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Objective: To investigate the clinical efficacy of acellular dermis matrix combined with cervical strap muscle (ADM-CSM) as a composite tissue flap for repairing the laryngeal defect after partial laryngectomy. Methods: The medical records of 33 patients with laryngeal cancer who were diagnosed and treated at the First Affiliated Hospital of Sun Yat-sen University from January 2014 to December 2016 were retrospectively reviewed. The patients consisted of 32 males and 1 female with age range from 39 to 76 years. Laryngeal defects were repaired with ADM-CSM in 14 patients (2 for supraglottic laryngeal cancer, 12 for glottic laryngeal cancer) and with CSM fascial flaps in 19 patients (3 for supraglottic laryngeal cancer, 16 for glottic laryngeal cancer). Kaplan-Meier method was used to calculate the 3-year overall survival and local control rate. The functions of voice and swallowing after operation were evaluated by voice handicap index-30 (VHI-30) and MD Anderson dysphagia inventory. Univariate logistic regression analysis, t -test, and Mann-Whitney U test were used to compare the variables between the two groups. Results: The incidence of laryngeal stenosis was 2/14 in ADM-CSM group and 4/19 in CSM group. In the ADM-CSM group, 3-year overall survival and local control rates were 92.9% and 85.7%, respectively. In the CSM group, 3-year overall survival and local control rates were 78.9% and 84.2%, respectively. The time of operation(3 h vs . 4 h, Z =193.5, P <0.05), time of retaining the feeding tube(14 d vs . 17 d, Z =206.0, P <0.05), and length of stay(18.5 d vs . 22.1 d, t =-2.62, P <0.05) in the ADM-CSM group were significantly less than those in the CSM group. The quality of voice in the CSM group was better than that in the ADM-CSM group (66.85±27.65 vs .45.80±23.19, t =2.19, P <0.05), while swallowing function in the ADM-CSM group was better than that in the ACSM group (80.00[60.00, 80.00] vs .60.00[40.00, 80.00], Z =48.0, P <0.05). Conclusion: ADM-CSM is user-friendly control and safe composite tissue flap for repairing the laryngeal defect after partial laryngectomy, with less scar hyperplasia and higher satisfaction of swallowing function after operation.

Objective: To evaluate the feasibility and efficacy of the integrallty submucosal resection of adult-onset laryngeal papilloma by CO 2 laser. Methods: A group of 64 cases (36 males and 28 females, multipe lesions 54 cases and single lesion 10 cases, aged 18-75 years, mean age 43.13 years) with adult-onset laryngeal papilloma encountered in the first affliated hospital of Sun Yatsen university from 2009 to 2015 was retrospectively analyzed. All cases were treated with integrallty submucosal dissection of the tumor by CO 2 laser, and observed the changes of tumor integral scope, inter-operative, operative processes, postoperative voice quality, postoperative scarring, and the tracheotomy conditions, which were analysed and evaluated. Results: A total of 64 patients were followed up from 1 year to 5 years. Preoperative tumor integral scope of these patients averaged of 7.00. A total of 62 cases kept 0 score of the tumor integral scope for at least one year, which lead to a clinical cure rate of 96.9%. The inter-operative averaged of 25.7 months. The total operative processes of these patients were 87 times (mean time 1.36). Four cases resulted in postoperative scarring. However these was a good result in postoperative voice quality with a mean score 4.25. As to the changes of tumor integral scope, all cases got a declining score (mean score 6.72), which resulted in a remission rate of 100%. Conclusion: The integrallty submucosal dissection of adult-onset 1aryngeal papilloma by CO 2 laser was an effective way to reduce the tumor integral scope; lengthen their inter-operative; decrease the operative processes, avoid the occurrence of tracheotomy; and improve the postoperative voice quality. Most of the patients could even be cured ultimately.

DJ-1, a novel mitogen-dependent oncogene, has an important role in the progression of human malignancies, whereas tumor suppressor phosphatase and tensin homolog (PTEN) is known to control a variety of processes associated with cell survival, proliferation and invasion. DJ-1 overexpression was reported to be negatively correlated with PTEN expression in tumor tissues of patients with laryngeal squamous cell carcinoma (LSCC). In the present study, the effect of DJ-1 on PTEN in laryngeal cancer cells was investigated by transfecting DJ-1-specific small interfering (si)RNA into Hep-2 and SNU-899 cells. Cell survival and cell proliferative and invasive capacity were then evaluated. The results showed that siRNA targeting of DJ-1 effectively upregulated PTEN expression, resulting in enhanced cell death as well as decreased proliferation and invasion of Hep-2 and SNU-899 cells. The results of the present study indicated, for the first time, to the best of our knowledge, that DJ-1-induced PTEN downregulation is associated with proliferative and invasive activity of laryngeal cancer cells. The DJ-1 gene may have an important role in the tumorigenesis of LSCC.

