Deciphering head and neck cancer microenvironment: Single-cell and spatial transcriptomics reveals human papillomavirus-associated differences.

Human papillomavirus (HPV) is a major causative factor of head and neck squamous cell carcinoma (HNSCC), and the incidence of HPV- associated HNSCC is increasing. The role of tumor microenvironment in viral infection and metastasis needs to be explored further. We studied the molecular characteristics of primary tumors (PTs) and lymph node metastatic tumors (LNMTs) by stratifying them based on their HPV status. Eight samples for single-cell RNA profiling and six samples for spatial transcriptomics (ST), composed of matched primary tumors (PT) and lymph node metastases (LNMT), were collected from both HPV- negative (HPV- ) and HPV-positive (HPV+ ) patients. Using the 10x Genomics Visium platform, integrative analyses with single-cell RNA sequencing were performed. Intracellular and intercellular alterations were analyzed, and the findings were confirmed using experimental validation and publicly available data set. The HPV+ tissues were composed of a substantial amount of lymphoid cells regardless of the presence or absence of metastasis, whereas the HPV- tissue exhibited remarkable changes in the number of macrophages and plasma cells, particularly in the LNMT. From both single-cell RNA and ST data set, we discovered a central gene, pyruvate kinase muscle isoform 1/2 (PKM2), which is closely associated with the stemness of cancer stem cell-like populations in LNMT of HPV- tissue. The consistent expression was observed in HPV- HNSCC cell line and the knockdown of PKM2 weakened spheroid formation ability. Furthermore, we found an ectopic lymphoid structure morphology and clinical effects of the structure in ST slide of the HPV+ patients and verified their presence in tumor tissue using immunohistochemistry. Finally, the ephrin-A (EPHA2) pathway was detected as important signals in angiogenesis for HPV- patients from single-cell RNA and ST profiles, and knockdown of EPHA2 declined the cell migration. Our study described the distinct cellular composition and molecular alterations in primary and metastatic sites in HNSCC patients based on their HPV status. These results provide insights into HNSCC biology in the context of HPV infection and its potential clinical implications.

Effect of monopolar diathermy power settings on postoperative pain, wound healing, and tissue damage after tonsillectomy: a randomized clinical trial.

This study aimed to assess the impact of varying monopolar diathermy power settings on postoperative pain, hemorrhage, and wound healing following tonsillectomy. A single-center, prospective, randomized, double-blinded, controlled clinical study was conducted. During bilateral tonsillectomy procedures, one tonsil received low-power settings (15 W, cutting/blend) while the other tonsil received high-power settings (35 W, cutting/blend). Postoperative pain scores (0-10) and wound healing scores (0-3) were evaluated immediately after surgery and at 1, 2, and 4 weeks postoperatively using the visual analog scale. Additionally, histological analysis was performed on electrically resected tonsil tissues to assess tissue damage in the tonsil bed. The allocation of high and low power settings to each side was randomized. Results showed that 1 week after the surgery, the high-power group experienced significantly higher pain scores (mean ± standard deviation: 4.84 ± 2.21) compared to the low-power group (3.56 ± 2.24, p = 0.049). Moreover, the high-power side exhibited slower wound healing during the initial 1-2 weeks postoperatively, as indicated by lower wound scores at 2 weeks (high-power: 1.96 ± 0.64; low-power: 2.43 ± 0.59, p = 0.008). Furthermore, histological analysis revealed significantly deeper tissue degradation on the high-power side compared to the low-power side (p < 0.001), with mean depths of 565.2 ± 291.0 µm and 156.0 ± 36.8 µm, respectively. In conclusion, these findings suggest that when employing monopolar diathermy in tonsillectomy, lower power settings can lead to improved outcomes in terms of postoperative pain, wound healing, and tissue damage.Trial registration: CRIS identifier: KCT0005670 (cris.nih.go.kr, registration date: 11/12/2020).

Liquid plasma as a treatment for cutaneous wound healing through regulation of redox metabolism.

The skin functions as the outermost protective barrier to the internal organs and major vessels; thus, delayed regeneration from acute injury could induce serious clinical complications. For rapid recovery of skin wounds, promoting re-epithelialization of the epidermis at the initial stage of injury is essential, wherein epithelial keratinocytes act as leading cells via migration. This study applied plasma technology, which has been known to enable wound healing in the medical field. Through in vitro and in vivo experiments, the study elucidated the effect and molecular mechanism of the liquid plasma (LP) manufactured by our microwave plasma system, which was found to improve the applicability of existing gas-type plasma on skin cell migration for re-epithelialization. LP treatment promoted the cytoskeletal transformation of keratinocytes and migration owing to changes in the expression of integrin-dependent focal adhesion molecules and matrix metalloproteinases (MMPs). This study also identified the role of increased levels of intracellular reactive oxygen species (ROS) as a driving force for cell migration activation, which was regulated by changes in NADPH oxidases and mitochondrial membrane potential. In an in vivo experiment using a murine dorsal full-thickness acute skin wound model, LP treatment helped improve the re-epithelialization rate, reaffirming the activation of the underlying intracellular ROS-dependent integrin-dependent signaling molecules. These findings indicate that LP could be a valuable wound management material that can improve the regeneration potential of the skin via the activation of migration-related molecular signaling within the epithelial cell itself with plasma-driven oxidative eustress.

