Objective To investigate the morphology and quantity of melanin and blood vessels in chloasma lesions by using reflectance confocal microscopy(RCM)and dermoscopy, and to explore the relationship of melanin and blood vessels with lesion color. Methods RCM was used to evaluate the quantity and distribution of melanin in lesions of 135 patients with chloasma, and to observe the morphology of melanin and dendritic melanocytes in 54 of the 135 patients, and dermoscopy was performed to assess the quantity and morphology of blood vessels in lesions of the 54 patients. Statistical analysis was carried out by Spearman correlation analysis to assess the relationship of melanin and blood vessels with lesion color. Results Increased quantity of melanin was observed in both the epidermal layer and dermo?epidermal junction in all the 135 patients, and in only the superficial dermis in 36(27%)patients. No patients showed increased quantity of melanin in only the dermis without epidermal involvement. Among the 54 patients, there was a positive correlation between the quantity score and morphology score of melanin(r=0.73, P<0.001), between the quantity score and morphology score of blood vessels(r=0.87, P<0.001), as well as between the total melanin score and total blood vessel score(r=0.554, P<0.001). Lesion color was positively correlated with the quantity and morphology of melanin(r = 0.51, 0.39, respectively, both P < 0.05)and blood vessels(r = 0.46, 0.44, respectively, both P<0.05). Conclusions RCM and dermoscopy can be used to evaluate melanin and blood vessels in chloasma lesions. There is a positive correlation between the quantity and morphology of melanin and between those of blood vessels in chloasma lesions, and lesion color is positively correlated with the quantity and morphology of melanin and blood vessels.

Objective To explore the effects of alcoholic extracts of traditional Chinese medicines on the post-translational processing and trafficking of tyrosinase.Methods Human YUGEN8 amelanotic melanoma cells were grown in vitro;the cells were incubated with one of the seven traditional Chinese medicines,including Rhizoma Chuanxiong and psoralen.Protein analysis with Western blot,enzymolysis with endoglycosidase H (Endo H),and subcellular localization with laser confocal microscopy were per- formed.The expression,maturity and export from endoplasmic reticulum (ER) of tyrosinase in the treated cells were compared with those in the untreated controls.Results Compared with controls,an approximate- ly 80-kDa,Endo H-resistant tyrosinase doublet,which represented mature glycoform of tyrosinase,was in- creased in melanocytes treated with Semen Cuscutae,and in those treated with Semen Persicae.Within those cells,tyrosinase was distributed outside ER resident protein calnexin.Conclusion Both Semen Cus- cutae and Semen Persicae could induce tyrosinase maturation,stability and export from ER to distal site.

BACKGROUNDThe purpose of this study was to evaluate the efficacy of topical melagenine for repigmentation in child patients with vitiligo on the scalp.

METHODSTwenty-two child patients with vitiligo on the scalp were treated with 1.2 mg/ml aqueous melagenine in combination with 20 minutes of infrared exposure twice daily.

RESULTSIn 4 patients (18.2%), melagenine treatment in combination with infrared exposure led to complete recovery; in 6 patients (27.3%), treatment was shown to be effective; in 8 patients (36.3%), treatment led to improvements in patient condition; and only 4 patients (18.2%) showed no response after 1 - 2 treatment sessions. The general effective rate of melagenine-infrared combination treatment was 45.5% for the children with vitiligo on the scalp, and treatment was accompanied by minimal side effects.

CONCLUSIONMelagenine may be efficacious and a safe treatment option for childhood vitiligo affecting the scalp.

Administration, Topical ; Adolescent ; Child ; Child, Preschool ; Female ; Humans ; Infrared Rays ; therapeutic use ; Lipoproteins ; administration & dosage ; Male ; Scalp Dermatoses ; drug therapy ; Vitiligo ; drug therapy

OBJECTIVETo explore the correlation of the gene polymorphisms of Toll-like receptor 2 ( TLR2) and TLR4 with the susceptibility and recurrence of condyloma acuminatum (CA).

METHODSUsing Snapshot, we detected the gene polymorphisms of TLR2 597(T/C), 1350(T/C), 15607(A/G), and 2258(G/A) and TLR4 896(A/G) and 1196(C/T) in the peripheral blood of 140 CA patients and 105 HPV-negative controls. We made comparisons between the CA patients and controls as well as between the cases of recurrent CA and those of non-recurrence at 6 months after treatment.

