GFP-labeled Schwann cell-like cells derived from hair follicle epidermal neural crest stem cells promote the acellular nerve allografts to repair facial nerve defects in rats.

BACKGROUND: Acellular nerve allografts (ANAs) have been successfully applied to bridge facial nerve defects, and transplantation of stem cells may enhance the regenerative results. Up to now, application of hair follicle epidermal neural crest stem cell-derived Schwann cell-like cells (EPI-NCSC-SCLCs) combined with ANAs for bridging facial nerve defects has not been reported. METHODS: The effect of ANAs laden with green fluorescent protein (GFP)-labeled EPI-NCSC-SCLCs (ANA + cells) on bridging rat facial nerve trunk defects (5-mm-long) was detected by functional and morphological examination, as compared with autografts and ANAs, respectively. RESULTS: (1) EPI-NCSC-SCLCs had good compatibility with ANAs in vitro. (2) In the ANA + cells group, the GFP signals were observed by in vivo imaging system for small animals within 8 weeks, and GFP-labeled EPI-NCSC-SCLCs were detected in the tissue slices at 16 weeks postoperatively. (3) The facial symmetry at rest after surgery in the ANA + cells group was better than that in the ANA group (p < 0.05), and similar to that in the autograft group (p > 0.05). The initial recovery time of vibrissal and eyelid movement in the ANA group was 2 weeks later than that in the other two groups. (4) The myelinated fibers, myelin sheath thickness and diameter of the axons of the buccal branches in the ANA group were significantly worse than those in the other two groups (P < 0.05), and the results in the ANA + cells group were similar to those in the autograft group (p > 0.05). CONCLUSIONS: EPI-NCSC-SCLCs could promote functional and morphological recovery of rat facial nerve defects, and GFP labeling could track the transplanted EPI-NCSC-SCLCs in vivo for a certain period of time. These may provide a novel choice for clinical treatment of peripheral nerve defects.

Exosomes from Hair Follicle Epidermal Neural Crest Stem Cells Promote Acellular Nerve Allografts to Bridge Rat Facial Nerve Defects.

Previous studies showed that acellular nerve allografts (ANAs) have been successfully utilized in repairing peripheral nerve defects, and exosomes produced by stem cells are useful in supporting axon regrowth after peripheral nerve injury. In this study, exosomes from hair follicle epidermal neural crest stem cells (EPI-NCSCs-Exos) combined with ANAs were used to bridge facial nerve defects. EPI-NCSCs-Exos were isolated by ultracentrifuge, and were identified. After coculture, EPI-NCSCs-Exos were internalized into dorsal root ganglions (DRGs) and schwann cells (SCs) in vitro, respectively. EPI-NCSCs-Exos elongate the length of axons and dendrites of DRGs, and accelerated the proliferation and migration of SCs, and increased neurotrophic factor expression of SCs as well. The next step was to assign 24 Sprague Dawley male rats randomly and equally into three groups: the autograft group, the ANA group, and the ANA + EPI-NCSCs-Exos group. Each rat manufactured a 5-mm gap of facial nerve defect and immediately bridged by the corresponding transplants, respectively. After surgery, behavioral changes and electrophysiological testing of each rat were observed and assessed. At 90 days postoperatively, the retrogradely fluorescent tracer-labeled neurons were successfully observed on the injured side in the three groups. Morphological changes of facial nerve regeneration were evaluated by transmission electron microscopy and semithin toluidine blue staining. The results showed that nerve fiber density, nerve fiber diameter, and myelin sheath thickness in the ANA group were significantly worse than those in the other two groups (P < 0.05). No significant difference in nerve fiber density and myelin sheath thickness was observed between the autograft group and the ANA + EPI-NCSCs-Exos group (P = 0.14; P = 0.23). Our data indicated that EPI-NCSCs-Exos facilitate ANAs to bridge facial nerve defects and have the potential to replace autograft therapy in clinic.

