Investigation Of The Relationship Between Skin Microbiota,Skin Physical Barrier Function,and Balanoposthitis

Objective: Balanoposthitis is a common genital skin disease in men.It can be caused by a variety of reasons and embodies as a type of inflammatory changes upon the skin of the penis,prepuce,and glans penis.Skin microbiota refers to the microorganisms inhabiting the surface of the skin,including bacteria,fungi and viruses.The imbalance of skin flora can participate in the occurrence of many skin diseases,such as atopic dermatitis,psoriasis,seborrheic dermatitis,acne,folliculitis,etc.The skin physical barrier function is that resistance to the invasion of microorganisms,friction,ultraviolet rays,chemical substances,etc.The impaired physical barrier function of the skin can cause skin allergies,infections,and other diseases.However,the current microbial research on balanoposthitis mostly comes from the results of traditional microbial culture,and there is no research on related skin microbiome and skin barrier function detection.Our previous work conducted a small sample of bacterial sequencing and skin physical barrier function detection on the glans skin of patients with balanoposthitis,and found that the skin barrier function was impaired in patients,the genus Finegoldia and Prevotella were suspected as pathogenic.Accordingly,this study aims to increase the sample sites(glans penis,balanopreputial sulcus,inner prepuce,and outer prepuce)of the skin bacterial and fungal flora of patients with balanoposthitis,as well as the analysis of skin physical barrier function of the glans,inner prepuce,outer prepuce,and lateral groin.To obtain a comprehensive and systematic relationship between skin flora,skin physical barrier function,and balanoposthitis.To evaluate the therapeutic effects of Lactobacillus and clioquinol on balanoposthitis.Methods: 1.In this study,29 healthy subjects and 26 patients with non-fungal balanoposthitis were tested for skin physical barrier function(transepidermal water loss,skin hydration,and p H)of the glans penis.At the same time,the corresponding skin bacterial flora was collected by using cotton swabs,and the bacterial flora distribution and biodiversity of the skin lesions were obtained by extracting sample nucleic acid,16 s r RNA gene amplification,sequencing,clustering,comparison,and annotation.Then the patients were randomly divided into three groups,group 1 used fusidic acid(once in the morning)+ moisturizer(once in the evening),group 2 used moisturizer(once in the morning)+lactobacillus(once in the evening),and group 3 used fusidic acid(once in the morning)+Lactobacillus(once in the evening);Patients were followed up after ten days of use to evaluate the treatment effects of each group and collect skin flora and skin physical barrier function data,and compare them with the state before treatment.2.We limit the age to 18-28 to reduce the influence of other factors such as sex,smoking,drinking,etc.We recruited66 eligible healthy subjects,collected sociodemographic characteristics,and then profiled sequencing of skin bacterial flora(16s r RNA)and fungal flora(ITS)at the four sites of glans penis,balanopreputial sulcus,inner prepuce,and outer prepuce;at the same time,we perform skin physical barrier function tests on the glans penis,inner prepuce,outer prepuce,and lateral groin.3.We recruited 32 patients with balanoposthitis,collected sociodemographic characteristics,and conducted a randomized,double-blind,placebocontrolled,clinical research trial evaluating the effectiveness and safety of clioquinol in the treatment of foreskin balanitis.According to the random number table provided,the patients were divided into 2 groups,one group was treated with clioquinol experimental group,and the other group was treated with clioquinol matrix placebo group;a total of 2weeks of treatment,and then withdrawal for 2 weeks,a total of 4 weeks for this experiment.Follow-up at the time of enrollment(W0),the first week of treatment(W1),the second week of treatment(W2),and the second week of discontinuation(W4);four sites,glans penis,balanopreputial sulcus,inner prepuce,and outer prepuce of sequencing of skin bacterial flora(16s r RNA)and fungal flora(ITS);at the same time,glans penis,inner prepuce,outer prepuce,and lateral groin were tested for physical barrier function of the skin.To make a horizontal comparison with the healthy subjects in the second part,as well as the comparison between the experimental group and the placebo group and the before and after comparison of each group during the treatment.Results: 1.By comparing the physical barrier function of the skin of healthy