Interleukin-4 affects microglial autophagic flux

Interleukin-4 plays an important protective role in Alzheimer’s disease by regulating microglial phenotype,phagocytosis of amyloid-β,and secretion of anti-inflammatory and neurotrophic cytokines.Recently,increasing evidence has suggested that autophagy regulates innate immunity by affecting M1/M2 polarization of microglia/macrophages.However,the role of interleukin-4 in microglial autophagy is unknown.In view of this,BV2 microglia were treated with 0,10,20 or 50 ng/mL interleukin-4 for 24,48,or 72 hours.Subsequently,light chain 3-II and p62 protein expression levels were detected by western blot assay.BV2 microglia were incubated with interleukin-4(20 ng/mL,experimental group),3-methyladenine(500μM,autophagy inhibitor,negative control group),rapamycin(100 nM,autophagy inductor,positive control group),3-methyladenine+interleukin-4(rescue group),or without treatment for 24 hours,and then exposed to amyloid-β(1μM,model group)or vehicle control(control)for 24 hours.LC3-II and p62 protein expression levels were again detected by western blot assay.In addition,expression levels of multiple markers of M1 and M2 phenotype were assessed by real-time fluorescence quantitative polymerase chain reaction,while intracellular and supernatant amyloid-βprotein levels were measured by enzyme-linked immunosorbent assay.Our results showed that interleukin-4 induced microglial autophagic flux,most significantly at 20 ng/mL for 48 hours.Interleukin-4 pretreated microglia inhibited blockade of amyloid-β-induced autophagic flux,and promoted amyloid-βuptake and degradation partly through autophagic flux,but inhibited switching of amyloid-β-induced M1 phenotype independent on autophagic flux.These results indicate that interleukin-4 pretreated microglia increases uptake and degradation of amyloid-βin a process partly mediated by autophagy,which may play a protective role against Alzheimer’s disease.

Local hyperthermia cleared multiple cutaneous warts on a nephrotic syndrome patient

Cutaneous warts are caused by human papillomavirus infection. Immunosuppressive state is one of the risk factors of human papillomavirus infection. A girl diagnosed of nephrotic syndrome and on immunosuppressive therapy developed multiple common warts. We treated her on a single lesion by local hyperthermia therapy at 44 ℃ for 3 consecutive days, each therapy lasted for 30 min. Ten days later, the patient received another 2 consecutive therapy. All lesions are completely resolved at the 9th week after the treatment. No recurrent sign was observed in a 3-mo follow-up. Side effects included burning sensation, stabbing pain at the target site during treatment.

Interleukin-18 exacerbates skin inflammation and affects microabscesses and scale formation in a mouse model of imiquimod-induced psoriasis

Background: As a potent pro-inflammatory cytokine of the interleukin (IL)-1 family, IL-18 was elevated in early active progressive plaque-type psoriatic lesions and that serum or plasma levels of IL-18 correlated with the Psoriasis Area and Severity Index (PASI). Although results from previous studies have established that IL-18 may aggravate psoriatic inflammation, the mechanisms of this process remain unknown. In this study, IL-18 knock out (KO) mice and wild-type (WT) mice were used to investigate the effects of IL-18 within a mouse model of psoriasis. Methods: WT and IL-18 KO mice were divided into four groups, including imiquimod (IMQ)-treated IL-18 KO group (w = 11) and WT group (n = 13) as well as their respectively gene-matched control mice (receiving vaseline;w = 12). PASI scores were used to evaluate psoriatic lesions in IMQ-treated mice. Pathological features and dermal cellular infiltration were investigated by hematoxylin and eosin staining. The levels of psoriasis-related cytokines including IL-23, IL-17, IL-12, IL-10, IFNγ, IL-15, IL-27, and IL-4 were tested by real-time polymerase chain reaction (PCR). The protein level of IL-1β, IL-27, CXCL1, and Ly6g were investigated by immunohistochemistry (IHC). Results: Acanthosis (98.46±14.12 vs. 222.68土71.10μm, P<0.01) and dermal cell infiltration (572.25±47.45 vs. 762.47土 59.59cells/field, P< 0.01) were significantly milder in IMQ-induced IL-18 KO mice compared with that in WT mice. IMQ-induced IL-18 KO mice manifested larger areas of Munro microabscesses (11,467.83±5112.09 vs. 4093.19±2591.88 pirn2, P< 0.01) and scales (100,935.24 ±41,167.77 vs. 41,604.41 ± 14,184.10 μm, P<0.01) as compared with WT mice. In skin lesions ot IL-18 KO mice, the expressions of IL-1β, IL-4, and IL-27 were all significantly upregulated but IL-17 was decreased. Histologically, strong positive signals of Ly6g were observed within the epidermis of IL-18 KO mice but expressions of CXCL1 were decreased. Conclusions: IL-18 may exacerbate prominent inflammation and influence pathological features in IMQ-induced mouse model of psoriasis. IL-18 may upregulate pro-inflammatory cytokines and reduce protective cytokines, thus aggravating psoriatic inflammation. In addition, IL-18 may be involved in the formation of Munro microabscesses and scales.