Repairing whole facial nerve defects with xenogeneic acellular nerve grafts in rhesus monkeys.

Acellular nerve allografts conducted via chemical extraction have achieved satisfactory results in bridging whole facial nerve defects clinically, both in terms of branching a single trunk and in connecting multiple branches of an extratemporal segment. However, in the clinical treatment of facial nerve defects, allogeneic donors are limited. In this experiment, we exposed the left trunk and multiple branches of the extratemporal segment in six rhesus monkeys and dissected a gap of 25 mm to construct a monkey model of a whole left nerve defect. Six monkeys were randomly assigned to an autograft group or a xenogeneic acellular nerve graft group. In the autograft group, the 25-mm whole facial nerve defect was immediately bridged using an autogenous ipsilateral great auricular nerve, and in the xenogeneic acellular nerve graft group, this was done using a xenogeneic acellular nerve graft with trunk-branches. Examinations of facial symmetry, nerve-muscle electrophysiology, retrograde transport of labeled neuronal tracers, and morphology of the regenerated nerve and target muscle at 8 months postoperatively showed that the faces of the monkey appeared to be symmetrical in the static state and slightly asymmetrical during facial movement, and that they could actively close their eyelids completely. The degree of recovery from facial paralysis reached House-Brackmann grade II in both groups. Compound muscle action potentials were recorded and orbicularis oris muscles responded to electro-stimuli on the surgical side in each monkey. FluoroGold-labeled neurons could be detected in the facial nuclei on the injured side. Immunohistochemical staining showed abundant neurofilament-200-positive axons and soluble protein-100-positive Schwann cells in the regenerated nerves. A large number of mid-graft myelinated axons were observed via methylene blue staining and a transmission electron microscope. Taken together, our data indicate that xenogeneic acellular nerve grafts from minipigs are safe and effective for repairing whole facial nerve defects in rhesus monkeys, with an effect similar to that of autologous nerve transplantation. Thus, a xenogeneic acellular nerve graft may be a suitable choice for bridging a whole facial nerve defect if no other method is available. The study was approved by the Laboratory Animal Management Committee and the Ethics Review Committee of the Affiliated Wuxi No. 2 People's Hospital of Nanjing Medical University, China (approval No. 2018-D-1) on March 15, 2018.

Nasal Septal Swell Body: A Distinctive Structure in the Nasal Cavity.

OBJECTIVES: The nasal septal swell body (NSB), also known as the nasal septal turbinate, is located in the anterior part of the nasal septum. This study is a narrative review of the existing knowledge on recent developments in NSB. METHODS: A literature search was performed using PubMed, Embase, Web of Science, Ovid, and Cochrane Library databases. Google Scholar was used to access more extensive literature. The inclusion criteria were human studies published in English. The exclusion criteria were non-English language and animal studies. RESULTS: Of the 345 articles that were initially obtained from 5 databases and Google Scholar, 28 were included in this review. There have been many names for NSBs in the past, which still have no unified terminology recognized by professionals. Pathological investigations revealed that NSB contains a certain amount of sinusoidal blood components. Nasal septal swell body is closely related to the internal nasal valve. Imaging studies have found that the size of NSB is associated with nasal diseases, and NSB hypertrophy can cause anatomic obstruction. In recent years, several procedures for NSB have been reported, and preliminary effectiveness has been achieved. However, the long-term outcomes of volume reduction techniques remain unproven. CONCLUSIONS: The NSB is a distinct anatomic structure that may contribute to nasal obstruction and may be reduced surgically with unclear long-term results. Although being investigated for over a century, the unique physiological roles of NSB are not yet fully understood. More evidence is needed to elucidate its physiological effects.

Expression of estrogen and progesterone receptors in angioleiomyoma of the nasal cavity of six patients.