CD4+ regulatory T cells (Tregs) mediate immune tolerance in laryngeal squamous cell carcinoma (LSCC). However, Tregs are functionally heterogeneous. Recently, we reported that three distinct Treg subsets (resting Tregs, activated Tregs and cytokine-secreting CD45RA-Foxp3lowCD4+ T cells) vary in the peripheral circulation of patients with head and neck squamous cell carcinoma (HNSCC); however, the potential implication of these Treg subsets in LSCC immunity is unclear. Here, we report that activated Tregs and cytokine‑secreting CD45RA-Foxp3lowCD4+ T cells were increased in LSCC patients compared with healthy donors (HD) (p<0.001, p<0.001), whereas resting Tregs were decreased (p<0.001). Activated Tregs inhibited the proliferation of CD4+CD25- T cells (p<0.001) and secreted lower levels of interleukin-2 (p<0.001), interferon-γ (p<0.001) and tumor necrosis factor-α (p<0.001) compared with the cytokine-secreting CD45RA-Foxp3lowCD4+ T cells. Importantly, activated Treg prevalence was correlated with tumor stage (p=0.001) and nodal status (p=0.007). The prevalence of naïve CD4+ (p<0.001), naïve CD8+ (p=0.002), and Th1 T-cell subsets (p<0.001, p<0.001) was decreased in the LSCC patients. In conclusion, our findings showed that activated Tregs with suppressive activity are a distinct subset of Tregs in LSCC, and correlate with disease progression. Several immune system abnormalities in LSCC patients are represented by expansion of functionally activated Tregs, both in the circulation and tumor microenvironment along with decreased frequencies of naïve T-cell populations and Th1-cell populations.

Background: The clinical efficiency and functional outcomes of supracricoid partial laryngectomy (SCPL) with cricothyroidopexy (CTP) were compared with those of the traditional SCPL with cricohyoidoepiglottopexy (CHEP) in treating laryngeal squamous carcinoma involving anterior vocal commissure (AVC)., Methods: From January 2000 to June 2009, 50 patients diagnosed with early- or intermediate-stage (T1b-T3 classification) glottic cancer involving AVC were treated with SCPL-CHEP or SCPL-CTP. Postoperative complications, local recurrence, survival rate, and speech performance were compared between these 2 surgical procedures., Results: Patients undergoing SCPL-CHEP or SCPL-CTP manifested similar levels of postoperative complications, tumor recurrence, and survival rates. However, the SCPL-CTP group showed significantly lower Voice Handicap Index (VHI) scores, higher maximum phonation time, and improved glottic reconstruction and closure than the SCPL-CHEP group., Conclusion: The SCPL-CTP procedure better preserves postoperative speech performance than the SCPL-CHEP procedure, underscoring the moderate effectiveness of SCPL-CTP as a treatment for laryngeal squamous carcinoma involving AVC., (Copyright © 2012 Wiley Periodicals, Inc.)

Objective: The therapeutic effect of middle frontal horizontal partial laryngectomy (MFHPL) in treating stage T1b squamous cell carcinoma of the glottic larynx involving anterior vocal commissure (AVC) was compared with that of the anterior frontolateral vertical partial laryngectomy (AFVPL). The feasibility and practical significance of MFHPL in clinical application was discussed in the present study., Methods: From January 1996 to January 2010, a total of 65 patients diagnosed with stage T1bN0M0 glottic laryngeal cancer were treated with MFHPL or AFVPL. The postoperative complications, glottic reconstruction, recurrence rate, voice quality and survival rates were evaluated and compared between two treatments., Results: AFVPL and MFHPL were performed in 34 and 31 patients, respectively. Flexible fiberoptic laryngoscopy revealed that in the MFHPL-treated patients the reconstructed glottis was spacious and symmetric. In contrast, AFVPL treatment resulted in irregular glottic area with poor symmetry and tubular glottis. The incidence of postoperative laryngeal stenosis significantly differed between the MFHPL- and AFVPL-treated groups (P = 0.025). No significant difference was detected in the 3- and 5-year overall- or tumor-free survival rates between two treatments. The Voice Handicap Index (VHI) and maximum phonation time (MPT) after surgery were 51.0±12.99 and 12.42±3.44 sec in the AFVPL-treated group; while in the MFHPL-treated patients they were 31.81±7.48 and 7.65±1.98 sec, respectively. Both differences in VHI (P = 0.012) and MPT (P = 0.024) were significant between two treatments., Conclusions: MFHPL was comparable to AFVPL with respect to postoperative complications, recurrence rate and survival rates, but possessed advantages over AFVPL in terms of the incidence of laryngeal stenosis and voice quality. Our study indicated that MFHPL has a potential value in clinical practice of treating stage T1b squamous cell carcinoma of the glottic larynx involving AVC.

