Toosendanin inhibits T-cell proliferation through the P38 MAPK signalling pathway.

In recent years, immunosuppressants have shown significant success in the treatment of autoimmune diseases. Therefore, there is an urgent need to develop additional immunosuppressants that offer more options for patients. Toosendanin has been shown to have immunosuppressive activity in vitro as well as effects on autoimmune hepatitis (AIH) in vivo. Toosendanin did not induce apoptosis in activated T-cells and affect the survival rate of naive T-cells. Toosendanin did not affect the expression of CD25 or secretion of IL-2 by activated T-cells, and not affect the expression of IL-4 and INF-γ. Toosendanin did not affect the phosphorylation of STAT5, ERK, AKT, P70S6K. However, toosendanin inhibited proliferation of anti-CD3/anti-CD28 mAbs-activated T-cells with IC50 of (10 ± 2.02) nM. Toosendanin arrested the cell cycle in the G0/G1 phase, significantly inhibited IL-6 and IL-17A secretion, promoted IL-10 expression, and inhibited the P38 MAPK pathway. Finally, toosendanin significantly alleviated ConA-induced AIH in mice. In Summary, toosendanin exhibited immunosuppressive activity in vivo and in vitro. Toosendanin inhibits the proliferation of activated T-cells through the P38 MAPK signalling pathway, significantly suppresses the expression of inflammatory factors, enhances the expression of anti-inflammatory factors, and effectively alleviates ConA-induced AIH in mice, suggesting that toosendanin may be a lead compound for the development of novel immunomodulatory agents with improved efficacy and reduced toxicity.

Hyperprogression of cutaneous T cell lymphoma after anti-PD-1 treatment.

BACKGROUNDImmune checkpoint blockade is an emerging treatment for T cell non-Hodgkin's lymphoma (T-NHL), but some patients with T-NHL have experienced hyperprogression with undetermined mechanisms upon anti-PD-1 therapy.METHODSSingle-cell RNA-Seq, whole-genome sequencing, whole-exome sequencing, and functional assays were performed on primary malignant T cells from a patient with advanced cutaneous T cell lymphoma who experienced hyperprogression upon anti-PD-1 treatment.RESULTSThe patient was enrolled in a clinical trial of anti-PD-1 therapy and experienced disease hyperprogression. Single-cell RNA-Seq revealed that PD-1 blockade elicited a remarkable activation and proliferation of the CD4+ malignant T cells, which showed functional PD-1 expression and an exhausted status. Further analyses identified somatic amplification of PRKCQ in the malignant T cells. PRKCQ encodes PKCθ; PKCθ is a key player in the T cell activation/NF-κB pathway. PRKCQ amplification led to high expressions of PKCθ and p-PKCθ (T538) on the malignant T cells, resulting in an oncogenic activation of the T cell receptor (TCR) signaling pathway. PD-1 blockade in this patient released this signaling, derepressed the proliferation of malignant T cells, and resulted in disease hyperprogression.CONCLUSIONOur study provides real-world clinical evidence that PD-1 acts as a tumor suppressor for malignant T cells with oncogenic TCR activation.TRIAL REGISTRATIONClinicalTrials.gov (NCT03809767).FUNDINGThe National Natural Science Foundation of China (81922058), the National Science Fund for Distinguished Young Scholars (T2125002), the National Science and Technology Major Project (2019YFC1315702), the National Youth Top-Notch Talent Support Program (283812), and the Peking University Clinical Medicine plus X Youth Project (PKU2019LCXQ012) supported this work.

Rare Concurrence of Alopecia Areata in the Setting of the Lipedematous Scalp.

ABSTRACT: The lipedematous scalp (LS) is characterized by a thickened scalp because of the increased thickness of the subcutaneous fat layer. When the soft and boggy scalp is associated with shortened hairs and hair loss, it is referred to as lipedematous alopecia (LA). We report a case of alopecia areata with LS, which may be misdiagnosed as LA. However, the histopathologic features showed a thickened subcutaneous fat layer and hair bulb inflammation. Hair regrowth was appreciable after treatment with topical steroids, minoxidil, oral compound glycyrrhizin, and vitamin D. This case report aims to show that LS and alopecia areata may coexist, and histopathologic examination is necessary for precise diagnosis.

Sis-25, a meroditerpenoid derivative with a cyclobutane scaffold, inhibits activated T cell proliferation by targeting GSK3ß in vitro and in vivo.

