1298 results about "Intracellular" patented technology

The invention discloses an indole-benzothiadiazole derivative as well as a preparation method and application thereof. The general structural formula of the indole-benzothiadiazole derivative provided by the invention is shown in the specification, wherein R is selected from one of methyl piperazinyl, diethylamino, azetidinyl and pyrrolidinyl. The derivative can be specifically combined and fluorescently labeled with mitochondrial RNA in cells, and has good selectivity, high fluorescence signal-to-noise ratio and excellent anti-interference capability in a complex biological system. Meanwhile, the indole-benzothiadiazole derivative provided by the invention is simple in synthetic route, easily available in raw materials, good in stability and convenient to store and use, can be specifically combined with RNA and generate strong fluorescence response, is suitable for real-time detection and imaging of in-vitro and in-vivo mitochondrial RNA, and is an efficient mitochondrial RNA micromolecular fluorescent probe.

The instant disclosure relates to lipid particles that harbor cationic lipids, the particles found to be capable of delivering associated cargoes - particularly nucleic acid cargoes when formulated as nucleic acid-lipid particles - intracellularly to skin tissue cells when administered topically to a subject. The instant disclosure provides compositions comprising such lipid particles, optionally in association with a therapeutic agent (e.g., a therapeutic mRNA and / or nucleic acid controller system), as well as methods and kits for delivering a lipid particle-associated therapeutic agent and / or for treating or preventing a disease or disorder, e.g., a skin disease or disorder, in a subject, using one or more lipid particle compositions provided herein.

The invention relates to the technical field of biomedicine, discloses mutant photo-induced ion channel ChR-2 protein, and further discloses a nucleic acid construct, an expression vector, a cell, related application and a computer model. The light-sensitive channel ChR2 protein mutant obtained by the invention has stronger light current, and the light-sensitive capability of the light-sensitive channel ChR2 protein mutant is at least improved by 100 times; besides, the invention also obtains a nucleotide sequence for coding the light-sensitive channel ChR2 protein mutant, constructs a recombinant expression vector, and obtains the light-sensitive channel ChR2 protein with higher expression quantity and stronger light sensitivity, and the light-sensitive channel ChR2 protein is very suitable for expression in cells of mammals (especially human); according to the invention, the rhodopsin in different channels is systematically studied by modifying the position G224 in the helix 6 of seven transmembrane helix motifs, which proves that the mutation of the position G224 in the helix 6 in WT ChR2 accelerates the photosensitivity of the channels, and the considered position is homologous in the helix 6 of the rhodopsin in different channels.

The application relates to an engineering bacterium for high yield of observation blue and a construction method and application thereof, and belongs to the technical field of biological synthesis of natural dyes. The engineering bacterium for high yield of observation blue expresses icd, bpsA, glnA Y405F and gdhA; the engineering bacterium knocks out acnR, yggB, glsK, aceA and ldh. By knocking out the yggB and aceA genes, performing site-directed mutation (Y405F) on the glnA gene, replacing the glsK gene with the glnA Y405F gene, replacing the acnR gene with the icd gene, replacing the ldh gene with the gdhA gene, and integrating the bpsA gene, the application can block the formation of by-products such as lactate and succinic acid in the observation blue synthesis process, improve the flow direction of the citric acid->isocitric acid->alpha-ketoglutaric acid->glutamic acid->glutamine path, and then improve the intracellular glutamine concentration, so that the yield of observation blue is improved.

The invention belongs to the field of biological medicines, and relates to a drug-loaded nano-vesicle as well as a preparation method and application thereof. The drug-loaded nano-vesicle comprises a vesicle core and a drug-loaded nano-vesicle, wherein the vesicle core comprises siRNA (small interfering Ribonucleic Acid) capable of specifically targeting and silencing an NR1D1 gene; the vesicle membrane is formed by fusing an erythrocyte membrane, a macrophage membrane, cardiolipin, cholesterol and lecithin. The drug-loaded nano-vesicle can specifically target macrophages in a sepsis immunosuppression stage, has an intracellular response release function, recovers BMAL1 and IGF2BP2-ATP6V1B2 / ATP6V0c axis functions by inhibiting NR1D1 expression, reconstructs a macrophage phagocytosis function and lysosome-dependent bacterium removal capability, and can be used for preparing a drug-loaded nano-vesicle with a specific targeting function. The survival rate of sepsis immunosuppression model animals is obviously improved; and the bacterial load is reduced. Compared with a traditional electroporation method, the preparation method disclosed by the invention has the advantage that the encapsulation efficiency of siRNA is remarkably improved.

