I-scan combined with laryngovideostroboscopy for predicting malignancy in vocal fold leukoplakia.

OBJECTIVE: This study evaluated vocal fold leukoplakia using i-scan combined with laryngovideostroboscopy for risk assessment prediction. METHODS: A total of 141 patients with 218 lesions were enrolled in this study. Morphological characteristics of leukoplakia, assessment of the vascular pattern using i-scan, and vocal fold vibratory function were analyzed. RESULTS: The number of patients with no, mild, moderate, severe dysplasia, and invasive carcinoma were 68, 40, 17, 46 and 47, respectively. The sensitivity of morphological characteristic, vascular pattern, vibratory function and predictive model were 77.4%, 72%, 69.9%, and 82.8%, respectively. Receiver operating characteristic curve analysis of morphological characteristic, vascular pattern, vibratory function and predictive model were 0.771, 0.824, 0.769, and 0.923, respectively. The results of logistic regression analysis showed that rough morphological types, perpendicular vascular pattern, severe decrease and absence of mucosal waves increased the risk of malignancy (OR = 5.531, 4.973, and 16.992, respectively; P < 0.001). CONCLUSIONS: I-scan combined with laryngovideostroboscopy can improve the differential diagnosis of low-risk and high-risk vocal fold leukoplakia.

The Role of Cytokines in Modulating Vocal Fold Fibrosis: A Contemporary Review.

OBJECTIVES: Vocal fold (VF) scarring and laryngeal stenosis are a significant clinical challenge. Excessive scar formation causes low voice quality or even life-threatening obstructions. Cytokines are thought to modulate multiple steps of the establishment of VF fibrosis, but there is no systematic report regarding their role in modulating VF fibrosis. This review aims to investigate the role of cytokines in modulating vocal fold fibrosis. STUDY DESIGN: Literature review. METHODS: This review searched for all relevant peer publications in English for the period 2009 to 2019 in the PubMed database using search terms: "laryngeal stenosis," "vocal fold scarring," and "cytokines." A thorough investigation of the methods and results of the reviewed studies was performed. RESULTS: Comprehensive research in various studies, including analyses of prostaglandin E2 (PGE2), granulocyte-macrophage colony-stimulating factor (GM-CSF), hepatocyte growth factor (HGF), basic fibroblast growth factor (bFGF), transforming growth factor-ß3 (TGF-ß3), and interleukin-10 (IL-10), supports cytokine therapy for VF scarring and laryngeal stenosis to some extent. A few clinical studies on this topic support the conclusion that HGF and bFGF can be selected as effective drugs, and no serious side effects were found. CONCLUSIONS: This review describes the potential of cytokines for modulating the process of VF fibrogenesis, although cytokines are still an unproven treatment method. As no ideal drugs exist, cytokines may be considered the candidate treatment for preventing VF fibrogenesis. Laryngoscope, 131:139-145, 2021.

The Antiproliferative and Antifibrotic Effects of Cisplatin on Primary Human Vocal Fold Fibroblasts.

OBJECTIVE: Vocal fold scarring and laryngeal stenosis are major clinical challenges. Current drugs do not efficiently reduce scarring. We examined the antiproliferative and antifibrotic effects of cisplatin on primary human vocal fold fibroblasts (HVFFs). METHODS: HVFFs were cultured in vitro and identified by immunocytochemistry. The relative viability of HVFFs was analyzed by Cell Counting Kit-8 assays (CCK-8). The fibrogenic phenotype was induced by transforming growth factor-ß1 (TGF-ß1) and reversed by cisplatin as shown by immunocytochemistry. Real-time PCR and Western blotting assessed collagen III and I. Western blotting for Smad2, p-Smad2, Smad-3, p-Smad3 and caspase-3 were performed. RESULTS: CCK-8 results showed that cisplatin decreased the relative viability of HVFFs, and Western blots revealed elevation of the apoptosis-related protein caspase-3 in HVFFs. Cisplatin treatment reduced α-smooth muscle actin staining intensity in the presence of TGF-ß1. Real-time PCR revealed the downregulation of collagen III and I in cisplatin-treated HVFFs. The TGF-ß1-induced increased fibrogenic protein levels were decreased by cisplatin. Reduced levels were detected at late time points. CONCLUSIONS: Cisplatin induces antiproliferative and antifibrotic alterations in HVFFs. Cisplatin may prevent postoperative vocal fold scarring and laryngeal stenosis in patients treated with CO2 laser microsurgery and undergoing delayed wound healing.

