1 Experimental Materials

1 Experimental Materials

1.1 Experimental Drugs

Peishishengxue Granule (composed of Shengdi, Shanyao, Danpi, Taizishen, Beishashen, Dangshen, Guizhi, Dazao, Wuweizi, Zhigancao, Baishao, etc., manufactured by the Gansu Provincial Academy of Medical Sciences as a granular preparation, with each packet containing 36.25 g of crude herbs), Zhenqi Fuzheng Granule (provided by Foci Pharmaceutical Co., Ltd., with national drug approval number Z62020416).

1.2 Experimental Animals and Tumor Strains

Sixty healthy BALB/C mice, half male and half female, weighing 20 ± 2 g and aged 5–6 weeks, provided by the Experimental Animal Center of the Gansu Provincial Academy of Medical Sciences, with animal qualification certificate number: Yi Dong Zi No. 04-007. 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

• NaCl: Beijing Chemical Plant
• Na₂HPO₄·12H₂O: Tianjin Kemiou Chemical Reagent Development Center
• NaOH: Tianjin Chemical Reagent Factory No. 3
• KCl: Tianjin Chemical Reagent Factory No. 5
• Anhydrous Ethanol: Tianjin Bodi Chemical Co., Ltd.
• KH₂PO₄: Beijing Hongxing Chemical Plant
• Isopropyl Alcohol: Shanghai Jianxin Chemical Co., Ltd.
• Dimethyl Sulfoxide (DMSO): Tianjin Dengfeng Chemical Reagent Factory
• Formaldehyde: Xi'an Chemical Reagent Factory
• Acetone: Chengdu Chemical Reagent Factory
• Concanavalin A (ConA): Beijing Dingguo Biotechnology Development Center

Phosphate Buffered Saline (PBS): 8.0 g NaCl, 3.48 g Na₂HPO₄·12H₂O, 0.20 g KCl, 0.20 g KH₂PO₄, mixed with triple-distilled water to make 1000 ml, adjusted to pH 7.4, sterilized at 121°C for 20 minutes, and stored at 4°C for later use.

Lymphocyte Separation Solution: Tianjin Biotechnology Development Center

RPMI1640 Powder: Product of Gibco Company in the United States. When used, take 10.4 g of dry culture powder and 2.0 g of NaHCO₃ powder, dissolve them in 1000 ml of triple-distilled water, stir until completely dissolved, then adjust the pH to 7.2–7.4 using 1 mol/L HCl, filter through a sterile filter to remove bacteria, aliquot, and store at 4°C for later use.

Research on Pei Zhengxue's Series of Formulas

Fetal Bovine Serum: Product of Hangzhou Sijiqing Bioengineering Materials Co., Ltd., inactivated by 56°C water bath for 30 minutes.

MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide): Product of Sigma Company. Dissolve MTT in PBS (pH 7.4) to prepare a 5 mg/ml solution, store at 4°C away from light.

Mouse IL-2 ELISA Kit: Product of Beijing Jingmei Bioengineering Co., Ltd.

Total RNA Extraction Kit: Product of Shanghai Bioengineering Technical Service Co., Ltd.

RT-PCR Reaction Kit: Product of Shanghai Bioengineering Technical Service Co., Ltd.

Diethyl Pyrocarbonate (DEPC): Product of Sigma Company in the United States.

DNA Marker: Product of Shanghai Bioengineering Technical Service Co., Ltd.

Ethidium Bromide (EB): Product of Shanghai Bioengineering Technical Service Co., Ltd.

Agarose: Product of Huamei Bioengineering Company.

β-actin primers, IL-2 primers, IFN-γ primers: Synthesized by Shanghai Bioengineering Technical Service Co., Ltd.

0.25% Bromophenol Blue: Product of Shanghai Bioengineering Technical Service Co., Ltd.

10×TBE Electrophoresis Buffer: Tris 108 g, Boric Acid 55 g, EDTA (0.5 mol/L, pH 8.0) 40 ml.

1.4 Experimental Instruments and Equipment

2 Experimental Methods