Objective To evaluate the performance of drainage through micropores in the treatment of axillary osmidrosis with subcutaneous trimming.Methods Forty-eight patients with axillary osmidrosis were enrolled in this study,and treated by subcutaneous trimming.After removal of apocrine sweat glands and hair follicles,a scalpel was used to cut several micropores (measuring 3 mm in length) along dermal ridges in skin flaps.Then,drainage was carried out through the micropores.Subsequently,the incisions were sewed up with 3-0 sutures followed by compression bandaging with elastic bandages.Dressings were changed and incisions were examined at 24 hours after operation,and bandages were taken off and skin flap viability was evaluated at 10 days.All the patients were followed for 3 months.Results After treatment,sweating was decreased and axillary odor disappeared in all the patients with a return to normal social activities.No obvious scar formed.Conclusion Drainage through micropores performs well in the treatment of axillary osmidrosis with subcutaneous trimming.

Objective To determine the stage of vitiligo according to clinical and skin computed tomography (CT) features.Methods The stage of vitiligo was determined in 200 patients according to a questionnaire-based analysis of clinical features and skin CT findings respectively.Rapid progressive stage was defined as the score for vitiligous lesions or CT findings being higher than 2,slow progressive stage as the score varying from 1 to 2,and stable stage as the score less than 1.Skin samples were obtained at lesional sites receiving CT examination from 5 patients with progressive and 5 patients with stable vitiligo,and subjected to hematoxylin-eosin (HE) staining.Results No significant difference was observed between staging results based on clinical features and skin CT findings.The patients diagnosed with progressive vitiligo based on clinical features presented with grey-white or pale white patches with elevated or obscure boundaries,or trichrome vitiligo,while those with stable vitiligo presented with milky-white or porcelain-white lesions with sharp boundaries and pigmented islands.As CT showed,progressive vitiligo was characterized by a loss of integrity in dermal papillary rings at the dermo-epidermal junction level,obscure boundaries between vitiligo lesions and surrounding normal skin,presence of highly refractive cells at the dermo-epidermal junction level in the margins of vitiligo lesions,while stable vitiligo characterized by complete absence of dermal papillary rings at the dermo-epidermal junction level,sharp boundaries between the lesions and surrounding normal skin,and presence of highly refractive dendritic melanocytes.HE staining showed plenty of CD8 + T lymphocytes in the papillary dermis in the margins of progressive vitiligo lesions,but no CD8+ T lymphocyte was seen in those of stable vitiligo lesions.Conclusions Both clinical and skin CT features can be used to determine the stage of vitiligo,and the staging results are consistent with those based on histopathological findings.

Objective To evaluate the effect of puerarin on melanogenesis in melanocytes,and to explore its possible mechanisms.Methods Third-to fifth-passage melanocytes isolated from human foreskin were treated with different concentrations (1,5,10,20,40,80 and 160 μmol/L) of puerarin for 24 hours,with those receiving no treatment as the normal control group.Methyl thiazolyl tetrazolium (MTT) assay was performed to evaluate the proliferative activity of melanocytes,a sodium hydroxide solubilization method was used to measure melanin content,and reverse transcription PCR (RT-PCR) and Western blot analysis were performed to quantify the mRNA and protein expressions of microphthalmia-associated transcription factor (MITF),tyrosinase (TYR) and tyrosinase-related protein-1 (TRP-1) respectively.Results There were no significant differences in the proliferative activity of melanocytes between the puerarin (1-40 μmol/L) groups and normal control group (P ＞ 0.05),and 40 μmol/L was chosen as the concentration of puerarin for subsequent experiments.Compared with the normal control group,the 40-μmol/L puerarin group showed increased melanin content as well as mRNA and protein expressions of MITF,TYR and TRP-1 (all P ＜ 0.05).Concretely speaking,the protein expressions of MITF,TYR and TRP-1 in the 40-μmol/L puerarin group were increased by 8.69％,10.28％ and 10.58％ compared with the normal control group respectively (all P ＜ 0.05),and their mRNA expressions were 2.48,1.91 and 1.63 times higher in the 40-μmol/L puerarin group than in the normal control group respectively (all P ＜ 0.05).Conclusion Puerarin can increase the mRNA and protein expressions of MITF,TYR and TRP-1,and promote melanogenesis in melanocytes.

