4 results about "Human myocardium" patented technology

The application discloses an antibody for detecting cardiovascular diseases and application thereof, and belongs to the technical field of biological medicine. The monoclonal antibody or antigen binding fragment thereof provided by the application can specifically bind to a specific antigen epitope (SEQ ID NO:1) of human cardiac troponin I (cTnI) and has no cross reaction with skeletal muscle troponin I. The application further discloses the CDR sequences of the heavy chain and light chain variable regions of the antibody, a coding gene, a preparation method and application thereof. An ELISA detection kit containing the antibody can specifically recognize cTnI in a blood sample, has good linearity in the range of 0.01-50 ng / mL, has a false negative rate of 0 for serum detection of patients with acute myocardial infarction, and has a low false positive rate for serum detection of healthy people. The antibody and the kit have important application value in rapid and accurate diagnosis of cardiovascular diseases such as acute myocardial infarction.

The invention relates to the field of cell engineering, and particularly discloses a preparation method and application of a conditionally immortalized human myocardial fibroblast model. According to the method, a Tet-on system driven by a cardiac fibroblast specific promoter is constructed to regulate and control a lentiviral vector of an SV40LT-tsA58 temperature-sensitive mutant gene, and after primary human cardiac fibroblasts are transduced, conditional immortalization of the cells is realized in the presence of an inducer doxycycline. The model can be stably amplified under an induction condition, the immortalized gene expression is closed after an inducer is removed, the primary physiological state of cells is recovered, and the high-fidelity response capability to fibrosis promoting and anti-fibrosis drugs is reserved. According to the method, the in-vitro proliferation limit of primary cells is broken through, the problems of species difference and ethics are avoided, and a humanized, high-fidelity and high-controllability standardized tool is provided for myocardial fibrosis pathological mechanism research and drug screening.

The invention provides a construction method of a ZBED6 gene-deleted human myocardial cell line, which comprises the following steps: designing and synthesizing two sgRNAs specifically targeting a human ZBED6 gene coding region, respectively cloning the sgRNAs into a pSpCas9 (BB)-2A-Puro vector, constructing a recombinant expression vector, then transfecting the recombinant expression vector into a human AC16 myocardial cell, and carrying out screening, monoclonal separation and culture to obtain the ZBED6 gene-deleted human myocardial cell line. The human myocardial cell line with the ZBED6 gene deletion is obtained. The ZBED6 gene deleted human myocardial cell line constructed by the invention has a stable cell hypertrophy phenotype, and can be used as a myocardial hypertrophy in-vitro model for screening and preparing myocardial hypertrophy related drugs. The ZBED6 gene deleted human myocardial cell line constructed by the invention can realize continuous passage and maintain a stable ZBED6 deletion state, and by establishing a puromycin gradient screening strategy and optimizing a monoclonal separation technology, the positive cloning rate is effectively improved, cross contamination and cell damage are avoided, the uniformity and stability of a cloning genetic background are ensured, and the application prospect is broad. Wide application prospects are realized.

The invention discloses a high-throughput screening method for anti-myocardial injury activity of catechin compounds and application of the high-throughput screening method. The method is based on an AC16 human myocardial cell strain, and isoprenaline is adopted for inducing and establishing a standardized myocardial injury model; by integrating cell activity detection, apoptosis analysis, PI3K / AKT signal channel activity monitoring, myocardial injury marker quantification and cell morphology intelligent image analysis, a multi-parameter comprehensive evaluation system is established, an image analysis algorithm and a comprehensive activity scoring model based on deep learning are developed, a catechin compound structure-activity relationship database is constructed, and a catechin compound structure-activity evaluation model is established. The method has the advantages that activity prediction is realized, a liquid treatment workstation, a high-content imaging system and other automatic equipment are integrated, 480-1920 compound samples can be evaluated in a single experiment, the screening period is shortened to 72-96 hours, the screening efficiency is improved by 10-20 times, and the method is suitable for rapid screening and action mechanism research of myocardial protection drugs.