Cancer cytopathologyGregor Barth, Stefan Koch, Thorsten H Ecke
BACKGROUND: Urine cytology is recommended for noninvasive detection of urothelial carcinoma (UC) but shows limited sensitivity, particularly in low-grade disease. Keratin 17 (K17) has emerged as a promising immunocytochemical marker. This study evaluated the diagnostic performance and interobserver agreement of K17 in a routine setting. METHODS: This prospective cross-sectional study included 184 urine samples from patients undergoing diagnostic evaluation for suspected UC. K17 immunocytochemistry was performed with a fully automated platform and independently evaluated by two blinded raters. Cytology was classified according to The Paris System for Reporting Urinary Cytology. UC status was determined by histology when available or by follow-up and multidisciplinary assessment. Diagnostic accuracy was assessed by receiver operating characteristic (ROC) analysis; interobserver reliability was assessed with the Cohen κ and intraclass correlation coefficient (ICC). Multivariable logistic regression evaluated combined diagnostic approaches. RESULTS: K17 expression was significantly higher in UC than in non-UC samples (p < .001). Across raters, area under the ROC curve (AUC) values ranged from 0.68 to 0.71. Sensitivity was 52.9%-65.9%, with a specificity of 73.7%-82.8%. Sensitivity ranged from 45.5% to 54.5% for low-grade tumors and from 57.7% to 73.1% for high-grade tumors. In stage-based analyses, sensitivity was 49.2%-61.5% for non-muscle-invasive UC and 65%-80% for muscle-invasive UC. Combining K17 with cytology numerically improved diagnostic discrimination (AUC value, 0.76; 95% CI, 0.69-0.83). Interobserver agreement was moderate to substantial (κ = 0.684; ICC, 0.653). Limitations included single-center design, partial histological verification (81%), and post hoc optimized cutoffs. CONCLUSIONS: K17 immunocytochemistry is a reproducible adjunct to urine cytology, with moderate diagnostic performance. It may warrant further evaluation as an adjunctive marker, particularly in low-grade and cytologically equivocal cases.
MedicineFengju Yuan, Qian Liu, Wuwu Ding, Jia Nie, Xianwei Wang
RATIONALE: Primary intravascular large B-cell lymphoma (IVLBCL) is a rare and aggressive extranodal lymphoma that rarely affects the prostate. Its nonspecific clinical and laboratory features often lead to misdiagnosis as benign prostatic hyperplasia (BPH) or prostatitis, delaying appropriate treatment. We report a case of primary prostatic IVLBCL initially misdiagnosed as BPH, highlighting the diagnostic challenges and the importance of comprehensive pathological evaluation. PATIENT CONCERNS: A 75-year-old male presented with a 1-year history of a weakened urinary stream, dribbling, increased nocturia, and urinary urgency. Symptoms transiently improved with self-medication of the α1-blocker tamsulosin but later recurred. DIAGNOSES: Histopathological examination revealed clusters of atypical tumor cells within the prostatic vasculature. Immunohistochemistry showed positivity for leukocyte common antigen, Vimentin, CD20, and CD79a, with a Ki-67 index > 90%. A final diagnosis of primary intravascular large B-cell lymphoma of the prostate was established. INTERVENTIONS: Following diagnosis, the patient and his family declined any antitumor therapy (including chemotherapy) and opted for best supportive care and were discharged against medical advice. OUTCOMES: The patient died 5 months after diagnosis without receiving any subsequent antitumor therapy. LESSONS: Prostatic IVLBCL is a diagnostic mimic of BPH and requires a high index of suspicion. Immunohistochemistry and molecular studies are essential for accurate diagnosis. Early recognition and appropriate chemotherapy can improve outcomes in this rare malignancy.
