89 results about "In vitro study" patented technology

The invention discloses sgRNAs of a specific targeting grass carp RIOK3 gene and a construction method and application of an RIOK3 gene knockout cell line, and belongs to the technical field of gene engineering. The invention provides sgRNAs of a specific targeting grass carp RIOK3 gene and a sequence of a primer group, and also provides a method for constructing a large-fragment knockout cell line of the RIOK3 gene in combination with the sgRNAs, and application of the gene knockout cell line in research on an anti-virus infection mechanism of grass carp. The invention discloses the construction of the grass carp gene in-vitro research model by knocking out the antiviral negative regulatory factor RIOK3 of the fish cells by using the CRISPR-Cas9 technology for the first time, provides an in-vitro research material and a new way for the analysis of an antiviral immune regulation molecular mechanism of the grass carp, and particularly has important guiding significance for the research of resisting grass carp reovirus infection.

The invention relates to the technical field of insulin resistance, in particular to an insulin resistance method for promoting a signal channel based on CTRP9. The method comprises the following steps: constructing a human hepatocyte line plasmid; carrying out cell culture and transfection; detecting the influence of CTRP9 on LO human hepatocyte glucose uptake, PGC-1a gene expression and the phosphorylation level of insulin signal channel related protein; detecting the influence of CTRP9 on glucose uptake of an LO human hepatocyte insulin resistance model, PGC-1a gene expression and the phosphorylation level of insulin signal channel related protein; and carrying out SirT1 / PGC-1a protein expression detection. According to the insulin resistance method for promoting the signal channel based on the CTRP9, a human hepatocyte line is taken as an in-vitro research object, an in-vivo research environment is simulated, a corresponding insulin resistance model is established, and the insulin resistance can be obtained by using technologies such as flow cytometry analysis, RT-PCR (Reverse Transcription-Polymerase Chain Reaction), western blotting and signal channel inhibition. And the treatment effect of CTRP9 on hepatic cell insulin resistance and the specific action mechanism are researched.

This invention belongs to the field of intestinal microbiome technology, specifically relating to a capsular polysaccharide derived from Bacteroides salorlis CSP6, its preparation method, and its applications. The monosaccharide composition of the capsular polysaccharide is mainly composed of fucose, glucose, and glucosamine, with a molecular weight of 7.139 × 10⁻⁶. 5 The strain described is deposited at the China Center for Type Culture Collection (CCTCC), accession number CCTCC NO: M20221993. In vitro studies have found that the capsular polysaccharide derived from Bacteroides Salieris CSP6 can inhibit α-glucosidase; in vivo animal experiments have demonstrated that this capsular polysaccharide can inhibit the rise in postprandial blood glucose and has a hypoglycemic effect.

Provided are methods and compositions for enhancing radiotherapy in various cancers by administering dopamine receptor (DRD2)‐binding phenylpiperazine derivatives such as brexpiprazole, cariprazine, pipamperone, and perospirone. In vitro studies show that combining these agents with ionizing radiation (e.g., 5 Gy) significantly reduces cancer cell survival, suppresses metastatic and stemness markers (e.g., CD44, MMP‑2, Snail, Nanog), and promotes apoptosis. Fractionated or single‐fraction radiation regimens can be paired with DRD2 antagonists to lower treatment‐resistant phenotypes, decrease the likelihood of recurrence, and potentially reduce necessary radiation dosages. The approach applies to breast, prostate, lung, pancreatic, and brain cancers, among others. Depending on tumor characteristics, additional chemotherapeutic or immunotherapeutic agents may further improve outcomes.

This invention relates to the field of organoid disease models, and discloses a method for constructing a human brain organoid model of early-onset Alzheimer's disease (AD) and its applications. This invention introduces [a specific technology] into human embryonic stem cell lines through single-base editing and lead editing techniques. PSEN1 ΔE9、 PSEN1 M146V and APP Human embryonic stem cell lines carrying four familial pathogenic gene mutations (K670N and M671L) were constructed and induced to differentiate into Alzheimer's disease (AD) brain organoids. The AD brain organoid model established by this invention exhibited tau phosphorylation pathological phenotypes as early as 20 days and Aβ-related phenotypes as early as 40 days, with increased total Aβ, decreased Aβ42 / Aβ40 ratio, and simultaneous aggravation of Aβ-Tau pathology. This represents a complex neurodegenerative pathological model where multiple mutations synergistically regulate Aβ production and tau phosphorylation, which is highly valuable for understanding and studying early-onset familial AD. This invention provides a human brain organoid model for in vitro AD studies and offers a tool for studying the pathological mechanisms of AD and screening drugs.