The primary growth of laryngeal squamous cell carcinoma cells in vitro is effectively supported by paired cancer-associated fibroblasts alone.

Most primarily cultured laryngeal squamous cell carcinoma cells are difficult to propagate in vitro and have a low survival rate. However, in our previous work to establish a laryngeal squamous cell carcinoma cell line, we found that laryngeal cancer-associated fibroblasts appeared to strongly inhibit the apoptosis of primarily cultured laryngeal squamous cell carcinoma cells in vitro. In this study, we investigated whether paired laryngeal cancer-associated fibroblasts alone can effectively support the growth of primarily cultured laryngeal squamous cell carcinoma cells in vitro. In all, 29 laryngeal squamous cell carcinoma specimens were collected and primarily cultured. The laryngeal squamous cell carcinoma cells were separated from cancer-associated fibroblasts by differential trypsinization and continuously subcultured. Morphological changes of the cultured laryngeal squamous cell carcinoma cells were observed. Immunocytofluorescence was used to authenticate the identity of the cancer-associated fibroblasts and laryngeal squamous cell carcinoma cells. Flow cytometry was used to quantify the proportion of apoptotic cells. Western blot was used to detect the protein levels of caspase-3. Enzyme-linked immunosorbent assay was used to detect the levels of chemokine (C-X-C motif) ligand 12, chemokine (C-X-C motif) ligand 7, hepatocyte growth factor, and fibroblast growth factor 1 in the supernatants of the laryngeal squamous cell carcinoma and control cells. AMD3100 (a chemokine (C-X-C motif) receptor 4 antagonist) and an anti-chemokine (C-X-C motif) ligand 7 antibody were used to block the tumor-supporting capacity of cancer-associated fibroblasts. Significant apoptotic changes were detected in the morphology of laryngeal squamous cell carcinoma cells detached from cancer-associated fibroblasts. The percentage of apoptotic laryngeal squamous cell carcinoma cells and the protein levels of caspase-3 increased gradually in subsequent subcultures. In contrast, no significant differences in the proliferation capacity of laryngeal squamous cell carcinoma cells cocultured with cancer-associated fibroblasts were detected during subculturing. High level of chemokine (C-X-C motif) ligand 12 was detected in the culture supernatant of cancer-associated fibroblasts. The tumor-supporting effect of cancer-associated fibroblasts was significantly inhibited by AMD3100. Our findings demonstrate that the paired laryngeal cancer-associated fibroblasts alone are sufficient to support the primary growth of laryngeal squamous cell carcinoma cells in vitro and that the chemokine (C-X-C motif) ligand 12/chemokine (C-X-C motif) receptor 4 axis is one of the major contributors.

Healing process following laser cordectomy of early glottis carcinoma from endoscopic view.