RESULTSThere were 72, 48, and 20 cases of genotype TT, TC, and CC of TLR2 597 (T/C), respectively, in the CA patients, as compared with 71, 31, and 3 cases in the controls. The gene frequency of mutant C was 31. 43% in the patients, significantly higher than 17.62% in the controls (χ2 = 12.04, P < 0.01), and it was 38.68% in the recurrent cases, remarkably higher than 27.01% in the non-recurrent cases (χ2 = 4.16, P < 0.05). There were 74, 49, and 17 cases of genotype TT, TC, and CC of TLR2 1350( T/C), respectively, in the CA patients, as compared with 73, 29, and 3 cases in the controls. The gene frequency of mutant C was 29. 64% in the patients, significantly higher than 16. 67% in the controls (χ2 =11.05, P < 0.01), and it was 36.79% in the recurrent cases, markedly higher than 25. 29% in the non-recurrent cases (χ2 = 4.18, P < 0.05). There were 44, 66, and 30 cases of genotype AA, AG, and GG of TLR2 15607(A/G), respectively, in the CA patients, as compared with 26, 58, and 21 cases in the controls. There was no significant difference in the gene frequencies of mutant G between the two groups (χ2 = 0.33, P > 0.05). No mutant genes of TLR2 2508 (G/A) or TLR4 896(A/G) and 1196(C/ T) were detected in either the CA patients or the controls. Linkage disequilibrium analysis showed a tight linkage between TLR2 597 (T/C) and 1350(T/C) (D' = 1, r2 = 0.93).

CONCLUSIONTLR2 597(T/C) is tightly linked to 1350(T/C), which is correlated with both the susceptibility and the recurrence of condyloma acuminatum.

Aged ; Case-Control Studies ; Condylomata Acuminata ; genetics ; Gene Frequency ; Genetic Linkage ; Genetic Predisposition to Disease ; Genotype ; Humans ; Polymorphism, Genetic ; Recurrence ; Toll-Like Receptor 2 ; genetics ; Toll-Like Receptor 4 ; genetics

Objective:To investigate the efficacy of systemic glucocorticoid treatment and its related factors in progressive vitiligo patients with vitiligo disease activity (VIDA) scores ≥ 2 points.Methods:A total of 272 progressive vitiligo patients with VIDA scores ≥ 2 points and skin lesion area < 1% of body surface area, who received no systemic glucocorticoid treatment, were collected from Department of Dermatology, the Third People′s Hospital of Hangzhou from June 2018 to June 2019. The area and type of skin lesions, VIDA scores, predisposing factors and special clinical markers (trichrome vitiligo, confetti-like depigmentation, Koebner phenomenon and inflammatory vitiligo) were analyzed. These patients were randomly divided into 3 groups by a random number table: topical glucocorticoid group (62 cases) , oral prednisone + topical glucocorticoid group (76 cases) and compound betamethasone injection + topical glucocorticoid group (134 cases) , and the latter two groups were also called as the systemic and topical glucocorticoid group. The patients in the topical glucocorticoid group were treated with halometasone cream or 0.05% clobetasol propionate cream once a day; during the oral prednisone treatment, the dose was adjusted once every 7 days, and gradually reduced from 30 mg/d to 20, 15, 10 and 5 mg/d, and the treatment lasted 35 days; during the treatment with compound betamethasone injection, intramuscular injection was performed once every 20 days at a dose of 1 ml for 2 sessions. The stable disease rate (defined as the proportion of patients experiencing no progression during the study among the analyzed patients) was calculated in these groups after 3 months of treatment, and changes in vitiligo types were evaluated after 1 year of follow-up. Statistical analysis was carried out by using Kruskal-Wallis H test, χ2 test and Fisher′s exact test. Results:After 3-month treatment, there was a significant difference in the expansion rate of skin lesion area among the 3 groups ( H = 12.468, P < 0.001) , and the expansion rate of skin lesion area was significantly lower in the oral prednisone + topical glucocorticoid group and compound betamethasone injection + topical glucocorticoid group than in the topical glucocorticoid group ( P < 0.001, = 0.005, respectively, α = 0.016 7) ; among the patients with slowly progressive vitiligo (VIDA scores = 2 or 3 points) , the stable disease rate was significantly higher in the systemic and topical glucocorticoid group than in the topical glucocorticoid group ( χ2 = 23.973, 11.877, respectively, both P < 0.001) ; the stable disease rate also significantly differed among the patients with different VIDA scores (VIDA scores = 2, 3 or 4 points) in the systemic and topical glucocorticoid group ( χ2 = 17.122, P < 0.001) . After 3-month treatment, the patients with predisposing factors or special clinical markers showed significantly decreased stable disease rate (47.3% [35/74], 41.2% [47/114], respectively) compared with those without predisposing factors or special clinical markers (70.6% [96/136], 87.5% [84/96]; χ2 = 11.098, 47.548, respectively, both P < 0.001) . After 1 year of follow-up, the proportion of patients with localized vitiligo converted into non-localized vitiligo was significantly higher in the topical glucocorticoid group (41.9%, 26/62) than in the systemic and topical glucocorticoid group (21.9%, 46/210; χ2 = 10.328, P = 0.006) , and higher in the group with predisposing factors or special clinical markers than in that without predisposing factors or special clinical markers respectively (both P < 0.01) . Conclusions:Early systemic glucocorticoid treatment should be performed in the progressive vitiligo patients with high VIDA scores, predisposing factors and special clinical markers.

