Objective To explore the inhibitory action of antisense c-jun oligodeoxynucleotides(ODN) on matrix metalloproteinase(MMP) expression of fibroblasts induced by ultraviolet B (UVB). Methods The c-jun and c-fos protein expression induced by UVB were measured by Western blot. The mRNA expression of c-jun was determined by reverse transcriptase polymerase chain reaction (RT-PCR) after transfection with c-jun antisense ODN. The pro-MMP-1 and MMP-3 synthesis of fibroblasts was measured by ELISA after treatment with UVB and antisense c-jun ODN. Results The UVB-induced c-jun protein expression of fibroblasts increased to 1.8, 2.6, 3.3 times compared with that of non-irradiated controls,while there was no significant change in c-fos protein expression. The pro-MMP-1 and MMP-3 synthesis induced by UVB irradiation increased obviously. After transfection with different concentrations of c-jun antisense ODN, the UVB-induced c-jun mRNA expression could be significantly inhibited(P

Objective To investigate the protective effect of aloin on inducible nitric oxide synthase (iNOS) and nuclear factor kappa B (NF-kB) synthesis of HaCat cells induced by ultraviolet B (UVB) irradiation. Methods The proliferation of HaCat cells was measured by MTT method. iNOS mRNA was detected by reverse transcriptase polymerase chain reaction (RT-PCR). NO production was assessed by spectrophotometric method, and expression of NF-kB P65 was measured by immunofluorescent staining. Results After irradiation with 30 mJ/cm~2 of UVB, proliferation of HaCat cells was decreased, and NO generation and iNOS mRNA synthesis in HaCat cells were increased. UVB irradiation could also activate the expression of NF-kB P65 and promote its translocation into nucleus. NO generation and iNOS mRNA synthesis were markedly down-regulated in a dose-dependent manner by pre-treatment with different concentrations of aloin. The activation of NF-kB P65 was inhibited while the proliferation of HaCat cells was increased. All the difference reached statistical significance (P

Objectives To study the difference in the expression of monocyte chemoattractant protein-1 (MCP-1) and soluble intercellular adhesion molecule-1 (sICAM-1) between vitiliginous and non-vitiliginous patches in patients with stable vitiligo, and to compare the number of peripheral regulatory T cells between vitiligo patients in stable stage and normal controls. Methods Suction blister fluid was collected at 1 to 3 h after the suction. The expression of MCP-1 and sICAM-1 in skin tissue fluid was detected with ELISA. The number of regulatory T cells in the blood of vitiligo patients and controls was detected by flow cytometry. Results There was no significant difference in the number of regulatory T cells between vitiligo patients and normal controls. The expression of MCP-1 and sICAM-1 in skin tissue fluid was significantly higher in vitiliginous skin than that in non-vitiliginous patches in patients with common type vitiligo, while there was no significant difference between the two kinds of patches in patients with segmental type vitiligo. Conclusions The immune function is abnormal in vitiliginous skin of the common type vitiligo patients in stable stage, which might explain the lack of success in transplantation for this disease.

0.25)levels in the two groups.In the group treated with42mJ/cm 2 UVB irradiation followed by the addition of EGCG,the numbers of apoptotic and dead cells and Fas mRNA were decreased,but bcl-2protein was increased.Conclusions Low dosage of UVB irradiation could induce apoptosis of keratinocytes.High dosage of UVB irradiation might result in cell death.EGCG could reduce UVB-induced apoptosis of keratinocytes by increasing bcl-2protein and decreasing Fas mRNA.

