39 results about "Vacuole" patented technology

The invention discloses a single vesicle particle electrochemical signal identification and classification method and identification system, and the method comprises the steps: S1, preparing a carbon fiber ultramicro electrode, applying a voltage in a patch clamp system constant potential mode, and collecting picoampere-level current response generated by oxidation of a single vesicle content on the surface of the electrode; s2, carrying out smooth filtering on the collected original current signal, and realizing characteristic peak identification and multi-dimensional physical / statistical characteristic extraction by using a multi-parameter peak identification method based on a threshold value; s3, carrying out preprocessing operations including missing value processing, standardization, principal component analysis dimension reduction and category equalization on the extracted peak value characteristic data; and S4, on the basis of the preprocessed feature data, constructing and training at least one machine learning model for automatically identifying the particle size category of the vesicle particles. The method combines the advantages of single-vesicle electrochemical high-sensitivity measurement and intelligent algorithm classification, and has the advantages of high recognition efficiency, good repeatability, wide applicability and the like.

A genetically modified yeast cell, wherein the yeast cell comprises a genetic modification comprising overexpression of yeast gene encoding porphobilinogen deaminase (HEM3), the HEM3 gene having at least 80% identity with SEQ ID No. 7. The genome of the modified yeast cell further comprises one or more genetic modifications in one or more genes selected from: genes coding for heme-dependent repressor of hypoxic genes (ROX1), genes coding for heme oxygenase (HMX1), genes coding for a receptor for vacuolar proteases (VPS10), and genes coding for vacuolar proteinase (PEP4), the one or more genetic modifications being such that expression of a polypeptide from such a gene is reduced or disrupted or the polypeptide expressed is non-functional.

The invention discloses a method for preventing and controlling cadmium accumulation of rice grains and strengthening zinc concentration at the same time, and relates to the technical field of agricultural environment and crop nutrition regulation, the single regulation defect is precisely overcome through the synergistic effect of a compound foliage regulator, salicylic acid and a zinc amino acid chelate are compounded to form the regulator, and the regulation effect is improved. Salicylic acid is utilized to induce synthesis of plant chelating peptide in rice, cadmium vacuole partitioning is enhanced, cadmium transporter protein expression is reduced, and migration of cadmium to grains is prevented and controlled from a transport path; meanwhile, zinc is rapidly supplemented by means of the efficient absorption characteristic of the zinc amino acid chelate, and the zinc amino acid chelate can cooperate with a salicylic acid signal to further inhibit cadmium distribution, so that the problems that a single zinc fertilizer cannot resist cadmium and single salicylic acid cannot be rich in zinc are solved, cadmium resistance and zinc enrichment are synchronously promoted, and meanwhile, dry-wet alternate irrigation in the booting stage and the mature stage is matched, so that the yield is increased. By regulating the soil moisture state, the physiological metabolism environment of the rice is optimized, the absorption and utilization of a regulating agent are promoted, and the vacuole partitioning effect of cadmium and the phloem transfer efficiency of zinc are enhanced.

The invention discloses a synbiotics-like feed prepared from vegetable waste polymerized vegetable cakes and a method, and relates to the field of agricultural circular economy, cellulose in vegetable waste cell walls is effectively decomposed under mild conditions through a specific catalytic decomposition enzyme combination, and part of cellulose is converted into carbon dioxide and water, so that the utilization rate of the cellulose is improved. By combining physical heat energy conduction, liquid in cell vesicles is heated to generate micro air pressure to break through residual cell walls, so that recessive water in cells is successfully converted into surface dominant water, the energy consumption in the dehydration process is greatly reduced, and the loss of dry basis substances in the resource utilization process is effectively reduced; the vegetable cake product obtained through final polymerization is rich in various nutrient substances such as crude fibers and / or dietary fibers, proteins, vitamin C and carbohydrates, the recycling rate of the vegetable waste is larger than or equal to 95%, the recycling efficiency and the additional value are remarkably improved, meanwhile, the difficulty of subsequent harmless treatment is reduced from the source, and the economic input and the treatment cost are reduced.

