47 results about "Soft agar" patented technology

A therapeutic composition that includes an active portion of the lysyl oxidase pro-peptide in a pharmaceutically acceptable carrier substance and methods of using such a therapeutic composition are disclosed. The active agent does not have lysyl oxidase enzymatic activity. Preferably, the active polypeptide is active in inhibiting cell growth in soft agar and active in inhibiting tumor formation. In addition, the active polypeptide preferably comprises an active portion of the amino acid sequence given in SEQ ID NO.: 1 or SEQ ID NO.: 2, or conservative substitions thereof. Alternatively, the active polypeptide comprises a polypeptide comprising an active portion of an amino acid sequence selected from the group consisting of SEQ ID NOs.: 3-8, or conservative substitions thereof.

The present invention relates to methods and compositions designed for the treatment, management, or prevention of cancer, particularly, metastatic cancer. In one embodiment, the methods of the invention comprise the administration of an effective amount of an antibody that binds to EphA2 and agonizes EphA2, thereby increasing EphA2 phosphorylation and decreasing EphA2 levels. In other embodiments, the methods of the invention comprise the administration of an effective amount of an antibody that binds to EphA2 and inhibits cancer cell colony formation in soft agar, inhibits tubular network formation in three-dimensional basement membrane or extracellular matrix preparation, preferentially binds to an EphA2 epitope that is exposed on cancer cells but not non-cancer cells, and / or has a low Koff, thereby, inhibiting tumor cell growth and / or metastasis. The invention also provides pharmaceutical compositions comprising one or more EphA2 antibodies of the invention either alone or in combination with one or more other agents useful for cancer therapy.

The present invention relates to methods and compositions designed for the treatment, management, or prevention of cancer, particularly, metastatic cancer. In one embodiment, the methods of the invention comprise the administration of an effective amount of an antibody that binds to EphA2 and agonizes EphA2, thereby increasing EphA2 phosphorylation and decreasing EphA2 levels. In other embodiments, the methods of the invention comprise the administration of an effective amount of an antibody that binds to EphA2 and inhibits cancer cell colony formation in soft agar, inhibits tubular network formation in three-dimensional basement membrane or extracellular matrix preparation, preferentially binds to an EphA2 epitope that is exposed on cancer cells but not non-cancer cells, and / or has a low Koff, thereby, inhibiting tumor cell growth and / or metastasis. The invention also provides pharmaceutical compositions comprising one or more EphA2 antibodies of the invention either alone or in combination with one or more other agents useful for cancer therapy.

The invention relates to a method for preparing jarosite through microbial conversion. The method comprises the following steps: (1) preparing a 9K culture medium; (2) preparing a microbial conversion culture medium; (3) preparing a soft agarose culture medium; (4) performing culture preservation, namely mixing leptospirillum ferriphilum BYQ bacteria liquid in a stable phase preserved at the temperature of 0 DEG C with the soft agarose culture medium, solidifying at room temperature, and preserving at the temperature of 2-4 DEG C; (5) activating the strain, namely inoculating the leptospirillum ferriphilum BYQ strain into a shake flask containing the 9K culture medium and performing shaking culture, thus obtaining a strain activation solution; (6) performing biological conversion, namely inoculating the strain activation solution to a shake flask containing the microbial conversion culture medium and performing shaking culture, and standing at room temperature, thus obtaining jarosite which is attached to the wall of the shake flask and is generated through biological conversion; (7) soaking and centrifuging the shake flask to which the jarosite is attached by using a hydrochloric acid solution, thus obtaining a precipitate; and (8) performing vacuum drying on the precipitate to constant weight, and grinding to obtain the jarosite. The method is rapid, simple, convenient and environmentally friendly.

It is intended to provide a detection and analysis system for cancer cell colonies, whereby an environmental cell carcinogen causing cell carcinogenesis or a chemical or a food inhibiting cell carcinogenesis can be quickly and accurately detected, and a method therefor. Namely, a detection and analysis system for cancer cell colonies using a culture system (1) prepared by the agar overlaying method which comprises a bottom layer (1) being composed of a culture medium, soft agar and a carcinogenesis inducer and / or an anticancer agent and having a definite size and a top layer (12) being composed of a culture medium, soft agar and cells, and further provided with an optical microscope (21), an electronic data conversion unit (22) such as a digital camera and a computer system (23) for processing the data converted by the electronic data conversion unit (22). This computer system (23) has an image analysis software stored therein whereby the electronic data are grayed, calibrated and binarized by subtraction and single threshold and thus the presence or absence of colonies, the number of colonies, the size distribution of colonies, etc. are analyzed.

The Rev peptide that binds to nucleophosmin / B23 with the highest affinity exhibits the greatest cytotoxicity on Ras-3T3 cells and inhibits tumor growth most effectively in nude mice. The efficiency of colony formation in soft agar of Ras-3T3 cells is significantly inhibited by treatment with Rev peptide. In addition, Rev peptide can potentiate the doxorubicin-induced decrease of cellular viability in U1 bladder cancer cells and inhibition of tumor growth in nude mice. Treatment of Rev peptide increases protein expression and transcriptional activity of p53 and inhibits the nucleophosmin / B23-mediated PCNA promoter activation. Peptides having high affinity of binding to molecular targets such as nucleophosmin / B23 represent a useful approach to anti-cancer biotherapeutics.

This invention provides a quantitative assay technique for soft agar colony formation that can be carried out rapidly with high accuracy. Specifically, this invention relates to a method for evaluation of cell survival comprising: a step of overlaying agar at the bottom of a vessel with agar containing cells, overlaying the agar containing cells with a medium and culturing the cells; a step of removing the medium, adding tetrazolium that produces water-soluble formazan and an electron carrier and culturing the cells; and a step of evaluating cell survival based on the color developed by the water-soluble formazan.

