Objective To explore the effect of curcumin on the activity and migration of as well as c-kit mRNA expression in melanocytes.Methods Human epidermal melanocytes were isolated from the prepuce in adolescents and subjected to primary culture.To estimate the effect of curcumin on the proliferative activity of melanocytes, some melanocytes were randomly divided into several groups to be cultured in the MelM-2 medium with or without the presence of 5, 10, 20 or 30 μmol/L curcumin, the MelM-2 medium containing curcumin of 5-30 μmol/L served as the drug control groups, and the MelM-2 medium without curcumin served as the blank control group.After 24 and 48 hours of culture, MTS assay was performed to evaluate the proliferative activity of melanocytes.Some cultured melanocytes were randomly divided into 4 groups to be cultured in the MelM-2 medium with 0, 5, 10 and 20 μmol/L curcumin respectively for 48hours.Then, wound scratch assay was conducted to estimate the migratory activity of melanocytes, and real-time fluorescence-based quantitative PCR to quantify the mRNA expression of c-kit in melanocytes.Statistical analysis was carried out by factorial design analysis of variance (ANOVA), one-way ANOVA and least significant difference (LSD)-t test.Results The proliferative activity of melanocytes was significantly decreased at 24 and 48 hours in the 30-μmol/L curcumin group compared with the negative control group (0.783 ± 0.053 vs.1.000 ± 0.018 at 24 hours, 0.637 ± 0.015 vs.0.993 ± 0.064 at 48 hours, both P ＜ 0.05), while no significant differences were observed between the other curcumin groups and the negative control group (all P ＞ 0.05).The 48-hour treatment with curcumin could significantly inhibit the migration of melanocytes in the 5-, 10-and 20-μmol/L curcumin groups compared with the control group (all P ＜ 0.05).The mRNA expression level of c-kit was also significantly reduced at 48 hours in the 5-, 10-and 20-μmol/L curcumin groups compared with the control group (1.799 ± 0.372, 1.539 ± 0.224 and 1.026 ± 0.038 vs.3.371 ± 0.352, all P ＜0.05).Conclusion Curcumin at low concentrations (≤ 20 μmol/L) has no obvious cytotoxicity against melanocytes, but can inhibit the migration of and c-kit mRNA expression in melanocytes, while curcumin at 30 μmol/L can promote the apoptosis of melanocytes.

Objective To evaluate the effect of tacalcitol on the proliferation,adhesion,migration and c-kit mRNA expression of cultured human epidermal melanocytes.Methods Cultured epidermal melanocytes from the prepuce of adolescent males were treated with various concentrations of tacalcitol.Then,cell proliferation was evaluated by tetrazolium salt (XTT) assay after 24,48 and 72 hours of treatment,adhesive activity by using fibronectin-coated culture plates after 72 hours,migratory activity by Transwell assay using a microporous membrane after 24 hours,and the c-kit mRNA expression was semiquantitatively analyzed by reverse transcription PCR after 72 hours of treatment.Statistical analysis was done by repeated-measure analysis of variance and completely random design analysis of variance.Results As repeated-measure analysis of variance showed,tacalcitol of 10-10,10-9,10-8,10-7 and 10-6 mol/L significantly promoted the proliferation of melanocytes (F =9.47,P ＜ 0.01),with significant differences in the promoting effect among various durations of treatment with different concentrations of tacalcitol (F =14.44,P ＜ 0.01),and with significant interaction effect between drug concentration and treatment duration (F =2.47,P ＜ 0.01).The highest proliferation level was observed in melanocytes treated with tacalcitol of 10-s mol/Lfor 72 hours.There was a significant increase in the adhesion rate of human epidermal melanocytes to fibronectin after treatment with tacalcitol of 10-8-10-7 mol/L for 72 hours (both P ＜ 0.01),number of melanocytes migrating through micropore membranes per high-power field (× 200) after treatment with tacalcitol of 10-9-10-8 mol/L for 24 hours (both P ＜ 0.01),and in the c-kit mRNA expression in melanocytes treated with tacalcitol of 10-9-10-7mol/L for 72 hours (all P ＜ 0.01).Conclusion Tacalcitol can promote melanocytes to proliferate,migrate,express c-kit mRNA,and adhere to fibronectin.

A total of 107 vitiligo patients were randomly divided into 3 groups.Group A received an intralesional injection of Bacillus Calmette-Guérin-polysaccharide nucleic acid (BCG-PSN) (n =34),group B Compound Glycyrrhizin Tablets (n =36) and group C both (n =37).Before treatment and 3 months after treatment,cellular immune function was detected for each group.Paired comparisons of 3 groups before and after treatment showed that CD4 +,CD4 +/CD8 + ratio increased (all P ＜ 0.05) and CD8 + decreased (P ＜0.05).After treatment,as compared with groups A and B,CD4 + increases (both P ＜ 0.05) and CD8 + decreased in group C (P ＜0.05).Group C had an efficiency rate of 91.9％ and it was higher than the other two groups (both P ＜ 0.05).An intralesional injection of BCG-PSN plus Compound Glycyrrhizin Tablets could improve immune function and treat vitiligo patients efficiently.