Angioleiomyoma of the nasal cavity is extremely rare. There are only a small number of studies in the literature that demonstrate that the estrogen receptor (ER) and progesterone receptor (PR) are expressed in angioleiomyoma, and the results from these studies are inconsistent. The present study identified 6 patients with nasal angioleiomyoma that were treated between 2004 and 2013. All patients underwent endoscopic surgery and were followed-up for 1-10 years. Resected tumors were investigated for the presence of ER and PR using immunoperoxidase staining. Of the 6 patients, 4 were men and 2 were woman. The mean age of the patients was 60.5 years. The tumors of the 6 patients were identified in the nasal septum, middle turbinate, inferior turbinate, lateral wall of the nasal cavity and nasal vestibule. The clinical manifestations reported by the patients consisted of a painless mass, recurrent epistaxis and nasal obstruction. There were no specific features observed in any of the patients using computed tomography or magnetic resonance imaging. All the patients underwent tumor dissection visualized with a nasal endoscope and recovered without recurrence or malignancy of the tumor post-surgery. Hematoxylin and eosin and immunoperoxidase staining confirmed the diagnosis of angioleiomyoma in all patients. In 5 patients the nuclei of the smooth muscle tumor cells markedly expressed ER and PR. To the best of our knowledge, the present study is the first to demonstrate that ER and PR are clearly expressed in nasal angioleiomyoma. The present study suggests that the sex hormones are possibly associated with the growth of angioleiomyoma.

[Giant tonsillolith in a child].

We describe a case of a 7-year-old child with Down syndrome who presented with loud snoring and cessation of breath during sleep and was found to have a large calculus (20 mm X 12 mm X 12 mm) in her left tonsil by CT scan for which tonsillectomy with adenoidectomy were done. This is one of the youngest reported cases in the literature.

[Analysis of basal cell adenoma of the parotid gland: nine cases report].

OBJECTIVE: To improve recognization of clinical, imaging and pathological characteristics of basal cell adenoma (BCA) of parotid gland. METHOD: We collected and analyzed the data of the clinical manifestation, imaging features, histological and immunohistochemical characteristics of BCA of parotid gland (n = 9). RESULTS: Among 9 patients, 2 male, 7 female, and the average age was 55.2 (from 34 to 66 years). The clinical manifestation showed the painless mass in the parotid region. Seven cases underwent CT and other 2 cases underwent MRI. Imaging showed all tumors were located in the superficial lobe, roughly spherical and non-lobulated in shape, with well-defined boundary. The maximal diameter was less than 30 mm. Seven cases showed slightly high density shadow of soft tissue on plain CT. Compared with the parotid signal intensity, two cases showed hypointensity on T1WI and high signal on T2WI. All lesions showed obvious enhancement on delayed contrast CT or MRI. Cystic changes occurred in two cases. Gross observation: the tumors were solid in section, grey, medium in nature and clear with the surrounding tissues. Cystic changes occurred in the superficial region of superficial lobe of two cases. The capsule of tumor was well circumscribed in 8 cases and focally involved in another case. Microscopy: The parenchyma of BCA was mainly composed of basaloid cells, with myoepithelial cells palisading at the periphery of the epithelial nests. Basal membrane separated the parenchyma from the stromal, the latter lacking the myxochondroid matrix. Immunohistochemistry: basaloid cells were positive for CKpan, CD117 and CKL, while myoepithelial cells were positive for P63, SMA, and calponin. The Ki-67 lablel ing index of tumour cell was 0-4%. All patients performed superficial lobe parotidectomy and tumor dissection, and they recovered well postoperatively without tumor recurrence or malignancy. CONCLUSION: The BCA of parotid gland has distinctive imaging and pathological features with favourable postoperative prognosis.

Regeneration of facial nerve defects with xenogeneic acellular nerve grafts in a rat model.

