43 results about "Nuclear membrane" patented technology

The invention belongs to the field of nano preparations, and discloses a preparation method of nano microspheres coated with a nicotinamide mononucleotide shell membrane structure, which comprises the following steps: coating NMN in a pregel formed by chitosan and sodium tripolyphosphate, heating and stirring, adjusting the pH value to separate out chitosan-coated nano microspheres, drying to remove moisture on the surfaces of the nano microspheres, and drying to obtain the nano microspheres coated with the nicotinamide mononucleotide shell membrane structure. Then selecting a water phase and an oil phase in a proper proportion, preparing a chitosan / sodium alginate composite nano-microsphere with sodium alginate as an outer membrane by adopting an emulsion cross-linking method, and washing and drying to obtain the NMN nano-microsphere with a core-shell structure. According to the invention, NMN is coated in the nano-microsphere which is composed of two materials and has a nuclear membrane structure, so that the stability of NMN in a natural environment can be improved, hydrolysis of NMN by gastric acid and gastrointestinal tract enzyme in a drug delivery process can be prevented, the bioavailability of NMN is improved, and a drug sustained release effect is achieved.

The invention provides a universal method for preparing animal tissue mononuclear suspension. A universal solution is provided for preparation of animal tissue single cell nucleuses, the method is suitable for different tissue types of various species, and the problem of unstable extraction caused by tissue differences in a traditional method is solved. Specifically, through multi-stage grinding and three dynamic quality inspection feedback mechanisms, hardness and cracking difficulty of different tissues are adapted, redundant operation is reduced to avoid damage of nuclear membranes caused by excessive grinding or excessive purification, nuclear membrane integrity and RNA stability are ensured, and extracted cell nucleuses are good in state and relatively high in integrity and can meet the requirement of single cell sequencing.

The invention provides a lysis solution for rapidly extracting animal and plant genomes and an application method thereof.The lysis solution is prepared from, by mass, 20-25 parts of Tris-Hcl, 10-15 parts of LICI, 5-10 parts of EDTA, 1-3 parts of 3-sulfopropyl hexadecyl dimethyl betaine, 1-3 parts of (NH4) 2SO4 and 30-40 parts of ddH2O. The 3-sulfopropyl hexadecyl dimethyl betaine and (NH4) 2SO4 in the formula of the lysis solution can effectively destroy cell membranes and nuclear membranes; the lysate prepared by the method is clear in component and simple to prepare, DNA precipitation or column purification can be directly carried out after lysis, meanwhile, the lysate is suitable for various sample types including animal tissues and plant tissues, a formula does not need to be adjusted according to different sample types, and compared with a traditional lysate, the lysate has the advantages that the cost is low, and the efficiency is high. The optimized formula has the advantages of high splitting efficiency, wide application range, high DNA purity, high extraction speed and good DNA integrity.

The invention discloses a method for promoting ovulation of sexual maturity zebrafish by using Gap27, and relates to the technical field of artificial breeding of fish, a gap connecting protein inhibitor Gap27 is applied to ovulation induction of zebrafish, an intercellular communication network in follicles is accurately intervened, the problem of asynchronous ovulation is solved, and in the final stage of maturation of oocytes, ovulation of sexual maturity zebrafish is promoted. Close substance and information exchange is carried out among follicular granule cells through a gap connecting channel formed by connecting protein Cx43, and Gap27 is used as a specific peptide fragment, can simulate an extracellular ring structure of Cx43, and competitively binds with Cx43 on an adjacent cell membrane, so that formation and functions of the gap connecting channel are physically blocked, and the follicular granule cells are separated from each other. When ovarian tissues of sexually mature female fish are exposed in a Gap27 solution, the transmission of inhibitory signals from granular cells to oocytes can be effectively interrupted, the rupture of nuclear membranes of the cells is promoted, and the final maturation process is completed.

