| Part 1 The diversity and similarity of microbiome spectrum in saliva、induce sputum and fecal from bronchial asthmaBackground:Bronchial asthma is an chronic airway inflammatory disease which is involved in a variety of inflammatory cells.The process of disease is closely related to heredity,environment,immune status of organism while microbiome play an important role in it.According to the development of 16 sr RNA detection,pelple found that the diversity of bacterial flora in oral cavity、respiratory tract and digestive tract were different among healthy people but they also have different degrees of similarity.It demonstrated that microbiome from different site of human body interact with each other.The difference and similarity of microbiome structure in oral cavity、respiratory tract and digestive tract from asthmatic and its relationship with asthma are not clear.Objective:Comparing with the diversity and similarity of microibome spectrum in saliva 、induce sputum and fecal from asthmatic.Methods:We recruited 55 asthmatic,collecting their saliva 、 induce sputum and fecal for16 sr RNA analysis at the same time.There are 32 asthmatic’s eligible sample were accepted to detect finally.V4 regions of bacterial 16 S r RNA genes were amplified and sequenced by Miseq 250.Sequences were assigned to bacterial taxa by comparing them with 16 s r RNA sequences in the Ribosomal Database Project.Result:1.Compared to induce sputum,fecal had higher raw read count and effective read count(p<0.05).Induce sputum had highest effective read rate,the next was fecal,saliva had lowest effective read rate(p<0.05).The total OTU of fecal higher than saliva and induce sputum significantly.2.In aspect of Alpha diversity,Shannon index of saliva higher than fecal(p<0.05).Simspon index of Induce sputum higher than fecal(p<0.05).3.According to Beta diversity,the diversity of bacterial flora between saliva and fecal was highest,the next was between induce sputum and fecal,the diversity of bacterial flora between saliva and induce sputum was lowest(R2=0.5081,p<0.001).4.The index of similarity between saliva and fecal was highest,the next was induce sputum and fecal,the index of similarity between saliva and induce sputum was lowest.The index of similarity about saliva-fecal was higher than induce sputum-fecal and saliva-induce sputum significantly(p<0.05).Conclusion:The diversity of bacteria in saliva 、 induce sputum and fecal from asthmatic are different while all of them interact with each other in different degree.Bacterial community structure and relative abundance of saliva and fecal have the highest similarity.Part 2 The analysis of microbiome spectrum in induce sputum and fecal and its correlation with clinical featrues of bronchial asthmaBackground:The human respiratory tract was formerly thought to be in a sterile environment unlit the development of micorbiome gene detection.Lots of research indicated that there was a difference in the diversity of respiratory tract bacterial flora between healthy subjects and asthmatics,which was correlated with clinical features of bronchial asthma.The hygiene hypothesis suggesting that the change of composition and diversity of microbiome in intestinal tract in early life is a risk factor for the development of adultasthma.So there is closely relationship between microbiome in airway and gut and asthma.However,most of these researchs were from foreign countries.There has been very little information available from cohorts of asthma in china.Objective:To comapre the difference of bacterial spectrum in induce sputum and fecal between asthmatic and healthy people and its correlation with clinical features of asthma.Methods:Sputum test: recruiting 41 asthmatic and 22 healthy people for 16 sr RNA analysis.Fecal test: recruiting 38 asthmatic and 24 healthy for 16 sr RNA analysis.All participants underwent clinical assessment,spirometry、routine blood test 、allergen detection and sputum induction.A total of 37 qualified induce samples(24 asthmatic and 13 healthy separately)and 44 qualified fecal sapmles(29 asthmatic and 15 healthy separately)were sequenced for 16 S r RNA analysis.The V4 regions of bacterial 16 S r RNA genes were amplified and sequenced by Miseq 250.Sequences were assigned to bacterial taxa by comparing them with 16 s r RNA sequences in the Ribosomal Database Project.Taking correlation analysis between microbiome of sputum and fecal and clinical features of asthma.Results:1.Sputum test:(1)compared to healthy people,asthmatic were older[50.0(40.6,52.3)vs 27.5(26.3,41.9),p=0.009]and had higher percentage of sputum eosinophils[9.3(11.9,31.7)vs 0.9(0.3,2.2),p<0.001]、blood eosinophils[4.6(3.7,7.0)vs 1(0.5,3),p<0.001]、co unt of blood eosinophils[0.3(0.2,0.5)vs 0.1(0,0.2),p<0.001] and total Ig E[172.0(124.8,486.9)vs 25.2(8.7,85.7),p<0.001],worse lung function(FEV1%pred for example,77.4±3.9 vs 94.1±2.2,p<0.001).(2)There was no significant difference in Alpha diversity between asthmatic and healthy people.(3)At phylum、Class and Order level,Fusobacterium were present significantly lower abundance in induce sputum from asthmatic(p<0.05)(4)There are no significantly correlation between the spectrum in induce sput-um and features of asthma.2.Fecal test:(1)Compared to healthy people,asthmatic were older(47.7±2.5vs36.1±3.9,p=0.013)and had higher percentage of sputum eosinophils[12.4(13.2,29.0)vs 0.8(0.3,2.0),p<0.001]、blood eosinophils[4.4(3.2,6.5)vs 1.6(0.95,2.09),p<0.001]、count of blood eosinophils[0.3(0.2,0.4)vs 0.09(0.06,0.12),p<0.001] and total Ig E[158(89.1,549.9)vs 17.2(-44.6,151.3),p<0.001],worse percentage of sputum macrophag-e[16.6(16.5,34.3)vs 47.5(28.4,60.2),p=0.05] and lung function(FEV1%pred for example,72±3.8vs95±2.2,p<0.001)(2)There was no significant difference in Alpha diversity between asthmatic and healthy people.(3)At phylum level,the abundance of Tenericutes was higher in fecal from healthy people(p<0.05).At f-amily level,Acidaminococcus and Lachnospira wa-s significantly common in asthmatic and healthy people respectively(p<0.05).(4)The abundance of Acidaminococcus had positively correlated with FEV1(R=0.391,p=0.036)and MEF75%(R=0.658,p=0.001).The abundance of Lach nospira had negative correlated with both the percentage of sputum eosinophils(R=-0.420,p=0.019)、count of blood eosinophils(R=-0.465,p=0.013)and dose of ICS(R=-0.417,p=0.024).Conclusion:Compared with the healthy control group,bronchial asthma has different micr-obiome spectrum in respiratory tract and intestine tract.The bacterial spectrum in fecal are related to clinical features of asthma,but there are not significant correl-ation between the bacterial spectrum in induce sputum and clinical features of ast-hma. |
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