BACKGROUND: Because of ease of harvest and low immunogenicity, xenogeneic acellular nerve graft (XANG) may be an alternative to autologous nerve to repair facial nerve defects. METHODS: Facial nerve defects of Wistar rats were repaired by XANG, and nerve gap regeneration was investigated by electrophysiological test, horseradish peroxidase (HRP) retrograde tracing and histomorphometric analysis, as compared to autograft. RESULTS: Twenty weeks after the grafting, electrophysiology showed that whisker pad muscles responded to the electrical stimuli given at the site proximal to the transplantation in 2 groups. Some HRP-labeled facial motorneurons were located on the facial nucleus of the operated side, and an abundance of myelinated axons were found at the middle of the grafts and obvious motor endplates in the target muscles in 2 groups, although they were inferior to the contralateral side in numbers. CONCLUSION: XANG represents an alternative approach for the reconstruction of peripheral facial nerve defects.

[The clinical value of sentinel lymph node detection in laryngeal and hypopharyngeal carcinoma patients with clinically negative neck by methylene blue method and radiolabeled tracer method].

OBJECTIVE: To investigate the clinical value of sentinel lymph node (SLN) detection in laryngeal and hypopharyngeal carcinoma patients with clinically negative neck (cN0) by methylene blue method, radiolabeled tracer method and combination of these two methods. METHOD: Thirty-three patients with cN0 laryngeal carcinoma and six patients with cN0 hypopharyngeal carcinoma underwent SLN detection using both of methylene blue and radiolabeled tracer method. All these patients were accepted received the injection of radioactive isotope 99 Tc(m)-sulfur colloid (SC) and methylene blue into the carcinoma before surgery, then all these patients underwent intraopertive lymphatic mapping with a handheld gamma-detecting probe and blue-dyed SLN. After the mapping of SLN, selected neck dissections and tumor resections were peformed. The results of SLN detection by radiolabeled tracer, dye and combination of both methods were compared. RESULT: The detection rate of SLN by radiolabeled tracer, methylene blue and combined method were 89.7%, 79.5%, 92.3% respectively. The number of detected SLN was significantly different between radiolabeled tracer method and combined method, and also between methylene blue method and combined method. The detection rate of methylene blue and radiolabeled tracer method were significantly different from combined method (P < 0.05). Nine patients were found to have lymph node metastasis by final pathological examination. The accuracy and negative rate of SLN detection of the combined method were 97.2% and 11.1%. CONCLUSION: The combined method using radiolabeled tracer and methylene blue can improve the detection rate and accuracy of sentinel lymph node detection. Furthermore, sentinel lymph node detection can accurately represent the cervical lymph node status in cN0 laryngeal and hypopharyngeal carcinoma.

[Wide chondrosarcoma of the larynx in female: a case report].

A case of a 67-years-old female with well-differentiated chondrosarcoma of the left lamina of the thyroid cartilage, cricoid cartilage and arytenoid cartilage is reported, in which a total arytenoidectomy and partial resection of the left thyroid cartilage and cricoid cartilage were performed. The postoperative course has been successful except for the existence of a tracheal stoma and slight hoarseness. There has been no evidence of recurrence or metastasis in 6 years of follow-up.

[Study on treatment of 26 cases with nasal basal cell carcinoma].

OBJECTIVE: To discuss the relationship between surgical margin and recurrence of nasal basal cell carcinoma. METHOD: Twenty-six cases of nasal basal cell carcinoma were analyzed. Mohs microsurgical operation was used in 15 cases and conventional operation was used in 11 cases. RESULT: Twenty-six cases of the tumors were resected and the wound defect was repaired concurrently. Two cases with tumor recurrence were subjected secondary resection and then no recurrence occurred. CONCLUSION: Intraoperation frozen section can help guide the surgical margin. Skin tissue was saved and the repair was facilitated, it also help save the skin tissue , facilitate the repair, reduce the recurrence rate but increased the operation cost and time.

[The expression and its potentially clinical significance of EphA2 in nasopharyngeal carcinoma].

