The Epstein-Barr virus (EBV) has two life cycles: latent and lytic. Previously, the latent cycle had been considered more relevant to EBV-associated epithelial malignancies, including nasopharyngeal carcinoma (NPC), whereas the lytic cycle, which ultimately leads to the death of host cells, had not been considered. However, recent studies have revealed that abortive lytic infections, in which the lytic cycle halts midway, avoids host cell death and contributes to tumorigenesis and tumor growth. However, the progression of the lytic cycle in NPC and its association with viral replication remain unclear. This study aimed to perform morphological analyses of EBV-positive NPC cell lines and clinical NPC biopsy samples using transmission electron microscopy, NanoSuit-correlative light and electron microscopy, and scanning ion conductance microscopy. Virion-like particles were observed in lytically induced EBV-positive NPC cell lines using time-lapse imaging to capture their release. These particles were also detected in clinical NPC samples using electron microscopy. Immunohistochemical analysis with the glycoprotein B (gB) antibody revealed a strong association between gB expression and poor prognosis. In conclusion, lytic infections with EBV replication occur in NPC, and the frequency of EBV replication is significantly associated with poor prognosis.

IntroductionIn human papillomavirus (HPV)-positive oropharyngeal cancer (OPC), lymph node metastasis is common, yet its biological and prognostic implications remain incompletely understood. Less is known about the spatial organization of immune programs within tumor-associated secondary lymphoid structures.MethodsImmune programs were analyzed by spatially distinguishing lymphoid follicular regions (LFRs) from tumor-free peritumoral tonsillar tissues (PTTs) in palatine tonsil–derived HPV-positive OPC. LFRs were analyzed using spatial transcriptomics and PTTs using bulk transcriptomics. Previously generated GeoMx Digital Spatial Profiler and bulk microarray datasets were reanalyzed to investigate compartment-specific immune programs. Immune pathway activity was assessed using single-sample gene set enrichment analysis according to nodal status.ResultsCompartment-specific immune programs associated with nodal status were identified in both LFRs and PTTs. In LFRs, nodal-positive cases showed enrichment of stem-like, pre-exhausted, and T-cell activity programs accompanied by inflammatory and interferon-associated signaling. In contrast, nodal-negative PTTs exhibited enrichment of stem-like, pre-exhausted, and T-cell activity programs together with IL2–STAT5 and TGF-β signaling, without overt inflammatory activation. These findings indicate that identical immune pathways exhibit distinct biological associations depending on their spatial localization.ConclusionImmune programs associated with nodal metastasis in HPV-positive OPC vary according to spatial immune context. Distinguishing LFRs from tumor-free PTTs provides a framework for interpreting immune programs beyond bulk tissue analysis and highlights the importance of spatial immune organization in shaping biological interpretation. Because the spatial transcriptomic analyses were performed in a limited number of patients, these findings should be considered exploratory and require validation in larger independent cohorts.

Background:Revision middle ear surgery for persistent conductive hearing loss after previous tympanoplasty or ossiculoplasty is technically demanding because of scar tissue, anatomical distortion, and the unpredictable status of the ossicular chain. Although transcanal endoscopic ear surgery (TEES) has been increasingly adopted for primary tympanoplasty and ossiculoplasty, its feasibility as the sole surgical modality in postoperative ears has rarely been reported. Case Presentation:We describe two adult patients (Case 1: a 63-year-old man and Case 2: a 72-year-old man) who presented with long-standing conductive hearing loss in dry, well-aerated postoperative ears following previous tympanoplasty performed at outside institutions. Preoperative four-frequency pure-tone average (PTA) air-bone gaps (ABGs) ranged from 28.75 to 40.00 dB (mean, 34.38 dB) with preserved bone-conduction (BC) thresholds. In both patients, TEES was performed exclusively through a minimal skin incision just above the tympanic membrane, without a retroauricular approach. Intraoperatively, fixation of the ossicular chain involved not only the previously placed columella but also the native ossicles, requiring complete release in each case. Type III tympanoplasty was performed in both cases. According to the IOOG SAMEO-ATO framework, both procedures were classified as S2r A1 Mx Ex Ox Ax Tx Ost. Outcome:The mean four-frequency PTA air-conduction threshold improved from 66.25 dB preoperatively to 42.19 dB postoperatively (mean hearing gain, 24.07 dB), and the mean postoperative ABG was reduced to 10.32 dB (range, 8.75-11.88 dB). Both patients (100%) fulfilled the criteria for postoperative hearing improvement according to the 2010 guidelines of the Japan Otological Society. No intraoperative or postoperative complications-including facial nerve injury, sensorineural hearing loss, dysgeusia, or vertigo-were observed during the follow-up period (mean, 18.5 months; range, 12-25 months). Conclusion:In carefully selected postoperative ears with a dry middle ear and good BC reserve, TEES may serve as a useful minimally invasive alternative to the conventional retroauricular microscopic approach, offering advantages in surgical visualization, operability, and reduced invasiveness. Larger studies are warranted to define optimal indications and long-term outcomes.

