Penile squamous cell carcinoma (pSCC) is a rare cancer associated with a relatively poor prognosis. The WHO classifies pSCC into HPV-associated and HPV-independent subtypes, with HPV-positive pSCCs showing slightly better outcomes. A key negative prognostic marker is TP53 mutation. The aim of this study was to evaluate the concordance of (1) p16 immunohistochemistry with HPV status determined by PCR, (2) p53 immunohistochemistry with TP53 mutational status obtained by NGS, and (3) prognostic impact of overexpression and null phenotype p53 immunohistochemistry profiles. Analyzing invasive pSCC from 209 patients, we assessed concordance between p16 immunohistochemistry and HPV status (n = 132) determined by quantitative PCR, obtaining Cohen's kappa Κ = 0.62 (p < 0.0001). For p53 immunohistochemistry versus TP53 status by NGS (n = 145), Cohen's kappa reached Κ = 0.693 (p < 0.0001). We found worse overall survival (Hazard ratio = 3.33, 95% CIs 1.92-5.56, p < 0.0001) in p53 mutated cases (n = 38, both overexpression and null phenotype) compared to wild type (n = 171), but without a significant difference between both mutated profiles. Both HPV status and p53 profile can be reliably assessed by immunohistochemistry with substantial agreement. Patients with a mutated p53 profile showed significantly worse survival, irrespective of the mutated profile variant.
Objective: To investigate the diagnostic value of PRAME immunohistochemistry in the differential diagnosis between clear cell sarcoma (CCS) of soft tissue and melanoma, and its utility in routine diagnostic practice. Methods: A retrospective analysis was conducted on 25 CCS and 25 site-matched melanoma cases diagnosed at the Fudan University Shanghai Cancer Center from 2021 to 2025. Clinicopathologic data were collected, histologic features were assessed, and PRAME immunohistochemistry was performed. Clinicopathologic differences between the two groups were compared. The sensitivity and specificity of PRAME for differential diagnosis were evaluated. Results: Twenty-five patients with CCS were included, aged 38.0 (31.5, 56.0) years old. Another 25 patients with melanoma were included, aged 61.0 (54.0, 72.0) years old. In each group, 11 male and 14 female patients were included. Tumors were predominantly located in the extremities and trunks in both groups. In primary or recurrent melanomas, epidermal involvement with pagetoid proliferation at the dermoepidermal junction was frequently observed, whereas epidermal involvement was usually absent in CCS. Epithelioid morphology with prominent tumor-infiltrating lymphocytes and scant fibrous collagen septa were more often observed in metastatic melanomas, whereas a mixed epithelioid-spindle pattern, paucity of tumor-infiltrating lymphocytes, and frequent fibrous collagen septa were more commonly observed in CCS. PRAME negativity was observed in 96.0% (24/25) of the CCS cases, with only one case showing 5% nuclear positivity. By contrast, PRAME positivity was observed in 88.0% (22/25) of the melanoma cases, including diffuse nuclear positivity in>50% of tumor cells in 80.0% (20/25) of cases (P<0.001). With the cut-off of >50% positive cells, the sensitivity and