Objective To explore the effects of antisense NF-?B p65 oligodeoxynucleotides on UV-induced IL-6 secretion of keratinocytes. Methods NF-?B p65 oligodeoxynucleotides were transfected to a keratinocyte cell line, HaCaT cells, mediated by lipofectamine. The NF-?B p65 protein in the cells was measured by Western blot assay, the mRNA level of NF-?B p65 was detected by RT-PCR, and UV-induced IL-6 level was determined by ELISA pre- and post-transfection and/or UVB irradiation. Results The NF-?B p65 protein expression was significantly increased in UVB (10, 20, 30 mJ/cm2 ) irradiation groups as compared with that of the control groups (P

Objective:To analyze dermoscopic and reflectance confocal microscopic characteristics of acanthosis nigricans, and to assess the value of dermoscopy and reflectance confocal microscopy (RCM) in the auxiliary diagnosis of acanthosis nigricans.Methods:A total of 63 patients with acanthosis nigricans were collected from Department of Dermatology, the Third People′s Hospital of Hangzhou from January 2018 to December 2019, and 5 healthy individuals served as controls. Two lesions on the neck and axilla were examined with dermoscopy and RCM separately in each patient. Biopsies were carried out at the sites evaluated by dermoscopy and RCM in 3 patients, followed by routine histopathological examination.Results:Dermoscopy showed papilloma-like hyperplasia in 126 (100%) lesions, "fat finger" structures in 119 (94.4%) , and "gully" structures in 120 (95.2%) . RCM showed hyperpigmentation in the basal layers, downward extension and twisting of dermal papillary rings and "gully" structures in 126 (100%) lesions, moderately to highly refractive particles in the dermal papillary rings in 87 (69.0%) .Conclusion:Acanthosis nigricans has typical dermoscopic and reflectance confocal microscopic characteristics, which can provide a basis for its diagnosis and differential diagnosis.

Objective To evaluate the effect of green tea polyphenol epigallocatechin-3-gallate (EGCG)and ultraviolet B (UVB) on the expression of aquaporin 3 and epidermal growth factor receptor (EGFR)/extracellular signal-regulated protein kinase (ERK) signaling pathway in keratinocytes.Methods Twenty healthy human subjects were enrolled in this study.Both legs of each subjects were separated into 4 areas to remain untreated (control area),be topically treated with 3％ and 1％ EGCG cream and the vehicle of EGCG cream respectively once a day for 2 weeks followed by the measurement of skin moisture content and transepidermal water loss (TEWL).Cultured keratinocytes were classified into various groups to be irradiated with different doses (10,20 and 30 mJ/cm2) of UVB,or be pretreated with different concentrations of EGCG (10-7,10-6,10-5 mol/L) or EGFR/ERK phosphorylation inhibitors for 1 hour followed by irradiation with UVB of 30 mJ/cm2.After various durations of additional culture,Western blot was conducted to quantify the expression of AQP3 and phosphorylated-EGFR (p-EGFR) and-ERK (p-ERK) of keratinocytes.Data were processed by SPSS 10.0 software,and statistical analysis was carried out by t test.Results Skin moisture content was significantly increased,while TEWL was decreased in healthy skin after treatment with 1％ and 3％ EGCG cream compared with vehicle-treated skin areas and untreated skin areas.Increased AQP3 expression was observed in keratinocytes pretreated with EGCG of 10-7,10-6,10-5 mol/L (172.36 ± 12.42,320.66 ± 15.51 and 368.10 ± 11.39 vs.100.00,t =12.16,26.75 and 38.62 respectively,all P ＜ 0.05) and in those pretreated with the EGFR inhibitor PD153035 of 1.0 μmol/L and ERK inhibitor U0126 of 10 μmol/L (413.85 ± 25.27 and 268.85 ± 16.33 vs.100.00,t =35.16,19.25 respectively,both P ＜ 0.05)compared with those irradiated with UVB of 30 mJ/cm2 alone.UVB irradiation stimulated the phosphorylation of EGFR/ERK in keratinocytes,and the stimulation was markedly inhibited by the pre-treatment with EGCG of 10-7,10-6 and 10-5 mol/L (all P ＜ 0.05).Conclusions EGCG can enhance skin barrier function.AQP3 expression is down-regulated by UVB irradiation in keratinoctyes,while EGCG can inhibit the downregulation likely by suppressing the UVB-induced activation of EGFR and ERK.

