Artesunate and cisplatin synergistically inhibit HNSCC cell growth and promote apoptosis with artesunate­induced decreases in Rb and phosphorylated Rb levels.

In the treatment of head and neck cancer, cisplatin is often used as a therapeutic agent; however, its efficacy is limited and it can cause renal dysfunction as an adverse effect. For this reason, the use of cisplatin is limited in elderly patients with reduced renal function. Recently, artemisinin, which was developed as an antimalarial drug, was found to have antitumor effects and is effective in combination with other anticancer drugs. In the present study, the antitumor effects of artemisinin and its derivatives as well as their combination with cisplatin and iron on head and neck squamous cell carcinoma cell lines, were investigated. Cell viability was determined by a cell viability assay, the cell cycle was analyzed by flow cytometry, cell death was assessed with annexin V and propidium iodide staining, and western blotting was used to analyze retinoblastoma protein (Rb), phosphorylated (p­)Rb, and other cell cycle­associated molecules. A total of four artemisinin compounds were examined and it was found that artesunate and dihydroartemisinin had a significant inhibitory effect on growth. It was also identified that the combination of artesunate, cisplatin, and iron inhibited cell proliferation and caused S/G2­M cell cycle arrest. In addition, western blotting of Rb, a molecule involved in the cell cycle, showed that artesunate induced the loss of not only Rb but also p­Rb. These results suggested that artesunate is a useful drug in combination with cisplatin.

A Novel Treatment Strategy for Advanced Lacrimal Sac Carcinomas Confirmed by p16 Immunostaining.

This article aims to describe the two cases in which chemotherapy and chemoradiotherapy were effective for advanced HPV-related lacrimal sac squamous cell carcinoma and avoided the need for radical surgery. This was an interventional study of two patients with advanced lacrimal sac squamous cell carcinoma. Two patients with advanced lacrimal sac squamous cell carcinoma were treated at our University Hospital between January 2020 and February 2021. Diagnosis of HPV-related lacrimal sac carcinoma was done by p16 immunostaining and RNA in situ hybridization. Received neoadjuvant chemotherapy and chemoradiotherapy, also minimally invasive surgery to remove any residual tumor if the final response, were unfavorable. HPV-related carcinoma was decided by checking p16 and RNA status. Response was assessed by computed tomography, magnetic resonance imaging, positron emission tomography-computed tomography, and endoscopic images. Both patients had positive p16 staining also HPV RNA in situ hybridization. Received definitive chemoradiotherapy instead of radical surgery after showing a partial response to neoadjuvant chemotherapy. A complete response was achieved in one patient and the other had a partial response, leaving a small residual tumor in the nose that was successfully removed by endonasal endoscopic surgery. Cure was achieved in two patients with HPV-related lacrimal sac squamous cell carcinoma by neoadjuvant chemotherapy followed by definitive chemoradiotherapy, with only one requiring minimally invasive surgery. This is a new direction in the treatment of p16-positive lacrimal sac carcinoma, especially for advanced cases, whereby molecular biological indicators can be used to avoid highly invasive surgery and preserve quality of life without compromising prognosis.

Identification of cisplatin-resistant factor by integration of transcriptomic and proteomic data using head and neck carcinoma cell lines.

Cisplatin is an important drug for the treatment of head and neck squamous cell carcinoma (HNSCC). Determining chemoresistant factors prior to treatment will lead to great benefits for clinicians and patients. Here, we evaluated chemoresistant factors by integrating proteomic and transcriptomic data using HNSCC cell lines to identify a more precise chemoresistant factor in HNSCC. We used four HNSCC cell lines: cisplatin-sensitive, acquired cisplatin resistance, naturally cisplatin-resistant, and acquired 5-FU resistance. Proteomic analysis was performed using iTRAQ, tandem mass spectrometry, and liquid chromatography-electrospray ionization-tandem mass spectrometry. Transcriptomic analysis was performed using microarrays. By integrating these independent data, common factors were addressed and functional analysis was performed using small interfering RNAs (siRNAs) to change the chemosensitivity. Using iTRAQ analysis, 7 proteins were identified as specific for cisplatin chemoresistance factors. Transcriptomic analysis revealed hundreds of potential candidate factors. By combining and integrating these data, S100A2 was identified as a potential cisplatin-specific chemoresistance factor. Functional analysis with siRNA revealed that the expression of S100A2 was reduced and cisplatin sensitivity recovered in the acquired and naturally cisplatin-resistant cell lines, but not in the cisplatin-sensitive cell lines. S100A2 was identified as a cisplatin-specific chemoresistance factor by integrating the transcriptomic and proteomic results obtained using HNSCC cell lines. This is a novel technique that allows for a precise identification, also known as a comprehensive analysis. Our findings indicate that these proteins could be used as biomarkers of HNSCC treatments, providing physicians with new treatment strategies for patients with HNSCC, showing chemoresistance.