This invention discloses a kit and method for extracting cell nuclei from plant tissues, belonging to the field of biotechnology. The kit includes a nuclear lysis buffer and a nuclear washing buffer. The nuclear lysis buffer is prepared using nuclease-free water with sucrose, Tris-HCl (pH 7-8), NaCl, MgCl2, a surfactant, DTT, an RNase inhibitor, and spermidine. The nuclear washing buffer is prepared using nuclease-free water with sucrose, Tris-HCl (pH 7-8), NaCl, MgCl2, DTT, an RNase inhibitor, and spermidine. This kit is suitable for extracting cell nuclei from fresh yew stem tissue, yielding cell nuclei with intact nuclear membranes, stable gene expression, high purity, and few impurities, suitable for single-cell sequencing.

The invention belongs to the technical field of animal reproductive biology and cell engineering, and particularly relates to a fluorescence labeling expression system for visualizing an oocyte nucleus maturation process and application of the fluorescence labeling expression system. The fluorescence labeling expression system comprises an eukaryotic expression vector pVenus-LMNB1; the eukaryotic expression vector pVenus-LMNB1 comprises a nucleotide sequence for coding the fusion protein Venus-LMNB1. According to the invention, a nuclear fiber layer protein stably expressed on a nuclear membrane is selected as a research object, a eukaryotic expression vector pVenus-LMNB1 is constructed by using a molecular cloning technology, an LMNB1 gene containing a green fluorescent label Venus is overexpressed in a porcine oocyte by using a microinjection technology, fluorescent protein labeling of a structural protein on the nuclear membrane is utilized, and the expression vector pVenus-LMNB1 is constructed. The expression positioning condition of the LMNB1 in the maturation process of the porcine oocytes is observed, and a powerful tool is provided for development of in-vitro culture related researches of the porcine oocytes.

This invention discloses a method for constructing a tomato core membrane-associated protein interaction library based on Gateway technology under Fusarium oxysporum infection conditions and its application, relating to the field of genetic engineering technology. The method includes: using tomato root tissue infected with Fusarium oxysporum as material, extracting total RNA and enriching mRNA; synthesizing first-strand cDNA using Biotin-attB2-Oligo(dT) primers; synthesizing double-stranded cDNA enzymatically; ligating it with a specific double-stranded attB1 adapter; introducing attB1 and attB2 sites at both ends of the cDNA; cloning the cDNA into the donor vector pDONR222 via BP recombination to construct a primary entry library; and recombinating the primary entry library with vectors pGADT7 and pPR3-N via LR recombination to construct a nuclear secondary library and a membrane secondary library, respectively. The constructed libraries have high titers, long insert fragments, and low empty vector rates. This invention also relates to the application of the library in screening the interaction between Fusarium oxysporum effector proteins and tomato nuclear membrane-associated proteins, providing an efficient and specific resource platform for studying plant-pathogen interaction mechanisms.

The present application relates to the technical field of digital pathology image analysis, and discloses an animal tissue pathological section cell atypia image automatic grading method and system, the method comprises the following steps: scanning the whole section image to obtain a high-power field image by traversing the tumor area with a sliding window; inputting a star-convex polygon regression kernel instance segmentation network trained by multi-species animal tumor annotation data to obtain each nuclear boundary; extracting morphological parameters such as nuclear area, long-short axis ratio, nuclear-plasma ratio, nuclear membrane regularity and chromatin texture; detecting and counting nuclear mitotic figures; introducing a species adaptive baseline correction to normalize the morphological parameters and then inputting the normalized morphological parameters into a classifier to output a three-level atypia grading.

The present application relates to the field of tumor detection, and in particular to a biological sample analysis method and system based on nuclear pore distribution patterns. The method provided by the present application comprises: S101, obtaining microscopic image data of a biological sample; S102, obtaining a first region of interest, a second region of interest and a third region of interest; S103, drawing a nuclear membrane edge curve; S104, extracting nuclear pore distribution-nuclear membrane curvature distribution data of cells in the biological sample; S105, identifying the nuclear pore distribution pattern of the cells in the biological sample; S106, evaluating the state of the tissue or organ corresponding to the biological sample. Based on the identified nuclear pore distribution pattern of individual cells in the biological sample, the present application can evaluate the state of the tissue or organ corresponding to the biological sample according to the overall nuclear pore distribution pattern of the cells in the biological sample, such as whether it is a tumor sample, whether it has a tendency to cancer, etc., which is of great significance for early diagnosis of tumors.

