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Objective To investigate the effect of three Scutellaria baicalensis extracts (baicalin,baicalein and wogonin) on the apoptosis of a human cutaneous squamous cell carcinoma cell line,SCL-1.Methods Methyl thiazolyl tetrazolium (MTT) assay was performed to detect the proliferation of cultured SCL-1 cells treated with different concentrations (12.5,25,50,75 and 100 μmol/L) of baicalin,baicalein and wogonin for various durations (12,24 and 48 hours).Some SCL-1 cells were treated with baicalin,baicalein and wogonin of 12.5 μ mol/L respectively for 48 hours followed by the detection of cell apoptosis by double staining with annexin V-fluorescein isothiocyanate/propidium iodide in combination with enzyme-linked immunosorbent assay (ELISA),as well as estimation of cell cycle by flow cytometry.The 50％ inhibitory concentration was determined by line regression model and inner insert method,and statistical analysis was carried out by Student's t test,one-way analysis of variance (ANOVA),and Student-Newman-Keuls-q test.Results Baicalin,baicalein and wogonin all inhibited the proliferation of SCL-1 cells in a dose-and time-dependent manner (all P ＜ 0.01).Under the same conditions (treatment concentration and duration),baicalein showed the strongest inhibitory effect on the proliferation of SCL-1 cells,followed by wogonin and baicalin (all P ＜ 0.01).All the Scutellaria baicalensis extracts induced the apoptosis of SCL-1 cells and arrested them in G1-phase.The percentage of cells in G1 phase was 56.37％ ± 2.41％,74.23％ ± 2.02％ and 64.15％ ± 1.87％,and early apoptosis rate was 8.09％ ± 1.02％,24.13％ ± 0.76％ and 14.45％ ± 1.57％,in SCL-1 cells treated with baicalin,baicalein and wogonin of 12.5 μmol/L for 48 hours,respectively,compared to 45.04％ ± 1.93％ and 4.12％ ± 0.29％ in the untreated control cells respectively (F =83.29,186.37,respectively,both P ＜ 0.01).Similarly,there was a downward trend from baicalein to wogonin and baicalin in the effect on cell apoptosis and cell cycle arrest of SCL-1 cells.Conclusions Scutellaria baicalensis can inhibit the growth and induce the apoptosis of SCL-1 cells,which may provide new ideas for the treatment of cutaneous squamous cell carcinoma with traditional Chinese drugs.
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Objective To investigate the in vitro effect of interferon-γ (IFN-γ) and ATRA on the morphological transition, proliferation of and apoptosis in a human cutaneous squamous cell carcinoma cell line SCL-1. Methods Cultured SCL-1 cells were divided into 6 groups to be treated with ATRA of 1 μmol/L, various concentrations ( 100, 500, 1000 U/ml) of IFN-γ, the combination of ATRA of 1 μmol/L and IFN-y of 1000 U/ml,respectively, or to remain untreated. MTT assay and flow cytometry were performed to evaluate the cell proliferation and apoptosis. The morphological features of apoptotic cells were observed by a transmission electron microscope (TEM) and inverted phase contrast microscope after 1% propidium iodide staining. Results IFN-γ could inhibit the proliferation of SCL-1 cells in a dose-dependent manner, and the most pronounced inhibitory effect was observed at a dose of 1000 U/ml . ATRA and IFN-γ induced an apoptosis in SCL-1 cells, and the early apoptosis rate was 4.84%, 11.96% and 18.71% in SCL-1 cells after treated with ATRA of 1 μmol/L, IFN-γ of 1000 U/ml and their combination, respectively. A series of morphological changes characteristic of apoptosis,such as bipolar changes, were observed in SCL-1 cells treated with ATRA and IFN-γ, with the presence of many early apoptotic cells, which showed a trend towards benign differentiation. Conclusions Within a certain concentration range, IFN-γcan promote the differentiation, but inhibit the proliferation of SCL-1 cells in a dose-dependent manner, and ATRA could enhance the effects of IFN-γ.
