32 results about "Proliferation assay" patented technology

This invention provides an agonist antibody to a human thrombopoietin receptor (alias: human c-Mpl). More particularly, this invention provides an agonist antibody to a human thrombopoietin receptor, wherein the agonist antibody comprises: antibody constant regions comprising (1) amino acid sequences in a heavy chain constant region and a light chain constant region of a human antibody, (2) an amino acid sequence of a heavy chain constant region with a domain substituted between human antibody subclasses, and an amino acid sequence of a light chain constant region of a human antibody, or (3) amino acid sequences comprising a deletion(s), substitution(s), addition(s), or insertion(s) of one or several amino acid residues in the amino acid sequences of (1) or (2) above; and antibody variable regions capable of binding to and activating a human thrombopoietin receptor; and wherein the agonist antibody has the properties: (a) that the antibody induces colony formation at a concentration of 10,000 ng / ml or lower as determined by the CFU-MK colony formation assay using human umbilical-cord-blood-derived CD34+ cells; and (b) that the antibody has a maximal activity at least 50% higher than that of PEG-rHuMGDF and an 50% effective concentration (EC50) of 100 nM or less in the cell proliferation assay using UT7 / TPO cell. Also provided is a pharmaceutical composition for treating thrombocytopenia comprising said antibody.

The invention relates to the technical field of medicines, in particular to a lipoic acid modified intrinsically disordered protein nano-carrier, a preparation method thereof and an application of the nano-carrier. The nano-carrier is crosslinked by disulfide bonds of lipoic acid, formed polypeptide polymer can be rapidly degraded in cells and cannot be accumulated in the cells, and amino acid forming a polypeptide carrier exists in vivo and has no toxic and side effects on the cells and a human body. CCK-8 cell proliferation assay indicates that the prepared nano-carrier has low cell toxicity and good gene and chemotherapeutic drug co-carrier capacity, can successfully deliver sensitivity of autophagic specificity enhanced cancer cells to chemotherapeutic drugs caused by siRNA suppressive chemotherapeutic drugs in breast cancer therapy, promotes breast cancer cell apoptosis and accordingly becomes a targeted, efficient and low-toxicity nano-scale delivery system in breast cancer therapy.

The present invention provides an antiangiogenic polypeptide having the amino acid sequence set forth in SEQ ID NO: 1, or a fragment thereof, which is effective to inhibit endothelial cell proliferation as determined by the capillary EC proliferation assay. Preferably, the fragment has at least 50% inhibition of bFGF-stimulated EC proliferation at 5 μg / ml-20 μg / ml, more preferably 75% inhibition, most preferably 95% inhibition.

The present inventors obtained, from a phage library of human antibodies, an anti-mouse NR 10 neutralizing antibody-expressing BM095 clone that shows a strong proliferation-suppressing activity in an IL-31-dependent Ba / F3 cell proliferation assay system. When this anti-mouse NR 10 neutralizing antibody was administered to NC / Nga mice, a model of atopic dermatitis which is a mouse model of chronic dermatitis that arises as a result of repeated applications of picryl chloride, a mouse model of rheumatoid arthritis, and a mouse model of osteoarthritis, a significant effect of symptom suppression was observed. This revealed that the anti-NR 10 neutralizing antibody is indeed effective as a therapeutic agent for inflammatory diseases. In addition, the present inventors successfully obtained an anti-human NR 10 neutralizing antibody, providing extremely useful therapeutic agents with practical clinical applications.

The present invention provides an antiangiogenic polypeptide having the amino acid sequence set forth in SEQ ID NO: 1, or a fragment thereof, which is effective to inhibit endothelial cell proliferation as determined by the capillary EC proliferation assay. Preferably, the fragment has at least 50% inhibition of bFGF-stimulated EC proliferation at 5 μg / ml-20 μg / ml, more preferably 75% inhibition, most preferably 95% inhibition.

The invention relates to a novel application of fusion protein TAT-DCF1. By judging dcf1-induced U251 cell line apoptosis by virtue of such methods as immuno-electron microscope, atomic force microscope, CCK8 proliferation assay, JC-1 dyeing, nude mouse tumor heterotopic transplantation and the like, results show that dcf1, which is positioned in mitochondria, can cause pathological change of the mitochondria structure and result in drop in membrane potential, so that expression amounts of related genes are changed, and the apoptosis is finally induced. Nude mouse experiments show that them dcf1 can obviously diminish tumor volume of glioma, so that tumor growth is inhibited.

The invention discloses a method for inhibiting gastric cancer angiogenesis by using micro vesicles as miRNA (Micro Ribonucleic Acid) transport carriers. Through extraction and identification of micro vesicles, quantitative analysis of miRNA, protein extraction, Western blot, ELISA (Enzyme-linked Immuno Sorbent Assay) and EdU (5-Ethynyl-2'-deoxyuridine) proliferation assays, an in-vitro angiogenesis assay, a tumorigenesis assay and a tail intravenous injection micro vesicle assay, the situation that the quantitative analysis of the content of corresponding miRNA in artificially modified micro vesicles shows that the content of miR-29 is obviously increased by being compared with a negative control is discovered, the VEGF (Vascular Endothelial Growth Factor) protein secretion level of gastric cancer cells is obviously inhibited after the gastric cancer cells are hatched by the micro vesicles containing the miR-29, the proliferation of vascular endothelial cells co-cultured by the gastric cancer cells treated by the micro vesicles secreted by HEK (Human Embryonic Kidney)-293 cells treated by high-expression miR-29a and miR-29c is obviously inhibited, ring formation of the vascular endothelial cells co-cultured by the gastric cancer cells treated by the micro vesicles secreted by the HEK-293 cells treated by the high-expression miR-29a and the miR-29c is obviously inhibited, and the tumor volume in a mouse containing the micro vesicles which are subjected to the tail intravenous injection and high-expression miR-29a / c treatment is obviously smaller than the tumor volume in a mouse treated by the corresponding negative control.

