A quantitative assessment of the effects of formal sun protection education on photosensitive patients.

PURPOSE: To quantitatively assess the effect of formal sun protection education on sun exposure habits and quality of life in photosensitive patients. METHODS: Patients with chronic actinic dermatitis (CAD) or polymorphous light eruption (PLE) were randomized to either the intervention or the control group. General advice about sun protection and broad-spectrum sunscreen were provided to all participants. The intervention group was given two additional intensive sun protection instruction classes at the beginning of spring and then in summer. At baseline and 12 months, each participant completed interviews that included a questionnaire about sun protection behaviors and a modified Dermatology Life Quality Index (DLQI) questionnaire. RESULTS: In the intervention group, after the first intensive sun protection instruction, all aspects of sun exposure habits were significantly improved from baseline (P < 0.01). At study's end, there had been no significant change in sun exposure habits in the control group compared with baseline, whereas sun exposure habits in the intervention group significantly improved (P < 0.01). After two intensive sun protection training sessions, the modified DLQI significantly decreased in the intervention group compared with baseline (P < 0.001), while no change was observed in the control group. CONCLUSION: Formal sun protection education improved sun exposure and protection behaviors as well as quality of life in photosensitive patients.

Dysregulation of miRNA146a versus IRAK1 induces IL-17 persistence in the psoriatic skin lesions.

Psoriasis is a common chronic inflammatory skin disorder with dysregulation of miRNAs. The expression pattern of miR-146a and target gene IRAK1 in lesions and PBMCs of plaque psoriasis remains unclear. In our study, we found the expression of miR-146a was up-regulated both in lesions and PBMCs of psoriatic patients, and positively correlated with IL-17 expression, whereas the target gene IRAK1 expression was expressed differentially in lesions and peripheral blood. Inability of miR-146a inhibiting target gene IRAK1 may contribute to the persistent inflammation in lesions of psoriasis.

Prevalence of childhood atopic dermatitis: an urban and rural community-based study in Shanghai, China.

BACKGROUND: Atopic dermatitis (AD) is a common inflammatory and chronically relapsing disorder with increasing prevalence. However, little is known about its prevalence in Shanghai, the top metropolitan of China. This study will estimate and compare the prevalence of AD in urban and rural areas in representative samples of 3 to 6-year-old children in Shanghai. METHODOLOGY/PRINCIPAL FINDINGS: A descriptive cross-sectional study was performed. Pre-school children were obtained by cluster sampling from 8 communities in different districts in Shanghai. The main instrument was the core questionnaire module for AD used in the U.K. Working Party's study. All the data were statistically analyzed by EpiData 3.1 and SPSS16.0. A total of 10,436 children completed the study satisfactorily, with a response rate of 95.8%. The prevalence of AD in 3 to 6-year-old children was 8.3% (Male: 8.5%, Female: 8.2%). The prevalence in urban areas of Shanghai was gradiently and significantly higher than that in rural areas. The highest prevalence was in the core urban area (10.2% in Xuhui Tianping) vs. the lowest far from the urban areas (4.6% in Chongming Baozhen). CONCLUSIONS/SIGNIFICANCE: The prevalence of AD was 8.3% (95%CI: 7.6%-9.1%) in children aged 3 to 6 in Shanghai. The prevalence of AD decreased from the center to the rural areas in Shanghai.

Oxidation levels differentially impact melanocytes: low versus high concentration of hydrogen peroxide promotes melanin synthesis and melanosome transfer.

