BACKGROUND:Given the proinflammatory cascade elicited by tumor necrosis factor-α (TNF-α) in psoriasis, multiple TNF-α-targeted biologics have been developed for psoriasis treatment. Although systemic macromolecular biologics are widely used, a crucial therapeutic gap remains for mild-to-moderate psoriasis, underscoring an unmet need for more effective topical drugs suppressing TNF-induced inflammatory signaling. OBJECTIVE:To identify a novel potent natural small-molecule drug suppressing TNF-induced inflammatory signaling and elucidate its therapeutic mechanism in psoriasis. METHODS:First, candidate small-molecule drugs were screened out through a high-throughput screening platform. Next, the therapeutic effect of Isolinderalactone was evaluated through topical application in the imiquimod (IMQ)-induced psoriasis-like mouse model. Subsequently, RNA sequencing (RNA-seq) analysis of TNF-α-stimulated HaCaT cells and epidermis of IMQ-treated mice identified key transcriptomic alterations induced by Isolinderalactone treatment. Finally, anti-psoriasis effects and underlying mechanisms of Isolinderalactone were verified in both in vivo and in vitro experiments. RESULTS:Isolinderalactone was identified as a potent drug suppressing TNF-related signaling with low cytotoxicity. Topical application of Isolinderalactone significantly alleviated IMQ-induced psoriasis-like dermatitis. Conjoint analysis of RNA-seq for TNF-α-stimulated HaCaT cells and epidermis from lesions of IMQ-treated mice revealed Isolinderalactone downregulated the expression of TNF-α, interleukin-17 (IL-17) and S100-related inflammatory factors in epidermal keratinocytes. Mechanistically, Isolinderalactone significantly inhibited the TNF-α/STAT3 inflammatory pathways in epidermal keratinocytes and exerted an anti-inflammatory effect. CONCLUSION:Isolinderalactone exhibits anti-inflammatory activity through multiple mechanisms, highlighting the potential of topical Isolinderalactone therapy for mild-to-moderate psoriasis.

We report on two patients with a c.844C>T variant in TINF2 demonstrating distinct phenotypes of classic dyskeratosis congenita (DC) and a novel linear hypermelanosis in narrow bands similar to mosaic DC. The first patient presented with classic features of DC linked to a germline mutation, while the second exhibited extensive linear hypermelanosis in narrow bands similar to the pigmentary lesions attributed to somatic mosaicism. These findings underscore the critical role of genetic analysis, particularly in affected tissues, for understanding mosaic dermatoses. Our report expands the phenotypic spectrum of DC and sheds light on the molecular mechanisms underlying mosaic congenital dyschromatosis.

BACKGROUND:Porokeratosis ptychotropica (PP) is a severe inherited skin disorder resistant to traditional treatments. Topical statins are being explored as a potential alternative. OBJECTIVE:To evaluate the efficacy and safety of topical simvastatin and simvastatin/cholesterol in treating PP. METHODS:In this single-blind, split-body, placebo-controlled trial, PP patients were 1:1 randomized to apply either 2% simvastatin or 2% simvastatin/cholesterol cream on 1 randomized side of the buttocks twice daily for 8 weeks. If blinded evaluation determined that the treatment side showed superior efficacy over placebo at week 8, both sides would continue treatment for 40 more weeks. RESULTS:Of 24 participants screened, 18 were enrolled and underwent randomization. One patient withdrew from each group before preliminary assessment. After 8 weeks, 8 of 8 patients in the 2% simvastatin group and 7 of 8 patients in the 2% simvastatin/2% cholesterol group showed better efficacy than placebo. By week 48, severity index and pruritus scores continued to improve, and no difference was seen between the 2 groups. No treatment-related adverse events were observed. LIMITATIONS:This trial had a relatively small sample size. CONCLUSION:Both topical simvastatin and simvastatin/cholesterol were well tolerated and effective in relieving the erythema, thickness, scaling, and pruritus of PP with relatively long-term use.