A microfluidic chip for in vitro culture, comprising: a chamber, the chamber comprising a primary channel and at least one side channel adjacent thereto, the chamber having a bottom surface and a top surface, the chamber being configured as an open-top chamber having at least one aperture extending through the top surface and in fluid communication with the primary channel; and a separator disposed between the primary channel and the at least one side channel, the separator extending from the bottom surface of the chamber toward the top surface of the chamber; and optionally, an insert, the insert having at least one projection configured obstruct access to the aperture when the projection is placed into register with the one aperture. A method, comprising: contacting a solid tumor cell residing in the primary channel of a microfluidic chip and a sample cell perfused to the primary channel.

The application discloses a human vagina micro-ecological in-vitro model and a construction method and application thereof, and relates to the technical field of medicines.The application discloses a human vagina micro-ecological in-vitro model and a construction method and application thereof.Firstly, a mixed cell group of the vaginal epithelium containing a complete cell lineage and capable of long-term subculture is successfully established, a technical bottleneck that primary vaginal epithelial cells are difficult to expand is solved, and ideal seed cells are provided for model construction.Secondly, a highly-bionic vaginal mucosa model having a tight connection structure, rich layers and an orderly arrangement is constructed, and the physiological barrier and immune regulation functions of the vaginal mucosa are truly simulated.Finally, by introducing lactobacillus symbiosis and confirming the response to estrogen, a multi-element in-vitro research model integrating the anatomical structure of the vagina, resident flora, local immunity and endocrine regulation is constructed, and a reliable technical platform is provided for in-depth exploration of the mechanism of vaginal micro-ecological disorder and efficient screening of related drugs.

This invention belongs to the medical field and provides a human trophoblastic stem cell line derived from complete hydatidiform mole and its applications. The human trophoblastic stem cell line, with accession number CGMCC No. C2024411, can be induced to differentiate into syncytiotrophoblast and extravillous trophoblast, mimicking the differentiation and developmental defects of the trophoblastic lineage in hydatidiform mole. It can be used as an in vitro research model for complete hydatidiform mole and has broad application prospects in drug target research and genetic studies.

The invention relates to a preparation method and antibacterial application of an adjustable manganese-based Prussian blue analogue. Through metal ion doping and lattice engineering regulation and control, a manganese element is introduced into a Prussian blue frame structure, and controllable regulation of the doping amount and optimization of a crystal structure are realized. The prepared material has excellent photothermal conversion capability and multienzyme simulation activity, and shows a photothermal-enzyme synergistic enhancement effect under the irradiation of near-infrared light. In-vitro studies show that the material can efficiently inhibit the growth of gram-negative bacteria; animal experiments further verify that the composition has a remarkable curative effect in infected wound repair. The invention not only provides a preparation method of a nano material with controllable metal doping, but also provides a novel photo-enzyme synergistic treatment method for drug-resistant bacterium infection treatment.

The invention relates to an application of a congenital non-ocular / microphthalmia related protein MAB21L1. The deletion and mutation of the MAB21L1 gene can cause non-ocular / microocular deformity. Specifically, the R51 site of the MAB21L1R51W mutant protein cannot form a salt bridge network with E49 and E115 sites, the protein stability is reduced, the MAB21L1R51W mutant protein is easily degraded by ubiquitin-proteasome, the expression of a key regulatory factor PAX6 for eye development cannot be promoted, and finally, the anophthalmia / microphthalmia is caused. An MAB21L1R51W human mutant cell strain and a zebra fish model are constructed based on a CRISPR / Cas9 technology, so that in-vivo and in-vitro combined research on an action mechanism of specific gene point mutation in eye development and diseases becomes possible, and a regulation mechanism of eye development diseases can be better understood; a new screening target is provided for antenatal molecular diagnosis of congenital azoopia / ommatidium diseases, and the method has positive practical significance.