The invention belongs to the field of biological medicine, and particularly relates to phase separation polypeptide and a preparation method and application thereof. The structure of the phase separation polypeptide is shown as a formula (I). According to the polypeptide compound provided by the invention, liquid-liquid phase separation can be carried out to form condensed liquid drops, and DNA damage response proteins are enriched and separated in cells. The polypeptide compound disclosed by the invention is simple in synthesis method and good in biocompatibility, and liquid-liquid phase separation can be carried out; dNA damage response protein can be captured in cells, DNA repair is interfered, and the tumor radiotherapy effect is enhanced. Therefore, the polypeptide compound provided by the invention can be used for tumor radiotherapy, chemotherapy and other treatment methods.

An acellular product may be derived from human placenta and may be used in various scenarios for wound healing. Because the product may be acellular, the product may be processed for storage and transportation with minimal degradation. The product may include various scaffolding such as biomaterials or human tissue, and the scaffolding may be infused with various plasmas and agents. The cell-free treatment may maintain the biological activity of many therapeutic agents found within cells and may possess multiple structural components to support cellular attachment. The structural components or scaffolds may function as a reservoir of highly diffusible chemotactic and cellular-programming factors that may be useful to treat injury and disease.

A composition of matter includes an antisense phosphorodiamidate morpholino oligomer (MO) that includes an MO base sequence. The MO base sequence is arranged to bind a corresponding complementary base sequence of messenger RNA (mRNA) transcribed from one or more genes for the beta subunit of human chorionic gonadotropin (hCG-β). An inventive method, for suppressing intracellular synthesis of a beta subunit of human chorionic gonadotropin (hCG-β), includes introducing such an MO into one or more cells.

The present invention relates to an antibody or an antigen-binding fragment thereof binding to CLDN18.2, an antibody-drug conjugate comprising same, and a use of the antibody and the antibody-drug conjugate. An anti-CLDN18.2 monoclonal antibody according to the present invention comprises a fully human antibody sequence, thereby having low in vivo immunogenicity, and exhibits excellent antigen affinity and binding ability specific to a low expression to a high expression level of the CLDN18.2 protein. Thus, the antibody is expected to exhibit high specificity and safety as an antibody-based therapeutic agent such as in the form of a monoclonal antibody and / or an antigen-binding fragment (scFv), an antibody-drug conjugate (ADC), an immune cell engager, a chimeric antigen receptor (CAR), a multispecific antibody, and the like. In addition, the antibody according to the present invention may undergo cellular internalization, enables an anti-CLDN18.2 antibody-drug conjugate comprising said antibodies to be conveniently prepared, and has excellent yield and quality and thus is expected to be highly likely to be developed as a drug. A drug conjugate comprising the anti-CLDN18.2 antibody according to the present invention has excellent in vivo anticancer efficacy and has an expanded therapeutic index (TI) and thus is expected to be usefully employable for the treatment and / or prevention of cancer diseases expressing CLDN18.2 and related diseases.

The invention discloses an RNA (Ribonucleic Acid) binding protein enrichment analysis method based on single cell expression profile data, which comprises the following steps of: writing an interactive online Web application program by using Shiny, carrying a data visualization tool, and using an RBP (Reactive Binding Protein)-gene database as a database; the method is specially designed for single cell data, different single cell data can be integrated for enrichment analysis, and expression profiles and RBP dynamic changes in single cells can be effectively captured and analyzed; rBP enrichment analysis is customized for single cell data, a single group and multiple groups of single cell expression data are preprocessed, including normalization processing, and a statistical model is used to evaluate enrichment significance at each time point, so that the analysis accuracy and biological correlation are improved; a dynamic visualization tool is provided, the dynamic visualization tool comprises a customizable network diagram, an interactive bar diagram and detailed table display, and a user can adjust a plurality of parameters of a view according to needs and directly download images and data from an interface.

