronoi boundary and the gradient map; finally, the compactness factor is weighted and corrected by using the topology index to obtain stemness maintenance confidence, and target coordinates are output. Through the dual verification of geometric topology and physical gradient, the application realizes the joint constraint screening of the features inside and outside the cells.

The invention discloses an osteoclast culture method as well as a culture medium and application thereof, the culture method is based on primary monocyte extraction, inoculation is performed according to a specific density, then quantitative M-CSF is added, and culture is performed overnight; on the second day, when the cell confluence degree of the mononuclear bone marrow hematopoietic stem cells is 5-30%, supplementing quantitative RANKL into each hole, continuously culturing for 2-4 days, then changing the solution, and obtaining mature osteoclasts on the sixth-seventh day. According to the invention, the optimization relationship among the cell activity, the cell density and the culture medium in the primary culture process of the osteoclast is deeply studied, and from the perspective of cost reduction and efficiency improvement, the use cost of cell factors is remarkably reduced, the number of times of liquid change is reduced, and the traditional culture time of 10-12 days (calculated from cell extraction) is shortened to 6-7 days. The average diameter of the mature osteoclast cultured by the method is larger, the number of fused cell nucleuses is larger, the space proportion of the mature osteoclast is not less than 80%, and the cell quality is obviously higher than that in the prior art.

The application discloses a molecular marker for identifying pepper CMS lines and maintainer lines and application thereof. The molecular marker for identifying pepper CMS lines and maintainer lines is that a "TTATAATAATG" fragment deletion exists at 120,582 bp of a mitochondrial genome of a pepper CMS line 'BA3'; and the pepper mitochondrial genome is subjected to PCR amplification by using the upstream and downstream primers of the molecular marker, so that the pepper CMS lines and the maintainer lines can be identified and distinguished. The application can provide a reference for identifying a cytoplasm type of the pepper, identifying a male sterile type of the pepper, efficiently selecting and utilizing the pepper CMS lines, and has a wide application prospect in pepper breeding practice.

This disclosure provides an E. coli strain, which lacks thioredoxin reductase activity encoded by trxB and thioredoxin 1 activity encoded by trxA, and glutathione reductase activity encoded by gor. Said E. coli strain expresses a mutated AhpC protein having glutathione reductase activity and a cytosolic prokaryotic disulfide isomerase. The E. coli strain has an oxidative cytosol and can be used to efficiently produce proteins having disulfide bonds.

Tumor glycocalyx may be regarded as a glycan “canopy” above the tumor plasma membrane. Disclosed are methods of treating a neoplastic malignant cell cancer wherein the neoplastic malignant cancer cell has a malignant glycocalyx B. Also disclosed are methods of inducing apoptosis of one or more neoplastic malignant cells and methods of increasing a subject's responsiveness to an immunotherapy wherein the neoplastic malignant cancer cell has a malignant glycocalyx B.

Due to the constituent components of the cell walls of filamentous fungi, the texture when eating a filamentous fungus body resembles a paper-like texture, and applications other than meat analogs are limited. Additionally, filamentous fungus bodies have characteristics of absorbing water but also readily releasing water, and having no binding property between mycelia, and thus even when a crushing treatment is performed, it has been difficult to process the result into foodstuffs that have a feeling of unity due only to a filamentous fungus culture product, or into foodstuffs that have uniform and smooth paste-like physical properties. This edible filamentous fungus body having an adjusted texture can be provided by treating a filamentous fungus body with a cell wall degrading enzyme.

The invention belongs to the cross technical field of cell engineering, nano-drugs and biological manufacturing, and particularly discloses a drug-loaded vesicle based on denucleated cells as well as a preparation method and application of the drug-loaded vesicle. According to the invention, a cell suspension and a cytoskeleton relaxant are co-incubated to relax an actin skeleton and weaken nucleoplasm connection; then loading the treated cell suspension on a multi-layer discontinuous density gradient centrifugal medium, realizing physical separation of cell nucleuses and cytoplasm through high-speed centrifugation according to buoyancy density difference, and collecting components of a specific interface to obtain high-purity denucleated cells with a complete membrane structure; then co-incubating the denucleated cells and ROS response type lipidosome (co-carrying therapeutic siRNA and a sound-sensitive agent Ce6) prepared in advance, so that the lipidosome is wrapped or anchored by a denucleated cell membrane; finally, the composite system is subjected to extrusion treatment through a microporous membrane, the drug-loaded vesicles uniform in particle size and stable in structure are obtained, and the drug-loaded vesicles are suitable for various application scenes such as anti-tumor treatment and RNA vaccine delivery.