Oral and maxillofacial surgerySpriha Jha, Aadithya B Urs, Jeyaseelan Augustine, Priya Kumar
UNLABELLED: The diagnosis of primary bone tumours of the jaw often presents as a diagnostic dilemma. Overlap in clinical, radiological, and histological features between benign lesions like juvenile trabecular ossifying fibroma (JTOF); osteoblastoma (OB) and malignant tumours like low-grade osteosarcoma (LGOS) makes the diagnosis challenging. While osteosarcoma is a malignant tumour with poor prognosis requiring aggressive treatment, benign lesions have generally favourable outcome. This study aimed to assess the immunohistochemical expression of murine double minute 2 (MDM2) and Cyclin-dependent kinase 4 (CDK4) in low-grade osteosarcoma and benign osseous lesions. METHODOLOGY: Immunohistochemical analysis was performed on tissue samples from diagnosed cases of LGOS, JTOF, and OB. The expression levels of MDM2 and CDK4 were analysed and compared across the three groups. RESULTS: We found significantly higher expression of MDM2 and CDK4 in low-grade osteosarcoma compared to JTOF which showed negative expression in all of the cases. Osteoblastoma showed inconsistent and focal positivity contrary to OS where strong and diffuse expression of both the markers was observed. A strong co-expression of both the markers was also noted in 94% of OS cases. CONCLUSION: Immunohistochemistry for MDM2 and CDK4 is a valuable diagnostic tool in differentiating low-grade osteosarcoma from benign mimics. This approach provides a cost-effective, reliable, and accurate means for diagnosing jaw bone tumours, particularly when tissue samples are insufficient or on small incisional biopsies where accurate diagnosis is of prime importance.
Journal of cancer research and clinical oncologyM Weich, L Christl, M Kiesel, J Salmen, A Wöckel, J Diessner, E Gerhard-Hartmann, T Schlaiss
PURPOSE: The antibody-drug conjugates (ADCs) sacituzumab govitecan (SG) and datopotamab deruxtecan target trophoblast cell surface antigen 2 (TROP-2) and have shown significant efficacy in HER2-negative metastatic breast cancer (mBC). As TROP-2 may serve as a target across multiple treatment lines, detailed information on TROP-2 expression over time is of great interest. METHODS: TROP-2 expression was analyzed in breast cancer (BC) samples from patients treated at the University Hospital Würzburg between 2004 and 2025 at clinically indicated biopsy time points (TP), and, in a subset, before and after SG treatment. Expression was assessed immunohistochemically using the H-score (range 0-300). RESULTS: We evaluated 229 samples from 76 patients. Overall, patient-level TROP-2 expression was high (mean H-score 241.3 ± 73.2) and largely stable over time (TP1: 235.7 ± 60.8, n = 76; TP2: 244.6 ± 77.4, n = 72; TP3: 245.3 ± 84.4, n = 39; TP4: 244.9 ± 95.6, n = 8). However, 21 patients exhibited a decline in TROP-2 expression during disease progression, defined as a decrease of ≥ 50 H-score points between two consecutive TPs. Among 13 patients with paired biopsies obtained pre- and post-SG treatment, the mean TROP-2 H-score decreased from 227.0 ± 74.6 before SG to 202.9 ± 99.4 after SG (mean change - 24.1 ± 86.0; median 1.7, range - 170 to 110; Wilcoxon p = 0.542). TROP-2 H-score decreased in 6/13 patients and increased in 7/13 patients. Exploratory PFS analysis showed no statistically significant difference by post-SG TROP-2 change (log-rank p = 0.526). CONCLUSION: TROP-2 expression was generally high and relatively stable; however, some patients showed declining levels over time.
Pancreatic ductal adenocarcinoma (PDAC) exhibits diverse phenotypes, including epithelial and mesenchymal characteristics, yet these features have not been effectively translated into clinical applications. Mucins are implicated in tumor progression and therapeutic resistance and are considered potential diagnostic and therapeutic targets. In this study, five epithelial and three mesenchymal PDAC cell lines were cultured under two-dimensional (2D) and three-dimensional (3D) conditions to investigate mucin expression. Proteomic analysis identified five mucins (MUC1, MUC4, MUC5B, MUC19, and MUC20) in 2D culture and eight (including MUC2, MUC5AC, and MUC13) in 3D culture. Candidate mucins were further validated by immunocytochemistry with H-score assessment. MUC1 was consistently expressed in all PDAC cell lines and showed marked upregulation in several lines under 3D culture. In mesenchymal PDAC cell lines, mucin expression was largely restricted to MUC1, whereas epithelial lines displayed broad 3D-induced reorganization. Notably, MUC5AC was absent in 2D culture but robustly induced in all epithelial PDAC cell lines under 3D conditions. Other mucins, including MUC2, MUC4, MUC5B, MUC13, MUC19, and MUC20, were variably upregulated, with epithelial lines demonstrating higher diversity and intensity of expression. These findings demonstrate that 3D culture effectively reveals the plasticity and heterogeneity of mucin expression in PDAC, highlighting its potential as a platform for biomarker discovery and the development of therapeutic strategies.