A series of novel scopariusicide derivatives were designed and synthesized starting from the main diterpenoid from the aerial parts of Isodon scoparius. Sis-25 was the most effective compound among them. The potential mechanism(s) of its immunosuppressive activity in vitro, as well as its effects on delayed type hypersensitivity (DTH) reaction and imiquimod-induced dermatitis in vivo were investigated in this study. Sis-25 inhibited anti-CD3/anti-CD28 mAbs, PHA or alloantigen-induced T cell proliferation without obvious cytotoxicity. Sis-25 was a highly selective inhibitor of GSK3-ß and inhibited the mTOR/p70S6K pathway but not the PI3K/Akt, p38 MAPK/ERK 1/2 and JAK3/STAT5 pathways. Furthermore, Sis-25 significantly inhibited IFN-γ, IL-6 and IL-17 expression but not IL-10 expression in activated T cells. Finally, Sis-25 treatment mitigated the DNFB-induced DTH reaction and ameliorated imiquimod-induced dermatitis. In summary, Sis-25 exerted significant immunosuppressive activity by targeting GSK3ß in vitro and in vivo. Sis-25 may guide the design of new drugs for more effective and safer treatments of autoimmune diseases and provide new insight into developing utilizations of Isodon scoparius.

Comparison Between Trichoscopic and Histopathological Evaluations of Hair Parameters.

Background: Collecting data on hair counts helps dermatologists diagnose hair diseases more accurately. Quantitative trichoscopy analysis and pathologic examination are two common ways to evaluate hair parameters. Purpose: The study aims to compare the effect of quantitative trichoscopy analysis and pathologic examination in hair counting and quantify the hair density of average Chinese people. Methods: Trichoscopy was performed in four scalp regions with a total of twenty-three volunteers without alopecia: frontal, vertex, occipital, and parietal. Hair density parameters were recorded. A 4-mm punch biopsy was performed in the occipital area, and all specimens were transversely sectioned serially and observed to calculate the hair densities. Results: The average hair density, hairs per follicle unit, and vellus hair ratio from quantitative trichoscopy analysis in the occipital region were 163.07±28.17/cm2, 1.87±0.25 and 6.60±3.95%, lower than those from biopsy significantly (214.97±48.73/cm2, 2.24±0.30 and 10.48±6.43%). The hair shaft diameter measured by quantitative trichoscopy analysis was 74.52±8.02µm, higher than that by histopathologic examination (68.65±8.00µm) (p<0.05). Conclusion: Quantitative trichoscopy is a noninvasive, quick, and convenient way to evaluate hair density. Histopathologic examination is an invasive way but provides more accurate data. The data obtained from trichoscopy and pathological examination are different to some extent, which cannot be substituted entirely for each other. This study established the reference for hair density in the adult Chinese population.

Concurrent extramammary Paget's disease involving the genitalia and axilla: Case report and literature review.

Concurrent multiple primary extramammary Paget's disease (EMPD) is rare. Herein, we present two Chinese cases of concurrent primary EMPD involving both the genitalia and the axilla, and they also had a history of other malignancy. We also summarise the cases of multiple primary EMPD previously described in literature. Careful examination of all apocrine sweat gland-bearing sites and additional internal malignancies is recommended for patients with EMPD.

Single-cell transcriptomics links malignant T cells to the tumor immune landscape in cutaneous T cell lymphoma.

Cutaneous T cell lymphoma (CTCL) represents a heterogeneous group of non-Hodgkin lymphoma distinguished by the presence of clonal malignant T cells. The heterogeneity of malignant T cells and the complex tumor microenvironment remain poorly characterized. With single-cell RNA analysis and bulk whole-exome sequencing on 19 skin lesions from 15 CTCL patients, we decipher the intra-tumor and inter-lesion diversity of CTCL patients and propose a multi-step tumor evolution model. We further establish a subtyping scheme based on the molecular features of malignant T cells and their pro-tumorigenic microenvironments: the TCyEM group, demonstrating a cytotoxic effector memory T cell phenotype, shows more M2 macrophages infiltration, while the TCM group, featured by a central memory T cell phenotype and adverse patient outcome, is infiltrated by highly exhausted CD8+ reactive T cells, B cells and Tregs with suppressive activities. Our results establish a solid basis for understanding the nature of CTCL and pave the way for future precision medicine for CTCL patients.

Prediction of surgical margin width using Mohs micrographic surgery for extramammary Paget's disease based on clinicopathological parameters: classification tree analysis of 150 cases.

