141 results about "Disease modelling" patented technology

The invention discloses a Parkinson's disease animal model, a construction method and application, and relates to the field of animal genetic engineering and disease model construction. According to the model, seven mutation sites related to the human familial Parkinson's disease are introduced behind an initiation codon of a fourth exon of an endogenous SNCA gene, and the mutation sites are A18T, A29S, A30P, E46K, H50Q, G51D and A53T and have genotypes related to the human familial Parkinson's disease. The pig model shows nigra dopaminergic neuron reduction and cortical neuron reduction in the newborn period, and the core pathological process of the human Parkinson's disease can be systematically reproduced. The model can be widely applied to Parkinson's disease pathogenesis research, drug screening and cell therapy effect evaluation, and has important scientific research and preclinical application values.

The invention provides a human-derived recessive retinal degeneration disease organ-like model based on Prom1 gene knockout. Specifically, the invention provides a specific gRNA targeting a Prom1 gene exon 9, and the specific gRNA can be used for efficiently knocking out the Prom1 gene of the human embryonic stem cell. The invention also provides a Prom1 gene knockout stem cell line derived from the human embryo pluripotent stem cell H9 and a retinal organ disease model. The retina-like organ can provide a research model for clinical treatment drugs.

The application discloses a method for constructing a mouse model of atherosclerosis induced without high-fat diet, and relates to the technical field of disease model construction.The mouse model construction method provided by the application obtains a mouse model that spontaneously exhibits atherosclerosis symptoms without high-fat diet induction.The mouse model of atherosclerosis induced without high-fat diet can be used for researching the pathogenesis of atherosclerosis and screening and evaluating drugs for atherosclerosis.

The invention belongs to the technical field of animal model construction, and particularly relates to a non-alcoholic fatty liver disease mouse model based on humanized PNPLA3 I148M transgene as well as a construction method and application thereof, the construction method comprises the following steps: step i: designing CDS of a human gene and a key 3 'UTR element for patent medicine of a small nucleic acid drug in gene plasmid DNA of the constructed model; step ii, using Luc2 or iRFP to report gene expression in the gene plasmid DNA for constructing the model; and (iii) a disease model of the fatty liver disease induced by combining plasmid DNA transient transfer with high-fat diet. The mouse model constructed by the invention has the advantages that a specific exogenous gene expression signal of the liver is detected in real time through small animal living imaging, a human PNPLA3 protein is detected and overexpressed through serum ELISA, PNPLA3 I148M mRNA is detected and overexpressed through liver RT-qPCR, the phenotype of the non-alcoholic fatty liver disease of the liver is determined through 15-week-old pathology, and the whole process of modeling for 8 weeks does not need to be operated in a biosafety secondary laboratory.

The invention discloses a reverse transcriptase, wherein a coding sequence of the reverse transcriptase is derived from a Rattus norvegicus genome. The protein sequence of the protein comprises a protein sequence of a wild type Rattus norveicus, or a protein sequence which is subjected to engineering modification on the basis of the wild type Rattus norveicus, or a protein sequence which is subjected to engineering modification on the basis of the wild type Rattus norveicus. Furthermore, the protein sequence of the gene is as shown in any one of SEQ No.1-55. In a rice genome site and a human HEK 293T cell genome site, the system shows efficient editing efficiency of a guide editor. The invention provides a series of efficient reverse transcriptase components which can be carried on a guide editing system, important bottom-layer technical support is provided for research of point mutation in a genome, creation of a special site disease model and correction of genetic mutation sites, and the reverse transcriptase component has a wide application prospect in the field of precision medicine and precision agriculture breeding.

The invention belongs to the field of biosensing and fluorescence imaging, and particularly relates to a fluorescent probe compound and application thereof. The probe compound is a compound as shown in a formula I or a compound as shown in a formula II, or a pharmaceutically acceptable salt thereof; the structures of the compound in the formula I and the compound in the formula II are shown in the specification, and the definition of each group is shown in any embodiment of the invention. The fluorescent probe compound shows rapid and high reactivity to oxidizing free radicals, the generated fluorescent compound is high in fluorescence intensity and good in stability, and early diagnosis can be carried out on ferroptosis-related disease models such as oxygen-glucose deprivation / reperfusion and drug liver injury on living cell or living body levels and the like. I II

