The epididymis, a key reproductive organ, is crucial for sperm concentration, maturation, and storage. Despite a comprehensive understanding of many of its functions, several aspects of the complex processes within the epididymis remain obscure. Dysfunction in this organ is intricately connected to the formation of the microenvironment, disruptions in sperm maturation, and the progression of male infertility. Thus, elucidating the functional mechanisms of the epididymal epithelium is imperative. Given the variety of cell types present within the epididymal epithelium, utilizing a three-dimensional (3D) in vitro model provides a holistic and practical framework for exploring the multifaceted roles of the epididymis. Organoid cell culture, involving the co-cultivation of pluripotent or adult stem cells with growth factors on artificial matrix scaffolds, effectively recreates the in vivo cell growth microenvironment, thereby offering a promising avenue for studying the epididymis. The field of epididymal organoids is relatively new, with few studies focusing on their formation and even fewer detailing the generation of organoids that exhibit epididymis-specific structures and functions. Ongoing challenges in both clinical applications and mechanistic studies underscore the importance of this research. This review summarizes the established methodologies for inducing the in vitro cultivation of epididymal cells, outlines the various approaches for the development of epididymal organoids, and explores their potential applications in the field of male reproductive biology.

UNASSIGNED: Xeroderma pigmentosum (XP) is a rare inherited disorder with a high incidence of malignant tumours. Literature data on dermoscopic and in vivo reflectance confocal microscopy (RCM) findings in patients with XP are very limited.
UNASSIGNED: Dermoscopic findings in 32 biopsy-proven BCCs and RCM findings in 10 biopsy-proven BCCs developed in seven XP patients were reviewed.
UNASSIGNED: Of 32 BCCs, 28 were pigmented. On dermoscopy, BCCs exhibited multiple grey-blue globules/dots (81, 3%), short-fine telangiectasias/fine arborising vessels (65, 6%), multiple grey-blue ovoid nests (53, 1%), white structures (white-red structureless areas/shiny white areas/lines/strands) (56, 3%), arborising vessels (37, 5%), brown nests/globules/dots (28, 1%), spoke-wheel structures (9, 4%), leaf-like areas (9, 4%), ulceration (28, 1%), peripheral network (21, 9%), and multiple aggregated yellow-white globules (3, 1%). In 10 lesions in which further imaging with RCM was performed, RCM findings differentiated BCC from other tumours, including primary melanoma.
UNASSIGNED: Although the dominancy of pigmented structures may imitate melanoma clinically, dermoscopy is a valuable tool in the early diagnosis of BCCs in patients with XP. For suspicious lesions, RCM can help in differentiating pigmented BCC from primary melanoma.

The co-existence of an infiltrative basal cell carcinoma (iBCC) and a desmoplastic trichoepithelioma (DTE) within the same cutaneous lesion is a rare occurrence. iBCCs are relatively common malignant skin neoplasms that pose a risk for local tissue destruction and recurrence. DTEs are cutaneous neoplasms originating from hair follicles that may clinically and histologically appear similar to iBCCs but are ultimately benign. Distinguishing between these two entities is important given their differing destructive potential. Herein, we describe the case of a 36-year-old female with a single skin lesion on her left cheek that was comprised of both an iBCC distinct from a DTE, as verified by histopathologic analysis. A literature review highlights the rarity of such collision tumors and discusses the potential genetic links between these two histologically similar cutaneous neoplasms.

UNASSIGNED: The airway epithelium plays a central role in the pathogenesis of chronic respiratory diseases such as asthma and chronic rhinosinusitis with nasal polyps (CRSwNP), but the mechanisms by which airway epithelial cells (EpCs) maintain inflammation are poorly understood.
UNASSIGNED: We hypothesized that transcriptomic assessment of sorted airway EpCs across the spectrum of differentiation would allow us to define mechanisms by which EpCs perpetuate airway inflammation.
UNASSIGNED: Ethmoid sinus EpCs from adult patients with CRS were sorted into 3 subsets, bulk RNA sequenced, and analyzed for differentially expressed genes and pathways. Single cell RNA-seq (scRNA-seq) datasets from eosinophilic and non-eosinophilic CRSwNP and bulk RNA-seq of EpCs from mild/moderate and severe asthma were assessed. Immunofluorescent staining and ex vivo functional analysis of sinus EpCs were used to validate our findings.
UNASSIGNED: Analysis within and across purified EpC subsets revealed an enrichment in glycolytic programming in CRSwNP vs CRSsNP. Correlation analysis identified mammalian target of rapamycin complex 1 (mTORC1) as a potential regulator of the glycolytic program and identified EpC expression of cytokines and wound healing genes as potential sequelae. mTORC1 activity was upregulated in CRSwNP, and ex vivo inhibition demonstrated that mTOR is critical for EpC generation of CXCL8, IL-33, and CXCL2. Across patient samples, the degree of glycolytic activity was associated with T2 inflammation in CRSwNP, and with both T2 and non-T2 inflammation in severe asthma.
UNASSIGNED: Together, these findings highlight a metabolic axis required to support epithelial generation of cytokines critical to both chronic T2 and non-T2 inflammation in CRSwNP and asthma.

