| Objective:Gastrointestinal heat is a sign of visible heat knot or invisible heat pathogen stagnant in the stomach and intestines.Because the children's viscera is tender and the spleen and stomach are weak,they tend to suffer from food accumulation,and then accumulate stagnation and become hot,forming a syndrome of heat accumulation in the stomach and intestines.Gastrointestinal heat is closely related to the incidence and prognosis of pneumonia in children.It has been found that gastrointestinal fever can cause intestinal flora disturbance,which is related to the local immune function of intestinal mucosa and the level of immunoglobulin and inflammatory factors.Based on previous studies,we studied the changes of intestinal microecology as an important condition between gastrointestinal heat and pneumonia.We used molecular biology and high throughput sequencing to detect the changes in receptor pathway proteins and inflammatory factors in intestinal microflora associated with intestinal microflora in young rats.We used animal characterization,gastrointestinal motility,inflammatory factor levels,protein expression and bacterial community structure as the main indicators.We explored the relationship between gastrointestinal heat,pneumonia and intestinal microecological disturbance in children.We expect to explain the biological mechanism of children's gastrointestinal fever and pneumonia through intestinal flora,and improve the related pathogenesis of gastrointestinal fever and pneumonia in children,and discuss the scientific connotation.This study provides experimental evidence for the study of gastrointestinal fever and pneumonia in children,and has practical significance for improving the clinical efficacy of infantile gastrointestinal fever complicated with pneumonia.Method:The gastrointestinal heat accumulation model of young rats was prepared by intragastric administration of high calorie feed combined with milk.We reproved the Chinese patent medicine called "Xiao Er Hua Shi Wan" We observed animal signs,viscera coefficient and intestinal pathology,and analyzed the correlation between the data.We used high calorie feed combined with milk and gavage combined with LPS atomization inhalation to prepare gastrointestinal tract fever complicated with pneumonia in young rats.We observe animal signs and histopathology,calculate viscera coefficient,use Aimplex technique to detect serum inflammatory factors,detect intestinal tissue pattern recognition receptor protein by easy Western technique,detect intestinal microecological disturbance by high throughput technique,and further analyze the correlation between the data.Result:1.general condition of gastroenteric feverIn the gastrointestinal heat group,the rats were depressed,reduced in activity,discolored in hair,yellow in urine,dark in stool,and gradually shiny and oily.After feeding the children's food pill,the general condition of the animals improved.Compared with the normal group,the weight loss and abdominal circumference of the rats in the GI group had no difference(P>0.05),and the diet decreased,the difference was statistically significant(P<0.05);the number of fecal particles decreased,the weight of the stool decreased,the axillary temperature decreased,and the anal temperature decreased(P<0.1).After the treatment,the weight loss,the decrease of diet and the change of abdominal circumference were not different,the difference was not statistically significant(P>0.05),the number of fecal particles decreased,the weight of the stool decreased,the axillary temperature rose,and the anal temperature increased,and the difference was statistically significant(P<0.01).2.viscera of gastroenteric feverCompared with the normal group,the index of liver,spleen and kidney decreased and the index of lung increased,but the difference was not statistically significant(P>0.05).In the gastrointestinal heat accumulation group,the intestinal mucosa became thinner,the number of goblet cells decreased,and the submucous lamina propria was hyperemia,edema and fibrous hyperplasia.After treatment with infantile food pill,the goblet cells arranged orderly and the glands gradually recovered.3.gastrointestinal motility animal serum gastric motility hormoneThe gastrin decreased in the GI group,and the gastrin level of the group was increased after the combination of the treatment of the children,and the difference was statistically significant(P<0.05).There was no significant difference in serum motilin level(P>0.05).4.animal general situation of gastrointestinal accumulation and pneumonia Compared with the normal group,the rats in the group of gastroenteric heat combined with pneumonia had the phenomenon of reduced activity and piled up,the hair color was disheveled,the abdomen was bulging,the urine was deep yellow,the color of the stool was brown,the oil was light,and even the greasy loose stool appeared.Compared with the pneumonia group,the rats with gastrointestinal heat and pneumonia had gastrointestinal symptoms of deep heat,dark brown stool,brown and oily appearance.Compared with the normal group,the body weight and Anal temperature decreased in the rats with gastrointestinal fever and pneumonia,but the difference was not statistically significant(P>0.05).The difference of abdominal circumference and axillary temperature was statistically significant(P<0.05).Compared with the group of pneumonia group,the body weight loss,abdominal circumference,Anal temperature and axillary temperature decreased in the group of gastrointestinal accumulated fever and pneumonia group,and the difference was statistically significant(P<0.05).5.gastrointestinal fever complicated with pneumonia,animal organs and serum gastric motility hormonesCompared with the normal group,the lung coefficient increased and the liver coefficient decreased in the group of GI combined with pneumonia,and the difference was statistically significant(P<0.05).The coefficient of spleen and kidney increased,but the difference was not statistically significant(P>0.05).Compared