The invention discloses a method for detecting the conformational change of an APO-Cascade protein. Through mutation of Cas8 protein amino acid N376 and Cas5 protein K169 in a Casade protein, the obtained Cas8 and Cas5 labeled by non-natural amino acids can maintain functions, and at the same time, the mutant type of the Cas5 / 8 protein can be subjected to fixed-point fluorescence labeling; according to the method, the Cas6 is used as a template, the N end of the Cas6 is further modified with biotin, and a formed Casade compound can be fixed on a biotin-PEG functionalized single-molecule slide, so that the conformation state of the Casade protein can be truly reflected by using single-molecule fluorescence resonance energy transfer, an effective means is provided for in-vitro research on the structure-function relationship of a system in CRISPR / Casade, and the method has a wide application prospect. And the method is of great significance in designing the Cascade protein with optimized properties and enabling the Cascade protein to play new functions.

The application provides a trophoblast organ-on-chip system for simulating a maternal-fetal interface of a systemic lupus erythematosus model based on an organ chip and a construction method thereof, the system taking an organ chip as a carrier and comprising three core parts of a trophoblast organ, an endometrial chip and an SLE disease model. The application completes separation culture and characterization of the trophoblast organ, multi-cell co-culture construction and characterization of the endometrial chip, and then constructs a pathological model, and the three parts cooperatively form a three-dimensional maternal-fetal interface. The system can clearly observe the dynamic process of the trophoblast organ invading the endometrial chip, can detect the continuous secretion of hCG and SLE-related inflammatory factors, and breaks through the defects of the prior art model, such as low authenticity, inability to dynamically observe, single detection dimension and lack of standardization, and has both physiological and pathological adhesion and experimental repeatability, can multi-dimensionally characterize the cell phenotype and function of the maternal-fetal interface, and provides an in-vitro research platform for the pathological mechanism research of the SLE pregnancy-related abnormal maternal-fetal interface.

The utility model relates to a simulation circulation device for in-vitro research of aortic diseases, which comprises a disease simulation box, a ventricular cavity, a venous cavity and an arterial cavity which are sequentially connected through a circulation pipeline to form a closed loop for simulating blood circulation flow. A first one-way valve which is in one-way conduction from the simulation box to the ventricular cavity is arranged on the circulating pipeline between the disease simulation box and the ventricular cavity; and a second one-way valve which is in one-way conduction from the ventricular cavity to the venous cavity is arranged on the circulating pipeline between the ventricular cavity and the venous cavity. The simulation circulation device for in-vitro research on the aortic diseases is low in research cost, small in risk, high in use flexibility and capable of realizing in-vitro research on the influence of the aortic diseases on cardiovascular physical parameters of a human body.

The present invention relates to the technical field of organoid culture, and for the first time relates to a culture system and a culture method for gastrointestinal tumor organoids. The culture system is composed of adding antioxidant active traditional Chinese medicine extracts such as naringin or polydatin to the existing culture system. Using this culture system to culture gastrointestinal tumor organoids can improve the primary survival rate and the proliferation rate after passage of the gastrointestinal tumor organoids. In addition, the present invention also provides an application of the culture system for gastrointestinal tumor organoids, using the culture system and the gastrointestinal cancer organoids cultured thereby to perform drug screening on small molecule targeted anti-tumor drugs, providing a better in vitro research model for subsequent anti-tumor treatment.

The invention discloses construction and application of two mouse esophageal squamous cell carcinoma organoid models with sex difference. The method comprises the steps of 1, tumor sample collection; step 2, carrying out single-cell enzymolysis treatment; step 3, modeling and culturing the tumor organoid; step 4, passage of the tumor organs and construction of a library for cryopreservation; compared with the prior art, the invention has the advantages that two stable and high-purity esophageal squamous carcinoma organoid strains are successfully established and are respectively named as PDO306 (from female mice) and PDO307 (from male mice). The establishment of the two new esophageal squamous cell carcinoma organoid strains makes up for the deficiency of immune microenvironment in the esophageal squamous cell carcinoma in-vitro research process, and valuable experimental materials and tools are provided for subsequent esophageal squamous cell carcinoma research.