The invention discloses application of TPN171H, an active fragment thereof or a derivative thereof in preparation of a medicine for treating spinal cord injury. The TPN171H shows an excellent treatment effect in spinal cord injury treatment, the levels of apoptosis factors BAX and Caspase-3 cleaved can be remarkably reduced, the level of Bcl-2 is improved, the content of a neuron marker MAP-2 is also remarkably increased, and the number of Tunel bodies and the cavitation-like structure are remarkably reduced. The intervention of the TPN171H also can reduce MNSS, and prompts that the TPN171H has the functions of promoting the spinal cord movement and coordination of the rat with spinal cord injury and improving the sensory system of the rat with spinal cord injury.

This invention relates to the field of anti-aging drug technology, specifically to the application of small molecule compounds in the preparation of anti-aging drugs. These small molecule compounds competitively block the interaction between KDM6A and the vacuolar ATPase V0 domain subunit ATP6V0C by targeting a specific small molecule binding pocket of the histone demethylase KDM6A, thereby inhibiting the demethylation activity of KDM6A on lysine residue 36 of ATP6V0C. These small molecule compounds can effectively downregulate aging-related β-galactosidase activity, improve cell cycle arrest, and reverse the senescent phenotype of nerve cells induced by the environmental pollutant GenX, making them particularly suitable for combating premature aging of nerve cells induced by perfluorinated and polyfluoroalkyl compounds such as GenX. This technical solution can solve the technical problem of the lack of drugs with clearly defined components, a clear mechanism of action, stable efficacy, and the ability to specifically alleviate aging in existing technologies, and has ideal prospects for widespread application.

The invention relates to the technical field of anti-aging drugs, in particular to application of a small molecule compound in preparation of anti-aging drugs. The small molecule compound competitively blocks the interaction between the KDM6A and a vacuolar ATP enzyme V0 structural domain subunit ATP6V0C through a specific small molecule binding pocket of targeted histone demethylase KDM6A, and further inhibits the demethylation activity of the KDM6A on 36th lysine of the ATP6V0C. The small molecule compound can effectively down-regulate the activity of senescence-related beta-galactosidase, improve cell cycle arrest and reverse nerve cell senescence phenotypes induced by environmental pollutant GenX, and is especially suitable for resisting nerve cell premature senescence caused by perfluorinated and polyfluoroalkyl compounds such as GenX. According to the technical scheme, the technical problem that drugs which are definite in component, clear in action mechanism, stable in curative effect and capable of pointedly relieving senescence are lacked in the prior art can be solved, and ideal application and popularization prospects are achieved.

The invention discloses a preparation method of a coronary artery microcirculation disturbance animal model, and belongs to the technical field of biological medicine. According to the method, the stable mouse CMD model can be established by injecting 20-40 [mu] g / mL Calcimycin into the cardiac apex for the first time. Calcimycin is a calcium ion (Ca < 2 + >) carrier commonly used in a laboratory and can transport calcium ions outside cells into the cells, so that the concentration of the calcium ions in the cells is increased. Particularly, when 40 [mu] g / mL of Calcimycin is injected, the pathological severity of the Calcimycin is further aggravated, myocardial fiber arrangement is disordered, boundary is unclear, a large number of myocardial cells are necrotic and dissolved, cavitation is formed, vascular wall degeneration swelling is caused, inflammatory cell infiltration is caused, and thrombus is formed in microvessels.

This invention discloses a genetically engineered bacterium for synthesizing carotenoids, its construction method, and its applications, belonging to the field of genetic engineering technology. The genetically engineered bacterium uses a carotenoid-producing yeast strain as the starting material. The gene ROX1, encoding a heme-dependent hypoxia gene repressor, is knocked out, while the expression of genes STB5 (encoding a transcription factor involved in NADPH regeneration), DID2 (encoding a class E protein in the vacuole protein sorting pathway), and VOA1 (encoding an endoplasmic reticulum protein playing a role in the assembly of the V0 region of V-ATPase) is upregulated. This invention promotes carotenoid synthesis and significantly increases carotenoid yield through combined regulation of gene expression outside the carotenoid synthesis pathway, demonstrating promising application prospects.