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Objective To investigate the expression of survivin and bcl-2 in human squamous cell carcinoma (SCC) lesions and cell line SCL-1. Methods Tissue samples from 60 patients with SCC and 10 normal human controls were immunohistochemically stained to detect the expressions of survivin and bcl-2.Western blot was used to measure the expressions of bcl-2 and survivin proteins in HaCaT human keratinocytes and SCL-1 human squamous cell carcinoma cells. Results In normal control tissues, there was no expressions of survivin or bcl-2, while in SCC, the expression rates of bcl-2 and survivin were 70% and 60%, respectively,and there was no statistical correlation between the expressions of bcl-2 and survivin (P >0.05). Neither the expression of survivin nor that of bcl-2 was correlated to patients' age, gender or lesional site (all P >0.05). A statistical correlation was observed between the pathological stage in patients and expression of bcl-2 as well as between lymph node metastasis and expression of survivin (both P < 0.05). Western blot analysis revealed a significant increase in the expression of survivin and bcl-2 in SCL-1 cells compared with HaCaT cells. Con-clusion In SCC, survivin and bcl-2 seem to play their roles via different anti-apoptotic pathways.
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Objective To investigate the influences of UVA on the secretion and expression of chemokine CXCL11/I-TAC by HaCaT cells induced by interferon γ (IFN-γ) and tumor necrosis factor α (TNF-α). Methods HaCaT cells were cultured in the presence of IFN-7 and TNF-a and irradiated with UVA of 2, 4 and 8 J/cm~2, respectively; those cells receiving neither treatment with IFN-γ or TNF-α nor UVA irradiation served as the negative control, and those receiving only cytokine treatment but no irradiation as the positive control. After another 24-hour culture, enzyme-linked immunosorbent assay (ELISA) was performed to detect the protein levels of CXCL11/I-TAC in the supernatant of HaCaT celb, real time PCR to measure the mRNA expression of CXCL11/I-TAC in these HaCaT cells. Results As far as the negative control HaCaT cells were concerned, there was a minor secretion of CXCL11/I-TAC protein and expression of CXCL11/I-TAC mRNA. After treatment with IFN-7 and TNF-a of 10 μg/L, the protein and mRNA expressions of CXCL11/ I-TAC were synergistically upregulated, whereas the induced secretion and expression of CXCL11/I-TAC by HaCaT cells were dose-dependently inhibited by UVA irradiation. Conclusions UVA irradiation inhibits the secretion and expression of CXCL11/I-TAC by HaCaT cells, which in turn suppresses the chemotaxis of Th1/ Tel cells in some degree.
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Objective To investigate the effects of baicalein and acitretin on the apoptosis in a human cutaneous squamous cell carcinoma cell line, SCL-12. Methods Cultured SCL-12 cells were treated with different concentrations of baicalein (3.125, 6.25, 12.5 μmol/L) and acitretin (2.5, 5.0, 10.0 μ mol/L), alone or in combination, for 48 hours. Subsequently, cell proliferation was detected by MTT assay, and cell apoptosis by ELISA as well as annexin V-FITC and propidium iodide double staining. Real-time quantitative RT-PCR was used to detect the expression of Fas mRNA in SCL-12 cells. Results The cell proliferation of SCL-12 cells was inhibited by baicalein and acitretin alone or in combination. The combination of baicalein and acitretin at the three tested concentrations, except for that of baicalein at 3.125 μmol/L and acitretin at 2.5 μmol/L, more strongly inhibited the proliferation of SCL-12 cells compared with baicalein or acitretin alone, and the inhibitory effect was in a dose-dependent manner. The early apoptosis rate was 9.39% ± 1.52%, 20.86% ± 2.16%,36.85% ± 3.26% in SCL-12 cells treated with baicalein of 3.125 μmol/L, acitretin of 5.0 μmol/L alone and their combination, respectively, significantly higher than that in untreated cells (4.39% ± 0.64%, all P <0.05); the induction of apoptosis in SCL-12 cells by the combination of baicalein and acitretin was stronger than that by baicalein or acitretin alone (F = 138.44, P < 0.05). Baicalein and acitretin alone or in combination significantly increased the mRNA expression of Fas in SCL-12 cells, and the effect of their combination was stronger than that of baicalein or acitretin alone. Conclusions Baicalein and aeitretin could inhibit the growth of and induce the apoptosis in SCL-12 cells, and the effect is enhanced by the combination of baicalein and acitretin, which may be associated with the upregulation of Fas expression in SCL-12 cells.