specificity of PRAME for the diagnosis of melanoma were 80.0% and 100.0%, respectively. BRAF V600E immunohistochemistry was positive only in melanoma (56.0%, 14/25). All CCS cases were molecularly confirmed to harbor EWSR1 rearrangement.No co-occurrence of EWSR1 rearrangement and BRAF V600E immunohistochemistry positivity was observed in this cohort. Conclusions: Diffuse PRAME nuclear positivity is highly specific for diagnosing melanoma. It is particularly useful in biopsy specimens from recurrent lesions, needle biopsy specimens from metastatic lesions with inconspicuous pigmentation, biopsies of small primary lesions, and ulcerated specimens lacking an epidermal component. Therefore, PRAME immunohistochemistry is especially practical for community-hospital laboratories and interpreting biopsy specimens with limited tissue. 目的： 探讨PRAME在透明细胞软组织肉瘤（CCS）与黑色素瘤鉴别诊断及实际应用中的价值。 方法： 回顾性收集复旦大学附属肿瘤医院2021—2025年25例CCS与25例解剖学部位匹配的黑色素瘤，整理临床资料，评估组织学特征，行PRAME免疫组织化学检测。比较两组的临床病理学差异，并评价PRAME在其鉴别诊断中的灵敏度与特异度。 结果： CCS组25例，年龄38.0（31.5，56.0）岁；黑色素瘤组25例，年龄61.0（54.0，72.0）岁。50例患者中CCS组与黑色素瘤组均为男性11例，女性14例。肿瘤部位均以四肢与躯干为主。黑色素瘤原发灶/复发灶常累及表皮并见交界处Paget样增生，而CCS通常不累及表皮；当与黑色素瘤转移灶比较时，黑色素瘤多以上皮样细胞并伴明显肿瘤浸润淋巴细胞（TIL）且纤维胶原分隔少见，CCS多为上皮样及梭形的混合细胞型、缺乏TIL且纤维胶原分隔常见。免疫组织化学结果显示，CCS中96.0%（24/25）PRAME阴性，仅1例5%核阳性，而黑色素瘤中PRAME阳性率为88.0%（22/25），其中80.0%（20/25）呈>50%弥漫阳性（P<0.001）；以>50%为阈值，PRAME诊断黑色素瘤的灵敏度和特异度分别为80.0%和100.0%。BRAF V600E抗体免疫组织化学阳性仅见于黑色素瘤（56.0%，14/25）。入组CCS均经分子学证实存在EWSR1重排，且未见与BRAF V600E抗体免疫组织化学阳性共存。 结论： PRAME弥漫阳性对提示黑色素瘤具有很高的特异度，尤其在复发灶活检或转移灶穿刺且色素不明显时、原发小病灶的活检、或因溃疡未取到表皮的标本中，PRAME弥漫阳性更具提示意义，适合基层实验室及组织量有限的活检样本推广应用。.
The diagnosis of T-cell lymphoma is challenging, and establishing clonality is an important aspect of the workup. Molecular testing to establish T-cell clonality though T-cell receptor (TCR) gene rearrangement polymerase chain reaction is the most common modality and can detect clonal rearrangements in ∼90% of T-cell lymphomas, but is limited by increased turnaround time and cost and the need for sufficient material for testing. In the maturation of T cells, the TCR locus rearrangement will result in the mutually exclusive expression of either the TCR beta-chain constant domain 1 or 2 (TRBC1, TRBC2). Antibodies to TRBC1 and TRBC2 have been used in the setting of flow cytometry, and more recently immunohistochemistry, to establish T cell clonality. This study evaluates the utility of a TRBC1/CD3 dual stain for the diagnosis of T-cell lymphoma. The laboratory information system was searched from 2019 through 2025 for cytology samples with a diagnosis of T-cell lymphoma. Twelve samples were identified along with controls. Dual color immunohistochemistry for TRBC1 and CD3 was performed using a brown chromogen for TRBC1 and a red chromogen for CD3. The proportion of cells staining for TRBC1 and CD3 was scored. A monoclonal T-cell population was detected in all samples of T-cell lymphoma. All B-cell lymphomas and benign samples showed a polyclonal staining pattern consisting of a mixture of brown and red cells. Using immunohistochemistry for TRBC1 paired with CD3 is a reliable surrogate for T-cell clonality testing in cytology specimens and can support a diagnosis of T-cell lymphoma on limited material.