The present invention relates to a biomarker composition for diagnosing stroke or post-stroke neurological damage, for diagnosing the severity of stroke or post-stroke neurological damage, and for predicting stroke prognosis, and the like. Using an ischemic stroke mouse disease model, a tissue damage analysis and a behavioral analysis were performed, and the mRNA level expression was confirmed, and as a result, Prox1 or Dcx could be selected as a biomarker. In addition, it was confirmed that the expression of Prox1 or Dcx increased as the severity of stroke or post-stroke neurological damage was higher, and thus the present invention is expected to be useful for diagnosing stroke or post-stroke neurological damage, for diagnosing the severity of stroke or post-stroke neurological damage, and for predicting stroke prognosis.

This invention relates to a method for constructing a mouse orthotopic model of highly aggregated glioblastoma, belonging to the fields of biomedicine and experimental animal model technology. Glioblastoma cells are resuspended in a composite carrier of a specific ratio of matrix gel and serum-free culture medium, and a low-temperature injection technique is used. Utilizing the thermosensitivity of matrix gel in the mouse intracranial environment (i.e., its phase transition from liquid to gel at a certain temperature), precise colonization and physical locking of tumor cells at target coordinate points are achieved. This method improves upon existing glioblastoma mouse orthotopic model construction techniques that suffer from tumor cell diffusion and loss, low tumor formation rate, irregular tumor morphology, large intragroup variability, and needle reflux due to low viscosity of the inoculation carrier. It provides a good disease model for subsequent screening or evaluation of anti-glioblastoma drugs.

The invention belongs to the technical field of disease model construction, and particularly relates to a construction method of a chronic myelogenous leukemia mouse model with conditional human EZH2 overexpression and RUNX1 knockout. According to the invention, a chronic myelogenous leukemia mouse transgenic mouse model with conditional human EZH2 overexpression and RUNX1 knockout is successfully constructed, the model is induced to be converted from a chronic stage to a sudden change stage, and particularly, the model is a transgenic mouse model which is positive in Lyz2-CreERT2 / EZH2 / RUNX1 and carries BCR-ABL and SCL-tTA. It is proved that a human EZH2 conditional overexpression and RUNX1 knockout chronic myelogenous leukemia mouse transgenic mouse model has feasibility and importance for research on conversion from CML CP to BC samples, and a molecular mechanism for conversion from chronic myelogenous leukemia to a sudden change stage is revealed for research. And a new animal model and a new research idea are provided for understanding of disease progression and development of a new treatment strategy.

A method for the selection of processing parameters for nanomaterial compositions is performed by preparing a baseline model wherein a nanomaterial composition interacts with a disease model. It is then acquired a baseline interaction between the nanomaterial composition and the disease model. A perturbation is applied iteratively by changing at each iteration at least one parameter of the perturbation and acquiring at each iteration a perturbation parameter describing a protein corona formation on said first nanomaterial composition under the effect of the perturbation. Finally, it is identified and selected among the perturbation parameters at least one processing parameter minimizing the formation of protein corona.

The invention discloses a method for constructing astrocytes serving as a smoke disease model, and belongs to the technical field of crossing of stem cells and neuroscience. The method comprises the following steps: S1) reprogramming CD34 + cells in in-vitro PBMCs (peripheral blood mononuclear cells) of smoke disease patients carrying and not carrying RNF213p.R4810K mutation to obtain induced pluripotent stem cells; and S2) directionally inducing and differentiating the induced pluripotent stem cells into astrocytes through a neural progenitor cell way, wherein the obtained astrocytes are the astrocytes capable of being used as the smoke disease model. The astrocyte model prepared by the invention can be used for researching pathogenesis, nerve-blood vessel interaction process and blood-brain barrier (BBB) related functions of smoke diseases, and can be further applied to molecular typing of diseases and in-vitro function evaluation of candidate drugs.

