SARS-CoV-2 infection increases airway bleeding risk in patients after tracheostomies.

BACKGROUND: Airway bleeding events are a rare incident in SARS-CoV-2-infected patients after tracheostomies. We aimed to explore the correlation between airway bleeding and SARS-CoV-2 infection and evaluate the consistency of SARS-CoV-2 RNA test results in the upper and lower airway samples from patients after tracheostomies. METHODS: Forty-four patients after temporary or permanent tracheostomy were divided into a positive group (29 patients) and a negative group (15 patients) based on the SARS-CoV-2 RNA test results of their oropharyngeal swabs. The oropharyngeal and tracheal swabs of the positive group were re-collected for SARS-CoV-2 RNA detection. Demographic and clinical characteristics and airway bleeding events were recorded for all enrolled patients. RESULTS: Airway bleeding was reported in eleven patients of the positive group (11/29), with seven displaying bloody sputum or hemoptysis, and four featuring massive sputum crust formation in the trachea that resulted in dyspnea, and only one patient in the negative group (1/15), with a significant difference in the airway bleeding rate (37.9% vs. 6.7%, p < 0.05). The SARS-CoV-2 RNA test results showed a statistical difference in cycle threshold (Ct) values between oropharyngeal swabs and tracheal swabs (p < 0.05). CONCLUSIONS: After tracheostomies, patients are more susceptible to airway bleeding if they are infected with SARS-CoV-2. The findings signify that in addition to droplet transmission through tracheostoma, SARS-CoV-2 may infect the oropharynx by airborne and close contact transmission, and that given the higher viral load and longer infection time in the trachea, tracheal swabs are more reliable for SARS-CoV-2 detection in these patients.

Identification and validation of N6-methyladenosine (m6A)-related lncRNAs signature for predicting the prognosis of laryngeal carcinoma, especially for smoking patients.

Laryngeal cancer (LC), a highly fatal tumor in the head and neck region, has been the focus of research in recent years. The study of LC has primarily focused on the role of long non-coding RNAs (lncRNAs) in regulating gene expression, as they have emerged as pivotal factors in this biological process. Additionally, a reversible RNA modification called N6-methyladenosine (m6A) has been observed to have a significant impact on gene expression as well. The purpose of this research is to investigate the impact of m6A-related lncRNAs on the prognosis of laryngeal squamous cell carcinoma (LSCC). Specifically, this investigation analyzed the m6A-related regulators' patterns of expression and mutation, encompassing a total of 15 regulators. Drawing upon the expression levels of prognostic m6A-regulated lncRNAs, two distinct lncRNA clusters were identified. Further analysis revealed differentially expressed lncRNAs between these clusters. In addition to studying the expression of lncRNAs, the researchers also examined the distribution of clinical characteristics and the tumor microenvironment (TME) in relation to the identified lncRNA clusters. This provided valuable insights into potential associations between lncRNA expression patterns and the clinical features of LSCC. Through the establishment of a risk model associated with lncRNAs, we were able to further investigate their clinical features, prognosis, and immune status. Additionally, we conducted a separate analysis of LINC00528, a lncRNA associated with smoking, examining its expression, overall survival time, correlated mRNAs, and conducting enrichment of Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG), as well as determining the sensitivity of related drugs. RT-qPCR results also indicated an increase in LINC00528 expression among smoking LSCC patients. The findings suggest that a high expression level of LINC00528 in LSCC patients may lead to a more favorable prognosis, providing new insights for the management and treatment of LSCC patients, particularly those with high expression of LINC00528. Overall, this research sheds light on the prognostic impact of m6A-regulated lncRNAs in LSCC. The implications of these findings for the advancement of innovative therapeutic approaches for LSCC patients are noteworthy.

Effect of transoral laser microsurgery vs open partial laryngectomy on the prognosis of patients with early laryngeal carcinoma: propensity score-based analysis.