Methods for enhancing survival and / or proliferation of neural stem cells and pharmaceutical compositions containing neural stem cells prepared by such methods, together with methods for assaying factors enhancing survival and / or proliferation of neural stem cells and methods for screening for such factors.Either Galectin-1 is overexpressed in neural stem cells or neural stem cells are cultured in a liquid medium containing Galectin-1. Pharmaceutical compositions containing Galectin-1-overexpressing neural stem cells and pharmaceutical composition containing Galectin-1, prepared by the aforementioned methods, improve higher cerebral functions damaged by cerebral ischemia. Further, by seeding neural stem cells at clonal concentrations and determining whether the seeded neural stem cells are capable of proliferating in an assay medium to be assayed, whether the factor enhances survival and / or proliferation of neural stem cells is assayed and a factor enhancing survival and / or proliferation of neural stem cells are identified using this assay method.

Protein kinase C plays important roles in TNBC development and could be a specific target. The in vitro anti-proliferative activity of PKC inhibitor chelerythrine on a panel of breast cancer cell lines was evaluated. Chelerythrine selectively inhibited the growth of TNBC cell lines over non-TNBC cell lines as demonstrated by cell proliferation assay and colony formation assay. The selective anti-proliferative effect of chelerythrine was associated with the differential apoptosis induction ability on breast cancer cell lines and induction of cell cycle arrest in some TNBC cell lines. By analysis of the expression levels of different PKC subtypes, the inventors found multiple PKC isozymes may mediate the selective activity of chelerythrine on TNBC cells. Finally, combination of chelerythrine and chemotherapy reagent taxol showed synergistic / additive effect on TNBC cell lines.

The invention discloses a nucleoside and base hydroxamic acid compound with DNA transmethylase and / or histone deacetylase inhibition activity, and a preparation method and applications thereof. The structural formula of the nucleoside and base hydroxamic acid compound is shown as the formula I, wherein R is nucleoside and base, nucleoside and base with substituent group or nucleoside and base analogue in DNA and / or RNA; Linker is a connecting chain connecting R with hydroxamic acid function group, the connecting chain comprises but is not limited within alkyl chain, alkyl chain with heteroatom, alkyl chain with aromatic ring, and alkyl chain with heterocycle. In-vitro cell proliferation assay show that the compound shown in the formula I can well inhibit the proliferation of leukemia cell K562 and histocyte lymphoma cell U937. Transmethylase and histone deacetylase inhibition assay shows that the compound shown in the formula I is a compound having inhibition activity on the DNA transmethylase and / or histone deacetylase. The formula I is as shown in the specification.

The embodiments herein disclose a method for synthesizing antagonistic peptide VEGF and bFGF. The method comprises synthesizing the antagonistic peptide for VEGF and bFGF and analyzing the purity of peptides. The quality of antagonistic peptide for VEGF and bFGF is analyzed by HPLC chromatogram and Mass spectrometry analysis. The biochemical activity of the antagonistic peptide for VEGF and bFGF is analyzed by competitive binding assay, cell proliferation assay, Matrigel assay for anti-angiogenic activity analysis, histopathological staining and Western blot analysis. The competitive binding assay of antagonistic peptide for VEGF and bFGF illustrate that peptides binds with cell receptors at a concentration of 2000 ng / ml. The cell proliferation assay illustrates that cell growth is arrested when antagonistic peptide for VEGF and bFGF are at a concentration of 2000 ng / ml. The anti-angiogenic activity analysis illustrates that angiogenesis is arrested when the concentration of antagonistic peptide for VEGF and bFGF is 2000 ng / ml.

The invention relates to the technical field of medicines, in particular to a lipoic acid modified intrinsically disordered protein nano-carrier, a preparation method thereof and an application of the nano-carrier. The nano-carrier is crosslinked by disulfide bonds of lipoic acid, formed polypeptide polymer can be rapidly degraded in cells and cannot be accumulated in the cells, and amino acid forming a polypeptide carrier exists in vivo and has no toxic and side effects on the cells and a human body. CCK-8 cell proliferation assay indicates that the prepared nano-carrier has low cell toxicity and good gene and chemotherapeutic drug co-carrier capacity, can successfully deliver sensitivity of autophagic specificity enhanced cancer cells to chemotherapeutic drugs caused by siRNA suppressive chemotherapeutic drugs in breast cancer therapy, promotes breast cancer cell apoptosis and accordingly becomes a targeted, efficient and low-toxicity nano-scale delivery system in breast cancer therapy.

Methods for producing stimulated, positive and negative control reference standard for monitoring intracellular cytokine levels and cytokine release in test samples by stimulating cells to produce cytokines in the presence of a cytokine release inhibitor, fixing the stimulated cells with a fixative such as paraformaldehyde, washing to remove excess fixatives and freeze-drying the stimulated, fixed cells. Methods for producing labeled reference standards for cell proliferation assays are also disclosed, in which proliferation-competent mammalian cells, isolated from a human or animal body are labeled with a label, such as a dye, that is divided between daughter cells during cell proliferation (e.g., carboxyfluorescein succinimidyl ester), the cells are stimulated to proliferate, the proliferated cells are fixed by addition of a fixative and then preserved by freeze drying or cryopreservation.