Immunohistochemistry (IHC) is a widely used technique for the identification, localization, and analysis of protein expression in tissues, playing a critical role in pathological diagnosis and validation of data derived from molecular biology. Despite its well-established use in mammalian tissues, applying IHC to fish still presents challenges, particularly due to the use of heterologous antibodies and the presence of paralogous genes. In this context, the present study aimed to standardize an immunohistochemistry protocol adapted for fish skeletal muscle using a biotinylated monoclonal antibody designed against a conserved region of the waslb gene for pacu (Piaractus mesopotamicus) and zebrafish (Danio rerio). Muscle samples were fixed in 4% paraformaldehyde, embedded in paraffin, and subjected to heat-induced antigen retrieval. After blocking nonspecific binding sites, histological sections were incubated with the primary antibody at an optimized 1:50 dilution, followed by detection with fluorophore-conjugated streptavidin (1:200) and nuclear counterstaining with DAPI. Reaction specificity was assessed using negative controls with a 20,000-fold molar excess of the immunizing peptide. The standardized protocol resulted in specific and reproducible labeling of the waslb protein in the skeletal muscle of both species, with no signal observed in negative controls; however, it is important to emphasize that this protocol can also be applied to antibodies against other fish genes. Thus, this method represents a reliable tool for protein analysis in fish muscle tissues, contributing to physiological, experimental, and comparative studies in non-mammalian models. © 2026 The Author(s). Current Protocols published by Wiley Periodicals LLC. Basic Protocol: Immunohistochemical reaction against the WASLB antibody in paraffin-embedded skeletal muscle sections of pacus and zebrafish.
Fetal growth restriction (FGR) is frequently defined as the failure of the fetus to reach its genetically predetermined growth potential. Heat shock proteins (HSPs) are extreme-temperature-resistant molecules that help proteostasis. The aim of this prospective case-control immunohistochemistry study is to evaluate the expression of HSP90 and HSP70 in the placentas of pregnancies complicated with FGR and compare their levels with the control placentas of normal-growth pregnancies. A prospective case-control study was conducted including people undergoing singleton pregnancies who gave birth in a tertiary university hospital in Central Greece. Participants were divided into two equal groups: an FGR pregnancy group and a control group with normal growth. Immunohistochemistry of placental samples was assessed using anti-HSP90 alpha/beta antibody (clone F-8, Santa Cruz Biotechnology, Dallas, TX, USA) and anti-HSC70/HSP70 antibody (clone W27, sc-24, Santa Cruz Biotechnology, Dallas, TX, USA). A scoring system was created to quantify the expression of HSP90 and HSP70 in each sample, and the grade of staining was measured at four points. A total of 80 pregnant people were prospectively enrolled in our study, with 40 in each group. Both constitutive (HSP90β and HSC70/HSPA8) and stress-inducible (HSP90α and HSP70/HSPA1A/B) isoforms were analyzed. When comparing the total score of HSP expression, a statistically significant difference was observed for both HSP90 and HSP70. For HSP90 expression, only the Hofbauer cell's stain was identified as a statistically significant independent factor, meaning that its positive expression was observed in Hofbauer cells. For HSP70 expression, only the staining of syncytiotrophoblasts was identified as an independent factor. FGR is a common pregnancy complication and a leading cause of stillbirth, neonatal mortality, and short- and long-term neonatal morbidity worldwide. Based on our findings, the lower expression levels of both HSP90 and HSP70 are associated with FGR, revealing a possible association with stress response in FGR pathophysiology. However, more robust data from larger-scale prospective studies are needed to elucidate the possible role of HSPs as potential FGR biomarkers.
Cholangiocarcinoma (CCA) remains a difficult-to-treat biliary malignancy in which therapeutic stratification increasingly depends on predictive biomarkers. Although next-generation sequencing is essential for the detection of targetable genomic alterations, immunohistochemistry (IHC) retains a practical role because it is widely available, rapid, and tissue-sparing. This narrative review summarizes the current and emerging role of predictive IHC in CCA, with emphasis on its clinical utility, interpretative pitfalls, and integration with molecular testing. HER2 and mismatch repair proteins currently represent the most relevant IHC-based markers in routine practice, albeit in selected subsets of patients. By contrast, PD-L1 has clear biological relevance but limited value as a stand-alone treatment selector in CCA, whereas Claudin 18.2 is promising but still investigational. Additional lines of research, including tumor microenvironment profiling, integrin-related pathways, and other theragnostic targets, may further refine patient selection, but these approaches are not yet standardized. Digital image analysis, radiomics, and machine learning are likely to improve quantification and may support future biomarker integration. A practical pathology-oriented approach should prioritize tissue stewardship, conservative interpretation of IHC results, and close coordination with molecular methods.