The invention provides an application of a lipid peroxidation inhibitor in preparation of an anti-premature senescence syndrome drug or an anti-aging drug. Experiments prove that by adding the lipid peroxidation inhibitor, the cell nucleus morphology of cells can be effectively maintained, the expression of intracellular senescence-related beta-galactosidase is reduced, the senescence characteristic of the cells is delayed, and nuclear membranes and nuclear fiber layers damaged due to Progyrin accumulation are repaired, so that the normal functions of the cell nucleuses are recovered, the activity and functions of the cells are improved, and the anti-aging effect is achieved. Powerful cellular level support is provided for relieving and treating the early senescence syndrome and senescence.

The present application relates to the technical field of edge detection algorithm, and particularly relates to an end-to-end cell aging state prediction method based on nuclear membrane morphology, which comprises the following steps: step one, collecting a fluorescence image of human fibroblasts subjected to immunofluorescence staining through a fluorescence microscope; step two, performing data enhancement processing on nuclear membrane data, amplifying training data, and improving model generalization capability; step three, generating a prediction of nuclear membrane segmentation mask through a deep neural network, and obtaining mask information; and step four, simultaneously inputting the mask information into an ellipticity module, a smoothness module and a concave degree module for multi-angle calculation and evaluation. The present application evaluates nuclear membrane morphology changes from multiple angles to meet the growing demand for quantification of cell aging state in the field.

The present disclosure provides nucleic acid sequences and nucleic acid delivery constructs comprising “zip code” sequence(s) that home, target, cross a cytoplasm, and / or cross a nuclear membrane of a target cell (e.g., a diseased cell such as a cancer cell) or cell population (e.g., tissue), and integration sequence(s) that allow for integration of at least a portion of such nucleic acid or nucleic acid delivery system into a genome of such target cell. The present disclosure also provides non-naturally occurring nucleic acid constructs and delivery systems comprising such Zip Code and integration sequences as well as one or more cargo molecules that may be coupled covalently or non-covalently to such nucleic acid constructs and systems. Further provided herein are methods of diagnosing and treating diseases such as cancer using the target cell specific nucleic acid constructs and systems described herein.

The present invention provides a composition containing polymerase-labeled polyantibodies and polymerase-labeled anti-nanoantibodies, wherein polymerase-labeled antibody compositions of different sizes are matched to different localized antigens, thereby improving the penetration of secondary antibodies into cell nuclear membranes and overcoming the problems of unevenness and low universality exhibited by different types of antigen detection; and polyethylene glycol-connected polyethyleneimine compounds are selected as skeleton materials, and by optimizing the selection of skeleton materials, the use of highly toxic raw materials is avoided, the safety of product production is improved, the hydrophilicity of polymerase-labeled antibodies is improved, background staining is effectively reduced, and the volume of immunohistochemical secondary antibodies is reduced, thereby further improving their molecular mobility and penetration capabilities.

The invention discloses a novel grape fruit cell nucleus extraction method for single cell sequencing, and belongs to the technical field of cell nucleus extraction. The invention provides a method for extracting cell nucleuses of grape fruits. The method comprises the following steps: washing grape pulp tissues, carrying out light-proof enzymolysis to obtain protoplast, splitting the protoplast to obtain cell nucleuses, carrying out quality inspection on the cell nucleuses, removing impurities and the like. According to the method, the washing step is added, so that sugar impurities in the extracting solution are effectively reduced. Compared with direct nucleus extraction, the method has the advantages that the extraction process is milder, the extraction time is short, a large number of nucleuses with normal forms, high nuclear membrane integrity and few background fragments can be obtained, the method can be used for subsequent single-cell nucleus sequencing, and the sequencing result is good.