Extramammary Paget's disease (EMPD) frequently extends beyond clinical borders, causing a high recurrence rate. Mohs micrographic surgery (MMS) has been used for management of EMPD, but its efficiency is compromised by technical limitations inherent in MMS. To identify clinicopathologic parameters of predictive value regarding MMS final margin width (FMW) for EMPD, and provide some preliminary guidance in selecting initial surgical margin width for improved efficiency. This was a retrospective study of 150 consecutive EMPD patients who underwent MMS between 2013 and 2019. Clinicopathological parameters and surgical data were collected to construct a classification tree of FMW. A six-node classification tree with a sensitivity of 86.25% and a specificity of 48.57% was generated. Lesion width, disease duration and inflammation score were used to select subgroups of patients in whom optimal initial margin width may be recommended. Classification tree analysis may help identify important variables to consider when selecting MMS initial surgical margins for EMPD.

Targeted inhibition of YAP/TAZ alters the biological behaviours of keloid fibroblasts.

Abnormal activation of fibroblasts plays a crucial role in keloid development. However, the mechanism of fibroblast activation remains to be determined. YAP/TAZ are key molecules in the Hippo signalling pathway that promote cell proliferation and inhibit apoptosis. Here, we show that keloid fibroblasts have higher levels of YAP/TAZ mRNA and proteins on primary culture. Targeted knockdown of endogenous YAP or TAZ significantly inhibited cell proliferation, reduced cell migration, induced cell apoptosis and down-regulated collagen1a1 production by keloid fibroblasts. Moreover, we demonstrate that verteporfin, an inhibitor of YAP/TAZ, has similar but stronger inhibitory effects on fibroblasts compared to YAP/TAZ knockdown. Our study provides evidence that YAP/TAZ may be involved in the pathogenesis of keloids. Targeted inhibition of YAP/TAZ could change the biological behaviours of fibroblasts and can potentially be used as therapy for keloids.

PEG10 amplification at 7q21.3 potentiates large-cell transformation in cutaneous T-cell lymphoma.

Mycosis fungoides (MF), the most common form of cutaneous T-cell lymphoma, undergo large-cell transformation (LCT) in the late stage, manifesting aggressive behavior, resistance to treatments, and poor prognosis, but the mechanisms involved remain unclear. To identify the molecular driver of LCT, we collected tumor samples from 133 MF patients and performed whole-transcriptome sequencing on 49 advanced-stage MF patients, followed by integrated copy number inference and genomic hybridization. Tumors with LCT showed unique transcriptional programs and enriched expressions of genes at chr7q. Paternally expressed gene 10 (PEG10), an imprinted gene at 7q21.3, was ectopically expressed in malignant T cells from LCT, driven by 7q21.3 amplification. Mechanistically, aberrant PEG10 expression increased cell size, promoted cell proliferation, and conferred treatment resistance by a PEG10/KLF2/NF-κB axis in in vitro and in vivo models. Pharmacologically targeting PEG10 reversed the phenotypes of proliferation and treatment resistance in LCT. Our findings reveal new molecular mechanisms underlying LCT and suggest that PEG10 inhibition may serve as a promising therapeutic approach in late-stage aggressive T-cell lymphoma.

Thyroid Hormone Receptor Agonist Promotes Hair Growth in Mice.

Background: Thyroxine is important to maintain the normal operation of the body. Both clinical and experimental results show thyroxine is closely related to hair growth, the mechanism of which is not fully understood. Purpose: Investigate the effect of thyroxine receptor agonist, TDM10842, for dorsal hair growth in C3H mice and explore its underlying mechanism. Methods: Depilated mice were applied with the TDM10842, vehicle of this drug and without any materials on dorsal skin. RNA-sequencing (RNA-seq) was employed to identify the change in gene expression of skin tissues. Quantitative real-time PCR (rt-PCR) and immunoblotting were conducted to validate key differentially expressed genes (DEGs) between different groups. Results: The TDM group showed early induction of anagen. 857, 782, and 276 differentially expressed genes were identified between 3 groups. As a critical DEG in group TDM, Pclaf was positively related to the motivation of Wnt/beta-catenin and Hedgehog signaling pathways, with a high expression of Ki67 and cyclinD1. Conclusion: TDM10842 accelerates the anagen entrance and the potential mechanism might be the activation of Wnt/beta-catenin and Hedgehog pathways. Pclaf serves as a critical molecule involved in pathway activation, and cyclinD1 is an important effector protein downstream of the pathways.

Effects of Penicillium expansum infection on the quality and flavor of yellow flesh kiwifruit during cold storage.