Objective To study the effect of epigallocatechin gallate (EGCG) and fructus psoraleae on the induction of vitiligo-like depigmentation by monobenzone in mice.Methods Forty C57BL/6 mice were included in this study.Hairs in an area measuring 2 cm × 2 cm in size were shaved on the back of each of these mice.Then,the mice were randomly and equally divided into four groups to be topically treated with vaseline cream (negative control group),monobenzone 40％ cream (model group),EGCG 5％ cream followed by monobenzone 40％ cream (EGCG group),fructus psoraleae 7％ cream followed by monobenzone 40％ cream (fructus psoraleae group),on the shaved area,respectively,for 50 consecutive days.Depigmentation of skin and hairs was observed daily by naked eyes for 15 days after drug withdrawal.At the end of the study,all the mice were sacrificed,and skin specimens were resected from the tested regions in them.Hematoxylin and eosin (HE) staining was performed to observe lymphocyte infiltration,and immunofluorescence assay to estimate the frequency of CD8 + T cells.Results Depigmentation was observed in monobenzone-induced and-uninduced sites in the model group,and in monobenzone-induced sites in all the mice in the EGCG group and fructus psoraleae group,but in neither monobenzone-induced nor-uninduced sites ih the negative control group.The average time for the appearance of depigmentation at monobenzone-induced sites was 16.7,29.3 and 19.9 days in the model group,EGCG group and fructus psoraleae group respectively.The depigmentation area index at monobenzone-induced sites was 4.00 ± 0.00 in the model group,significantly different from that in the EGCG group and fructus psoraleae group (2.11 ± 0.54 and 2.84 ± 0.79,both P ＜ 0.05).Significant differences were also observed in depigmentation area index at monobenzone-induced sites among the model group,EGCG group and fructus psoraleae group (F =14.173,P ＜ 0.05),and at monobenzone-uninduced sites between fructus psoraleae group and EGCG group (P ＜ 0.05).The frequency (expressed as fluorescence intensity) of CD8+ T cells was significantly lower in the EGCG group and fructus psoraleae group than in the model group,and significantly different between EGCG group and fructus psoraleae group (P ＜ 0.05).Conclusions Both EGCG and fructus psoraleae,especially EGCG,can interfere with the induction of vitiligo-like depigmentation of skin and hairs by monobenzone in mice.The mouse model of vitiligolike depigmentaion in this study shows higher similarity to human vitiligo.

Objective To compare the therapeutic effect of topical application of tea polyphenol versus pimecrolimus versus tacrolimus for monobenzone-induced vitiligo-like depigmentation in mice.Methods Twentyfive 3-week-old female C57BL/6 mice were randomly and equally divided into 5 groups:negative control group,model group,tea polyphenol group,pimecrolimus group,tacrolimus group.Monobenzone 45％ cream was applied to the back of mice in all the five groups except the negative control group once daily for 40 consecutive days to establish a model of vitiligo-like depigmentation.During the induction of depigmentation,the tea polyphenol group,pimecrolimus group and tacrolimus group were topically treated with tea polyphenol,pimecrolimus and tacrolimus respectively,and the model group remaining untreated.The depigmentation of hairs and skin was observed by naked eyes on a daily basis.Tissue specimens were obtained for histological examination from depigmented skin at nonapplication sites in mice after the end of the experiment.Hematoxylin-eosin staining was conducted to analyze lymphocytic infiltration,reflectance confocal microscopy to observe melanin and melanocytes in skin,and immunofluorescence assay to detect CD8+ T cell infiltration.Results Depigmentation occurred in both application sites and non-application sites of mice in the model group.Compared with the model group,the tacrolimus,pimecrolimus and tea polyphenol groups showed delayed depigmentation,reduced degree and area index of depigmentation,and attenuated lymphocytic infiltration and CD8 + T cell infiltration in depigmented maculae at application sites.In addition,the therapeutic effect of tacrolimus was stronger than that of pimecrolimus and tea polyphenol.Conclusion Tea polyphenol,pimecrolimus and tacrolimus are all effective for the treatment of vitiligolike depigmentation in mice.