Background:Supralabyrinthine petrous bone cholesteatoma is a rare subtype in which hearing preservation may be achievable depending on the anatomical extent and surgical approach. Appropriate selection of surgical strategy is essential. Case Presentation:A 31-year-old man presented with a 2-year history of persistent left otorrhea. Otoscopic examination revealed pars flaccida cholesteatoma. Pure-tone audiometry demonstrated hearing loss with an air-bone gap of 35 dB. Computed tomography showed soft-tissue density extending from the tympanic cavity and mastoid air cells to the supralabyrinthine petrous apex. Inner ear function was almost preserved. Intervention:A staged surgical approach was adopted due to active inflammation. First, canal wall-down tympanomastoidectomy via a transmastoid approach was performed to control infection. Six months later, second-stage surgery was performed using the middle cranial fossa approach for complete removal of the supralabyrinthine cholesteatoma, along with ossicular chain reconstruction. Outcome:No intraoperative or postoperative complications occurred. Postoperative non-echo-planar diffusion-weighted MRI at 4 months showed no residual disease. At 22 months, hearing improved with an air-conduction threshold of 21.7 dB, and no tympanic membrane findings suggestive of recurrence were observed. Conclusion:In supralabyrinthine petrous bone cholesteatoma, selecting the surgical approach based on lesion extent, inflammatory status, and the need for hearing preservation is important for optimizing outcomes. In cases with extensive disease and significant inflammation, a staged strategy incorporating the middle cranial fossa approach may represent a safe and effective option for achieving disease eradication while minimizing the risk of inner ear dysfunction and postoperative complications.

Abstract Background Recurrent respiratory papillomatosis (RRP), caused by the human papillomavirus (HPV), is associated with an unpredictable clinical course. Although the Derkay Score is widely used to determine clinical severity, its prognostic value has rarely been evaluated. We developed a pathological severity score, the Hamamatsu Recurrent Respiratory Papillomatosis Pathological (HARRP) Score and combined it with the Derkay Score to develop a novel clinicopathological system—the Derkay-HARRP (D-H) Classification. We aimed to validate its prognostic value. Methods We retrospectively analyzed 125 patients who were clinically diagnosed with RRP from 16 Japanese institutions, randomly divided into validation (n = 38) and test (n = 87) cohorts. HPV-typing and immunohistochemistry for HPV-L1, HPV-E4, Ki-67, and p16 were performed. HPV particles were confirmed using the NanoSuit-correlative light and electron microscopy (CLEM) method. Receiver operating characteristic (ROC) curve analysis evaluated marker performance and defined recurrence cut-offs. HARRP and Derkay Scores were further assessed by ROC analysis and Cox proportional hazards models. We stratified patients using the D-H Classification and analyzed disease progression over time. Results No significant demographic differences were observed between the two cohorts. Positivity of HPV-L1, HPV-E4, and Ki-67 in the upper third of the epithelium was associated with recurrence. NanoSuit-CLEM confirmed HPV particles in HPV-L1–positive areas, supporting pathological relevance. The HARRP Score was calculated by assigning 1 point each for positivity of HPV-L1, HPV-E4, and ≥ 5% Ki-67-positive cells in the upper third of the epithelium. ROC analysis of the HARRP Score showed areas under the curve (AUCs) of 0.675 (validation) and 0.754 (test), whereas the Derkay Score showed AUCs of 0.709 and 0.834, respectively. The cut-off values were 1 and 4, respectively. Both scores were significant in the Cox analysis (p < 0.001). The D-H Classification stratified patients as Severe (both positive scores), Moderate (either positive), or Mild (both negative), with significant differences in relapse-free survival (p < 0.001). Severity tended to decrease with repeated surgeries and recurrence was rare in the Mild group. Findings remained consistent in HPV-positive cases only. Conclusions Combination of the Derkay and HARRP Scores—the D-H Classification—provides a practical tool for risk stratification and personalized follow-up planning of patients with RRP.