Incisionless facial resection for Kadish stage C olfactory neuroblastoma: Transcaruncular approach with combined endonasal and skull base surgery.

This case report describes resection without facial incision for aggressive Kadish stage C olfactory neuroblastoma (ONB). We performed resection via transcaruncular approach with combined endonasal and skull base surgery. This multidisciplinary team surgical approach is expected to lead to a new strategy for this type of tumor in the future.

Immune-checkpoint molecules on regulatory T-cells as a potential therapeutic target in head and neck squamous cell cancers.

Immune-checkpoint inhibitors improve the survival of head and neck squamous cell carcinoma (HNSCC) patients. Although recent studies have demonstrated that the tumor immune microenvironment (TIME) has critical roles in immunotherapy, the precise mechanisms involved are unclear. Therefore, further investigations of TIME are required for the improvement of immunotherapy. The frequency of effector regulatory T-cells (eTregs) and the expression of immune-checkpoint molecules (ICM) on eTregs and conventional T-cells (Tconvs) both in peripheral blood lymphocytes (PBL) and tumor-infiltrating lymphocytes (TIL) from HNSCC patients were analyzed by flow cytometry and their distributions were evaluated by multi-color immunofluorescence microscopy. High frequency eTreg infiltration into HNSCC tissues was observed and high expressions of CD25, FOXP3, stimulatory-ICM (4-1BB, ICOS, OX40 and GITR) and inhibitory-ICM (programmed cell death-1 [PD-1] and cytotoxic T-lymphocyte-associated protein-4 [CTLA-4]) were found on invasive eTregs. In contrast, the expression of stimulatory-ICM on Tconvs was low and the expression of inhibitory-ICM was high. In addition, ICM-ligands (programmed cell death-1 [PD-L1], galectin-9 and CEACAM-1) were frequently expressed on cancer cells. PD-L1 and galectin-9 were also expressed on macrophages. PD-1+ T-cells interacted with PD-L1+ cancer cells or PD-L1+ macrophages. This suggested that in TIL, eTregs are highly activated, but Tconvs are exhausted or inactivated by eTregs and immune-checkpoint systems, and ICM and eTregs are strongly involved in the creation of an immunosuppressive environment in HNSCC tissues. These suggested eTreg targeting drugs are expected to be a combination partner with immune-checkpoint inhibitors that will improve immunotherapy of HNSCC.

Thyroid Gland Flap for Minimally Invasive Reconstructive Head and Neck Surgery.

Head and neck surgery sometimes causes small defects, and salvage surgery after chemoradiotherapy poses some risk because of damage to the surgical site from the previous treatment. We have developed a novel thyroid gland flap for head and neck surgical reconstruction and here we describe elevating the flap, including arc rotation, size, and suture technique, and our outcomes to date. METHODS: Thyroid gland flap reconstruction was performed in 13 cases (11 patients) between July 2009 and May 2020. The clinical importance and adverse effects of the procedure were examined. Thyroid function and blood flow of the flap were assessed, and the status of the flap and irradiated recipient tissue was examined histopathologically. RESULTS: Median age at surgery was 64.6 years (range 49-77 years). Two of the patients underwent reconstruction with a thyroid gland flap twice. There were 4 cases of primary head and neck cancer resection with neck dissection in which the flap was harvested from the thyroid gland as reinforcement. In 1 case, surgery was performed for cervical esophageal diverticulum. In all cases, the arc was limited to 6 cm and suturing was basic. There were no complications of the surgical procedure, and the postoperative course was uneventful. Contrast-enhanced computed tomography revealed adequate enhancement of the flap. Postoperative thyroid function was normal. The thyroid gland flap was firmly adapted and fused with the irradiated recipient tissue. CONCLUSION: The thyroid gland flap could be an effective tissue flap fed by the superior thyroid arteriovenous pedicle for head and neck reconstruction.