Background: DJ-1 can induce the tumor cell proliferation and invasion via down-regulating PTEN in many malignant tumors, and correlated to prognostic significance. However, the tumorigenesis role and clinical significance of DJ-1 in supraglottic squamous cell carcinoma (SSCC) is unclear. We aimed to evaluate the DJ-1 the relationship between DJ-1 and clinicopathological data including patient survival., Methods: The expression of DJ-1 and PTEN in SSCCs (52) and adjacent non-cancerous tissues (42) was assessed by immunohistochemistry (IHC), and the relationship between DJ-1 and clinicopathological data was analyzed., Results: DJ-1 was detected mainly in SSCCs (88.5%) and less frequently in adjacent non-cancerous tissues (21.0%). PTEN expression was detected in 46.2% of SSCCs and in 90.5% of adjacent non-cancerous tissues. DJ-1 expression was linked to nodal status (P = 0.009), a highly significant association of DJ-1 expression with shortened patient overall survival (5-year survival rate 88.0% versus 53.9%; P = 0.007; log rank test) was demonstrated., Conclusions: Our data suggested that DJ-1 over-expression was linked to nodal status, and might be an independent prognostic marker for patients with SSCC.

Objective: To detect the expression of DJ-1 in laryngeal squamous cell carcinoma (LSCC) and to study the relationship between DJ-1 expression and clinical indexes of LSCC., Methods: The expressions of DJ-1 protein in 71 LSCC samples and 9 cases control samples from laryngeal mucosa tissues of non-LSCC patients were detected using streptavidin peroxidase immunohistochemistry staining and the relationships between DJ-1 protein expression and clinicopathologic characteristics were analyzed., Results: (1) The positive expression rate of DJ-1 protein in LSCC was 85.9%(61/71), which was significantly higher than the rate (55.5%, 5/9) in control laryngeal mucosa tissues (P < 0.05). (2) DJ-1 expression was related to tumor recurrence (P < 0.05), but not to sex, age, primary cancer position, T stage, clinical stage, lymph node metastasis and tumor differentiation. Tumor recurrence rate (53.3%) in the patients with higher expression of DJ-1 protein was higher than the rate (26.8%) in the patients with lower expression of DJ-1 protein (χ(2) = 5.164, P < 0.05). (3) With Kaplan-Meier curves and Cox regression analysis, the cumulative 5-year survival rates were correlated with DJ-1 expression levels in laryngeal cancer tissues or cervical lymph node metastasis (all P < 0.05), but not to sex, age, primary cancer position, T stage, clinical stage and tumor differentiation., Conclusions: The expression of DJ-1 protein in LSCC is higher than that in control laryngeal mucous tissues. Overexpression of DJ-1 is associated with poor overall survival in LSCC patients.

DJ-1 is frequently overexpressed in a large variety of solid tumors, but the DJ-1 expression in laryngeal squamous cell cancer and its clinical/prognostic significance is unclear. We aimed to evaluate DJ-1 protein expression in glottic squamous cell carcinoma (GSCC) and to correlate this with clinicopathological data including patient survival. The expression of DJ-1 in GSCCs (60) and adjacent normal tissue (44) was assessed by immunohistochemistry and western blot analysis. In addition, the role of DJ-1 was investigated in tumorigenesis by transfecting DJ1-specific siRNA into laryngeal squamous cell carcinoma (LSCC) Hep-2 cells. Our data showed that positive expression of DJ-1 was found in 85% of GSCCs. In univariate survival analysis of the GSCC cohorts, a highly significant association between DJ-1 expression with shortened patient overall survival (5-year survival rate 92.9%vs 66.6%; P = 0.001; log rank test) was demonstrated. In multivariate analyses, DJ-1, tumor grading, and pT status were significant prognostic parameters for shortened patient overall survival. Furthermore, siRNA targeting DJ-1 can effectively inhibit DJ-1 expression, resulting in enhanced apoptosis and less proliferation of Hep-2 cells. We concluded that DJ-1 overexpression might be a novel independent molecular marker for poor prognosis (shortened overall survival) of patients with GSCC.