The invention relates to a Pin1 protein targeted antagonistic polypeptide, a targeted antagonistic polypeptide nano-micelle as well as a preparation method and application of the targeted antagonistic polypeptide nano-micelle. The Pin1 protein targeted antagonistic polypeptide comprises a targeted antagonistic polypeptide and a cell-penetrating peptide which are connected in sequence, the amino acid sequence of the targeted antagonistic polypeptide comprises any one of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4 or SEQ ID NO: 5; the amino acid sequence of the cell-penetrating peptide comprises SEQ ID NO: 6. The Pin1 protein targeted antagonistic polypeptide can efficiently cross cell membranes and nuclear membranes, is specifically combined with the Pin1 protein in human metastatic pancreatic adenocarcinoma cells, antagonizes the biological function of the Pin1 protein and kills tumor cells. Meanwhile, the Pin1 protein targeting antagonistic polypeptide is combined with gemcitabine, so that the sensitivity of a mixed in-situ tumor model of human metastatic pancreatic adenocarcinoma cells and cancer-related fibroblasts to gemcitabine can be enhanced, the anti-cancer effect of a single drug is amplified, and the anti-tumor effect of 1 + 1 > 2 is generated.

To provide methods of reacting a cell nucleus informational molecule with an analytical biomolecule.SOLUTION: The invention relates to a system and methods of enhancing access to nuclear informational molecules, such as DNA, RNA, and proteins, by analytical biomolecules, such as transposome complexes, by treating nuclei with a nuclear permeability enhancer, and to methods of using nuclear membranes, cell membranes, and external compartmentalization approaches as contiguity preserving elements. The present invention provides, for example, a method of reacting a cell nucleus informational molecule with an analytical biomolecule, comprising contacting the cell nucleus with a nuclear permeability enhancer and reacting the informational molecule with the analytical biomolecule.SELECTED DRAWING: None

The invention provides a cell virtual staining method and system, computer equipment and a storage medium, and belongs to the field of cell detection.The method comprises the steps that a microscopic image of a to-be-stained cell is obtained; a multi-scale structural feature map is extracted from the microscopic image, the multi-scale structural feature map is weighted based on a regional attention mechanism to obtain cell structural features of the cell to be stained, and the weight of the boundary features is in positive correlation with the cell nucleus density; extracting color distribution features from a preset reference dyed image, and performing spatial alignment and semantic mapping on the color distribution features and the cell structure features to obtain fusion features; and performing image reconstruction according to the fusion features to obtain a staining image of the to-be-stained cell. According to the method, the display effect on fine structures such as cell nuclear membranes and chromatin distribution is improved, and the high-precision requirement of clinical diagnosis on cell substructure recognition is met.

The present disclosure describes a method of treating a dsbNETs-associated pathology or con- dition in a subject in need thereof, comprising administering a therapeutically effective amount of a dsbNETs modulator in the subject, wherein dsbNETs repair or misrepair damaged DNA or wherein excessively induced or persistent dsbNETs trigger DNA damage.

Alternative chromatin mapping methods, other than ChIP, typically map a chromatin associated biomolecule by successive binding of a specific antibody in situ in permeabilized nuclei, and then tethering chromatin-cleaving or -modifying enzymes. These methods are however limited by their requirement for a nuclear envelope to hold the chromatin together during wash steps and buffer exchanges. This results in large input material requirements due to sample losses, limitations in what types of samples can be processed, increased variability due to handling steps, and an inability to study chromatin outside of its compacted form in the nucleus. A new method for preparing chromatin from cells for downstream genomic chromatin mapping is disclosed. The method uses dilution, precipitation, and dialysis to replace wash steps and buffer exchanges, allowing the entire protocol to take place in a single tube with no removal of material prior to DNA extraction.

The invention relates to the technical field of plant genetic engineering, in particular to application of a millet transcription factor S1Fa in regulation and control of plant stress resistance. The invention provides a transmembrane transcription factor SiS1Fa which has a CDS area length of 243 bp and is positioned in a cell membrane and a nuclear membrane, and plays an important role in the salt resistance process of millet. The millet plant with overexpressed SiS1Fa shows better growth condition and biomass under salt stress treatment, is obviously superior to WT, and particularly shows prominent ROS (reactive oxygen species) removal capability and oxidative damage relief.