错配修复系统在维持基因组稳定性中发挥关键作用，其功能缺陷可导致微卫星不稳定，与多种肿瘤的发生、发展及免疫治疗反应密切相关。错配修复蛋白免疫组织化学（IHC）检测已成为评估错配修复状态的首选方法之一，对于Lynch综合征筛查、肿瘤分子分型、患者预后评估及免疫治疗决策具有重要意义。然而，目前国内错配修复蛋白IHC检测在标本前处理、抗体选择、检测系统、结果判读及质量控制等方面缺乏统一标准，导致不同实验室间检测结果存在差异。为此，中华医学会病理学分会联合技术学组、国家病理质控中心组织多学科专家，基于近年全国错配修复蛋白IHC室间质评数据、国内外循证研究结果及临床实践经验，形成本专家共识。本共识从检测意义、适用原则、检测流程、结果判读、报告规范及质控要求7个方面提出11条推荐意见，以期规范错配修复蛋白IHC检测流程、统一判读标准、强化质量控制，提高检测准确性，助力临床精准诊疗。.
To assess the frequency and level of expression of enhancer of zeste homologue 2 in cutaneous squamous cell carcinoma, and compare then with normal skin. The retrospective, descriptive study was conducted at the Department of Morbid Anatomy and Histopathology, University of Health Sciences, Lahore, Pakistan, and comprised data from January 2016 to April 2023 related to cutaneous squamous cell carcinoma in group A and normal skin cases in group B that were evaluated and compared for enhancer of zeste homologue 2 immunohistochemistry expression and positivity. Group A cases with increased immunohistochemistry expression were tested by fluorescence in situ hybridisation for enhancer of zeste homologue 2 amplification. Data was analysed using SPSS 22. Of the 60 patients, 30(50%) were in group A; 19(63.3%) males and 11(36.7%) females with mean age 48±16.5 years. There were 30(50%) patients in group B; 29(96.7%) females and 1(0.3%) male with mean age 47.6±11.3 years. There were 25(83.3%) well-differentiated cases in group A. Enhancer of zeste homologue 2 positivity was noted in 25(83.3%) case in group A compared to 6(20%) in group B (p<0.001). The enhancer of zeste homologue 2 expression level was also significantly higher in group A than group B (p<0.001). Immunohistochemistry overexpression of enhancer of zeste homologue 2 was found in 13(43.3%) group A cases, and, of them, 6(46.15%) showed enhancer of zeste homologue 2 amplification. Enhancer of zeste homologue 2 overexpression was noted in cutaneous squamous cell carcinoma cases, indicating that enhancer of zeste homologue 2 had a potential role in cutaneous squamous cell carcinoma tumourigenesis.
Postmortem human brains are important research resources to study the mechanisms underlying cerebrovascular features in various neurodegenerative disorders. Immunohistochemical and histochemical staining have been used to examine human brains fixed in neutral-buffered formalin (NBF) for months, years, or decades. Previously, we found that prolonged NBF fixation resulted in differential effects on immunohistochemistry and histochemistry staining on postmortem brains. Here, we further examined the effects of prolonged fixation (1-, 5-, 10-, 15-, 20-years) on stains of known biomarkers of cerebrovascular diseases in the human prefrontal cortex. We included microvasculature markers of the blood vessel wall (Anti-Collagen-IV and Claudin-5), a type III intermediate filament marker (Anti-Vimentin), an activated microglia marker (Anti-CD68), a biomarker for proteolipid protein (Anti-PLP) of oligodendrocytes and a marker for iron accumulation (Anti-Ferritin). We also included Masson's Trichrome Stain (MTS) and Bielschowsky silver stain (BSS). We found that staining intensities of Ferritin, Vimentin, Collagen-IV and BSS decreased with prolonged fixation. No significant differences were observed in the staining intensity of other markers. We therefore recommend performing IHC and HC staining for human brains with the same fixation times to offset any impact on downstream neuropathological analyses, as well as adding the fixation duration as a covariate in the analysis.