BACKGROUND: UVB light can generate potentially harmful hydrogen peroxide (H(2)O(2)) in vivo, but it can also promote the beneficial proliferation and migration of melanocytes. The successful use of UVB monotherapy for treatment of vitiligo suggests that H(2)O(2) may have a biphasic effect on melanin synthesis and melanosome transfer. OBJECTIVE: To study the beneficial role of H(2)O(2) on melanogenesis and melanosome transport in living melanocytes and keratinocytes. METHODS: A co-culture system model was constructed using the primary human melanocytes and keratinocytes. The MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay was used to determine cell proliferation, NaOH was used to determine the melanin content, and real-time PCR was used to determine tyrosinase expression. Western blot was used to determine Rab-27A and protease-activated receptor 2 (PAR-2) expression. RESULTS: This study demonstrated that tyrosinase was activated by low concentrations of H(2)O(2) (≤0.3 mM); however, this activity was downregulated by high concentrations of H(2)O(2) (>0.3 mM). Activation of high levels of melanin synthesis was induced when cells were treated with low concentrations of H(2)O(2) (0.3 mM). Further observation using an in vitro co-culture system of fluorescein (carboxyfluorescein diacetate succinimidyl ester, CFDA-SE)-labeled melanocytes and keratinocytes indicated that melanosome transfer occurred in normal human epidermal melanocytes. Fluorescence microscopy revealed increased melanosome transfer into keratinocytes treated with 0.3 mM H(2)O(2) in the co-culture compared to the control. Examination of melanosomes in the keratinocytes by flow cytometry confirmed these results. Furthermore, treatment with H(2)O(2) (0.3 mM) upregulated the expression of Rab-27A and PAR-2, significant proteins involved in melanosome transfer, according to Western blot. CONCLUSION: These results confirmed that low concentration levels of H(2)O(2) play a major role in the regulation of human pigmentation by increasing melanin synthesis and melanosome transfer.

Reversal of iC3b-inhibited dendritic cell differentiation via inhibition of the extracellular signal-regulated mitogen-activated protein kinase promotes CD4(+) T cell proliferation.

OBJECTIVES: To investigate the roles of ERK1/2 and p38 MAPK cascades in the differentiation of iC3b-combined CD14(+) monocyte into CD1a(+) MDDC, and to study how these cells influence CD4(+) T cell proliferation. METHODS: CD14(+) monocyte was co-cultured with iC3b with or without inhibitors specific for ERK1/2 or p38 MAPK pathways for 2days, then the expressions of CD14, CD1a, phophso-ERK1/2, phophso-p38, IL-10 and IL-12 p70 were detected, and CD4(+) T cell proliferation was measured via (3)H-TdR as well. RESULTS: Maturation of CD1a(+) DC was inhibited by iC3b along with downregulated expressions of CD1a, phophso-p38 and IL-12p70 and upregulated expressions of phophso-ERK1/2 and IL-10, and the CD4(+) T cell proliferation was restrained accordingly. When pretreated with inhibitor specific for ERK1/2 pathway, the inhibited maturation of imDC was reversed prominently with a higher level expression of CD1a and IL-12p70, whereas expressions of phophso-ERK1/2 and IL-10 were lowered, and accordingly the CD4(+) T cell proliferation restored significantly. CONCLUSIONS: iC3b inhibited the differentiation of CD14(+) monocytes into CD1a(+) MDDCs via ERK1/2 pathway, and restoration of CD1a(+) MDDCs maturation occurred with the treatment of inhibitors specific for ERK1/2 pathway. Meanwhile, treatment of the inhibitor for the ERK1/2 cascade reversed the inhibited CD4(+) T cell proliferation, implying a potential possibility for clinical intervention.

Childhood vitiligo in China: clinical profiles and immunological findings in 620 cases.

BACKGROUND: Childhood vitiligo is a common pediatric skin disorder. The pathogenesis of vitiligo is unclear, and immunological dysfunction may play an important role. OBJECTIVES: This prospective study aimed to profile childhood vitiligo and to discuss its correlation with immunological dysfunction. METHODS: All of the 620 enrolled patients were aged younger than 14 years, and were assessed with a standard questionnaire. The levels of immunoglobulins, complement, and T-lymphocyte subsets were measured in 270 of these 620 patients. RESULTS: Of the 620 children, 302 (48.71%) were boys and 318 (51.29%) were girls, with an average disease onset age of 7.57 years. The average duration was 13.45 months. 453 (73.06%) children had head and neck involvement and 160 (25.81%) children had segmental vitiligo. 84 (13.55%) children had a family history. There was a correlation between the disease and seasons. The onset or progression usually occurred in summer and spring. Halo nevus was seen in both segmental and non-segmental vitiligo. Precipitating factors such as stress appeared more commonly in segmental vitiligo. As to the immunological findings, in segmental vitiligo, the levels of C3 and C4 were lower in the active relative to the quiescent stage (p < 0.05); and in non-segmental vitiligo, the percentages of CD3+ and CD4+ lymphocytes and the CD4+/CD8+ ratio were lower in the active relative to the quiescent stage (p < 0.01). CONCLUSIONS: Childhood vitiligo has its own clinical features. The different types of vitiligo have different characteristics. There is immunological dysfunction in children with vitiligo. Dysfunction of humoral immunity may play a role in the progression of segmental vitiligo, while non-segmental vitiligo is more related to cellular immunity.