Surgical and radiologic anatomy : SRAChristopher M Ahmad, Ambrose Loc Ngo, Rae-Anne Kastle, Phil Sheridan, Uyen Tam Nguyen, Melissa Zolnierz
BACKGROUND: Immersive virtual reality (VR) allows learners to interact with anatomical structures in three dimensions, potentially improving spatial understanding compared to traditional instructional methods. While VR is increasingly incorporated into medical curricula, prospective studies evaluating both learning outcomes and experiential measures remain limited. This study aimed to evaluate the impact of a single immersive VR anatomy session on knowledge acquisition, learner confidence, cognitive workload, and perceived immersion among first-year medical students. METHODS: In this prospective educational intervention, first-year medical students at Kansas City University completed a standardized immersive VR anatomy module. Anatomy knowledge was assessed using a 20-item multiple-choice examination administered pre-session, immediately post-session, and at a 20-day follow-up. Learner confidence was measured using a 5-point Likert scale. Cognitive workload was assessed using the NASA Task Load Index (NASA-TLX), and immersion was measured using validated presence and usability items. RESULTS: Anatomy knowledge improved significantly from pre- to post-intervention (mean increase 7.2% points, 95% CI [3.9, 10.6], p < 0.001). However, scores declined at delayed follow-up (mean change - 15.3% points from post, 95% CI [-19.8, -10.7], p < 0.001), falling below baseline mean levels. Confidence demonstrated a significant upward shift in distribution (p = 0.018), although change in confidence was not associated with knowledge gain (p = 0.863). Cognitive workload was not significantly associated with learning outcomes, while small negative correlations were observed between knowledge gain and both presence (r = -0.31, p = 0.022) and usability (r = -0.28, p = 0.034). CONCLUSION: Immersive VR produces significant short-term improvements in anatomy knowledge and learner confidence but does not sustain retention following a single exposure. While effective for enhancing short-term anatomy knowledge and learner engagement, these findings suggest that immersive VR may be most effective when integrated as a complementary component within a multimodal anatomy curriculum rather than used as a standalone instructional modality.
C-reactive Protein (CRP) has been established as a prognostic biomarker in various malignancies, with elevated serum levels correlating with poorer outcomes. However, the significance of tissue-based CRP expression specifically in chromophobe renal cell carcinoma (chRCC), remains inadequately characterized. This study investigates the potential prognostic relevance of intratumoral CRP expression from a substantial cohort of chRCC patients. We conducted a retrospective analysis of patients who underwent surgical intervention for chRCC. Comprehensive clinical data was collected, and immunohistochemical evaluation of tumor specimens was performed to assess intratumoral CRP expression patterns. The study included 81 chRCC patients, with intratumoral CRP expression identified in 35 cases (43.2%). Statistical analysis revealed no significant correlation between CRP expression status and clinical parameters. While 5-year overall survival (OS) analysis showed no statistically significant difference between CRP-positive versus CRP-negative tumors (86.4% versus 100.0%; p = 0.106), overall follow-up demonstrated a significantly higher mortality rate in patients with CRP-positive tumors compared to those with CRP-negative tumors (22.9% vs. 2.2%; p = 0.013). Our findings suggest that intratumoral CRP expression in chRCC is not clearly associated with parameters of aggressiveness or survival. Further studies should assess the possible correlation between CRP tissue expression and blood levels. However, these findings should be interpreted with caution given the limited sample size, low event rate, and high loss to follow-up, and are best considered hypothesis-generating.
