New medicinal application of interleukin-12
A technology of interleukin and application, which is applied in the direction of drug combination, extracellular fluid disease, medical preparations containing active ingredients, etc., can solve problems such as death, infection, and low immune function of patients, and achieve the purpose of broadening the scope of use and benefiting effect of effect
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Embodiment 1
[0023] Example 1 Mouse Bone Marrow Mononuclear Cells (HSC) IL-12R Flow Cytometry Analysis
[0024] 1 Reagents and instruments
[0025] Reagents: RPMI-1640 (GIBCO, USA), fetal bovine serum (GIBCO, USA), Ficoll lymphocyte separation medium (Shanghai Huajing), CD3-PE, CD4-PreCP, CD8-FITC, IFN-γ-APC, Rat IgG1-APC and BFA were purchased from BD Pharmingen (Becton Dickinson / Phar Mingen MSA).
[0026] Instruments: flow cytometer (FACS Calibur, BD Company, the United States), SW-CJ-2F clean bench (Suzhou Antai Air Technology Co., Ltd.), carbon dioxide incubator (SANYO, Japan), LXJ-II centrifuge ( Shanghai Medical Analytical Instrument Factory), XW-80A Vortex Mixer (Shanghai Jingke Industrial Co., Ltd.).
[0027] 2 Experimental method:
[0028] (1) Preparation of bone marrow nucleated cells
[0029] Eight BALB / c mice were sacrificed by neck dislocation. The femurs on both sides were taken and put into the complete RPMI1640 culture medium supplemented with 10% fetal bovine serum. Th...
Embodiment 2
[0043] Example 2 Expression of HSC Apoptotic Rate in Mouse Bone Marrow Nucleated Cells After Radiation Irradiation
[0044] Purpose of the experiment: To observe the preventive and therapeutic effects of IL-12 on radiation-damaged cells
[0045] 1.1 Experimental method
[0046] SPF grade Kunming rats were selected and irradiated with 4Gy by a linear accelerator. The mice were administered once 24 hours before and 24 hours after radiation. They were killed 4 days after irradiation, and the apoptosis rate of bone marrow HSCs was detected. HSCs were labeled with c-kit APC and AnnexinVPE. It is an indicator of apoptosis detection.
[0047] (1) Preparation of bone marrow nucleated cells
[0048] BALB / c mice were sacrificed by neck dislocation, and bilateral femurs were taken under aseptic conditions, cultured with RPMI164 complete culture medium (with 10% fetal bovine serum added), and the bone marrow was obtained by repeatedly blowing and blowing with a syringe, and separated wi...
Embodiment 4
[0102] Example 4 The effect of rhIL-12 on the repair of radiation-damaged hematopoietic stem cells in neonatal umbilical cord blood
[0103] 1 Experimental materials
[0104] 1.1 Reagents
[0105] RPMI-1640 medium, Hank's solution, and fetal bovine serum were all products of GIBCO Company in the United States, Ficoll lymphocyte separation medium was purchased from Shanghai Huajing Biotechnology Company, Annexin V apoptosis signal detection kit, and anti-human CD34-FITC were purchased from BD Pharmingen Company (Becton Dickinson / Phar Mingen USA), RPMI-1640 complete culture solution is RPMI-1640 culture solution containing 10% fetal bovine serum.
[0106] 1.2 Instruments
[0107] Flow cytometer (FACS Calibur, BD company, the United States), SW-CJ-2F clean bench (Suzhou Antai Air Technology Co., Ltd.), carbon dioxide incubator (SANYO, Japan), LXJ-II centrifuge (Shanghai Medical Analytical Instrument Factory), XW-80A vortex mixer (Shanghai Jingke Industrial Co., Ltd.), linear a...
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