IL-23/IL-17 immune axis mediates the imiquimod-induced psoriatic inflammation by activating ACT1/TRAF6/TAK1/NF-κB pathway in macrophages and keratinocytes.

The interleukin-23 (IL-23)/IL-17 immune axis has been linked to the pathology of psoriasis, but how this axis contributes to skin inflammation in this disease remains unclear. We measured inflammatory cytokines associated with the IL-23/IL-17 immune axis in the serum of patients with psoriasis using enzyme-linked immunosorbent assays. Psoriasis was induced in male C57BL/6J mice using imiquimod (IMQ) cream, and animals received intraperitoneal injections of recombinant mouse anti-IL-23A or anti-IL-17A antibodies for 7 days. The potential effects of the IL-23/IL-17 immune axis on skin inflammation were assessed based on pathology scoring, hematoxylin-eosin staining of skin samples, and quantitation of inflammatory cytokines. Western blotting was used to evaluate levels of the following factors in skin: ACT1, TRAF6, TAK1, NF-κB, and pNF-κB. The serum of psoriasis patients showed elevated levels of several cytokines involved in the IL-23/IL-17 immune axis: IL-2, IL-4, IL-8, IL-12, IL-17, IL-22, IL-23, and interferon-γ. Levels of IL-23p19 and IL-17 were increased in serum and skin of IMQ-treated mice, while ACT1, TRAF6, TAK1, NF-κB, and pNF-κB were upregulated in the skin. A large proportion of NF-κB p65 localized in nucleus of involucrin+ cells in the epidermis and in F4/80+ cells of the dermis of psoriatic lesional skin. Treating these animals with anti-IL-23 or anti-IL-17 antibodies improved pathological score and immune imbalance, mitigated skin inflammation and downregulated ACT1, TRAF6, TAK1, NF-κB, and pNF-κB in skin. Our results suggest that skin inflammation mediated by the IL-23/IL-17 immune axis in psoriasis involves activation of the ACT1/TRAF6/TAK1/NF-κB pathway in keratinocytes and macrophage.

MiR-149-5p inhibits cell proliferation, promotes cell apoptosis and retards cell cycle of IL-22-stimulated HaCaT and NHEK keratinocytes via regulating PDE4D.

BACKGROUND: Psoriasis is a chronic autoimmune skin disease with unclear pathogenesis. It was found that miR-149-5p was significantly decreased in psoriatic lesion tissues. In this study, we aims to investigate the role and related molecular mechanism of miR-149-5p on psoriasis. METHOD: IL-22 was used to stimulate HaCaT and NHEK cells to establish psoriasis model in vitro. The miR-149-5p and phosphodiesterase 4D (PDE4D) expression levels were detected by quantitative real-time PCR. HaCaT and NHEK cells proliferation was determined by Cell Couting Kit-8 assay. The cell apoptosis and cell cycle were detected by flow cytometry. The cleaved Caspase-3, Bax and Bcl-2 protein expressions were detected by western blot. The targeting relationship between PDE4D and miR-149-5p was predicted and confirmed by Starbase V2.0 and dual-luciferase reporter assay, respectively. RESULT: There was a low expression level of miR-149-5p and a high expression of PDE4D in psoriatic lesion tissues. MiR-149-5p could target PDE4D. IL-22 promoted HaCaT and NHEK cells proliferation, while inhibited cell apoptosis and accelerated cell cycle. Moreover, IL-22 decreased the expressions of cleaved Caspase-3 and Bax, and increased the expression of Bcl-2. And the overexpressed miR-149-5p promoted HaCaT and NHEK cells apoptosis, inhibited cell proliferation and retarded cell cycle, meanwhile increased the cleaved Caspase-3 and Bax expressions, decreased the Bcl-2 expression. In addition, PDE4D overexpression has the opposite effect as miR-149-5p. CONCLUSION: The overexpressed miR-149-5p inhibits IL-22-stimulated HaCaT and NHEK keratinocytes proliferation, promotes cell apoptosis and retards cell cycle by down-regulating the expression of PDE4D, which could be the promising therapeutic target of psoriasis.

Alkannin exerts antitumor properties in cutaneous squamous cell carcinoma by inducing apoptosis and shifting the M1/M2 polarization of tumor-associated macrophages by upregulating PTEN.