It is well known that a number of head and neck carcinomas are associated with HRAS mutations, and that several cancers with RAS mutations, such as lung cancer, have a poor prognosis. In this study, we evaluated the frequency of HRAS mutations in head and neck carcinomas and characterized the clinical and cell biological features of carcinomas with HRAS mutations. HRAS mutations at codons 12, 13, and 61, mutational hot spots, were evaluated in tissue specimens obtained from 119 Japanese patients treated at our institution. DNA was successfully extracted from 100 specimens, and sequencing was completed. An HRAS mutation was found in 8 (8.0%) cases: 5 (6.1%) out of 82 HNSCCs and 3 (16.7%) out of 18 salivary gland carcinomas. Mutations were found at codons 12 and 61, while none were found at codon 13, which differs from previous reports. The mutation-positive cases had a relatively poor prognosis, consistent with previous reports, and were more frequently accompanied by distant metastasis. HRAS knockdown with siRNA suppressed the in vitro migration ability of HRAS mutation-positive cells but not that of HRAS mutation-negative cells. In conclusion, a positive HRAS mutation could be an indicator of distant metastasis and poor prognosis, as well as a potential therapeutic target.

Objectives Nasal sinonasal inverted papillomas (SIP) are benign tumors with malignant potential that may progress to squamous cell carcinoma (SCC). Epidermal growth factor receptor (EGFR) expression is central to the development and malignant transformation of SIP, but the expression of its ligand, heparin-binding EGF-like growth factor (HB-EGF), and the regulators of HB-EGF expression have not been fully investigated. This study aimed to examine the expression of HB-EGF and its regulatory factors in SIP. Methods The expression of HB-EGF, EGFR, and phosphorylated EGFR (p-EGFR) was immunohistochemically examined in 32 patients with SIP, 3 with SIP-associated sinonasal squamous cell carcinoma (IPSCC), and 27 with de novo SCC arising in the sinonasal cavity (dnSCC). We also evaluated the expression of HIF-1α, an activator of ADAM12, which promotes the shedding of cytoplasmic membrane-bound HB-EGF from the cell surface. Results Among the 32 patients with SIP, most corresponded to Krouse stages T2-T3 and originated mainly in the maxillary or ethmoid sinus. All three cases of IPSCC were of the keratinizing type and classified as T4a. The 27 dnSCC cases were predominantly keratinizing type, arising chiefly in the maxillary sinus, and staged as T3-T4. HB-EGF and ADAM12 were expressed in all SIP and IPSCC cases, whereas the HB-EGF positivity rate was significantly lower in dnSCC (p < 0.05). EGFR and ADAM12 were broadly expressed across groups without significant differences, while p-EGFR expression was significantly higher in SIP and IPSCC compared with dnSCC (p < 0.05). Correlation analysis demonstrated a strong association between ADAM12 and HIF-1α, with weaker positive correlations observed between ADAM12 and HB-EGF, and between HB-EGF and p-EGFR. Conclusion HB-EGF expression is a characteristic feature of SIP. ADAM12 and HIF-1α may contribute to the distinctive upregulation of HB-EGF observed in SIP.