Orthotospovirus tomatomaculae (Tomato spotted wilt virus, TSWV) is an economically important plant pathogen reported to be transmissible by seeds. However, the pathways by which the virus enters the seed remain poorly understood. Here, we characterized the distribution and vertical transmission pathways of TSWV within reproductive organs of Capsicum annuum. Transmission electron microscopy (TEM) confirmed systemic distribution of TSWV throughout floral tissues, with virions detected in anthers, pollen grains, ovaries, ovules, and styles. Immunofluorescence histochemistry further revealed accumulation of the TSWV nucleocapsid (N) protein in the reproductive organs, with the strongest signals in anther septa, vascular bundles, and ovule integuments. Using reverse transcription-quantitative PCR (RT-qPCR) with an absolute standard curve, we detected the highest TSWV accumulation in anthers, ovaries, and styles, and lower levels in pollen grains. Cross-pollination assays demonstrated seed transmission via pollen (40%) and infected maternal tissue/ovules (30%), while self-pollination of infected plants resulted in 96% infected seeds. Testing of progeny seedlings showed infection rates of 20% (pollen-mediated), 30% (ovule-mediated), and 90% (self-pollination of infected parents). Together, these data support vertical transmission of TSWV through two reproductive routes, pollen and ovule, associated with providing a mechanistic framework for mitigating seed-borne spread in chili pepper.IMPORTANCETomato spotted wilt virus (TSWV) causes substantial yield losses worldwide, yet the mechanisms underlying its seed transmission remain insufficiently defined. We provide anatomical and molecular evidence that TSWV colonizes key reproductive tissues of Capsicum annuum, including anthers, pollen grains, ovaries, and ovules. Using controlled cross-pollination, we demonstrate that vertical transmission can occur through infected pollen (40% seed infection; 20% seedling infection) and through infected maternal tissues/ovules (30% seed infection; 30% seedling infection). Mechanistically, these data reveal two independent routes for vertical transmission: pollen-borne and ovule-borne. This finding directly supports two evidence-based management interventions: (i) the removal of infected male plants from seed production fields to block pollen-borne spread, and (ii) the selection of virus-free maternal lines to minimize ovule-mediated transmission. Incorporating both pollen- and ovule-associated transmission into epidemiological models and resistance breeding programs will therefore be critical for mitigating seed-borne TSWV losses in chili pepper.
Tongue morphology in cetaceans may reflect functional adaptations related to different feeding strategies. This study compared the lingual surface morphology of four delphinids species with different feeding strategies: the short-finned pilot whale (Globicephala macrorhynchus), Risso's dolphin (Grampus griseus), Clymene dolphin (Stenella clymene), and spinner dolphin (Stenella longirostris). Macroscopic dissections, light microscopy analysis, histochemistry, and scanning electron microscopy (SEM) were performed on four tongues, one from each species. The short-finned pilot whale and Risso's dolphin, which are suction feeders, exhibited thicker and rounder tongues, whereas the Clymene dolphin and spinner dolphin, which are raptorial prehension feeders, showed thinner and more elongated tongues. Across all species, only two types of lingual papillae, marginal and vallate, were identified, both lacking taste buds. Differences were also observed in the distribution of lingual glands, with suction feeders showing glandular concentration in the root of the tongue, while raptorial prehension feeders exhibited glands distributed along the body and root regions. Histochemical analysis revealed that serous cells showed positive staining with Periodic Acid-Schiff, whereas mucous cells stained positively with Alcian blue. SEM assessments indicated surface structures consistent with keratinocytes and glandular pores. These findings highlight functional tongue adaptations associated with distinct feeding strategies in delphinids.
Mucous membrane pemphigoid (MMP) is an autoimmune blistering disorder that can involve the esophagus, potentially leading to complications such as stricture or stenosis. Diagnosis of MMP is challenging owing to overlapping clinical features with other subepithelial blistering diseases and limitations of current diagnostic techniques. Although direct immunofluorescence (DIF) remains the gold standard, it requires fresh tissue samples, which are not always available, particularly in cases without active mucosal or cutaneous lesions. Complement fragment 4d (C4d) immunohistochemistry (IHC) on formalin-fixed tissue has demonstrated diagnostic utility in bullous pemphigoid but has been less explored in MMP. This case highlights that C4d IHC may serve as a useful ancillary test to support MMP diagnosis, especially for mucosal or esophageal biopsies received in formalin, offering a potential diagnostic pathway when fresh biopsy samples are unavailable.