with pneumonia group,the lung weight coefficient of the rats with gastrointestinal heat accumulation and pneumonia group decreased,the difference was statistically significant(P<0.05).In the rats with gastrointestinal fever and pneumonia,the colonic tissue was clear and intact,and the surface of the colonic mucosa was smooth without villi.The epithelial structure of the colonic mucosa is incomplete,and the monolayer columnar epithelial cells are arranged closely and neatly.There are many intestinal glands in the lamina propria,and the intestinal glands are straight and long and closely arranged.There is a large increase of goblet cells in the intestinal mucosa.There were no pathological changes of hyperemia,edema,inflammatory cell infiltration and so on.Pulmonary alveolar structure was not clear,pulmonary septum widened significantly,pulmonary alveolar fusion even disappeared,vascular dilatation and congestion,accompanied by a large number of inflammatory cells infiltration.Compared with the normal group and the pneumonia group,the serum gastrin decreased in the group of gastrointestinal accumulated fever and pneumonia group.The difference was statistically significant(P<0.05),and the difference in motilin change was not statistically significant(P>0.05).6.gastroenteric fever with pneumonia,gastrointestinal heat and pneumonia in animals,TLRs pathway.Compared with the normal group,the serum KC decreased in the group of GI and pneumonia group,and the difference was statistically significant(P<0.05);IL-12,IL-6 and TNF-alpha were increased,but the difference was not statistically significant(P>0.05).Compared with the pneumonia group,the serum KC increased in the GI group and the pneumonia group,and the difference was statistically significant(P<0.05);IL-12 decreased,IL-6,TNF-alpha increased,but the difference was not statistically significant(P>0.05).Compared with the normal group,the expression of MyD88,MD2,NF--kappa B,IkB-alpha and TLR9 expression in the intestinal tissue of the group of GI combined with pneumonia was increased,but the difference was not statistically significant(P>0.05).Compared with the pneumonia group,the expression of TLR9 in the intestinal tissue of the rats with gastrointestinal fever combined with pneumonia was significantly increased(P<0.05).The expression of MyD88,MD2,NF-,B and IkB alpha was decreased,but the difference was not statistically significant(P>0.05).7.intestinal microflora combined with intestinal microflora of pneumonia in animalsThere were significant differences in the overall classification structure of gastrointestinal tract fever with pneumonia and normal young rats,gastrointestinal fever rats and pneumonia rats.The group of gastrointestinal accumulated fever and pneumonia group was 60.Compared with the normal control group,there were 458 groups of gastrointestinal accumulated fever and pneumonia group.Compared with the gastrointestinal accumulation group,there were 641 groups of gastrointestinal accumulated fever and pneumonia group.Compared with the pneumonia group,there were 521 group of gastrointestinal accumulated fever and pneumonia group.The species annotation information of pneumonia rats includes 6 species of fungi,12 species,18 species,35 species,38 fungi and 19 strains.Compared with the normal group,the lactobacillus in the group of gastroenteric heat combined with pneumonia was significantly reduced.Compared with the Staphylococcus and the dynamic gastroenteric heat group,Staphylococcus,Staphylococcus,Staphylococcus,Staphylococcus,Bacillus,Bacillus,Balloon and Twinning were significantly increased in the group of Staphylococcus and pneumonitis.Compared with the pneumonia group,the gastrointestinal tract was significantly increased.Corynebacterium,Corynebacaceae and Actinomycetes were significantly reduced in heat accumulation combined pneumonia group.Based on the analysis of inflammatory structure,the relationship between IL-12 and IL8 was significantly greater than that of IL-6 and TNF-alpha.Among them,TNF--IL-6 and IL-6-IL8 are positively correlated,and TNF-negative-IL8,TNF-alpha-IL-12,IL-6-IL-12 and IL8-IL-12 are negatively correlated.Network inference based on the relationship between microbial members suggests that there may be a "competitive" relationship between Lactobacillus-Dorea and Lactobacilli-Rumen.The metabolic function of bacteria groups is predicted,involving the degradation and metabolism of exogenous substances,metabolism of nucleotides,terpenoids and cluster metabolism,other amino acid metabolism,metabolism of cofactors and vitamins,metabolism of lipid compounds,polysaccharide biosynthesis and metabolism,enzyme family,energy metabolism,carbohydrate Metabolism and other secondary metabolites are biosynthesis and amino acid metabolism.Conclusion:The animal model of gastro intestinal fever prepared by self-made high calorie feed combined with milk gavage method conforms to the clinical characteristics.The model animals have nutritional and gastrointestinal motility changes,but there is no obvious inflammatory damage and no malnutrition in the intestinal tissue.The gastrointestinal heat caused by improper diet can affect the body's nutrition absorption,cause intestinal probiotics disorder,and affect the immune function of the lung.Pneumonia animal will appear the probiotics reduces,the pathogenic bacteria increases the intestinal flora disturbance.The intestinal microflora of gastrointestinal fever patients complicated with pneumonia includes the competition of probiotics and the more pathogenic ability of pathogenic bacteria.These disorders activate intestinal TLRs pattern recognition receptor protein,cause further imbalance of immune and inflammatory response,and aggravate pneumonia.The intestinal flora disturbance of gastrointestinal fever complicated with pneumonia can also affect the material energy metabolism and the level of inflammatory factors.The mechanism may play a role in affecting the metabolism and energy supply of the body. |
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