Rationale: IL-33 is a proinflammatory cytokine thought to play a role in the pathogenesis of asthma and chronic obstructive pulmonary disease (COPD). A recent clinical trial using an anti-IL-33 antibody showed a reduction in exacerbation and improved lung function in ex-smokers but not current smokers with COPD. Objectives: This study aimed to understand the effects of smoking status on IL-33. Methods: We investigated the association of smoking status with the level of gene expression of IL-33 in the airways in eight independent transcriptomic studies of lung airways. Additionally, we performed Western blot analysis and immunohistochemistry for IL-33 in lung tissue to assess protein levels. Measurements and Main Results: Across the bulk RNA-sequencing datasets, IL-33 gene expression and its signaling pathway were significantly lower in current versus former or never-smokers and increased upon smoking cessation (P < 0.05). Single-cell sequencing showed that IL-33 is predominantly expressed in resting basal epithelial cells and decreases during the differentiation process triggered by smoke exposure. We also found a higher transitioning of this cellular subpopulation into a more differentiated cell type during chronic smoking, potentially driving the reduction of IL-33. Protein analysis demonstrated lower IL-33 levels in lung tissue from current versus former smokers with COPD and a lower proportion of IL-33-positive basal cells in current versus ex-smoking controls. Conclusions: We provide strong evidence that cigarette smoke leads to an overall reduction in IL-33 expression in transcriptomic and protein level, and this may be due to the decrease in resting basal cells. Together, these findings may explain the clinical observation that a recent antibody-based anti-IL-33 treatment is more effective in former than current smokers with COPD.

UNASSIGNED: Full thickness defects of lower eyelid, following oncological resection, often pose a formidable challenge to the operating surgeon. Although a plethora of reconstructive options have been tried, the Mustardé flap has stood the test of time.
UNASSIGNED: Our patient, a 57-year old lady, diagnosed with basal cell carcinoma of the left lower eyelid underwent full-thickness wide local excision and reconstruction using modified Mustardé flap. The anterior part of defect was reconstructed using the flap. The posterior part was reconstructed using septal mucoperichondrial-cartilage composite graft, sutured to remnant palpebral conjunctiva.
UNASSIGNED: The patient had an uneventful post-operative period. On serial follow-up, the patient had an acceptable scar. At 6 months follow-up, the facial scar was barely perceptible with normal lower lid disposition.
UNASSIGNED: The modified Mustardé flap is a simple and reliable reconstructive option for full-thickness lower eyelid defects following oncological resection in carefully selected cases.

The immune system plays a key role in the suppression and progression of basal cell carcinoma (BCC). The primary aetiological factor for BCC development is exposure to ultraviolet radiation (UVR) which, particularly in lighter Fitzpatrick skin types, leads to the accumulation of DNA damage. UVR has roles in the generation of an immunosuppressive environment, facilitating cancer progression. Rates of BCC are elevated in immunosuppressed patients, and BCC may undergo spontaneous immune-mediated regression. Histologic and immunohistochemical profiling of BCCs consistently demonstrates the presence of an immune infiltrate and associated immune proteins. Early studies of immune checkpoint inhibitors reveal promising results in BCC. Therefore, the host immune system and tumor responses to it are important in BCC pathogenesis. Understanding these interactions will be beneficial for disease prognostication and therapeutic decisions.

Using scRNA-seq and microscopy, we describe a cell that is enriched in the lower airways of the developing human lung and identified by the unique coexpression of SCGB3A2/SFTPB/CFTR. To functionally interrogate these cells, we apply a single-cell barcode-based lineage tracing method, called CellTagging, to track the fate of SCGB3A2/SFTPB/CFTR cells during airway organoid differentiation in vitro. Lineage tracing reveals that these cells have a distinct differentiation potential from basal cells, giving rise predominantly to pulmonary neuroendocrine cells and a subset of multiciliated cells distinguished by high C6 and low MUC16 expression. Lineage tracing results are supported by studies using organoids and isolated cells from the lower noncartilaginous airway. We conclude that SCGB3A2/SFTPB/CFTR cells are enriched in the lower airways of the developing human lung and contribute to the epithelial diversity and heterogeneity in this region.

Basal cell carcinoma (BCC) is a major non-melanoma skin cancer, and its incidence is increasing worldwide. Although the main etiology is sun exposure, BCC may develop in sun-protected areas such as the vulva. The sonic hedgehog signaling pathway mutation may explain the mechanism underlying the occurrence of vulvar BCC. Owing to the rarity of metastases, wide local excision is an appropriate treatment option. Here, we report the cases two postmenopausal women with vulvar BCC who were surgically treated.

The present study aimed to evaluate the ability of a novel serum-free medium (SFM) to culture human airway epithelium cells (hAECs). hAECs were cultured in the novel SFM as the experimental group in the PneumaCult-Ex medium and Dulbecco\'s modified eagle medium (DMEM) and fetal bovine serum (FBS) as the control groups. Cell morphology, proliferative capacity, differentiation capacity and expression levels of basal cell markers were assessed accordingly in both culture systems. Optical microscope photos of hAECs were collected for cell morphology assessment. Cell Counting Kit-8 assay was conducted to assess the proliferation ability, and an air-liquid interface (ALI) assay was conducted to assess the differentiation capacity. Markers for proliferating basal and differentiated cells were relatively identified by immunohistochemical and immunofluorescent analysis. The results show that whether grown in the novel SFM or Ex medium, hAECs exhibited similar morphology at every passage, whereas cells could hardly form colonies in the DMEM + FBS group. Cells typically exhibited cobblestone shape, while a proportion of them in the novel SFM at late passage exhibited a larger shape. White vesicles appeared in the cytoplasm of some control cells at the later stage of culture. Basal cell markers (P63+KRT5+KI67+CC10-) for proliferating ability were found in the hAECs cultured by the novel SFM and Ex medium. hAECs at passage 3 cultured in the novel SFM and Ex medium both had the capacity to differentiate into ciliated cells (acetylated tubulin+), goblet cells (MUC5AC+) and club cells (CC10+) in the ALI culture assay. In conclusion, the novel SFM was capable of culturing hAECs. The hAECs cultured by the novel SFM could proliferate and differentiate in vitro. The novel SFM does not change the morphological characteristics or biomarkers of hAECs. The novel SFM has the potential for the amplification of hAECs for scientific research and clinical application.