BACKGROUND:Nagashima-type palmoplantar keratoderma (NPPK), caused by biallelic SERPINB7 loss-of-function variants, lacks effective treatments. Intriguingly, monoallelic exonic variants are observed in some patients with NPPK, suggesting additional genetic variants. OBJECTIVE:To characterize a deep intronic SERPINB7 variant's pathogenicity, elucidate its splicing dysregulation, and evaluate antisense oligonucleotide (ASO) therapy. METHODS:A customized next-generation sequencing panel was applied to six Chinese NPPK patients. Pathological changes were analyzed by H&E staining and immunofluorescence. RNA extracted from palmar skin was assessed for splicing alterations. Plasmids were generated to evaluate the expression and function of mutant SERPINB7 protein. Haplotype analysis was conducted to confirm the founder effect. RNA pull-down assays and mass spectrometry were used to identify the key splicing factor. Minigene constructs were developed to characterize the splicing process in vitro. Finally, an ASO was designed to target this variant. RESULTS:A deep intronic SERPINB7 variant was identified in six NPPK patients, leading to pseudo-exon inclusion and the production of a truncated, dysfunctional SERPINB7 protein. Haplotype analysis confirmed it as a Chinese founder variant. RNA pull-down assays revealed excessive SRSF9 binding to the abnormal transcript. In vitro, the ASO successfully corrected the aberrant splicing. CONCLUSION:This study established the pathogenicity of a deep intronic founder variant in SERPINB7 driving NPPK via SRSF9-mediated splicing dysregulation, demonstrating ASO therapeutic potential. Findings provide mechanistic insights and a targeted approach for precision therapy development for NPPK.

Inherited ichthyosis comprises a spectrum of genetic disorders related to over 50 pathogenic genes. However, there are limited data summarizing the clinical and molecular characteristics of Chinese patients. To broaden the knowledge of clinical and genetic characteristics of inherited ichthyosis and to optimize disease diagnosis and therapies, cases diagnosed with inherited ichthyosis in 1 tertiary centre from 2019 to 2023 were collected, excluding ichthyosis vulgaris and X-linked recessive ichthyosis, genomic sequencing was then performed, and clinical details of the patients were assessed. A total of 35 patients from Jiangsu and Anhui provinces of China were enrolled, 31 of whom were diagnosed with non-syndromic ichthyosis. Within this group, there were cases of autosomal recessive congenital ichthyosis (18/31), epidermolytic ichthyosis (9/31), and superficial epidermolytic ichthyosis (4/31). Additionally, 4 patients were diagnosed with syndromic ichthyosis, comprising 1 case of Chanarin-Dorfman syndrome and 3 cases of Netherton syndrome. The genetic analysis revealed a total of 47 variants across 13 genes, of which 19 were identified as novel variants. This study describes the clinical spectrum of rare inherited ichthyosis in the Jiangsu-Anhui region of China and further expands the genetic characteristics of the disease.

Background: Conventional systemic corticosteroid therapy for bullous pemphigoid (BP) has been challenged due to severe adverse events. Dupilumab has emerged as an alternative therapeutical option of BP patients. Objectives: To evaluate the efficacy of dupilumab monotherapy and the combination with medium/low-dose corticosteroids for BP treatment. Methods: Thirteen, twenty-four and thirty-two BP patients treated with Dupilumab monotherapy (Dupi group), dupilumab combined with corticosteroids (Dupi + CS group), and corticosteroid monotherapy (CS group), respectively, were retrospectively analyzed for various clinical and laboratory parameters. Results: In the Dupi group, the total Bullous Pemphigoid Disease Area Index (BPDAI) Total, Erosion/Blister, Urticaria/Erythema and Itching NRS scores were all reduced significantly after 2--4 weeks of treatment, but the BPDAI Mucosal Score was not changed significantly at the end of the overextended time of treatment. All the above clinical parameters and many laboratory parameters (including the serum anti-BP180 autoantibodies [IgG] level, blood eosinophil count, and percentage) were significantly reduced in both Dupi + CS and CS groups after treatment, but no statistical differences were found in the reduction rates of these parameters between the two groups. However, the Dupi + CS group had less baseline dose and cumulative dosage of prednisone at the time of disease control, and fewer adverse effects were reported than the CS group. Study limitations: The retrospective design and small clinical sample size of the Dupi group. Conclusions: For BP patients, dupilumab monotherapy based on the treatment of atopic dermatitis can significantly improve skin lesions and pruritus symptoms but may be ineffective for oral mucosal lesions. The combination of dupilumab and medium/low-dose corticosteroids can achieve the same effect of corticosteroid therapy with superior safety. (c) 2024 Sociedade Brasileira de Dermatologia. Published by Elsevier Espana, S.L.U. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).