OBJECTIVE: To evaluate the effect of surgical procedures (transoral laser microsurgery (TLM) and open partial laryngectomy (OPL)) on the prognosis of patients with early laryngeal cancer. METHODS: A total of 760 patients diagnosed with early laryngeal cancer (T1-2N0M0) and treated with TLM (n = 416) or OPL (n = 344) between 2004 and 2015 were abstracted from the SEER database. Propensity score matching (PSM) and stabilized inverse probability of treatment weighting (SIPTW) were performed to obtain comparable cohorts. The survival rates were estimated by the Kaplan-Meier method, and compared using the log-rank test. Univariate and multivariate Cox regression analyses with a false discovery rate (FDR) correction were applied to contrast the association between two surgical approaches and overall survival (OS) and disease-specific survival (DSS). RESULTS: The 5-year OS for the TLM group was 79.5% versus 77.7% for the OPL group (P = 0.619). Similar results were revealed for the comparison of 5-year DSS rates (91.1% versus 91.5%, P = 0.891). After PSM and SIPTW balance the confounding factors, no significant difference was observed in the OS and DSS of patients treated with TLM compared to patients treated with OPL. The consistent results were still yielded (all P > 0.05), when stratified by gender, age, year of diagnosis, residence, household income, tumor site, T stage, differentiation, and adjuvant therapy. CONCLUSION: This study provides strong evidence that there is no significant difference in the prognosis of early laryngeal carcinoma between the treatment of TLM and OPL, which may be helpful to guide the clinical decision-making of these patients.

A dynamic prognostic nomogram to predict the benefit from surgical treatment modality for patients with laryngeal squamous cell carcinoma.

BACKGROUND: Although nonsurgical treatment strategy is increasingly adopted in patients with locoregionally advanced laryngeal squamous cell carcinoma (LSCC), survival disparities were reported between surgical treatment modality and organ preservation protocols, highlighting the great importance for accurate patients' selection. METHOD: This secondary analysis used data from the Surveillance, Epidemiology, and End Results database between 2010 and 2015 with follow-up data up to 2018. We developed and validated a dynamic prognostic nomogram for overall survival (OS) in 4237 patients with LSCC and subgroup of 2087 patients with locoregionally advanced laryngeal squamous cell carcinoma (LALSCC). Based on the total risk score derived from the dynamic nomogram, two well-matched risk groups (i.e., low- and high-risk) were created via X-tile software and 1-to-1 propensity score matching (PSM); surgical treatment modality was compared with nonsurgical one in each risk group. RESULTS: A more accurate and convenient dynamic prognostic nomogram based on age, marital status, T category, N category, M category, tumor size, and tumor differentiation was developed and validated, of which the predictive performance was superior to that of TNM staging system. For high-risk LALSCC selected by the dynamic nomogram, after 1-to-1 PSM, significantly improved OS was observed in patients with receiving surgical treatment compared to those receipt of nonsurgical management (restricted mean survival time at 36-month: 26.6 vs 22.7, p < 0.001; restricted mean survival time at 60-month: 36.7 vs 31.0, p = 0.003). CONCLUSION: We establish and validate a more accurate and convenient dynamic prognostic nomogram for patients with LSCC, which may predict the benefit from surgical treatment modality for patients with high-risk LALSCC.

Systemic Analysis of RNA Alternative Splicing Signals Related to the Prognosis for Head and Neck Squamous Cell Carcinoma.

Alternative splicing (AS) is an important mechanism that is responsible for the production of protein diversity. An increasing body of evidence has suggested that out-of-control AS is closely related to the genesis and development of cancer. Systematic analysis of genome-wide AS in head and neck squamous cell carcinoma (HNSCC) has not yet been carried out, and consideration of this topic remains at the preliminary stage and requires further investigation. In this study, systemic bioinformatic analysis was carried out on the genome-wide AS events of 555 clinical HNSCC samples from the TCGA database. Firstly, we statistically analyzed the distributions of seven AS event types in HNSCC samples. Then, through univariate survival analysis, we observed the relationship between AS and the prognosis of the disease and found that 437 intersections of AS events were significantly related to overall survival. Among them, 335 cross-genes showed a high degree of consistency in the genes associated with overall survival and recurrence. The overall survival was significantly related to AS events. Besides, the frequency of overall survival-related ES events was evidently reduced, while the AP and the AT events were increased. In addition, AT events accounted for the largest proportion. Further, multiple regression model analysis proved that AS could become a new classification method for HNSCC, and KEGG enrichment analysis proved that most genes and proteins interacting with AS events had different biological functions and were associated with a variety of diseases. Finally, through the selection of characteristic HNSCC genes and the construction of a prognostic model, seven cross-genes related to survival and recurrence were screened out, and these characteristic genes were verified by multivariate survival model analysis so as to classify the prognosis at different splicing times and gene expression levels. These results have laid a solid foundation for our further research and play a decisive role in showing the correlation of AS with the prognosis of HNSCC.

Defining the regulatory role of programmed cell death 4 in laryngeal squamous cell carcinoma.