The invention belongs to the technical field of gene editing and disease model construction, and particularly relates to a construction method and application of a Wilson disease animal model. Aiming at a large fragment deletion mutation type which exists clinically in Wilson disease but lacks a corresponding animal model, a CRISPR / Cas9 gene editing technology is utilized, a pair of sgRNAs is specifically designed, an eighth exon region of a mouse Atp7b gene is precisely cut and deleted, and a WD mouse model with deletion mutation (c.2333340delGACGGTGG) of eight basic groups of the eighth exon of the Atp7b gene is successfully constructed. The method can be used for research on pathogenesis of WD, screening of novel diagnostic markers and evaluation of curative effect of therapeutic drugs, is particularly suitable for preclinical evaluation of gene therapy strategies and development of adjuvant therapeutic drugs of targeted NLRP3 inflammasomes, and has great scientific research value and clinical transformation prospect.

The application provides a reprogramming vector for blood cells and application thereof. Through the technical scheme in the disclosure, a low-cost and high-benefit solution can be provided for efficient reprogramming and application of iPSCs, and the wide application of iPSCs in clinical and scientific research fields is significantly promoted, and the iPSCs have important application value and development potential in research and practice in aspects such as disease models, drug screening, cell therapy and the like.

The invention relates to the technical field of organoids, and particularly discloses a chick intestinal tract crypt stem cell separation medium and a separation method and application thereof, and the technical key points are as follows: the separation medium provided by the scheme of the invention can efficiently separate chick intestinal tract crypts with complete structure, good activity and high purity under a low-temperature condition by optimizing component combination; according to the separation method provided by the scheme of the invention, the separation efficiency and the culture quality of the intestinal stem cells and the intestinal organs are remarkably improved through the matching of the separation liquid and the washing liquid. By utilizing the method disclosed by the invention, a high-quality stem cell source can be provided for culture of intestinal organs, drug screening and disease model construction, and the method has important scientific research and application values.

The present invention provides methods of differentiating pericytes from pluripotent stem cells comprising culturing steps in chemically defined culture medium. A population of exogenously derived pericytes from PSCs are also contemplated. Further uses of the exogenously cultured pericytes for an in vitro disease model or in vitro angiogenesis assay are contemplated, including an in vitro 3D model of vasculature.

The invention discloses an intelligent cerebral apoplexy cell state recognition method based on Cellpose and fluorescence image analysis and application of the intelligent cerebral apoplexy cell state recognition method in drug effect screening. The intelligent cerebral apoplexy cell state recognition method comprises the following steps: S1, establishing a cerebral apoplexy ischemia reperfusion cell model; s2, adding a nitrite fluorescent probe into the cells subjected to ischemia reperfusion treatment to obtain a fluorescent microscopic image of the cells; s3, importing the fluorescence microscopic image into a Cellpose model for cell segmentation, and obtaining an ROI region of each cell; s4, extracting fluorescence intensity characteristics and cell morphological characteristics in the ROI region of each cell; and S5, inputting the extracted multi-dimensional features into a classification model for cell state recognition, so as to distinguish a single cell into three states of health, mild injury or serious injury. According to the method, a complete link of image acquisition-Cellpose analysis-feature extraction-state discrimination-drug effect evaluation is constructed, and the practical application value of a deep learning image algorithm in disease model research and drug screening scenes is expanded.

The invention discloses a novel ALS disease model and application thereof in screening ALS treatment drugs, and belongs to the technical field of disease models and drug screening. According to the method, induced pluripotent stem cells derived from a sporadic ALS patient are directionally induced into spinal cord organs, the spinal cord organs and immune cells derived from the same donor form a co-culture system, the co-culture system is infected with viruses, and the ALS disease model is successfully constructed. According to the method, ALS diseases are simulated on the three-dimensional level, ALS disease models of all stages (including the initial stage of the diseases) are obtained, and the pathological features of all the disease stages of ALS (especially 90% or above of sporadic ALS) can be accurately reflected; the problem that key disease-promoting factors in the early stage of the disease cannot be obtained due to complete disorder of downstream signal channels in the end stage of the disease after definite diagnosis caused by slow definite diagnosis of hidden disease onset of the ALS is solved.

The present disclosure relates to a dwarfism animal model carrying an IGF-1 gene mutation and a method for generating the same. According to the present disclosure, the problem that an animal dies immediately after birth is overcome, the majority of phenotypes seen in Laron syndrome patients may be observed in the dwarfism animal model, and the dwarfism animal model has decreased expression of personality genes. Thus, the dwarfism animal model may be effectively used as a dwarfism-related disease model.