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Objective To observe the efficacy of intense pulsed light(IPL)incorporated with optimal pulse technology (OPT)in the treatment of rosacea.Methods Thiris-two patients with erythematotelangiectatic rosacea and 53 patients with papulopustular rosacea were treated with IPL-OPT for 4 sessions with an interval of 3 weeks.Patients were assessed clinically and photographically by physicians before each treatment.Skin melanin index,erythema index,sebum secretion level and water content in stratum corneum were tested at the baseline,3 weeks after each treatment,and 6 months after the last treatment.Results Three weeks after the last treatment,the effective(more than 60%improvement)rate WaS 81.18%in total,75%in erythematotelangiectatic rosacea,and 84.91%in papulopustular rosacea;there WaS no significant difference between erythematotelangiectatic rosacea and papulopustular rosacea (x2=1.28,P＞0.05).Six months after the last treatment,the total effective rate still remained at 78.82%with no significant difierence from that observed at 3 weeks after the treatment(x2=1.62,P＞0.05).The value of melanin index,erythema index and sebum secretion level decreased significantly after the final treatment,however,water content in stratum corneum remained at the salne level as that before treatment.After 6-month follow-up,no significant change was noticed in the above 4 parameters compared with those obtained at 3 weeks after the treatment.Neither hyperpigmentation nor hypopigmentation was observed during the treatment and follow-up.Conclusion This study demonstrates that IPL-OPT is an effective treatment for rosacea with relatively few side effects.
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Objective To investigate the effects ofthrce generations oftretinoids on the apoptosis of and caspase expressions in human cutaneous squamous cell carcinoma cell line SCL-1. Methods SCL-1 cells were cultured in vitro in the presence of three tretinoins, namely all-trans retinoic acid (ATRA), acitrefln and tazarotene at a concentration of 1×10-5 mol/L. On day 1, 3, 5, MTT assay and ELISA were used to detect the cell proliferation and apoptosis of these SCL-1 cells respectively; on day3, flow cytometry with Annexin V/PI was applied to analyze the cell cycle and early apoptosis, Western blot to measure the protein expressions of caspase-8 and caspase-9 in these cells. Results The growth of SCL-1 cells could be inhibited by ATRA, acitretin or tazarotene of 1×10-5 mol/L in a time-dependent manner (all P<0.01). All the three tretinoids could induce the cell apoptosis of SCL-1 cells (P<0.01), arrest them in G1-phase, and activate caspase-8 and caspase-9 (F=35.50, 25.79, respectively, both P<0.01). Of the three tretinoins, acitretin exerted the strongest effect on all the parameters tested. Conclusions Tretinoins can inhibit the growth and induce the apoptosis of cutaneous squamous carcinoma cells, which may be mediated through Fas- and mitochondria-way.
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Objective To investigate the relationship of CAG repeat length polymorphism in the androgen receptor(AR)gene to the development of acne.Methods A total of 238 patients with ache vulgaris and 207 healthy human controls in Northeast China were included in this study.Genomic DNA was isolated and purified from the blood of these subjects.The CAG repeat lengths in the AR gene were analyzed by somatic microsatellites (STRs).Results A significant difference was found in the CAG repeat number between the male acne Patients(22.70±3.09)and male controls(23.48±2.83,P=0.046),but not between the female cases and controls(23.41±2.87 versus 23.85±0.21.P=0.12).In order to assess the risk associated with CAG repeats,the male and female subjects were dichotomized based on the median repeat length in the corresponding control group as the arbitrary cut-off point.Those men and women with a short CAG repeat length(＜23 in men,and＜24 in women)had a significantly increased risk for agne than those with a long CAG repeat length(men:95%confidence interval,1.21-3.54,OR=2.07,P=0.008;women:95%confidence interval.1.18-3.56,OR=2.05,P=0.01).Conclusions This study strongly indicates that the CAG repeat length in AR gene is associated with the development of acne in Northeast China,and those men with a short CAG repeat length seem to have a high risk for ague.Consequently,CAG repeat length may serve as a genetic susceptibility marker.
