Intestinal Microbial Diversity And Clinical Relevance Analysis On Patients With Wilson Disease

Objective1.1 V4 hypervariable region of 16 S rRNA of intestinal microorganism from Wilson disease patients and healthy people will be sequenced and analyzed, based on Illumina PE250 sequencing platform; Data statistics and analysis will be done by bioinformatics analysis method for measured gene sequence, to observe intestinal microorganism diversity and structural characteristics of Wilson disease.1.2 To compare the correlation between intestinal bacterial abundance and clinical parameters in patients with Wilson disease, and to explore the mechanism of intestinal micro ecological imbalance on the occurrence and development of the disease.Method2.1 Select 22 cases of patients with Wilson disease as the case group, select 22 healthy people as the control group, and collect the feces samples of the objects.;2.2 Use column fecal DNAout kit to extract DNA of fecal bacteria population from the object group, then do the PCR amplification, Illumina PE250 library construction and 16 S rRNA V4 hypervariable region sequencing;2.3 Detect copper biochemistry in patients with Wilson disease by immunoassay,and detect trace elements(Cu, Fe, Zn) in serum and feces of patients with Wilson disease by using flame atomic absorption spectrometry(FAAS).Result3.1 Compared with the control group, the Shannon index of intestinal microorganism community decreased significantly in Wilson patients(2.61 vs2.53,P<0.05);3.2 In the phylum level, the analysis shows that the intestinal Proteobacteria(4.31%vs 3.09%,P<0.001),Tenericutes(0.23% vs 0.19%,P<0.001)and Fusobacteria(1.88%vs 0.04%,P<0.001)and Firmicutes(26.18% vs 19.83%,P<0.001)bacterial abundances of Wilson case group were significantly higher than those of the normal control group,while Bacteroidetes abundance decreased significantly(67.19% vs 76.75%,P<0.001);3.3 In the genus level, Bacteroides(48.5% vs 4.6%, P<0.001),Faecalibacterium(2.9% vs 2.1%,P<0.001),Megamonas(0.8% vs 0.2%,P<0.001),Roseburia(0.7%vs 1.3%, P<0.001) abundances of Wilson case group were significantly higher than those of the normal control group, while Prevotella abundance decreased significantly(1.6% vs 2.4%,P<0.001);3.4 The contents of Cu, Fe and Zn trace elements in the feces of Wilson cases were significantly lower than those in the control group(Cu,T=-3.816,P<0.001;Fe:T=-5.289,P<0.001;Zn:T=-2.704,P<0.001);3.5 In the phylum level, Firmicutes abundance in patients with Wilson disease was negatively correlated with Cu concentration in the feces(R=-0.044,P=0.041<0.05), and Bacteroidetes abundance was positively correlated with Cu concentration(R=0.552,P=0.008<0.01);3.6 In the genus level, intestinal Bacteroidetes abundance in patients with Wilson disease was negatively correlated with the rating(R=-0.571,P=0.006<0.01).Conclusion4.1 The intestinal microorganism diversity in patients with Wilson disease decreased significantly than that in the normal group;4.2 In the phylum and genus levels, the intestinal microorganism structure of patients with Wilson disease is significantly different from the normal people;4.3 The intestinal Firmicutes and Bacteroidetes abundances in patients with Wilson disease are related to the Cu content in feces;4.4 The intestinal Bacteroidetes abundance in patients with Wilson disease is probably related to the severity of clinical symptom.