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Chinese Journal of Biotechnology ; (12): 969-978, 2020.
Article in Chinese | WPRIM | ID: wpr-826879


Drugs targeting immune checkpoint are used for cancer treatment, but resistance to single drug may occur. Combination therapy blocking multiple checkpoints simultaneously can improve clinical outcome. Therefore, we designed a recombinant protein rPC to block multiple targets, which consists of extracellular domains of programmed cell death protein 1 (PD-1) and cytotoxic T lymphocyte-associated antigen 4 (CTLA-4). The coding sequence was inserted into expression vector and stably transfected into HEK293 cells. The culture supernatant was collected and rPC was affinity-purified. Real-time quantitative PCR was used to evaluate the expression levels of ligands for PD-1 and CTLA-4 in several human cancer cell lines. The binding of rPC with cancer cells was examined by immunofluorescence cell staining, the influence of rPC on cancer cell growth was assayed by CCK-8. The results showed that rPC could be expressed and secreted by stably transfected HEK293 cells, the purified rPC could bind to lung cancer NCI-H226 cells which have high levels of ligands for PD-1 and CTLA-4, no direct impact on cancer cell growth could be observed by rPC treatment. The recombinant protein rPC can be functionally assayed further for developing novel immunotherapeutic drugs for cancer.

Animals , CTLA-4 Antigen , Genetics , Cell Proliferation , HEK293 Cells , Humans , Lung Neoplasms , Metabolism , Programmed Cell Death 1 Receptor , Genetics , Protein Binding , Protein Domains , Genetics , Recombinant Fusion Proteins , Genetics , Metabolism
Protein & Cell ; (12): 213-224, 2012.
Article in English | WPRIM | ID: wpr-757278


The self-renewal and multipotent potentials in neural stem cells (NSCs) maintain the normal physiological functions of central nervous system (CNS). The abnormal differentiation of NSCs would lead to CNS disorders. However, the mechanisms of how NSCs differentiate into astrocytes, oligodendrocytes (OLs) and neurons are still unclear, which is mainly due to the complexity of differentiation processes and the limitation of the cell separation method. In this study, we modeled the dynamics of neural cell interactions in a systemic approach by mining the high-throughput genomic and proteomic data, and identified 8615 genes that are involved in various biological processes and functions with significant changes during the differentiation processes. A total of 1559 genes are specifically expressed in neural cells, in which 242 genes are NSC specific, 215 are astrocyte specific, 551 are OL specific, and 563 are neuron specific. In addition, we proposed 57 transcriptional regulators specifically expressed in NSCs may play essential roles in the development courses. These findings provide more comprehensive analysis for better understanding the endogenous mechanisms of NSC fate determination.

Animals , Astrocytes , Cell Biology , Metabolism , Cell Differentiation , Genetics , Gene Expression Profiling , Gene Regulatory Networks , Mice , Neural Stem Cells , Cell Biology , Metabolism , Oligodendroglia , Cell Biology , Metabolism , Protein Interaction Mapping
China Pharmacy ; (12)1991.
Article in Chinese | WPRIM | ID: wpr-518522


OBJECTIVE:To study the extraction process of ginsenoside.METHODS:The optimal extraction process was selected with the orthogonal design.The content of ginsenoside in ginseng was determined by UV-Spectrophotometry.RESUL_TS:The amount and concentration of alcohol and extraction time showed significant influence on extract obtained.CONCLUSI_ON:The optimal extraction process is as follows:adding 75% alcohol into crude ginseng(6∶1 in weight) and extracting 6 times for 45 min each extraction.