Acne is a common chronic inflammatory disease of the pilosebaceous unit. Transient receptor potential vanilloid 3 (TRPV3) is an ion channel that is involved in inflammatory dermatosis development. However, the involvement of TRPV3 in acne-related inflammation remains unclear. Here, we used acne-like mice and human sebocytes to examine the role of TRPV3 in the development of acne. We found that TRPV3 expression increased in the skin lesions of Propionibacterium acnes (P. acnes)-injected acne-like mice and the facial sebaceous glands (SGs) of acne patients. TRPV3 promoted inflammatory cytokines and chemokines secretion in human sebocytes and led to neutrophil infiltration surrounding the SGs in acne lesions, further exacerbating sebaceous inflammation and participating in acne development. Mechanistically, TRPV3 enhanced TLR2 level by promoting transcriptional factor p-FOSL1 expression and its binding to the TLR2 promoter, leading to TLR2 upregulation and downstream NF-κB signaling activation. Genetic or pharmacological inhibition of TRPV3 both alleviated acne-like skin inflammation in mice via the TL2-NF-κB axis. Thus, our study revealed the critical role of TRPV3 in sebaceous inflammation and indicated its potential as an acne therapeutic target.

Nagashima-type palmoplantar keratosis, caused by biallelic loss-of-function variants in SERPINB7, is the most prevalent palmoplantar keratoderma in East Asia and manifests as diffuse palmoplantar erythema and hyperkeratosis. We report four different variants of SERPINA12 in six Chinese individuals and identified the recurrent variant c.970_971del as a founder variant among East Asians.

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TRPV3 is a temperature-sensitive calcium-permeable channel. In previous studies, we noticed prominent TUNELpositive keratinocytes in patients with Olmsted syndrome and Trpv3 + /G568V mice, both of which carry gain-of- function variants in the TRPV3 gene. However, it remains unclear how the keratinocytes die and whether this process contributes to more skin disorders. In this study, we showed that gain-of-function variant or pharmacological activation of TRPV3 resulted in poly(ADP-ribose) polymerase 1 (PARP1)/AIFM1/macrophage migration inhibitory factor axis- mediated parthanatos, which is an underestimated form of cell death in skin diseases. Chelating calcium, scavenging ROS, or inhibiting nitric oxide synthase effectively rescued the parthanatos, indicating that TRPV3 regulates parthanatos through calcium-mediated oxidative stress. Furthermore, inhibiting PARP1 downregulated TSLP and IL33 induced by TRPV3 activation in HaCaT cells, reduced immune cell infiltration, and ameliorated epidermal thickening in Trpv3 + /G568V mice. Marked parthanatos was also detected in the skin of MC903-treated mice and patients with atopic dermatitis, whereas inhibiting PARP1 largely alleviated the MC903-induced dermatitis. In addition, stimulating parthanatos in mouse skin with methylnitronitrosoguanidine recapitulated many features of atopic dermatitis. These data demonstrate that the TRPV3-regulated parthanatosassociated PARP1/AIFM1/macrophage migration inhibitory factor axis is a critical contributor to the pathogenesis of Olmsted syndrome and atopic dermatitis, suggesting that modulating the PARP1/AIFM1/macrophage migration inhibitory factor axis is a promising therapy for these conditions.

Nagashima-type palmoplantar keratoderma (NPPK) is an autosomal recessive genodermatosis caused by loss-of-function variants in SERPINB7 and is the most prevalent form of inherited palmoplantar keratodermas among Asians. However, there is currently no effective therapy for NPPK because its pathogenesis remains unclear. In this study, Serpinb7-/- mice were generated and spontaneously developed a disrupted skin barrier, which was further exacerbated by acetone-ether-water treatment. The skin of these Serpinb7-/- mice showed weakened cytoskeletal proteins. Additionally, SERPINB7 deficiency consistently led to decreased epidermal differentiation in a three-dimensional human epidermal model. We also demonstrated that SERPINB7 was an inhibitory serpin that mainly inhibited the protease legumain. SERPINB7 bound directly with legumain and inhibited legumain activity both in vitro and in vivo. Furthermore, we found that SERPINB7 inhibited legumain in a ‘protease-substrate’ manner and identified the cleavage sites of SERPINB7 as Asn71 and Asn343. Overall, we found that SERPINB7 showed the nature of a cysteine protease inhibitor, and identified legumain as a key target protease of SERPINB7. Loss of SERPINB7 function led to overactivation of legumain, which might disrupt cytoskeletal proteins, contributing to the impaired skin barrier in NPPK. These findings may lead to the development of therapeutic strategies for NPPK.