The invention provides a multi-focus excitation confocal fluorescence microscopic imaging flow cytometry screening method. The method is characterized in that a laser beam generates three laser beams through a spatial light modulator, and the three laser beams are coherently superposed in a focal plane of a microscope objective to generate a two-dimensional optical lattice light field. And after the other laser beam is shaped by the cylindrical lens, linear laser optical tweezers are realized through the microscope objective, cells to be detected in an imaging detection area of the micro-flow chip are captured, and accurate active optical control on the rotation angle of the cells to be detected is realized. In a cell rotation process, a two-dimensional optical lattice light field rapidly scans in a focal plane at different angles, and a fluorophore in the cell is excited to generate a fluorescence signal. The confocal pinhole array installed at the position conjugate with the focal plane eliminates background fluorescence, a high-spatial-resolution fluorescence image of a three-dimensional structure in the cell is obtained, and cell screening is achieved. The method provided by the invention has the characteristics of high detection rate, high accuracy and the like, and has wide application prospects in the research fields of biology, medicine and life science.

The invention provides an experimental dyeing result image processing and calculating method. The method comprises the following steps: identifying a dyeing positive region in a preprocessed tissue sample digital image by using a first identification model, and carrying out area quantification on the identified dyeing positive region to obtain positive region area data; using a second recognition model to recognize and extract the intracellular cavitation region in the preprocessed tissue sample digital image, and performing area quantification on the extracted intracellular cavitation region to obtain cavitation region area data; calculating a dyeing positive region area proportion based on the positive region area data, and calculating a cavitation region area proportion based on the cavitation region area data; and inputting the area proportion of the dyeing positive region and the area proportion of the cavitation region into a preset disease evaluation model, and outputting a disease state evaluation result. According to the method, the accuracy and comprehensiveness of experimental dyeing image analysis are improved, and the automation degree and result credibility of pathological analysis can be remarkably improved.

The present invention relates to intracellular copper zinc superoxide dismutase inhibitors for protecting a subject from damage caused by radiation outbreak. The invention also relates to kits comprising SOD1 inhibitors and radiosensitizers for cancer cells, to the in vitro use of SOD1 inhibitors for protecting non-cancer cells from ionizing radiation, and to methods, compositions and uses related thereto.

The present invention relates to an isolated cellular targeted delivery system comprising a CD45+ leukocyte cell comprising within said cell a complex of one or more iron binding proteins and / or a label as well as methods for producing such isolated cellular targeted delivery system and uses of such system for therapy diagnosis and in particular for diagnosis of cancer, particularly metastatic cancer, in particular for therapy of cancer.

The present application relates to a kind of purine derivatives as the use of NSD2 and NSD3 inhibitor, the structural formula of the derivative is as shown in formula (I) or formula (II).The compound shows good NSD2, NSD3 inhibitory activity, can effectively reduce the methylation level of H3K36 in cell, and cause the reduction of cell proliferation activity of NSD2, NSD3 high expression tumor cell strain.

The present invention generally relates to systems and methods for imaging or determining nucleic acids, for instance, within cells. In some embodiments, the transcriptome of a cell may be determined. Certain embodiments are directed to determining nucleic acids, such as mRNA, within cells at relatively high resolutions. In some embodiments, a plurality of nucleic acid probes may be applied to a sample, and their binding within the sample determined, e.g., using fluorescence, to determine locations of the nucleic acid probes within the sample. In some embodiments, codewords may be based on the binding of the plurality of nucleic acid probes, and in some cases, the codewords may define an error-correcting code to reduce or prevent misidentification of the nucleic acids. In certain cases, a relatively large number of different targets may be identified using a relatively small number of labels, e.g., by using various combinatorial approaches.

The invention discloses a double-targeting probe and a preparation method and application thereof.The structure of the double-targeting probe is shown in the formula I. The double-targeting probe has the beneficial effects that SO2 can be monitored in the cell apoptosis process, the dynamic change of SO2 between mitochondria and lipid droplets in cells can be tracked, the core structure of the probe MLR is based on a coumarin part, and the core structure of the probe MLR can be used for detecting the apoptosis of the cells. The probe MLR is connected with a benzopyran cation structure specially designed for detecting SO2, SO2 generated in the death process of copper enables charges of a cation part in the probe MLR to be redistributed, a Michael addition reaction of SO2 and carbon-carbon double bonds is triggered, a fluorescence signal of the probe is changed, and the fluorescence signal is detected. The design of the probe provides a new tool for monitoring key biochemical changes in the copper death process, and is helpful for deeply understanding the molecular mechanism of copper death.

