Reflectance confocal microscopy provides real-time images of vulvar lichen sclerosus at cellular resolution.

Abstract

Incorrect and delayed diagnosis of vulvar high-grade squamous intraepithelial neoplasia (vHSIL) and lichen sclerosus (LS) increases malignant progression risks and negatively impacts prognosis and quality of life. There is a need to improve diagnosis and monitoring. Reflectance confocal microscopy is a non-invasive imaging tool that visualizes skin structures at cellular resolution. The objectives were to explore feasibility and patient acceptability of vulvar RCM imaging and to identify RCM characteristics that are discriminative for vulvar HSIL and LS. This was a prospective, cross-sectional, observational clinical trial in patients with vHSIL and LS compared to healthy volunteers. RCM images and vulvar tissue samples were obtained. Five (5) patients with vHSIL, 10 patients with LS and 10 healthy volunteers were enrolled. In total, 100 image series of vulvar skin were obtained, including lesional and nonlesional sites. The RCM technique was considered acceptable for application by patients and healthy controls. Healthy vulvar skin was characterized by a homogenous, normal honeycomb patterned epidermis and a clear epidermal-dermal junctions. Vulvar HSIL and LS displayed an atypical honeycomb pattern of the epidermis and lymphocytic influx with presence of melanophages. Distinct features specifically observed in LS included the presence of hyalinised vessels and sclerotic areas in the dermis. RCM is a non-invasive imaging technique that is feasible and clinically acceptable to apply on vulvar skin, both in patients with premalignant lesions and healthy controls. Recognition and validation of disease-specific characteristics could make reflectance confocal microscopy a clinical tool to non-invasively aid identification of vulvar premalignancies.

The stratum corneum depth profiles of lesional (LPS) and non-lesional psoriatic skin (nLPS) were measured in vivo non-invasively using confocal Raman micro-spectroscopy. The results showed lower lipid concentration, more gauche-conformers, higher hexagonal lateral organisation of intercellular lipids, lower concentration of NMF, higher degree of folded keratin, lower total water and a strong tendency towards less strongly bound and more weakly bound water molecules in LPS. These findings clearly indicate a reduced skin barrier function in LPS compared with nLPS and healthy skin.

Abstract

Psoriasis, one of the most common skin diseases affecting roughly 2%–3% of the world population, is associated with a reduced skin barrier function (SBF) that might play an important role in its pathophysiology. The SBF is provided primarily by the stratum corneum (SC) of the skin. Previous studies have revealed a higher trans-epidermal water loss, lower hydration, abnormal concentration and composition of intercellular lipids, as well as alterations in secondary keratin structure in the psoriatic SC. We compared on molecular level lesional psoriatic skin (LPS) with non-lesional psoriatic skin (nLPS) from 19 patients non-invasively in vivo, using confocal Raman micro-spectroscopy. By analysing the corresponding Raman spectra, we determined SBF-defining parameters of the SC depth-dependently. Our results revealed a lower total lipid concentration, a shift of lamellar lipid organisation towards more gauche-conformers and an increase of the less dense hexagonal lateral packing of the intercellular lipids in LPS. Furthermore, we observed lower natural moisturising factor concentration, lower total water as well as a strong tendency towards less strongly bound and more weakly bound water molecules in LPS. Finally, we detected a less stable secondary keratin structure with increased β-sheets, in contrast to the tertiary structure, showing a higher degree of folded keratin in LPS. These findings clearly suggest structural differences indicating a reduced SBF in LPS, and are discussed in juxtaposition to preceding outcomes for psoriatic and healthy skin. Understanding the alterations of the psoriatic SC provides insights into the exact pathophysiology of psoriasis and paves the way for optimal future treatments.

Abstract

Exposure to the sun affects the skin and may eventually result in UV-induced skin damage. It is generally known that hyaluronan (HA) is one of the main structural and functional components of the skin. However, UV-related changes in the HA metabolism in the skin have not yet been elucidated. Using qRT-PCR, confocal microscopy and LC–MS/MS we compared the naturally sun-exposed (SE), sun-protected, experimentally repeatedly UVA + UVB-exposed and acutely (once) UVA + UVB irradiated skin of Caucasian women. The epidermis was harvested by means of suction blistering 24 h after the acute irradiation. In addition, the epidermis was compared with a UV-irradiated in vitro reconstituted 3D epidermis (EpiDerm) and an in vitro 2D culture of normal human keratinocytes (NHEK). The amount of HA was found to be statistically significantly enhanced in the acutely irradiated epidermis. The acute UV evinced the upregulation of HA synthases (HAS2 and HAS3), hyaluronidases (HYAL2 and HYAL3), Cluster of differentiation 44 (CD44), and Cell Migration Inducing Proteins (CEMIP and CEMIP2), while only certain changes were recapitulated in the 3D epidermis. For the first time, we demonstrated the enhanced gene and protein expression of CEMIP and CEMIP2 following UV irradiation in the human epidermis. The data suggest that the HA metabolism is affected by UV in the irradiated epidermis and that the response can be modulated by the underlying dermis.