Double-stranded RNA induces melanocyte death via activation of Toll-like receptor 3.

As cutaneous pigment-producing cells, melanocytes can become targets of primary and secondary immune response as can be seen in diseases like vitiligo and Vogt-Koyanagi-Harada (VKH) syndrome. Viral infections have previously been implicated as a possible precipitating factor in the destruction of melanocytes in these disorders. During viral replication, double-stranded RNA (dsRNA) is produced as an intermediate metabolite, which induces antiviral and inflammatory responses through Toll-like receptor 3 (TLR3) in cells of innate immune system. The functional responses of melanocytes to dsRNA, however, remain unclear. Herein, we demonstrated that human melanocytes expressed TLR3 at a constitutive and inducible level. Stimulation with poly(I:C), a synthetic dsRNA analogue, triggered apoptosis of melanocytes. The apoptosis-inducing effect was shown by RNA interference to be largely dependent on TLR3, but occurred independently of NF-κB activation since treatment with specific NF-κB inhibitor Bay 11-7082 failed to prevent the process. In contrast, IFN-ß neutralizing Ab blocked the apoptosis-inducing effect of dsRNA, indicating the involvement of IFN-ß autocrine signalling. Furthermore, studies on the intracellular signal transduction pathways revealed that dsRNA induces the activation of p38, ERK1/2 and JNK1/2 in melanocytes. Using specific inhibitors, we demonstrated that activation of p38 and ERK1/2 controlled both IFN-ß secretion and IFN-ß mediated cell death. Taken together, these data suggest that viral dsRNA stimulates TLR3 in human melanocytes and triggers the cellular apoptosis through autocrine of IFN-ß.

Cutaneous infectious granuloma caused by Phenylobacterium in an adult with myelodysplastic syndrome: a first case report.

Painful granulomatous lesions appeared on the face of a 36-year-old man with myelodysplastic syndrome. Skin biopsy revealed chronic inflammatory granuloma. Bacterial cultures of the lesions and blood indicated the same unknown Gram-negative rod bacterium. The 16S ribosomal RNA sequence of the unknown bacterium yielded Phenylobacterium. Thus, we diagnosed cutaneous infectious granuloma caused by Phenylobacterium and myelodysplastic syndrome/refractory cytopenia with multi-lineage dysplasia. After treatment with combined antibacterials that were selected based on the tests for drug sensitivity, the lesions disappeared with only scars remaining and without any signs of relapse after 1 year. This is the first case report of cutaneous infectious granuloma caused by Phenylobacterium.

An unusual case of granulomatous lymphadenitis due to Prototheca zopfii var. portoricensis in an immunocompetent man in China.

BACKGROUND: Protothecosis is an uncommon human infection caused by Prototheca. Prototheca spp can be considered as saprophytes, and in spite of their frequency in the environment, they are of low virulence and may cause chronic infection with low-grade inflammation in humans. At present, only three species are recognized: Prototheca wickerhamii, Prototheca zopfii and Prototheca stagnora. Of these, the former two have been associated with human disease. This study was an investigation of the clinical and microbiological features of a case of granulomatous lymphadenitis due to P. zopfii var. portoricensis in an immunocompetent man in China. METHODS: We report the case of a 39-year-old male, who presented with swollen lymph nodes, from which the organism was isolated and identified by the RapidID Yeast Plus test (Remel, Santa Fe, NM, USA) and PCR molecular analysis. The pathogenicity of the isolate was confirmed in a mouse model and antifungal drug susceptibility testing was carried out. RESULTS: The pathogen was identified as Prototheca zopfii. The DNA sequence of the 18S SSU rDNA regions of the isolate strain were 100% (1205/1205) identical with Prototheca zopfii var. portoricensis. Antifungal susceptibility tests revealed that it was sensitive to amphotericin B, but resistant to 5-flucytosine, fluconazole, ketoconazole, and itraconazole. The patient responded to treatment with intravenous itraconazole and amphotericin B. CONCLUSIONS: Based on the patient's symptoms and microscopic evaluation, cultures, and molecular analyses of the isolate, granulomatous lymphadenitis due to P. zopfii var. portoricensis was diagnosed. P. zopfii var. portoricensis as a causative agent of human lymphadenitis in an immunocompetent case has not been reported, though a few cases of protothecosis have been reported in China. The real number of protothecosis cases may be greater than that reported in the literature. Thus, clinicians should be vigilant for any unknown cause of granulomatous lymphadenitis and should undertake an intensive histopathology, mycology examination, and even molecular analysis to rule out or confirm a potential Prototheca infection.

