56 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.

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.

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 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 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.

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 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 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 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.

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 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.

Disclosed herein are fully synthetic polymer-based lung surfactant materials, for the first time, as next generation SRT. In vitro studies on these polymer lung surfactants show that the candidate materials effectively mimic the surface tension controlling properties of currently marketed natural lung surfactants. Further, the polymer lung surfactants have strong protein resistance, which makes this class of materials promising also for potential use in Acute Respiratory Distress Syndrome (ARDS) treatment.

The application belongs to the technical field of antitumor drugs, and particularly relates to a nanofiber hydrogel for synergistic antitumor sonodynamic / chemodynamic therapy, a preparation method and application thereof, synthesis of Nap-Pept, preparation of Nap-Pept / Fe 3+ / Cur nanogel, under ultrasonic conditions, synergistic sonodynamic therapy of a traditional Chinese medicine sonosensitizer Cur and Fe 3+ mediated chemodynamic therapy, triggering immunogenic death of cancer cells by increasing ROS levels of B16F10 cells and then damaging mitochondrial function. Nap-Pept / Fe 3+ / Cur@US promotes expression of the immune-related factor calnexin, release of high mobility group protein 1 and maturation of dendritic cells to kill tumor cells, thereby exerting an antitumor effect. In vivo and in vitro studies show that the anticancer effect of the combined sonodynamic and chemodynamic therapy is superior to that of single therapy, achieving the purpose of precise and efficient anti-melanoma.

The invention discloses establishment and application of an epinephelus coioides frontal bone tissue osteoblast line, and relates to the technical field of biology. The cell line is an epinephelus coioides frontal bone tissue osteoblast line ECFBOB, the preservation number is CCTCC NO: C2025131, the cell line is derived from the frontal bone tissue of juvenile epinephelus coioides, and the cell line is established after tissue separation, primary culture by a dry pasting method, subculture and cryopreservation resuscitation. The obtained cell line can be used as an in-vitro research model, provides a key technical tool for saddletail grouper improved variety breeding and healthy culture in saddletail grouper skeletal development mechanism analysis, osteogenesis-related gene function verification, craniofacial bone malformation cause research and species traceability verification, fills the blank of saddletail grouper bone tissue cell lines, and has a wide application prospect. The method has important scientific research value and industrial application prospect.

The invention provides propionibacterium acnes and application thereof. Specifically, the invention provides the propionibacterium acnes B16, the propionibacterium acnes B16 is preserved in the China General Microbiological Culture Collection Center (CGMCC), and the preservation number is CGMCC No.31705. The propionibacterium acnes B16 has the advantages that the propionibacterium acnes B16 can be used for preparing the propionibacterium acnes B16; the strain B16 provided by the invention is separated and identified from pancreatic cancer excision tissues of clinical pancreatic cancer patients by adopting a bacterial separation and identification method. In-vitro research finds that propionibacterium acnes can significantly inhibit the growth of pancreatic cancer cells. Animal experiment results show that the caudal vein or intratumor injection can significantly inhibit the growth of pancreatic cancer. The experimental result shows that the B16 strain has the effect of inhibiting the growth of the pancreatic cancer, and prompts that the B16 has potential to become a microbial candidate drug for treating the pancreatic cancer.

The invention discloses a pentapeptide derived from cuttlefish ink and application thereof, and belongs to the field of biological medicine, the amino acid sequence of the pentapeptide is MINLP, and the potential function of the cuttlefish ink pentapeptide MINLP in preventing and treating metabolic syndrome is proved through in-vivo and in-vitro researches. And data support and theoretical support are provided for deeply researching the physiological regulation function of the marine medicinal biological resource cuttlefish ink polypeptide in the human body.

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 discloses a culture system and a culture method for intestinal organs of euryhaline fish, and belongs to the technical field of bioengineering. The culture system is composed of a basic culture medium and a complete culture medium, wherein the basic culture medium is Advanced DMEM / F12, and is supplemented with a GlutaMax, Hepes and mycillin double-antibody solution; and on the basis of the complete culture medium, N-2, B-27, N-acetylcysteine, EGF (Epidermal Growth Factor), Noggin, R-Spondin 1 and other key cell factor combinations are further added. By optimizing the tissue pretreatment and digestion process, the efficient fish intestinal tract organoid culture and passage method is established, the organoid model with the crypt-villus-like structure, the cell heterogeneity and the intestinal tract barrier function is successfully constructed, and the model provides a highly bionic in-vitro research tool for fish physiology and toxicology research.

The invention belongs to the field of biological medicines, and discloses a composition for treating immune drug-resistant colorectal cancer, the active components of the composition comprise a nano-drug CC (at) GE capable of inducing copper death and disulfide death and an immune checkpoint inhibitor, the nano-drug CC (at) GE comprises a nano-structure formed by coordination of copper ions and cystine, and the nano-structure is a nano-structure formed by coordination of copper ions and cystine. The nanostructure is coated with glucose oxidase and is loaded with tumor specific targeting peptide. In colorectal cancer treatment, cytotoxicity can be synergistically enhanced, tumor cell PD-L1 expression can be up-regulated, the anti-tumor immune effect of the PD-L1 inhibitor is enhanced, and therefore the treatment sensitivity of patients ineffective to the PD-L1 inhibitor is rebuilt. Experimental results prove that the composition has a remarkable anti-tumor curative effect in both in-vivo and in-vitro researches, and meanwhile, the nano-drug has high targeting to tumors, so that the nano-drug has low toxic and side effects in in-vivo treatment, has good safety and has a good development prospect in the field of biomedical application.

The invention discloses establishment and application of a humpback perch frontal bone tissue osteoblast line, and relates to the technical field of biology. The cell line is a humpback perch frontal bone tissue osteoblast line CAFBOB, the preservation number is CCTCC NO: C2025129, and the cell line is derived from a humpback perch juvenile fish frontal bone tissue and is established after tissue separation, primary culture by a dry pasting method, subculture and cryopreservation resuscitation. The biological characteristics are stable, 1-40 generations are mainly long fusiform, the chromosome karyotype is 2n = 48, strong alkaline phosphatase activity is achieved, the transfection efficiency is about 85%, and the activity is good after cryopreservation and resuscitation. The establishment method is high in repeatability, the obtained cell line can be used as an in-vitro research model and applied to morphological specialization mechanism analysis, osteogenesis-related gene function verification, classification dispute solution and seeding process research of the humpback perch, the blank of the bone tissue cell line of the humpback perch is filled, and the bone tissue cell line has important scientific research value and application prospect.

The invention discloses a method for detecting conformational change when protein recognizes a target site. The Cys at the 19 / 53 / 127 / 242 / 252 / 284 / 458 sites of the CasA protein in the Casade protein is mutated into Ser amino acid, and meanwhile, the 336 / 401 sites are mutated into the Cys, so that the obtained Cys-mini CasA keeps the function, and the mutant type of the CasA protein can be subjected to FRET fluorescence pair labeling, and therefore, the conformation state of the CasA can be truly reflected on the basis of single-molecule fluorescence resonance energy transfer smFRET, and the CasA can be used for detecting the CasA conformation. An effective means is provided for in-vitro research on the structure-function relationship of a system in CRISPR / Cascade, and the method has important significance for designing Cascade protein with optimized properties and enabling the Cascade protein to exert new functions.