Objective To evaluate the effects of metformin on transforming growth factor-betal (TGF-β 1)-induced epithelial-mesenchymal transition (EMT) in and invasion of the human melanoma cell line 1205Lu.Methods In vitro cultured 1205Lu cells were divided into 3 groups to be treated with serumfree culture medium (blank control group),serum-free culture medium containing 5 ng/ml TGF-β 1 (TGF-β 1 group) and serum-free culture medium containing 5 ng/ml TGF-β 1 and 1 mmol/L metformin (TGF-β1 + metformin group),respectively.After 48-hour treatment,morphological changes of 1205Lu cells in the above groups were observed by using a microscope,and photos were taken at the same time.Transwell invasion assay was performed to estimate cellular invasive activity,Western blot analysis and real-time fluorescence-based quantitative RT-PCR were conducted to determine the mRNA and protein expression of N-cadherin and matrix metalloproteinase-9 (MMP-9),respectively.Statistical analysis was carried out by one-way analysis of variance (ANOVA) and least significant difference (LSD) test.Results Compared with the blank control group,the stimulation of TGF-β 1 could induce the epithelial-mesenchymal transition (EMT)-like morphological changes in 1205Lu cells,while TGF-β 1 combined with metformin could reverse the EMT-like morpho-logical changes.The number of 1205Lu cells crossing the transwell Matrigel per high-power field (× 200) was significantly higher in the TGF-β1 group (412.2 ± 13.427) than in the blank control group (194.1 ± 8.295) and TGF-β1 + metformin group (175.3 ± 8.693).Compared with the blank control group and TGF-β1 + metformin group,the TGF-β1 group also showed significantly increased mRNA and protein expression of N-cadherin (mRNA:6.678 ± 0.043 vs.1.000 ± 0.000,1.035 ± 0.015;protein:1.963 ± 0.016 vs.0.603 ± 0.029,0.207 ± 0.009) and MMP-9 (mRNA:5.351 ± 0.604 vs.1.000 ± 0.000,0.930 ±0.130;protein:1.243 ± 0.027 vs.0.575 ± 0.009,0.629 ± 0.008).Conclusion Metformin can obviously inhibit TGF-β1-induced EMT-like morphological changes in,the capacity to penetrate Matrigel-coated transwell chambers of and the mRNA and protein expression of N-cadherin and MMP-9 in 1205Lu cells.

Objective To evaluate the value of modified calcofluor white fluorescent staining in the histopathological diagnosis of subcutaneous mycosis,in order to provide a new method for histopathological diagnosis of subcutaneous mycosis.Methods A total of 21 lesional skin tissues were collected from patients with subcutaneous mycosis in the Affiliated Hospital of Chengde Medical University between 1987 and 2017,and embedded in paraffin.Then,each paraffin-embedded tissue section was cut into 4 4-μm-thick serial sections,and subjected to modified calcofluor white fluorescent staining,hematoxylin and eosin (HE) staining,periodic acid Schiff (PAS) staining and Gomori methenamine silver nitrate (GMS) staining respectively.Positive rates and staining outcomes were compared among the above staining methods.Statistical analysis was carried out with SPSS 19.0 software by using chi-square test for comparing the positive rates among the above 4 staining methods.Results Of 21 patients with fungal infections,14 (66.67％) were positive for modified calcofluor white fluorescent staining,5 (23.80％) for HE staining,6 (28.57％) for PAS staining,and 11 (52.38％) for GMS staining.The positive rate by modified calcofluor white fluorescent staining was significantly higher than that by HE staining and PAS staining (x2 =6.718,5.200,respectively,both P ＜ 0.05),while no significant difference was observed between the modified calcofluor white fluorescent staining and GMS staining (x2 =0.693,P =0.530).Conclusion The modified calcofluor white fluorescent staining is an accurate method for detecting fungi,and has a certain application value in the histopathological diagnosis of subcutaneous mycosis.

Tinea of vellus hair is caused by dermatophyte infection of vellus hairs, and commonly affects children. It usually occurs on the face, and clinically manifests as annular or semi-annular erythema gradually spreading to the surrounding area, with central clearing and a slightly elevating border covered with papules and papulovesicles. Intense inflammation, which may manifest as pustules, erosions, exudation, scales and crusts, can be observed in patients with severe tinea of vellus hair. Direct microscopy of fungi showed abundant hyphae and/or spores on vellus hairs. Topical antifungal therapy is usually ineffective, and systemic antifungal therapy should be considered. In order to reduce the high rate of missed diagnosis and misdiagnosis, and to improve clinicians′ understanding of this disease, this review summarizes the incidence, clinical manifestations, diagnosis and treatment of tinea of vellus hair.