71 results about "Bronchial Epithelial Cell" patented technology

The invention discloses a method for rapidly separating and culturing human bronchial epithelial cells, and an optimized medium. The optimized medium is prepared from a DMEM culture medium, an F-12 Nutrient mix culture medium, 0.5 mg/ml hydrocortisone, fetal bovine serum, 25mu g/ml epidermal growth factor, 5 mg/ml insulin, 11.7mu M cholera toxin, 10 mM ROCK1 inhibitor, and 10 mM BMP4 antagonist. The method for rapidly culturing the human bronchial epithelial cells by using the optimized medium not only greatly simplifies the steps of culturing epithelial cells which require 3T3 J2 cells as a feeder layer, and only needs to culture the primary human bronchial epithelial cells digested by pronase in the optimized medium, but also can be used for obtaining humane bronchial epithelial cells which are rapidly separated and can be indefinitely subcultured; the subcultured human bronchial epithelial cells are subjected to in vitro expansion and culture; when the human bronchial epithelial cells are subcultured for 20 generations or more, a sufficient amount of seed cells can be obtained so as to be used in a variety of follow-up in vitro and in vivo studies such as in vitro cell interaction model studies.

The invention discloses a human normal bronchial epithelial cell and preliminary isolated culture and subculture methods and purposes of the human normal bronchial epithelial cell. The cell is named as the human normal bronchial epithelial cell HNBEC/HL-001, the preservation number of the cell is CCTCCNO: C201311. The preliminary isolated culture method includes the steps that fat in a para-carcinoma tissue sample excised from a patient with lung cancer through an operation is removed, dispase and DNasel are added to the fat for action after the fat is digested, the cell is collected through filtering and centrifugal operations, and the cell is suspended again with an HL culture medium for inoculated culture. The subculture method includes the steps that when the cell proliferates to 70-90% abundance, pancreatin-EDTA is used for digestion, and then DMEM is used for neutralization; the cell is collected through a centrifugal operation, and the cell is suspended again with an HL culture medium for inoculated culture. The human normal bronchial epithelial cell can be used for physiological research of human normal cells, drug toxicity research and detection of external normal cells, and research on pathogenesis of bronchia and lung diseases including bronchogenic carcinoma.

The invention relates to a novel application of formononetin in the medical science, in particular to the application of formononetin in preparing medicine for preventing and treating airway inflammation and asthma. Pharmacological test result shows that the formononetin participates in reducing the allergic airway inflammation of model animals/occurring and developing of asthma lesion; reducing the of damage inflammatory factor to the human bronchial epithelial cell in vitro and the formononetin can be used for preparing the medicine for treating airway inflammation diseases, such as asthma, bronchitis, capillary bronchitis and air or non-air idiotoxin or inflammatory or non-inflammatory of irritant; and specifically, the formononetin has the application in preparing medicine for treating allergic airway inflammation and asthma disease.

The present invention is for an isolated peptide consisting of the amino acid sequence of SEQ ID NO: 9 and using the peptide for treating asthma by reducing cytotoxicity of eosinophil derived toxins in bronchial epithelial cells of the subject suffers from asthma. The method comprises preparing a pharmaceutical composition having the peptide of SEQ ID NO: 9 and administering an effective amount of the composition to the subject.

The invention discloses an extraction method of glycyrrhiza polysaccharide and application. The glycyrrhiza polysaccharide is prepared by combining a high temperature organic acid method and ultrafiltration membrane treatment. The method comprises the steps of: smashing liquorice, and extracting with high temperature organic acid, ultrafiltration intercepting, concentrating and precipitating and separating by ethanol to obtain glycyrrhiza polysaccharide. The glycyrrhiza polysaccharide prepared by the invention is added to cigarettes by 0.05-0.2% of weight of cut tobacco, and the human bronchus epithelium BEAS-2B cell viability of a tobacco agglutinator of cigarettes obtained is remarkably higher than that of a blank sample, so that the glycyrrhiza polysaccharide has a positive effect.

The invention belongs to the technical field of molecular diagnosis, and particularly relates to IncRNA application, a kit and medicine. According to the application of IncRNA for preparing the kit for diagnosis and/or prognosis evaluation of NSCLC. The nucleotide sequence of the IncRNA is shown in SEQ ID No.1. The IncRNA is highly expressed in plasma before the NSCLC patient surgery, after the surgery, the expression level in the plasma is remarkably lowered, the expression levels in the NSCLC cancer tissue, the NSCLC patient plasma and the NSCLC cells are all remarkably higher than those ofpara-carcinoma tissue, healthy human plasma and normal bronchus epithelial cells. The study result suggests that the IncRNA has the remarkable capability for inhibiting the NSCLC cell tumor activity,and the IncRNA is newly found for tumor inhibition.