PURPOSE:Recurrent respiratory papillomatosis (RRP) is associated with low-risk types of human papillomavirus (HPV). With HPV DNA testing, the oral rinse of RRP patients may be a useful liquid biopsy, as previously shown in patients with oropharyngeal cancer. METHODS:Oral rinse, along with palatine and pharyngeal tonsil swabs, were collected from seven patients with persistent RRP. HPV DNA detection was performed using polymerase chain reaction, followed by genotype identification. RESULTS:HPV DNA was detected in five of seven oral rinse samples, but not in any palatine or pharyngeal tonsil swabs. HPV6 was identified in four of the five HPV-positive oral rinses, which was consistent with the RRP tissues. CONCLUSION:HPV DNA can be detected in oral rinses from patients with RRP, suggesting the utility of the oral rinse as a liquid biopsy. In contrast, neither the palatine nor the pharyngeal tonsils were reservoirs of HPV in study patients with RRP.

The Practical Guideline for the Management of Allergic Rhinitis in Japan was first published in 1993. After the COVID-19 pandemic, the current 10th edition was published in 2024. The most recent collection of evidence from the literature, such as the sustained post-treatment effect of sublingual immunotherapy on Japanese cedar pollinosis, was added to the revised guideline, which incorporates evidence-based medicine. In this revised guideline, a diagram illustrating the pathogenesis of allergic rhinitis and the mechanisms of action of various pharmacological treatments has been added. Also included is a diagram that shows the mechanism of action of allergen immunotherapy and a more detailed description of the oral allergy syndrome. The clinical question and answer section was also revised along with the introduction of new questions, such as: Does anti-IgE antibody treatment effectively reduce the symptoms of severe seasonal allergic rhinitis? Also updated was the evidence-based step-by-step strategy for treatment.

Background and Objectives: Japan’s aging population faces growing challenges related to oral frailty, a condition characterized by the decline of oral function associated with physical and nutritional deterioration. Impaired oral function contributes to reduced chewing, swallowing, and saliva secretion, leading to poor appetite and frailty progression. Ninjin’yoeito (NYT), a traditional Kampo formula, has been clinically used to improve systemic weakness and oral symptoms. This study aimed to evaluate the efficacy and safety of NYT in improving oral health among elderly individuals with impaired oral function. Materials and Methods: In this open-label prospective study, patients received NYT daily for 12 weeks. Assessments included oral symptom scores, mucosal moisture, repetitive saliva swallowing tests (RSST), gustatory function by visual analogue scale (VAS), an 11-item oral questionnaire, and immune profiling by flow cytometry. Safety was assessed through hematological and biochemical tests. Results: Symptom scores decreased from 8.27 at baseline to 3.64 at 12 weeks (p = 0.006), while oral condition scores improved from 5.09 to 1.36 (p = 0.006). Mucosal moisture increased (25.1 to 28.1, p = 0.03), and RSST frequency improved (2.18 to 4.55, p = 0.046). Questionnaire scores declined from 5.1 to 2.0 (p < 0.001). VAS-taste was unchanged overall (p = 0.21) but improved in low baseline patients. Laboratory findings showed no adverse changes, with favorable lipid trends. Immune analysis revealed a decline in NKG2D expression (p = 0.02), whereas other activating and inhibitory markers remained stable. Conclusions: NYT was well tolerated and associated with gradual improvements in oral and physical symptoms among elderly individuals with impaired oral function. These findings provide preliminary evidence supporting the feasibility of Kampo-based approaches for maintaining oral health in aging populations and warrant further validation in larger controlled trials.

Human papillomavirus (HPV) is a known risk factor for oropharyngeal cancer (OPC), with distinct HPV-positive and HPV-negative subtypes. Reactive oxygen species have been implicated in the carcinogenesis of several malignancies. Superoxide dismutase 2 (SOD2), a mitochondrial enzyme, is highly influenced by oxidative stress. This study investigated whether SOD1 and SOD2 expression in OPC affects primary tumor progression, lymph node metastasis, stage, and overall survival (OS). Biopsy or surgically resected specimens from 72 patients with OPC were analyzed via immunohistochemical staining for SOD1 and SOD2. The proportion of stained cells within the tumor area was assessed. Associations between SOD1 and SOD2 expression, T classification, N classification, and stage were evaluated. Factors correlated with OS in OPC were also examined. No significant differences in SOD1 or SOD2 expression were observed concerning T or N classification, or stage. However, high SOD2 expression was identified as a significant poor prognostic factor for OS. Regardless of HPV status, SOD2 expression predicts poor prognosis in OPC. Evaluating SOD2 expression may help predict OPC prognosis.