Cutaneous squamous cell carcinoma (CSCC) is a common cancer in humans and is the second major type of skin cancer that causes death in humans. In this article, we investigated the effects of alkannin on CSCC progression. We revealed that alkannin curbed CSCC cell viability in a dose-dependent manner and accelerated CSCC cell apoptosis. In addition, alkannin expedited macrophage M1 polarization while curbing M2 polarization. Moreover, alkannin elevated phosphatase and tensin homolog (PTEN) abundance in CSCC cells. The results of bioinformatics analysis revealed that alkannin might modulate CSCC via PTEN. Downregulation of PTEN reversed the effects of alkannin on apoptosis of CSCC cells and M1/M2 polarization of macrophages. Alkannin reduced CSCC tumor growth in a mouse xenograft model. In conclusion, alkannin curbed the advancement of CSCC by expediting apoptosis and facilitating M1 polarization of macrophages by upregulating PTEN. These data may offer a therapeutic approach against CSCC.

The effect of Xiaobi decoction on minimal erythema dose of irradiation with 308 nm excimer lamp in guinea pigs.

BACKGROUND: Psoriasis is a chronic autoimmune-mediated inflammatory skin disease. The main clinical manifestation of this complex condition is scaly erythema. Excimer 308-nm light can be used to selectively and safely treat skin lesions. In clinical practice, the combination of Xiaobi decoction combined with 308-nm excimer light therapy has been found to have a beneficial effect on advanced psoriasis vulgaris. However, the effect of Xiaobi decoction on light sensitivity in psoriasis patients has not been explored. Therefore, this study aimed to observe the effect of Xiaobi decoction on the minimal erythema dose (MED) value in guinea pigs. METHODS: Eighty guinea pigs were divided into a Xiaobi decoction group and a control group according to the serial number: guinea pigs with odd numbers were assigned into the Xiaobi decoction group, and those with even numbers were assigned into the control group. The Xiaobi decoction group was administrated Xiaobi decoction by gastric lavage, while the control group was given distilled water by gastric lavage. The back skin of the guinea pigs in the two groups was irradiated with a 308-nm excimer lamp before gavaging, and after 10, 20, and 30 days of gavaging. The MED values 24 hours after irradiation were recorded. RESULTS: The average MED in the Xiaobi decoction group was (800±126.5) mJ/cm2, compared with (780±107.7) mJ/cm2 in the control group, and the difference was not significant (P>0.05). After 10 days, 20 days, and 30 days of gavaging, the MED values of the guinea pigs in the Xiaobi decoction group were significantly lower than those of the control group (P<0.05). Blood tests showed that the levels of vascular endothelial growth factor (164.5±25.7 vs. 145.3±27.4, P=0.002) and interleukin-23 (1.8±0.7 vs. 1.5±0.5, P=0.030) were significantly lower in the Xiaobi decoction group than in the control group. CONCLUSIONS: Xiaobi decoction can reduce the MED value of guinea pigs, and increase the sensitivity of skin lesions to 308-nm excimer light and the amount of light absorbed by skin lesions. These results may represent the mechanisms of action of Xiaobi decoction in the treatment of psoriasis.

Long non-coding RNA SCARNA2 induces cutaneous squamous cell carcinoma progression via modulating miR-342-3p expression.

BACKGROUND: Long non-coding RNAs (lncRNAs) play important roles in the progression of tumors. However, the function and expression of SCARNA2 in cutaneous squamous cell carcinoma (cSCC) is still unreported. METHODS: A quantitative polymerase chain reaction was applied to study the expression of SCARNA2 and miR-342-3p. Cell counting kit-8, flow cytometry and transwell assays were performed to study cell growth, cycle and cell invasion. RESULTS: We found that SCARNA2 expression is up-regulated in cSCC cell lines and SCARNA2 expression is higher in cSCC tissues than in adjacent non-tumor specimens. Ectopic expression of SCARNA2 promoted cell growth, cell cycle and invasion in SCC13 cells. In addition, the data indicate that miR-342-3p expression is down-regulated in cSCC cell lines and miR-342-3p is down-regulated in cSCC tissues compared to adjacent non-tumor specimens. We showed that the SCARNA2 expression is negatively associated with miR-342-3p in cSCC. Moreover, we noted that SCARNA2 sponges miR-342-3p expression in cSCC cells. Overexpression of SCARNA2 suppressed the miR-342-3p expressed in SCC13 cells. We found that elevated expression of SCARNA2 promotes cell growth, cell cycle and invasion via regulating miR-342-3p expression in SCC13 cells. CONCLUSIONS: These data suggest that SCARNA2 acts in an oncogenic role and may be a potential target for cSCC.

Neogambogic acid induces apoptosis of melanoma B16 cells via the PI3K/Akt/mTOR signaling pathway.

