82 results about "Cell signaling pathways" patented technology

This invention is directed to a method for preparation of a biological sample for measurement of protein epitopes that allows for the preservation of intracellular protein epitopes and detection of signal transduction pathways based on the ability to capture transient activation states of the epitopes. The method provided by the invention allows for the rapid fixation of biological samples containing red blood cells, to ensure that epitopes of signal transduction molecules and other intracellular protein epitopes are preserved in the active state. The method of the invention further allows for lysis of red blood cells, thereby making it a useful method for cytometric analysis of biological samples, including, for example, whole blood, bone marrow aspirates, peritoneal fluids, and other red blood cell containing samples. The invention also provides a method to recover or “unmask” epitopes on intracellular antigens that have been made inaccessible by the cross linking fixative necessary to fix the sample. Significantly, the methods of the invention allow preservation and analysis of phospho-epitope levels in biological samples taken directly from patients to determine disease-specific characteristics.

An embodiment of the present invention is useful for identifying tumor cells in bladder cancer washes, pleural effusions, biliary tumor cells and circulating tumor cells in whole blood. Subsequent analysis may identify therapeutic treatments based on a single cell analysis of activatable elements in cell signaling pathways. This analysis can be useful for diagnosis, prognosis, therapy selection and monitoring of solid tumor diseases.

This invention is directed to a method for preparation of a biological sample for measurement of protein epitopes that allows for the preservation of intracellular protein epitopes and detection of signal transduction pathways based on the ability to capture transient activation states of the epitopes. The method provided by the invention allows for the rapid fixation of biological samples containing red blood cells, to ensure that epitopes of signal transduction molecules and other intracellular protein epitopes are preserved in the active state. The method of the invention further allows for lysis of red blood cells, thereby making it a useful method for cytometric analysis of biological samples, including, for example, whole blood, bone marrow aspirates, peritoneal fluids, and other red blood cell containing samples. The invention also provides a method to recover or ''unmask'' epitopes on intracellular antigens that have been made inaccessible by the cross linking fixative necessary to fix the sample. Significantly, the methods of the invention allow preservation and analysis of phospho-epitope levels in biological samples taken directly from patients to determine disease-specific characteristics.

The present invention belongs to the technical field of cell biology and particularly relates to a technology for regulating and controlling a Jak-Stat pathway to differentiate, dedifferentiate and rejuvenate cells, and an application of the technology. Rejuvenated cell products and / or cell products of different lineage types are obtained by small molecule compound combination, cytokine combination or recombinant protein combination, a gene editing technology and a transgenic technology to quantitatively and / or regularly regulate and control gene or protein targets of the Jak-Stat signaling pathways in cells. The provided technology for regulating and controlling the Jak-Stat signal pathway, and the product / derivatives of the cell product can be used for re-programming cells, tissues, organs and organisms, construct tissue engineering materials, repair damages of tissues and organs of mammals and the aged and degenerated tissues and organs, delay or reverse aging processes of the cells, tissues, organs and organisms, and can be used for immunoregulation of the cells, tissues, organs and organisms.

The present invention relates to a method for activating natural killer cells (NK cells), and more particularly, to a method for enhancing the cytotoxicity of natural killer cells by inducing the overexpression of suppressor of cytokine signaling 2 (SOCS2) which is a protein involved in cell-signaling pathways in natural killer cells. The inventors of the present invention observed that when natural killer cells were treated with IL-15, a cytokine involved in natural killer cell differentiation, the expression of SOCS2 increased and the expression of proline-rich tyrosine kinase 2 (Pyk2) was inhibited by the SOCS2, the expression of which increased. In addition, when Pyk2 was overexpressed, the ability to produce IFN-γ and the ability to kill tumor cells of natural killer cells decreased. Therefore, SOCS2 can be used for activating natural killer cells and the natural killer cells activated by the method can be used for the prevention or treatment of cancer.

This invention is directed to a method for preparation of a biological sample for measurement of protein epitopes that allows for the preservation of intracellular protein epitopes and detection of signal transduction pathways based on the ability to capture transient activation states of the epitopes. The method provided by the invention allows for the rapid fixation of biological samples containing red blood cells, to ensure that epitopes of signal transduction molecules and other intracellular protein epitopes are preserved in the active state. The method of the invention further allows for lysis of red blood cells, thereby making it a useful method for cytometric analysis of biological samples, including, for example, whole blood, bone marrow aspirates, peritoneal fluids, and other red blood cell containing samples. The invention also provides a method to recover or “unmask” epitopes on intracellular antigens that have been made inaccessible by the cross linking fixative necessary to fix the sample. Significantly, the methods of the invention allow preservation and analysis of phospho-epitope levels in biological samples taken directly from patients to determine disease-specific characteristics.