Provided is a structure composed of a cell population comprising endothelial cells, astrocytes and pericytes, and a 3D (three dimensional) cell growth material within which the cell population is located. The structure has a TEER value of at least 450 Ω / cm2. The cells of the structure may be derived from the brain. The cells may be human cells, and in particular may be primary derived non-immortalised cells. The structure is particularly suited for use in a model of the blood brain barrier, and the invention also provides such a model. The structure is located in a container, in which it separates a first chamber located on a first side of the structure and a second chamber located on a second side of the structure. The first and second chambers respectively contain first and second liquids in contact with first and second sides of the structure. The liquids mimic the brain extracellular fluid and the blood. The blood brain barrier model provided may be used in models of brain disease, and to investigate uptake of agents into the brain or diseased brain.

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 application belongs to the field of bioengineering and gene editing technology, and particularly relates to a TIGR-Tas cytosine base editing system and application thereof. The application provides a PAM-independent, efficient controllable and low byproduct TIGR-Tas cytosine base editor and an optimized variant thereof by fusing cytosine deaminase with a TIGR-Tas system effector protein parTasR and combining mismatch programming of tigRNA to induce parTasR to be in a nickase-like catalytic state, so as to realize windowed C→T base conversion in mammalian cells and provide better technical support for disease model construction and gene therapy and the like.

The present disclosure relates generally to providing a cellular time-series imaging, modeling, and analysis platform, and more specifically to acquiring time-series image data and using various machine learning models to model and analyze subcellular particle movements and changes in cellular positional and morphological characteristics using unsupervised embedding generation. The platform can be applied to evaluate various cellular and subcellular processes by generating summary embeddings of time-series image data that enable analysis of dynamic cellular and subcellular processes over time (e.g., the movement of particles within a cell, neurites on developing neurons, etc.) for enhanced identification of differences between cell states (e.g., between sick and healthy cells) and generation of disease models which can be used to analyze the impact of various therapeutic interventions, among other improvements described throughout.

The invention relates to the field of gene vector construction, in particular to construction and application of a gene vector integrating fluorescent screening and self-deletion functions, the gene vector comprises a conditional knock-out (cKO) vector and a gene knock-in (KI) vector which are both integrated with a Dre-Rox mediated self-deletion module and a fluorescent screening module, the construction method comprises the following steps: carrying out HindIII / EcoRI double enzyme digestion on a pUC19 vector, carrying out homologous recombination amplification on a target fragment, carrying out connection transformation and screening verification to obtain a qualified vector; the vector is applied to preparation of cKO / KI gene modified mice, and efficient screening of positive individuals is realized through fluorescence preliminary screening, genotype identification and fluorescence quenching detection. The positive screening workload and cost can be reduced by 70% or above, non-target elements are accurately cut off, interference is avoided, time-space accurate regulation and control are achieved, carrier construction is easy and convenient, repeatability is high, and the method is suitable for gene function research, disease model construction and drug target verification.

The invention provides a probiotic strain formula scheme screening method. The method comprises the following steps: establishing a disease model database according to a clinical examination result, performing standardization processing and data layering on the disease model database to obtain a layered database, scoring and grading diseases in the layered database, and establishing a disease model grading data matrix of patients; establishing a strain curative effect matrix according to the strain database and published clinical research data; and associating the disease model grading data matrix with the strain curative effect matrix, screening a relationship with high association strength, constructing a disease-microbial treatment response model, and obtaining different types and different levels of diseases and corresponding recommended bacterial systems through the disease-microbial treatment response model. And grading the synergistic effect of the disease and the recommended strain, filtering the score through a threshold value to screen out a scheme of a strain formula, and outputting the scheme of the strain formula through a user interface.

The present application relates to the technical field of induced pluripotent stem cell, and particularly relates to mesoderm differentiation specific human induced pluripotent stem cell and application thereof. The present application provides human induced pluripotent stem cell BC-hiPSC-ME-20, which is preserved in China General Microbiological Culture Collection Center, and the preservation number is CGMCC No.46536. The cell strain has multi-lineage differentiation potential and high-efficiency mesoderm directional differentiation capacity, has stable pluripotency and excellent passage stability, and has higher safety, and has a good application prospect in disease model construction, disease occurrence and development mechanism research, drug screening and cell therapy.