BACKGROUND: Neogambogic acid, as one of the main components of gamboge, exhibits high activities against various tumors. OBJECTIVE: To explore the mechanism by which melanoma B16 cell apoptosis was induced by neogambogic acid. METHODS: Melanoma B16 cells were treated with different concentrations of neogambogic acid solutions (0, 1.5, 3.0, 6.0 µM). The proliferation inhibition rate was measured by MTT assay. Cell morphology was observed by inverted microscope. Cell migration and invasion were tested by Transwell assay. Flow cytometry was performed to detect the apoptosis rate and cell cycle of B16 cells. The expressions of PI3K/Akt/mTOR signaling pathway-related proteins were detected by Western blot. RESULTS: The proliferation inhibition rate of B16 cells significantly increased with rising neogambogic acid concentration (P<0.05). The invasive and migration capacities of B16 cells decreased significantly after treatment with neogambogic acid (P<0.05). The apoptosis rate also increased with rising concentration of neogambogic acid. After 24 h of treatment, the percentage of G0/G1 phase cells increased gradually as the neogambogic acid concentration rose, whereas those of S phase and G2/M phase cells decreased. With increasing concentration of neogambogic acid, the expressions of p-PI3K, p-Akt and p-mTOR proteins reduced in a time-dependent manner, but those of PI3K and Akt proteins remained basically unchanged. CONCLUSION: Neogambogic acid can inhibit the proliferation, invasion and migration of melanoma B16 cells and induce their apoptosis, which may be regulated via the PI3K/Akt/mTOR signaling pathway.

Diagnostic utility of QuantiFERON-TB Gold In-Tube test in pediatric tuberculosis disease in Taiwanese children.

PURPOSE: To compare the performance of a commercial interferon-gamma release assay, QuantiFERON TB Gold-in-Tube (QFG-IT) with the tuberculin skin test (TST) in Taiwanese children for the diagnosis of active tuberculosis (TB). METHODS: A retrospective chart analysis of pediatric patients (<18 years of age) who underwent QFG-IT tests and TST for the confirmation of active TB between January 2008 and June 2014. RESULTS: The sensitivity of QFG-IT was 100% [95% confidence interval (CI): 63.1-100], versus sensitivity of 62.5% for TST (95% CI 24.5-91.5). The positive predictive value of QFG-IT was 100 (95% CI: 89.7-100), while the negative predictive value for TST was 86.9% (95% CI: 67-96.3). Among three patients with Bacillus Calmette-Guérin (BCG) osteitis, two patients with TST were positive, but all tested samples for QFG-IT were negative. CONCLUSION: QFG-IT assay was more sensitive for the diagnosis of TB disease than TST in an intermediate burden population with universal neonatal BCG vaccination. The increased recognition of BCG induced osteitis in recent years has alerted physicians that BCG induced lesions should be suspected when TST is positive but QFG-IT is negative. Despite higher costs for QFG-IT than TST, they have additional value for the diagnosis of active TB and should be performed when a diagnosis of TB remains in doubt.

Impact of HLA-B*58:01 allele and allopurinol-induced cutaneous adverse drug reactions: evidence from 21 pharmacogenetic studies.

Allopurinol is widely used for hyperuricemia and gouty arthritis, but is associated with cutaneous adverse drug reactions (CADRs). Recently, HLA-B*58:01 allele was identified as a strong genetic marker for allopurinol-induced CADRs in Han Chinese. However, the magnitude of association and diagnosis value of HLA-B*58:01 in allopurinol-induced CADRs remain inconclusive. To investigate this inconsistency, we conducted a meta-analysis of 21 pharmacogenetic studies, including 551 patients with allopurinol-induced CADRs, and 2,370 allopurinol-tolerant controls as well as 9,592 healthy volunteers. The summary OR for allopurinol-induced CADRs among HLA-B*58:01 carriers was 82.77 (95% CI: 41.63 - 164.58, P < 10-5) and 100.87 (95% CI: 63.91 - 159.21, P < 10-5) in matched and population based studies, respectively. Significant results were also observed when stratified by outcomes and ethnicity. Furthermore, the summary estimates for quantitative analysis of HLA-B*58:01 allele carriers in allopurinol-induced CADRs screening were as follows: sensitivity, 0.93 (95% CI: 0.85 - 0.97); specificity, 0.89 (95% CI: 0.87 - 0.91); positive likelihood ratio, 8.24 (95% CI: 6.92 - 9.81); negative likelihood ratio, 0.084 (95% CI: 0.039 - 0.179); and diagnostic odds ratio, 98.59 (95% CI: 43.31 - 224.41). The AUSROC was 0.92 (95% CI: 0.89-0.94), indicating the high diagnostic performance. Our results indicated that allopurinol-SCAR is strongly associated with HLA-B*58:01, and HLA-B*58:01 is a highly specific and effective genetic marker for the detection allopurinol-induced CADRs, especially for Asian descents.