Start codon variants in ubiquitin ligase KLHL24 lead to a gain-of-function mutant KLHL24-ΔN28, which mediates the excessive degradation of keratin 15, desmin, and keratin 14, resulting in alopecia, cardiopathy, and epidermolysis bullosa syndrome. Patients with alopecia, cardiopathy, and epidermolysis bullosa syndrome normally present atrophic scars after wounds heal, which is rare in KRT14-related epidermolysis bullosa. The mechanisms underlying the formation of atrophic scars in epidermolysis bullosa of patients with alopecia, cardiopathy, and epidermolysis bullosa syndrome remain unclear. This study showed that KLHL24-ΔN28 impaired skin wound healing by excessively degrading vimentin. Heterozygous Klhl24c.3G>T knock-in mice displayed delayed wound healing and decreased wound collagen deposition. We identified vimentin as an unreported substrate of KLHL24. KLHL24-ΔN28 mediated the excessive degradation of vimentin, which failed to maintain efficient fibroblast proliferation and activation during wound healing. Furthermore, by mediating vimentin degradation, KLHL24 can hinder myofibroblast activation, which attenuated bleomycin-induced skin fibrosis. These findings showed the function of KLHL24 in regulating tissue remodeling, atrophic scarring, and fibrosis.

Bullous pemphigoid (BP) is a complex inflammatory process with elevated levels of autoantibodies, eosinophils, neutrophils, and various cytokines. Hematological inflammatory biomarkers can reflect inflammatory state in various diseases. Up to now, the correlations of hematological inflammatory biomarkers and disease activity of BP remain unknown. The purpose of this study was to clarify the associations between hematological inflammatory biomarkers and disease activity of BP. The levels of neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR), platelet-to-neutrophil ratio (PNR) and mean platelet volume (MPV) of 36 untreated BP patients and 45 age and gender matched healthy controls were detected by routine blood tests. The correlations between hematological inflammatory markers and clinical characteristics of BP were statistically analyzed. The Bullous Pemphigoid Disease Area Index (BPDAI) was used to measure disease activity of BP. The mean levels of NLR, PLR, PNR and MPV in 36 untreated BP patients were 3.9, 157.9, 45.7 and 9.4 fl, respectively. Increased NLR (p < 0.001), PLR (p < 0.01), and MPV (p < 0.001) but decreased PNR (p < 0.001) were observed in BP patients when compared with healthy controls. In BP patients, the levels of NLR were positively correlated to BPDAI Erosion/Blister Scores (p < 0.01); and the levels of NLR and PLR were both positively correlated to BPDAI without Damage Score (both p < 0.05) and BPDAI Total Score (both p < 0.05). No correlation was found in other statistical analyses between hematological inflammatory markers and clinical characteristics in BP patients involved in the present study. Therefore, NLR and PLR are positively correlated with disease activity of BP.

