| Objective:1.Based on computer algorithm image analysis technology,to study the characteristics of tongue distribution in patients with idiopathic membranous nephropathy(IMN),and the relationship between tongue characteristics and patients’ clinical physicochemical indicators,with a view to improving the standardization and objectivity of clinical research on IMN tongue diagnosis.2.Based on 16S rRNA full-length sequencing technology,to analyse the similarities and differences between the tongue my cobiota of IMN patients and healthy individuals,as well as the differences in the diversity,species composition and community composition of the tongue my cobiota of IMN patients with yellow and white greasy tongue coating,and to investigate the characteristics of the tongue mycobiota of IMN patients and IMN patients with white and yellow greasy tongue coating.Methods:Study 1:Using an epidemiological single-centre,cross-sectional survey study method,a clinical case report questionnaire was developed.A total of 105 patients with IMN who attended the outpatient clinic and ward of the Department of Nephrology,Xiyuan Hospital,Chinese Academy of Traditional Chinese Medicine were recruited from October 2021 to December 2022.A computer algorithm analysis technique "2D/3D tongue analysis method and system in open environment" was applied to qualitative and quantitative analysis of the patients’ tongue pictures.SPSS 26.0 was used to statistically describe the clinical data and objective tongue image data of IMN,further analyze the correlation between the relevant physical and chemical indicators and tongue image characteristics,and use binary logistic regression analysis to analyze the factors influencing the tongue image characteristics of IMN patients in risk stratification.Study 2:An exploratory study of tongue microbiota in patients with IMN.Twenty-four patients with IMN from Study 1 were enrolled,including 11 with white and 13 with yellowish mosses,and 10 healthy controls were recruited.Tongue flora samples were collected from these patients and sequenced using 16S rRNA full-length sequencing technology,followed by species annotation of the sequencing results and biological information analysis of Alpha diversity,Beta diversity,species compositions and compositional differences among the microbial communities of the tongue samples.Results:Study 1:1.A total of 105 patients with IMN were included in this study,of whom 62 were male and 43 were female.The mean age was in the range of 54.73±14.32 years,with middle-aged people being the most numerous at 60%,followed by elderly people at 21.0%,and young people less frequently at 19.0%.Height was 1.68±0.08m,weight was 70.93±11.26kg and BMI was 24.99±3.18kg/m2.2.The pathological distribution was 1-5 years in 61 cases(58.10%),5-10 years in 17 cases(16.19%),less than 1 year in 16 cases(15.24%)and more than 10 years in 11 cases(10.48%).Pathological staging was 30 cases(28.57%)in stage I,31 cases(29.52%)in stage II,16 cases(15.48%)in stage Ⅰ-Ⅱ and 10 cases(15.24%)in stage Ⅱ-Ⅲ;another 25 patients(23.81%)with IMN associated with positive anti-PLA2R antibodies were collected.65 cases(61.9%)in stage CKD1,24 cases(22.86%)in stage CKD2 and 22.86%),10 cases(9.52%)in CKD stage 3 and 6 cases(5.71%)in CKD stage 4.There were 64 cases(60.95%)at low risk,22 cases(20.95%)at high risk and 19 cases(18.10%)at intermediate risk.3.Characteristics of tongue texture distribution in IMN patients:(ⅰ)Tongue colour:pale dull(dark)tongue in 53 cases(50.48%),pale red tongue in 28 cases(26.67%),pale(white)tongue in 10 cases(9.52%),red tongue in 9 cases(8.57%),cyanotic tongue in 4 cases(3.81%)and vivid red tongue in 1 case(0.95%).(ⅱ)Fat or thin tongue shape:48 cases(45.71%)with normal tongue shape,47 cases(44.76%)with fat tongue and 10 cases(9.52%)with thin tongue.There was a difference in tongue shape between the different risk strata of IMN patients(P=0.049)and a further two-by-two comparison showed that a significantly higher proportion of patients with low risk of IMN had a fat tongue(p=0.01).(ⅲ)Presence or absence of dentition:79(75%)dentate tongues and 26(24.76%)non-dentate tongues.(ⅳ)With or without fissures:75 cases(71.43%)with fissured tongue and 30 cases(29%)with non-fissured tongue.4.Characteristics of tongue coating distribution in IMN patients:(ⅰ)Tongue coating colour:white coating in 77 cases(73.77%),pale yellow coating in 23 cases(22.86%)and yellow coating in 4 cases(4.84%).There was a statistical difference in the distribution of tongue colour between the different risk strata(P=0.017)and a two-by-two comparison analysis revealed a significantly higher proportion of white moss in patients with low risk of IMN(P=0.003).(ii)Thin and thick coating:67 cases(63.81%)with thin coating and 38 cases(36.19%)with thick coating.(ⅲ)Greasy and putrid coating:63 cases(60.00%)with greasy coating,40 cases(38.1%)with non-greasy and non-putrid coating,and 2 cases(1.90%)with putrid coating.(ⅳ)Integration of tongue elements to determine characteristics:The various elements of the tongue,such as moss colour,coating thickness and coating putridity,were integrated to produce seven types of tongue,namely thin white coating,thin yellow coating,thick white coating,thick yellow coating,thin white coating,thin yellow coating,thick white coating,thick yellow coating,thick white coating,thick white coating and thick yellow coating,of which 39 cases(37.14%)were thin white coating,24 cases(22.86%)were thin white coating and 21 cases(20.90%)were thick yellow coating.There was a difference in the distribution of tongue integration features between the different risk strata of IMN patients(P=0.009),with a significantly higher proportion of patients at low risk of IMN with thin white and thick white putty than at medium to high risk,and a significantly higher proportion at medium to high risk with thick