110 results about "In vitro study" patented technology

The invention relates to the field of biology, in particular to a biological material for overexpressing a chicken lnc-CA13 gene and application of the biological material. The chicken lnc-CA13 gene provided by the invention is inserted into a lentiviral vector, a biological material (recombinant vectors (lentiviral recombinant vectors, lnc-CA13-4) and recombinant bacteria) capable of improving the expression of the chicken lnc-CA13 gene can be obtained, the biological material can effectively improve the expression quantity of the chicken lnc-CA13 gene, and the overexpression efficiency at the gene level is 95.85%. The chicken lnc-CA13 gene is constructed on a lentiviral vector and is suitable for in-vivo and in-vitro research; meanwhile, the invention has important scientific significance and application prospects in the aspects of deeply researching the functions of the chicken lnc-CA13 gene and the like.

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 belongs to the technical field of immunology, and particularly discloses an in-vitro preparation method and application of an induced follicular regulatory T cell iTfr.The in-vitro preparation method comprises the steps that mouse spleen single-cell suspension or healthy human peripheral blood is obtained, peripheral blood mononuclear cells are obtained, then initial CD4 + T cells are obtained through immunomagnetic bead sorting, and iTfr cells are obtained; then carrying out induced differentiation by using an antibody and a specific cytokine, and carrying out continuous stimulation and culture by adopting two different cytokine polarization conditions, so as to obtain the iTfr cell. The invention provides a method for preparing the iTfr cells in vitro, which is expected to obtain an enough number of induced follicular regulatory T cells, and provides an important tool and a treatment means for in vitro research of immune tolerance related diseases such as autoimmune diseases and organ transplantation.

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.

The invention provides a mutant spinal glioma cell line. The H3K27M mutant type glioma cell line is successfully constructed, the constructed cell line has more excellent multiplication capacity, migration capacity, tumor formation capacity and drug resistance, the curative effect and potential side effects of drugs can be more comprehensively evaluated, the cell line can be used for in-vitro research on the biological function of H3K27M glioma and response to drug treatment, and the cell line can be applied to in-vitro research of the H3K27M mutant type glioma. Wide application prospects are realized.

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.

The invention discloses application of carboxamidotriazole combined chemotherapeutic drugs in preparation of drugs for treating solid tumors, which mainly aims at the chemotherapeutic drugs such as gemcitabine, docetaxel or albumin paclitaxel to achieve the technical effects of effect enhancement and toxicity reduction, and the solid tumors are lung cancer, breast cancer, liver cancer and kidney cancer. In-vivo and in-vitro researches prove that the combination of carboxamidotriazole and gemcitabine, docetaxel or albumin paclitaxel can achieve a remarkable combined synergistic effect, and does not increase toxic and side effects, so that the carboxamidotriazole can benefit more patients through clinical combined medication.

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 technical problem to be solved by the invention is to provide the ovarian cancer tumor immune microenvironment in-vitro model based on the micro-fluidic chip and the use method. The in-vitro model is beneficial to systematic understanding of individual effects and synergistic effects of all components in tumor progression, and can also guide use of related drugs and predict drug use effects. The in-vitro model comprises a chip body, at least one culture medium flow channel is formed in the left end face of the chip body, at least one group of organ-like holes are formed in the upper surface of the chip body in the left-right direction of the chip body and communicated with the culture medium flow channel under the organ-like holes, a Transwell chamber is embedded in each organ-like hole, and the in-vitro model further comprises a culture medium injection device; compared with a traditional co-culture model, the chip model is closer to an in-vivo real condition and has time priority in the aspect of immune response change; in-vitro study on the unique tumor immune microenvironment of the ovarian cancer is facilitated, and meanwhile, the change condition of the tumor immune microenvironment after in-vivo drugs are used is also facilitated.

The invention discloses an acute myelogenous leukemia cell strain with multiple adverse prognostic gene mutations in human and application thereof, the cell strain is named as human acute myelogenous leukemia cell strain YYXY-M14 and preserved in China Center for Type Culture Collection (Wuhan University, Wuhan, China) on October 12, 2024, and the preservation number is CCTCC NO: C2024350. The acute myelogenous leukemia cell strain simultaneously has three acute myelogenous leukemia poor prognostic gene mutations of TP53, RUNX1 and ASXL1, has the in-vitro natural proliferation capacity of acute myelogenous leukemia cells, and can be used as a cell material for researching the pathogenic mechanism of human acute myelogenous leukemia with positive mutation of human TP53, RUNX1 and ASXL1 in vitro. Meanwhile, the method can be used for in-vivo animal model construction and acute myelogenous leukemia drug screening and evaluation research.

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 the field of biology, in particular to shRNA for inhibiting chicken lnc-CA13 gene expression, a recombinant vector, recombinant bacteria and application of the shRNA. The invention provides shRNA (short hairpin Ribonucleic Acid) for inhibiting expression of the chicken lnc-CA13 gene, and constructs a corresponding shRNA lentiviral expression vector and recombinant bacteria. The nucleotide sequences of the three shRNAs provided by the invention are shown as SEQ ID NO.5-10, the three shRNAs can effectively reduce the expression quantity of the chicken lnc-CA13 gene, and the inhibition efficiencies at the gene level are respectively 27.88%, 54.58% and 39.42%. ShRNA for inhibiting chicken lnc-CA13 gene expression is constructed on a lentiviral vector, and the lentiviral vector is suitable for in-vivo and in-vitro research; meanwhile, the invention has important scientific significance and application prospects for deeply researching the functions of the chicken lnc-CA13 gene and the like.

The invention discloses human laryngeal squamous cell carcinoma related fibroblasts, which are classified and named as human laryngeal squamous cell carcinoma related fibroblasts GY-HNSCC-CAF Homo sapiens, the preservation unit is China Center for Type Culture Collection, the preservation number is CCTCC NO: C2024179, and the preservation date is July 7, 2024. The invention enriches the resource library of tumor-related fibroblasts, especially fibroblasts from head and neck squamous cell carcinoma, can be used as an effective in-vitro study cell model, and provides a new experimental model for the purposes of etiology, metastasis mechanism, drug resistance mechanism, drug screening and the like of head and neck squamous cell carcinoma.

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 belongs to the field of medicine product manufacturing, relates to manufacturing and quality control of inhalation preparation medicine products, and provides a drug delivery particle collecting device used in inhalation preparation in-vitro research and evaluation. By optimizing the structure of the device, the provided drug delivery particle collecting device is not only suitable for collecting and measuring the traditional delivery dosage, but also especially suitable for applying the delivered drug particles to in-vitro dissolution evaluation; the medicine particles collected by the device provided by the invention can still keep extremely high dissolution curve similarity under different medicine deposition amounts (the similarity factor f2 of the optimal device can reach 90.6). On the basis, the influence of the medicine deposition amount on the dissolution rate can be avoided, and the product quality of the inhaled preparation medicine can be evaluated more objectively, so that the product research and development and quality control of the inhaled preparation new medicine or the imitated medicine can be smoothly promoted.

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.