The correlation between IgE anti-BP180 NC16A autoantibody and disease activity of bullous pemphigoid (BP) remains disputable. To determine the levels of IgE anti-BP180 NC16A autoantibody and its clinical significance in untreated BP patients. IgG and IgE anti-BP180 NC16A autoantibody in serum and blister fluid samples of 34 untreated BP patients was detected by enzyme-linked immunosorbent assay (ELISA), and correlation with clinical and pathological features of BP were statistically analysed. The Bullous Pemphigoid Disease Area Index (BPDAI) was used to measure disease activity of BP. The mean baseline level of IgG anti-BP180 NC16A autoantibody in serum and blister fluid samples of untreated BP patients was 75.3 U/mL and 1.54 U/mL, respectively (A450, cutoff: 0.126). IgE anti-BP180 NC16A autoantibody was positive in 21.9% serum and 14.7% blister fluid samples of untreated BP patients. IgE anti-BP180 NC16A autoantibody levels in serum samples positively correlated with those from blister fluid samples (r = 0.983, p < 0.05). However, IgE anti-BP180 NC16A autoantibody level in both serum and blister fluid samples of untreated BP patients did not correlate with IgG anti-BP180 NC16A autoantibody, age, extent of elevated peripheral blood eosinophils, BPDAI erosion/blister score, BPDAI urticaria/erythema score, BPDAI pruritus score, BPDAI without damage score, or BPDAI total score (all p > 0.05). No significant correlation was identified between disease activity and positive or negative anti-BP180 NC16A IgE autoantibody. Conclusion: IgE anti-BP180 NC16A autoantibody in both serum and blister fluid samples does not appear to correlate with disease activity of BP.

1 Schulz R, Beach SR. Caregiving as a risk factor for mortality: the Caregiver Health Effects Study. JAMA 1999; 282:2215–19. 2 Vitaliano PP, Zhang J, Scanlan JM. Is caregiving hazardous to one’s physical health? A meta-analysis. Psychol Bull 2003; 129:946–72. 3 Bridgman AC, Block JK, Drucker AM. The multidimensional burden of atopic dermatitis: an update. Ann Allergy Asthma Immunol 2018; 120:603–6. 4 Drucker AM, Wang AR, Li WQ et al. The burden of atopic dermatitis: summary of a report for the National Eczema Association. J Invest Dermatol 2017; 137:26–30. 5 Kim RW, Barta K, Begolka WS et al. Qualitative analysis of the impact of atopic dermatitis on caregivers. Br J Dermatol 2022; doi: https://doi. org/10.1111/bjd.21828. 6 Elsawi R, Dainty K, Begolka W et al. The multidimensional burden of atopic dermatitis among adults: results from a large national survey. JAMA Dermatol 2022; 158:887–92. 7 Xu X, van Galen LS, Koh MJA et al. Factors influencing quality of life in children with atopic dermatitis and their caregivers: a crosssectional study. Sci Rep 2019; 9:15990. 8 Kudra A, Lees C, Morrell-Scott N. Measuring carer burden in informal carers of patients with long-term conditions. Br J Community Nurs 2017; 22:230–6.