The invention relates to the technical field of biomedicine, in particular to a method for regulating and controlling the liquid-liquid phase separation capacity and function of DUX4 and application of the method, and the liquid-liquid phase separation capacity and function of the DUX4 are regulated and controlled by mutating an internal disordered region of the DUX4. The invention finds that the mutation has a remarkable inhibition effect on expression of DUX4 activated totipotent genes and FSHD related genes; according to the method, an intracellular liquid-liquid phase separation model is constructed through intracellular overexpression of DUX4; a research basis and a model are provided for subsequent LLPS research of DUX4, and thinking and mechanism explanation are provided for research of DUX4-mediated facial shoulder brachial muscular dystrophy (FSHD) and development of treatment targets.

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 an alkyl ether ester ionizable lipid compound and application thereof. The alkyl ether ionizable lipid compound is composed of a head group R1, a linker L and a tail group R2 in sequence. According to the alkyl ether ionizable lipid disclosed by the invention, two tertiary amine centers are connected by adopting a cyclic carbon chain structure, so that the delivery effect is obviously higher than that of ionizable lipid linearly connected with the two tertiary amine centers, and by adding an ether bond-containing tail structure in the alkyl ether ionizable lipid, the cell endocytosis and the cell endosome escape effect of LNP are improved; therefore, more mRNA is delivered into a living body and translated and expressed into corresponding proteins. Therefore, compared with the ionizable lipid in the prior art, the alkyl ether ionizable lipid disclosed by the invention has a better delivery effect.

The invention belongs to the technical field of biological image analysis and cell mechanics research, and particularly relates to a cell intracellular mechanical response visual analysis method based on image feature extraction, which comprises the following steps: acquiring original image data including a dynamic change process of cells under mechanical stimulation; performing structured segmentation on the original image data; extracting morphological features and dynamic behavior features of each data unit to obtain feature elements; classifying the feature elements according to a preset biological significance or mechanical response mode to form a plurality of feature element groups; and based on the classified feature element group, constructing a mapping relationship between the cell local region or the time slice and the mechanical response. According to the invention, the mechanical response of cells in a local area and a specific time window under mechanical stimulation can be analyzed, so that an analysis basis is provided for cell mechanical mechanism research, differentiation process regulation and control, drug effect evaluation and the like.

The invention relates to a sample pretreatment method for desorption electrospray ionization mass spectrometry imaging of a biological tissue frozen section, and the method comprises the following steps: pre-spraying the surface of the frozen biological tissue section, a pre-spraying solvent being a histocompatible solvent, and being used for enhancing the extraction of components in biological tissue cells, the detection sensitivity of components in biological tissues, especially in cells, is improved.

The present invention generally relates to systems and methods for imaging or determining nucleic acids, for instance, within cells. In some embodiments, the transcriptome of a cell may be determined. Certain embodiments are directed to determining nucleic acids, such as mRNA, within cells at relatively high resolutions. In some embodiments, a plurality of nucleic acid probes may be applied to a sample, and their binding within the sample determined, e.g., using fluorescence, to determine locations of the nucleic acid probes within the sample. In some embodiments, codewords may be based on the binding of the plurality of nucleic acid probes, and in some cases, the codewords may define an error-correcting code to reduce or prevent misidentification of the nucleic acids. In certain cases, a relatively large number of different targets may be identified using a relatively small number of labels, e.g., by using various combinatorial approaches.

The invention develops a double-line sequential targeted nano-drug delivery system, epigenetic regulation and traditional chemotherapy are combined, and the system is used for accurately and effectively treating acute myelogenous leukemia. The METTL3 inhibitor and the ROS-responsive daunorubicin prodrug are co-delivered in a single nano-drug, and the nano-preparation can realize continuous and double-path drug release according to the oxidation-reduction state in AML cells. In leukemia stem cells with low ROS level, STM2457 is released preferentially to induce differentiation and improve the ROS level of mitochondria, so that DA-ROS is activated, and the effect of delaying cytotoxicity is achieved. On the contrary, the AML mother cells with high ROS level can immediately trigger DA-ROS activation and release the two drugs at the same time, so that the tumor cells are directly killed. This ROS adaptive delivery strategy ensures spatial and temporal accuracy of drug release in heterogeneous AML cell populations.

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.

