The tonic method is a commonly used treatment for tumors. In the clinic, there is a debate on whether it is beneficial or detrimental to treat tumors with this treatment method. Most scholars believe that tonic drugs can inhibit tumor growth, prolong the survival period of patients, and improve their quality of life [1]; while some clinicians believe that tonic drugs will promote tumor growth and should not be used more often in clinical practice. In order to explore the effect of tonic drugs on tumors, we chose the tonic representative formula Sijunzi Tang to observe its tumor suppression rate and induction of apoptosis of tumor cells in hormonal mice. 1. Materials and methods 1.1 Experimental animals 60 mice of Kunming breed, weighing 20g±2g, half male and half female, were purchased from the Laboratory Animal Center of the First Military Medical University. The mice were kept in separate cages, fed and watered freely, and the mouse room was kept ventilated. 1.2 Cell line The cell line was mouse SRS-82 ascites tumor strain, subcutaneous inoculation can generate sarcoma. The cell line is provided by the Institute of Pharmaceutical Research of this university, and is routinely resuscitated and cultured. 1.3 Drug preparation The drugs used in this experiment were purchased from the pharmacy of Nanfang Hospital, and the 5-FU injection was manufactured by Jiangsu Nantong Pharmaceutical General Factory (Su Wei Pharmaceutical Permit No. 241201). Sijunzi Tang composition: ginseng 9g, atractylodes macrocephala 9g, poria 9g, roasted licorice 6g. Preparation: conventional water decoction twice, the resulting decoction was evaporated and concentrated by a constant temperature water bath at 70 ℃, and the final concentration of Sijunzi Tang was obtained to be 0.33g/ml (containing raw drugs), which was stored in a refrigerator at 4 ℃. The 5-FU injection was diluted to 3mg/ml in sterile saline and stored in 4℃ refrigerator. 1.4 Establishment of tumor-bearing mouse model In vitro resuscitation culture of SRS-82 cells, collect the cells in the exponential growth phase of the cells, centrifuge at 1000r/min, wash twice with PBS, centrifuge to remove the supernatant, and dilute with sterile saline, adjusted to 2~3×106/ml. 10 healthy Kunming mice were randomly selected, and each was injected intraperitoneally with 0.4ml of the cell suspension mentioned above, and attention was paid to the observation of the inoculated Observe the growth of ascites in the inoculated mice. After about one week, the abdomen of the inoculated mice was obviously distended and protruded, and the ascites was extracted, which was milky white in color, and then diluted in sterile saline in a sterile test tube at a ratio of 1:2, and finally about 15 ml of ascites dilution solution was obtained. 36 Kunming mice were inoculated with the above ascites dilution, and each one was inoculated subcutaneously with 0.2 ml of the dilution in the right hind limb. 1.5 Experimental grouping and drug administration The 36 mice with successful modeling were randomly divided into 3 groups of 12 mice each and numbered within the groups.The 3 groups were: blank control group, Sijunzi Tang group, and 5-FU group. All treatments were started from the second day after modeling, the Sijunzi Tang group was gavaged with prepared Sijunzi Tang decoction, 0.5 ml each, once a day; the blank control group was gavaged with an equal amount of saline every day; and the 5-FU group was intraperitoneally injected with diluted 5-FU injection, 0.2 ml each, every other day. After the above treatments were carried out continuously for 15 days, the mice were killed and the tumors were stripped. 1.6 Weighing the tumor and calculating the tumor inhibition rate The tumor blocks were stripped, weighed on an electronic balance, and the tumor inhibition rate was calculated according to the conventional formula. 1.7 Transmission electron microscopy observation of tumor cell morphology and structure Two mice were randomly taken from each group for electron microscopy observation. After stripping the tumor body, 0.5mm3 volume of tumor tissue was cut and quickly fixed with glutaraldehyde. After dehydration, infiltration, embedding, sectioning, staining and other steps, the finished electron microscope sections were observed under transmission electron microscope. 1.8 Agarose gel electrophoresis After preparing the reagents, DNA was extracted and subjected to electrophoresis. 1% agarose electrophoresis, voltage 60V, electrophoresis time 1.5h. 1.9 Statistical treatment All the data were processed by SPSS10.0, and analyzed by analysis of variance (ANOVA) with random design. 2, Results 2.1 Tumor weight and tumor suppression rate The tumor weight and tumor suppression rate of each group are shown in Table 1. As shown in the table, the tumor weight of each administered group was significantly reduced compared with that of the blank control group, among which the tumor suppression effect of the 5-FU group was the most obvious, and the tumor weight was the lightest (P<0.01); Sijunzi Tang also had an obvious tumor-suppressing effect (P<0.05). Table 1 Mean tumor weight and tumor inhibition rate of each group Experimental group Tumor weight (g) (х±s) Tumor inhibition rate (%) Blank control group 4.78±1.53 Sijunzi Tang group 4.03±1.23* 15.69 5-FU group 3.12±1.08** 34.1 * P<0.05 vs. Blank group, ** P<0.01 vs. Blank group 2.2 Transmission electron microscopy observations The tumor weights of Sijunzi Tang and 5-FU groups were the most obvious (P<0.01). Most of the sections in the group and the 5-FU group showed changes of necrotic cells: pale staining of chromatin, swelling of organelles and vacuole-like changes. Both groups did not show a large number of cells undergoing apoptotic characteristic changes. 2.3 Agarose gel electrophoresis DNA ladder-like bands are the characteristic changes of cells undergoing apoptosis, and no obvious ladder-like bands of DNA electrophoresis were seen in both the Sijunzi Tang group and the 5-FU group. 3.Discussion The experimental results showed that the tumor weight of Sijunzi Tang group was significantly reduced compared with that of blank group, indicating that the formula has a positive effect on inhibiting the tumor size, and the clinical application of this formula is beneficial in the treatment of tumors. The morphology of apoptotic cells was characterized by compact nucleus and compressed cytoplasm at the initial stage, and then the nucleus fragmented and evolved into apoptotic vesicles, but other organelles remained intact. The most striking biochemical feature is the cleavage of DNA double bonds between nucleosomes to form 180-200 bp fragments that appear as "ladders" on DNA agarose gel electrophoresis [2]. In our experiments, through the observation under the electron microscope and the detection of agarose gel electrophoresis, the Sijunzi Tang group did not show obvious apoptotic signs, and the agarose electrophoresis did not show the typical DNA "trapezoidal band", so we did not see the typical apoptotic changes. This study showed that Sijunzi Tang showed obvious tumor suppression rate in tumor-bearing mice, and its mechanism was not related to the induction of apoptosis of tumor cells. Sijunzi Tang is mainly tonic and may not have the effect of directly attacking the evil. In the in vivo experiments, the Sijunzi Tang group showed significant tumor suppression, which may act through overall other pathways. Literature reports that tonic drugs have positive effects on immune function and survival time of hormonal animals in in vivo experiments [3]. Sijunzi Tang can significantly increase the activity of NK cells in hormonal mice, and the anti-tumor mechanism of Sijunzi Tang-like tonic drugs may mainly improve the immune function of the body, enhance the activity of NK cells and LAK cells in the body, and play an indirect killing role [3], which can help to dispel the evil by supporting the positive. The experimental results suggest that Sijunzi Tang, which belongs to tonic benefit, can significantly inhibit tumor growth in hormonal mice, which proves that this kind of traditional Chinese medicine has obvious tumor-suppressing effect in vivo. Sijunzi Tang did not induce apoptosis of tumor cells in vivo, and its effect on tumors may be achieved through other actions.