Programmed cell death 4 (PDCD4) is decreased in many different kinds of malignant tumors. EMT endows tumor cells invasive and metastatic properties. However, few studies have determined the role of PDCD4 in the regulation of EMT in the context of laryngeal carcinoma. We examined the relationship between PDCD4 and EMT-associated proteins E-cadherin and N-cadherin using laryngeal carcinoma tissues. Gene manipulation was used to define the regulatory capacity of PDCD4. We report that PDCD4 and E-cadherin/N-cadherin expression were significantly changed in the carcinoma tissues, and their expression was associated with pathological grade, metastatic state, and clinical stage. The suppression of PDCD4 (and consequently, E-cadherin) was concomitant with increased proliferation and G2-phase arrest, decreased apoptosis, and increased cell invasion. PDCD4 upregulation reversed the above-mentioned results. In nude mice, PDCD4 knockdown increased tumor growth and pathological features, confirming the tumorigenic role of PDCD4. Finally, PDCD4 silencing was associated with dysregulation of the carcinogenic Wnt-ß-catenin and the STAT3-miR-21 signaling pathways. This study revealed a dynamic regulatory relationship between PDCD4 and critical factors for EMT, establishing a broad, functional role for PDCD4 in laryngeal carcinoma, which may be propagated by the STAT3-miR-21 pathway. These findings provide new information on an EMT-associated target that may lead to a novel therapy.

[The reliable treatment choice of nasopharyngeal angiofibroma and causes of operative bleeding].

OBJECTIVE: To introduce the efficacy of three surgical options for juvenile nasopharyngeal angiofibroma (JNA) resection, and causes of operative bleeding. METHOD: Retrospective analysis of 36 JNAs,three surgical options were used to resect the tumor. There were 15 cases of Class I tumors , using endoscopic nasal cavity approach. Eighteen cases of class II tumors, via extended Caldwell-Luk incision, using the transantral-infratemporal fosse-nasal cavity combined approach for tumor resection. Three cases of class III tumors, the combined intracranial and extra-cranial approach was used to resect the tumor. Meanwhile, report six typical cases for reference. RESULT: Fifteen (15/36) cases of class I tumors, 14 cases were completely resected for the first time without recurrence, 1 recurrence case was re-resected using the same approach. Eighteen (18/36) cases of class II tumors, 13 cases were completely resected for the first time without recurrence, 5 recurrence cases were re-resected totally. Three (3/36) cases of class III were not completely removed, and underwent about 40 Gy radiotherapy with good effects. CONCLUSION: Using these three surgical options can effectively remove different types of JNA. When necessary, the intracranial residue can use radiotherapy. Under direct vision to separate the tumor, and effective hemostasis play crucial roles for complete removal of the tumor.

Detection of Alternatively Spliced or Processed RNAs in Cancer Using Oligonucleotide Microarray.

Deregulation of gene expression plays a pivotal role in tumorigenesis, so the ability to detect RNA alterations is of great value in cancer diagnosis and management. DNA microarrays have been used to measure changes in mRNA or microRNA level, but less often the change of RNA isoforms. Here we appraise the utilization of microarray in detecting alternatively processed RNAs, which have alternative splice forms, retained introns, or altered 3' untranslated regions. We cover the methodology and focus on cancer studies. Recent development in parallel or deep sequencing used in transcriptome analysis is also discussed.

Nasopharyngeal angiofibroma: a concise classification system and appropriate treatment options.

OBJECTIVES: To describe a clear and simplified classification system for juvenile nasopharyngeal angiofibroma (JNA), and to describe suitable management options. STUDY DESIGN: Retrospective medical record review. METHODS: The clinical and imaging materials of 51 cases of JNA diagnosed at our hospital between 1981 and 2011 were collected and studied. Based on our experiences, we prefer to divide JNAs into three types. Type I includes JNAs fundamentally localized to the nasal cavity, paranasal sinus, nasopharynx, or pterygopalatine fossa. Type II is a JNA extending into the infratemporal fossa, cheek region, or orbital cavity, with anterior and/or minimal middle cranial fossa extension but intact dura mater. Type III is a calabash-like massive tumor lobe in the middle cranial fossa. The management and prognosis for the three types of JNA were compared and evaluated. RESULTS: Among cases of type I JNA (n=16), the entire mass was removed by the initial operation in 15 cases and by a repeat operation in 1 case. Among cases of type II JNA (n=29), the entire mass was removed by the first operation in 24 cases and by repeat operation in 5 cases. In cases of type III JNA (n=6), the huge calabash-like lobe in the middle cranial fossa could not be completely excised; 4 cases underwent radiotherapy and 2 cases were lost to follow-up. CONCLUSIONS: 1) The transnasal cavity approach with endoscopic guidance is suitable for type I JNA resection. 2) The transantral-infratemporal fossa-nasal cavity combined approach is reliable for resection of a type II JNA, which extends into the deep anterior cranial fossa and/or minimally into the middle cranial fossa, with intact dura mater. 3) The complete removal of a type III JNA is difficult, even through a combined extracranial and intracranial approach. Radiotherapy is useful for treating the residual intracranial tumor. The successful or failed experiences of 6 typical cases prove that this revised classification system is reasonable and reliable.