To the Editor: Laboratory diagnostic methods of bullous pemphigoid (BP) include direct immunofluorescence (DIF) microscopy, BP180 NC16A enzyme-linked immunosorbent assays (ELISA), and indirect immunofluorescence using salt-split skin (ssIIF) in our daily clinical practice. Immunofluorescence (IF) studies are important parts of the laboratory workup for BP. DIF is regarded as the gold standard for BP diagnosis; however, it is not specific and can also be found in other acquired autoimmune subepidermal blistering dermatosis. ssIIF was found to have a specificity of 100% for BP diagnosis and in some guidelines, it is recommended to perform ssIIF in every patient with clinically suspected BP.[1] To date, it is still not defined whether ssIIF can be used as a routine diagnosis method of BP. The BP180 NC16A ELISA is a practical and reliable diagnostic test for BP; however, the false-positive results of BP180 NCA6A ELISA were recently reported in a wide range of dermatoses, which might cause its uncertainty in the diagnosis of BP. Thus, we conducted a single center retrospective study to compare diagnostic values of these tests for BP from Hospital for Skin Diseases, Chinese Academy of Medical Sciences between January 2014 and January 2019. It was approved by the local medical ethical committee (No. 2017-KY-022) and consent was obtained from patients prior to serum collection. In this study, we analyzed the data of 569 patients who were tested by ssIIF, DIF, BP180 NC16A ELISA and histopathology at the initial disease stage prior to systemic therapy, and were followed for at least 1 year, and BP diagnosis was made on the basis of the following criteria: (1) tense bullae on body without scaring, and Nikolsky sign is negative; (2) histopathological finding shows subepidermal blister formation; (3) DIF shows linear deposits of IgG and/or C3 along the basement membrane zone (BMZ); (4) ssIIF shows linear IgG and/or C3 reaction with the epidermal side of BMZ in the salt-split skin; (5) index value of BP180 ELISA is more than 9 U/mL. Diagnosis of BP requires at least two columns, (4) and (5), or requires three columns, (1), (2), and (3) if it meets one column of (4) and (5). Based on these diagnosis criteria, 164 of 569 suspected BP patients were diagnosed with BP, and the remaining 405 patients were used as non-BP controls. Study flowchart and characteristics of patients are described in Supplementary Figure 1, https://links.lww.com/CM9/B92 and Supplementary Table 1, https://links.lww.com/CM9/B92. Sensitivity and specificity were calculated for assessing diagnostic values of DIF, ssIIF and BP180 NC16A ELISA, and Kappa test was used to evaluate the consistency between these tests. Measurement data were shown as mean ± standard deviation, and were compared by Mann-Whitney U test or paired t-test. Numeration data with rate or composition ratio were analyzed by McNemar test, χ2 test or Fisher's exact test. Statistical analyses were done using SPSS statistics 22 (IBM Corp, Armonk, NY, USA). All tests were two-sided, and P < 0.05 was considered as statistically significant. For diagnosis of BP, the sensitivity of DIF, ssIIF, and BP180 ELISA were 78.05%, 97.56%, 93.90%; and their specificity were 93.58%, 99.75%, and 95.55%, respectively [Supplementary Table 2, https://links.lww.com/CM9/B92]. In previous studies, the specificity of DIF and ssIIF was 98% to 99%, 97% to 100%, and the sensitivity was 88% to 91%, 73% to 97%, respectively.[2,3] The specificity of DIF was similar to the findings of the studies mentioned above, and further proved that DIF was not specific enough for BP diagnosis. Some researchers found routine serration pattern analysis can help to improve the specificity of DIF in BP. The sensitivity of DIF in our data is lower than the studies mentioned above, and the reason may be due to availability of serologic test data, or ethnic differences, which was supported by another domestic article, and their positive rate of DIF is 82%.[4] Some researchers found IgG4 testing may be helpful in definitively diagnosing BP in patients with negative or equivocal linear IgG deposition on DIF.[5] In this study, the high specificity of ssIIF was consistent with the reports mentioned above, and the sensitivity was similar to the recent study from 13 international centers,[3] but higher than the previous report.[2] The relative higher sensitivity may be caused by transporting and tackling skin tissue timely which can avoid protein denaturation and degradation. Given all the data of ssIIF on BP diagnosis, we considered ssIIF as a valuable method for BP diagnosis. In the present study, we found that BP180 ELISA results were negative in 6.1% BP patients, which might be caused by lower titers of anti-BP180 NC16A antibodies or existence of autoantibodies recognizing BP180 on non-NC16A domain and/or BP230 in BP sera. The results indicated that it needed to be combined with IF to avoid a misdiagnosis. Next, the sensitivity and specificity of combined tests were analyzed. The sensitivity of ssIIF/BP180 ELISA (91.66%) is statistically significantly higher than those of ssIIF/DIF (75.61%), DIF/BP180 ELISA (71.95%), and the specificities of three combined tests are all 100% [Supplementary Table 2, https://links.lww.com/CM9/B92]. These results indicated that combined tests could obtain perfect specificity, although the sensitivity is decreased a little bit, and combination of ssIIF/BP180 ELISA has the highest sensitivity in BP diagnosis. In addition, Kappa coefficients between ssIIF/BP180 ELISA, ssIIF/DIF, and DIF/BP180 ELISA were 0.86, 0.706, 0.622, respectively (all P < 0.001). And BP180 ELISA was positively correlated with ssIIF in BP patients [Supplementary Table 3, https://links.lww.com/CM9/B92]. The limitation of this study is the absence of diagnostic criteria as a reference standard for the diagnosis of BP. Patients were diagnosis as BP by at least both positive ssIIF and positive BP180 NC16A ELISA, or a combination of clinical bulla, subepidermal bulla on routine histopathology and positive DIF with either positive ssIIF or positive BP180 NC16A ELISA which may bias the sensitivity and specificity of these three diagnostic methods. In summary, ssIIF could be used as a routine laboratory diagnosis method for BP, and the combination of ssIIF and BP180 ELISA is better for diagnosis of BP. Funding This work was supported by grants from Chinese Academy of Medical Sciences (CAMS) Innovation Fund for Medical Sciences (Nos. CIFMS-2017-I2M-1-017, CIFMS-2021-I2M-1-059), and Scientific Research Project of Jiangsu Provincial Health Commission (No. ZD2021035). 4njkmnjm Conflicts of interest None.

