Keywords：方药研究, 实验研究, 配方资产, 转化沟通, 3.3 裴氏升血颗粒对小鼠H₂₂肿瘤组织中突变型P53表达的影响
Section Index
1 Experimental Materials
1.1 Experimental Drugs
Peishishengxue Granule (composed of Rehmannia, Chinese yam, Moutan bark, Prince ginseng, Radix Adenophorae, Codonopsis pilosula, Cinnamon twig, Jujube, Schisandra, Honey-fried licorice, White peony, etc., manufactured by the Gansu Provincial Academy of Medical Sciences as a granular oral solution, containing 36.25g of crude herbs per packet).
Zhenqi Fuzheng Granule (provided by Foci Pharmaceutical Co., Ltd., with national drug approval number Z62020416).
1.2 Experimental Animals and Tumor Strains
Sixty healthy Kunming mice, half male and half female, weighing 20±2g, aged 5–6 weeks, provided by the Experimental Animal Center of the Gansu Provincial College of Traditional Chinese Medicine, with animal qualification certificate number: Yi Dong Zi (06) No. 004. The H22 (liver cancer) tumor strain was provided by the Pharmacology and Toxicology Research Center of the Gansu Provincial Academy of Medical Sciences.
1.3 Experimental Reagents
| Reagent | Source |
|---|---|
| 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 |
| rh-Annexin-V-FITC/PI double-stained flow cytometry reagent kit | Bender, Hangzhou Lianke agent |
| Dimethyl sulfoxide (DMSO) | Tianjin Dengfeng Chemical Reagents Factory |
Phosphate-buffered saline (PBS): 8.00g NaCl, 3.48g Na2HPO4·12H2O, 0.20g KCl, 0.20g KH2PO4, diluted with triple-distilled water to 1000ml, adjusted to pH 7.4, autoclaved at 120℃ for 20 minutes, stored at 4℃ for later use.
| Reagent | Source |
|---|---|
| Rabbit anti-human P53 monoclonal antibody | Beijing Zhongshan Jinqiao Biotechnology Co., Ltd. |
| Immunohistochemistry SP reagent kit (anti-rabbit) | Beijing Zhongshan Jinqiao Biotechnology Co., Ltd. |
| DAB chromogenic reagent kit | Beijing Zhongshan Jinqiao Biotechnology Co., Ltd. |
| APES anti-peeling reagent | Beijing Zhongshan Jinqiao Biotechnology Co., Ltd. |
| PBS buffer | Beijing Zhongshan Jinqiao Biotechnology Co., Ltd. |
1.4 Experimental Instruments and Equipment
| Instrument/Equipment | Model/Origin |
|---|---|
| Flow cytometer (FCM) | Coulter Company, USA |
| Inverted microscope | OLYMPUS PM-6, Japan |
| Binocular biological microscope | OLYMPUS CHC-212 |
| CO2 incubator | SHEL-LAB 1825TC, USA |
Research on Pei Zhengxue’s series of formulas and medicines
1 Experimental Instruments
- 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
- West cold refrigerator: BY-160, Hangzhou
- Pressure cooker: BOX, Shanghai
- Sterile operating box: Type B-small, Shanghai
- Water bath constant temperature incubator: PYX-DHS-50×60, Shanghai
- Large-capacity desktop frozen centrifuge: TDL5M, Changsha High-tech Industrial Development Zone
- Spectrophotometer: VIS-7220, Beijing
- Electrophoresis tank: DYCP31A type, Beijing
- Electrophoresis apparatus: HV-3000 type, Beijing
- PCR amplification instrument: PE2400 type, American PE company
- Gel imaging analysis system: BTS-20.M type, Uvitec company
- Micropipette: WKY type 5-25μl, Shanghai
2 Experimental Methods
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