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Objective To investigate the molecular transduction mechanisms of apoptosis in cuta-neous squamous cell carcinoma cell line SCL-1 induced by acitretin. Methods SCL-1 cells were cultured and continuously treated with various concentrations of acitretin. Apoptosis was assessed by enzyme-linked immunosorbent assay (ELISA) in these cells on day 1, 3 and 5. Apoptotic cells were observed by acridine orange staining on day 5. The protein expressions of Fas, FasL, Fas-associated death domain (FADD), cas-pase-8, caspase-3 and poly ADP-ribose polymerase (PARP) were examined using Western blot in SCL-1 cells treated with acitretin at 1 x 105 mol/L at different time points. Neutralizing anti-Fas antibody (ZB4,1 μg/mL) was utilized to pretreat SCL-1 cells before the treatment with acitretin, following that, ELISA was done to compare the apoptosis in cells treated with ZB4, acitretin, or the combination of ZB4 and acitretin,respectively. Results Acitretin induced the apoptosis of SCL-1 cells in a dose- and time-dependent manner.Morphologically, acitretin-treated SCL-1 cells showed a typical characteristic of apoptosis. Significant increase in Fas, FasL, and FADD protein expression, as well as the activation of caspase-8, caspase-3 and PARP were induced by the treatment with acitretin. The apoptosis absorbance value was 0.78 ± 0.04 in cells treated with acitretin alone, decreased to 0.41 ± 0.03 in cells treated with ZB4 and acitretin (P ＜ 0.05), .suggesting that ZB4 could block the apoptosis of SCL-1 cells inducedby acitretin. Conclusion Acitretin could induce the apoptosis of cutaneous squamous cell carcinoma cells likely by Fas signaling pathway.
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Objective To investigate the role of immunophenotyping in distinguishing mycosis fun- goides (MF) from lichen planus and psoriasis.Methods The expression of CD1a,CD4,CD8,ICAM-1, LFA-1,HLA-DR,CD30 and CD7 was measured by ABC immunohistochemical technique in specimens ob- tained from lesional skin of 15 cases of MF,17 cases of lichen planus and 17 cases of psoriasis,and in the skin of 6 healthy controls.Results In the lesional epidermis of MF,the density of cells positive for CD1a, CD30 or ICAM-1,was significantly higher (mononuclear cells,P＜0.001;dendritic cells,P＜0.01) than that in the lesional epidermis of lichen planus,psoriasis and in the skin of healthy controls.The density of cells positive for CD4 or CD8 and of dendritic cells positive for HLA-DR was higher in lesional epidermis of MF than in that of lichen planus.The linear density of CD1a-positive cells (P＜0.01),the percentages of cells positive for ICAM-1 (P＜0.05) or LFA-1 (P＜0.05) were all higher in the lesional dermis of MF than in that of lichen planus.As far as the CD7-positive cell density was concerned,it was higher in the lesional dermis of lichen planus and psoriasis than in that of MF and skin of healthy controls (P＜0.01), while no difference was found between the epidermis of MF and that of lichen planus or psoriasis.Conclu- sion There are differences in the expression of CD1a,CD4,CD8,ICAM-1,LFA-1,HLA-DR,CD30 and CD7 in the lesional skin of MF,lichen planus and psoriasis,which may provide a clue to the pathogenesis of these diseases.
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Objective To investigate the role of insulin resistance in the pathogenesis of childhood benign acanthosis nigricans with obesity and the relationship between insulin resistance and insulin receptors.Methods Fasting plasma insulin and the amount of erythrocyte insulin receptors were determined among17children with benign acanthosis nigricans with obesity(CBANO),with16cases of simple adiposis and15healthy children as controls.Results The levels of plasma insulin were higher in CBANO than those in sim-ple adiposis and healthy controls;however,the amount of erythrocyte insulin receptors was lower in CBANO than that in simple adiposis and healthy controls.Conclusion Insulin resistance caused by diminished in-sulin receptors may play an important role in pathogenesis of CBANO.