BACKGROUND AND OBJECTIVE: Transoral laser surgery is useful for treating early glottic carcinoma. Wound areas after carbon dioxide (CO2 ) laser surgery are not covered by flaps, delaying healing. The healing process following laser cordectomy is not well understood. In this study, laryngoscopes were used to monitor the normal healing process after laser cordectomy and to determine if there was residual or recurrent disease. MATERIALS AND METHODS: This prospective case study included 36 patients who underwent CO2 laser surgery for early glottis carcinoma between January 2011 and June 2014 at a university tertiary referral center. Postoperative complications, oncologic results, and mucosal stabilization time were analyzed. Vocal fold stabilization was defined as an absence of gross changes during two consecutive follow-up examinations. RESULTS: The 3-year overall survival rate was 100% and the 3-year local control rate was 94%. Vocal cord stabilization was observed in 29 patients at a mean 88.1 days, with stabilization associated with the type of cordectomy (P < 0.05). During the wound-healing process, two patients experienced local recurrences. One had a persistent white patch for over 4 months and the other presented with a new mass in a non-operated area. Biopsies of both lesions showed recurrent disease. Postoperative complications included granulation in two patients and anterior commissure adhesion in one. CONCLUSION: CO2 laser microsurgery is an effective treatment for early glottis carcinoma. Close attention should be paid to delays in healing process after laser surgery. Interventions should be considered for patients with abnormal laryngoscopic appearance or overly prolonged healing. Lasers Surg. Med. 48:483-489, 2016. © 2016 Wiley Periodicals, Inc.

Cancer-associated fibroblasts in a human HEp-2 established laryngeal xenografted tumor are not derived from cancer cells through epithelial-mesenchymal transition, phenotypically activated but karyotypically normal.

Cancer-associated fibroblasts (CAFs) play a crucial role in cancer progression and even initiation. However, the origins of CAFs in various cancer types remain controversial, and one of the important hypothesized origins is through epithelial-mesenchymal transition (EMT) from cancer cells. In this study, we investigated whether the HEp-2 laryngeal cancer cells are able to generate CAFs via EMT during tumor formation, which is now still unknown. The laryngeal xenografted tumor model was established by inoculating the HEp-2 laryngeal cancer cell line in nude mice. Primary cultured CAFs from the tumor nodules and matched normal fibroblasts (NFs) from the adjacent connective tissues were subcultured, purified, and verified by immunofluorescence. Migration, invasion, and proliferation potentials were compared between the CAFs and NFs. A co-culture of CAFs with HEp-2 cells and a co-injection of CAFs with HEp-2 cells in nude mice were performed to examine the cancer-promoting potential of CAFs to further verify their identity. Karyotypic analyses of the CAFs, NFs, and HEp-2 cells were conducted. A co-culture of NFs with HEp-2 cells was also performed to examine the expression of activated markers of CAFs. A pathological examination confirmed that the laryngeal xenografted tumor model was successfully established, containing abundant CAFs. Immunocytochemical staining verified the purities and identities of the CAFs and NFs. Although the CAFs manifested higher migration, invasion, proliferation, and cancer-promoting capacities compared with the NFs, an analysis of chromosomes revealed that both the CAFs and NFs showed typical normal mouse karyotypes. In addition, the NFs co-cultured with HEp-2 cells did not show induced expressions of activated markers of CAFs. Our findings reveal that the CAFs in the HEp-2 established laryngeal xenografted tumor are not of laryngeal cancer origin but of mouse origin, indicating that the HEp-2 laryngeal cancer cells cannot generate their own CAFs via EMT in this model.

Forehead Flap for Simultaneous Reconstruction After Head and Neck Malignant Tumor Resection.

INTRODUCTION: Oncologic resection of the head and neck malignant tumor often results in complex reconstructive problems that require a reliable flap to restore both the form and function in single-stage operation. The free flap transfer has revolutionized head and neck reconstruction. However, forehead flap pedicled with superficial temporal artery (STA) or supraorbital artery (SOA)/supratrochlear artery (STrA) is a new option for reconstruction of head and neck with its own specific advantages. METHOD: The forehead flap was applied to reconstruct the defects of the posterior hypopharyngeal wall, external perioral skin and buccal mucosal, and nose and periorbital tissues after malignant tumor resection. Nine of the patients were reconstructed with forehead flap pedicled with STA, 7 of whom underwent postoperative radiation. Eight of the patients were pedicled with SOA (and/or STrA), 1 of whom underwent postoperative radiation. RESULTS: We applied this method in a total of 17 male patients. The mean age of patients was 61 years (range, 47-69 years) with a follow-up time of 12 to 180 months. The biggest defect covered by forehead flap pedicled with STA was measured 10.0 × 6.5 cm. The biggest defect covered by forehead flap pedicled with SOA and/or STrA was measured 9.0 × 7.0 cm. In all 17 patients, no vascular crisis happened and no partial or total flap necrosis was observed. All skin graft area and donor site healed well. No patient experienced flap strictures or fistulas after operation or after radiotherapy. No local recurrence developed during the follow-up time. Functions were assessed by specialist as optimal as no limitation of eyes or mouth opening, and no limitation of food swallowing. CONCLUSIONS: The forehead flap provides thin pliable tissue with reliable blood supply and high tolerance to radiotherapy. These advantages make it a valuable option to simultaneously reconstruct the defects after head and neck malignant tumor resection in certain selected patients.