The invention relates to an identification method for salmonella paratyphi B and salmonella paratyphi B var Java in salmonella species, and mainly solves the technical problems of rapid screening and identification of salmonella paratyphi B from clinical diarrhea cases and environment and food samples. According to the technical scheme, a kit for identifying salmonella paratyphi B and salmonella paratyphi B var Java is adopted. The kit is characterized by comprising the following raw materials: (1) a salmonella B group immune rabbit anti-serum which contains diluted serums of salmonella O:4 and O:5 factor antibodies; (2) a salmonella H-phase immune rabbit anti-serum which contains diluted serums of salmonella H:b and H:2 factor antibodies; (3) a potassium tartrate biochemical identification tube which contains 10.0 g of peptone, 10.0 g of sodium potassium tartrate, 5.0 g of sodium chloride, 0.024 g of phenol red, 15.0 g of agar and 1000.0 ml of distilled water; (4) sterilizing mineral oil; (5) H-phase dedicated inducing soft agar which contains 10.0 g of tryptic soy agar, 5.0 g of casein, 1.0 g of potassium nitrate, 2.0 g of agar powder and 1000.0 ml of distilled water; (6) an H:b factor inducing serum which contains inducing serum H:b factors.

The present invention provides a human cervical adenocarcinoma cell line, which is characterized by naming as a primary cervical adenocarcinoma drug-resistant strain W038, and is deposited in the China Center for Type Culture Collection with a preservation number being CCTCC No: C2017246. The primary cervical adenocarcinoma drug-resistant strain W038 has weak invasion ability and weak migration ability, and can be used as a double-negative control group for invasion and migration. The in-vitro soft agar colony formation test proves that the tumorigenicity proves that the cloning ability is strong, and the in-vitro susceptibility test that the strain is resistant to a variety of drugs, which is consistent with clinical features. The preparation method disclosed by the invention can purify cancer cell lines to over 99%, wherein the screened medium component can selectively kill histocyte, has high selectivity for tumor cell growth, and can increase the proliferation rate of tumor cells and shorten the culture period. The method can be used not only to purify the cervical adenocarcinoma lines, but also to purify and prepare other cancer cell lines.

Targeting metabolic enzymes in human cancer Abstract Lung cancer is a devastating disease and a major therapeutic burden with poor prognosis. The functional heterogeneity of lung cancer (different tumor formation ability in bulk of tumor) is highly related with clinical chemoresistance and relapse. Here we find that, glycine dehydrogenase (GLDC), one of the metabolic enzyme involved in glycine metabolism, is overexpressed in various subtypes of human lung cancer and possibly several other types of cancers. GLDC was found to be highly expressed in tumor-initiating subpopulation of human lung cancer cells compared with non-tumorigenic subpopulation. By array studies we showed that normal lung cells express low levels of GLDC compared to xenograft and primary tumor. Functional studies showed that RNAi inhibition of GLDC inhibits significantly the clonal growth of tumor-initiating cells in vitro and tumor formation in immunodeficient mice. Overexpression of GLDC in non-tumorigenic subpopulation convert the cells to become tumorigenic. Furthermore, over-expression of GLDC in NIH / 3T3 cells and human primary lung fibroblasts can transform these cells, displaying anchorage-independent growth in soft agar and tumor-forming in mice. Not only is GLDC is expressed human lung cancer, it is also up-regulated in other types of cancer, such as colon cancer. RNAi knockdown of GLDC in colon cancer cell line, CACO-2 cells, can also inhibit the tumor formation in mice. Thus GLDC maybe a new metabolic target for treatment of lung cancer, and other cancers.

The invention discloses a preparation method of an immortalized human intervertebral disc nucleus pulposus cell system. The preparation method is as follows: extrinsic human telomerase reverse transcriptase is transfected into the human nucleus pulposus cell to activate telomerase and construct immortalized human nucleus pulposus cell line through induction immortalized human nucleus pulposus cell line, normal cells not transformed cells are used, wherein the cells have normal growth rate and karyotype, have contact inhibition, anchorage dependence and other safety properties and do not have carcinogenicity and soft agar cloning capability; and as same as the germline stem cells, the cells have real immortalization, thus the standard cell line can be provided for the further research of the pathogenic mechanism of the degenerative disc disease and a new idea is provided to increase the entire control level of the degenerative disc disease.

The invention discloses a hair follicle microarray co-culture system and application in treating pathological alopecia. The hair follicle microarray co-culture system comprises a hair follicle stem cell suspension and damaged hair follicles to be repaired. According to the invention, a new idea for creating a microenvironment suitable for hair follicle repair by combining allogeneic normal human hair follicle stem cells with drug molecules is provided, and the hair follicle microarray co-culture system for performing in-vitro 3D co-culture on hair follicle stem cells and damaged hair folliclesby using soft agar is provided.

The present invention provides a human cervical adenocarcinoma cell line, which is characterized by naming as a primary cervical adenocarcinoma drug-resistant strain W038, and is deposited in the China Center for Type Culture Collection with a preservation number being CCTCC No: C2017246. The primary cervical adenocarcinoma drug-resistant strain W038 has weak invasion ability and weak migration ability, and can be used as a double-negative control group for invasion and migration. The in-vitro soft agar colony formation test proves that the tumorigenicity proves that the cloning ability is strong, and the in-vitro susceptibility test that the strain is resistant to a variety of drugs, which is consistent with clinical features. The preparation method disclosed by the invention can purify cancer cell lines to over 99%, wherein the screened medium component can selectively kill histocyte, has high selectivity for tumor cell growth, and can increase the proliferation rate of tumor cells and shorten the culture period. The method can be used not only to purify the cervical adenocarcinoma lines, but also to purify and prepare other cancer cell lines.