[Study and analysis on the hemorrhage of pterygoid venous plexus in large nasopharyngeal angiofibroma resection].

OBJECTIVE: To our knowledge, study of the intraoperative profuse bleeding of pterygoid venous plexus (PVP) in large nasopharyngeal angiofibroma resection has not yet been reported. Attention should be paid to this topic in clinical practice. METHOD: From 1981 to 2009, 44 cases of JNAs were treated in our hospital. Twenty-six of 44 cases were large nasopharyngeal angiofibromas according to the Fisch classification system(Fisch type III 16, type IV 10). The amount of intraoperative blood loss in these 26 cases varied from 200 ml to 5200 ml. Factors influencing intraoperative bleeding of 26 large nasopharyngeal angiofibroma resections were analyzed retrospectively. The intra-operative observations and imaging data of three typical cases were hereby studied. RESULT: After embolization of the tumor-supplying branches of the external carotid artery(ECA), both the intraoperative observations and imaging data demonstrated that the pterygoid venous plexus (PVP) played a crucial role in intraoperative hemorrhage. CONCLUSION: PVP in the infratemporal fossa communicates with craniofacial veins. There is no valve between these veins. Once PVP is seriously damaged, venous blood of all craniofacial veins will flow out profusely. In the first operation, the intact PVP in the fatty pad generally can be identified and separated from the tumor by delicate surgical managements. If an unsuccessful operation due to serious hemorrhage had been done previously, then scar tissue might tightly adhere with PVP, tumor and the pterygoid muscles, and separation of the tumor from PVP without bleeding is more difficult. Appropriate surgical approach and correct hemostatic procedure of every bleeding point should be done carefully under direct vision. Using finger or instrument for quick blind dissection should be prohibited.

[Perioperative managements of huge lobulated nasopharyngeal angiofibromas with intracranial extensions].

OBJECTIVE: To summarize our experience of successful and failed management in 8 huge lobulated nasopharyngeal angiofibromas with intracranial extensions, and introduce some key points of perioperative treatments. METHOD: Eight male case with an average age of 18 years, were all lobes extending into middle and/or anterior cranial fossa, in which 5 cases revealed blood supply from the internal carotid arteries and 3 cases were reoperated because of recurrence. Preoperatively, the tumor were evaluated by CT, CTA, MRI and/or MRA, and super selective embolization of the feeding arteries were crucial procedures. The combined craniofacial approaches were used to excise these tumors. RESULT: Five cases were removed completely, and 3 cases were removed partly in which 2 were due to serious bleeding caused by lack of DSA technique at that time and 1 were due to neglecting the tumor lobe in the sphenoid sinus of the other side. CONCLUSION: Reasonable perioperative management are very important for control of intra-operative blood loss, complete remodeling of the tumor and avoiding complication.

[Pathological features and clinical managements of nasopharyngeal angiofibroma].

OBJECTIVE: To study the pathological features of nasopharyngeal angiofibroma (NA) and the principles for clinical managements. METHODS: Thirty-five patients with NAs were treated in First Affiliated Hospital of Fujian Medical University from Oct. 1981 to May 2007. The pathological changes, sites of origin, causes of intraoperative bleeding and the experiences of managements were retrospectively analysed. Using Fish stage: 6 cases were in stage I, 8 cases were in stage II, 17 cases were in stage III, 4 cases were stage IV. Two cases via endoscopic surgery, 2 cases via palatal approach, 19 cases via midfacial degloving approach, 9 cases via lateral rhinotomy approach, 3 cases via craniofacial combined approach. RESULTS: In nasal cavity and paranasal sinus, the tumor was covered by squamous or columnar epithelium. The tumor extensions such as in pterygopalatine fossa and infratemporal fossa were covered by fibrous pseudocapsule. All cases of this series originated in the lateral wall of posterior portion of the nasal cavity. Fifteen of thirty-five cases confidentially originated near sphenopalatine foramen. Large and thick vessels in the pedicle region were the exact sites of serious intraoperative bleeding. Thirty-one cases were totally removed. Four cases were subtotal resected. Visual loss revealed in 6 cases, 4 cases visual acuity improved postoperatively. Three cases revealed postoperative dry eye due to surgical involvement of the sphenopalatine ganglion. CONCLUSIONS: nasopharyngeal angiofibroma is covered by epithelium or pseudo-capsule, it does not infiltrate the surrounding tissue. Dissecting along the surface of tumor might decrease bleeding and facilitate removal of tumor. An ideal surgical management should be done according to actually size and image examination, to the greatest extent keeping normal facial appearance. Attention should be paid to the complications such as visual loss and dry eye.

