1701 results about "Intracellular" patented technology

Systems and methods are provided for stabilization of cells and retention of biomolecules therein for further analysis. Block copolymers are inserted into membranes of biological particles, cell membranes or organelles (e.g., organelle membranes) and polymerized to form an intra-biological particle (e.g., intracellular) polymer network.

The invention belongs to the technical field of biology, and discloses a stable protective agent, a cell antigen dry sheet and a preparation method and application of the dry sheet, the stable protective agent comprises glycerin with the volume fraction being 0.4%, gelatin with the mass volume fraction being 0.1%, EDTA with the mass volume fraction being 0.1 mM, trehalose with the mass volume fraction being 0.03% and resveratrol with the mass volume fraction being 100 [mu] M. The stable protective agent is adopted for preparing the cell antigen dry sheet, and the cell antigen dry sheet is applied to CBA detection. The stable protective agent has good permeability, can penetrate through cell membranes to enter cells, provides a protection effect inside and outside the cells, has moisture retention and oxidation resistance, can prevent cell deformation and cell fixing layer shedding, and can well maintain the stability and reliability of antigen proteins, and the prepared cell antigen dry sheet can be stored for a long time, and has a good application prospect. The antigenicity of a membrane signal index is not damaged. When a positive sample is detected, obvious green fluorescence cells can be seen, red fluorescence quenching is avoided, and co-localization of red fluorescence and green fluorescence can be observed conveniently.

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 invention discloses application of MIGA2 as a therapeutic target in a medicine for preventing and treating Alzheimer's disease, and belongs to the technical field of biological medicine. The MIGA2 gene is used as a therapeutic target to be applied to development, screening or preparation of drugs for preventing and treating Alzheimer's disease. After an MIGA2 overexpression plasmid is adopted to transfect an Alzheimer's disease cell model, it is found that autophagy flow in cells can be promoted and accumulation of related toxic proteins can be reduced, it is proved that MIGA2 has the neuroprotective effect on the Alzheimer's disease, and a new way and exploration direction are provided for treatment of the Alzheimer's disease.

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 discloses organic nano composite hydrogel as well as a preparation method and application of the organic nano composite hydrogel, and belongs to the technical field of biological medicine delivery. The polysaccharide-based nano prodrug in the composite hydrogel can be subjected to surface protonation under the stimulation of a tumor extracellular slightly acidic environment, so that cellular uptake is promoted, low-pH / high-concentration glutathione in tumor cells is responded, intracellular aggregation and controllable release of the nano drug are realized, the retention time of the drug in the cells is prolonged, and the bioavailability of the drug is improved. The technical effect of killing tumor cells through combined chemotherapy is achieved. According to the preparation method disclosed by the invention, the release of the polysaccharide-based nano prodrug from the hydrogel and the tissue infiltration capacity are accelerated by adopting fluorinated orthoester, the infiltration of the polysaccharide-based nano prodrug to cells is increased, and then the drug-loaded compound is encapsulated by using gellan gum, so that the slow-release effect of the polysaccharide-based nano prodrug is enhanced; therefore, the efficient enrichment of the medicine at the tumor part is realized.

The present invention relates to a fusion protein that produces point mutations in cells, its preparation and use. Specifically, the fusion protein provided by the present invention contains a Cas enzyme that lacks cytosine deaminase and nuclease activity and retains helicase activity, or is formed by a Cas enzyme that lacks cytosine deaminase and nuclease activity and retains helicase activity. The present invention also relates to the coding sequence of the fusion protein, a polynucleotide sequence containing the coding sequence, a nucleic acid construct containing the polynucleotide sequence, a corresponding host cell, a method for producing point mutations in a cell, and a kit, etc. By using the present invention, it is possible to achieve site-directed mutagenesis while obtaining high mutation efficiency and a variety of mutation combinations in a specific gene region.

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 provides an organic molecule cytoplasm delivery carrier as well as a preparation method and application thereof, the compound for cytoplasm delivery has the characteristic of fat-soluble aggregation, and can form electrostatic adsorption with small molecule protein goods such as RNase A (13.7 KDa), Trypsin (24KDa) and Saporin (32.8 KDa), medium molecule protein goods such as HRP (40KDa) and macromolecular protein goods such as beta-Gal (430KDa), so that the compound for cytoplasm delivery can be used for preparing a lipid-soluble protein carrier. And furthermore, transmembrane transportation is formed through the strong fat solubility property, so that the protection and intracellular delivery of carried protein molecules are realized. According to the delivery mechanism, cargo molecules can quickly penetrate through cell membranes, and carried protein molecules can enter cytoplasm to exert functions without release operation. Development of the organic molecule cytoplasm delivery carrier is expected to assist potential protein therapies with low bioavailability and off-target toxicity to complete clinical transformation, which is crucial to tumor therapy, gene therapy and the like requiring precise local administration.

The invention provides a plug-and-play Mi3 protein nanocage fusion protein with a VNP6 tag and a preparation method and application of the plug-and-play Mi3 protein nanocage fusion protein, the fusion protein comprises the VNP6 tag, a SpyCatch003 component, a Mi3 protein nanocage and a functional protein sequence, and the VNP6 tag is located at the N end of the fusion protein. The VNP6 peptide sequence or the variant thereof is utilized to induce a vesicle or vesicle-like structure beneficial to protein folding and stabilization in escherichia coli, so that the correct folding rate and activity of recombinant protein are improved while the overall intracellular molecular crowding effect is relieved. The core idea of the invention lies in that the internal structure of cells is regulated at low temperature, and the internal microenvironment of an escherichia coli expression system is optimized, so that protein molecules under high expression load can obtain more reasonable spatial distribution, and the problems of misfolding and aggregation caused by molecular crowding are reduced.