The invention belongs to the field of medicine, and provides a complete grape embryo-derived human trophoblast stem cell line and application thereof. The preservation number of the human trophoblast stem cell line is CGMCC No.C2024411, and the human trophoblast stem cell line can be induced and differentiated into a syncytial trophoblast and an extravillus trophoblast, can simulate differentiation and development defects of a grape embryo trophoblast pedigree, can be applied as an in-vitro research model of a complete grape embryo, and has a wide application prospect in drug target research and genetic research.

The present disclosure provides a proteolysis-targeting compound TPB-L-E3B, a method for synthesizing the same, and use thereof. The compound can treat human tumor diseases through the eRF3a-targeting proteolysis mechanism, and exhibits great potential in treating such diseases in in-vitro studies, particularly, in treating diseases such as prostate cancer, ovarian cancer, liver cancer, cervical cancer, leukemia, breast cancer, and the like.

Aspergillus flavus is an opportunistic, saprophytic fungus that infects maize and other fatty acid-rich food and feed crops and produces toxic and carcinogenic secondary metabolites known as aflatoxins. In vitro studies showed a five-fold increase in antifungal activity of AGM182 (vs. tachyplesin1) against A. flavus. Transgenic maize plants expressing AGM182 under maize Ubiquitin-1 promoter were produced through Agrobacterium-mediated transformation. PCR products confirmed integration of the AGM182 gene, while RT-PCR of maize RNA confirmed the presence of AGM182 transcripts. Maize kernel screening assay using a highly aflatoxigenic A. flavus strain (AF70) showed up to 72% reduction in fungal growth in the transgenic AGM182 seeds compared to isogenic negative control seeds.

The invention provides a mebendazole HP-beta-cyclodextrin clathrate compound. The mebendazole HP-beta-cyclodextrin clathrate compound is prepared from mebendazole and HP-beta-cyclodextrin, wherein the mebendazole HP-beta-cyclodextrin clathrate compound is prepared from the mebendazole and the HP-beta-cyclodextrin; wherein the mebendazole is positioned in a cavity of the HP-beta-cyclodextrin. According to the mebendazole HP-beta-cyclodextrin inclusion compound provided by the invention, the mebendazole and the HP-beta-cyclodextrin are prepared into the inclusion compound, so that the water solubility of the mebendazole is improved by 18333 times and reaches 6.05 mg / mL, which is the best result obtained so far. In addition, the in-vitro PK research shows that the inclusion compound releases 80% of mebendazole within 5 min, while the pure drug only releases 20% under the same condition; in the in-vivo PK research, the bioavailability is almost doubled, which indicates that the prepared toluene imidazole HP-beta-cyclodextrin inclusion compound can be clinically applied as a potential anti-cancer drug.

The invention discloses a combined product based on NanoBiT luciferase three fragments, a method and application. The combination comprises a first fusion protein, a second fusion protein, and a third fusion protein. According to the combined product, NanoBiT luciferase is divided into three fragments, the three fragments are fused with different protein fragments, it is ensured that all the fragments of NanoBiT luciferase are integrated into a protein interaction network, therefore, non-specific signals are reduced, the biological authenticity and reliability of experimental results are improved, and compared with a traditional NanoBiT luciferase two-fragment system, the combined product has the advantages that the combined product is simple in structure and convenient to use, and the application range is wide. The light-emitting signal of protein interaction is obviously enhanced, and a more accurate tool is provided for in-vitro research of protein interaction.