The invention belongs to the technical field of biomedicines and in particular relates to a construction method and application of a double-target-spot DNA (Deoxyribonucleic Acid) nano therapy system.The construction method comprises the following steps: dissolving six single-stranded linear-chain DNA into an in-vitro buffer salt solution; carrying out annealing and self-assembling to form a DNAtetrahedron solution; adding crosslinking agent Sulfo-SMCC powder into a 5'-end C6 amino modified ssDNA solution; reacting at room temperature for 2h; removing an unreacted crosslinking agent and thenadding polypeptide powder; reacting at the room temperature for 12h and removing excessive polypeptide to obtain ssDNA-peptide; adding the ssDNA-peptide into the DNA tetrahedron solution, so as to finish construction of the double-target-spot DNA nano therapy system. According to the construction method provided by the invention, a constructed DNA tetrahedron carrier has a stable structure; aftera medicine is loaded on the carrier, the stability of a polypeptide medicine can be improved; the double-target-spot DNA nano therapy system has relatively strong targeting performance and mainly express a biological therapy effect through regulating and controlling a cell signal path.

The invention provides a method for inducing formation of porcine fat cells by a cell signal channel inhibitor. The method comprises the following steps of uniformly inoculating porcine embryo fibroblasts into a Corning cell culture plate according to a ratio of 4-6* 104 cells / cm2; culturing by using an inducing culture medium, and observing the cells, and performing oil red O dyeing to prove that the cells are fat cells after culturing for 20 days in a culture box containing 5% CO2 at 37 DEG C. The cell signal channel inhibitor instead of a transgene is used for inducing cell transformation,so that genic mutation caused by insertion of an exogenous gene into a cell genome can be effectively prevented; by the method, the research on the forming mechanism of the fat cells is facilitated to a certain extent; and the method with convenience and safety can be widely applied and can be used for researching a fat forming mechanism and also can play a roll in disease therapy and other clinical applications.

The invention provides methods of using CARD4 molecules to screen for modulators of LPS-induced cell signaling pathways. Also included are CARD-4 deficient mice, methods of modulating LPS-induced cell signaling pathways, and methods of treating or preventing bacterial infections.

The invention discloses a protective effect of a combination of gastrodin and rhynchophylla to human vein vascular endothelial cells. Through adding homocysteine with a certain concentration to induce and damage the endothelial cells, a hypertension cell damage model is built. By using the model, combination drugs of gastrodin and rhynchophylla with various concentrations are added in the model, the biochemical indexes such as lactic dehydrogenase (LDH), malondialdehyde (MDA) and nitric oxide (NO) are detected, so as to test the protective effect of the combination drug to the damaged endothelial cells. The expression of key protein on a latent signal channel is observed by the Western Blotting technology, the observation proves that the latent channel is the cell signal channel for the combination drug to generate the protective effect. In the invention, the effective constituents in two natural blood-pressure reducing drugs of gastrodin and rhynchophylla are creatively combined, and the protective effect of the combination drug to the hypertension cell damage model is researched. The effect of the combination is more remarkable than that of any single component, and the mechanism is clear, therefore the combination drug can become the antihypertensive drug of secondary development of the traditional Chinese medicine.

A non-invasive method for determining the presence or severity of a bodily disorder associated with hyperactivity or increased expression of a signal transduction protein, transcription factor, or protein kinase that is a member of the MAPK or GPCR pathways. A reagent that binds to such a signal transduction protein, transcription factor, or protein kinase is non-invasively contacted to a tissue or fluid within the body of a subject or to a fluid removed from a subject, the reagent is permitted to bind to the signal transduction protein, transcription factor, or protein kinase in the tissue or fluid, the presence of binding of the reagent to the signal transduction protein, transcription factor, or protein kinase in said tissue or fluid is determined, and the binding is correlated with the presence or severity of said bodily disorder within the subject.

The invention discloses a cell signal pathway detection kit and a detection method based on a mass spectrum flow technology. Through the kit and a matched signal pathway stimulant and a detection antibody, and by utilizing a mass spectrum flow detection technology, an intracellular signal path activation level can be specifically and effectively detected at a high throughput and a single cell level, the sample detection precision is greatly improved, and a false negative analysis result possibly existing in a traditional detection means is avoided to a great extent.

