Leptin induces altered differentiation of keratinocytes by inducing insulin resistance: implications for metabolic syndrome-induced resistance of psoriatic therapy.

Background: Psoriatic patients tend to develop metabolic syndrome (MS). MS accelerates psoriasis, but the exact molecular mechanisms are poorly understood.Objectives: We aim to investigate the impact of leptin on keratinocyte insulin sensitivity and explore its underlying molecular mechanism, which might play a role in the pathogenesis of this disease.Methods: ELISA and immunohistochemistry were applied respectively to detect the level of leptin in serum and in lesion of psoriatic patients with and without MS. The HaCaT cell line was cultured and western-blot assay was performed to assess the change of insulin sensibility. q-PCR and western-blot assay were applied to detect the SOCS3 expressions. Knockdown of SOCS3 were generated in HaCaT cell line by siRNA. Leptin and insulin were treated for 6 days and K10 expression was evaluated by western-blot assay.Results: Patients with MS had higher level of leptin in serum and lesions than their counterparts without MS. Serum levels of leptin was negatively correlated to PASI decline index in psoriatic patients. Long-term treatment of leptin induced insulin resistance in HaCaT cell line, as indicated by elevated expression of p-IRS-1 (ser636) and lower p-PKB (ser473). Leptin treatment up-regulated the mRNA and protein expression of SOCS3. Knockdown of SOCS3 blocked the effect of leptin-induced insulin resistance. Leptin treatment attenuated insulin-elicited K10 expression.Conclusions: Leptin induces insulin resistance by upregulating SOCS3 and give rise to differentiation disorder of keratinocyte. Insulin resistance may serve as a target for anti-psoriatic therapies.

Preparation and partial pharmacodynamic studies of luliconazole ethosomes.

As a drug carrier, ethosome is found to be efficient in delivering drug to the deep skin layers through stratum corneum, and the purpose of this paper is to develop luridazole ethosomes acting as an optimal choice for transdermal antifungal drugs. The luliconazole ethosomes were prepared by thin-film hydration, and evaluated for morphology, size, entrapment efficiency (EE), stability and deformability. In vitro, the transdermal experiment was performed on excised rat skin by Franz diffusion cell, and minimum inhibitory concentration (MIC) was applied to determine antifungal activity. In vivo, the irritation of luliconazole ethosomes was also observed in rats. The luliconazole ethosomes were prepared with 5% (w/v) lecithin, 45% (v/v) ethanol and 8-min ultrasound, and characterised with small and uniform particle size, high EE of about 70%. These ethosomes possessed good deformability, were stable and affected by light and high temperature. The cumulative amount permeated of different dosage forms at 48 h from high to low was: ethosome > ointment > liposome > hydroalcoholic solution (p < 0.05), and the sum of the luliconazole retention of skin from high to low at 48 h was: ethosome/ointment > liposome > hydroalcoholic solution (p < 0.05). In the antifungal experiment, the MICs from high to low were: hydroalcoholic solution > liposome > ethosome (p < 0.05), and Trichoderma was more sensitive to luliconazole than Candida. There was no skin irritation observed after treatment of luliconazole ethosomes. The luliconazole ethosomes are firstly prepared in our study, which have little stimulation, better permeation effect and antifungal activity, offering a new perspective for choosing clinical antifungal drugs in the Department of Dermatology.

Photodynamic Therapy for Genital Warts Causes Activation of Local Immunity.

BACKGROUND: 5-aminolevulinic acid photodynamic therapy (PDT) for genital warts is effective, safe, and can prevent recurrence. It is believed that PDT can induce immune responses, but the mechanism is not completely understood. OBJECTIVES: The objectives of this article are to confirm the effect of PDT for genital warts on local immunity and to investigate the recruitment and significance of immune cells in tissues. METHODS: Local immune changes in T lymphocytes (CD3+, CD4+, CD8+), plasmacytoid dendritic cells (pDCs) (CD123+), and myeloid dendritic cells (CD1a+) after PDT in patients were evaluated by immunohistochemistry staining. Changes in mRNA levels of IFN-γ, IFN-α, IFN-ß, interferon-stimulated gene 15 kDa (ISG-15), Mx2, Toll-like receptor 9 (TLR9), and interferon regulatory factor 7 (IRF7) were analyzed by real-time quantitative polymerase chain reaction. RESULTS: At 4 hours after PDT, CD4+ increased, accompanied by increased levels of mRNA expression of IFN-γ, but CD4+ and mRNA expression levels of IFN-γ were decreased at 24 hours after PDT. CD123+ pDCs showed an increasing trend. CD1a+ LCs in the epidermis gradually decreased, and DCs in the epidermis gradually increased. CD3+ infiltrated and migrated to the superficial dermis, but CD8+ did not change significantly after PDT. The mRNA expression levels of IFN-α, IFN-ß, ISG-15, Mx2, TLR9, and IRF7 showed an increasing trend after PDT. As compared with the patients without significantly increased IFN-α and IFN-ß after PDT sessions, patients with significant increases needed fewer sessions of PDT for remission. CONCLUSIONS: PDT for genital warts can activate T lymphocyte-mediated, DC-related, and pDC-related immunity. The clinical efficacy of PDT for genital warts may be related to the increased levels of IFN-α and IFN-ß after treatment.

