Study On Intestinal Microecology Of Infants With Cholestatic Liver Disease

Objective: By using 16 s rDNA Amplicon sequencing technique and bioinformatics analysis,the changes of intestinal microbial community in children with infantile cholestatic liver disease were studied.Methods: From December 2016 to January 2018,43 children with infantile cholestatic liver disease who were hospitalized in the Department of Digestive Diseases of Children's Hospital of Hebei Province and the Eastern Hospital of the second Hospital of Hebei Medical University were selected as the case group.In the same period,37 infants in the pediatric outpatient clinic of the third Hospital of Hebei Medical University were selected as the control group.Both groups chose Shijiazhuang City or its surrounding counties,their eating habits and lifestyle.The selected infants obtained informed consent from their parents,signed informed consent letters,and were examined and approved by the Medical Ethics Committee of the third Hospital of Hebei Medical University.Stool samples were collected from both groups.Data were obtained by 16 s rDNA sequencing and bioinformatics analysis was carried out.Result:1.Data information: totally 7056085 Raw Tags were obtained from 43 cases of fecal specimens and 37 cases of control group(control group)by sequencing.6738507 Effective Tags were obtained after data quality control.The total base number was 1702232693 BP with an average of252.83 bp.After clustering,the total number of OTUs was 22.320,and the average OTUs number per sample was 279.2.Species notes: species composition and abundance A total of 30 phylum were detected at the gate level.More than 99.69% of the detected phylum were the following four phylum: Firmicutesa,Actinobacteriae,Proteobacteria,Bacteroidetes.The abundance of phylum thuringiensis in the case group and control group was 0.4179 and 0.4285 respectively,the abundance of actinomycetes in the case group and control group was 0.2214 and 0.2846,and the abundance of Proteus in the case group and control group was 0.1.2531 and 0.1372,the abundance of Bacteroides in case group and control group were 0.1045 and 0.1476,respectively.The fecal abundance of the two groups was the highest,and the fecal abundance of the patients in the case group was lower than that in the control group.The fecal abundance of the faeca,actinomycetes and Bacteroides in the case group was lower than that in the control group.A total of 349 genera were detected at the generic level.The genus top10 is Bifidobacterium ? Escherichia-Shigella ? Enterococcus ? Bacteroides ?Streptococcus ? Staphylococcus ? Veillonella ? Lactobacillus ? Blautia and Faecalibacterium tenella.Bifidobacterium abundance was higher in the faeces of both groups.Compared with the control group,the fecal Bifidobacterium,Bacteroides,Blauterium,Veronella and Clostridium tenella in the case group were low in abundance,while Escherichia coli and Enterococcus were found in the feces.Streptococcus,Staphylococcus and Lactobacillus belong to 5species of high abundance.3.Rarefaction Curve: in the 80 samples tested,when the amount of sequencing increases,the curve tends to be flat.It shows that the samples obtained in this study are reasonable and the depth of sequencing is enough,which can be further analyzed.4.Species accumulation boxplot: in the 80 samples measured,with the increase of sample size,a large number of species were found,and the curve showed a sharp upward trend;as the sample size continued to increase,the curve gradually flattened.It indicates that the species in the sample will not increase significantly with the increase of sample size,which indicates that the sample size in this study is sufficient and can meet the needs of sampling.5.NMDS analysis: the samples of case group and control group formed clusters on the two-dimensional plane,which indicated that the speciescomposition similarity of each group was high,and the similarity of species composition between the two groups was different.That is,the two groups of species composition is comparable.6.LEfSe analysis: there was significant difference between the two groups(P<0.05,LDA>4).Compared with the control group,the feces of the two groups had higher abundance of Proteobacteria and Actinobacteria.Compared with the control group,the feces of the two groups had higher abundance of Proteobacteria and lower abundance of Actinobacteria,Bifidobacterium,Enterococcus,Streptococcus,Staphylococcus,Blauterium and Clostridium tenella were significantly different between the two groups at the generic level(P < 0.05 LDA > 4).Compared with the control group,the faecal bifidobacterium,Blauteria brucellae and C.tenella in the case group were compared with those in the control group.The abundance of Clostridium tenella was lower than that of Enterococcus,Streptococcus and Staphylococcus.Conclusion:1.Differences in intestinal microbial composition between children with cholestatic liver disease and healthy children.2.Based on the results of 16 s rDNA sequencing and bioinformatics analysis,it was found that more than 99% of the intestinal microflora in children and healthy infants with cholestatic liver disease were mainly phylum pachymatidis,actinomycetes,Proteus,Bacteroides.Bifidobacterium,Escherichia coli,Enterococcus,Bacteroides,Streptococcus,Staphylococcus,Veronella,Lactobacillus,Blauterium and Clostridium tenella.3.Diversity and abundance of intestinal microorganism in infants with cholestatic liver disease: at the portal level,the abundance of actinomycetes in intestinal microbes of infected infants decreased,while that of Proteus increased.At the generic level,the abundance of bifidobacterium,brucellosis and Clostridium tenella in the intestinal microbes of sick infants decreased,and the abundance of opportunistic pathogens such as Enterococcus,streptococcus and Staphylococcus increased,and the intestinal microecologywas out of balance.The overgrowth of opportunistic pathogens may be involved in the occurrence and development of the disease.