The present invention discloses a method for constructing a pulmonary nodule computer-aided detection model. The method comprises the following steps: obtaining bronchial epithelial cells of a subjectand extracting RNA; constructing a double-stranded cDNA library according to the RNA and conducting sequencing; comparing a sequencing result with a reference genome and selecting a differential significant gene and a differential significant variation of the subject; using a vector formed by combining the differential significant gene and the differential significant variation, and benign and malignant pulmonary nodules as sample data; randomly dividing the sample data of the subject into a training set and a prediction set and carrying out multiple times of training on the training set andthe prediction set on the basis of a support vector machine model adopting a radial basis kernel function to obtain a model output value of the prediction set; and carrying out non-dimensionalizationon clinical data and CT image data of the subject in the prediction set, deducing a Roc fitting curve by combining a non-dimensionalized data comprehensive value and the model output value, calculating a lower area, and adjusting covariance and penalty factors to be the value corresponding to the maximum value of the lower area to obtain the pulmonary nodule computer-aided detection model.

The invention provides a canine primary bronchial epithelial cell and an application thereof in preparation of immortalized cells, and belongs to the technical field of biology. The canine primary bronchial epithelial cell is prepared by taking 45-day-old clean male beagles; separating canine tracheal tissues, cutting into pieces, adding collagenase and trypsin to digest, so as to obtain intact bronchiole tissues; cultivating the obtained cells, and further carrying out indirect immunofluorescence assay to detect keratin 18 expression, wherein an experiment proves that the canine primary bronchial epithelial cell is successfully obtained. A eukaryotic expression vector pEGFP-hTERT containing telomerase genes (hTERT) is built; after the canine primary bronchial epithelial cell is transfected, the hTERT gene is highly expressed; the bronchial epithelial cell can be stably subcultured for at least 20 generations. According to the canine primary bronchial epithelial cell provided by the invention, important biological materials are provided for research of infection mechanism of canine pathogens such as canine influenza viruses.

The invention belongs to the field of tissue engineering and medical materials, and relates to a silk fibroin 3D printing based biological scaffold suitable for bronchial mucosa epithelial growth anda preparation method thereof. The biological scaffold is characterized in that silk fibroin/hydroxypropyl carboxymethyl cellulose is adopted as a scaffold material, a 3D printing and freeze drying combined technology is adopted, a bronchial epithelial cell line (BEAS-2B cells) is used as seed cells, the BEAS-2B cells grow on a porous 3D printed silk fibroin/hydroxypropyl carboxymethyl cellulose scaffold, and then the 3D printed silk fibroin/hydroxypropyl carboxymethyl cellulose scaffold is obtained. The silk fibroin 3D printing based scaffold can be used for tracheal defect repair and can provide meaningful reference for development of tissue engineering artificial trachea.

The invention discloses an application of 5-pentyl-3-methoxy-phenol to preparation of products for preventing and treating oxidative stress or inflammatory response induced diseases. The chemical formula of 5-pentyl-3-methoxy-phenol is shown in the description. 5-pentyl-3-methoxy-phenol is an Nrf2 (nuclear factor-erythroid 2-related factor 2) signaling pathway agonist and an NF (nuclear factor)-kappa B signaling pathway inhibitor. 5-pentyl-3-methoxy-phenol can up-regulate the protein levels of Nrf2 as well as phase II detoxification enzyme NQO1 and antioxidant enzyme GCLM which are regulated by Nrf2, increase the level of glutathione as an endogenous antioxidant and inhibit generation of exogenous toxicant induced reactive oxygen species, and has a protective effect on exogenous toxicant induced oxidative damage to pulmonary bronchial epithelial cells, glomerular mesangial cells, human nerve cells and human breast cancer cells. 5-pentyl-3-methoxy-phenol can inhibit activation of an NF-kappa B signaling pathway, down-regulate inflammatory gene expression and inhibit LPS induced inflammatory response of macrophage RAW 264.7 of a mouse.

The invention discloses a preparation method of a high-throughput airway inflammatory drug screening cell model based on an NF-kappaB signaling pathway, and an application of the cell model. The preparation method uses a bronchial smooth muscle cell or a bronchial epithelial cell as a source cell, NF-kappaB in the cell is activated by a stimulating factor, and the total NF-kappaB ratio and the phosphorylated NF-kappaB ratio in the cell are detected by a homogeneous time-resolved fluorescence technique to obtain the phosphorylation level of NF-kappaB; and parameters are adjusted to make a ratioof the phosphorylation level of NF-kappaB in a model group cell to the phosphorylation level of NF-kappaB in a normal group cell be equal to or more than 1.2 in order to obtain the cell model. The inflammatory cell model is constructed by screening the influences of the type and the density of the cell and the type, the concentration and the stimulation time of the stimulating factor on the phosphorylation of NF-kappaB and detecting the phosphorylation level of NF-kappaB in the cell through an HTRF technology, so a basis is provided for screening and studying of airway inflammation drugs.