Safety and efficacy of etanercept monotherapy for moderate-to-severe plaque psoriasis: A prospective 12-week follow-up study.

Etanercept has been shown to be effective for the treatment of moderate-to-severe plaque psoriasis. Since most clinical trials examined etanercept in combination with other drugs, the efficacy and safety of etanercept monotherapy for moderate-to-severe plaque psoriasis have not been well established. This prospective study enrolled 61 Chinese patients with moderate-to-severe plaque psoriasis to explore the efficacy and safety of etanercept monotherapy. These patients were treated with etanercept at a subcutaneous dose of 25 mg, twice a week, for 12 weeks. All the 61 patients completed the treatment and showed significant improvement in psoriasis area and severity index (PASI) scores. At 4, 8, and 12 weeks after treatment, the response rates (PASI75) were 0%, 21.31%, and 40.98%, respectively. It was concluded that etanercept monotherapy is efficacious and safe for patients with moderate- to-severe plaque psoriasis.

Safety and Efficacy of Etanercept Monotherapy for Moderate-to-severe Plaque Psoriasis: A Prospective 12-week Follow-up Study / 华中科技大学学报（医学）（英德文版）

Etanercept has been shown to be effective for the treatment of moderate-to-severe plaque psoriasis.Since most clinical trials examined etanercept in combination with other drugs,the efficacy and safety of etanercept monotherapy for moderate-to-severe plaque psoriasis have not been well established.This prospective study enrolled 61 Chinese patients with moderate-to-severe plaque psoriasis to explore the efficacy and safety of etanercept monotherapy.These patients were treated with etanercept at a subcutaneous dose of 25 mg,twice a week,for 12 weeks.All the 61 patients completed the treatment and showed significant improvement in psoriasis area and severity index (PASI) scores.At 4,8,and 12 weeks after treatment,the response rates (PASI75) were 0％,21.31％,and 40.98％,respectively.It was concluded that etanercept monotherapy is efficacious and safe for patients with moderate-to-severe plaque psoriasis.

B16 cell lysates plus polyinosinic-cytidylic acid effectively eradicate melanoma in a mouse model by acting as a prophylactic vaccine.

Th1 antigen-specific T cells secrete interferon-γ, which is able to kill antigen-specific cancer cells and is helpful for cancer vaccines. The aim of the present study was to explore whether B16 cell lysates plus polyinosinic-cytidylic acid (poly I:C) can effectively inhibit the progression of melanoma in an animal model. In the present study, C57BL/6 mice were divided into three groups, with each group containing more than six mice. The groups of mice were immunized twice with B16 cell lysates plus poly I:C, B16 cell lysates, or phosphate-buffered saline only, respectively. The in vivo results demonstrated that splenocytes from the mice immunized with B16 cell lysates plus poly I:C contained higher percentages of CD3+CD8+ T lymphocytes and CD3+CD4+ T lymphocytes, which were detected by a fluorescence-activated cell sorter, and produced higher levels of antigen-specific splenocyte proliferation activity, as detected by MTT assay. The splenocytes from the mice immunized with B16 cell lysates in combination with poly I:C produced higher levels of interferon­Î³, as detected by quantitative polymerase chain reaction and ELISA, as well as cytotoxic T lymphocyte activity when stimulated in vitro with B16 lysates. Additionally, subcutaneous immunization of the C57BL/6 mice with B16 cell lysates plus poly I:C conferred greater protection against tumor-forming B16 melanoma cells than that of the mice immunized with injection of B16 cell lysate alone. In conclusion, the cancer vaccine of B16 cell lysates plus poly I:C exerts potently protective effects that polarize responses toward Th1 and elicit antitumor immunity.

Inhibitory effect of melanoma differentiation associated gene-7/interleukin-24 on invasion in vitro of human melanoma cancer cells.