Abstract

As near-infrared radiation (NIR), which is a composition of sunlight with an 780–1400 nm wavelength, is associated with skin aging such as wrinkles and slacks, the biological actions of NIR with high dermal penetration remains unclear. In the present study, we found that NIR irradiation (40 J/cm²) at different levels of irradiance (95–190 mW/cm²) using a laboratory device with a xenon flash lamp (780–1700 nm) caused sebaceous gland enlargement concomitantly with skin thickening in the auricle skin of hamsters. The sebaceous gland enlargement resulted from the proliferation of sebocytes due to an increase in the number of proliferating cell nuclear antigen (PCNA)- and lamin B1-positive cells in vivo. In addition, NIR irradiation transcriptionally augmented the production of epidermal growth factor receptor (EGFR) accompanied with an increase in the reactive oxygen species (ROS) level in hamster sebocytes in vitro. Furthermore, the administration of hydrogen peroxide increased the level of EGFR mRNA in the sebocytes. Therefore, these results provide novel evidence that NIR irradiation causes the hyperplasia of sebaceous glands in hamsters by mechanisms in which EGFR production is transcriptionally augmented through ROS-dependent pathways in sebocytes.

Abstract

Mild acidification caused by transcutaneous administration of carbon dioxide (CO₂) has been reported to improve some epidermal skin impairments, such as desquamation and inflammation; however, its effects on dermal tissue remain unclear. Here, we examined the effect and mechanism of mild acidity on extracellular matrix (ECM) protein production in normal human dermal fibroblasts (NHDFs). To achieve this, the skin permeability of CO₂ and its effect on intradermal pH were evaluated by treating reconstructed human skin equivalents (HSEs) with a CO₂-containing formulation. Additionally, NHDFs were cultured in a pH-adjusted medium (pH 6.5). CO₂ successfully permeated HSEs and reduced the intradermal pH. Decreased extracellular pH activated CREB, upregulated TGF-β1 expression, promoted the production of elastic and collagen fibres, and increased hyaluronan concentration in NHDFs. Additionally, the low pH-induced increase in TGF-β1 expression was attenuated via the RNAi-mediated suppression of the expression of CREB1 and proton-sensing G protein-coupled receptors (GPCRs), including GPR4 and GPR65. Moreover, low pH-induced CREB activation was suppressed by the inhibition of the cAMP/PKA and PLC/PKC signalling pathways. Taken together, a CO₂-induced decrease in intradermal pH may promote ECM production in NHDFs via the upregulation of TGF-β1 expression, which was mediated by the activation of the GPCR signalling pathway and CREB, indicating that CO₂ could be used to treat ultraviolet radiation-induced photoaging, intrinsic ageing and ECM deterioration.

Abstract

The dysregulation of branched-chain amino acid (BCAA) metabolism and related enzymes has been greatly implicated in the progression of multiple types of cancer, whereas remains far from understood in melanoma. Here, we explored the role of the BCAA metabolism enzyme BCKDHA in melanoma pathogenesis and elucidated the underlying mechanisms. In vitro cell biology experiments and in vivo pre-clinical mice model experiments were performed to investigate the role of BCKDHA in melanoma progression. RNA sequencing, immunohistochemical/immunofluorescence staining and bioinformatics analysis were used to examine the underlying mechanism. BCKDHA expression was prominently increased in both melanoma tissues and cell lines. The up-regulation of BCKDHA promoted long-term tumour cell proliferation, invasion and migration in vitro and tumour growth in vivo. Through RNA-sequencing technology, it was found that BCKDHA regulated the expressions of lipogenic fatty acid synthase (FASN) and ATP-citrate lyase (ACLY), which was thereafter proved to mediate the oncogenic role of BCKDHA in melanoma. Our results demonstrate that BCKDHA promotes melanoma progression by regulating FASN and ACLY expressions. Targeting BCKDHA could be exploited as a promising strategy to restrain tumour progression in melanoma.

