Intestinal Flora Characteristics Of Patients With Colorectal Adenoma And Clinical Study On Preventing Recurrence Of Colorectal Adenoma After Operation With Qinghua-jianpi Decoction

Objective:1.Using 16S rRNA sequencing technology to detect colorectal adenomas patients and healthy people of intestinal flora,intestinal flora characteristics of treatment in patients with colorectal adenomas;2.Use of TCM qinghua-jianpi decoction intervention after colorectal adenomas adenoma resection patients,assessment of qinghua-jianpi decoction to preventing colorectal adenomas clinical effectiveness and safety of postoperative recurrence,and test analysis of qinghua-jianpi decoction on postoperative intestinal flora in patients with colorectal adenomas.Methods:Patients with colorectal adenomas structure of intestinal flora analysis:Twenty-one patients who accord with the diagnostic criteria for colorectal adenoma were selected as the experimental group,collected in the two groups patients basic information,Twelve healthy subjects with colorectal polyps excluded by colonoscopy were intaked as the control group.Basic information,BMI,preoperative hemoglobin albumin creatinine and other basic safety data of the two groups of patients were collected.Fecal samples of patients with colorectal adenoma and healthy controls were collected.Effect of qinghua-jianpi decoction on postoperative recurrence rate and intestinal flora in patients with colonic adenoma due to spleen deficiency and dampness.Forty cases of patients with spleen deficiency and dampness accumulation syndrome（TCM syndrome differentiation）were selected,whose stools were not returned to normal after 2 weeks of diet recovery.The patients were randomly divided into experimental group and control group with 20 cases in each group.Preoperative clinical symptom observation table was filled in and scored according to the quantitative grading standard of symptoms of spleen deficiency and dampness accumulation and the quantitative grading standard of colonoscopy pathological examination report.Basic information,BMI,preoperative hemoglobin albumin creatinine and other basic safety data of the two groups were collected.Feces samples of patients with colorectal adenoma postoperative before and after the intervention of qinghua-jianpi decoction were collected.Clinical trial:a randomized controlled study involving a total of 40 patients,patients 2 weeks after colorectal adenoma resection were randomly divided into treatment group（qinghua-jianpi decoction）and control group（placebo）,with 20 cases in each group.The clinical symptoms were observed at the time points of week 0,2 and 4.The clinical symptoms scores of the patients were recorded and analyzed at each time point.The colonoscopy was reviewed 6-12 months after the operation,and the postoperative colonoscopy pathology rating form was filled in.In the treatment group,fecal specimens were collected from 5 cases before intervention and 17 cases after intervention.Intestinal flora detection analysis:All collected fecal samples were sent to the high-throughput sequencing laboratory for intestinal flora detection.The characteristics of intestinal flora of patients with colorectal adenoma and the effect of qinghua-jianpi decoction on intestinal flora of patients with colorectal adenoma were analyzed by combining the data obtained from the test with clinical data.Result:1.Comparison of intestinal microflora structure between patients with colorectal adenoma and healthy controls.Baseline condition of subjects:There was no statistically significant difference in baseline data of Gender,age,BMI and Safety baseline data（hemoglobin、albumin、creatinine）between the 21 patients with colorectal adenoma included in this study and the 12 healthy controls（P>0.05）.Intestinal flora detection and analysis:a total of 33 qualified fecal samples were collected,including 21 samples from patients with colorectal adenoma and 12 samples from healthy controls.The two groups of samples were compared and the results showed that:OTU distribution,a total of 819 OTU samples were generated in the two groups,of which 636 were overlapped,OTU147 in the colorectal adenoma group and OTU36 in the healthy control group.Sparse analysis and Specaccum species-cum-curve analysis showed that the end curve gradually leveled off,indicating that the sequencing data volume was reasonable and the sample size was sufficient to meet the analysis standard.In terms of Alpha diversity index,chaol index observed_species（number of OTU）,PD_whole_tree index,Shannon index and other index differences were not statistically significant（P>0.05）.In terms of species distribution,at the phylum level of bacterial classification,Firmicutes dominated in both groups,followed by Bacteroidetes.The phylum with statistically significant difference between the two groups was Firmicutes、Bacteroidetes、Verrucomicrobia.At the class level of bacterial classification,in the group of colorectal adenomas,clostridia,Erysipelotrichia,Coriobacteriia and Verrucomicrobiae were dominate;In the normal