Tinea capitis in Southeastern China: a 16-year survey.

This survey was a retrospective of a 16-year (1993-2008) study on the incidence, clinical features, and etiological agents of tinea capitis mainly representing the Southeastern China. The diagnosis was confirmed by direct microscopic examination. Eight hundred and sixty-six patients with tinea capitis, 381 males (44%) and 485 females (56%), were enrolled in this study. Patients were between 20 days and 84 years old with an average of 10.5 years and the peak incidence was in the age group of 6-10 (48.5%). Five hundred and sixty-two patients (64.9%) were ectothrix and 303 patients (35.0%) were endothrix with only one patient was favus. The incidence of tinea capitis from 1993 was gradually increasing and reaching to its peak in 2001. Positive cultures of dermatophytes were obtained in 715 patients: Microsporum canis (62.4%) was predominant, followed by Trichophyton violaceum (19.0%), Trichophyton tousurans (9.8%). M. canis was the major pathogen for ectothrix infection, while T. violaceum and T. tousurans contributed to the most endothrix form. M. canis, T. violaceum, and T. rubrum were the major pathogens for kerion.

Reassessment of microarray expression data of porokeratosis by quantitative real-time polymerase chain reaction.

BACKGROUND: Porokeratosis (PK) is a heterogeneous group of keratinization disorders that exhibit similarities with psoriasis at both the clinical and molecular levels. METHODS: The transcript levels of keratin (KRT) 6A, 16, 17, S100A7, A8, A9, p53 and three candidate genes (i.e. SART3, SSH1 and ARPC3) were reassessed in pairwise lesional and uninvolved skin from nine patients with PK by real-time quantitative polymerase chain reaction (RTQ-PCR). RESULTS: The results of RTQ-PCR confirmed that KRT6A, 16, S100A7, A8 and A9 (p = 0.008) were mostly up-regulated in the lesional skin when compared with uninvolved skin. Different from the microarray data, there was no significant difference observed in KRT17 expression patterns between lesional and normal-appearing skin (p = 0.066). No statistical difference was observed in p53 and three candidate genes' expression patterns between lesional and uninvolved skin. CONCLUSIONS: In the present study, 9 of the 10 gene expression measured by RTQ-PCR in PK were statistically comparable to microarray data. KRT6A was identified as specific biomarker for porokeratotic keratinocytes, as it was the most significantly up-regulated gene in the nine patient samples.

Reversal of ultraviolet B-induced immunosuppression by inhibition of the extracellular signal-regulated mitogen-activated protein kinase.

OBJECTIVE: Topical treatment of the specific inhibitor PD98059 (PD) for extracellular signal-regulated kinase (ERK)1/2 combined with ultraviolet B (UVB) exposure in an in vivo study was proposed to confirm the effectiveness of ERK1/2 involved in UVB-induced immunosuppression that was reversed by PD. METHODS: Based on the mouse model of local UVB-induced immunosuppression [UVB exposure, followed by sensitization with dinitrofluorobenzene (DNFB) on the abdomen skin before challenge on the ear site], the PD was applied on the abdomen-irradiated area 1 h, immediately before and 6 h after UVB exposure, respectively. The baseline of ear thickness was measured and remeasured 24 h after the challenge of DNFB for evaluation of ear-swelling response. Histopathologically, the ear biopsies were taken for hematoxylin and eosin staining. RESULTS: Mice that received PD post-irradiation treatment showed a statistically significant contact hypersensitivity compared with the UVB-irradiated mice (P<0.05), and paralleled with the biopsy showing a thickened epidermis with lymphocyte infiltration. Thus, the PD had abrogated the UV-induced local suppression of contact hypersensitivity. CONCLUSION: The ERK1/2 mitogen-activated protein kinase (MAPK) pathway plays an important role in the local UVB-induced immunosuppression, and its specific inhibitor PD can arrest its function, resulting in protection against UVB-induced immunosuppression in the present in vivo study.