Stenopodidea represents one of the basal lineages within Pleocyemata, yet the male reproductive system (MRS) of this group remains poorly understood, with limited information available regarding its morphology and function. This study provides the first detailed description of the MRS in four stenopodidean shrimp species from two families: Stenopodidae (Stenopus hispidus, S. scutellatus, and S. spinosus) and Spongicolidae (Microprosthema semilaeve). We analyzed the anatomy, histology, and histochemistry of the testes and vas deferens (VD), as well as spermatozoal ultrastructure, and compared these findings with data from more derived pleocyematans to identify potentially ancestral reproductive traits. Our analyses revealed two principal types of secretion in the VD of Stenopus species and three in M. semilaeve, which together form the presumptive spermatophore. Some secretions occur in small amounts and are restricted to specific VD regions. The layers surrounding the spermatozoa are relatively simple, consisting primarily of a sperm cord enclosed by thin secretory layers, suggesting a plesiomorphic reproductive condition in Stenopodidea. Spermatozoa are elliptical and characterized by a large nucleus in direct contact with the cytoplasm, numerous peripheral vesicles, and a large vesicle containing concentric membrane whorls. This structure, previously described as a lamellar body, is reinterpreted here as a putative acrosome vesicle and differs markedly from acrosome vesicles described in other Pleocyemata. Taken together, the comparatively simple spermatophore architecture and distinctive spermatozoa ultrastructure highlight Stenopodidea as an important lineage for understanding the early evolution of reproductive traits in Pleocyemata.
Ovarian tumors are classified into adenoma, borderline tumor, and adenocarcinoma based on cellular atypia and invasion. However, invasion can be difficult to assess histologically, underscoring the need for objective biomarkers. Laminin-5γ2 chain (Lam5γ2), a component of the basement membrane, has been implicated in tumor cell invasion in various malignancies. This study aimed to investigate the immunohistochemical expression of Lam5γ2 as a potential adjunctive biomarker associated with invasive phenotypes in ovarian serous and mucinous tumors. Lam5γ2 immunohistochemistry was performed on 80 ovarian tumor specimens, including adenomas, borderline tumors, and adenocarcinomas of serous and mucinous subtypes. Lam5γ2 expression patterns were compared across histological categories. In serous tumors, Lam5γ2 expression along the basement membrane was preserved in approximately half of adenomas and borderline tumors but was absent in all adenocarcinomas. Notably, cytoplasmic expression was observed in 11 of 12 serous adenocarcinomas (92%), and cytoplasmic staining was more frequent in borderline tumors and adenocarcinomas than in adenomas. A similar pattern was observed in mucinous tumors, with a decrease in basement membrane expression from adenomas to borderline tumors to adenocarcinomas. In mucinous adenocarcinomas, cytoplasmic expression was present in 6 of 10 cases, while basement membrane linear expression was retained in 3 of 10 cases. These differences were statistically significant in both serous and mucinous tumors (P <.001 and P = .03, respectively). Loss of Lam5γ2 expression from the basement membrane and aberrant cytoplasmic localization in ovarian serous and mucinous tumor cells were associated with histological progression and invasive phenotypes. Thus, Lam5γ2 immunoreactivity may serve as a potential adjunctive biomarker, particularly in diagnostically challenging borderline or microinvasive cases, pending validation in larger, independent cohorts.