The invention discloses a genetically engineered bacterium for biosynthesis of gastrodin as well as a construction method and application of the genetically engineered bacterium. According to the genetically engineered bacterium, through overexpression of sucrose permease CscB derived from escherichia coli W, sucrose phosphorylase Basp of bifidobacterium adolescentis, uridine triphosphoryl-glucose-1-phosphate uridine acyl transferase UgpA of bifidobacterium bifidum and UDP-glycosyl transferase RsUGT of hedyotis diffusa, an exogenous path for synthesizing UDP-glucose from sucrose is constructed in a cell, and the UDP-glucose is synthesized into UDP-glucose. And the glycosylation capability of the gastrodin precursor on hydroxybenzyl alcohol is enhanced. The invention also provides a construction method of the genetically engineered bacterium and a method for synthesizing gastrodin through whole-cell catalysis by using the strain and taking p-hydroxybenzyl alcohol and cane sugar as substrates. The genetically engineered bacterium is high in gastrodin synthesis efficiency, the molar conversion rate can reach 97.8%, the highest yield reaches 3.61 g / L, and a new green manufacturing scheme is provided for solving the problems that a traditional gastrodin production method is low in efficiency, high in cost, large in pollution and the like.

Therapeutic compounds for red blood cell-mediated delivery of an active pharmaceutical ingredient to a target cell are described. The therapeutic compounds are configured to bind CD47 on the surface of a red blood cell and to be subsequently transferred to CD47 on the surface of the target cell, the therapeutic compound ultimately being internalized by the target cell via endocytosis. The target cell may be a cancer cell.

The invention provides application of tea catechin in preparation of a medicine for preventing and treating bortezomib-induced peripheral neuropathy. A plurality of in-vitro model results show that tea catechin can be used as a copper death inhibitor, remarkably improves related indexes of bortezomib-induced peripheral neuropathy, improves survival rates of Schwanton cells and SH-SY5Y, improves mitochondrial functions and cell cycle abnormalities of the Schwanton cells, reduces intracellular copper ion, cuprous ion and ATP levels, and can be used for preparing the copper death inhibitor for the peripheral neuropathy of the Bortezomib-induced peripheral neuropathy of the Bortezomib-induced peripheral neuropathy. Meanwhile, the death of Schwann cells and SH-SY5Y induced by the copper death inducer is reduced. In-vivo experiments show that the tea catechin plays a role in protecting functional damage of the bortezomib, and the tea catechin remarkably improves mechanical hyperalgesia, gait abnormality and pathological changes of myelin sheath and axon of mice caused by peripheral neuropathy induced by the bortezomib. The tea catechin can be used as a medicine for peripheral neuropathy induced by bortezomib, and has high clinical application value and development prospect.

The invention discloses a compound microorganism freeze-drying protective agent, a preparation method of the compound microorganism freeze-drying protective agent and a use method of the compound microorganism freeze-drying protective agent. Comprising the following raw materials in parts by weight: 0.03 part of disodium ethylene diamine tetraacetate, 5 to 15 parts of trehalose, 2 to 5 parts of sorbitol, 0.5 to 2 parts of ascorbic acid, 0.05 to 0.2 part of N-acetyl-L-cysteine, 0.1 to 0.5 part of polylactic acid-glycolic acid copolymer, 0.01 to 0.05 part of alpha-tocopherol and 1 to 3 parts of bentonite. The PLGA, the alpha-tocopherol and the NAC construct a three-stage synergistic anti-oxidation system from outside to inside: the PLGA is externally blocked, so that rapid consumption of an antioxidant is avoided, and slow release is realized; the alpha-tocopherol realizes oxidative damage repair of cell membranes; nAC maintains the internal reduction environment of cells, reduces oxidative stress and can regenerate alpha-tocopherol at the same time, and reutilization of resources is achieved.

The invention provides a DNA library screening method and application of an intracellular mRNA sensor switch. The DNA library screening method comprises the following steps: designing a DNA library containing a switch sequence of a degenerate basic group; preparing a circular expression vector library; preparing a minimum linear expression vector library; performing seamless cloning; carrying out RCA rolling circle amplification; carrying out telomerase digestion; carrying out cell transfection; extracting RNA (Ribonucleic Acid) and carrying out reverse transcription; carrying out library building of next-generation sequencing by taking the cDNA as a template; carrying out next-generation sequencing; analyzing data and comparing to obtain the DNA library of the mRNA sensor switch. According to the DNA library screening method provided by the invention, more sensor switch sequences can be provided for a single site on mRNA in cells, and more selection possibilities are further provided for subsequent targeted therapy.