Cultured human melanocytes express functional toll-like receptors 2-4, 7 and 9.

BACKGROUND: Toll-like receptors (TLRs) recognize pathogen-associated molecular patterns (PAMPs), which have been evolutionarily conserved in microbes. Human melanocytes are not simply pigment-producing cells but also have the phagocytic capacity and can produce pro-inflammatory mediators. However, the mechanisms of recognition of microbes by melanocytes have not yet been fully established. OBJECTIVE: We investigated the TLRs 1-10 expression profile in human epidermal melanocytes and assessed their functions after triggering by their specific ligands. METHODS: TLRs mRNA expression was determined by RT-PCR, and the TLR protein expression was measured by flow cytometry and immunofluorescence assays. After stimulation with various TLR ligands, the production of inflammatory cytokine IL-8 and IL-6 was measured by ELISA and the mRNA for chemokine CCL2, CCL3 and CCL5 was analyzed by real-time PCR. Phosphorylation of IkappaBalpha in TLR ligands-triggered melanocytes was determined by Western blot and the nucleus translocation of NF-kappaBp65 was analyzed by immunofluorescence. RESULTS: Human melanocytes constitutively expressed TLRs 1-4, 6, 7 and 9 mRNA. Ample amounts of TLRs 2-4, 7 and 9 were confirmed at protein level. Stimulation of melanocytes with TLRs ligands resulted in the release of cytokines (IL-8 and IL-6) and the mRNA accumulation of chemokines (CCL2, CCL3 and CCL5). Triggering of TLRs in melanocytes resulted in the up-regulation of phosphorylated IkappaBalpha and in the nucleus translocation of NF-kappaBp65. CONCLUSION: Present study indicates human melanocytes express a panel of functional TLRs. The ligation of TLRs can turn these cells into active players of the skin innate immunity.

Primary cutaneous mucormycosis caused by Rhizomucor variabilis in an immunocompetent patient.

Primary cutaneous mucormycosis is an uncommon disease and occurs mainly in patients with immunocompromised disorders. We report a case of cutaneous mucormycosis in an immunocompetent man in whom no definite precipitating factors were noted. The isolate was identified as Rhizomucor variabilis according to the fungus morphology and DNA sequencing results. The lesion was successfully treated with oral Itraconazole, although the in vitro drug-susceptibility test showed resistance.

The susceptibility to vitiligo is associated with NF-E2-related factor2 (Nrf2) gene polymorphisms: a study on Chinese Han population.

Vitiligo is an acquired pigmentary disorder and its pathogenesis remains unclear. Oxidative stress is considered to be the initial pathogenic event in the melanocyte destruction. NF-E2-related factor2 (Nrf2) is a transcription factor regulating the expression of detoxifying and antioxidant genes. To investigate the association of the Nrf2 gene promoter polymorphisms with vitiligo in Chinese Han population, the genotypes of -686A/G, -684G/A and -650C/A and the genotyping of variable number of tandem repeat were detected. The data were analysed by the chi-square test and the risk was evaluated by calculating OR and 95% CI. There was statistically significant difference in genotypic and allelic frequencies of -650C/A between the two groups (P < 0.05). A(-650) allele was significantly associated with the risk for vitiligo (OR = 1.724, chi(2) = 18.096). Polymorphism of the Nrf2 gene promoter at -650C/A was associated with the development of vitiligo and A(-650) allele may be one of the risk factors.

Spontaneous regression of generalized eruptive histiocytosis: possible involvement of apoptosis?

Generalized eruptive histiocytosis (GEH) is a rare generalized non-X histiocytosis, first described in 1963 by Winklemann and Muller. Since then more than 20 patients with GEH, mainly adults but also a few children, have been reported. We report a case of GEH in a middle-aged woman with spontaneous regression, in which the ultrastructural findings of apoptosis were observed.