Vascular cognitive impairment (VCI) shares major risk factors with heart failure with preserved ejection fraction (HFpEF), including obesity, diabetes and hypertension. Yet VCI research often relies on single-stimulus models, whereas patients experience combined risk factors. We therefore assessed cerebrovascular and cognitive phenotypes in an HFpEF model and investigated underlying mechanisms.Male Lean and Obese ZSF1 rats underwent longitudinal assessments of blood pressure, glucose, cardiac function, and behavioural performance. Cerebral blood flow and neurovascular coupling were assessed by laser speckle contrast imaging. White matter integrity, blood-brain barrier (BBB) permeability, and vascular density were analysed by (immuno)histochemistry. Cortical microvessels were isolated for transcriptomic profiling, and selected targets were validated using multiplex in-situ hybridization.Obese rats exhibited neurovascular uncoupling and impaired short- and long-term memory and spatial learning, accompanied by brain atrophy and reduced myelin. BBB permeability increased at 22-23 weeks and vascular density at 34-35 weeks in Obese vs Lean rats. Transcriptomic analysis of brain microvessels revealed altered processes related to angiogenesis, vasoreactivity, immune mechanisms and vascular remodelling, with consistent downregulation of Trpv4 and Klf2.Obese ZSF1 rats develop progressive neurovascular dysfunction associated with HFpEF onset and reduced Trpv4 and Klf2 expression in cerebral microvessels, two key vasoprotective genes.
Giant cell tumor of bone (GCTB) is a locally aggressive tumor driven by neoplastic mononuclear stromal cells. Stromal immunophenotyping may support diagnosis in giant cell-rich mimics and provide insight into tumor biology. Archival formalin-fixed, paraffin-embedded tissue samples from 31 GCTB cases diagnosed between 2015 and 2025 were studied and reviewed by consultant pathologists on-site and remotely to confirm the diagnosis and assess stromal atypia and mitoses. Immunohistochemistry for p63, cluster of differentiation 10 (CD10), and Ki-67 was performed and interpreted in stromal cells only; p63 and CD10 expression were categorized as low or high, and Ki-67 labeling index (LI) was grouped using a 10% cutoff. Available clinical variables included age, sex, anatomical site, tumor size, and recurrent-case status. Recurrent-case status was based on documentation at presentation or referral, not on prospectively confirmed recurrence. The mean age was 42.5 ± 15.4 years; 16/31 cases (51.6%) were female. Recurrent-case status was recorded in 14/31 cases (45.2%). High p63 and CD10 expression were observed in 20/31 cases (64.5%) and 22/31 cases (71.0%), respectively. In univariate analysis, CD10-high expression was associated with recurrent-case status (P = .021), while p63-high expression showed a borderline association (P = .057). Ki-67 LI was not associated with recurrent-case status (P = .389). p63 expression correlated significantly with CD10 expression (P = .003) and with higher Ki-67 LI. Stromal p63 and CD10 were commonly expressed in GCTB. CD10-high expression was associated with recurrent-case status in univariate analysis, but this exploratory finding should not be interpreted as recurrence prediction.
A series of studies have described aspects of outer-retinal morphology and visual opsins in different snake species. Among squamate reptiles, snakes stand out for their diversity in photoreceptor morphotypes and pronounced differences in retinal architecture between diurnal and nocturnal species. Diurnal snakes have a cone-dominated retina, lacking typical rods, but containing a transmuted, cone-like rod and exhibiting a low photoreceptor density. Nocturnal species have rod-dominated retinas with high rod densities and two or three types of cones. Despite these striking differences, no previous study has examined inner-retinal neuron types or aspects of neural circuitry in snakes. In this study, we used immunohistochemistry to investigate a specific type of bipolar cell immunoreactive to the anti-protein kinase C alpha (PKCα) antibody, a well-established marker for rod bipolar cells, and to assess its connectivity with photoreceptors in eight nocturnal and 13 diurnal species of snakes from eight families. Nocturnal snakes exhibited a canonical rod-selective PKCα-immunoreactive rod bipolar cell, whereas in diurnal species, the anti-PKCα antibody labeled a distinct population of long-wavelength-sensitive, cone-selective bipolar cells. This study reveals clear differences between diurnal and nocturnal species and highlights the remarkable plasticity and evolutionary innovation of the visual system in this group.