Thyroid gland flap minimizes mucosal defects at supracricoid partial laryngectomy with cricohyoidoepiglottopexy.

Wound infection is a major complication after supracricoid partial laryngectomy with cricohyoidoepiglottopexy (CHEP) for radiation therapy failure. A 60-year-old man received chemoradiotherapy for a glottic carcinoma. CHEP, reusing the thyroid gland flap (TF), was performed because the cancer recurred after a salvage vertical partial laryngectomy following radiation therapy failure. The TF was sutured to the supraglottis and cricoid cartilage mucosa to minimize mucosal defects before the hyoid bone and cricoid cartilage were sutured. Wound healing after CHEP was good without infection. After decannulation, oral food intake was possible without aspiration, and speech function was comparable to that of other patients who had supracricoid partial laryngectomies. Histopathological examination revealed a close connection between the TF and its surrounding tissues without fibrous scarring. TF may improve wound healing after CHEP for radiation failure by minimizing mucosal defects.

A novel larynx-preserving pharyngectomy technique using a thyroid gland flap after chemoradiotherapy.

This case report describes larynx-preserving pharyngectomy after chemoradiotherapy using a thyroid gland flap. A thyroid gland flap has good blood supply and reconstruction can be done in the same surgical field. The thyroid gland flap has potential as a novel appropriate flap for use in head and neck surgery.

Phyllodes tumor metastasis to the tonsil with synchronous undifferentiated carcinoma.

INTRODUCTION: Breast tumor metastasis to the tonsil is extremely rare. PRESENTATION OF CASE: A 54-year-old woman underwent resection of a breast malignant phyllodes tumor and later presented with metastasis to the lung and the left tonsil. She underwent left lower lobectomy and resection of the left tonsillar tumor. She subsequently developed undifferentiated carcinoma of the right tonsil. She underwent resection of the right tonsillar tumor and chemotherapy was started. The central venous catheter became infected with methicillin-resistant Staphylococcus aureus. Finally, she died. DISCUSSION: To the best of our knowledge, this is the first reported case of a phyllodes tumor metastasizing to the tonsil. Furthermore, morphological and immunohistochemical study revealed that the right tonsillar tumor was irrelevant to the phyllodes tumors. CONCLUSION: We report a case of phyllodes tumor metastasis to the left tonsil which developed undifferentiated carcinoma in the other side of tonsil later. Breast tumor metastasis to the tonsil is rare but it should be considered as a possible diagnosis.

Endolymphatic hydrops revealed by magnetic resonance imaging in patients with acute low-tone sensorineural hearing loss.

OBJECTIVE: Acute low-tone sensorineural hearing loss (ALHL) has been reported to be associated with endolymphatic hydrops (EHs). However, evaluation of the size of the endolymphatic space has not been reported. We attempted to visualize EH in ALHL using magnetic resonance imaging (MRI). STUDY DESIGN: Prospective diagnostic study. SETTING: University hospital. METHODS: We evaluated 25 ears of 25 unilateral ALHL patients. Three-tesla MRI was obtained 24 hours after intratympanic injection of gadolinium (Gd) (n = 5) or 4 hours after intravenous injection of Gd (n = 20). A radiologist blinded to the patients' clinical data classified the degree of EH in the vestibule and cochlea into 3 groups: none, mild, and significant. RESULTS: On the affected sides, cochlear EH was recognized in 23 ears (92%) and was classified as significant EH (n = 15) or mild EH (n = 8); vestibular EH was detected in 22 ears (88%), classified as significant EH (n = 16) or mild EH (n = 6). Cochlear EH was more frequently observed in the affected ear than in the contralateral ear (90% versus 40%, p < 0.05). CONCLUSION: In ALHL, EH was observed not only in the cochlea but also in the vestibule as in Ménière's disease.

Peak width in multifrequency tympanometry and endolymphatic hydrops revealed by magnetic resonance imaging.