Liquid-type plasma-controlled in situ crosslinking of silk-alginate injectable gel displayed better bioactivities and mechanical properties.

Silk is a promising biomaterial for injectable hydrogel, but its long-gelation time and cytotoxic crosslinking methods are the main obstacles for clinical application. Here, we purpose a new in situ crosslinking technique of silk-alginate (S-A) injectable hydrogel using liquid-type non-thermal atmospheric plasma (LTP) in vocal fold (VF) wound healing. We confirmed that LTP induces the secondary structure of silk in a dose-dependent manner, resulting in improved mechanical properties. Significantly increased crosslinking of silk was observed with reduced gelation time. Moreover, controlled release of nitrate, an LTP effectors, from LTP-treated S-A hydrogel was detected over 7 days. In vitro experiments regarding biocompatibility showed activation of fibroblasts beyond the non-cytotoxicity of LTP-treated S-A hydrogels. An in vivo animal model of VF injury was established in New Zealand White rabbits. Full-thickness injury was created on the VF followed by hydrogel injection. In histologic analyses, LTP-treated S-A hydrogels significantly reduced a scar formation and promoted favorable wound healing. Functional analysis using videokymography showed eventual viscoelastic recovery. The LTP not only changes the mechanical structures of a hydrogel, but also has sustained biochemical effects on the damaged tissue due to controlled release of LTP effectors, and that LTP-treated S-A hydrogel can be used to enhance wound healing after VF injury.

Preoperative perforator localization in anterolateral thigh free flap using acoustic doppler and computed tomography angiography.

Objectives: Our aim in this study was to investigate if we could predict perforator localization during ALTF elevation, using information from acoustic Doppler (AD) and computed tomography angiography (CTA). Methods: Prospective observational data were collected from H&N cancer patients who received reconstruction with ALTF in Ajou University Hospital Cancer Center from June to December, 2021. Total of 21 cases were included in the analysis. Lower extremity angio-CT scans were used to determine the course and depth of the perforator before surgery. During intraoperative design of the ALTF, the possible location of the perforator was identified by AD. After flap elevation, the distance between the actual and Doppler-identified location of the perforator was measured. Results: The average distance from the actual location to the Doppler-identified location was 1.29 ± 1.26 cm. Among 21 cases, almost all perforators (20 cases) were identified in a circle with a radius equivalent to the depth of the perforator. Perforator depth measured by CTA showed a significant positive correlation with the distance from the actual to Doppler-identified location, regardless of skin thickness or body mass index (BMI). Conclusions: A circle with a radius equivalent to the CTA-assessed depth of the perforator successfully predicted the location of the perforator in almost all cases. Depth of the perforator measured by CTA combined with Doppler-identified location can help safely locate the perforator during ALTF harvesting.Level of Evidence: 4.

Taste Dysfunction after Tonsillectomy: A Meta-analysis.

OBJECTIVES: We aimed to provide information on the effects of tonsillectomy, in particular, its effects on taste dysfunction; also known as dysgeusia, and provide direction for future research in this area. METHODS: We independently searched five databases (PubMed, SCOPUS, Embase, the Web of Science, and the Cochrane database) for related studies published as recently as December 2019. From the included studies, we reviewed the findings on tonsillectomy and taste disturbance through the use of a questionnaire, chemogustometry, and electrogustometry. We analyzed other conditions that were measured in these studies that included; postoperative pain intensity and other morbidities (eg, postoperative bleeding, hemostasis, tongue numbness and zinc, copper, and iron hemoglobin levels measured). RESULTS: Postoperative taste disturbance based on questionnaire (VAS) scores at 14 days (SMD = -0.31 [-0.50, -0.12], P value = .0012) tended to decrease significantly in the post-tonsillectomy group compared to the pre-tonsillectomy group. Patients showed significantly decreased taste sensation after tonsillectomy than in the preoperative test. Postoperative electrogusmetry at 7 days (SMD = 0.60 [0.27, 0.94], P = .0004) tended to increase significantly in the post-tonsillectomy group compared to the pre-tonsillectomy group. CONCLUSION: Tonsillectomy is a commonly performed surgery, and taste disturbance occurs frequently as a post operative effect. Taste dysfunction is important to patients with regard to their quality of life and the overall effects of taste on their day-to-day life. This study can be a useful meta-analysis of dysgeusia in tonsillectomy.

[Biphenotypic Acute Leukemia with BCR-ABL mRNA Transcript b3a2 Type: A Case Report with Review of the Literature.].

Biphenotypic acute leukemia (BAL) is a subtype of leukemia of ambiguous lineage in the World Health Organization classification system. About one third of the cases have the Philadelphia chromosome, and some cases are associated with other structural abnormalities involving 11q23. BAL is known to have a poor prognosis in both children and adults. According to the previously reported BAL cases with positive BCR-ABL fusion gene, most of the BCR-ABL mRNA transcript type was e1a2. So, we describe here a 30-year-old adult BAL case with the karyotype 46,XY,t(9;22)(q34;q11.2) resulting in a very rare b3a2 type of BCR-ABL mRNA transcript.