The unprocessed precursor of brain-derived neurotrophic factor (BDNF), proBDNF, has emerged as a potential determinant of therapeutic response in prostate cancer. Upon secretion, proBDNF preferentially binds to the co-receptors sortilin and p75NTR, triggering pro-apoptotic or pro-survival cascades, depending on cellular context. ProBDNF engages sortilin/p75NTR to drive castration resistance and metastasis in prostate cancer. High proBDNF/sortilin predicts poor therapy outcome, yet their tissue expression in prostate cancer (PCa) remains unclear. To evaluate the protein expression levels of proBDNF, sortilin, and p75NTR, we performed immunohistochemical analyses on 18 formalin-fixed paraffin-embedded (FFPE) PCa tissues obtained at radical prostatectomy between 2024 and 2025, together with matched para-carcinoma tissues. Compared with para-carcinoma tissues, immunohistochemistry in 18 paired specimens showed that pro-BDNF was significantly upregulated in PCa tissues (median IHC score 60.5 (range 57 - 65) vs. 41.5 (40 - 45), p < 0.05). Sortilin expression was also higher in PCa (median 37.0 (35 - 39) vs. 16.5 (15 - 18), p < 0.01); P75 expression remained relatively low in both prostate cancer and adjacent non-cancerous tissues (18.5 (17 - 20) vs. 6.5 (5 - 8), p < 0.05). These findings suggest a correlation between the expression levels of proBDNF, sortilin, and p75-NTR and the characteristics of PCa. Further investigation into the mechanisms underlying these interactions may provide valuable insights for the development of targeted therapies for PCa.
Sarcopenia has recently become a major public health issue. Adrenomedullin (AM) exerts diverse physiological effects, including angiogenesis, but its role in skeletal muscle is unclear. This study investigates whether AM can attenuate sarcopenic changes in aged mice compared to young adults. Young and old male C57BL/6J mice were randomly divided into two groups: the AM-treated group, which received subcutaneous administration of AM (50 nmol/kg body weight) 6 days a week, and the sham-treated group, which was injected with vehicle. After 3 months of treatment, we performed multi-muscle analyses (EDL, SOL, and PLA) using western blotting and immunohistochemistry. AM treatment significantly increased AM levels in plasma and muscle tissue. In young mice, AM was associated with changes in muscle fiber-type composition, including a reduction in Type IIx fibers, suggesting muscle-specific functional adaptation. In aged mice, AM increased CD31-positive capillary density and Pax7-positive muscle satellite cells (MuSCs), indicating improved vascular and stem cell niches. These changes occurred without alterations in muscle mass but were accompanied by improved motor function in aged mice. AM exerts age-dependent effects on skeletal muscle, with distinct responses observed in young and aged mice. These findings suggest that AM may contribute to muscle adaptation through context-dependent mechanisms involving vascular remodeling and fiber-type modulation.
Giant cell tumor of bone (GCTB) is a locally aggressive tumor with a considerable risk of recurrence, but reliable predictive markers are limited. High-temperature requirement A serine peptidase 1 (HtrA1) has been implicated in the progression of several cancers and may also play a role in GCTB recurrence. This retrospective study included 34 patients with GCTB treated with curettage between 2002 and 2016. HtrA1 expression was evaluated separately in giant cells (GC) and mononuclear cells (MNC) using immunohistochemistry. Based on high or low expression in each cell type, patients were categorized into four expression patterns: High GC / High MNC, High GC / Low MNC, Low GC / High MNC, and Low GC / Low MNC. Clinical data, including recurrence status and time to recurrence, were analyzed. The High GC / Low MNC group showed the highest recurrence rate at 71.4%, compared with 28.6% in the High GC / High MNC group and 31.6% in the Low GC / Low MNC group. No recurrence was observed in the Low GC / High MNC group, which included only one patient. The High GC / Low MNC group showed a non-significant trend toward higher recurrence compared with all other groups combined, with a similar trend toward shorter recurrence-free survival. HtrA1 expression patterns in GCTB may provide preliminary insight into recurrence risk. Although no statistically significant association was demonstrated, the observed trends suggest potential prognostic relevance and warrant validation in larger cohorts.