yellow putty.The proportion of thin white and white thick greasy coating was significantly higher in patients at low risk of IMN than at medium to high risk,while the proportion of yellow thick greasy coating was significantly higher in patients at medium to high risk.5.Analysis of differences in physicochemical indicators between IMN patients with different tongue characteristics:24h-UTP was significantly different between IMN patients with fat and thin tongues(P=0.032),IMN patients with thin tongues had higher 24h-UTP levels than IMN patients with normal and fat tongues.24h-UTP,TP,ALB and TG levels were significantly different between IMN patients with white and yellow tongues,IMN patients with yellow tongues had significantly higher 24h-UTP and TG levels than IMN patients with white tongues(P=0.009,P=0.031),while TP and ALB levels were significantly lower than IMN patients with white tongues(P=0.009,P=0.031).Patients with IMN yellow moss had significantly higher 24h-UTP and TG levels than patients with IMN white moss(P=0.009,P=0.031),while TP and ALB levels were significantly lower than patients with IMN white moss(P=0.006,P=0.009).Patients with IMN thick coating had significantly higher 24h-UTP levels than patients with IMN thin coating(P=0.042),while ALB levels were significantly lower than patients with thin coating(P=0.030).There were no significant differences in clinical physicochemical indicators between the distribution of tongue colour,the presence or absence of denticles and fissures in the distribution of the tongue and the distribution of greasy and putrid moss in other IMN patients(P>0.05).6.Correlation analysis between tongue features and physicochemical indicators in IMN patients:coating colour in IMN patients was significantly positively correlated with 24h-UTP(R=0.263,P=0.007)and TG(R=0.242,P=0.013)and negatively correlated with TP(R=0.276,P=0.004)and ALB(R=0.231,P=0.018).Mean tongue redness in IMN patients was significantly and positively correlated with TC levels(R=0.217,P=0.026);mean tongue purpleness was significantly and negatively correlated with TP(R=0.213,P=0.018),ALB(R=0.209,P=0.032)and TC(R=0.239,P=0.014);and the area of petechiae in IMN patients was statistically negatively correlated with 24h-UTP(R=-0.203,P=0.038).7.Logistic regression analysis found that patients with fat-tongue IMN were 5.297 times more likely to progress to intermediate to high risk than patients with non-fat-tongue IMN(OR2.281,95%CI 1.166-6.812,P=0.021);patients with IMN with a mushy tongue were 4.998 times more likely to progress to intermediate to high risk than patients with non-mushy IMN(OR=3.105,95%CI 1.510-8.385,P=0.025).OR=3.105,95%CI 1.510-8.385,P=0.025),Study 2:1.A total of 24 IMN patients were included in Study 2,including 11 with white greasy moss and 13 with yellow greasy moss,and 10 healthy individuals with thin white moss were recruited externally as controls.34 tongue samples were collected,with an average of 18851.88±652.03 total CCS sequence bars detected per sample,and the number of bases measured was 15711240.07±966004.06bp.The average length of the full-length sequences obtained was 1477.62±5.14bp.2.There was no statistical difference in Alpha diversity and Beta diversity between IMN patients and healthy individuals(P>0.05),and tongue colonies of IMN patients and healthy controls had similar abundance and uniformity;IMN patients and healthy controls had similar dominant phylum,genus and species composition,and by plotting the stacked species composition at the phylum,genus and species levels,it was found that some genera and species The relative abundance of some genera and species was found to be significantly different.3.Compared the Alpha diversity and Beta diversity between the IMN yellow greasy moss group,the white greasy moss group and the healthy control tongue samples,no statistical differences were found(P>0.05),and the IMN white greasy moss,yellow greasy moss and the healthy control tongue colonies all had similar abundance and uniformity.The genera and species whose relative abundance of tongue flora in the three groups at the genus and species level were counted before 0.01 were found to be significantly different(Kruskal-Wallis,P<0.05)in two different genera,including Porphyromonas and Campylobacter,and four different species,namely Campylobacter_sp002139875,Campylobacter_fetus,Capnocytophaga_sputigena and Streptococcus_sp0001587175.with a significant decrease in the relative abundance of Porphyromonas in the white and yellow moss groups;The relative abundance of Campylobacter,Campylobacter_sp002139875 and Campylobacter_fetus was significantly higher in the yellow greasy moss group and significantly enriched;the relative abundance of Capnocytophaga_sputigena in the tongue flora of the white greasy moss group was significantly higher than the other two groups.Conclusions:Study 1:1.Patients with IMN have a predominantly pale dull(dark)tongue,with a predominantly fat tongue,mostly with teeth marks and fissures.The colour of the fur is predominantly white,and the texture of the fur is usually thin and greasy.2.Yellow,thick and greasy fur may be a potential marker for patients at high risk of IMN.3.Patients with IMN fatty tongue are 5.297 times more likely to progress to intermediate to high risk than patients with IMN non-fatty tongue;patients with IMN mushy fur are 4.998 times more likely to progress to intermediate to high risk than patients with IMN non-mushy fur.Study 2:1.Patients with IMN have a dysbiosis of the tongue flora.Porphyromonas may be the main phenotype of the tongue flora in patients with IMN.2.Campylobacter,Campylobacter_sp002139875,Campylobacter_fetus may be the hallmark organisms of IMN yellowish greasy fur.3.Capnocytophago_sputigena may be the hallmark strain of white cloying fur. |
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