Keywords：方药研究, 实验研究, 配方资产, 转化沟通, 2.5 胸腺、脾脏指数测定
Section Index
1 Experimental Materials
1.1 Experimental Drugs
Pei's Hematopoietic Granules (composed of Rehmannia root, Chinese yam, Moutan bark, Prince ginseng, Northern sand ginseng, Codonopsis pilosula, Cinnamon twig, Jujube, Schisandra berry, honey-fried licorice, White peony root, etc., with each packet containing 36.25g of crude herbs). Specification: 18g per packet, produced by the Gansu Provincial Academy of Medical Sciences, batch number: 080310.
Zhenqi Fuzheng Granules, specification: 15g per bag, manufactured by Hunan Shidai Sunshine Pharmaceutical Co., Ltd., batch number: National Drug Approval No. Z43021035.
1.2 Experimental Animals and Tumor Strains
Sixty healthy SPF mice, half male and half female, weighing 20±2g, aged 6–8 weeks, provided by the Experimental Animal Center of Gansu Provincial College of Traditional Chinese Medicine.
Research on Pei Zhengxue's series of formulas
Provided, animal license number: SCXK (Gan) 2004-0006. H22 (liver cancer) tumor strain, provided by the Pharmacology and Toxicology Research Center of the Gansu Provincial Academy of Medical Sciences.
1.3 Experimental Reagents
- KH₂PO₄: Beijing Hongxing Chemical Factory
- Anhydrous ethanol: Tianjin Bodi Chemical Co., Ltd.
- NaOH: Tianjin Chemical Reagents No. 3 Factory
- KCl: Tianjin Chemical Reagents No. 5 Factory
- Na₂HPO₄·12H₂O: Tianjin Kemio Chemical Reagents Development Center
- NaCl: Beijing Chemical Factory
- Propidium iodide (PI): Sigma Company
- Formaldehyde: Xi'an Chemical Reagents Factory
- H₂O₂ (analytical grade): Shanghai Wusong Chemical Fertilizer Factory, China
- Xylene (analytical grade): Tianjin Baishi Chemical Co., Ltd.
Citrate buffer solution: Add 3g of trisodium citrate (C₆H₅Na₃O₇·2H₂O) and 0.4g of citric acid (C₆H₈O₇·H₂O) to 1000ml of distilled water.
- Trisodium citrate (C₆H₅Na₃O₇·2H₂O): Xi'an Chemical Reagents Factory
- Citric acid (C₆H₈O₇·H₂O): Xi'an Chemical Reagents Factory
Phosphate-buffered saline (PBS): Add 8.00g of NaCl, 3.48g of Na₂HPO₄·12H₂O, 0.20g of KCl, and 0.20g of KH₂PO₄ to 1000ml of distilled water, adjust the pH to 7.4, sterilize at 120℃ for 20 minutes, and store at 4℃ for later use.
- Rabbit anti-mouse VEGF monoclonal antibody: Zhongshan Jinqiao Bioengineering Co., Ltd.
- Horseradish peroxidase-labeled goat anti-mouse IgG (secondary antibody): Zhongshan Jinqiao Bioengineering Co., Ltd.
- Horseradish peroxidase-labeled streptavidin working solution: Zhongshan Jinqiao Bioengineering Co., Ltd.
- Bovine serum albumin: Zhongshan Jinqiao Bioengineering Co., Ltd.
- Normal goat serum blocking solution: Zhongshan Jinqiao Bioengineering Co., Ltd.
- DAB chromogenic kit: Zhongshan Jinqiao Bioengineering Co., Ltd.
- Mouse IL-12 ELISA kit: Zhongshan Jinqiao Bioengineering Co., Ltd.
1.4 Experimental Instruments and Equipment
| Instrument Name | Model/Origin |
|---|---|
| Binocular biological microscope | OLYMPUS CHC-212 |
| CO₂ incubator | SHEL-LAB 1825TC, USA |
| 1/1000 electronic balance | JA-2003, Shanghai |
| Rapid mixing vortex mixer | SK-1, Shenzhen |
| Refrigerator | Jinwangzi, Beijing |
| Electric heating drying oven | 202-2, Shanghai |
| Biological purification workbench | SW-CJ-1F, Suzhou |
| Electric heating constant temperature water bath | S-648, Shanghai |
| Centrifuge | LD4-2A, Beijing |
| Pressure cooker | BOX, Shanghai |
| Sterile operating box | Type B-small, Shanghai |
| Water bath constant temperature incubator | PYX-DHS-50×60, Shanghai |
| Large-capacity desktop freezer centrifuge | TDL5M, Changsha High-tech Industrial Development Zone |
| Micropipette | WKY type 5-25μl, Shanghai |
2 Experimental Methods
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