Surgical and radiologic anatomy : SRASami Schranz, Hugues Cadas, Sara Sabatasso
PURPOSE: Intravascular injection of colored molding materials used either alone or in combination with radiological contrast agents is increasingly used to enhance vascular visualization in cadaveric specimens for anatomical education and surgical training. Published studies remain scattered and show substantial methodological variation. This structured literature review summarizes current practices regarding specimen preparation, injection techniques, casting materials, contrast agents, and imaging modalities, with particular emphasis on their educational applications. MATERIALS AND METHODS: A comprehensive literature search was conducted in PubMed, Scopus, Embase, Web of Science, and Google Scholar (1975-August 2025) using English, French, and German keywords. Reference lists were screened manually. After removal of duplicates, 702 records were screened, 145 full-text articles assessed, and 78 studies included. RESULTS: Included studies demonstrated variability in casting materials (primarily latex, with silicone, epoxy, and acrylic resins also reported), contrast agents (iodinated and barium-based compounds and lead oxide for CT, with gadolinium-based formulations predominantly used for MRI), perfusion pressures, flushing strategies, and specimen conditions (fresh-frozen, formalin-fixed, and Thiel-embalmed). However, methodological heterogeneity and inconsistent reporting limited direct comparison between approaches. The review nevertheless identifies representative reported technical protocols and highlights the main practical variables influencing protocol selection. CONCLUSION: Contrast-enhanced vascular casting provides valuable tools for anatomy education and surgical training by enabling direct correlation between radiological datasets and anatomical dissection. Beyond summarizing the literature, this review also offers practical protocol-selection guidance according to educational, research, and imaging objectives. As such, it may serve as a useful reference for teams implementing or refining these techniques in modern teaching environments.
Journal of molecular histologyAnsam M Z El Desoky, Heba Mahmoud Abdelgeleel, Manar Moustafa, Marwa Ahmed Mohamed Abdalrahman, Mai M Eldaly
Serous ovarian carcinoma (SOC) is frequently diagnosed at advanced stages and is associated with poor clinical outcomes, highlighting the urgent need for reliable diagnostic and prognostic biomarkers. Long non-coding RNA MEG3 and the autophagy-related protein Beclin-1 are recognized tumor suppressors; however, their combined diagnostic relevance in SOC remains insufficiently explored. This case-control study included 24 patients with histopathologically confirmed SOC. Paired tumor and adjacent non-tumorous ovarian tissues were analyzed for lncRNA MEG3 expression using quantitative real-time PCR and for Beclin-1 protein expression using immunohistochemistry. Serum CA-125 levels were assessed by ELISA. Associations with clinicopathological parameters were evaluated, and diagnostic performance was analyzed using receiver operating characteristic (ROC) curves. Both lncRNA MEG3 and Beclin-1 were significantly downregulated in SOC tissues compared with adjacent non-cancerous tissues (P < 0.0001). Reduced expression was significantly associated with tumor grade and ascites. The relationship with FIGO stage was not uniform in this cohort and should be interpreted cautiously because of the small sample size and unequal distribution of early and advanced cases. Beclin-1 expression was notably higher in premenopausal patients and in well to moderately differentiated tumors. A strong positive correlation was observed between lncRNA MEG3 and Beclin-1 expression (r = 0.96, P < 0.001), indicating a strong statistical association rather than a proven regulatory interaction. ROC curve analysis suggested diagnostic potential for lncRNA MEG3, Beclin-1, and serum CA-125. The concurrent downregulation of lncRNA MEG3 and Beclin-1 in SOC tissues suggests that these markers may serve as promising complementary biomarkers. However, their clinical utility should be considered preliminary and requires validation in larger, multicenter cohorts with functional studies before routine clinical application can be recommended.
Journal of molecular histologyHelena Ab'Sáber Simões, Cibele Pelissari, Giovanna Florezi, Ricardo Hsieh, Victor Elias Arana-Chavez, Cristina Massoco, Christine Delporte, Alexandre Ab'Saber,…
Salivary glands play an essential role in oral homeostasis by producing saliva, which protects oral tissues and maintains the oral environment. Despite growing interest in porcine models for translational biomedical research, the immunophenotypic characterization of porcine salivary glands remains limited in the literature, with few studies addressing their cytokeratin and contractile protein expression profiles. This gap constrains the ability to directly compare porcine and human glandular phenotypes and hinders the establishment of the pig as a validated salivary gland experimental model. This study evaluates the histological, immunophenotypic, and ultrastructural features of porcine submandibular glands as potential experimental models. Submandibular glands from 11 pigs aged 3 to 6 months (average weight: 20 kg) were examined histologically using hematoxylin and eosin staining and ultrastructurally by transmission electron microscopy, and phenotypically via immunohistochemistry for key markers such as CK5, CK7, CK19, SMA, calponin, caldesmon and S-100. Porcine submandibular glands exhibit a lobular organization akin to human glands, with mucous acinar cells, intercalated and excretory ducts, and rich vascularization. Immunohistochemistry revealed cytokeratins in epithelial cells and contractile proteins in myoepithelial cells, mirroring human glandular markers. Ultrastructural analysis highlighted robust cellular junctions, myoepithelial support, and intricate nerve fiber networks essential for glandular function. The structural and phenotypic parallels between porcine and human submandibular salivary glands provide a descriptive basis supporting their potential use in comparative salivary gland research.