The invention provides a microfluidic system and method to rapidly analyze large numbers of compounds or complex mixtures of compounds, particularly, low abundance cellular proteins involved in cell signaling pathways. In one aspect, an integrated microfluidic system comprises an upstream separation module (preferably, a multi-dimensional separation device), a microfluidic device for on-device protein digestion of substantially separated proteins received from the upstream separation module, a downstream separation module for separating digestion products of said proteins, a peptide analysis module and a processor for determining the amino acid sequence of said proteins. Preferably, the system comprises an interfacing microfluidic device between the downstream separation module and the peptide analysis module.

The present invention relates to compositions and methods associated with cocktails of growth factors and small molecules that target specific cell signaling pathways, the cocktails having been formulated to allow / promote the expansion of progenitor cells (e.g., nephron progenitor cells) within a defined culture system.

The invention discloses a method for isolated culture of esophagus epithelial stem cells. The method comprises the steps of taking esophagus mucosa tissue and epithelium adhering to the esophagus mucosa tissue, removing muscular tissue, taking a basal layer of epithelium mucosae tissue, shearing the basal layer of epithelium mucosae tissue into bits, conducting digestion centrifugation, and spreading obtained cells in a culture flask, culture plate or culture dish coated with a matrix; then adding a DMEM / Ham's F-12 culture solution containing HEPES, sodium bicarbonate, L-glutamic acid, FBS, EGF, insulin, hydrocortisone, vibrio cholerae toxin, transferring, BPE, quasi-vitamin A and cell signaling pathway inhibitor; finally, placing the culture flask, culture plate or culture dish containing cell suspension in a 37 DEG C incubator for culture, and obtaining the single-layer epithelial stem cells in 10-14 days. According to the method, operation is convenient, safety performance is high, the esophagus epithelial stem cells are obtained through separation, the activity of the stem cells is high, and application prospects in tissue engineering and regenerative medicine are broad.

The invention discloses a humanized antibody and application thereof, the humanized antibody comprises a heavy chain variable region and a light chain variable region, and can specifically target a CH1 structural domain 162 site sialylated epitope of IgG, the amino acid sequence of the heavy chain variable region is as shown in SEQ ID NO. 1, and the amino acid sequence of the light chain variable region is as shown in SEQ ID NO. 2. The humanized antibody can effectively inhibit the growth, invasion and suspension growth ability of tumor cells, has an obvious anti-tumor effect, and also can inhibit the activation of a c-Met / Wnt signal channel and an FAK signal channel of the tumor cells.

The invention belongs to the technical field of culture media, and particularly relates to a reprogramming medium for a human induced pluripotent stem cell (hiPSC). The reprogramming medium comprisesan adult cell culture medium for culturing a human somatic cell and a cell signaling pathway inhibitor added to the adult cell culture medium; and the cell signaling pathway inhibitor is any one of ora combination containing more of a glycogen synthase kinase 3 inhibitor, a mitogen-activated protein kinase kinase inhibitor and a transforming growth factor beta inhibitor. The reprogramming mediumis obtained through adding one or more small-molecular compounds to adult cell culture medium, is used for induced reprogramming culture or induced differentiation culture, is a non-trophoblastic animal origin-free medium, greatly improves the speed and the efficiency of reprogramming of the human somatic cell into the human pluripotent stem cell, and especially can efficiently reprogram the humansomatic cell into the hiPSC for expressing specific markers.

The invention discloses a protein sensitizer of tumor chemotherapeutics and an application thereof, the protein sensitizer comprises Survivin mutant protein, the protein sensitizer has 90% of homology with Survivin through gene engineering preparation, the Survivin comprises one point mutation, multipoint mutation, combinatorial mutation and simultaneous mutation of sites of 20 site, 34 site, 84 site and 117 site. The protein sensitizer is combined with tumor chemotherapeutics for usage, the tumor chemotherapeutics refers to one or more of doxorubicin, epirubicin, daunorubicin or paclitaxel. According to the effect of Survivin protein in cell signal pathway and the influence of Survivin protein on multi-drug resistance reversing, the chemotherapeutics is combined with TmSm, the TmSm is capable of effectively promoting the killing effect of the chemical medicines, effectively reducing the application amount of the chemical medicines and increasing the killing effect of the chemical medicines on cells, and is the effective protein sensitizer of tumor chemotherapeutics.