Aspects of the present invention concern the identification of several methods to analyze the genes that are modulated in normal human bronchial epithelial (NHBE) cells after exposure to cigarette smoke condensates (CSC) or cigarette smoke (CS). Embodiments described herein include methods to identify a gene that is modulated in response to exposure to CSC or CS, methods to identify tobacco products that have a reduced potential to contribute to tobacco-related disease, methods to make tobacco products that have a reduced potential to contribute to a tobacco-related disease, methods to identify a subject's predilection to acquire a tobacco related disease, the use of particular genes as biomarkers for tobacco-related disease, and patterns of gene expression or genetic signatures that are unique to each particular tobacco product.

An object of the present invention is to provide a method capable of inexpensively and conveniently preparing cells having pluripotency and a very low risk of tumorigenic transformation. The cells having pluripotency and a very low risk of tumorigenic transformation can be prepared by suspension-culturing mammalian mesenchymal stem cells such as human mesenchymal stem cells from bone marrow (hMSC-BM) and human adipose tissue-derived mesenchymal stem cells (hAT-MSC) (also referred to as “human adipose-derived stem cells [hADSC]”), 7 types of human adherent mature cells (human hepatocyte cells [hHEP cells], human umbilical vein endothelial cells [HUVEC cells], human dermal lymphatic microvascular endothelial cells [HMVEC cells], human epidermal keratinocyte cells [NHEK cells], human bronchial epithelial cells [NHBE cells], human melanocyte cells [NHEM cells], and human smooth muscle cells [UASMC cells]), and 3 types of human adherent precursor cells (human dermal fibroblast cells [NHDF cells], human skeletal muscle myoblast cells [HSMM cells], and human osteoblast cells [NHOst cells]) to form cell masses (spheroids).

The invention discloses a construction method of an endothelial cell and pericyte co-culture model for researching tubulation. The method comprises the following steps: (1) digesting umbilical vein endothelial cells with trypsin to obtain umbilical vein endothelial single cells; (2) digesting pericytes into single cells of the pericytes by using trypsin, and then adding CFSE dye with the final concentration of 2-10 [mu]mol/L for staining to obtain stained pericytes; and (3) uniformly mixing the umbilical vein endothelial single cells and the stained pericytes according to the cell number ratio of (5-20):1, conducting culturing by using a bronchial epithelial cell culture medium, and observing the formation condition of the tube on a fluorescence microscope. According to the method disclosed by the invention, the influence of pericytes on the formation of endothelial cell tubes can be dynamically observed through a fluorescence microscope, the effect of human tumor pericytes on tumor blood vessels can be reflected through in-vitro experiments, and the method has good social and economic values.

The present invention relates to a composition for preventing or treating asthma, having, an active ingredient, a saturated or unsaturated fatty acid having 12 to 22 carbon atoms. The saturated or unsaturated fatty acid functions to inhibit or eliminate various asthma symptoms induced by ovalbumin. Specifically, the fatty acid inhibits the proliferation of leukocytes, neutrophils, lymphocytes and monocytes in BALF and serum, inhibits the proliferation of bronchial epithelial cells, reduces mucus in bronchioles, and reduces infiltration of inflammatory cells around bronchioles and blood vessels in a dose-dependent manner. Also, the fatty acid induces a decrease in the expression of IFN-γ, IL-12p40, IL-4, IL-5, IL-13 and TNF-α, which are Th2 cell-related cytokines. Therefore, the fatty acid overcomes the side effects of current asthma therapeutic agents, has an excellent therapeutic effect without having toxicity, and may advantageously be used as a composition for preventing, treating or alleviating asthma.

The invention discloses application of human GRK5 genes, namely application of human GRK5 genes serving as cancer cell specified action targets in preparation of drugs for treating non-small cell lungcancer. The cancer cell specified action targets are RNA interference effect targets. Compared with normal pulmonary bronchial epithelial cells, the GRK5 genes have high expressions in the non-smallcell lung cancer cells; RNA interfering lentivirus designed aiming at the GRK5 genes can inhibit GRK5 expression and is capable of obviously inhibiting proliferation of the non-small cell lung cancercells, blocking tumor cell division at a G2/M phase, promoting cells apoptosis, inhibiting cell migration and inhibiting formation of xenograft tumor in nude mice, and the aim of treating the non-small cell lung cancer can be further achieved. The human GRK5 genes can serve as treatment targets of the non-small cell lung cancer, and provide wide prospects for developing medicines in treatment of the non-small cell lung cancer based on the GRK5 genes in future.

The invention discloses the application of resveratrol in preparation of drugs for treatment of human bronchial epithelial cell injury. The resveratrol has a significant effect on ensuring the survival rate of human bronchial epithelial cells induced by divalent nickel ions, and can inhibit the cell damage induced by the divalent nickel ions by regulating the formation of reactive oxygen species clusters. The invention provides a new idea for the application of resveratrol in the prevention and treatment of the human bronchial epithelial cell injury induced by heavy metals.