This study aimed to assess the effects of Penicillium expansum (P. expansum) infection on the quality and flavor of Jinmi (JM) and Jinyan (JY) kiwifruit. Kiwifruit were inoculated with P. expansum and stored at 0 ± 1°C, and the changes in quality indicators and volatile substances (VCs) at different stages of disease were determined. Results showed that in wound-inoculated kiwifruit, the soluble solid content (SSC) increased. Conversely, their titratable acidity and vitamin C (VC) content, firmness, lightness, and saturation decreased. The taste-related parameters and nutritional value of kiwifruit declined after infection. VCs such as ethanol, 3-methyl-1-butanol, and 2-methylisoborneol were detected only in the diseased fruit and gradually increased as the disease aggravated, suggesting that they may be the main sources of odor during P. expansum infection. Therefore, VCs detection can be used to determine possible P. expansum infection, as well as the degree of infection in kiwifruit. PRACTICAL APPLICATIONS: In practical application, we can use the results of this study to determine possible Penicillium expansum infection, as well as the degree of infection in kiwifruit according to the indicators such as volatile substances. Kiwifruit enterprises can use the nondestructive detection model established in this study to screen out the kiwifruit infected with P. expansum more efficiently, quickly, and accurately, in order to prevent harm to the health of consumers.

Hair Loss Caused by Gain-of-Function Mutant TRPV3 Is Associated with Premature Differentiation of Follicular Keratinocytes.

Gain-of-function mutations in the TRPV3 gene can cause Olmsted syndrome characterized by palmoplantar and periorificial keratoderma, itch, and hair loss. The mechanism underlying the hair loss remains unclear. In this study, we engineered an Olmsted syndrome mouse model by introducing the point mutation G568V to the corresponding Trpv3 locus in the mice. These mice developed fully penetrant hair loss. The hair loss was associated with premature differentiation of follicular keratinocytes characterized by precocious degeneration of trichohyalin and keratins, increased production of deiminated proteins, elevated apoptosis, and attenuation of transcription regulators (Foxn1, Msx2, Dlx3, and Gata3) known to regulate hair follicle differentiation. These abnormalities occurred in the medial‒proximal region of the inner root sheath and the hair shaft, where Trpv3 is highly expressed, and correlated with an impaired formation of the hair canal and the hair shaft. The mutant Trpv3 mice also exhibited increased proliferation in the outer root sheath, accelerated hair cycle, reduction of hair follicle stem cells, and miniaturization of regenerated hair follicles. Findings from this study suggest that precocious maturation of postmitotic follicular keratinocytes drives hair loss in patients with Olmsted syndrome.

Chinese Pre-service Music Teachers' Perceptions of Augmented Reality-Assisted Musical Instrument Learning.

Given the rapid growth of music technology, this study reports Chinese pre-service music teachers' perceptions of musical instrument learning assisted by augmented reality (AR). In this study, we conducted a small-scale case study with six pre-service teachers enrolled in a music teacher training programme at a comprehensive university in China. Participants engaged in semi-structured, face-to-face interviews after hands-on experiences with an AR-based piano learning app. Thematic analysis revealed that the participants were generally aware of the potential of this instructional approach but doubted its efficacy and exhibited weak intention to adopt it in their future classrooms. Implications of the findings for music teacher training are discussed.

A nomogram predicting subclinical extension of cutaneous squamous cell carcinoma in Chinese individuals: A retrospective study.

ABSTRACT: Cutaneous squamous cell carcinoma usually extends beyond the visible margin. Little is known about the predictors for cutaneous squamous cell carcinoma with subclinical extension in Chinese individuals. This study aimed to construct a nomogram for predicting the probability of subclinical extension of cutaneous squamous cell carcinoma in Chinese patients.A retrospective analysis was conducted using data from Mohs micrographic surgery-treated cutaneous squamous cell carcinoma patients at a single institution between December 1, 2009 and October 31, 2019. Subclinical extension was defined as a lesion requiring ≥ 2 Mohs stages or with final safe margins of ≥ 5 mm. A nomogram predicting the probability of subclinical extension was constructed using the predictors identified in multivariable analysis.Of 274 patients included, 119 (43.4%) had subclinical extension. In multivariable analysis, male sex (odds ratio [OR], 2.45; 95% confidence interval [CI], 1.40-4.29; P = .002), lesions on mucocutaneous areas (OR, 3.71; 95% CI, 1.34-10.32; P = .012) and extremities (OR, 2.40; 95% CI, 1.20-4.78; P = .013), maximum diameter of 10 to 19 mm (OR, 14.15; 95% CI, 4.24-47.28; P < .001), and 20 to 29 mm (OR, 9.21; 95% CI, 2.80-30.29; P < .001) were associated with subclinical extension. A nomogram incorporating these 3 variables demonstrated promising predictive ability (C statistics = 0.78; 95% CI, 0.67-0.89).The nomogram incorporating sex, tumor location, and maximum diameter can provide individualized prediction for subclinical extension in Chinese patients with cutaneous squamous cell carcinoma. This information may help surgeons determine appropriate margins at the first Mohs stage.