Compositions comprising multiple pHLIP® peptides linked to a lipid nanoparticle encapsulating one or more nucleic acids (pHLIP®-LNP) and methods of preparing the pHLIP®-LNPs are described.

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.

Disclosed herein are a peptoid compound and a peptoid conjugate such as a peptoid-lipid conjugate, and preparation methods therefor and the use thereof in a small molecule drug and nucleic acid delivery. Further disclosed herein are a lipid particle containing the peptoid-lipid conjugate, such as a liposome and a lipid nanoparticle, and a small molecule drug and / or a nucleic acid delivery composition containing the peptoid-lipid conjugate or the lipid particle. The peptoid-lipid conjugate, lipid particle, and small molecule drug and / or nucleic acid delivery composition that can be used for the small molecule drug and nucleic acid delivery of the present invention enable highly efficient compounding, protection, intracellular and targeted delivery and release of the small molecule drug and biomolecules, such as nucleic acids, in tissues and organs both in vitro and in vivo.

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 relates to the technical field of biology, in particular to a tumor postoperative vaccine as well as a preparation method and application thereof. The method comprises the following steps: resuspending tumor cells in a first solution containing metal ions for incubation, collecting the tumor cells, resuspending the tumor cells in a second solution containing mineralized ions for incubation, enabling the metal ions to react with the mineralized ions to obtain metal inorganic salt, forming a mineralized shell layer on the surfaces of the tumor cells, and resuspending the tumor cells to obtain the tumor postoperative vaccine. And due to the mineralized shell layer, the vaccine has the capability of activating a signal channel in an antigen presenting cell through mechanical stimulation, so that an immune system is favorably activated, and the anti-tumor capability of the vaccine is improved. In addition, tumor cells are inactivated in the vaccine preparation process and keep a complete form, and presentation of a complete tumor antigen is facilitated, so that a relatively good anti-tumor effect is obtained.

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 present invention relates to a hybrid exosome formed by the fusion of a human cell-derived exosome and an artificial exosome. When an active ingredient labeled with a fluorescent material was attached or encapsulated in the hybrid exosome of the present invention and applied to cells, the active ingredient was observed in the cells. In addition, when the exosome was applied to skin tissue, fluorescence expression could also be observed in the dermal layer. Therefore, the hybrid exosome according to the present invention can be used as a novel drug delivery system.

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.

Disclosed are fusion proteins (and homodimers and heterodimers thereof). Fusion proteins can bind to a protein of interest and to an internalizing receptor (transferrin receptor) on a cell surface. Once bound, the protein of interest can be internalized and / or degraded inside a cell.

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 disclosure relates to the synthesis of cyclic peptides that are able to effectively deliver cargo, e.g., a therapeutic moiety (TM), inside a cell to treat a variety of conditions and diseases.

The invention belongs to the technical field of biological medicine, and discloses a preparation method of ionizable cationic lipid. The preparation method comprises the following steps: reacting 7-oxoheptyl caprate with hydroxylamine or hydroxylamine salt in the presence of alkali in the presence of a reducing agent and a cobalt-ruthenium alloy catalyst; (9Z, 12Z)-octadecane-9, 12-dienal is added into the reaction product, the reaction continues in the presence of a reducing agent, and a hydroxylamine derivative intermediate is obtained; and performing amidation reaction with 4-dimethylaminobutyric acid or hydrochloride thereof in the presence of a condensing agent and a catalyst to obtain the ionizable cationic lipid. The invention also discloses an application of the ionizable cationic lipid, the lipid nanoparticle composition or the mRNA-loaded lipid nanoparticles obtained by the preparation method in preparation of a pharmaceutical composition or a kit for mRNA delivery. The LNP provided by the invention has good physicochemical properties, biocompatibility and efficient mRNA encapsulation and intracellular delivery capabilities, and can be applied to preparation of pharmaceutical compositions or kits for mRNA delivery.

The invention relates to the technical field of biology, in particular to a human cholangiocarcinoma drug-resistant cell line HuCCT1-GEM-2 and application thereof.The human cholangiocarcinoma drug-resistant cell line HuCCT1-GEM-2 is preserved in the China Center for Type Culture Collection on May 11, 2023, the preservation number is CCTCC NO: C2023103, the IC50 value of the human cholangiocarcinoma drug-resistant cell line HuCCT1-GEM-2 to gemcitabine is 314.5 mu M, the IC50 value of a parent cell HUCCT1 to gemcitabine is 12.7 mu M, the drug-resistant index is up to 24.8, and the human cholangiocarcinoma drug-resistant cell line HuCCT1-GEM-2 can be used as a drug-resistant cell line. The cell body is obviously enlarged and rounded or is more irregular in shape, the specific volume of the cell cytoplasm is larger, binuclear or multinuclear appears in the cell, and the drug-resistant cell presents obvious bulk aggregation growth; after 24 hours, the growth rate of the HuCCT-1-GEM-2 cell is obviously slower than that of a parent cell, and the HuCCT-1-GEM-2 cell can be used for researching the drug resistance mechanism of bile duct 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.