Objective To investigate the expression of VIT-1 gene in melanocytes of patients with vitiligo, and to analyze the difference of its sequence. Methods The skin from the foreskins of healthy men by circumcision and from the non-lesional area on the buttocks of 5 patients were digested by dispase, then the epidermis and dermis were separated, and the melanocytes were isolated. Then we cultured the melanocytes from the controls in TICVA medium and those from the patients in TICVA medium supplemented with basic fibroblast growth factor (bFGF) and endothelin-1 ( ET-1). The expression of VIT-1 gene was measured by RT-PCR, the full-length cDNA of VIT-1 ORF was cloned and sequenced, and sequence difference was analyzed by CLUSTAL W ( 1.83 ) software. Results The expression levels of VIT-1 gene were significantly lower in melanocytes from the patients than in those from the controls. An 81 bp-intron was found in the VIT-1 ORF. VIT-1 was a fragment of FBXO11, located at its 3' end. Conclusion VIT-1 gene is not a new gene, but a fragment of FBXO11, and a member of F-box protein family.

Objective To determine the effects of 1320 nm non-ablative laser on the proliferation of human dermal fibroblasts,and the secretion of basic fibroblast growth factor(bFGF)and transforming growth factor-?1(TGF-?1)in vitro.Methods Human dermal fibroblasts were cultured,and irradiated three times by 1320 nm laser at a dose of 15,20 and 24 J/cm~2,respectively.The levels of bFGF and TGF-?1 were examined by ELISA at 0,24,48 and 72h after the irradiation.The number of fibroblasts before and after irradiation were determined.Results The number of fibroblasts and the secretion of bFGF both in- creased after the irradiation at the doses of 20 J/cm~2 and 24 J/cm~2(P

OBJECTIVETo explore the inhibition effect of RNA interference on the ICP4 expression and DNA replication of herpes simplex virus type 2 (HSV2).

METHODSFour pairs of siRNA targeted to HSV2 ICP4 gene and negative control siRNA were synthetized by chemical method, named as siRNA-1, siRNA-2, siRNA-3, siRNA-4 and siRNA-N respecticely. HSV2 HG52 was used to attack Vero cell after transfection overnight. Vero cell and supernatant were collected at 1d, 2d, 3d, 4d and 5d after virus attacking. Flurogenic quantitative reverse transcription polymerase chain reaction (FQ-RT-PCR)was used to detect the expression of HSV2 ICP4 mRNA, flurogenic quantitative polymerase chain reaction(FG-PCR) was used to detect the expression of HSV2 DNA and Western-Blot was used to detect the expression of HSV2 ICP4 protein.

RESULTSAll the four pairs of siRNA could significantly inhibit the expression of HSV2 ICP4 mRNA and protein, especially siRNA-2. The above siRNAs could significantly decrease HSV2 DNA copy number,too.

CONCLUSIONsiRNAs targeted to HSV2 ICP4 gene could significantly inhibit expression of HSV2 ICP4 mRNA and protein, and decrease HSV2 DNA copy number, suggesting that siRNA can inhibit HSV2 DNA replication through silencing ICP4 gene.