The invention belongs to the technical field of biology, and particularly relates to a lymphoma organoid culture medium and a culture method. The culture medium comprises the following components: 0.5 to 2 X of a GlutaMAX additive, 0.2 to 2 X of B-27, 0.5 to 2 X of N-2, 0.8 to 1.5 X of ITS-A, 1 to 10 mmol / L of sodium pyruvate, 50 to 300 ng / ml of WNT-3A, 2.5 to 10 mmol / L of nicotinamide, and the like which are dissolved in an RPMI 1640 culture medium. According to the culture method provided by the invention, in-vitro culture can be specifically carried out on the human lymphoma tissue, and in the culture process, an organoid which is similar to an in-vivo tumor from cell constitution and space structure is formed; the culture medium provided by the invention has clear components, is suitable for culturing different types of lymphoma organoid, and can provide a favorable culture method and research model for an in-vitro research platform in the disease field.

The invention discloses a carbon tetrachloride damage model construction and nutrition intervention evaluation method based on a three-dimensional liver chip, and belongs to the technical field of microfluidics and biological medicines. According to the method, an in-vitro bionic liver tissue model is constructed by utilizing a three-dimensional liver chip, liver cell injury is induced by perfusing carbon tetrachloride, on the basis, substances such as algae-derived extracellular vesicles are added for repairing intervention, and meanwhile, contrastive analysis is performed with a two-dimensional plate culture system. Results show that the secretion level and the function maintenance time of albumin, urea and the like of liver cells in a liver chip model are superior to those of two-dimensional culture, and after nutrient intervention is carried out, liver injury indexes such as ALT, AST and the like in the chip model are remarkably reduced, and the cell activity and the metabolic function are obviously recovered. The model can be used for evaluating the curative effect of nutrient substances on chemical liver injury, overcomes animal experiment species difference and two-dimensional culture limitation, provides an efficient and sensitive in-vitro research platform for medicine and nutrition function screening, and has wide application prospects and industrialization potential.

The invention relates to the technical field of biomedical imaging, in particular to a multi-photon microscopic device and a multi-photon microscopic system, and aims at solving the problems that an existing multi-photon microscopic device is complex in structure and large in size. The multiphoton microscopic device comprises a transmission optical fiber, a scanner and an objective lens, the transmission optical fiber can realize two-way transmission of an excitation light signal and a fluorescence signal, and the fluorescence signal emitted after a target tissue is excited by the excitation light signal can be emitted into a second end of the transmission optical fiber along the objective lens and the scanner and is also transmitted by the transmission optical fiber transmitting the excitation light signal. The volume of the multi-photon microscopic device is reduced, so that the multi-photon microscopic device can be widely applied in a clinical environment, especially in a scene requiring high miniaturization, portability and in-vivo observation of the epithelium of the inner cavity of a narrow space; and a tool is provided for unmarked optical micropathology from in-vitro research to in-vivo clinical examination in the field of endothelial lesions.

The invention belongs to the technical field of biology, and discloses a reagent combination or kit for constructing an embryoid and application of the reagent combination or kit. The reagent combination or the kit can be used for inducing stem cells to generate blastocyst-like pedigree precursor cells, blastocyst-like cells and / or embryoid-like cells, is based on small molecule induction, does not depend on transgenosis, and is single in cell source, and the generated blastocyst-like pedigree precursor cells and blastocyst-like cells have a whole blastocyst pedigree, and can be used for inducing stem cells to generate blastocyst-like pedigree precursor cells and blastocyst-like cells. The seed cells are balanced in developmental ability and have totipotency, and the problem of unbalanced cell maturation generated in the prior art is solved; the generated embryoid is highly similar to a natural embryo in morphological characteristics and transcriptome characteristics, has the capability of highly reproducing embryonic development and / or protointestinal movement after implantation, and is beneficial to in-vitro research of embryonic development; the reagent combination or the kit is used for inducing stem cells to generate blastocyst-like pedigree precursor cells, blastocyst-like cells and / or embryoid-like cells, and the method is simple, high in applicability and high in embryoid-like construction efficiency.