Superficial temporal artery flap: a new option for posterior hypopharyngeal wall reconstruction.

Reconstruction of hypopharynx after tumor resection remains a challenging problem for the head and neck surgeons. A new surgical technique was introduced to reconstruct the hypopharynx after tumor resection using the superficial temporal artery flap. Three patients with T3 carcinoma of the posterior hypopharyngeal wall who underwent surgical resection of the tumor with laryngeal preservation were reported using this method. The larynx functions were preserved successfully in all the patients. In all patients, the nasogastric tubes were removed within 20-50 days (mean 33 days). The tracheal tubes were plugged on days between 14 and 44 (mean 26th day) postoperatively. Then all patients received the postoperative radiotherapy. The tracheal tubes were removed successfully 5-7 months after surgery (mean 6 months) without the evidence of apnea. The speech function was excellent in all patients, which was evaluated according to the voice-related quality of life questionnaire. Although lymph node metastasis was detected in one patient 6 months after surgery, there were no evidences of flap complications in all the three patients. The superficial temporal artery flap is a new option for the posterior pharyngeal wall reconstruction in selected cases with the hypopharynx carcinoma. The larynx functions were preserved successfully.

Mandibular distraction osteogenesis reconstructed the upper airway in a case of Melnick-Needles syndrome.

INTRODUCTION: We report a 16-year-old female with Melnick-Needles syndrome complicated with severe obstructive sleep apnea-hypoxia syndrome. CLINICAL REPORT: An overnight sleep study demonstrated that the patient had severe obstructive sleep apnea with an apnea-hypopnea index of 95/h. The lowest oxygen saturation was only 34%. The distraction osteogenesis technique was used to lengthen her mandible for the reconstruction of her upper airway. RESULT AND DISCUSSION: Postoperative panorex showed that the bilateral body of her mandible had been distracted about 16 mm singulorumly in the end of our distraction. Lateral radiograph revealed that the diameter of oral pharynx widen nearly 5 mm. The patient recovered after the upper airway reconstruction.

[Clinical analysis of 10 cases of olfactory neuroblastoma].

OBJECTIVE: To review the diagnosis, treatment and prognosis of olfactory neuroblastoma. METHODS: Clinical data were retrospectively reviewed in 10 cases of olfactory neuroblastoma admitted from 1998 to 2002, including data of transmission electron microscopic (TEM) observation in 4 cases. According to Kadish's classification, 2 cases were in stage A, 4 in stage B and 4 in stage C. Three patients were treated with surgery alone, 7 with combined surgery and radiation. RESULT: Among 10 cases, the overall 5-year survival rate was 60 %(6/10); 3 patients died from local recurrence, 1 lost follow-up. TEM demonstrated granules in the cytoplasm of 3 patients. CONCLUSION: The combined surgery and radiation can achieve excellent local control. Transmission electron microscope is important for its diagnosis.

The efficacy of melilotus extract in the management of postoperative ecchymosis and edema after simultaneous rhinoplasty and blepharoplasty.