Objective: To evaluate the effect of combined use of rapamycin and cisplatin in the chemotherapy of Hep-2 cells in vitro., Methods: The inhibitory effects of rapamycin and cisplatin, used alone or in combination, on the proliferation of Hep-2 cells were measured with MTT assay and median-effect plot analysis. The cell cycle changes after the treatment were analyzed using flow cytometry and Hoechst 33258 immunofluorescence staining., Results: The IC50 of rapamycin and cisplatin for inducing growth arrest of Hep-2 cells was 11.03 nmol/L and 8.81 micromol/L, respectively. Rapamycin alone caused cell cycle arrest of the Hep-2 cells in G1 phase. Rapamycin and cisplatin showed synergistic effects in the chemotherapy of Hep-2 cells (q > 1.15, King's Formula), causing significantly increased apoptosis ratio and growth inhibition rate of Hep-2 cells., Conclusion: Combined use of rapamycin and cisplatin significantly improves the chemotherapeutic effect against Hep-2 cells.

Objective: To assess the effect of small interfering RNA (siRNA)-mediated gene silencing of DJ-1 on the proliferation of human laryngeal carcinoma cell line Hep-2., Methods: Three siRNA sequences specific to DJ-1 gene were synthesized according to GenBank. Human laryngeal carcinoma cell line Hep-2 was cultured and divided into 4 groups: non-specific group (siRNA control) and 3 RNAi groups, transfected with specific DJ-1 siRNA (siRNA1, siRNA2, siRNA3). The mRNA and protein levels of DJ-1 were detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western Blot respectively. Cell apoptosis were analyzed by flow cytometry. The proliferation of Hep-2 cells was assessed by MTT assay., Results: DJ-1 siRNA down-regulated the mRNA and protein levels of DJ-1 in Hep-2 cells. After transfection, the expression of DJ-1 mRNA and protein levels in Hep-2 cells of the DJ-1 siRNA1 group were significantly lower than those of non-specific siRNA control group. MTT assay showed that DJ-1 siRNA1 group inhibited proliferation of Hep-2 cells. Flow cytometry showed that apoptosis rate of the DJ-1 siRNA1 group (15.7%) was significantly higher than that of non-specific siRNA control group (4.5%) or untransfected group (3.5%), t = 4.736, P < 0.01., Conclusions: Specific siRNA targeting DJ-1 can effectively inhibit DJ-1 expression, resulting in the reduced proliferation and the enhanced apoptosis in Hep-2 cells.

Xin Xia,1,* Ying-Ying Zhu,1,* Wen-Wen Diao,1 Xiao-Li Zhu,1 Xiao-Hua Shi,2 Wu-Yi Li,1 Zhi-Qiang Gao,1 Guo-Jun Li,1 Xing-Ming Chen1 1Department of Otolaryngology-Head and Neck Surgery, Peking Union Medical College Hospital, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing, China; 2Department of Pathology, Peking Union Medical College Hospital, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing, China *These authors contributed equally to this work Background: The survival outcomes between supracricoid partial laryngectomy (SCPL) and total laryngectomy (TL) were compared in the groups of matched-pair patients with T3 laryngeal squamous cell carcinoma (LSCC).Methods: Patients with T3 LSCC were matched based on prognostic factors. The Kaplan–Meier curve and the Cox proportional hazards model were used for analysis on survival.Results: A total of 212 patients with T3 LSCC were included after matching (106 underwent SCPL and 106 underwent TL). Multivariable analysis showed no differences in overall survival (hazard risk [HR]=1.15; 95% CI: 0.79–1.67; P=0.47), disease-specific survival (HR=1.11; 95% CI: 0.69–1.80; P=0.66), and recurrence-free survival (HR=1.07; 95% CI: 0.68–1.68; P=0.77) between the SCPL group and TL group.Conclusion: SCPL provides reliable therapeutic outcomes and can be used to avoid a TL surgery in some patients with advanced primary laryngeal cancer. Keywords: survival analysis, matched-pair analysis, laryngeal neoplasms, laryngectomy, squamous cell carcinoma