Temperature is a key environmental driver of hepatic physiology in ectotherms, and three-dimensional (3D) fish liver models may serve as an ethically advantageous platform to investigate warming effects under controlled conditions while reducing the need for experimental animals. Despite their increasing use in toxicology, their application to assess climate-relevant temperature effects on liver function remains limited. This study investigated the temporal effects of a warming scenario on primary hepatocyte spheroids from juvenile brown trout (Salmo trutta), a bioindicator species. The study explores how a + 3 °C increase can affect spheroid development and maintenance, as well as its impact on metabolic activity, cell proliferation and death, and morphology over time. Spheroids were maintained at 18 °C and 21 °C for 25 days and analysed at five time points using metabolic, morphometric, immunocytochemical, and ultrastructural approaches. Mitochondrial metabolic activity, assessed by resazurin reduction, showed no significant temperature-related differences. In contrast, warming accelerated spheroid formation and produced larger spheroids. Proliferative activity, assessed by proliferating cell nuclear antigen (PCNA) immunostaining, was significantly reduced at 21 °C, while caspase-3 levels remained unchanged, indicating no increase in apoptosis. The autophagy marker microtubule-associated protein 1A/1B-light chain 3 (LC3A/B) showed lower immunoreactivity at 21 °C, with no temporal variation. Ultrastructural analysis revealed preserved hepatocyte integrity at both temperatures and abundant cytoplasmic dense bodies consistent with autolysosomal structures, which increased over time. Overall, a realistic warming scenario altered growth dynamics and cellular morphology. Further, this data reinforces that 3D fish liver models are viable alternative systems for assessing climate-driven effects.
Acta cirurgica brasileiraAnne Caroline Braz, Bruna Carla Hendges, Maria de Lourdes Pessole Biondo Simões, Sergio Ossamu Ioshii
PURPOSE: To compare the effects of captopril and losartan on inflammatory response, angiogenesis, and fibroblast activity during the early phase of wound healing in hypertensive rats. METHODS: Twenty-seven paraffin-embedded skin wound samples from hypertensive rats were analyzed: control group (n = 8), losartan-treated group (n = 9), and captopril-treated group (n = 10). Histological and immunohistochemical analyses were performed on postoperative day 4. Hematoxylin-eosin staining was used to evaluate general wound structure, and immunohistochemistry with anti-leukocyte common antigen, anti-CD34, and anti-smooth muscle actin was performed to assess inflammatory cells, angiogenesis, and fibroblast activity, respectively. RESULTS: No significant differences were observed in angiogenesis (p > 0.05) or fibroblast number (p > 0.05) between groups. However, the inflammatory reaction was significantly higher in captopril-treated (p = 0.0459) and losartan-treated (p = 0.0452) groups compared with the control group, with no significant difference between the two treated groups. CONCLUSION: Neither captopril nor losartan influenced angiogenesis or fibroblast proliferation in the early healing phase. However, both drugs increased inflammatory cell infiltration.