BACKGROUND: Melilotus extract has been used as a folk medicine in inflammation-related therapy. This study was undertaken to determine the potential benefits of this herbal extract in treating postoperative ecchymosis and edema after simultaneous rhinoplasty and blepharoplasty. METHODS: Augmentation rhinoplasty and double-eyelid blepharoplasty were performed in one procedure by the same surgeon on 46 patients. RESULTS: Postoperative edema and ecchymosis were evaluated independently by two investigators checking the upper eyelid, lower eyelid, and paranasal area at postoperative days 1, 4, and 7. Our data showed that the patients receiving melilotus extract (n = 16) had lower scores not only in upper-eyelid and lower-eyelid ecchymosis, but also in paranasal ecchymosis at postoperative day 7 when compared with those in the control group. No significant difference in postoperative ecchymosis was detected in the group receiving dexamethasone treatment (n = 16) when compared with the control group (n = 14) at postoperative days 1, 4, and 7. The mean scores of upper-eyelid, lower-eyelid, and paranasal edema at postoperative days 1 and 4 were significantly lower in the group receiving dexamethasone treatment (n = 16) than that in the control group. Although the scores of eyelid edema reduction in the patients receiving melilotus extract showed no difference when compared with the control group, paranasal edema was reduced much more at postoperative days 4 and 7 when compared with the control group. CONCLUSION: We suggest the appropriate use of melilotus extract based on its beneficial effects on the postoperative results after simultaneous rhinoplasty and blepharoplasty.

[Management of sinonasal inverted papilloma: endoscopic approach and lateral rhinotomy].

OBJECTIVE: To compare endoscopic approach with lateral rhinotomy for treatment of the sinonasal inverted papilloma in terms of advantage, indications and limitations of the procedures. METHODS: Eighty-six cases with inverted papilloma were reviewed retrospectively, among which 23 cases underwent transnasal endoscopic procedures including 10 combined with Caldwell-Luc intervention, and 63 cases underwent lateral rhinotomy. The follow-up period ranged from 11 - 36 m (mean 23 m). The data were processed statistically by SPSS 10.0 software. RESULT: Both procedures permitted removal of most sinonasal inverted papilloma. The endoscopic surgery provided an excellent visualization, and preserved a vital anatomic structure and left no facial scar. Lateral rhinotomy was associated with postoperative facial scar or deformity. The recurrence rate in lateral rhinotomy group was 9.5% and in endoscopic approach was 13% (P >0.05). CONCLUSION: Endoscopic approach is favored for the treatment of non-massively extending sinonasal inverted papilloma because of an acceptable recurrence and a better cosmetic results.

Eosinophil infiltration and activation in nasal polyposis.

Conclusions. Infiltration and activation of eosinophils is a characteristic of nasal polyposis. Allergic reaction is a risk factor for the accumulation of eosinophils in this disease. T-cell-derived interleukin 5 (IL-5) and autosecretion of IL-5 from activated eosinophils may be causative reasons for the extension and persistence of eosinophil inflammation. Objectives. To investigate whether eosinophils were accumulated and activated in nasal polyposis, and the roles of IL-5, eotaxin, and T cells in this process. Materials and methods. A retrospective study was conducted on 17 tissue samples from patients with nasal polyposis with allergy and 26 cases of non-allergic polyposis. Immunohistochemical staining by specific antibodies was carried out using the alkaline phosphatase anti-alkaline phosphatase method and the avidin-biotin complex technique.Results. The number of EG1-positive cells (pan eosinophil marker) was similar to the number of EG2-positive cells (activated eosinophil marker) in all tissue samples, although EG1- and EG2-positive cells were richer in allergic patients than those in non-allergic patients. Both EG1- and EG2-positive cells were correlated with CD3-positive cells (pan T cell marker) and IL-5-producing cells in allergic or non-allergic polyposis. A large proportion of IL-5 producing cells were eosinophils. Eotaxin protein was detected in all tissue samples and dominantly located in epithelial cells. Eotaxin expression between allergic and non-allergic subjects was not significantly different.