healthy group,Bacteroidia and Negativicutes were dominantAt the order level of bacterial classification,Clostridiales、Erysipelotrichales and Coriobacteriales dominate in the colorectum adenoma group and Bacteroidales and Selenomonadales dominate in the healthy group were compared between the two groups.In terms of the family level of bacterial classification,the dominant family in the two groups were Lachnospiraceae,Erysipelotricaceae,Peptostreptococcaceae and Clostridiaceae₁,Coriobacteriaceae.The dominant species in the normal healthy groups were Bacteroidaceae and Porphyromonadaceae.From the bacterial classification of the genus level,the dominant bacteria of the colorectal adenoma group is the Blautia,Dorea,Holdemanella,Clostridium sensu stricto₁,Romboutsia,Collinsella.The normal healthy group was dominated by Bacteroides,Lachnoclostridium and ParabacteroidesThe core flora of the two groups was analyzed by LefSe.The core intestinal flora of the colorectum adenoma group was Firmicutes,Clostridiales,Clostridia,Lachnospiraceae,Blautia,Erysipelotrichia,Erysipelotrichaceae,Erysipelotrichales,Actin obacteria,Holdemanella;The core intestinal flora of the normal healthy group were Bacteroidia,Bacteroidales and Bacteroidetes,respectively.Bacteroides,Bacteroidaceae,Porphyromonadaceae,Parabacteroides,Negativicutes,Selenomonadales,Lachnoclostrid ium,etc.Combination can be thought of in a previous study Clostridium sensu stricto 1 could be one of the driving bacteria,Erysipelotrichaceae could be one of the passenger bacteria.2.A randomized controlled study of qinghua-jianpi decoction:There was no statistically significant difference between the two groups（P>0.05）in age,gender and disease severity（clinical symptom score before treatment,colonoscopy score,pathological score,and preoperative safety data）,which was comparable.The results showed that the treatment group was better than the control group in reducing the postoperative recurrence rate.The colonoscopy pathological score and clinical symptom score of the patients with spleen deficiency and dampness accumulation after colorectal adenoma surgery in the treatment group were all lower than those before treatment.Intra-group comparison:the scores of patients in the treatment group were compared at 2 and 4 weeks and at 0 weeks,and the differences were statistically significant（P<0.01）.The colonoscopy pathological score and clinical symptom score of patients with spleen deficiency and dampness accumulation after colorectal adenoma surgery were all lower than those before treatment.The difference of TCM syndrome points between the treatment group before intervention and after intervention was statistically significant（P<0.01）.The clinical symptom score of the control group was compared with that of the control group at 2 weeks and 4 weeks at 2 time points and 0 weeks.Although the score decreased,the difference was not statistically significant（P>0.05）.In the control group,the scores of TCM symptoms,loose stools and abdominal distension,decreased compared with that before treatment,and the differences were statistically significant（P<0.05）,while the differences of three TCM symptoms,abdominal pain,spiritlessness,poor appetite were not statistically significant（P>0.05）.Therefore,placebo may have a certain effect on loose stools and abdominal distension.Comparison between groups:there was no significant difference in the clinical symptom score between the treatment group and the control group before treatment,and the difference was statistically significant after 2 weeks and 4 weeks of treatment（P<0.01）.3.Comparison of intestinal flora before and after treatment with qinghua-jianpi decoction:The results showed that the fecal flora richness index and diversity index in the intestinal tract after colorectal adenoma surgery were significantly lower than that before colorectal adenoma surgery,with Chaol index and Shannon index both decreased,which was statistically significant.Compared with before the intervention,the difference of Chaol index was not significant after the treatment with qinghua-jianpi decoction,and the prognosis of Shannon index with qinghua-jianpi decoction was slightly higher than before the intervention,but there was no statistical difference after statistical analysis.In terms of species distribution,at the phylum level of bacterial classification,Firmicutes were decreased after colorectal adenoma resection,while the number of Firmicutes was not significantly increased by TCM,and there was no significant difference between the three groups.However,the number of Bacteroidetes increased slightly after the operation of colorectal adenoma,and the number of Bacteroidetes significantly doubled after the intervention of qinghua-jianpi decoction（p=0.01）,showing a statistical difference.Proteobacteria increased significantly after colorectal adenoma resection,while the preoperative level was restored after the intervention of TCM.The preliminary consideration was that in the postoperative recovery process,the mucosal flora transferred and