Experimental And Clinical Study On The Relationship Between Gastrointestinal Fever Syndrome And Intestinal Microecological Imbalance In Children

Gastrointestinal Heat Retention Syndrome is the syndrome of intangible or tangible heat in the stomach and intestine which is more frequently occurred on the infants than the adults. The objective of this study is to explore the relationship between this syndrome and the intestinal microbiome disorder on the infants by testing the mice’s and children’s intestinal microbiome with the method of High-throughput sequencing which makes the research of pathogenesis more comprehensive and provides a new perspective for the modern interpretation of the syndrome.Literature review, experimental study and clinical study are involved.Part One Literature ReviewMethod:In this part, the pathogeny, pathogenesis and the reason of its frequent occurrance in modern society of the Gastrointestinal Heat Retention Syndrome are summarized through reviewing and consulting the ancient and modern literatures as well as the application prospect of systems biology in this field; on the other side, the constitutes and functions of the intestinal microbiome are elaborated as well as the factors that can make influence on individual characteristics of its balance and imbalance, and the manifestation of the intestinal microecology disorder and the relevance between the disorder and the diseases are also expatiated along with its future perspective in the Traditional Chinese Medicine.Results and Conclusion:The literature research indicates that eating more high-fat, emotional frustration and exogenous evil cause the Gastrointestinal Heat Retention Syndrome, the modern pathogenesis associates it with immunologic derangement, neuroendocrine disorders and alogotrophy; in nowadays, some factors such as the eutrophia dietary, the food safety issues, the mental pressure and the lack of movement make the Gastrointestinal Heat Retention Syndrome more common. Intestinal microbiome can regulate the nutrient metabolism, immunostimulation and biological antagonistic of the host as well as show a individual characteristic in infants with different delivery or feeding patterns and adults with different age, region and sports habits. Meanwhile, gut microbiome disorder can be observed when exogenous（climate, diet, environmental pollution, pathogenic microorganism and food additives）, endogenous（nutritional metabolic defects, dysfunction of organs and cancer） and iatrogenic（drugs, radiotherapy, chemotherapy and surgery） factors change in the proportion and the parasitic site of microorganisms in the intestinal which result in the diseases of digestive system, metabolism and anaphylaxis. The study of intestinal microbiome establishes a new perspective for the modernization of Traditional Chinese Medicine.Part Two Experimental StudyObjective:This study is to explore the relevance between the Gastrointestinal Heat Retention Syndrome and the intestinal microbiome disorder by detecting the colorectal content samples from the syndrome model and normal control group of mice and make the experimental study on pathogenesis of the syndrome more consummate.Method:Make the syndrome model mice by feeding special fodder which is deemed to be a high protein and calorie diet and collect the colorectal content samples in a germ-free environment. Detact the diversity and absolute/relative abundance of the intestinal microbiome in the samples with the method of High-Throughputing Sequencing, describe the structure characteristics of the intestinal microflora on mice, excavate the differences between groups, and screen the special gut microecological index relating with Gastrointestinal Heat Retention Syndrome on mice.Results:1 Mice’s general signs of observationThe normal control group（NC） are in good mental state and physical agility which have white soft glossy furs and brownish black formed stools.The model group （M） are in poor mental state and physical agility which have yellowish somber furs and pale brown soft sticky stools.2 Mice’s weights observationThe model group are lighter on weights than the normal controls in the day to draw materials.3 The structure characteristics of intestinal microbiome on miceThe intestinal microbiome on mice presents diversification with 9 phylum、16 classes、 30 orders、46 families.57 genuses and 26 species are involved. The most OTUs are annotated on the family level and the dominant bacteria are Lachnospiraceae, Bacteroidaceae, S24-7 and Ruminococcaceae.4 The comparison on intestinal microbiome between the model mice and normal control miceâ‘  The PCA and PCoA result in the Beta diversities analyze indicate that the structure