Background Cancer immune responses are generated in secondary lymphoid organs, such as the lymph nodes and tonsils. In the current study, transcriptional profiles of peritumoral tonsillar tissues (PTTs) from oropharyngeal cancers (OPCs) were assessed and compared with those of inflammatory tonsils and regional lymph nodes (rLNs). Methods RNA samples of PTTs and rLNs from 13 OPCs, and 4 inflammatory tonsils were subjected to microarray analysis, and differentially expressed genes (DEGs) identified from 730 nCounter Panel immune-related genes. Gene Set enrichment Analysis (GSEA) was used for DEG profiling of PTTs and rLNs between lymph node metastasis-negative and metastasis-positive cases. The top 20 genes, as ranked by GSEA metric scores, were extracted and subjected to principal component analysis (PCA). The correlation of each patient’s PCA score with lymph node status was assessed by Receiver Operating Characteristics (ROC) analysis. Results Comparing DEG analyses of PTTs with those of inflammatory tonsils and rLNs revealed 144 and 45 upregulated genes, respectively. ClueGO, a widely used Cytoscape plug-in, revealed activated pathways in PTTs, including lymphocyte proliferation (followed by T cell activation involved in the immune response) and positive regulation of leukocyte migration (followed by antimicrobial humoral immune response mediated by antimicrobial peptides) as the most significantly enriched immune system process functions in the gene ontology when comparing inflammatory tonsils and rLNs. The area under the ROC curves of PTTs and rLNs were 0.806 and 0.389, and were significant by DeLong’s test (p = 0.025). Conclusion PTTs exhibit unique immunological features distinguishing them from inflammatory tonsils and rLNs. Gene expression analysis of PTTs is useful for investigating the mechanism of OPC lymphatic spread, even compared with analysis of rLNs.

Conductive hearing loss typically results from ossicular chain abnormalities, commonly ossicular fixation or separation. While a precise diagnosis is useful for surgeons, distinguishing between fixation and separation before surgery is challenging. In our previous studies, we reported that sweep frequency impedance (SFI) effectively detects such middle-ear pathologies. However, due to the prolonged sound stimuli, SFI exhibited weaker resistance to noise. In this study, we introduce a novel method using short-time stimulation and adaptive noise reduction to improve SFI performance. The method was applied to both healthy individuals and patients, and a support vector machine was employed to evaluate its accuracy in distinguishing fixation and separation in clinical practice. The proposed SFI yielded results consistent with the original SFI meter but significantly shortened the evaluation time to within 200 ms. Classification results indicate that the SFI achieved accuracies of 98% and 83% for detecting ossicular separation and fixation, respectively. In contrast, such accuracies of traditional tympanometry were 70% and 49% for the separation and fixation. Additionally, the study indicates that gentle lullabies can serve as effective acoustic stimuli. These results suggest that our new SFI has potential for middle-ear testing across all age groups, from newborns to the elderly.

Background/Objectives: Juzentaihoto (JTT), a traditional Kampo formula composed of ten medicinal herbs, is widely prescribed in Japan for immune enhancement and general health maintenance. This exploratory, open-label pilot study aimed to evaluate the feasibility and immunomodulatory effects of JTT in cancer patients and to explore its potential mechanisms of action. Methods: Ten cancer patients received oral JTT (7.5 g/day) for 14 days, while healthy volunteers served as a reference group. Peripheral natural killer (NK) cell phenotypes and CD95 expression were analyzed by flow cytometry, and serum Fas ligand (FasL) concentrations were measured by ELISA. Complementary in vitro assays using PBS-extracted, autoclaved JTT were conducted to assess Fas/FasL-mediated apoptosis in Jurkat and primary T cells by flow cytometry and Western blotting for cleaved caspase-8 and -3. Additional experiments with staurosporine (intrinsic apoptosis) and TRAIL in OSC-19 carcinoma cells were performed to determine pathway specificity. Results: In patients, most NK-cell markers showed no statistically significant within-subject changes, although a trend-level increase in NKp46 and a significant reduction in NK-cell CD95 expression (paired p = 0.014) were observed. Between-group differences primarily reflected baseline disparities between cancer patients and healthy controls. In vitro, JTT (50–100 µg/mL) partially attenuated FasL-induced apoptosis and reduced cleaved caspase-3 without affecting cleaved caspase-8, suggesting selective downstream modulation of the extrinsic pathway. Conclusions: Within the limitations of a small, non-randomized cohort without placebo, these findings are hypothesis-generating and indicate that JTT selectively modulates Fas-mediated lymphocyte apoptosis without promoting tumor immune evasion. Further randomized trials and mechanistic studies incorporating co-culture or 3D tumor–immune models are warranted to confirm these observations and identify active constituents.

