| Target:Observe the colonization of fungi in the skin lesions of patients with atopic dermatitis(AD),and guide antifungal treatment.Method:Collected patients who were diagnosed with atopic dermatitis from the Department of Dermatology and Venereology of the Second Affiliated Hospital of Kunming Medical University from July 1,2020 to January 31,2021,who met the"Chinese Atopic Dermatitis Diagnostic Criteria".Evaluate the patient’s condition(EASI score,DLQI),improve the total IgE level inspection,take the dander of the patient’s skin lesions for fungal microscopy,culture and identification,and choose the treatment plan based on the results of the patient’s fungal microscopy.If the mycoscopic examination is negative,give the affected area topical glucocorticoid;if the mycoscopic examination is positive,then give the affected area topical antifungal treatment.After 14 days of treatment,the EASI score will be performed again to calculate the efficacy index.The data conforming to the normal distribution is represented by x ±s standard deviation,if the variance is uniform,the difference is analyzed by T test,the data of skewed distribution is represented by M(Q1,Q2),and the rank sum test is used to analyze the difference.Chi-square test was used to analyze whether the positive rate of fungal microscopy and fungal culture differed between different parts and different severity of the disease.Spearman’s rank correlation was used to analyze the correlation between total IgE,DLQI score,and EASI scoreResult:1.General situation:A total of 58 AD patients were enrolled in this study,age(years)35.29±17.64,male to female ratio 1:1.07,total IgE level(IU/ml)45.99(12.54,161.10),DLQI(points)5.00(3,8),EASI(points)4.00(1.58,7.23).Among them,41 patients had mild disease severity(EASI<7 points),17 patients had moderate disease severity(EASI 7-21 points),and 0 patients had severe disease severity(EASI>21 points).A total of 58 fungal microscopy samples were collected,of which 46 were negative and 12 were positive.A total of 52 fungal culture results were collected,30 cases were negative and 22 cases were positive.2.There was no significant difference between the age,total IgE,DLQI score,and EASI of patients with positive and negative mycoscopy before treatment(P>0.05).The positive rate of fungal microscopy in different skin lesions of AD patients was not statistically significant(P>0.05).In patients with different severity of atopic dermatitis,the positive rate of fungal microscopy in the same skin lesion area was not statistically significant(P>0.05);the positive rate of fungal microscopy in different skin lesion areas in patients with mild and moderate AD had no positive rate Statistically significant(P>0.05).3.There was no significant difference between the age,total IgE,DLQI score,and EASI of patients with positive and negative fungal cultures before treatment(P>0.05).The positive rate of fungal culture in different skin lesions of AD patients was not statistically significant(P>0.05).In mild and moderate AD patients,the positive rate of fungal culture in the same skin lesion area was not statistically significant(P>0.05);in AD patients with the same disease severity,the positive rate of fungal microscopy in different skin lesion areas was not statistically significant Academic significance(P>0.05).4.The culture method identified 10 fungi,including 3 cases of Malassezia,3 cases of Candida,and 1 case each of Rhodotorula,Aspergillus,Gliocladium,Aspergillus and Spissiomyces.5.Culture-free method for ITS sequencing of skin lesions of AD patients:240,293 sequences were obtained from 20 samples,and 155,036 sequences were obtained after noise reduction,with a length of 582-1820 bp and an average length of 1226 bp.Each sample is about 126.85 ASVs/OTUs.The number of ASVs/OTUs in the C1 group(mild,EASI<7 points,n=15)was 84-173(126.74±14.16),and the ASVs/OTUs in the C2 group(moderate,7<EASI<21,n=5)The number is 78-143(86.58±15.34).More than 50%of the flora can only identify the domain,and less than 40%of the flora can identify the family,genus,and species.As a whole,the relative abundance of Malassezia and Aspergillus at the family level is the highest;at the species level,the colonization of Malassezia is mainly restricted.6.The flora abundance and species of mild and moderate AD patients have changed;at the genus level,mild patients are colonized by Chromocleista(18.11%);moderate patients are colonized by Malassezia(34.74%)Mainly colonization.At the species level,mild patients are mainly restricted to colonization of Malassezia,and moderate patients are mainly colonized by Malassezia_arunalokei(23%).7.The relative abundance of Malassezia in patients with AD can be as high as 79%,and the diversity and abundance of the flora of the patients before and after treatment have changed.The composition and abundance of the microbial community in different parts are also different.At the family level,Malassezia and Aspergillus are the dominant bacteria in each part.At the species level,the ears,upper limbs and lower limbs all had the highest abundance of restricted Malassezia,and the face and back had the highest abundance of Malassezia_arunalokei.In addition,AD patients of different genders have different dominant bacteria.Males have the highest abundance of Malassezia_arunalokei and females have the highest abundance of restricted Malassezia.8.Alpha diversity analysis of AD patients:the abundance of bacterial communities in the mild group was higher than that in the moderate group,and the difference was statistically signifiicant(P<0.05).The diversity and uniformity of the fungal community in the mild disease severity group was higher than that in the moderate disease severity group,but the difference was not statistically significant(P>0.05).9.The age of AD patients has nothing to do with total IgE,DLQI score and disease severity.Total IgE has nothing to do with disease severity and DLQI score.There is a positive correlation between the DLQI score of AD patients and the EASI score of disease severity.The severity of the condition of male patients was higher than that of female patients,and the difference was statistically significant(P<0.05).10.A total of 10 patients were revisited in this study,with a return rate of 17.24%,of which 0 cases were markedly effective,10 cases were effective,and 0 cases were ineffective.The overall effective rate was 100%.After treatment,the EASI score was significantly lower than before treatment,and the difference was statistically significant P<0.05,that is,the two treatment programs have a significant therapeutic effect.There was no statistically significant difference in the efficacy index of different topical treatment groups,P>0.05.in conclusions1.In this study,the positive rate of fungal microscopy and culture of the skin lesions of AD patients has nothing to do with the age,gender,severity of the disease,and the location of the sample.The culture method found that the skin lesions of the patient were mainly colonized by Malassezia and Candida,and fungi such as Rhodotorula and Aspergillus were also detected.The culture-free method found that Malassezia was the dominant bacteria in the skin lesions of AD patients,and Malassezia_arunalokei was identified for the first time in the skin lesions of AD patients in China.2.In this study,the abundance and types of fungal communities in the skin lesions of AD patients changed in different severity,gender,and skin lesion areas.The abundance of restricted Malassezia in mild AD patients is the highest;the moderate Malassezia_arunalokei has the highest abundance.Male patients had the highest abundance of Malassezia_arunalokei,and female patients had the highest abundance of restricted Malassezia.Ears,upper limbs and lower limbs all had the highest abundance of Malassezia restricted,and Malassezia_arunalokei on the face and back had the highest abundance.The microbial diversity of AD patients increased first and then decreased with the increase of the severity of the disease.The richness,uniformity and diversity of the fungal community in the skin lesion area of moderate patients were lower than those of mild patients.3.In this study,the quality of life score of AD patients is related to the severity of the patient’s condition.The higher the EASI score,the more severe the clinical symptoms,and the higher the DLQI score,the worse the patient’s quality of life.The severity of the condition of male AD patients is higher than that of female patients.The total IgE level of AD patients has nothing to do with the severity of the disease.4.There was no significant difference between the efficacy of combined topical antifungal therapy in AD patients in this study and the efficacy of hormones alone. |
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