OBJECTIVE: To investigate the relationship between the peak width of the characteristic "M"-shaped peak of 2 kHz conductance tympanometry and the degree of endolymphatic hydrops in magnetic resonance imaging (MRI) after intratympanic or intravenous gadolinium administration. STUDY DESIGN: Prospective study. SETTING: An academic university hospital. PATIENTS: One hundred twenty-eight ears in which multifrequency tympanometry was performed and endolymphatic space size was evaluated by MRI. Forty-five patients were examined bilaterally and 38 patients were examined unilaterally. INTERVENTIONS: Endolymphatic space size was evaluated by MRI after intratympanic or intravenous gadolinium injection. MAIN OUTCOME MEASURES: Endolymphatic space size was classified into three groups: none, mild, and significant in the cochlea and in the vestibule. The relationship between the degree of endolymphatic hydrops and peak width of 2 kHz conductance tympanometry was investigated. RESULTS: The peak width in 94 ears in which significant endolymphatic hydrops was observed on MRI in the cochlea and/or the vestibule was 178.8 ± 102.7 daPa. The peak width in 21 ears in which mild but not significant endolymphatic hydrops was observed on MRI in the cochlea and/or the vestibule was 126.0 ± 77.1 daPa. The peak width in 13 ears with no endolymphatic hydrops in the cochlea and vestibule was 107.1 ± 84.1 daPa. The peak width in ears with significant endolymphatic hydrops was larger than that observed in ears with no endolymphatic hydrops. However, the peak width was not significantly different between cases of mild and absent endolymphatic hydrops. CONCLUSION: Large peak width in multifrequency tympanometry was associated with significant endolymphatic hydrops.

Comparison of computed tomography and magnetic resonance imaging for evaluation of cholesteatoma with labyrinthine fistulae.

OBJECTIVES/HYPOTHESIS: To compare preoperative computed tomography (CT) and magnetic resonance imaging (MRI) findings to the clinical features in patients with middle ear cholesteatoma with labyrinthine fistulae. STUDY DESIGN: Retrospective case series. METHODS: Twenty-eight patients who underwent surgery for middle ear cholesteatoma with one or more labyrinthine fistulae confirmed by CT were studied. Preoperative imaging analysis was performed using three-dimensional fluid-attenuated inversion recovery (3D-FLAIR) MRI. Fistula size measured by CT and the signal intensity ratio (SIR) between the affected lesion in the inner ear and the cerebellum measured by 3D-FLAIR MRI were evaluated with respect to the clinical features and surgical findings. RESULTS: The SIR was not correlated with fistula size. The hearing threshold determined by preoperative bone conduction correlated with the SIR, especially in patients with acute sensorineural hearing loss, but it did not correlate with fistula size. Patients with fistula symptoms had a significantly higher SIR than those without symptoms, and similar findings were observed in patients with an active infection. Patients with a larger fistula or higher SIR tended to have a deeper fistula and a more adhesive fistula matrix at operation. CONCLUSIONS: SIR was more strongly correlated than CT findings to the clinical status of patients with labyrinthine fistulae caused by cholesteatoma. Adhesion of the cholesteatoma matrix to the membranous labyrinth correlated with the SIR; thus, SIR may accurately reflect the extent of inner ear dysfunction with a labyrinthine fistula.

Contrast enhancement of the inner ear in magnetic resonance images taken at 10 minutes or 4 hours after intravenous gadolinium injection.

CONCLUSION: Contrast enhancement of the inner ear by three-dimensional fluid-attenuated inversion recovery magnetic resonance imaging (3D-FLAIR MRI) taken 4 h after intravenous gadolinium (Gd) injection was better than when taken at 10 min. Using heavily T2-weighted 3D-FLAIR MRI, visualization of endolymphatic hydrops (EH) was possible in the vestibule and the cochlea after a standard intravenous dose of Gd. OBJECTIVES: To define a suitable time point for imaging Gd uptake in the inner ear acquired with heavily T2-weighted 3D-FLAIR MRI after standard intravenous Gd administration. METHODS: Using a 3 Tesla MRI unit, heavily T2-weighted 3D-FLAIR MRI images were taken twice at approximately 10 min (conventional timing) and 4 h after intravenous gadodiamide (0.1 mmol/kg) injection in 10 patients with inner ear diseases including Ménière's disease. RESULTS: The 4 h delay increased Gd enhancement of the 3D-FLAIR MRI images of the perilymphatic space in both symptomatic and asymptomatic ears. The increase in Gd enhancement was greater in symptomatic than in asymptomatic ears. Using this heavily T2-weighted 3D-FLAIR technique, EH was observed in both the cochlea and vestibule in images taken 4 h after the intravenous Gd injection.