Potential Mechanism of Chinese herbal formula Sijunzi Decoction in Treating Gastric Cancer
On the terminal day (Day 21 post-MFC inoculation), all mice were euthanized. Gastric carcinoma tumor tissues from the MFC allograft were immediately dissected, snap-frozen in liquid nitrogen, and stored at -80°C until nucleic acid extraction to preserve RNA integrity. Four-to-five-week-old inbred 615 mice were maintained under specific pathogen-free (SPF) conditions at 22 °C ± 2 °C, 55% ± 5% humidity, with a 12/12-hour light/dark cycle, and provided with ad libitum access to water and diet. A one-week acclimatization period was provided prior to any experimental procedures. After acclimatization, mice were randomly divided into three groups (n=6/group):  Model group: Received 200μl of 0.9% saline via intragastric gavage on day 1, followed by MFC cell inoculation on day 7.  Early SJZD group (E-SJZD): Received 200μl of Sijunzi Decoction (19,200 mg/kg/bw) via intragastric gavage on day 1, followed by MFC cell inoculation on day 7.  Synchronization SJZD group (S-SJZD): Received 200μl of Sijunzi Decoction (19,200 mg/kg/bw) and MFC cell inoculation simultaneously on day 7. All groups received daily intragastric administration for 3 consecutive weeks. RNA from snap-frozen tissues samples was extracted using TRIzol (Invitrogen, USA) according to manufacturer's instructions. mRNA was isolated from total RNA extracts. Strand-specific RNA-seq libraries were constructed from the isolated mRNA. 