of the intestinal microbiome on the model group are different from that on the normal control group;â‘¡ The diversities of the intestinal microbiome are different between the NC female mice and M female mice on the Rarefaction and Chol 1 Curve while the diversities on the genus level and species are different between the model and the normal control group;â‘¢ The reduction of the relative abundance of Lachnospiraceae is obvious on the model mice while the enrichness of Bacteroidaceae is obvious on female mice;5 The screening of the special gut microecological index with Gastrointestinal Heat Retention Syndrome on miceOTU₁ （the annotation is p_Firmicutes; c_Clostridia; o_Clostridiales; f_Lachnospiraceae） is the characteristic index screened by Venn、Heatmap and the OTUs relative abundance which decreases obviously on the model group.Conclusion:1 The model is successful according to the changes of mice’s symptoms and signs.2 The intestinal microbiome on mice presents diversification and the dominant bacterias on the family level are Lachnospiraceae, Bacteroidaceae, S24-7 and Ruminococcaceae.3 The intestinal microbiome of the model group is disorder which indicates that Gastrointestinal Heat Retention Syndrome is interrelated with the intestinal microbiome disorder.4 The characteristic intestinal microbiome index of the Gastrointestinal Heat Retention Syndrome on mice is OTU₁ which decreases obviously on the syndrome group.Part Three Clinical StudyObjective:This study is to explore the relevance between the Gastrointestinal Heat Retention Syndrome and the intestinal microbiome disorder by detecting the feces samples from the syndrome and healthy children and make the clinical study on pathogenesis of the syndrome more consummate.Method:Collect the feces samples from children according the （（Diagnostic Scale of Gastrointestinal Heat Retention Syndrome）） and detact the diversity and absolute/relative abundance of the intestinal microbiome in the samples with the method of High-Throughputing Sequencing, describe the structure characteristics of the intestinal microflora on children, excavate the differences between groups, and screen the special gut microecological index relating with Gastrointestinal Heat Retention Syndrome on children.Results:50 children are involved in this study with 26 in Gastrointestinal Heat Retention Syndrome Group（with 14 male and 12 female） and 24 in Healthy Group（with 12 male and 12 female）.1 General informationThere is no significant difference between Gastrointestinal Heat Retention Syndrome children and healthy children on average age, age stratification. BMI（Body Mass Index）, the way of childbirth, the pregnancy time period, the history of vaccination, the situation of probiotic dairy drinking or diets habits.2 The structure characteristics of the intestinal microbiome on childrenThe intestinal microbiome on children presents diversification and 20 phylum,30 classes, 65 orders,105 families,156 genus and 66 species are included. The most OTUs are annotated on the genus level and the dominant bacteria are Bacteroides and Faecalibacterium.3 The comparison on intestinal microbiome between Gastrointestinal Heat Retention Syndrome children and healthy childrenâ‘  There is no significant difference between the two groups on the intestinal microbiome structure with the PCA and PCoA;â‘¡ There is no significant difference between the two groups on the intestinal microbiome diversities on the Rarefaction and Chol 1 Curve on the genus and species level;â‘¢ The relative abundance of Bacteroides and Faecalibacterium are identical between the two groups;4 The screening of the special gut microecological index with Gastrointestinal Heat Retention Syndrome on childrenThere is no characteristic index relating with Gastrointestinal Heat Retention Syndrome on children according to Venn, Heatmap and the abundance of OTUs.Conclusion:1 The intestinal microbiome on children presents diversification and the dominant bacteria on the level of genus are Bacteroides and Faecalibacterium.2 The reason that there is no difference between the Gastrointestinal Heat Retention Syndrome children and healthy ones on the intestinal microbiome structure may be that: â‘  the intestinal microbiome is still in the self regulation limit when the syndrome is mild; â‘¡the location of the syndrome is stomach and large intestine while the intestinal microbiome disorder may be caused by imbanlance of two or more systems;â‘¢the syndrome maybe relates to the mocusa-associated bacteria while the bacteria in the feces samples are mainly living in the lumen;â‘£the influence from the probiotic dairy on the intestinal microbiome is much greater than that from the syndrome.