The invention relates to the technical field of microbial fermentation and genetic engineering, and particularly discloses a yeast engineering bacterium for efficiently producing plastic depolymerases and application of the yeast engineering bacterium. The engineering yeast takes pichia pastoris as an expression host, a molecular chaperone is co-expressed, a vacuole sorting receptor gene is knocked out or knocked down, in addition, a yeast cell factory for efficiently expressing the plastic depolymerases is constructed by further combining regulation and control element optimization and target gene copy number regulation, and the engineering yeast has the advantages that the engineering yeast can be used for efficiently expressing the plastic depolymerases; the regulatory element comprises any one or a combination of more of a strong promoter, a signal peptide and a C-terminal tag. The engineering yeast provided by the invention can realize efficient accumulation and secretion of the plastic depolymerases, obviously improve the enzyme yield and reduce the production cost of unit enzyme activity, and provides a stable and efficient yeast expression system for industrial and low-cost production of the plastic depolymerases; the method has an important application prospect in the fields of biodegradation and resource utilization of waste plastics.

The present application relates to lipid vacuole (aka LV)-containing lipocartilage cells (aka LCs), and methods for the differentiation of human lipocartilage progenitors into LV-containing LCs. Also disclosed are methods related to purifying, labeling, maturing, and delivering of LV-containing LCs to a subject in need thereof.

Methods and compositions are provided for enhancing the accumulation and / or recovery of recombinant casein expressed in soy. Endogenous Casein-Processing Interfering Proteins (CPIPs), such as oxidases like lipoxygenases (LOX), are present in the recombinant protein fractions and interfere with yield and / or purity. RNA interference (RNAi) mechanisms are leveraged in plasmids that knockdown genes expressing CPIPs and / or genes in the Soybean Vesicular Trafficking Pathway (SVTP). Knocking down of two or more target genes is manifested using a dual gene targeting approach, often via a single expression cassette containing multiple distinct shRNA constructs for enhanced efficiency. The shRNA constructs invoke a RNAi mechanism that leads to increased casein accumulation either: (1) directly, by suppressing CPIPs (e.g., LOX) to enable more efficient purification; or (2) indirectly, by modifying SVTP genes to route casein away from lytic vacuoles, preventing degradation.

The invention discloses staining agent uniform smearing equipment for a clinical laboratory, and belongs to the technical field of medical instruments, the staining agent uniform smearing equipment comprises an automatic smear conveying belt, the automatic smear conveying belt is provided with a plurality of smear placing mechanisms, each smear placing mechanism is provided with a bubble left-right swing removing mechanism, and the top of the automatic smear conveying belt is provided with a uniform smearing mechanism. And a bubble rolling removal mechanism is arranged on the automatic smear conveying belt. The bubble left-right swing removing mechanism works to push deep bubbles to the surface, the bubble rolling removing mechanism thoroughly discharges surface bubbles and corner-containing tiny bubbles, and for areas such as four corners and the edge of a smear where the bubble left-right swing removing mechanism cannot reach, due to the fact that stress on the edge of the smear is weak during swinging, tiny bubbles are prone to remaining, and the smear is not prone to falling off. The bubbles at the corners can be thoroughly removed through rolling coverage of the driving wheel, so that the surface of the smear is not blocked by any bubble, the situation of misjudgment of cell vacuoles cannot occur during subsequent microscope observation, and the blood cell morphology recognition accuracy is improved.

The invention discloses a biphase gel material formed based on hydrogen bond self-assembly and a preparation method and application thereof.The biphase gel material is formed by a first polymer and a second polymer based on hydrogen bond self-assembly, the first polymer comprises a branched polymer, the second polymer comprises a long-chain linear multi-hydrogen-bond site polymer, and the long-chain linear multi-hydrogen-bond site polymer comprises a long-chain linear multi-hydrogen-bond site polymer. The branched polymer comprises at least one of the following substances: 1) a polyether polymer with hydroxyl at the tail end; and 2) a polymer obtained by carrying out a derivatization reaction on the hydroxyl of the polyether polymer in the step 1). A biological inert film can be formed in situ on the surface of the water-loss gel, so that invasion of exogenous bacteria is effectively blocked. A skeleton-vacuole structure is formed inside due to microscopic phase separation, so that a large number of bacteria or cells and the like can be loaded. The unique internal and external structures of the two-phase gel material can load wide and complex contents and provide a steady state for the contents, and meanwhile, the two-phase gel material can be widely applied to skin diseases, implant materials and the like due to excellent mechanical properties of the two-phase gel material.