The invention discloses a 15-antibody combination for accurately detecting a mouse tissue immune cell subset and application of the 15-antibody combination. Aiming at the problems of large fluctuation and instability of detection results caused by adjustment of voltage and fluorescence compensation during detection by a flow cytometer and different processing modes during result analysis, the antibody combination disclosed by the invention takes monoclonal antibodies CD45, CD3, CD4, CD8, CD19, B220, Ly-6C, Ly-6G, MHC-II, CD11c, CD49b, CD62L, CD44 +, PD-1 and PD-L1 as effective components, and a stable and standard detection method for mouse tissue immune cell subsets is established. The method can analyze the proportion, number and distribution of various immune cells (T cells, B cells, NK cells, dendritic cells, mononuclear cells and granulocytes) in spleen, lung, liver, brain or kidney tissues of mice, can reflect the immune state of the mice and evaluate the immunosuppression state, and can be applied to the fields of disease models, drug evaluation, vaccine development and the like.

The invention discloses an NDUFS2 gene heterozygous editing pig as well as a construction method and application thereof, and relates to the field of animal gene engineering and disease model construction. The eighth exon of the NDUFS2 gene of the pig is deleted or mutated, and presents an NDUFS2 heterozygous genotype; in the newborn period, the number of neurons of the middle cerebral cortical layer and the number of dopaminergic neurons are obviously reduced, the number of neurons of the adult cortical layer is continuously reduced along with mitochondrial swelling and crest fracture, and after MPTP treatment, the number of neurons of the adult cortical layer is disordered in motion trail, and the body tremor index is increased. The pig model not only can be used for revealing the action mechanism of the NDUFS2 defect in neurodegenerative diseases (such as Parkinson's disease), but also can be used as an important tool for screening new drugs, evaluating treatment means and researching mitochondrial diseases.

The invention discloses a state recognition method of an adaptive gene editing laboratory mouse, and aims to solve the problem of insufficient state recognition specificity of the gene editing laboratory mouse in the prior art. The method comprises the following steps: acquiring genotypes, disease model types and experimental environment parameters of laboratory mice; optimizing detection and optical flow analysis parameters, generating a time sequence behavior track map and segmenting a key area; extracting general and pathological specific composite features and standardizing the general and pathological specific composite features; calling a genotype pathological feature association database to calibrate a weight and a threshold value, and inputting a random forest classifier recognition state of transfer learning optimization; scores are calculated through a weighted scoring model, the state is judged, and credibility is calculated through a multi-dimensional confidence evaluation model. The method is suitable for various gene editing laboratory mice and experimental environments, the accuracy and suitability of state recognition are improved, and powerful support is provided for gene therapy research and development and disease mechanism research.

This invention discloses a skin organ-on-a-chip, relating to the fields of biomedical engineering and microfluidics. The chip includes a drive module integrating a reservoir with a fixed height difference, a culture layer with arrayed independent skin culture units, and a fluid layer separated by a porous membrane. The drive module utilizes the hydrostatic pressure generated by the liquid level difference to drive the culture medium to form a stable laminar flow in an optimized fluid channel network, achieving pump-free perfusion. This invention achieves stable pump-free drive through a reservoir with a fixed height difference; the integrated sealed structure eliminates the risk of contamination and ensures complete isolation between units; the simulation-optimized uniform flow channel network, in synergy with the drive, ensures high consistency in high-throughput parallel culture, enabling the chip to provide a long-term stable biomimetic perfusion microenvironment. A single experiment can efficiently complete multiple replicates and multi-condition transdermal drug screening, toxicity testing, and disease model construction, improving experimental throughput, data reliability, and ease of operation.

A method is provided for predicting a disease outcome using a machine learning model that generates training data for training the machine learning model useful for implementing a cellular disease model.SOLUTION: A method for predicting a disease outcome using a machine learning model includes implementing an ML-enabled cellular disease model to validate an intervention, identifying a patient population likely to be a responder to the intervention, and developing a therapeutic structure activity relationship screen. To generate a cellular disease model, data from human genetic cohorts, the literature, and generic cellular or tissue-level genomic data are combined to elucidate a set of factors (e.g., genetic, environmental, cellular factors) that cause a particular disease. A series of factors are used to manipulate invitro cells to generate training data for training a machine learning model useful for implementing a cellular disease model.SELECTED DRAWING: FIG. 1B