Microbial pathogenesisMarília Brasil Xavier, Lucas Dos Santos Fontes, Airton Kenji Motizuki, Leonardo Ramalho Mendonça Alves, Gabriela Fernandes Rodrigues, Larissa Dos Santos Alcânt…
BACKGROUND: Leishmania-HIV coinfection compromises the cutaneous immune response through mechanisms that remain poorly understood. This exploratory study compared the in situ immune microenvironment of skin lesions between coinfected and non-coinfected patients with American tegumentary leishmaniasis (ATL). METHODS: The in situ density of 20 immunohistochemical markers-covering cellular populations, regulatory and pro-inflammatory cytokines, and the inflammasome/pyroptosis axis-was quantified in skin biopsies from 23 patients with ATL: 17 without coinfection and 6 coinfected with HIV. Densities were compared between groups, stratified by highly active antiretroviral therapy (HAART), and explored by principal component analysis. RESULTS: The groups differed in sex distribution and clinical form, and no coinfected patient presented the localized form. Principal component analysis segregated the two immunological profiles. The ATL-HIV group showed reduced densities of CD1a+ dendritic cells, CD4+ and CD20+ lymphocytes, CD68+ macrophages, iNOS+ and CD163+ cells, as well as reduced IL-6+ and IL-1β+ cells. Conversely, FoxP3+, TGF-β+, IFN-γ+, and gasdermin D+ cells were increased. Patients on HAART remained clustered with the other coinfected patients, with no migration toward the ATL group. CONCLUSIONS: HIV is associated with a broad reorganization of the cutaneous immune microenvironment in ATL, combining depletion of effector populations, expansion of regulatory pathways, and engagement of alternative pyroptotic activity. These hypothesis-generating findings likely underestimate the functional impact of coinfection and warrant confirmation in larger studies.
Advances in physiology educationGregory J Crowther
NEW & NOTEWORTHY In a recent editorial, Erica Wehrwein warns that there is a shortage of people well prepared to teach in the disciplines of anatomy (A) and systems physiology (P). Here I elaborate on Wehrwein's concerns with a personal narrative on how I as a physiologist learned to teach anatomy. My pathway from a new A&P instructor to competent anatomist included three intermediate steps: 1) substituting P for A, 2) seeking help, and 3) evolving new structures.
Lung cancer (Amsterdam, Netherlands)Teodora Radonic, Sepehr Rahmani-Khajouei, Urania Dafni, Albrecht Stenzinger, Martina Haberecker, Line Bille Madsen, Kim Monkhorst, Jacek Kowalski, Javier Herna…
BACKGROUND: Functional NTRK1-2-3 fusions are rare, tumor-agnostic, targetable alterations. Accurate detection is essential to ensure optimal treatment. We investigated the prevalence of NTRK fusions in two cohorts of patients with NSCLC. METHODS: The Lungscape Cohort, comprising of tissue microarrays from 1703 resected stage I-III NSCLC cases (52% adenocarcinomas, 40% squamous cell carcinomas), was screened using pan-TRK immunohistochemistry (IHC, EPR17341, Ventana) and positive cases were confirmed by next-generation sequencing (NGS). The NGS Cohort of 24,243 NSCLC was screened by NGS within the European Lungscape Center Network and positive cases were retested using IHC. Functional fusions were defined as in-frame fusions containing an intact NTRK kinase domain. RESULTS: In the Lungscape Cohort of resected NSCLC panTRK IHC was found positive in 69 (60squamous cell carcinoma, 5 adenocarcinoma, 4 large cell carcinoma) of 1703 patients (4%, median H-score: 37, range 5-200). Sixty-five samples underwent confirmatory testing by NGS and one fusion was found, which after manual review was classified as non-functional. The NGS Cohort yielded 11 fusions in tumors of 24,243 patients of which eight were functional (0.03%; 5 in NTRK1, 1 in NTRK2 and 2 in NTRK3) and five were retested using IHC (median H score 200, range 50-300). Of the three non-functional fusions, two were IHC negative and 1 weakly positive (H-score 30). CONCLUSION: Pan-TRK IHC has a false-positive rate of ∼4%, particularly in squamous cell carcinomas, underscoring the need for confirmatory NGS to validate IHC findings and assess fusion functionality. Functional NTRK1/2/3 fusions remain exceedingly rare in NSCLC, with a detection rate of 0.03% across our combined cohorts.