[Expression and its significance of STAT3, STAT5, Survivin and Ki67 in nasal NK/T cell lymphoma].

OBJECTIVE: To study the expression and its significance of STAT3, STAT5, Survivin and Ki67 in the Epstein-Barr virus associated nasal NK/T cell lymphoma. METHOD: The expression of STAT3, STAT5, Survivin and Ki67 were detected with immunohistochemistry in 25 cases of nasal NK/T cell lymphoma, and their relationship was analyzed. Nasal cavity tissues from 20 cases of chronic sinusitis were as the control group. RESULT: The positive rates of STAT3 in the lymphoma and in control group were 56% and 10%, respectively (P <0.01). The positive rates of STAT5 in the lymphoma and in control group were 68% and 15%, respectively (P <0.01). The positive rates of Survivin and Ki67 of nasal NK/T cell-lymphoma were 68% and 72%, respectively; but it in the control group was 0% and 20%, respectively, P <0.01. The expression of STAT3 was positively related to that of Survivin and Ki67(r =0. 428, P <0. 05 and r =0. 704, P <0. 01); The expression of STAT5 was not related to that of Survivin and Ki67. CONCLUSION: The pathway of STAT may play a role in the development of nasal NK/T cell lymphoma. STAT maybe take part in the formation of nasal NK/T cell lymphoma through induction of Survivin and Ki67.

[Primary functional analysis of CK13 gene 5' flanking region].

OBJECTIVE: To investigate the mechanism of cytokeratin 13 (CK13) gene expression control and the effects of different motifs of CK13 gene 5' flanking region on its transcriptional activity. METHODS: The molecular clone technique and reporter gene analysis were used to assay the effects of different motifs of 513 bp of CK13 gene 5' flanking region on its transcriptional activity. The pCAT enhancer vectors with different motifs of CK13 gene 5' flanking region were constructed and transferred to HeLa cells with the help of lipofectin. The instant CAT expression of different clones was detected and the effects of different motifs of the CK13 gene 5' flanking region on its transcriptional activity were evaluated. RESULTS: 119 bp from -nt.325 to -nt.207 upstream of the first ATG of CK13 gene 5' flanking region included a silent element. 113 bp region from -nt.206 to -nt.94 included an enhanced element. CONCLUSION: 513 bp of CK13 gene 5' flanking region includes a silent element and an enhanced element. Further locating these cis elements and detecting the related trans reaction factors may unveil some important clues to the details of the mechanisms for the CK13 gene expression and tissue-specific expression.

[Identification of AP1 cis-element and transcriptional effect on cytokeratin 13 gene expression].

OBJECTIVE: To identify the type of CTGAATCA from -nt.199 to -nt.192 of the cytokeratin 13(CK13) gene 5' flanking region and determine its transcriptional effect on CK13 gene expression. METHODS: The CAT systems were used to assess the effects of different motifs of CK13 gene 5' flanking region on transcription. The clones of pCAT-enhancer with the total length, -nt.207 to +nt.63 and the same length of -nt.207 to +nt.63, but the T, G of -nt.198, -nt.197 being changed to A, T of the CK13 gene 5' flanking region, were constructed and transferred to HeLa cells with the help of lipofectin. Then work was done to detect the instant CAT expression of different clones and evaluate the effects of CTGAATCA of the 5' flanking region on CK13 gene expression. The type of the cis-element of CTGAATCA was identified with electrophoretic mobility shift assay (EMSA) and competition-EMSA. RESULTS: CTGAATCA in the CK13 gene 5' flanking region is an AP1 cis-element by EMSA and competition-EMSA, it promotes CK13 gene expression. CONCLUSION: CTGAATCA from -nt.199 to nt.192 of the CK13 gene 5' flanking region is an AP1 reaction element, not a cAMP reaction element. It promotes transcriptional activity of CK13 gene 5' flanking region.