Sarcopenia is common in patients with head and neck cancer and is suggested to be associated with decreased survival. This study aimed to investigate the relationship between changes in skeletal muscle mass during alternating chemoradiotherapy (CRT) and the prognosis of patients with nasopharyngeal carcinoma (NPC). This retrospective study included 64 patients with NPC who had undergone alternating CRT at our institution between 2005 and 2022. The skeletal muscle mass index (SMI) was measured using pre- and post-treatment computed tomography. SMI decreased in 58 patients (90.6%), with a mean change of -6.1%. Using a cutoff value of -6.0% for SMI change, 32 patients (50.0%) were categorized into the SMI loss group. The SMI loss group had a significantly lower mean overall survival (OS) than the SMI maintenance group (122.6 vs. 153.0 months; p = 0.021). Multivariate analysis identified SMI loss and prognostic nutritional index (PNI) as independent predictors of poor OS (p < 0.05). They were used to construct the nomogram of OS. In conclusion, SMI loss during alternating CRT was identified as a poor prognostic factor. These findings suggest that preserving skeletal muscle mass during alternating CRT may improve the prognosis and merits further investigation.

Allergic rhinitis, especially Japanese cedar pollinosis, is considered a national affliction in Japan. The present exploratory study focused on the AllergoOncologic aspect of head and neck cancer (HNC) from the perspective of serum allergen-specific IgE against Japanese cedar pollen (JCP) for the first time. Serum JCP-specific IgE was measured in seventy-five HNC patients, including the oropharynx, hypopharynx, and larynx, and seventeen nasopharyngeal cancer (NPC) patients as well as forty-two patients with non-allergic benign disease (nABD). The JCP-specific IgE-positive rate was significantly lower in HNC than nABD (p = 0.037). Similarly, there was a clear trend toward a lower JCP-specific IgE score in HNC than nABD (p = 0.069). Further analyses excluding subjects in their 70s or older also showed that the JCP-specific IgE-positive rate was significantly lower in HNC than nABD (p = 0.022). In addition, The JCP-specific IgE score was significantly lower in HNC than nABD (p = 0.045). A lower serum JCP-specific IgE-positive rate in patients with HNC compared with nABD was demonstrated for the first time. An inverse correlation between HNC and Japanese cedar pollinosis was suggested from the perspective of allergen-specific IgE.

Nasopharyngeal carcinoma (NPC) is a malignant tumor in which the etiologic contribution of Epstein-Barr virus (EBV) is well established. However, similar to that of oropharyngeal carcinoma, some papers reported that human papilloma virus (HPV) contributed to the development of NPC in non-endemic regions. Previously, we conducted a study on HPV infection in patients with NPC between 1996 and 2015 in our department. The current study aims to evaluate the incidence and role of HPV infection in NPC pathogenesis using samples of NPC after 2015. Paraffin-embedded tumor samples from 26 patients with NPC who were treated at our department between 2015 and 2022 were analyzed. HPV polymerase chain reaction, p16 immunohistochemistry, HPV genotyping, and in situ hybridization for EBV-encoded RNA were performed to determine the viral infection status. Of the 26 patients, 19 (73%) were EBV-positive and HPV-negative, 1 (4%) was EBV-negative and HPV-positive, and 6 (23%) were negative for both EBV and HPV. The detection rate of HPV has slightly increased from 3% to 4% over the past decade. Although Japan is a non-endemic region for NPC, HPV infection is exceedingly rare and may have a limited role in NPC development in Japan. However, the detection rate of HPV has not significantly changed in the past decade, further supporting the view that HPV has a relatively small impact on the pathogenesis of NPC in Japan.