The application provides a compound, a preparation method and application in preparation of a medicine for treating tumors, and relates to the technical field of chemical synthesis. The compound can stably induce cells to produce pyroptosis characteristic cell membrane large bubbles, simultaneously induce intracellular formation of a large vacuole-like structure similar to the characteristics of vacuole death and paraptosis, and has significant tumor cell proliferation inhibition activity, thereby providing a new chemical entity and research idea for research and development of a new type of anti-tumor medicine and research of new cell death mechanisms such as pyroptosis, vacuole death and paraptosis.

The invention discloses a natural hirudin extraction method and application, and relates to the technical field of natural hirudin extraction.The method is characterized by comprising the steps that a tissue structure capable of being subjected to salivary gland single cell combined gland culture is selected; washing away bloodstain in the tissue structure by using PBS, then washing for three times by using the antibacterial liquid, and finally washing away the residual antibacterial liquid by using PBS; inoculating the tissue structure to a cell culture plate, placing the cell culture plate in a 28 DEG C incubator for inverted culture for 2 hours, then adding a DMEM / F-12 complete medium containing a growth additive and not containing serum, and placing the cell culture plate in the 28 DEG C incubator; when the tissue structure generates a large number of secretory vesicles, collecting the culture solution, adding a new culture solution, taking the collected culture solution as a natural hirudin crude product, and separating and purifying the natural hirudin crude product to obtain the natural hirudin. The method can achieve the effects of long-term survival in vitro and generation of a large amount of secretions.

The invention relates to recombinant pichia pastoris and application of the recombinant pichia pastoris in production of Brazilian sweet protein, and belongs to the technical field of fermentation. According to the invention, in the fermentation process of the Brazilian sweet protein, part of the Brazilian sweet protein is gathered and degraded in vacuoles, so that the Brazilian sweet protein secreted to extracellular cells is reduced, and finally, the yield of the Brazilian sweet protein in fermentation liquor is not high. Through research, the invention finds that the knockout of the protease with the amino acid sequence as shown in SEQ ID NO.2 or SEQ ID NO.3 can improve the exocytosis amount of the brazitame protein, related process parameters in the fermentation process are further optimized, the yield of the brazitame protein in a 3L fermentation tank reaches 4.01 g / L, and the high yield of the brazitame protein is realized.

The present invention provides a system of polynucleotides including: (a) a first polynucleotide including a nucleic acid sequence encoding a sorting signal sequence selected from an apoplast sorting signal sequence or a vacuole sorting signal sequence and a milk protein; (b) a second polynucleotide including a nucleic acid sequence encoding a sorting signal sequence selected from an apoplast sorting signal sequence or a vacuole sorting signal sequence and a first copy of a mammalian kinase; and (c) a third polynucleotide including a nucleic acid sequence encoding a Golgi sorting signal sequence and a second copy of said mammalian kinase, wherein the first polynucleotide, the second polynucleotide, and the third polynucleotide are operably linked. Further provided are cells and compositions including the system of polynucleotides, as well as methods of suing same, such as for producing a recombinant milk protein or a plurality thereof.

The invention belongs to the field of marine natural product chemistry, and particularly relates to a marine fungus-derived alkaloid derivative and application thereof in prevention and treatment of pepper phytophthora blight. The marine fungus crude extract disclosed by the invention shows very strong antifungal activity on phytophthora capsici under the concentration of 300 mu g / mL. Seven monomeric compounds are separated and identified, NEA shows strong antifungal activity to phytophthora capsici and is stronger than positive control azoxystrobin, the NEA effectively inhibits hypha growth, sporangium formation and zoospore movement of phytophthora capsici hyphae, and meanwhile, the protective ability of pepper plants is enhanced; nEA causes obvious morphological change of phytophthora capsici protonema, so that cell membrane integrity damage, vacuole abnormal expansion, vacuole rupture, serious plasm-wall separation and disappearance of most organelles are caused. In addition, the content of MDA in the phytophthora capsici mycelium induced by the NEA is increased, the activity of antioxidant enzyme is reduced, and the level of soluble protein is reduced.