Objective To evaluate imaging features of verruca plana by dermoscopy and reflectance confocal microscopy (RCM).Methods Totally,88 untreated patients with verruca plana were enrolled into this study.One typical skin lesion was chosen from each patient and subjected to dermoscopy and RCM separately,and imaging features were recorded and analyzed.Results Dermoscopy revealed a light red background in 48 cases (54.55％),a light yellow background in 40 cases (45.45％),and a various number of punctate hemorrhages in 84 cases (95.45％).RCM showed concentric onion-skin-like structures in 48 patients (54.55％).Among 57 patients with disease duration ＞ 1 year,44 (77.19％) had the concentric onion-skin-like pattern,while only 4 (12.90％) patients had similar structures among 31 patients with disease duration ＜ 1 year.A significant difference in the prevalence of concentric onion-skin-like structures was observed between patients with disease duration ＞ 1 year and those with disease duration ＜ 1 year (x2 =33.47,P ＜ 0.05).Of 88 patients,86 (97.73％) had hyperpigmentation in the basal layer at the dermo-epidermal junction,intact dermal papillary rings,vasodilatation,hyperemia and 1-2 dilated capillaries in the dermal papillae.Eight patients were treated for 2 weeks,and the above features disappeared after the treatment under dermoscopy and RCM.Conclusion Based on clinical manifestations,dermoscopy and RCM features may be helpful for the diagnosis of verruca plana.

Objective To investigate the distribution and epidemiology of fungal pathogens in zoonotic dermatophytoses.Methods Seventy-four patients with dermatophytoses and 72 pets from 64 families,who were all culture positive for dermatophytes,were included in this study and classified into 64 family-based groups.Fungal culture and direct microscopic examination were carried out for species identification of fungal isolates,internal transcribed spacer (ITS) sequence analysis and random amplified polymorphic DNA (RAPD) analysis were performed for molecular identification and homology analysis.Results Dermatophyte species were consistent among the patients and pets from the same families for all the 64 family-based groups.A total of 146 fungal strains were isolated,including 93 Microsporum canis (M.canis) strains and 53 Trichophyton interdigitale (T.interdigitale) strains.M.canis was isolated from 42 (65.7％) family-based groups including 34 groups keeping cats and 8 groups keeping dogs,while T.interdigitale from 22 (34.3％) groups,including 14 groups keeping rabbits,6 groups keeping cats and 2 groups keeping dogs.There were 54 (75.0％) pets with obvious clinical symptoms (erythema,desquamation,depilation,etc),and 18 (25.0％) asymptomatic pets which were all cats.Among the 18 asymptomatic cats,14 carried M.canis,and 4 T.interdigitale.ITS sequencing and RAPD analysis revealed a high homology between the fungal pathogens in the same family-based groups.Conclusions M.canis and T.interdigitale are common species of dermatophytes in zoonotic dermatophytoses,and both of them have host specificity.Zoonotic dermatophytes can be transmitted between human and domestic animals,and attention should be paid to asymptomatic animals (carriers).

Objective To investigate the relationship between the efficacy of autologous cultured melanocyte transplantation and serum levels of interleukin-17 (IL-17) and FoxP3 in patients with vitiligo.Methods Forty patients with stable vitiligo vulgaris were included in this study,and received autologous cultured melanocyte transplantation.Six months after the transplantation,treatment efficacy was evaluated,and patients were classified into the successfully treated group (n =25) and unsuccessfully treated group (n =15).Peripheral blood was collected from all the patients before the transplantation,and enzyme-linked immunosorbent assay (ELISA) was performed to measure the serum levels of IL-17 and FoxP3.Statistical analysis was done using two independent samples t-test with the SPSS software (version 17.0).Results The successfully treated patients showed lower serum levels of IL-17 ((15.29 ± 7.86) vs.(43.88 ± 13.02) ng/L,P ＜ 0.05),but higher serum levels of FoxP3 ((6.08 ± 2.03) vs.(3.37 ± 1.81) ng/L,P ＜ 0.05) than the unsuccessfully treated patients.Conclusion The increased serum IL-17 and decreased serum FoxP3 may contribute to the failure of autologous cultured melanocyte transplantation in patients with vitiligo.