The acquisition of metastasis potential is a critical point for malignant tumors. Melanoma differentiation associated gene-7/interleukin-24 (mda-7/IL-24) is a potential tumor suppress gene and frequently down-regulated in malignant tumors. It has been implicated that overexpression of MDA-7 led to proliferation inhibition in many types of human tumor. Invasion is an important process which is potential to promote tumor metastasis. However, the role and potential molecular mechanism of mda-7/IL-24 to inhibit the invasion of human melanoma cancer is not fully clear. In this report, we identified a solid role for mda-7/IL-24 in invasion inhibition of human melanoma cancer LiBr cells, including decreasing of adhesion and invasion in vitro, blocking cell cycle, down-regulating the expression of ICAM-1, MMP-2/9, CDK1, the phosphorylation of ERK and Akt, NF-κB and AP-1 transcription activity. Meanwhile, there was an increased expression of PTEN in mda-7/IL-24 over-expression LiBr cells. Our results demonstrated that mda-7/IL-24 is a potential invasion suppress gene, which inhibits the invasion of LiBr cells by the down-regulation of ICAM-1, MMP-2/9, PTEN, and CDK1 expression. The molecular pathways involved were the MAPK/ERK, PI3K-Akt, NF-κB, and AP-1. These findings suggest that mda-7/IL-24 may be used as a possible therapeutic strategy for human melanoma cancer.

Aqueous extracts of Ocimum grasstimum inhibits lipopolysaccharide-induced interleukin-6 and interleukin-8 expression in airway epithelial cell BEAS-2B.

OBJECTIVE: To investigate the antiinflammatory activities of aqueous extract of Occimum gratissmium (OGE) with emphasis on expression of proinflammatory cytokines in Lipopolysaccharide (LPS)-stimulated epithelial cell BEAS-2B. METHODS: Effects of OGE on cell viability were determined by MTT assay. mRNA expression were analyzed by and reverse transcription polymerase chain reaction (RT-PCR) and quantitative real-time PCR. Activation of kinase cascades was investigated by immunoblot. Intracellular reactive oxygen species (ROS) was analyzed by flow cytometry. RESULTS: OGE (<200 µg/mL) treatment or pretreatment and following LPS exposure slightly affected viability of BEAS-2B cells. Increase of interleukin (IL)-6 and IL-8 and the elevated level of intracellular ROS in LPS-stimulated BEAS-2B cells were diminished by OGE pretreatment in a dose-dependent manner. OGE suppressed inflammatory response-associated mitogen-activated protein kinases (MAPKs) and Akt activation. Additionally, OGE pretreatment increased level of cellular inhibitor of κBα (IκBα) and inhibited nuclear translocation of nuclear factor kappa B (NF-κB). CONCLUSION: These findings indicate that significant suppression of IL-6 and IL-8 expressions in LPS-stimulated BEAS-2B cells by OGE may be attributed to inhibiting activation of MAPKs and Akt and consequently suppressing nuclear translocation of NF-κB.

Facile synthesis of water-soluble and size-homogeneous cadmium selenide nanoparticles and their application as a long-wavelength fluorescent probe for detection of Hg(II) in aqueous solution.

Highly luminescent uncoated water-soluble and mono-disperse CdSe nanoparticles (NPs) have been prepared facilely. Uncoated CdSe core NPs possessing a good size distribution was accompanied with long wavelength of fluorescence emission. It is interesting to note that these functionalized NPs are soluble in water medium stably for more than 1 month, and no significant changes were found in the optical characteristics in comparison with fresh CdSe NPs prepared. The functionalized CdSe NPs exhibited strong specific affinity for mercury(II) through their surface functional groups. Based on the significant quenching of fluorescence emission of functionalized CdSe NPs with a long-wavelength 630nm, a simple, rapid and specific detection for Hg(II) was proposed. Under optimum conditions, the response of linearly proportional to the concentration of Hg(II) is between 0mol/L and 1.25x10(-6)mol/L, and the limit of detection is 4.50x10(-9)mol/L. The relative standard deviation (R.S.D.) of six replicate measurements is 2.0% for 2.0x10(-7)mol/L of Hg(II). In terms of fluorescence quenching at 630nm of CdSe NPs, no obvious wavelength shift or no new emission band in presence of Hg(II) at pH 7.50 of phosphate buffer solution were found; furthermore, a significant reduction in absorbance at 230nm of CdSe NPs was first observed in our work. We could speculate that Hg(II) as an effective quencher (even at low concentration) for functionalized CdSe NPs emission suggests that it is capable of directly intercepting one of the charge carriers, thus disrupting the recombination process.