Providing a new method for determining gene mutations in JMML. [Solution] A method for determining gene mutations in juvenile myelomonocytic leukemia (JMML), comprising the step of determining that the subject has gene mutations in the CBL group or NF1 group if the distribution of monocyte morphology in a blood sample taken from the subject is as follows: (1) if there is a lot of A and a little D, the subject is in the PTPN11_somatic group; (2) if there is a lot of B, the subject is in the PTPN11_germline group; (3) if there is a lot of C, the subject is in the KRAS group; (4) if there is little A and C and a lot of B and D, the subject is in the NRAS group; or (5) if the subject does not fall into any of the above categories (1) to (4), the subject is in the CBL group or NF1 group. A is a monocyte that has cytoplasmic vacuoles and a club-shaped nucleus; B is a monocyte that does not form a cytoplasmic vacuole and has a club-shaped nucleus; C is a monocyte that has cytoplasmic vacuoles and a round nucleus; D represents a monocyte that does not form a cytoplasmic vacuole and has a round nucleus.

The application discloses a single-vacuole particle electrochemical signal recognition classification method and a recognition system, and the method comprises the following steps: S1, a carbon fiber ultramicroelectrode is prepared, and a voltage is applied in a membrane patch clamp system constant potential mode, and a picoampere-level current response generated by oxidation of single-vacuole content on the surface of the electrode is collected; S2, the collected original current signal is subjected to smoothing filtering, and a multi-parameter peak recognition method based on a threshold value is used to realize feature peak recognition and multi-dimensional physical / statistical feature extraction; S3, the extracted peak feature data is subjected to a preprocessing operation including missing value processing, standardization, principal component analysis dimension reduction and class balancing; and S4, at least one machine learning model is constructed and trained based on the preprocessed feature data, and is used for automatically recognizing the particle size class of the vacuole particle. The application combines the advantages of single-vacuole electrochemical high-sensitivity measurement and intelligent algorithm classification, and has the advantages of high recognition efficiency, good repeatability and wide applicability.

The invention discloses an application of polygonatum cyrtonema polysaccharide (PCP1) in preparation of an anti-depression drug. The polysaccharide is extracted from rhizomes of polygonatum cyrtonema and is proved to be used for preparing antidepressant drugs and drugs for improving expression of hippocampal brain-derived neurotrophic factors (BDNF) for the first time. Experiments show that the PCP1 can significantly improve depression-like behaviors, inhibit pathological changes such as hippocampal neuronal vacuole and immobilization, effectively up-regulate BDNF protein expression and reverse abnormal biochemical indexes of related serum. The compound has the characteristic of multiple target points, and shows a good patent medicine prospect.

The present application belongs to the field of biotechnology, and specifically relates to a method for expressing human serum albumin by using Nicotiana sylvestris NS suspension cell line, which comprises the following steps: establishing a stably cultured Nicotiana sylvestris suspension cell system, and producing by amplification, and the obtained cell biomass can reach up to 500 g / L. Then, taking human serum albumin as the research object, transformation screening is carried out in the Nicotiana sylvestris NS suspension cell system to realize high expression of HSA. The present application removes the complex tissue regeneration step, and through plant codon optimization and subcellular targeting strategy design, aims to overcome the problem of low expression of exogenous genes in plants, so as to realize efficient enrichment of target proteins in specific organelles (such as vacuole / endoplasmic reticulum, etc.). The technical scheme of the present application can be popularized and applied to the expression and screening of other types of functional proteins, and provides important technical reference and practical support for the application of plant suspension cell lines in the field of recombinant protein expression.