Objective To investigate the protective effect of acetylated epigallocatechin gallate (AcEGCG) against H2O2-induced oxidative damage to human epidermal melanocytes,and to explore its possible mechanism.Methods Human epidermal melanocytes were isolated and cultured in vitro.Some melanocytes were classified into a H2O2 group induced by H2O2 only,EGCG groups and AcEGCG groups induced by H2O2 after pretreatment with different concentrations of EGCG and AcEGCG,respectively.Three concentrations (10,20 and 40 μmol/L) of EGCG or AcEGCG were used to treat melanocytes for 1 hour in MTS assay and lactate dehydrogenase (LDH) leakage assay and for 2 hours in Western blot assay,while only one concentration (40 μmol/L) was used to treat melanocytes for 0.5,1,2 and 4 hours respectively in flow cytometry assay.Some melanocytes treated with only culture medium and 0.1％ dimethyl sulphoxide (DMSO) served as the control group.After additional culture,MTS assay was performed to determine cell survival rate,flow cytometry to detect the level of reactive oxygen species (ROS) in melanocytes,Western blot to measure the expressions of caspase-9 and caspase-3 proteins.Lactate dehydrogenase (LDH) kit was used to detect the leakage of LDH to culture medium.Statistical analysis was carried out by using one-way analysis of variance for comparisons of multiple group means followed by Student-Newman-Keuls-q (SNK-q) test for multiple comparisons.Results Compared with the control group,the H2O2 group showed significantly decreased cell survival rate (22.99％ ± 0.53％,P ＜ 0.01),but increased LDH leakage level (36.58％ ± 0.73％,P ＜ 0.01),intracellular ROS level (19.08 ± 0.57,P ＜ 0.01),as well as caspase-9 (2.65 ± 0.079,P ＜ 0.01) and caspase-3 (2.36 ± 0.057,P ＜ 0.01) expressions.In comparison with the H2O2 group,the cell survival rate was significantly higher in the 10-,20-and 40-μmol/L AcEGCG groups (79.50％ ± 3.62％,86.52％ ± 5.13％,97.81％ ± 5.21％,respectively,all P＜ 0.01) and EGCG groups (43.19％ ± 1.68％,63.34％ ± 3.60％,70.82％ ± 2.1％,respectively,all P ＜ 0.01).However,the 10-,20-and 40-μ mol/L AcEGCG groups and EGCG groups all showed a significant decrease in the expression levels of caspase-9 (AcEGCG groups:1.44 ± 0.067,1.26 ± 0.059 and 1.10 ± 0.072 respectively;EGCG groups:2.31 ± 0.085,2.13 ± 0.091 and 1.35 ± 0.064 respectively,all P ＜ 0.05) and caspase-3 (AcEGCG groups:1.70 ± 0.053,1.57 ± 0.057 and 1.24 t 0.068 respectively,all P＜ 0.05;EGCG groups:2.09 ± 0.076,1.98 ± 0.093 and 1.79 ± 0.056 respectively,all P ＜ 0.05) compared with the H2O2 group.Similarly,a significant reduction was observed in the leakage level of LDH in these AcEGCG and EGCG groups (all P ＜ 0.01) and in ROS levels in the 40-μmol/L AcEGCG and EGCG groups when compared with the H2O2 group.Conclusions AcEGCG has a stronger protective effect against H2O2-induced oxidative damage to human epidermal melanocytes compared with EGCG,which may be realized through clearance of free radicals,antioxidant effects,and decrease of caspase-9 and caspase-3 expressions.

Objective To evaluate the efficacy of autologous cultured melanocyte transplantation in the treatment of genital vitiligo.Methods A retrospective analysis was carried out.From 2013 to 2016,6 patients with genital vitiligo were enrolled from Department of Dermatology,Hangzhou Third People's Hospital,who received the treatment with autologous cultured melanocyte transplantation.These patients aged 13-29 years,and the age of onset and disease duration ranged from 10 to 19 years and 2 to 10 years respectively.The area of white patches at the genital site ranged from 10 to 25 cm2.Before the transplantation,all the white patches of the 6 patients bad been stable for ＞ 12 months.Two weeks after the treatment,pimecrolimus was applied to the wounds twice a day after crust shedding.When the patient achieved ＞ 90％ repigmentation of the white patches,pimecrolimus was applied once per three days for maintenance treatment.After the treatment,these patients were followed up for at least 24 months,and the longest follow-up period was 60 months.Results Obvious pigmentation was observed in all the 6 patients 2 months after the transplantation.Meanwhile,3 patients achieved 80％ repigmentation,2 achieved 70％ repigmentation,and 1 achieved 50％ repigmentation.Six months after the treatment,3 patients achieved 95％ repigmentation,2 achieved 80％ repigmentation,and 1 still maintained 50％ repigmentation.Twelve months after the treatment,95％ repigmentation was still maintained in the 3 patients,90％ repigmentation was achieved in 2 patients,and 1 patient maintained 50％ repigmentation,and these outcomes lasted until the end of follow-up.During the follow-up,no adverse reactions were observed.Conclusion Autologous cultured melanocyte transplantation is effective for the treatment of stable genital vitiligo.