subjects(HC,n=29)with the redundant prepuce(HCRP,n=20)and the group not exceeding the coronary sulcus(HCNP,n=9),we found that HCRP had higher TEWL and SH than HCNP’s.25/26balanoposthitis(BP)showed that the foreskin exceeded the coronary sulcus.Comparing BP and HCRP,it was found that BP had low SH and high p H.After treatment with drugs such as fusidic acid(group 1)-Lactobacillus(group 2)-combination of the two(group 3),the improvement rate of the severity score in the first group was 89.5%,in the second group was 70.2%,in the third group was 80.8%.However,the patient’s skin barrier function did not change significantly.By comparing the level of bacterial flora in healthy subjects and patients,it was found that patients had significantly more Prevotella bivia and Staphylococcus warneri,and these two bacteria were positively correlated with the severity of the disease;After treatment,it was found that Prevotella bivia was significantly reduced under the intervention of Lactobacillus,but Staphylococcus was significantly increased.2.The skin barrier function of the glans penis,inner prepuce,outer prepuce,and lateral groin of 66 healthy subjects showed that SH and TEWL of the glans and inner prepuce were affected by the degree of foreskin coverage.The more covered,the higher the SH and TEWL,the p H value of the penis skin surface was about 5.5,which was not affected by the detection site and the length of the foreskin.Ecological analysis of the flora of skin fungi and bacteria in healthy subjects showed that there was no difference in Alpha diversity at the four sites.In terms of Beta diversity,the outer prepuce was similar to the other three.There are significant differences in each locus.Staphylococcus,Corynebacterium,Prevotella,Finegoldia and Porphyromonas were more common in bacteria;Malassezia is more common in fungi.3.We recruited 32 patients with balanoposthitis.Compared with the aforementioned 66 healthy subjects,the patients had more smoking,drinking and sex life.Compared with healthy subjects,the skin barrier function of the patients’ glans penis,balanopreputial sulcus,inner prepuce,and outer prepuce showed only low SH was seen.The results of the double-blind experiment have not yet been revealed.The average improvement rate of W1 severity of group A was 51.7%,of W2 was 84.8%,and of W4 was 78.3%;the average improvement rate of W1 of group B was 66.7%,and of W2 was 90.8%,and of W4 was 86.5%.In the performance of patient fungi and bacteria before and after treatment,we selected those bacteria that were enriched in W0 and decreased in W2.According to the analysis of the bacterial and fungal flora of patients,Prevotella bivia,Streptococcus agalactiae,Haemophilus parainfluenzae,Anaerococcus vaginalis,Bacteroides coagulans and other bacteria,Candida albicans,Fusarium solani and other fungi are involved in the pathogenic process of disease.Conclusion: 1.Lactobacillus have a certain ability to regulate the structure of diseased state of balanoposthitis,combined with a better improvement rate and the anaerobic environment created by overlength foreskin,treatment using Lactobacillus is worthwhile for the clinically common type of balanoposthitis with overlength foreskin.Prevotella bivia is the pathogenic bacterium,while Staphylococcus warneri may only play an auxiliary pathogenic role.2.For the skin barrier function SH and TEWL,the glans penis and inner foreskin are affected by the length of the foreskin.The larger the coverage area,the higher the SH and TEWL.The p H value of the penis skin surface is about 5.5,which is not affected by the detection site and the length of the foreskin.The lateral foreskin is different from the other sites,and the inner foreskin and the glans penis are similar in all aspects.Staphylococcus,Corynebacterium,Prevotella,Finegoldia and Porphyromonas were more common in bacteria;Malassezia is more common in fungi.3.The incidence of balanoposthitis in the population has nothing to do with the degree of foreskin coverage,but patients with more foreskin coverage generally have more severe severity.The epidermal water content in the skin physical barrier function of the glans and inner foreskin can be used to distinguish diseases or health conditions,which is decreased in patient’s epidermal water content;the p H is not affected by the degree of foreskin coverage or the severity of the disease.According to the analysis of the bacterial and fungal flora of patients,Prevotella bivia,Streptococcus agalactiae,Haemophilus parainfluenzae,Anaerococcus vaginalis,Bacteroides coagulans and other bacteria,Candida albicans,Fusarium solani and other fungi are involved in the pathogenic process of disease.