This invention discloses a method for constructing a cell nucleus model based on Abaqus-Python, relating to the field of cell stress analysis technology. This method utilizes Abaqus finite element software to draw a cell nucleus model and analyze its mechanical properties. Simultaneously, it employs Python for secondary development, simplifying the model drawing process through scripting and improving the efficiency and accuracy of model creation. This model can be used to calculate the stress distribution of the cell nucleus's main structures—the nuclear membrane and nuclear lamina—during the process of nanoneedles penetrating the cell nucleus, and to optimize nanoneedle parameters by analyzing the stress concentration of the nuclear membrane during penetration.

The invention provides a detection method for realizing visual protein-protein interaction by co-transferring an outer nuclear membrane. The method comprises the following steps: constructing an expression vector of an outer nuclear membrane localization protein and a fluorescent protein; recombining the expression fragment of the bait protein with an expression vector to construct a bait protein expression vector; transforming and extracting plasmids; constructing a prey protein expression vector; transforming and extracting plasmids; transfecting bait protein expression vector plasmids and prey protein expression vector plasmids into cells in proportion, and culturing; and performing fluorescence imaging. And the co-localization condition of the protein on the outer nuclear membrane is directly observed by utilizing fluorescence labeling, so that the interaction of the protein can be judged. Positioning is clear and visual; and the method has high specificity, living cell compatibility and high throughput potential, so that the method has a wide application prospect in protein function research and drug screening.

The present application relates to the technical field of genetic engineering, and particularly relates to a screening method of an anti-aging drug. The screening method comprises: treating cells overexpressing GFP-progerin protein with a drug to be screened, and comparing the cells overexpressing GFP-progerin protein without treatment of the drug to be screened; and judging the anti-aging effect of the drug to be screened according to the change of cell morphology. The present application finds that progerin protein positioned on the nuclear membrane can make cells show obvious aging characteristics, including nuclear membrane shrinkage and nuclear membrane budding. Progerin protein positioned in the nucleus does not make cells show obvious aging characteristics. When a drug has an anti-aging effect, progerin protein positioned on the nuclear membrane will be transferred to the nucleus. The present application provides a screening method of an anti-aging drug based on the above, which can accurately and efficiently screen an anti-aging drug.

The invention provides a FISH (fluorescence in situ hybridization) probe for detecting a new coronavirus Armike strain as well as preparation and application of the FISH probe. The FISH probe comprises three steps of determining a CDC-O target area, designing and synthesizing an oligonucleotide library, amplifying a probe template library and preparing the fluorescent probe. The prepared FISH probe is uniform and controllable in size, the size ranges from 20 bp to 100 bp, and the FISH probe can easily penetrate through a cell nuclear membrane, so that the hybridization efficiency is improved; meanwhile, the preparation method is not limited by BAC cloning, and only one-time chemical synthesis is needed, so that the synthesis cost is reduced, and the batch difference caused by repeated synthesis can be reduced. The probe can be used for in-vitro qualitative detection of the Obemitra virulent strain in a new coronavirus patient tissue slice soaked by formalin or embedded by fixed paraffin, so as to achieve the purpose of accurate diagnosis.

Disclosed are systems, devices, and methods for delivering substances from the cytoplasm into the nucleus of cell by selectively disrupting the nuclear membrane. In some aspects, a method for delivering substances into a nucleus of a cell includes creating one or more pores in the nucleus of the cell by imposing one or more nanoscale geometric and / or physical constraints on the cell. In some embodiments of the method, the method includes enabling entry of substances into the nucleus through the created one or more pores.

The invention discloses a kit for extracting plant tissue cell nucleuses, and belongs to the technical field of biology. The kit comprises a cell nucleus lysis solution and a cell nucleus cleaning solution, the cell nucleus lysis solution is prepared from cane sugar, Tris-HCl with the pH value of 7-8, NaCl, MgCl2, a surfactant, DTT, RNase inhibitor and spermidine by using nuclease-free water, and the cell nucleus cleaning solution is prepared from sucrose, Tris-HCl with the pH value of 7-8, NaCl, MgCl2, a surfactant, DTT, RNase inhibitor and spermidine by using nuclease-free water. The cell nucleus cleaning solution is prepared from sucrose, Tris-HCl, NaCl, MgCl2, DTT (dithiothreitol), RNase inhibitor and spermidine by using nuclease-free water, wherein the pH (Potential of Hydrogen) of the Tris-HCl is 7-8. The kit is suitable for cell nucleus extraction of fresh tissues of taxus chinensis stems, the obtained cell nucleus is complete in nuclear membrane, stable in gene expression, high in purity and few in impurity, and the kit can be used for single cell sequencing.

