1 Experimental Materials

1 Experimental Materials

1.1 Experimental Drugs

Pei's Soft Liver and Anti-Bloating Pill (PRGXP) (composed of salvia miltiorrhiza, astragalus, bupleurum, paeonia lactiflora, curcuma wenyujin, seaweed, soapberry thorns, hedyotis diffusa, and polygonum cuspidatum), specification 6 g per pack, Gansu Provincial Academy of Medical Sciences, production batch number: 090210.

Compound Cantharidin Capsules, specification 0.25 g per capsule, Shaanxi Fangzhou Pharmaceutical Co., Ltd., production batch number: 090401.

1.2 Experimental Animals and Tumor Strains

Seventy-two healthy SPF-grade Kunming mice, half male and half female, weighing 20 ± 2 g. All experimental mice were provided by the Experimental Animal Center of Gansu University of Traditional Chinese Medicine, with animal qualification certificate number: Medical Animal No. SCXK (Gansu) 2004-0006.

H22 (liver cancer) tumor strain: introduced from the Beijing Institute of Materia Medica of the Chinese Academy of Medical Sciences and propagated and preserved by the Pharmacology and Toxicology Research Center of the Gansu Provincial Academy of Medical Sciences.

1.3 Experimental Reagents

• Rabbit anti-mouse P27 monoclonal antibody: Wuhan Bode Bioengineering Co., Ltd.
• Rabbit anti-mouse Bcl-2 monoclonal antibody: Wuhan Bode Bioengineering Co., Ltd.
• DAB chromogenic reagent kit: Wuhan Bode Bioengineering Co., Ltd.
• BSA blocking solution: Wuhan Bode Bioengineering Co., Ltd.
• SABC: Wuhan Bode Bioengineering Co., Ltd.
• Anhydrous ethanol: Tianjin Bodi Chemical Co., Ltd.
• NaOH: Tianjin Chemical Reagents Factory No. 3
• Na₂HPO₄·12H₂O: Tianjin Kemiou Chemical Reagents Development Center
• NaCl: Beijing Chemical Factory
• KH₂PO₄: Beijing Red Star Chemical Factory
• KCl: Tianjin Chemical Reagents Factory No. 5
• H₂O₂ (analytical grade): Shanghai Wusong Fertilizer Factory, China
• Propidium iodide (PI): Sigma Company
• Formaldehyde: Xi'an Chemical Reagents Factory
• Xylene (analytical grade): Tianjin Baishi Chemical Co., Ltd.

Citrate buffer: Add 3 g of trisodium citrate (C₆H₅Na₃O₇·2H₂O) and 0.4 g of citric acid (C₆H₈O₇·H₂O) to 1000 ml of distilled water, both from Xi'an Chemical Reagents Factory.

Phosphate buffer (PBS): Add 8 g of NaCl, 3.48 g of Na₂HPO₄·12H₂O, 0.2 g of KCl, and 0.2 g of KH₂PO₄ to 1000 ml of distilled water, adjust pH to 7.4, sterilize at 120°C for 20 minutes, and store at 4°C for later use.

1.4 Experimental Instruments