Background: TNF-alpha elicits a cascade amplification effect in psoriasis. Macromolecule drugs targeting TNF-alpha are widely used for the clinical treatment of psoriasis. However, there are currently no effective small -molecule inhibitors that can be used in the clinic. Objective: Novel TNF-alpha inhibitor was identified via high-throughput screening (HTS) and its anti-in-flammatory activity was evaluated.Methods: Two cell death models were established to identify inhibitors of TNF-alpha through HTS from a library of 3256 compounds. The effect of the inhibitor of TNF-alpha was tested by HaCaT cells in vitro and IMQ-induced psoriasis-like mouse model in vivo.Results: Tiamulin fumarate (TF) was identified as an effective inhibitor of TNF-alpha. TF significantly blocked the NF-Kappa B and MAPK signaling pathways in TNF-alpha-stimulated HaCaT cells. Additionally, systemic and topical administration of TF improved IMQ-induced psoriasis-like dermatitis in the mouse model.Conclusion: Our study established a HTS method to identify TF as an inhibitor of TNF-alpha. The protective roles of TF in psoriasis-related inflammation reveal the potential therapeutic value of TF for psoriasis.(c) 2022 Japanese Society for Investigative Dermatology. Published by Elsevier B.V. All rights reserved.

Objective:To optimize indirect immunofluorescence on salt-split skin (IIF-SSS), and to evaluate its performance in detection of bullous pemphigoid (BP) antibodies.Methods:Normal human foreskin and non-foreskin skin tissues were used to prepare salt-split substrates under 3 different experimental conditions: traditional group rotated at 4 ℃ for 48 - 72 hours, low-temperature immersion group soaked at 4 ℃ for 48 - 72 hours, room-temperature immersion group soaked at 25 ℃ (range: 23 - 27 ℃) for 24 hours. Serum samples were obtained from 20 patients with bullous pemphigoid (BP) in Hospital of Dermatology, Chinese Academy of Medical Sciences between August 2019 and August 2020, and subjected to IIF on the intact skin or salt-split substrates by using a multiple dilution method. Paired-sample t test was used for comparisons of means between two paired samples. Results:No dermal-epidermal separation was observed in the substrates prepared in the low-temperature immersion group at 48 - 72 hours, while dermal-epidermal separation occurred in the lower lamina lucida of the foreskin and non-foreskin substrates in the room-temperature immersion group and the traditional group. For the 20 patients with BP, the reciprocal end-point titers ( M[ Q1, Q3]) detected with the salt-split non-foreskin skin and salt-split foreskin in the room-temperature immersion group, and with the salt-split non-foreskin skin in the traditional group were 5 120 (2 560, 17 920), 1 280 (640, 2 560), 1 280 (640, 2 560), respectively. Moreover, 19 (95%) patients with BP showed that the reciprocal end-point titers detected with the substrates in the room-temperature immersion group were 1 - 5 times those in the traditional group ( t = 8.04, P<0.001), suggesting that the performance of salt-split skin in the room-temperature immersion group was superior to that in the traditional group in the detection of BP antibodies; however, there was no significant difference in the reciprocal end-point titers of BP antibodies between the salt-split foreskin in the room-temperature immersion group and salt-split non-foreskin skin in the traditional group ( t<0.001, P>0.05). The reciprocal end-point titers in 20 BP sera detected by conventional IIF on the intact non-foreskin skin and foreskin were 320 (160, 640) and 480 (160, 1 120), respectively; the reciprocal end-point titers detected by IIF on the salt-split foreskin and non-foreskin skin in the room-temperature immersion group, as well as on the salt-split non-foreskin skin in the traditional group, were all consistent with or 1 - 7 times higher than those detected by conventional IIF ( t = 6.47, 14.83, 5.26, respectively, all P<0.001) . Conclusion:The soaking method at room temperature 25 ℃ (23 - 27 ℃) for preparing salt-split substrates has advantages of short duration and simple procedure, and the sensitivity of IIF-SSS using the substrates prepared by this method is equal or superior to the traditional salt-split method for detecting BP antibodies.