The invention discloses a method for researching the regulation effect of extraembryonic tissues on protointestinal movement, and belongs to the technical field of biology. According to the invention, a human embryonic ectomesoderm stem cell culture system which can be stably obtained is firstly established, and the problem that related cells are difficult to stably maintain for a long time is solved. Furthermore, the human embryonic stem cells and various types of extraembryonic cells, including extraembryonic mesoderm stem cells, trophoblast stem cells and amnion-like cells, are co-cultured to construct an in-vitro simulation system for analyzing the influence of extraembryonic tissues on the occurrence of protointestinal motility. Through the method, the specific regulation effect of different types of extraembryonic cells in the formation of the protointestinal motility is disclosed, the deficiency of function cognition of extraembryonic tissues in the existing research is made up, and an efficient and controllable in-vitro research platform is provided for analyzing the early-stage protointestinal formation mechanism of human beings.

The invention discloses a flavonoid composition for targeted regulation and control of TSLP-TSLPR and application of the flavonoid composition, the flavonoid composition is prepared from kaempferol, kaempferol-7-O-glucoside and kaempferol-7-O-rhamnoside, and the molar ratio of the kaempferol to the kaempferol-7-O-rhamnoside is (1-3): (1-3): (1-3). According to the composition, generation of key cytokines IL-4 and IL-13 of allergic inflammation is remarkably inhibited by blocking a TSLP-TSLPR signal channel, so that allergic skin diseases such as TSLP-mediated atopic dermatitis, urticaria and psoriasis are effectively treated. In action mechanism research, in-vitro experiments show that when the three components are combined according to the ratio of 2: 3: 1, the strongest inhibition effect on TSLP-induced IL-4 and IL-13 secretion in a HuT78 cell model is achieved, and the effect is remarkably superior to that of a single-component treatment group. According to the invention, the action limitation of a single flavone component is broken through, the efficient targeted regulation and control of the TSLP-TSLPR pathway is realized through the synergistic effect of multiple components with accurate proportioning, and a safe and effective treatment scheme is provided for allergic skin diseases.

Disclosed are living, three-dimensional tissue constructs for in vitro scientific and medical research, arrays thereof, and methods of making said tissues and arrays.

The invention provides chloroplast storage liquid and application thereof. The chloroplast storage solution disclosed by the invention is prepared from the following components in parts by mass: 5 to 30 parts of aloe polysaccharide ABPA1, 5 to 30 parts of aloe acetylmannan ABPA2, 2 to 15 parts of red algae colloid extract and 25 to 88 parts of water. The invention also provides an application method of the chloroplast storage liquid. According to the chloroplast storage solution, the plant polysaccharide and the red algae colloid extract are applied to an in-vitro chloroplast storage system, so that the activity of chloroplast can be effectively maintained for a long time, and a stable and reliable in-vitro research window is provided for deeply analyzing a fine mechanism of photosynthesis; moreover, it is possible to serve as a high-efficiency biological light energy converter.

The invention belongs to the fields of veterinary medicine and medical biology, and particularly relates to canine origination and application of a canine IL-31 monoclonal antibody. Aiming at the monoclonal antibody of the targeted canine IL-31, a mouse monoclonal antibody of the targeted canine IL-31 is produced by a mouse hybridoma technology, and then the canine source antibody of the targeted canine IL-31 is produced by modifying an FW region of the antibody. In-vitro researches prove that the antibodies have activity in vitro and can be applied and produced as a therapeutic scheme for inhibiting canine allergic pruritus in the future.

The invention relates to the field of biomedicine, in particular to a mouse oligodendroglia precursor cell culture method which mainly comprises the following steps: taking cerebral cortex tissues of a 3-day-old mouse, cutting the cerebral cortex tissues into pieces, digesting the cerebral cortex tissues to obtain mixed glial cells, and inoculating and culturing the mixed glial cells; and after culturing for 10 days, carrying out mechanical oscillation on a shaking table (the rotating speed is 250rpm, and the time is 6 hours), and separating to obtain the oligodendrocyte precursor cells with high purity and high activity. The method is easy and convenient to operate and high in repeatability, a large number of mouse oligodendroglia precursor cells can be obtained only through a conventional, simple and cheap culture medium, and the cells have the continuous multiplication capacity, can be continuously passaged and have the capacity of being differentiated into mature oligodendroglia cells; the method can be widely applied to in-vitro research of special physiological functions and related biological mechanisms of the mouse oligodendrocyte precursor cells.