The present invention discloses a method for modifying ubiquitin and inhibiting an ubiquitination pathway. The modification is achieved by a family of effector proteins secreted by a type III secretion system in Gram-negative bacteria. The family of the effector proteins is a type of adenosine diphosphate (ADP) ribosyl transferase, specifically performs single ADP ribosylation modification on threonine residue at a 66th locus of a ubiquitin molecule in eukaryotes, and also is free of modification on ubiquitin-like proteins SUMO and NEDD8. The modification of the ubiquitin cannot be reversiblyde-modified by hydrolytic enzymes capable of removing ADP ribosylation in eukaryotic cells, and thus effectively destroys ubiquitin-mediated signaling pathways in host cells. The method plays important functions on killing tumor cells, changing cell signaling pathways, etc.

The present invention relates to a method comprising inferring activity of a PI3K cellular signaling pathway in a tissue and / or cells and / or a body fluid of a medical subject based at least on expression levels of one or more target gene(s) of the PI3K cellular signaling pathway measured in an extracted sample of the tissue and / or the cells and / or the body fluid of the medical subject. The present invention further relates to an apparatus comprising a digital processor configured to perform such a method, a non-transitory storage medium storing instructions that are executable by a digital processing device to perform such a method, and a computer program comprising program code means for causing a digital processing device to perform such a method.

The invention, which belongs to the technical field of biological medicine, relates to drug combination for treating non-small cell lung cancer, in particular to application of combination of ergosterol and gefitinib. Lung cancer A549 cells and PC-9 cells are cultured in vitro to research the inhibition effect of the medicine on tumors. With two system cell lines (respectively PC-9 cells sensitiveto GEF and drug-resistant A549 cells), an MTT testing is performed; a Hochest 33258 cell apoptosis test is performed; cell apoptosis and cell cycle are detected by using flow cytometry; and the influence of the EGFR signal path of the cell is examined by a Western-blots test. Whether ERG and GEF combined application has a synergistic lung cancer resisting effect or not, whether the synergistic and sensitizing effects can be realized for sensitive cell strains or not and whether the sensitivity of drug-resistant cell strains can be improved or not are preliminarily discussed; and the treatmenteffect and mechanism research on non-small cell lung cancer are also discussed.

The invention discloses an application of bone morphogenetic protein-4 in screening drugs for resisting cardiac hypertrophy, heart failure or cardiac fibrosis and belongs to the field of biomedicine. Screened drug types comprise: bone morphogenetic protein-4 formation inhibitor, bone morphogenetic protein-4 antagonistic, bone morphogenetic protein-4 receptor antagonists and bone morphogenetic protein-4 downstream signal antagonistic. The invention also relates to an application of the drugs in preparation of medicines for resisting cardiac hypertrophy, heart failure and arrhythmia. Researches have found that bone morphogenetic protein-4 can induce myocardial hypertrophy, apoptosis and cardiac fibroblast collagen secretion increase, lead to occurrence of cardiac hypertrophy, heart failure and arrhythmia. However, drugs for antagonism of bone morphogenetic protein-4 and its cell signaling pathway can be used to inhibit occurrence of cardiac hypertrophy, inhibit cardiac fibrosis and minimize occurrence of cardiac cell death and arrhythmia. The invention provides a theoretical basis for realizing high-flux medicine screening, and is of practical guiding significance.

There is provided a cell line that is prepared by transforming HEK293 cell with a human Kir2.1 gene using a retrovirus expression system, wherein the HEK293 expresses a stable α1G T-type calcium channel. The cell line responds sensitively to KCl and forms an appropriate level of the membrane voltage so that the cell signaling pathway may be investigated by the molecular biological and biochemical studies.

There is provided a cell line that is prepared by transforming HEK293 cell with a human Kir2.1 gene using a retrovirus expression system, wherein the HEK293 expresses a stable α1G T-type calcium channel. The cell line responds sensitively to KCl and forms an appropriate level of the membrane voltage so that the cell signaling pathway may be investigated by the molecular biological and biochemical studies.

The invention provides a method for regulating a cellular pathway by using plant hormone ABA (Abscisic Acid) and a small molecular substance PYR (Pyrabactin). The method comprises the following steps:constructing ABA receptor proteins ABI1 and PYR1 in Arabidopsis thaliana on a human cell expression vector, then transferring into HEK293T cells respectively, adding ABA in order that the two receptor proteins undergo induced interaction, and adding a PYR inhibitor in order that the interaction degree is weakened; fusing other human cell signal pathway key proteins with the two receptor proteins,and making the two receptor proteins undergo interaction under the induction of the ABA, so that a fused signal pathway also undergoes ABA induced aggregation. According to the method provided by theinvention, a key protein LRP6 on a human Wnt pathway is selected. After fusion expression of an ABA receptor and LRP6, LRP6 aggregates under the induction of ABA, thereby promoting the production ofbeta-catenin and opening up an intracellular signal pathway. After the PYR inhibitor is added, the production amount of the beta-catenin is reduced, and the cellular pathway is closed.