International dental journalJing Hu, Ni An, Sheng Chang, Song Zhao, Rongjian Su, Jin Huang
OBJECTIVES: This study aimed to investigate the role of BMP9-Smad1/5/9 in the osteogenic differentiation and alveolar bone remodelling of Human periodontal ligament fibroblasts (hPDLFs) under the action of cyclic tensile stress (CTS). METHODS: An orthodontic tooth movement (OTM) model was established in Sprague-Dawley (SD) rats, the expression status of OCN and BMP9 was measured using immunohistochemistry. hPDLFs were subjected to CTS treatment (10% elongation, 0.5 Hz) to simulate the mechanical microenvironment during orthodontic treatment. The activity of ALP was detected by colorimetric assay and azo coupling staining, respectively. Mineralized nodules were visualized by Alizarin red staining. The expression levels of osteogenesis-related genes including RUNX2, ALP, and OCN were evaluated. To determine the role of BMP9-Smad1/5/9 in CTS-induced osteogenic differentiation, BMP9-Smad1/5/9 signalling was diminished by small RNA against BMP9 and the inhibitor of Smad signalling, the potential of osteogenic differentiation was assessed. RESULTS: BMP9 expression was upregulated in the tension-side periodontal ligament tissues of SD rats, as well as in hPDLFs stimulated by CTS. hPDLFs exhibited increased osteogenic differentiation potential under CTS induction, as evidenced by increased ALP activity, upregulated expression of osteogenesis-related genes, increased mineralized nodule formation and ALP staining. Mechanistically, CTS elevated the phosphorylation levels of Smad1/5/9. Silencing BMP9 attenuated CTS-induced osteogenic differentiation and reduced Smad1/5/9 phosphorylation. Moreover, inhibition of BMP9-Smad1/5/9 signalling pathway suppressed the osteogenic potential of hPDLFs. CONCLUSIONS: CTS increased the osteogenic potential of hPDLFs. Silencing of BMP9 and inhibition of the BMP9-Smad1/5/9 signalling inhibited the osteogenic differentiation of hPDLFs induced by CTS. CLINICAL RELEVANCE: These findings provide mechanistic insights into periodontal tissue remodelling and may contribute to the development of strategies to accelerate orthodontic tooth movement in clinical practice.
Annals of diagnostic pathologyYan Wang, Yeli Yao, Haiyan Shi, Weiguo Lu, Bingjian Lu
HPV-associated adenocarcinoma in situ (AIS) is the main precursor to endocervical adenocarcinoma. The differential immunoprofiles remain poorly characterized among HPV-associated AIS variants (such as usual-type and stratified mucin-producing intraepithelial lesion [SMILE]). This study aimed to investigate the expression of stem cell markers (SOX2 and SOX17) in 159 HPV-associated AIS (124 usual-type, 11 with intestinal differentiation, 24 SMILE) by immunohistochemistry. Compared to normal endocervical glands, AIS exhibited significantly increased expression of SOX2 (69.2% vs. 0, p < 0.001), but decreased SOX17 (40.9% vs. 98.1%, p < 0.001). Positive SOX2 and negative SOX17 had a specificity of 100% and 98.1% in the differential diagnosis of AIS from normal glands, respectively. SOX2 expression was significantly higher in SMILE (22/24, 91.7%) than in usual-type AIS (80/124, 64.5%, p = 0.030). In contrast, SOX17 expression was absent in all SMILE (0/24) and most AIS with intestinal differentiation (1/11, 9.1%), but was significantly higher in usual-type AIS (64/124, 51.6%, p < 0.001). The SOX2/SOX17 composite profile may aid in the diagnosis of AIS and its variants. The SOX2-positive (overexpression) /SOX17-negative profile suggest a divergent pathway with squamous differentiation potential in SMILE. These findings provide novel insights into the cellular histogenesis of AIS variants.
The involvement of necroptosis and the underlying mechanism in retinal ganglion cell (RGC) death is not fully understood. We aim to determine whether the NR_045396/miRNA761/Fas-associated protein with death domain (FADD) axis participates in the regulation of necroptosis in RGCs. A mouse model of optic nerve crush was employed for in vivo experiments. Apoptosis and necrosis were assessed by TUNEL and Propidium iodide (PI) exclusion. We found that the necrotic rate increased in a time-dependent manner in RGCs following optic nerve injury, whereas apoptosis peaked at 7 days following nerve damage. Immunohistochemistry revealed that the expression levels of key markers of necroptosis, pRIP3 and pMLKL, were upregulated, whereas FADD expression was reduced in RGCs at 14 days after optic nerve injury. Enforced expression of FADD in RGCs by an AAV vector attenuated necrotic response and promoted RGC survival. A dual-luciferase reporter gene assay showed that miR761 directly regulated FADD expression. Intraocular application of AAV2 expressing sequences complementary to miR761 binding site (AAV2-miR761 sponge) enhanced FADD expression and regulated RGC necrosis and survival. Moreover, the long non-coding RNA (lncRNA) NR_045396 binds directly to miR761 and modulates the necrotic program of RGCs. Thus, we demonstrate the anti-necroptosis and neuroprotective effects of the NR_045396/miR761/FADD axis.
