| Objective: Studies showed that intestinal flora is involved in the occurrence and development of colorectal cancer.Fusobacterium nucleatum(F.nucleatum)is an opportunistic pathogen.In recent years,a large number of researches have found that F.nucleatum could cause colorectal cancer,compromise therapeutic efficacy due to chemotherapy resistance.It’s worth noting that Sijunzi Decoction has a greater advantage in regulating the intestinal flora.Because of this,this study compared the abundance and diversity of intestinal flora in the traditional Chinese medicine combined chemotherapy group,the chemotherapy group,the colon cancer group,and the healthy group by high-throughput sequencing technology.Real-time fluorescent quantitative PCR was used to determine the abundance of F.nucleatum and explore the effect of Sijunzi Decoction combined with chemotherapy on the abundance of F.nucleatum and the richness and diversity of intestinal flora in patients with colorectal cancer.Meanwhile,the occurrence of toxic and side effects of chemotherapy and the changes of symptoms of spleen deficiency in patients with colorectal cancer was also observed,to evaluate the effect of Sijunzi Decoction in the treatment of colorectal cancer.Methods: In this study,feces of patients with spleen deficiency syndrome of colorectal cancer and healthy people were collected that met the criteria for entry.Group A contains25 healthy people who passed the physical examination.Colorectal cancer patients(spleen deficiency)were divided into three groups,B,C,and D.Group B contains 25 patients with colorectal cancer who was first-diagnosed,30 patients with colorectal cancer who had got chemotherapy in group C,and 30 patients with colorectal cancer who had got traditional Chinese medicine combined with chemotherapy in group D.Extracted the total bacterial DNA in each sample,designed primers based on the 16 Sr DNA V3-V4 region for amplification,used the Illumina Miseq platform for high-throughput sequencing,and analyzed the sequencing results through the biological information analysis software to obtain the information of the sample.Analyzed and compared the differences in the abundance and diversity of the four groups of intestinal flora.In addition,real-time fluorescent quantitative PCR technology is used to obtain the abundance of F.nucleatum in the four groups,and the abundance of F.nucleatum in the D Chinese medicine combined chemotherapy group is compared with the other three groups.Meanwhile,the changes of symptoms of spleen deficiency before and after treatment,and the side effects of chemotherapy during treatment were observed to explore the effect of Chinese medicine compound Sijunzi Decoction combined with chemotherapy on the intestinal flora of patients with colon cancer and the abundance of F.nucleatum to evaluate the effect of Sijunzi Decoction in the treatment of colorectal cancer.Results:1.The results of Alpha diversity analysis indicated that the four groups were compared in pairs.The Chao index and Ace index of the A healthy group and the B colon cancer group were statistically significant(P<0.01).The A healthy group was compared with the C chemotherapy group,It was statistically significant(P<0.05),and the richness of the intestinal flora in the healthy group was higher than the cancer group and the chemotherapy group.The comparison between the D Chinese medicine combined chemotherapy group and the B colon cancer group was statistically significant(P<0.01).The D Chinese medicine combined chemotherapy group and the C chemotherapy group were statistically significant(P<0.05).The flora of the Chinese medicine combined chemotherapy group,The abundance is higher than the colorectal cancer group and the chemotherapy group,indicating that the Sijunzi Decoction combined with chemotherapy can significantly increase the intestinal flora richness of patients with colorectal cancer.Although the Shannon index and Simpson index of the four groups of samples were not statistically significant(P>0.05),indicating that there was no significant difference in the community diversity between the groups,the intestinal flora diversity of the Chinese medicine combined chemo-therapy group was similar to that of healthy people,indicating The combination of Sijunzi Decoction and chemotherapy can normalize the diversity of intestinal flora in patients with colorectal cancer.2.The results of Beta diversity analysis indicated: the difference between the four groups of A healthy group,B colon cancer group,C chemotherapy group,and D Chinese medicine combined chemotherapy group is greater than the difference within the group,but the difference is not significant(P>0.05).The number of cases in the group is less related.3.The real-time fluorescent quantitative PCR results showed that the abundance of F.nucleatum in the four groups in descending order was: B colon cancer group(5.81 ±0.29),C chemotherapy group(5.45 ±0.51),A healthy group(5.05 ±0.60),D Chinese medicine combined with chemotherapy group(4.59 ±0.28).After pairwise comparison,the comparison between the healthy group A and the colon cancer group B was statistically significant(P<0.01).The abundance of Fusobacterium nucleus in the colon cancer group was higher than the healthy group.Compared with the D Chinese medicine combined chemotherapy group,the B colon cancer group and C chemotherapy group were statistically significant(P<0.01).The abundance of Fusobacterium nucleus in the Chinese medicine combined chemotherapy group was significantly lower than the colon cancer group and chemotherapy group.It shows that oral Sijunzi Tang during chemotherapy can significantly reduce the abundance of Fusobacterium nucleatum in the intestine.There was no statistical significance between the A healthy group and the C chemotherapy group,the A healthy group and the D Chinese medicine combined chemotherapy group,and the B bowel cancer group and the C chemotherapy group(P>0.05).4.The TCM symptom score method was used to evaluate the improvement of the symptoms of spleen deficiency in the C chemotherapy group and the D Chinese medicine combined chemotherapy group.The effective rate of the Chinese medicine combined chemotherapy group was 86.67%.The total TCM symptom scores before and after treatment were 11.93 ± 2.59 and 7.17 ± 1.51,respectively,which were statistically significant(P<0.05).The effective rate of the chemotherapy group was 30.00%.The total scores of TCM symptoms before and after treatment were 11.00±1.93 and 10.00±2.12,respectively,which were not statistically significant(P>0.05).The comparison of the total scores of spleen deficiency symptoms between the two treatment groups was statistically significant(P<0.05).The KPS scores of the two treatment groups improved after treatment.The score of the C chemotherapy group was 66.33±5.56,and the score of the D Chinese medicine combined chemotherapy group was 72.33±6.79,the comparison between the two groups was statistically significant(P<0.05),indicating that Sijunzi Decoction combined with chemotherapy can significantly improve the symptoms of spleen deficiency and improve the quality of life of patients.The effective rate and stability rate of D Chinese medicine combined chemotherapy group was higher than the C chemotherapy group,but the difference in short-term objective curative effect was not statistically significant(P>0.05).5.In terms of safety evaluation,the number of cases of gastrointestinal reactions and peripheral neurotoxicity in the C chemotherapy group was significantly higher than the D Chinese medicine combined chemotherapy group.The comparison between the two groups was statistically significant(P<0.05),while myelosuppression occurred between the two treatment groups The situation was not statistically significant(P>0.05),indicating that Sijunzi Decoction can effectively reduce the adverse reactions of chemotherapy.Compared with before treatment,the liver and kidney functions of the two groups were not statistically significant(P>0.05),indicating that there was no liver and kidney damage during the treatment.Conclusion:1.Sijunzi Decoction combined with chemotherapy can increase the abundance of intestinal flora in patients with colorectal cancer,and normalize the diversity of intestinal flora.2.Sijunzi Decoction combined with chemotherapy can inhibit the growth of the pathogenic bacteria-Fusobacterium nucleatum.3.The treatment of colorectal cancer patients with Sijunzi Decoction during chemotherapy can effectively reduce the side effects of chemotherapy(digestive tract reactions,peripheral neurotoxicity),improve the symptoms of spleen deficiency,and improve the quality of life of the patients. |
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