Gene Expression Regulation, Viral ; drug effects ; genetics ; Gene Silencing ; drug effects ; physiology ; Herpesvirus 2, Human ; drug effects ; genetics ; RNA Interference ; drug effects ; immunology ; RNA, Small Interfering ; pharmacology ; RNA, Viral ; drug effects ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo explore the Mechanism of nonablative skin rejuvenation.

METHODThe Kunming mice be used as subjects and divided into three groups (A, B, C). A, B, C groups were irradiated with 1 320 nm cooltouch laser (20 J/cm2) in the skin of left back; B and C groups were irradiated two and three times respectively; the skin of right back of A, B, C groups was adopted as control. The expression of bFGF and TGF-beta1 in the mouse skin was examined by the immunohistochemistry . The fibroblasts were isolated from the foreskin and cultured. One group is a control and other three ones are low, intermediate and high energetic groups respectively. The fibroblasts were irradiated by laser with 15 J/cm2 ,20 J/cm2 and 24 J/cm2 energy for three times. We examined the levels of bFGF and TGF-beta1 by ELISA in 0, 24, 48 and 72 hours.

RESULTSAccording to this research on immunohistochemistry result, there are significant differences in the expression of bFGF and TGF-beta1 between the group irradiated by three times and others (P < 0.01). The number of fibroblasts get increased after being irradiated by laser. The ELISA result indicates that the secretion of bFGF increased in the group of intermediate and high energetic level after laser irradiating and may reach the peak at 24 hours (P < 0.01). The amount of TGF-beta1 secretion, however, seems to get decreased in each group at all energetic levels, and at 24 hours it can reach the top level as well.

CONCLUSIONThe direct influence of laser on the fibroblasts is to promote secretion of bFGF and to inhibit secretion of TGF-beta1, while its influence on the tissue is to promote the secretions of the both. Nonablative skin rejuvenation not only can induce fibroblasts to secrete more bFGF but also induce the blood vessels to release cytokines which stimulate endothelial cell to express more of bFGF and TGF-beta1. Furthermore, fibroblastic proliferation can accelerate by laser's irradiating.

Animals ; Cells, Cultured ; Cosmetic Techniques ; Female ; Fibroblast Growth Factor 2 ; metabolism ; Fibroblasts ; cytology ; radiation effects ; secretion ; Laser Therapy ; Mice ; Mice, Inbred Strains ; Rejuvenation ; Skin ; cytology ; radiation effects ; Transforming Growth Factor beta1 ; metabolism

BACKGROUNDNitic oxide (NO) has been implicated in the pathogenesis of various inflammatory diseases, including sunburn and pigmentation induced by ultraviolet irradiation. Epigallocatechin-3-gallate (EGCG) is the major effective component in green tea and can protect skin from ultraviolet-induced damage. The purpose of this study was to investigate the protective mechanisms of EGCG on inducible nitric oxide synthase (iNOS) expression and NO generation by ultraviolet B (UVB) irradiation in HaCaT cells.

METHODSHaCaT cells were irradiated with UVB 30 mJ/cm 2 and pretreated with EGCG at varying concentrations. The iNOS mRNA was detected by reverse transcriptase polymerase chain reaction (RT-PCR) and NO production was quantified by spectrophotometric method. The expression of NF-kappaB P65 was measured by immunofluorescence cytochemistry staining.

RESULTSThe expression of iNOS mRNA and generation of NO in HaCaT cells were increased by UVB irradiation. EGCG down regulated the UVB-induced iNOS mRNA synthesis and NO generation in a dose dependent manner. The UVB-induced ctivation and translocation of NF-kappaB were also down regulated by EGCG treatment in HaCaT cells (P < 0.01).

CONCLUSIONSGreen tea derived-EGCG can inhibit and down regulate the UVB-induced activation and translocation of NF-kappaB, expression of iNOS mRNA and generation of NO respectively, indicating EGCG may play a protective role from UVB-induced skin damage.

Catechin ; analogs & derivatives ; pharmacology ; Cells, Cultured ; Gene Expression Regulation, Enzymologic ; drug effects ; Humans ; Keratinocytes ; metabolism ; radiation effects ; Nitric Oxide ; biosynthesis ; Nitric Oxide Synthase Type II ; genetics ; Protein Transport ; drug effects ; RNA, Messenger ; analysis ; Tea ; Transcription Factor RelA ; metabolism ; Ultraviolet Rays ; adverse effects