Objective To study the regulatory effect of ethanol extract of glossy privet fruit and its monomer tyrosol on the adhesion and migration of human epidermal melanocytes.Methods Epidermal melanocytes were isolated from human foreskin,and subjected to a primary culture.Mter 3-5 passages,the melanocytes were treated with various concentrations of ethanol extract of glossy privet fruit (0.0375-0.6 mg/ml)and tyrosol (0.125-2 mmol/L) for 24-72 hours.The XTT colorimetric assay was carried out to evaluate the proliferation of melanocytes,fibronectin (FN)-coated culture plates were used to evaluate the adhesion activity of melanocytes,and Transwell assay was conducted to assess the migration activity of melanocytes.Confocal laser microscopy was utilized to observe the structure and distribution of actin cytoskeleton in melanocytes,and cellular fluorescence intensity was determined by a semi-quantitative analysis.Statistical analysis was carried out by using unpaired t test.Results The adhesion activity of melanocytes to FN was significantly enhanced by the ethanol extract of 0.0375-0.6 mg/ml from glossy privet fruit (P ＜ 0.05 or 0.01),and by tyrosol of 0.5-2 mmol/L (P ＜ 0.05 or 0.01).As XTT assay showed,neither the ethanol extract of 0.15 mg/ml nor tyrosol of 2 mmol/L had cytotoxicity or promotive effect on cell proliferation.Hence,0.15 mg/ml and 2 mmol/L were determined as the working concentrations of ethanol extract and tyrosol respectively.The number of cells migrating through micropore membranes per high-power field (× 200) was 43.7 and 51.0 in melanocytes treated with the ethanol extract of 0.15 mg/ml and tyrosol of 2 mmol/L,respectively,significantly higher than that in untreated melanocytes (20.3,both P ＜ 0.01).Compared with untreated melanocytes,those treated with the ethanol extract of 0.15 mg/ml and those with tyrosol of 2 mmol/L showed higher intracellular fluorescence intensity (P ＜ 0.01) and more stress fiber bundles which congregated inside the cell membrane and around the nuclei.Conclusions The ethanol extract of glossy privet fruit and its monomer tyrosol can promote the adhesion and migration of human melanocytes in vitro,likely by promoting the congregation of actin cytoskeleton in melanocytes.

Objective:To explore the value of radiomics model based on intratumoral and peritumoral early dynamic contrast-enhanced (DCE) MRI for identifying benign and malignant in breast imaging reporting and data system (BI-RADS) 4 tumors.Methods:A total of 191 patients diagnosed with BI-RADS 4 breast tumors by breast MRI examination with clear pathological diagnosis from January 2016 to December 2020 in the First Affiliated Hospital of Bengbu Medical College were analyzed retrospectively, including 77 benign and 114 malignant cases, aged 23-68 (46±10) years. The one-slice image with the largest area of the lesion of the second stage DCE-MRI images was selected to outline the region of interest, and automatically conformal extrapolated by 5 mm to extract the intra-tumoral and peritumoral radiomics features. The included cases were randomly divided into training and testing cohorts in the ratio of 8∶2. The statistical and machine learning methods were used for feature dimensionality reduction and selection of optimal radiomics features, and logistic regression was used as the classifier to establish the intratumoral, peritumoral, and intratumoral combined with peritumoral radiomics models. The independent risk factors that could predict the benignity and malignancy of breast tumors were retained as clinical-radiological characteristics by univariate and multivariate logistic regression to establish a clinical-radiological model. Finally, the intratumoral and peritumoral radiomics features were combined with clinical-radiological features to develop a combined model of the three. The receiver operating curve was used to analyze the predictive performance of each model and calculate the area under the curve (AUC),the AUC was compared by DeLong test. The stability of the three-component combined diagnostic model was tested by 10-fold cross-validation, and the model was visualized by plotting nomogram and calibration curves.Results:In the training cohort, the AUC of the three-component combined model for identifying benign and malignant BI-RADS 4 breast tumors was significantly higher than that of the intratumoral radiomics model ( Z=3.38, P<0.001), the peritumoral radiomics model ( Z=4.01, P<0.001), the intratumoral combined with peritumoral radiomics model ( Z=3.11, P=0.002), and the clinical-radiological model ( Z=3.24, P=0.001). And the AUC, sensitivity, specificity, accuracy, and F1-score of the three-component combined model were 0.932, 91.2%, 86.9%, 87.0% and 0.89, respectively. In the testing cohort, the three-component combined model also had the highest AUC value (0.875), and diagnostic sensitivity, specificity, accuracy and malignancy F1-score were 95.7%, 62.5%, 76.9%, and 0.89, respectively. The AUC calculated by 10-fold cross-validation was 0.90 (0.85-0.92), and the predicted curve of the three-component combined model in the calibration curve was in good agreement with the ideal curve. Conclusion:The three-component combined diagnostic model based on the intratumoral and peritumoral radiomics features and clinical-radiological features of early DCE-MRI has good performance and stability for identifying the benign and malignant in BI-RADS 4 breast tumors, and it can provide guidance for clinical decision non-invasively.