ObjectiveTo assess the relationship of interleukin-17(IL-17) and transforming growth factor-β(TGF-β) with the development of vitiligo.MethodsEnzyme-linked immunosorbent assay (ELISA) was carried out to measure the levels of IL-17 and TGF-β in sera from 120 patients with vitiligo and 60 healthy controls.The correlations of serum IL-17 and TGF-β levels with patients' gender,stage and duration of disease,involved body area and presence of family history were assessed.ResultsThe level of serum IL-17 was significantly higher in patients with active vitiligo than in the controls and patients with stable vitiligo (both P ＜0.05).With the rise in involved body area,the level of serum IL-17 gradually increased (x2 =12.656,P ＜0.05).The level of TGF-β in patients with active vitiligo was a little higher than that in the controls and patients with stable vitiligo,with no significant difference between these groups(both P ＞ 0.05).Conclusions The levels of serum IL-17 and TGF-β are somewhat correlated with the activity of vitiligo,and both of them may play a certain role in the pathogenesis of vitiligo.

Objective To investigate the mechanism of repigmentation in vitiligo induced by narrow band ultraviolet B (NB-UVB) via observing the effects of 311 nm NB-UVB on the proliferation of, apoptosis and melanogenesis in melanocytes. Methods Immortalized B10BR melanocytes were irradiated with NB-UVB at varying doses (400, 800 and 1200 mJ/cm2). Subsequently, the proliferation of and apoptosis in melanocytes were detected by MTT assay and flow cytometry, respectively, and the content of melanin was determined by NaOH assay. The expression of BCL-2 mRNA was examined by RT-PCR, and MC-1R expression of melanoeytes by immunocytochemistry and Western blotting. Results The proliferation of and apoptosis in melanocytes experienced no obvious change after irradiation with NB-UVB at the three tested doses. In B10BR melanocytes irradiated with NB-UVB at 400, 800 and 1200 mJ/cm2, the melanin content was 1.42, 1.78, 2.05 times, the mRNA expression of BCL-2 was 1.75, 2.32, 3.28 times, and the protein expression of MC-1R was 1.68, 2.35 and 3.01 times, that in unirradiated melanocytes, respectively. Conclusion NB-UVB irradiation at therapeutic doses could promote the melanogenesis in, enhance anti-oxidative stress activities of melanocytes,by upregulating the expressions of BCL-2 and MC-1R, with no marked effects on the apoptosis in melanocytes.

Objective To investigate the role of activation of Akt/mTOR pathway in denfense against UVB-induced apoptosis in cultured human skin keratinocyte cell line HaCaT. Methods HaCaT cells were irradiated with UVB at different doses for various durations. Western blotting was performed to detect dynamic changes of Akt/mTOR pathway-related signaling molecule, such as phosphorylated-epidermal growth factor receptor (EGFR), -Akt, -4EBP1, etc; apoptosis was estimated by staining with DNA dye Hoechst 33342. To evaluate the role of signaling molecules in defense against UVB-induced apoptosis, HaCaT cells were pretreated before irradiation with EGFR inhibitor (PD153035), PI3K inhibitor (LY294002), mTOR inhibitor (rapamycin) followed by the detection of expressions of signaling molecule and apoptosis. Results UVB could activate Akt/mTOR pathway in a dose- (5 ～ 30 mJ/cm2) and time- (5 ～ 30 min) dependent manner. PD153035,LY 294002 and rapamycin could inhibit UVB-induced activation of the Akt/mTOR pathway. The apoptosis rate in HaCaT cells was upregulated by pretreatment with rapamycin and LY294002. Conclusion The activation of Akt/mTOR signaling pathway could inhibit the UVB-induced apoptosis in cultured HaCaT cells.