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Objective:To analyze the skin microbiota diversity in patients with pemphigus vulgaris (PV) using 16S rDNA sequencing.Methods:Ten patients with PV and 10 healthy controls were collected from the Department of Dermatology, the Third People′s Hospital of Hangzhou. Skin swabs were collected from perilesional skin (PV group) and contralateral non-lesional skin (PVn group) of the patients with PV, as well as from the corresponding body sites of the healthy controls (normal control group) . The 16S rDNA amplicon sequencing technology was used for gene sequencing and classification in all microbiota samples, and Usearch software for data cluster analysis to obtain operational taxonomic units (OTUs) and assess species abundance at the phylum, class, order, family and genus levels. Observed species index, Shannon index and Simpson index were used to estimate α diversity, and principal coordinate analysis (PCoA) was performed to analyze β diversity. Linear discriminant analysis effect size (LEfSe) analysis was conducted to identify differentially abundant species in each group. PICRUSt software was used for gene function prediction. Wilcoxon rank sum test was used as nonparametric test for comparisons between 2 groups, and Kruskal Waills test as nonparametric test for comparisons among 3 groups.Results:There were 2 002, 1 869, 1 751, 1 611 and 1 120 OTUs at phylum, class, order, family and genus levels respectively. Cluster analysis showed that skin microbiome in the 3 groups mainly consisted of Firmicutes, Actinobacteria, Bacteroidetes and Proteobacteria at the phylum level. At the genus level, Staphylococcus was the most abundant in the PV group and PVn group, and Corynebacterium was the most abundant in the normal control group. The observed species index, Shannon index and Simpson index all significantly differed among the 3 groups (all P< 0.05) , and the Shannon index and Simpson index were significantly lower in the PV group (3.24±1.30, 0.70±0.19, respectively) than in the normal control group ( P< 0.05) . PCoA analysis showed no significant difference in β diversity among the 3 groups ( P=0.054) . Rank sum test showed that the abundance of 32 species significantly differed among the 3 groups ( P< 0.05) . Among them, high relative abundance was observed in the class Bacilli enriched in the PV group, as well as the genera Micrococcus and Brevundimonas enriched in the normal control group. According to the disease duration, the patients with PV were divided into long-course PV group with disease duration of ≥ 3 months, and short-course PV group with disease duration of < 3 months. Clostridiales, Oscillibacter, Sphingomonas were enriched in the long-course PV group, and Gammaproteobacteria was enriched in the short-course PV group. Gene function prediction analysis showed that the genes related to infectious diseases were enriched in the pemphigus group. Conclusion:The 16S rDNA-based microbiota profiling suggested differences in the diversity and composition of skin microbiota between patients with PV and healthy individuals.
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Objective To identify novel biomarkers from urinary protein profiles for the early diagnosis of nephritis in patients with Henoch-Sch(o)nlein purpura by surface enhanced laser desorption/ionization time of flight-mass spectrometry (SELDI-TOF-MS) technique.Methods Urine samples were collected from 60 untreated patients with Henoch-Schonlein purpura,including 30 patients with nephritis and 30 without.SELDI-TOF-MS technique was used to characterize the protein profile in these urine samples,and the Zhejiang University Cancer Institute-Protein Chip Data Analysis System (ZUCI-PDAS) to identify urine protein markers and construct diagnostic model for nephritis in patients with Henoch-Schonlein purpura.Results Totally,154mass peaks were identified with high quality,and two proteins at a mass-to-charge ratio (m/z) of 2454.971 and 2439.686 showed significantly differential expression between the two groups of patients (P ＜ 0.05).Seven biomarkers were used to establish a diagnostic model.As estimated by the leave-one-out cross-validation,the diagnostic model distinguished patients with nephritis from those without with a specificity of 71％ and sensitivity of 84％.Conclusions The developed diagnostic model based on SELDI-TOF-MS technique and bioinformatics is somewhat specific and sensitive for the prediction of nephritis in patients with Henoch-Schonlein purpura.
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Objective: To screen and identify gene mutations of 11 Chinese patients with Hailey Hailey disease (HHD). Methods: Cases of HHD were diagnosed by history, clinical menifestations and pathology. Then genomic DNA samples of patients were extracted from perpheral blood leukocytes, and polymerase chain reaction(PCR), DNA sequencing were performed. Results: We found five mutations in ATP2C1 gene including 3 nonsense mutations and 2 splicing mutations. Four of them were novel mutations. Conclusion: Both nonsense mutation and splicing mutation could affect the rusult of transcription,translation, and the functions of protein encoded by ATP2C1 gene, so the mutations reported in this study is the underlying cause of HHD.
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Objective To investigate family history, sex, age of onset, disease severity and environmental predisposing factors in vitiligo patients. Methods Eight hundred and fifteen vitiligo patients were investigated by questionnaires. Patients with family history were compared with those without such history. SPSS 10.0 software package was applied for data analysis. Results Of 815 vitiligo probands, 128 had family history and 687 did not. The heritability rate was 15.7%. Compared with general population, vitiligo probands with affected fathers or mothers had a relative risk (RR) of 132 or 72, respectively. The RRs of those with affected first-degree relatives varied from 12 to 28. There was no significant difference of mean age of onset and disease severity between patients of paternal inheritance and maternal inheritance. No significant difference was found regarding sex and mean age of onset between the groups with and without family history. However, the patients with family history were more likely to have scattered, bilateral distributed and progressive vitiligo. Of the environmental predisposing factors, the mean daily sun-exposure time was closely related to vitiligo in patients with family history. Conclusion Genetic factors may play an important role in the pathogenesis and disease severity of vitiligo.
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Objective To identify gene mutations of two cases of epidermolytic hyperkeratosis ichthyosis.Methods Punch biopsies were taken from typical lesions for histopathological examination by light microscopy and transmission electron microscopy.Genomic DNA was extracted from blood samples.Mutations of keratin1(K1)and keratin10(K10)were detected by polymerase chain reaction(PCR)and DNA sequencing;Frequencies of the alleles were screened by PCR-based allele-specific assays(PASA)and restriction fragment-length polymorphism(RFLP)in normal controls.Results There was a single heterozygous point mutation in either K1or K10genes,i.e.2140G→A of K10gene and4226G→A of K1gene,leading to an amino acid alteration of arginine to histidine(K10R156H)and glutamic acid to lysine(K1E477K),respectively.These substitutions were not found in normal controls.Conclusion K10R156H and K1E477K mutations were the cause of the phenotypes in these two cases.