Characteristics of nasal inverted papilloma and its malignant transformation: a study of cell proliferation and programmed cell death.

BACKGROUND: In nasal inverted papilloma (IP) and its malignant transformation process, abnormal cell growth has been regarded as a altered proliferation of the epithelial cells, while the role of programmed cell death in this process remains to be determined. METHODS: An apoptotic index was calculated, and the proliferation ability by using Ki67 as a marker was evaluated in the study subjects, which consisted of 15 cases of nasal chronic sinusitis, 23 cases of nasal IP, 9 cases of nasal IP with dysplasia, 8 cases of nasal IP with squamous cell carcinoma (SCC), and 21 cases of primary SCC. In addition, expressions of apoptosis-related molecules p53, Bcl-2, Bax, and Fas were investigated also. RESULTS: The result showed that the Ki67 index increased in IP and IP with dysplasia. The Ki67 value was further enhanced in SCC within IP and primary SCC. The apoptotic index was low in IP with dysplasia, IP with SCC, and primary SCC. Concerning the expression of apoptosis-related proteins, p53 protein accumulated in IP with dysplasia, IP with SCC, and primary SCC. The p53 overexpression was directly correlated with the Ki67 index in IP and primary SCC diseases. No remarkable differences were found regarding expression of Bcl-2 and Fas productions among these diseases, and Bax protein was decreased only in primary SCC. CONCLUSION: We concluded that inhibition of apoptosis could be an early sign of IP undergoing malignant transformation. A high proliferative rate was a characteristic of IP-associated malignant diseases.

p57(kip2) expression is related to carcinogenesis and tumor progression in laryngeal tissues.

CONCLUSIONS: Reduction of p57(kip2) expression is related to carcinogenesis in laryngeal mucosa. Assessment of p57(kip2) expression may be a promising candidate as a progressive biomarker in laryngeal squamous cell carcinoma (SCC). OBJECTIVE: To investigate the expression and significance of p57(kip2) in a dysplasia/carcinoma sequence of laryngeal mucosa. MATERIAL AND METHODS: A retrospective study was performed involving 10 cases of normal mucosa, 31 of laryngeal leukoplakia and 109 of laryngeal SCC. p57(kip2) and p53 protein expression was detected in the tissue samples of these patients using immunohistochemistry. RESULTS: p57(kip2) expression was decreased in leukoplakia with moderate or severe dysplasia, and further decreased in SCC. No significant difference was found between normal mucosa and leukoplakia with no or mild dysplasia. In patients with laryngeal SCC, advanced tumor size and clinical stage and the recurrence of disease were associated with reduced p57(kip2) expression. The occurrence of lymph node metastasis was also related to negative expression of p57(kip2). The overall 5-year disease-free survival rate of patients with laryngeal SCC was significantly higher in the p57(kip2)-positive than in the p57(kip2)-negative group.

Immunohistochemical analysis of P57(kip2), p53 and hsp60 expressions in premalignant and malignant oral tissues.

To investigate the expressions and significances of P57(kip2), p53 and hsp60 proteins in the dysplasia-carcinoma sequence of oral mucosa. A retrospective study was performed in 10 cases of normal oral mucosa, 79 cases of leukoplakia and 67 cases of squamous cell carcinoma (SCC). The expressions of P57(kip2), p53 and hsp60 proteins were detected in the tissue samples of these populations using immunohistochemical method. P57(kip2) expression was decreased in oral leukoplakia with moderate or severe dysplasia, and further decreased in oral SCC. Negative expression of P57(kip2) was significantly associated with advanced tumor size, the occurrence of lymph node metastasis and the advanced clinical stage in oral SCC. The overall 5-year survival rate in the P57(kip2) positive group was significantly higher than that in the P57(kip2) negative group. P57(kip2) expression was decreased in oral leukoplakia with moderate or severe dysplasia, and further decreased in oral SCC. It was a remarkable progressive and prognostic biomarker in oral SCC.