T Helper 1 and T Helper 2 Cytokines Differentially Modulate Expression of Filaggrin and its Processing Proteases in Human Keratinocytes

Background： Atopic dermatitis （AD） is characterized by defective skin barrier and imbalance in T helper 1/T helper 2 （Th 1/Th2） cytokine expression.Filaggrin （FLG） is the key protein to maintaining skin barrier function.Recent studies indicated that Th1/Th2 cytokines influence FLG expression in keratinocytes.However, the role ofThl/Th2 cytokines on FLG processing is not substantially documented.Our aim was to investigate the impact ofThl/Th2 cytokines on FLG processing.Methods： HaCaT cells and normal human keratinocytes were cultured in low and high calcium media and stimulated by either interleukin （IL）-4, 13 or interferon-γ（IFN-γ）.FLG, its major processing proteases and key protease inhibitor lymphoepithelial Kazal-type-related inhibitor （LEKTI） were measured by both real-time quantitative polymerase chain reaction and Western blotting.Their expression was also evaluated in acute and chronic AD lesions by immunohistochemistry.Results： IL-4/13 significantly reduced, while IFN-γsignificantly up-regulated FLG expression.IL-4/13 significantly increased, whereas IFN-γsignificantly decreased the expression ofkallikreins 5 and 7, matriptase and channel-activating serine protease 1.On the contrary, IL-4/13 significantly decreased, while IFN-γincreased the expression of LEKTI and caspase-14.Similar trends were observed in AD lesions.Conclusions： Our results suggested that Th1/Th2 cytokines differentially regulated the expression of major FLG processing enzymes.The imbalance between Th1 and Th2 polarized immune response seems to extend to FLG homeostasis, through the network of FLG processing enzymes.

High Staphylococcus epidermidis Colonization and Impaired Permeability Barrier in Facial Seborrheic Dermatitis

Background：Seborrheic dermatitis （SD） is a common inflammatory skin condition.The etiology is unclear,although overgrowth of Malassezia on the skin has been suggested to cause SD.This study investigated whether colonization with Staphylococcus plays a role in facial SD,which was not well addressed previously.Methods：The study was conducted from September 1,2011 to February 20,2012 in the First Hospital of China Medical University.In the first phase,the study evaluated the level of transepidennal water loss （TEWL） and the number of colony-forming units （CFU） of Staphylococcus in defined skin areas of SD patients who were human immunodeficiency virus （HIV） seropositive （HIV [＋] SD [＋] group,n =13）,classical SD （HIV [-] SD [＋] group,n =24） patients,H IV seropositive-non-SD （HIV [＋] SD [-] group,n =16） patients,and healthy volunteers （HIV [-] SD [-] group,n 16）.In the second phase,we enrolled another cohort of HIV （-） SD （＋） patients who applied topical fusidic acid （n =15）,tacrolimus （n =16）,or moisturizer （n =12）.Changes in the Seborrheic Dermatitis Area Severity Index （SDASI）,TEWL,and Staphylococcus density were evaluated 2 weeks later.Comparisons of each index were performed using analysis of variance （ANOVA） and least significant difference method.Results：The level of TEWL was greater through lesional sites in the HIV （＋） SD （＋） group than that in HIV （＋） SD （-） and HIV （） SD （-） groups （95% confidence interval [CI]：18.873-47.071,P 〈 0.001 and 95% CI：28.755-55.936,P 〈 0.001,respectively）.The number of CFU of Staphylococcus was greater in the HIV （＋） SD （＋） group than that in HIV （＋） SD （-） and HIV （-） SD （-） groups （95% CI：37.487-142.744,P =0.001 and 95% CI：54.936-156.400,P 〈 0.001,respectively）.TEWL was significantly more improved in patients treated with tacrolimus and fusidic acid than that in those treated with moisturizers （95% CI：7.560 38.987,P =0.004 and 95% CI：4.659-37.619,P =0.01 1,respectively）.Topical tacrolimus and fusidic acid were significantly associated with decreased SDASI as compared with moisturizer （95% CI：0.03-0.432,P =0.025 and 95% CI：0.033 0.44,P =0.024,respectively）.Conclusions：High colonization with Staphylococcus epidermidis,along with impaired skin permeability barrier function,contributes to the occurrence of SD.