Abstract

Acne vulgaris (AV), a widely common disorder, that negatively affects the quality of life. Metformin is a relatively safe, cheap and well tolerated drug that is widely used in the treatment of Diabetes. Systemic metformin has demonstrated promising results in treating acne, while topically it was studied for melasma and recalcitrant central centrifugal cicatricial alopecia. To study the safety and efficacy of topical metformin 30% in the treatment of AV. Twenty-seven female AV patients were asked to blindly apply metformin and placebo gels to either side of the face for 12 weeks. AV lesion count was performed at baseline, at each visit and 4 weeks after end of treatment. At the end of the treatment period, the treated side showed significant improvement of comedones, papules and nodules but not pustules. Although, lesions count increased 1 month after stopping treatment, comedones and papules numbers were still significantly less on the metformin side compared to placebo. No side effects were reported. The limited number of patients studied and the limited follow-up period. The metformin effect was not studied on cellular and molecular levels. Topical metformin nanoemulsion gel can be a promising safe and effective treatment of AV.

Abstract

As extramammary Paget's disease (EMPD) sometimes invades and metastasizes from the skin to the mucosa, radical surgical resection of these lesions is often difficult. The purpose of this study was to analyse the association between surgical margins and survival as well as the benefit of functional preservation over complete resection, in patients with EMPD. We retrospectively analysed 230 patients diagnosed with EMPD between 1969 and 2020. Patient and treatment characteristics were recorded. Since our centre is a specialized hospital and almost all patients were referred from other hospitals, we reviewed their referral letters. Prognostic factors and survival time were also analysed. Among 230 patients, 78 (33.9%) had positive margins. The presence of margin positive lesions increased the local recurrence rate but was not significantly correlated with survival. Of all the patients who had received a thorough explanation about the surgical procedure in the referring hospital, 43.8% were scheduled for surgeries that would result in functional impairment, and all of them had function-preserving surgeries at our hospital with a 10-year survival rate of 100%. Our result suggest that less invasive surgery preserves anogenital and urethral function may be an acceptable option for EMPD treatment.

Abstract

The purpose of this study was to investigate whether hydrogen-rich bath has therapeutic effect on psoriasis and its molecular mechanism. Mice with imiquimod-induced psoriasis were established and divided into groups. The mice were respectively treated with hydrogen-rich water bath and distilled water bath. The changes of skin lesions and PSI scores of mice were compared after their treatments. HE staining was used to observe the pathological feature. The changes of inflammatory indexes and immune factors were analysed by ELISA and immunohistochemical staining. Malondialdehyde (MDA) content was measured by the thiobarbituric assay (TBA) method. By naked eye, the severity of skin lesions in hydrogen-rich water bath group was lower than that in distilled water bath group, and the psoriasis severity index (PSI) was lower (p < 0.01). The results of HE staining showed that the mice with distilled water bath had more abnormal keratosis, thickening of the spinous layer and prolongation of the dermal process, and more Munro abscess than the mice with hydrogen-rich water bath. During the course of disease, the overall levels and peaks of IL-17, IL-23, TNF-α, CD3⁺ and MDA in mice with hydrogen-rich bath were lower than those in mice with distilled water bath (p < 0.05). In the skin, the mice treated with the hydrogen-rich water bath also had lower peak of proliferating cell nuclear antigen (PCNA) levels. It is concluded that hydrogen-rich water bath can inhibit psoriasis inflammation and oxidative stress, relieve psoriasis skin lesions and accelerate the end of abnormal skin proliferation state, which shows a therapeutic and improving effect on psoriasis.

Abstract

Japanese patients with very high-risk cutaneous squamous cell carcinomas (cSCCs), based on the National Comprehensive Cancer Network guidelines, have been reported to display a higher cumulative incidence of relapse and disease-specific death (DSD) than those with high-risk cSCC. Therefore, prognosis prediction is crucial for Japanese patients with very high-risk cSCCs. Herein, we aimed to evaluate the prognostic prediction ability of our novel Japanese Risk Factor Scoring Systems (JARF scoring) in a Japanese cohort of cSSC patients. Data of 424 Japanese patients with resectable very high-risk cSCCs were analysed. We compared the prognostic ability of the following three staging systems: Brigham and Women's Hospital (BWH) tumour staging, number of NCCN very high-risk factors, and JARF scoring, including recurrent tumour, high-risk histological features, deep tumour invasion and lymphatic or vascular involvement as risk factors. The prognostic ability of these staging systems was evaluated according to the cumulative incidence of local recurrence (LR), regional lymph node metastasis (RLNM), DSD, and overall survival (OS). When BWH staging was used, high T stage led to significantly poor outcomes only in the cumulative incidence of RLNM (p = 0.01). The presence of very high-risk NCCN factors led to significantly poor outcomes in terms of RLNM (p = 0.03) and OS (p = 0.02). Meanwhile, a high number of risk factors in the JARF scoring system clearly led to poor outcomes in terms of LR (p = 0.01), RLNM (p < 0.01), DSD (p = 0.03), and OS (p < 0.01). The JARF scoring system may accurately predict the risk of recurrence and death in very high-risk cSCC patients in Japan.