The invention discloses a preparation method of tomato leaf vacuole-free cytoplasm. The method comprises the following steps: cutting tomato leaves, putting the cut tomato leaves into an enzymolysis buffer solution containing cellulase, macerozyme, mannitol, potassium chloride, MES, calcium chloride, beta-mercaptoethanol and fetal calf serum, and carrying out vacuumizing enzymolysis to obtain protoplast; removing vacuoles through density gradient centrifugation, and collecting tiny protoplasts; and performing staged homogenate cracking and centrifugal purification to obtain the tomato leaf vacuole-free cytoplasm. The vacuole-free cytoplasm prepared by the invention has dsRNA cleavage activity, and can be used for tomato virus in-vitro research and plant cell-free system research and development.

The invention provides recombinant saccharomycetes and application thereof in fermentation production of collagen, and relates to the technical field of bioengineering. According to the invention, it is found for the first time that any one or two of vacuole aspartic protease, vacuole protease B, carboxypeptidase Y and GPI-anchored aspartic protease in the recombinant saccharomycetes can improve the yield of collagen in the manner of inhibiting expression or not expressing the recombinant saccharomycetes. According to the application, the proteases are also subjected to combined knockout, and results show that when the vacuole aspartic protease and the vacuole protease B are knocked out at the same time, the vacuole aspartic protease and the carboxypeptidase Y are knocked out at the same time, and the vacuole aspartic protease and the GPI-anchoring aspartyl protease can effectively improve the collagen yield of the recombinant saccharomycetes. A novel engineering bacterium and a construction thought are provided for producing collagen by recombinant saccharomycetes, and the method can be widely applied to collagen industrialization.

The invention provides an application of JTC801 as a macrovesicle-like cell death inducer in tumor resistance. Specifically, the invention relates to JTC801 as a small molecule kinase inhibitor drug, the JTC801 has a good death induction effect on tumor cells, especially uveal melanoma cells, and the induced cell death form is not classical apoptosis, pyroptosis, ferroptosis and the like, but is a giant vesicle-like death mode. The death mode is similar to giant vesicle type death, a large number of vacuoles are accumulated in cytoplasm, but different from the giant vesicle type death mode, the death mode does not have the phenomenon that fusion of lysosome and macropinosome is inhibited, and therefore the death mode is defined as the giant vesicle type death mode. Therefore, the JTC801 as a newly discovered macrovesicle-like cell death inducer has a good application prospect in tumor treatment, especially in treatment of uveal melanoma.

This method for measuring transporter activity comprises: exposing a membrane vesicle having a vacuolar ATPase and a vesicular nucleotide transporter to an anion which is labeled with a radionuclide and is different from adenosine triphosphate, in a reaction solution containing adenosine triphosphate and magnesium ions; and measuring the transport activity of the anion into the membrane vesicle on the basis of the radiation of the radionuclide.

The invention discloses an ecological oxidation processing method of oolong tea, which takes the ratio of free water / bound water of tea green cells as a key index and runs through the stages of fine manipulation, fixation and drying. By manually simulating the northeast wind climate environment and combining the synergistic effect of chloroplast, vacuoles and mitochondria, aerobic respiration and tricarboxylic acid circulation of tea leaves are restarted and maintained, and accumulation and transformation of contained substances are promoted. According to the method disclosed by the invention, the quality of the tea leaves is remarkably improved, the finished tea soup is fragrant, clear, sweet and active, the flavor is harmonious, the tea soup is neutral and slightly alkaline, the tea soup has stable flower and fruit fragrance, and a brand-new theoretical view angle and a practical path are provided for tea leaf processing.

The invention discloses a method for constructing an alcoholic liver injury animal model and application, and belongs to the technical field of gene editing. The technical problem to be solved by the invention is how to construct the alcoholic liver injury animal model. The method for constructing the alcoholic liver injury animal model disclosed by the invention comprises the following steps: knocking out an animal Cfhr3 gene to obtain a Cfhr3 gene knockout animal; and performing intragastric administration on the Cfhr3 gene knockout animal with ethanol or an aqueous solution thereof to obtain the alcoholic liver injury animal model. The animal model constructed by the method shows that after ethanol gavage, liver tissue has balloon-like change, obvious fat vacuoles appear, hepatic cord structures disappear, hepatocytes are swollen, the liver tissue is damaged, the liver index is improved, and the content of glutamic-pyruvic transaminase and glucose in blood is increased. The alcoholic liver injury animal model is successfully obtained.