Inhibition of monocyte-derived dendritic cell differentiation and interleukin-12 production by complement iC3b via a mitogen-activated protein kinase signalling pathway.

We have previously demonstrated that iC3b is deposited at the dermal-epidermal junction of the skin following ultraviolet (UV) exposure and that it plays a role in UV-induced immunosuppression and antigenic tolerance. In vitro, iC3b differentially regulates monocyte production of interleukin-10 (IL-10) and IL-12. Additionally, iC3b arrests monocytic cell differentiation into CD1c-expressing dendritic cell (DC) precursors. The present study addresses mitogen-activated protein kinase (MAPK) signalling following the cross-linking of CR3 by its ligand iC3b with regard to monocyte differentiation and cytokine regulation. Sheep erythrocytes were coated with IgM alone (EA) or iC3b (EAiC3b) to allow for CR3 cross-linking onto monocytes. EAiC3b increased the phosphorylation (p) of extracellular signal-regulated kinase (ERK) MAPK in fresh human monocyte, particularly in monocyte-derived DC (MDDC) that were differentiated by means of GM-CSF (1000 U/ml) and IL-4 (200 U/ml) for 2 days before iC3b exposure for an additional 24 h (P=0.034, n=3). CD1a expression, induced by GM-CSF and IL-4, was inhibited by iC3b (EAiC3b vs. EA, P=0.012, n=4). Conversely, the inhibition of ERK by the specific inhibitor (PD98059), but not the p-38 inhibitor SB203580, restored CD1a expression (P=0.011, n=4) in iC3b-stimulated MDDC. Concordantly, the inhibition of ERK during iC3b exposure fully reversed the inhibition of IL-12p70 induction in MDDC by 95% (P<0.01, n=4) and decreased IL-10 production. Taken together, our data demonstrate that iC3b interferes with MDDC differentiation and IL-12 and IL-10 production is mediated via an ERK MAPK-dependent mechanism. Thus, ERK MAPK inhibition may represent a therapeutic strategy for preventing monocytic precursor diversion away from DC differentiation when monocytes enter injured tissues in which iC3b is generated, such as UV-exposed skin.

Inhibition of monocytic interleukin-12 production by Candida albicans via selective activation of ERK mitogen-activated protein kinase.

Our previous data demonstrated that live Candida albicans inhibits interleukin-12 (IL-12) production by human monocytes. Here we explored whether C. albicans inhibits IL-12 via a released factor and whether the inhibition is mediated via mitogen-activated protein kinase (MAPK) regulation. Supernatant fluids were obtained from cultured C. albicans (SC5314) as well as cultured Saccharomyces cerevisiae after 20 h of incubation. At 2 h postincubation of monocytes with heat-killed C. albicans (HKCA) (2:1) to stimulate IL-12, concentrated fungal supernatant fluids were added and incubated for an additional 20 h. The present data show that, unlike supernatant fluids obtained from S. cerevisiae, the C. albicans supernatant fluids significantly suppressed IL-12 production induced by HKCA. This suggested that the inhibition is Candida specific. A preliminary biochemical analysis revealed that this secretory IL-12 inhibitory factor is glycoprotein in nature. The inhibitory activity had no effect on the phagocytosis of yeasts. Supernatant fluids from C. albicans markedly induced the phosphorylation of ERK44/42 MAPK, but not p38 and SAPK, 1 min after they were added to monocytes. To test if the induction of ERK44/42 MAPK was central to the IL-12 inhibition, we used gamma interferon (IFN-gamma) (1 ng/ml) plus lipopolysaccharide (LPS) (100 ng/ml) to stimulate IL-12 production by monocytes. The inhibition of ERK MAPK by the specific inhibitor PD 98059 significantly reduced phospho-ERK44/42 MAPK levels induced by C. albicans supernatant fluids in the IFN-gamma-plus-LPS-driven monocytes. Concomitantly, PD 98059 reversed the IL-12 inhibitory activity of the C. albicans supernatant (P < 0.01). These data indicate that C. albicans can inhibit IL-12 production by secreting an ERK44/42 MAPK-stimulating factor and thus can attenuate effective immune responses.