colonized into the intestinal cavity at a faster rate,resulting in a relatively large fluctuation,and the possibility of more proteobacterium colonization in the intestinal mucosa could not be excluded.Actinobacteria was significantly reduced after colorectal adenoma surgery,and the number of Actinobacteria was not significantly increased after TCM intervention,indicating that intestinal preparation and intestinal polypectomy had a great impact on actinomycetes,while the effect of TCM intervention was not obvious.Fusobacteria increased significantly after colorectal adenoma surgery（p>0.05）,and there was no statistical difference.The possible reason was that there were significant differences among individuals in the group,which affected the statistical structure.There was no significant increase after TCM intervention,indicating that TCM intervention had no significant effect on FusobacteriaAt the genus level of bacterial classification,the group of colorectal adenomas and the group of colorectal adenomas after surgery+TCM.Statistically significant,and the bacteria that have reduced their populations are Blautia,Klebsiella,Streptococcus,Lactobacillus,Eubacterium hallii_group,Dorea,Ruminococcus_torques_group,Holdemanella,Clostridium_sensu_stricto₁ and Romboutsia,Fusicatenibacter,Collinsella,erysipelotrichaceae_ucg-003;It is statistically significant that the bacterium number has increased in the genus Prevotella₉,Phascolarctobacterium and Veillonella.Postoperative colorectal adenomas compared before and after the intervention,Chinese traditional medicine have statistical significance and to reduce the number of bacteria of Escherichia-Shigella and to increases the number of bacteria of Lachnoclostridium,it can be seen that TCM can be targeted to adjust the number of intestinal flora,reach the purpose of regulating intestinal flora and Escherichia-Shigella bacteria is one of the starting bacteria of colorectal cancerBy constructing evolutionary trees,The main dominant bacteria in the group of colorectal adenomas were c Bacilli,c_Clostridiales,c_Erysipelotrichia and p_Actinobacteria.The dominant bacteria in the operation group of colorectal adenoma were mainly p_Proteobacteria;P_Bacteroidetes was the dominant bacteria in the group of colorectal adenoma surgery and intervention with TCM.LefSe analysis.In terms of the genus level of bacterial classification,The dominant bacteria in the colorectal adenoma group were Blautia,Lactobacillus and Streptococcus,Holdemanella,Eubacterium hallii_group,Ruminococcus_torques_group,Clostridium sensu_stricto₁,etc;The main bacteria in the intestinal tract after colorectal adenoma surgery were Escherichia_coli,Veillonella,Klebsiella etc;The dominant bacteria in the TCM intervention group after colorectal adenoma surgery were Bacteroides,Prevotella₉,Phascolarctobacterium,Lachnoclostridium etc.Conclusion1.There was no significant difference in the Alpha diversity index between the patients with colorectal adenoma and the healthy control group,and the species distribution of the intestinal flora was basically the same as that of the healthy control group.The abundance of some strains was different from that of healthy controls:At the phylum level,the relative abundance of the microflora of firmicutes,Actinobacteria and Verrucomicrobia was higher than that of the healthy control group,while that of Bacteroidetes was lower than that of the healthy control group.At class level,in the group of colorectal adenomas,the dominant group is Clostridia,Erysipelotrichia,Coriobacteriia and Verrucomicrobiae.In the normal healthy group,Bacteroidia and Negativicutes were dominant.At the order level,Clostridiales,Erysipelotrichales and Coriobacteriales were dominant in the colorectoma group,and Bacteroidales and Selenomonadales were dominant in the healthy group.At the family level,the dominant bacteria families in the colorectoma group were Lachnospiraceae,Erysipelotrichaceae,Peptostreptococcaceae,Clostridiaceae₁,and Coriobacteriaceae;the dominant groups in the normal healthy groups were Bacteroidaceae and Porphyromonadaceae.From the genus level,the dominant species of colorectal adenomas are Blautia,Dorea,Holdemanella,Clostridium_sensu_stricto₁,Romboutsia and Collinsella.The normal healthy group was dominated by bacteroides,Lachnoclostridium and Parabacteroides.Combination can be thought of in a previous study Clostridium_sensu_stricto₁ could be one of the driving bacteria,Erysipelotrichaceae could be one of the passenger bacteria.2.Qinghua-jianpi decoction can reduce the recurrence rate of postoperative patients with colorectal adenoma,improve the clinical symptoms of patients with spleen deficiency and dampness accumulation syndrome,and have a certain impact on the intestinal flora of patients with colorectal adenoma.Escherichia-shigella decreased and Lachnoclostridium increased after TCM intervention for colorectal adenoma Postoperative.It is possible to prevent recurrence of colorectal adenoma by regulating the abundance of Escherichia-Shigella and Lachnoclostridium bacteria.