Malassezia furfur promoting growth of Staphylococcus epidermidis by increasing pH when cultured in a lipid-free environment

To the Editor:Malassezia and Staphylococcus epidermidis(S.epidermidis)are commensal organisms found on human skin,and both display an imbalance of colonization in individuals with seborrheic dermatitis/dandruff(SD/D)[1-3].Malassezia have been implicated in SD/D pathogenesis,as evidenced by an imbalance in the numbers of Malassezia that colonize the skin lesions of people with SD/D,and the fact that antifungal treatment exerts a modifying effect.[4].

Proteomic Analysis of the Serum of Patients with Stable Vitiligo and Progressive Vitiligo

To the Editor： Vitiligo is not a fatal disease; however, it can have strong social and psychological impacts on the patients. Genetic susceptibility, autoimmunity, neural dysregulation, melanin self-destruction, and oxidative stress may be involved in the pathogenesis of vitiligo.

DnaJA4 is involved in responses to hyperthermia by regulating the expression of F-actin in HaCaT cells

Background:Hyperthermia in combination with DnaJA4-knockout(KO)obviously affects the anti-viral immunity of HaCaT cells.The mechanisms of this process are not yet fully explored.However,it is known that DnaJA4 interacts with actin cytoskeleton after hyperthermia.Our aim was to investigate the effects of DnaJA4 on F-actin in HaCaT cells following hyperthermia.Methods:Wild-type(WT)and DnaJA4-KO HaCaT cells were isolated at either 37°C(unheated)or 44°C(hyperthermia)for 30 min followed by testing under conditions of 37°C and assessing at 6,12,and 24 h after hyperthermia.The cytoskeleton was observed with immunofluorescence.Flow cytometry and Western blotting were used to detect the expression of F-actin and relevant pathway protein.Results:DnaJA4-KO and hyperthermia changed the cytoskeleton morphology of HaCaT cells.F-actin expression levels were elevated in DnaJA4-KO cells compared with WT cells(6364.33±989.10 vs.4272.67±918.50,P<0.05).In response to hyperthermia,F-actin expression levels of both WT and DnaJA4-KO cells showed a tendency to decrease followed by an obvious recovery after hyperthermia(WT cells:unheated vs.6 h after hyperthermia or 24 h after hyperthermia:0.34±0.02 vs.0.24±0.01,0.31±0.01,P<0.001,P<0.05;DnaJA4-KO cells:unheated vs.6 h after hyperthermia or 24 h after hyperthermia:0.44±0.01 vs.0.30±0.01,0.51±0.02,P<0.001,P<0.01).WT cells restored to baseline levels observed in the unheated condition,while DnaJA4-KO cells exceeded baseline levels in the recovery.As the upstream factors of F-actin,a similar profile in rho-associated serine/threonine kinase 1(ROCK 1)and RhoA expressions was observed after hyperthermia.While E-cadherin expression was decreased in response to hyperthermia,it was increased in DnaJA4-KO cells compared with WT cells.Conclusions:Hyperthermia affects the expression levels of F-actin in HaCaT cells.DnaJA4 knockout increases the expression of F-actin in HaCaT cells after hyperthermia.DnaJA4 regulates the expressions of F-actin and the related pathway proteins in response to hyperthermia in HaCaT cells.

A 10-year-old Girl with Metastatic Unclassified Sarcoma with Epithelioid Features

To the Editor： A 10-year-old girl presented with an asymptomatic ulcerated tumor on her right forearm for 18 months [Figure 1a]. The minor initially was inconspicuous and then gradually enlarged with painless ulceration in its center [Figure 1b]. Three months later, she developed three nontender new nodules on her right arm, proximal to the initial ulcerated nodule. Her general condition was good. Skin examination revealed a 4 cm × 4 cm tumor, with central ulceration and raised margins. Three diameters in 3-5 cm,