Objective:To evaluate the value of indirect immunofluorescence on salt-split skin (IIF-SSS) in the diagnosis of bullous pemphigoid (BP) .Methods:A single-center clinical retrospective study was conducted. Totally, 163 patients with newly diagnosed BP were collected from Hospital of Dermatology, Chinese Academy of Medical Sciences from January 2013 to January 2019, so were 404 controls, including 161 with pemphigus, 67 with eczema, 26 with drug eruption, 23 with erythema multiforme, 18 with prurigo nodularis, etc. Blood samples were collected before the treatment, and IIF-SSS, BP180 NC16A enzyme-linked immunosorbent assay (ELISA) and direct immunofluorescence (DIF) assay were performed to evaluate the value of IIF-SSS in the diagnosis of BP. Measurement data were compared by using t test and Mann-Whitney test, and enumeration data were compared by using chi-square test and Fisher′s exact test or McNemar test. Results:The number of cases positive for IIF-SSS, BP180 NC16A ELISA and DIF assay was 160, 153 and 127 respectively in the BP group, and 0, 18 and 26 respectively in the control group. The sensitivities of IIF-SSS, BP180 NC16A ELISA and DIF assay for the diagnosis of BP were 98.15%, 93.86% and 77.91% respectively, and their specificities were 100%, 95.54% and 93.56% respectively. There was strong consistency in the diagnosis of BP between IIF-SSS and DIF (Kappa coefficient= 0.767, P < 0.001) . Conclusion:IIF-SSS has relatively high sensitivity and specificity for the diagnosis of BP, and can serve as a routine method for diagnosing BP.

Objective:To investigate the correlation of related antibody titers in serum of patients with pemphigus vulgaris (PV) with disease severity and activity.Methods:A total of 24 patients with active PV were collected, who firstly visited Hospital for Skin Diseases, Chinese Academy of Medical Sciences from 2012 to 2015. Pemphigus disease area index (PDAI) was evaluated in the patients with PV at active and stable stages, and serum samples were collected. Enzyme-linked immunosorbent assay (ELISA) was conducted to determine titers of pathogenic anti-desmoglein (Dsg) conformational epitope antibodies, total anti-Dsg antibodies and anti-acetylcholine receptor (AChR) antibody in serum samples. Measurement data were compared by using t test, enumeration data were compared by using Fisher′s exact test, and correlations were analyzed by using Pearson correlation analysis. Results:Among the patients with active PV, there was no significant difference between the anti-Dsg1 antibody titers (611.4 ± 136.8) and anti-Dsg1 conformational epitope antibody titers (585.5 ± 134.7, t = 0.13, P = 0.89) , but the anti-Dsg3 antibody titers (708.6 ± 130.7) were significantly higher than the anti-Dsg3 conformational epitope antibody titers (297.2 ± 54.4, t = 2.90, P < 0.01) . In addition, both the anti-Dsg1 antibody titers and anti-Dsg1 conformational epitope antibody titers were positively correlated with PDAI scores in the patients with active PV (both r = 0.54, P < 0.01) ; PDAI scores were not correlated with the anti-Dsg3 antibody titers ( r = 0.11, P = 0.62) , but positively correlated with the anti-Dsg3 conformational epitope antibody titers ( r = 0.53, P < 0.01) . Among the 20 patients with stable PV, the serum titers of anti-Dsg1 antibodies and anti-Dsg1 conformational epitope antibodies significantly decreased compared with those at their first visit; anti-Dsg3 antibody titers significantly decreased in only 7 patients, and 13 patients still had high titers of anti-Dsg3 antibodies, including 6 with declined anti-Dsg3 conformational epitope antibody titers, and 5 converted from anti-AChR antibody-positive to anti-AChR antibody-negative. Conclusions:Both the anti-Dsg1 antibody and anti-Dsg1 conformational epitope antibody titers can reflect the disease activity of PV. The disease activity was not consistent with anti-Dsg3 antibody titers in some patients, and the anti-Dsg3 conformational epitope antibody or anti-AChR antibody may facilitate evaluating the disease activity of PV.