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Objective To investigate the distributions of haptoglobin (Hp) phenotypes and their clinical significance in five skin diseases. Methods Haptoglobin phenotypes were determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) in a discontinuous buffer system, and then confirmed by staining with silver nitrate. Results The distribution of Hp1-1, Hp2-1, Hp 2-2 of Han ethnic group was 9.9 percent, 46.9 percent, 43.2 percent, respectively. Compared with normal controls, the frequency of Hp2-2 in SLE group (59.3%) was markedly high, especially in those with kidney damage, that of Hp2-1 in psoriasis(30.9%) was low, and that of Hp1-1 in eczema was high, especially in those without exudation (27.4%). There was no statistical significance for the distributions of Hp phenotypes in secondary syphilis and condyloma acuminatum. Conclusion Hp phenotypes might be associated with the pathogensis and some clinical manifestations in SLE, psoriasis of eczema.
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Objective To investigate the expression of haptoglobin(Hp)mRNA and protein in nor-mal human epidermal cells and human keratinocyte cell line-HaCaT cells.Methods In situ hybridization and RT-PCR were used to detect the expression of Hp mRNA in normal human epidermal cells and HaCaT cells.Immunohistochemistry was used to detect the expression of Hp on normal human epidermal cells.Im-munohistochemistry and Western blot were used to detect Hp expression on HaCaT cells.Results There was Hp mRNA expression in normal human keratinocytes and HaCaT cells.There was no Hp mRNA expres-sion in normal human epidermal Langerhans cells.There were some Hp positive dendritic cells in normal human epidermis.There was no obvious Hp protein staining in the HaCaT cells by immunohistochemistry.There was Hp protein band from HaCaT cells by Western blot.Conclusions There is Hp mRNA expression in normal human keratinocytes and HaCaT cells which suggests that normal human keratinocytes and HaCaT cells have the ability to synthesize Hp protein.Normal human epidermal Langerhans cells have no ability to synthesize Hp protein.There is small amount of Hp protein in HaCaT cells.
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Objective To detect human papillomavirus(HPV)DNA in peripheral blood of patients with condyloma acuminata(CA).Methods Polymerase chain reaction(PCR)was used to amplify the HPV DNA in skin lesions and peripheral blood of30patients with CA,and20normal blood donors as controls.Re-striction endonucleases Rsa I and Pst I were used for HPV DNA typing.The PCR products from1patient who was HPV positive for both skin lesions and peripheral blood,were cloned and sequenced.Results HPV DNA was identified in all CA skin lesions.HPV DNA was identified in peripheral blood mononuclear cells(PBM-Cs)from8of30(26.7%)patients and in none of20control subjects(P
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Objective To analyze the possible association of psoriasis vulgaris with herpes simplex virus type 1 (HSV-1). Methods Polymerase chain reaction was used to detect HSV-1 DNA in lesional skin biopsies, periphery blood mononuclear cells (PBMCs)and throat swabs from patients with psoriasis vulgaris, and ELISA was used to detect IgM and IgG antibodies against HSV-1 in sera from these patients. Results The positive detection rates of HSV 1 DNA in lesional skin biopsies, PBMCs and throat swabs were 37.5%, 18.6%and 18.8%, respectively. Anti HSV 1 IgM and IgG antibodies were positive in 37.2%and 53.5%of serum specimens, respectively. The detection rates of HSV 1 DNA in lesional skin biopsies and PBMCs, and IgM antibody in sera were significantly higher than those in normal controls. In psoriatic patients of guttate type the positive detection rates of HSV 1 DNA and IgM antibody were significantly higher than those in the plaque type. Conclusions There is strong association of psoriasis vulgaris, especially the guttate type, with HSV 1, and there may be recent infection of HSV 1 in these patients.