The invention discloses an ingenol ABC transport protein ElABCG39 as well as a coding gene and application thereof. The protein is positioned in a cytoplasmic membrane, and expression of the protein in yeast proves that the protein has ingenol transmembrane efflux transport function, improves the ingenol tolerance of the yeast, and proves that the ElABCG39 has certain substrate specificity in the yeast. The ingenol ABC transporter ElABCG39 can be used for producing ingenol by fermentation of microorganisms or plant tissues.

The application discloses an exosome-based protein delivery system and application thereof. The exosome-based protein delivery system comprises a loading subsystem and / or an unloading subsystem, the polypeptide peptide segment expressed by the nucleotide sequence of the loading subsystem comprises CD63, a connecting peptide, an enterokinase cleavage site and a delivered protein or polypeptide, and the polypeptide peptide segment expressed by the nucleotide sequence of the unloading subsystem comprises CD63, a connecting peptide, enterokinase or enterokinase light chain. The application also constructs a human umbilical cord mesenchymal stem cell strain hUC-MSC CDNF‑exo on November 13, 2024 in the Guangdong Provincial Microbial Culture Collection Center, and the preservation number is GDMCC NO:65478. The human umbilical cord mesenchymal stem cell strain hUC-MSC CDNF‑exo The purpose of delivering CDNF into the cytoplasm of target cells is achieved.

The invention discloses application of inhibiting combination of Hic-5 and SMAD7 and / or inhibiting Hic-5 in treatment of pulmonary arterial hypertension, and finds that PLEKHH2 and SMAD7 have competitive combination with Hic-5 protein in lung tissue, more SMAD7 can be released into cytoplasm by preventing combination of Hic-5 and SMAD7, and SMAD2 / 3 phosphorylation is inhibited for the first time, so that the purpose of treating the pulmonary arterial hypertension is achieved. The invention provides a new research and development direction for the development of pulmonary arterial hypertension treatment medicines.

The invention discloses application of oxygen-carrying protein VHb in construction of high-yield astaxanthin engineering bacteria, and belongs to the technical field of biology. According to the method, carRPY27R, carB, fusion genes crtW-crtZ and GGPPS are iteratively integrated onto a yarrowia lipolytica chassis genome by utilizing a synthetic biological technology, so that two recombinant strains for producing astaxanthin are obtained. On the basis, the oxygen-carrying protein VHb from vitreoscilla is expressed, and compared with the oxygen-carrying protein VHb which is not expressed, the VHb is positioned to the endoplasmic reticulum, so that the accumulation level of astaxanthin is improved by 272.99%, and the biomass is improved by 94.15%. According to the method, the oxygen-carrying protein VHb is expressed on cytoplasm and organelle, so that the synthesis efficiency and biomass of astaxanthin are remarkably improved. In addition, the intracellular dissolved oxygen level is improved, so that the cost for improving the dissolved oxygen in the fermentation process is reduced.

It was discovered herein that neither the nuclear localization signal (NLS) nor the prion-like domain (PLD) of TDP-43 are required for in vitro embryonic stem cell culture and differentiation into motor neurons. ES cells expressing these TDP-43 mutants and differentiated into motor neurons that exhibit an ALS-like phenotype, from which the TDP-43 mutants redistribute to and accumulate in the cytoplasm, and the inability to regulate cryptic exon splicing, such that these cells can serve as a model for TDP-43 proteinopathies for testing candidate therapeutics that can dissipate such proteinopathies. In addition, these ES cells can be used to successfully generate non-human animals, e.g., mice, that also exhibit hallmark symptoms of ALS and can be used to test candidate agents useful for treating TDP-43 proteinopathies.

The application discloses an animal cell membrane protein and cytoplasm protein separation and extraction reagent and application, and belongs to the technical field of protein separation and extraction. The membrane protein and cytoplasm protein separation and extraction reagent provided by the application can realize separation and extraction of animal cell membrane protein and cytoplasm protein without using complex methods such as repeated freezing and thawing of liquid nitrogen to lyse cells, is simple to operate, efficient and convenient, has high extraction efficiency on membrane protein, and can effectively reduce cross contamination of cytoplasm protein in cell membrane components.

This invention discloses a method for studying the mechanism by which the mucoprotein channel 1 (TRPML1) regulates the outflow of zinc ions from lysosomes and induces changes in mitochondrial ATP. This method uses 13 nm AuNPs as loading materials to construct a logic-gated sensor for pH and zinc ion regulation within lysosomes and for TRPML1 mRNA in the cytoplasm. It also uses 5 nm AuNPs as loading materials to construct a sensor targeting mitochondrial ATP for cell imaging. This method enables the detection of mRNA and zinc ions in the cytoplasm, lysosomes, and mitochondria. 2+ ATP-associated ATP sensors, Zn 2+ Three sensors—a sensor, an mRNA sensor, and an X-ray sensor—are used simultaneously to image cells, aiming to answer questions about the mechanisms of ATP changes in mitochondria under the regulation of zinc ion efflux from lysosomes by TRPML1.