The invention discloses a nucleic acid delivery system and a preparation method and application thereof, and belongs to the technical field of biological materials. According to the invention, a benzenesulfonamide functional group is introduced into a GSs molecular skeleton, such that a novel benzenesulfonamide functionalized gemini surfactant (NGS) is constructed. Wherein benzenesulfonamide is a specific endoplasmic reticulum targeting group, and can realize endoplasmic reticulum targeting enrichment through specific binding with a sulfonylurea receptor on an endoplasmic reticulum membrane, so that the NGS gene delivery vector is distributed towards the endoplasmic reticulum. After the NGS and the pDNA form a compound, the compound enters cells through cavette protein mediated endocytosis, then targets the endoplasmic reticulum and releases the pDNA, the pDNA is delivered into the cell nucleus by utilizing the close relation between the endoplasmic reticulum and the nuclear membrane, the gene transfection efficiency is finally improved, meanwhile, the biocompatibility is further improved by introducing the gamma-polyglutamic acid (gamma-PGA) with negative charges, and the application prospect is wide.

The invention relates to the technical field of biology, in particular to a preparation method of anisotropic microgel for regulating and controlling cell mechanics. According to the anisotropic microgel and the preparation method thereof, mechanical and biological signal crosstalk can be formed through long-axis polarity and cell membrane-intracellular skeleton-cell nucleus, the adhesion, proliferation and anti-apoptosis capabilities of nucleus pulposus cells are improved, the stability of the nuclear membrane is improved, DNA damage and leakage are reduced, cGAS-STING expression is down-regulated, the young state level of the loaded nucleus pulposus cells is maintained, and the anisotropic microgel can be used for preparing the medicine for treating the nucleus pulposus cells. Extracellular matrix metabolism unbalance in intervertebral disc degeneration is corrected, secretion of various inflammatory factors and protease is reduced, and the pathological process of intervertebral disc degeneration is delayed.

The invention relates to the technical field of biochemical sensing and fluorescence imaging, in particular to a chromazole near-infrared viscosity-sensitive probe, a preparation method thereof and application of the probe in living cell imaging. The preparation method of the probe comprises the following steps: dissolving tryptazole tetrafluoroborate and benzaldehyde or a derivative thereof in acetonitrile, stirring at room temperature to react, evaporating to remove a solvent after the reaction is finished, and separating and purifying through column chromatography to obtain the tryptazole near-infrared viscosity sensitive probe. The application of the probe in living cell nucleus imaging is to mark chromosomes in a cell mitosis process, so that chromosome migration behaviors are visualized. The probe is high in utilization rate, mild in preparation reaction condition, simple and convenient to operate, green and efficient. The cell nucleus of the living cell can be quickly dyed at low concentration, the cytoplasm, the nuclear membrane and the nucleolus of the living cell can be distinguished through the fluorescence brightness, and the mitosis process of the cell can be traced. A feasible strategy is provided for developing a near-infrared probe with a rapid cell nucleus staining function.

The invention discloses a cell nuclear membrane regularity calculation method and device, equipment and a storage medium, and relates to the field of image processing. Sampling and preprocessing the original cell nucleus image data to obtain a complex contour sequence; performing fast Fourier transform and spectral analysis on the contour data of the complex contour sequence, and calculating the frequency component of the complex contour; calculating nuclear membrane regularity index data according to the set Fourier transform frequency threshold; the nuclear membrane regularity index data comprises positive frequency nuclear membrane regularity, negative frequency nuclear membrane regularity and comprehensive nuclear membrane regularity; and performing quantitative evaluation on the regularity of the nuclear membrane based on the nuclear membrane regularity index data, and outputting an evaluation result. According to the scheme, accurate quantitative evaluation on the regularity of the nuclear membrane is realized through a Fourier analysis technology, the limitation of a traditional method is overcome, and powerful technical support is provided for cell pathology diagnosis.