10 results about "Human myocardium" patented technology

The invention discloses a nano antibody targeting human cardiac troponin I and application of the nano antibody. According to the invention, through construction of a nano-antibody phage library and a phage screening technology, four nano-antibodies targeting human cardiac troponin I and having a good pairing effect are obtained. Compared with the traditional monoclonal antibody, the bacteriophage display nano antibody has the advantages of small size, high stability, simplicity and convenience in preparation, low cost, large-scale production and the like; the probe can be used as a core detection element and is applied to the field of diagnosis of clinical rapid detection of human cardiac troponin I.

The invention discloses a phosphorylation-inhibited AKT1 gene mutant type human myocardial microvascular endothelial cell line as well as a construction method and application thereof. The 473th amino acid of an AKT1 gene in the cell line is mutated from serine to alanine (S473A). The phosphorylation-inhibited AKT1 gene mutant type human myocardial microvascular endothelial cell line is successfully constructed, the phosphorylation-inhibited AKT1 gene mutant type human myocardial microvascular endothelial cell line has stable genetic characteristics, progeny cells can be continuously used for research, repeated cell culture and gene editing operation is avoided, and time and experiment cost are remarkably saved. The invention provides a stable cell model, effectively reduces the batch-to-batch difference caused by the traditional instantaneous editing technology, provides a reliable and repeatable experimental tool for revealing the specific mechanism of diabetic myocardial microvascular injury, improves the research accuracy, and has a wide application prospect. And a solid theoretical basis and an experimental basis are laid for developing a treatment strategy aiming at the AKT1 target spot.

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 application provides application of a calcium ion indicator GCaMP in detection of a calcium signal of a human myocardial cell line, successfully applies the calcium ion indicator GCaMP to detection of a calcium signal of a myocardial cell, and increases the use of the calcium ion indicator GCaMP in detection of a calcium signal of a myocardial cell. Secondly, the application provides a method for detecting a calcium signal of a myocardial cell based on the calcium ion indicator GCaMP, and detecting agonism and / or inhibition of a myocardial cell PIEZO1 channel by a small molecule compound, and lays a foundation for research on a myocardial cell related calcium channel.

The invention belongs to the technical field of bioelectrochemical detection, and provides a surface modified electrode and a preparation method and application thereof, and the preparation method specifically comprises the following steps: (1) carrying out surface assembly on a gold electrode by using a thiohydracrylic acid solution to obtain a monomolecular layer electrode; (2) mixing the activation solution and a monomolecular layer electrode for activation to obtain an activated electrode; (3) culturing the monoclonal antibody solution and the activated electrode to obtain a stable electrode; and (4) mixing a phosphate solution and the stable electrode, and sealing to obtain the surface modified electrode. A step-by-step modification system of self-assembly of a monomolecular layer, activity activation, antibody immobilization and closing is constructed, a gold electrode is used as a substrate, and the surface of the gold electrode is sequentially modified with a mercaptopropionic acid self-assembly monomolecular layer and a mouse anti-human cardiac troponin I (cTnI) monoclonal antibody immobilized through a covalent bond; and the bovine serum albumin layer is used for sealing non-specific sites, so that a stable biological recognition interface is formed.

The invention is applicable to the technical field of biological medicines, and provides application of an HIF-2alpha specific inhibitor in preparation of a medicine for treating diabetic cardiomyopathy. The invention clarifies the copper death of human myocardial cells induced by combination of high glucose and high fat and copper and related molecular mechanisms, namely when the extracellular copper concentration is increased, copper transporter CTR1 is reduced, DMT1 compensatory is increased, copper is transported into AC16, HF + HG further promotes DMT1 expression by enhancing m6A methyltransferase WTAP expression to mediate HIF-2alpha m6A modification, and finally the copper ion concentration in AC16 is increased, so that the copper death of the human myocardial cells is induced. In order to solve the problem that the copper death is caused due to the fact that the HIF-2 alpha specific inhibitor PT2385 is used for inhibiting the HIF-2 alpha, the expression of DMT1 in AC16 cells is reduced, the copper death index is recovered to be normal, and then a new target is provided for treating DCM.

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.

The invention discloses application of circular RNA circRftn1 in diagnosis of cardiac hypertrophy. According to the application, research finds that the circular RNA circRftn1 is highly expressed in cardiac muscle tissues of a cardiac hypertrophy model mouse and is highly expressed in peripheral blood and cardiac muscle tissues of a cardiac hypertrophy patient; circRftn1 is a circular RNA (Ribonucleic Acid) which is found for the first time; the circular RNAcircRftn1 in the tail venous blood of the mouse can be used as a diagnostic marker of the cardiac hypertrophy of the mouse; the circular RNA circRftn1 in human peripheral blood can be used as a diagnostic marker of the human cardiac hypertrophy, and the circular RNA circRftn1 can be used for early large-scale screening of the human cardiac hypertrophy.