| ObjectiveUsing multivariate statistical methods to explore the syndrome pattern of acute cerebral hemorrhage.On this basis,the characteristics of the flora of acute cerebral hemorrhage of liver-yang sputum were analyzed,which provided a micro-level basis for the study of TCM syndromes.MethodsThe first part of the literature study:By consulting various literatures,the latest status of diagnosis and treatment of acute cerebral hemorrhage in traditional Chinese and western medicine,as well as the latest research progress of intestinal flora and acute cerebral hemorrhage were understood.The second part of the retrospective study:Patients admitted to the first affiliated hospital of guangzhou university of traditional Chinese medicine due to acute cerebral hemorrhage from May 2013 to May 2018 were selected,involving 1,072 cases,and a total of 683 cases meeting the research criteria were included.Through the method of two-person and two-machine input,the four-diagnosis information of all the cases collected was input into the Excel 2010 to establish the database in the form of data.Frequency analysis,factor analysis,cluster analysis and other multivariate statistical methods were used to study the syndrome rules of acute cerebral hemorrhage.The third part of the intestinal flora research:On the basis of the second part.Fecal samples of 9 patients with acute cerebral hemorrhage due to hyperactive liver Yang type admitted to the first affiliated hospital of guangzhou university of Chinese medicine in the year of 2018 were selected as the observation group.Meanwhile,fecal samples of’ 6 healthy people who underwent physical examination were selected as the control group.Then,the total bacterial DNA was extracted from the two groups of samples.Primers were designed according to the V4 region of 16S rRNA for amplification,and Paired-End sequencing was conducted by Illumina MiSeq platform.The sequencing results were analyzed by using bioinformation analysis software,and the composition and structure differences of the two groups of samples were compared.ResultsThe first part of the literature research:The diagnosis and treatment technology of traditional Chinese and western medicine has some limitations in the diagnosis and treatment of acute cerebral hemorrhage.The research on syndrome law lacks unity.As a part of human body,intestinal flora is involved in various physiological and pathological processes of human body,which is closely related to the risk factors of acute cerebral hemorrhage.The second part of the retrospective study:Age and gender:the average age of onset of acute cerebral hemorrhage is 57.34±14.967 years old,the age of high incidence is 40-50 years old,the proportion of men and women is 3:2,the incidence rate of males is higher than that of females.On the season:the solar terms with high disease are in turn:Dahan,Dongzhi,Hanlu,Daxue,Lichun,jingzhe.winter and spring are high seasons,especially winter.Frequency analysis:A total of 683 cases were included in the study.There were 210 symptom types and signs,and the cumulative frequency was 8357 times.The pulse line was 533 times(6.4%),the acute face was 344 times(4.1%),and the limb weakness was 300 times(3.6%).Vomiting 275 times(3.3%),headache 253 times(3.1%),mental condition 246 times(3%),speech disadvantage 241 times(3%),dizziness 237 times(2.8%),passive position 234 times(2.8%),the expression pain 229(2.8%)is dominant.the frequency of the first 39 categories with higher frequency(>1%)is 6659 times,accounting for 79.7%of the total classification.Cluster analysis and factor analysis:The symptoms and signs with higher frequency(>1%)were selected for further analysis.The KMO test statistic was 0.609,and the spherical Bartlett test:χ2=5622.131,P=0.000,satisfying the factor analysis conditions,and 14 common factors were obtained.For the five cluster syndromes,the order was liver yang sputum type,qi stagnation and blood stasis type,phlegm heat phlegm type,yin deficiency yang sputum type,turbid block type.The third part of the intestinal flora research:OTU Venn graph analysis and Core-Pan OTU analysis showed that there were significant differences between the observation group and the control group in the total number of OTUs and the total number of unique OTUs.PCA analysis and PLS-DA analysis showed significant differences in composition between the observation group and the control group.The OTU Rank curve showed that the species richness and species uniformity of the observation group were lower than those of the control group.In the diversity analysis of Alph,observed species index,Chao index,ACE index,Shannon index and Simpson index of the observation group and the control group were not statistically significant(P>0.05).The dilution curve indicated that the quality control of sequencing was sufficient to reflect the species diversity of the sample.In the Beta diversity analysis,the Beta diversity matrix heat map and similarity clustering tree showed that the samples in each group had good clustering,indicating that the samples in the group had high similarity,while the main coordinate analysis showed that the two groups were significantly separated,and the two groups had significant differences in the structure.The phylogenetic tree of species shows a total of 12 phyla,121 genus,of which the proportion of Firmicutes is the highest.In the analysis of species and abundance,the ratio of the Firmicutes and the Bacteroides gate in the observation group at the gate level(F/B)increased significantly,and the Verrucomicrobia abundance of the observation group was significantly lower than that of the control group.Statistically significant(P<0.05).In the observation group,the abundance of Bacteroides,Porphyr-omonadaceae,and Erysipelotrichaceae in the observation group was higher than that in the control group,while in the control group,the Prevotellaceae and Verrucomicrobiaceae had higher richness than the observation group.In the observation group,the differences were statistically significant(P<0.05).At the genus level,the Bacteroides and Acidaminococcus of the observation group were relatively high,while the Prevotella,Akkermansia,and Blautia in the control group had higher abundance,and the differences were statistically significant.(P<0.05),and the relative abundance ratio(B/P)of Bacteroides and Prevotella in the observation group was significantly higher than that in the control group.At the Species level,the relative abundance of Bact-eroides ovatus and Bacteroides fragilis in the observation group was higher,while the relative abundance of Prevotella copri,Akkermansia muciniphila,and Ruminococcus callidus in the control group was higher,and the difference was statistically significant(P<0.05).In the LEFSE analysis,abnormalities of Bacteroides,Prevotella,and Verrucomicrobiaceae were closely related to acute cerebral hemorrhage of liver-yang hyperactivity.Conclusion1.Cluster analysis and factor analysis:TCM syndromes of acute cerebral hemorrhage are mainly composed of liver-yang sputum type,qi stagnation and blood stasis type,phlegm-heat phlegm type,phlegm block type,and yin deficiency and yang type.Among them,liver yang is the most common type.2.The changes in the composition and diversity of intestinal flora are important causes of acute cerebral hemorrhage in the liver and yang,and The relative abundance ratio(F/B)of the Firmicutes and Bacteroides and the relative abundance ratio(B/P)of Bacteroides and Prevotella are acute cerebral hemorrhage One of the important reference indicators.3.Decreased relative abundance of Prevotella copri and Akkermansia muciniphila,and increased relative abundance of Bacteroides fragilis are important factors in acute cerebral hemorrhage. |
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