Brain researchAlisha Braun, Phat Tai Vo, Larisa Bobrovskaya
The aim of this pilot study was to assess a time-dependent loss of orexin neurons in the unilateral intrastriatal 6-OHDA mouse model and to optimise the immunohistochemistry protocol for formalin-fixed, paraffin-embedded mouse brain tissue. A progressive loss of orexin A positive neurons in the lateral hypothalamus was observed in the lesioned side relative to the non-lesioned side. A significant reduction of orexin A positive neurons in the lateral hypothalamus was observed at 4 weeks with further loss at 6-8 weeks post 6-OHDA induction. Orexin neuron loss in the lateral hypothalamus has been reported in post-mortem Parkinson's disease patients and multiple animal studies, particularly in rats; however, evidence in neurotoxic mouse models remains limited. Orexin A positive neuron loss occurred alongside transient motor deficits, olfactory impairments, subtle cognitive changes and gastrointestinal dysfunction. Additionally, an orexin A immunohistochemistry protocol was successfully established for formalin-fixed, paraffin-embedded mouse brain tissue, enabling reproducible and reliable detection of orexin neurons as an alternative to frozen sections. This pilot study serves as a proof of concept that the unilateral intrastriatal 6-OHDA mouse model can successfully recapitulate the progressive loss of orexin A positive neurons and validates this model's suitably for investigating the longitudinal mechanisms of PD symptoms and pathology.
PathologyYijun Zheng, Yan Liu, Yan Ding, Zixiu Song, Xiaoya Zhao, AJin Hu, Lin Zhu, Jing Yang, Yuxiang Wang, Congrong Liu
SMARCA4-deficient uterine tumours encompass two distinct entities: SMARCA4-deficient undifferentiated/dedifferentiated endometrial carcinoma (SDUDEC) and SMARCA4-deficient uterine sarcoma (SDUS). Despite their divergent classifications, these tumours share overlapping clinicopathological and molecular features that complicate diagnosis. This study characterises their distinguishing features and clinical significance through analysis of 34 SDUDEC and 14 SDUS cases, integrating clinicopathological, immunohistochemical [BRG1 (encoded by SMARCA4), BRM (encoded by SMARCA2), INI1 (encoded by SMARCB1), ARID1A, ARID1B, SOX2, claudin-4, CK8/18, Chromogranin, synaptophysin, INSM1, p53, and MMR proteins], and next-generation sequencing (NGS; 14 SDUDEC and 14 SDUS) data. SDUDEC patients were significantly older than SDUS patients, with a median age of 54 years (range 28-63) compared to 37 years (range 24-58) (p<0.01). SDUDEC exhibited marked immunohistochemical heterogeneity, whereas SDUS showed uniform profiles: all SDUS cases (10/10) demonstrated dual BRG1/BRM loss with preserved expression of other SWI/SNF complex proteins, absent claudin-4 (0/12) and SOX2 (0/8) expression, and no MMR deficiency (0/14) or mutant-type p53 (0/13). NGS revealed frequent endometrial carcinoma-associated alterations in SDUDEC but rarely in SDUS. Prognostic outcomes did not differ significantly (p>0.05). These findings highlight distinct molecular landscapes: SDUDEC aligns with endometrial carcinogenesis, while SDUS may arise via alternative SMARCA4-dependent mechanisms. A diagnostic algorithm combining endometrial carcinoma molecular classification, SWI/SNF protein testing, and thorough sampling is proposed. This study expands the clinicopathological and molecular spectrum of SMARCA4-deficient uterine tumours, underscoring the need for entity-specific management strategies.