The invention provides a subarachnoid hemorrhage prediction model establishment method and system based on gene and cell signal pathways. The method comprises the steps: obtaining normal brain cell group 1 and SAH brain cell gene chip data, and preprocessing; carrying out differential expression analysis and signal path analysis on the data of the normal brain cell group 1 and the SAH brain cell gene chip; carrying out LCN2 intervention on a normal brain cell group 2 and SAH brain cells to obtain RNA-Seq data under different conditions, and preprocessing to form LCN2 data; carrying out differential expression analysis and signal path analysis on the LCN2 data; carrying out feature selection on the differentially expressed gene data obtained through the differential expression analysis to obtain feature gene data; and obtaining a training sample based on the feature gene data, training a plurality of classifiers based on the training sample, integrating the trained classifiers, and establishing a prediction model. According to the method, the subarachnoid hemorrhage model can be accurately established, and the treatment target can be obtained.

The invention belongs to the technical field of cosmetics, in particular to novel embedded type eight-component medicinal powder with the whitening and freckle removing functions and a preparation method thereof. The novel embedded type eight-component medicinal powder is mainly prepared from the following components in parts by mass: 10 parts of white muscardine silkworms, 10 parts of tribulus terrestris, 10 parts of bletilla striata, 10 parts of white poria, 10 parts of white mulberry root-bark, 10 parts of radix ampelopsis, 10 parts of white peony roots and 10 parts of pearl powder. According to the novel embedded type eight-component medicinal powder and the preparation method thereof, the activity of tyrosinase is regulated and controlled in a synergistic manner according to differentactivation cell signaling pathways of the components, and in addition, the medicinal powder can be fully embedded in liposome and play a role in activity and is an organic combination of a traditional active substance and a modern preparation technology; and the defects that traditional eight-component medicinal powder has a certain stimulation effect and can cause anaphylactic reaction can be overcome, and a good whitening effect is achieved.

The present invention discloses an anti-tumor polypeptide and an application thereof in preparing anti-tumor drugs. Aiming at a molecular structure, a ligand binding mode and amino acid sequence characteristics of a NRP-1 transmembrane region, and the transmembrane region capable of specifically binding the human source NRP-1 molecular structure and binding ligands is designed to block binding ofNRP-1 with a variety of the ligands of VEGF, EGF, and HGF, thereby blocking the anti-tumor polypeptide drugs of downstream cellular signaling pathways. The anti-tumor polypeptide binds to the NRP-1 transmembrane region monomer competitively, interferes dimerization of NRP-1, inhibits the binding of the NRP-1 to the ligands, thus inhibits activation of various tumor cell signaling pathways, and achieves purposes of inhibiting tumor cell proliferation, migration and infiltration, and promoting apoptosis. The provided anti-tumor polypeptide provides a new technical means for treatment of tumors.

The invention discloses a GLP-1 receptor agonist, and belongs to the field of biological medicine. according to the invention, a high-throughput drug screening technology is applied to screen the polypeptide named 100-44 (PL00201001) from a large cyclic peptide library, and the polypeptide can activate a GLP-1 receptor and cause activation of a cell signal channel. PL00201001 contains 80 amino acid cyclic peptides, a series of derived small cyclic peptides and linear peptides are obtained through a decompression technology, and both the small cyclic peptides and the linear peptides have the effect of a GLP-1 receptor agonist. The PL00201001 and a series of polypeptides thereof can be used as GLP-1 receptor agonists, and new polypeptide drugs for diabetes, obesity and related diseases thereof can be further developed.

The invention discloses an anti-tumor polypeptide with double effects and an application thereof. The anti-tumor polypeptide with double effects has an amino acid sequence shown as SEQ ID NO.4. The polypeptide provided by the invention can specifically target NRP-1 molecules expressed on endothelial cells and tumor cells, improve the permeability of vascular endothelial cells and increase the permeability of tumor tissues, thereby improving the capacity of a drug penetrating through a vascular barrier and permeating into the tumor tissues. Meanwhile, proliferation of cancer cells can be inhibited and apoptosis of the cancer cells can be promoted by blocking a downstream cell signal path regulated and controlled by NRP-1. The polypeptide disclosed by the invention can be used for treating various solid tumors including pancreatic cancer, and can improve the effect of treating malignant tumors by using drugs, especially using a monoclonal antibody targeted drug.