OBJECTIVE: To observe the expression of EphA2, and investigate its correlation with the development, progression, invasion, metastasis, angiogenesis and prognosis in nasopharyngeal carcinoma(NPC). METHOD: Immunohistochemical staining was used to determine the expression level of EphA2 protein in 61 cases NPC and 20 cases chronic nasopharyngitis samples. The clinically pathological data and results of follow-up were collected. Microvessel density (MVD) was also measured by immunohistochemical staining method with CD34 in NPC. RESULT: The positive rate of EphA2 protein staining in NPC was 60.66% (37/61), while that in nasopharyngitis samples was 10.0% (2/20). The positive rates of EphA2 protein in NPC were 27.27% (3/11) in stage I, 56.25% (9/16) in stage II, 68.19% (15/22) in stage III, and 83.33% (10/12) in stage IV. The positive expressions of EphA2 in T1 + T2 and T3 + T4 with neck lymph node and distant metastasis were 58.33% (7/12) and 88.89% (16/18) respectively, while those in T1 +T2 and T3 + T4 without metastasis were 31.25% (5/16) and 50.00% (6/12) respectively. The cumulative survival of patients in the EphA2 positive group at 5 years was only 0.324 (12/37), while 0.500 (12/24) in the EphA2 negative group. The positive expression of EphA2 protein was correlated with the clinical stage, the neck lymph node metastasis and distant metastasis, and prognosis of NPC, respectively (P < 0.05). MVD in EphA2 protein positive group (45.32 +/- 4.91) was significantly higher than that in EphA2 protein negative group (28.69 +/- 3.99, P < 0.01). CONCLUSION: EphA2 may play an important role in the development and progression of NPC. It is closely associated with the invasion, metastasis, angiogenesis and prognosis of NPC.

[Modified intranasal endoscopic dacryocystorhinostomy in chronic dacryocystitis].

OBJECTIVE: To evaluate the technique and curative effect of modified intranasal endoscopic dacryocystorhinostomy (EDCR) for chronic dacryocystitis. METHOD: Twenty-two patients (Twenty-three eyes)with chronic dacryocystitis, undergoing modified intranasal EDCR were retrospectively analyzed in this study. RESULT: The follow-up period ranged from six months to ten months. Twenty eyes were cured successfully and two eyes had relieved symptoms. While one case failed. No serious complications were found. The total effective rate was 22/23 (95.7%). CONCLUSION: The modified intranasal EDCR is an effective method to treat chronic dacryocystitis.

[The expression and potentially clinical significance of heparanase in nasopharyngeal carcinoma].

OBJECTIVE: To investigate the expression of heparanase in nasopharyngeal carcinoma and the relationship between the expression of it and clinically pathological features of nasopharyngeal carcinoma. METHOD: The expression of heparanase protein in 70 cases of nasopharyngeal carcinomas and 10 cases of normal nasopharyngeal tissues was detected by immunohistochemical staining. The date of expression combined clinical features, which included clinical stage, cervical lymph node metastasis rate, the rate of metastasis and recurrence, combination of, the 5-year survival rate, and other analysis, was analyzed. RESULT: The positive rate of heparanase protein in cancerous tissues was 52.9% (37/70), while it was 0% in normal nasopharyngeal tissues. The positive rates of heparanase protein in patients were 30.0% (6/20) in stage I, 45.80% (11/24) in stage II, 70.6% (12/17) in stage III, 88.9% (8/9) in stage IV respectively. Heparanase positive tumors were associated with a higher incidence of lymph node metastasis (67.4%, 31/46) than heparanase negative ones (25.0%, 6/24). The rate of distant metastasis and regional recurrence in the heparanase positive group was 48.6% (18/37), but only 15.2% (5/ 33) in the heparanase negative group. The cumulative survival of patients in the heparanase negative group at 5 years was 78.8% (26/33), but only 24.3% (9/37) in the heparanase positive group. The clinical stage of disease, lymph node metastasis, the rate of distant metastasis and regional recurrence of nasopharyngeal carcinoma were correlated with positive expression of heparanase protein. CONCLUSION: The expression of HPA was associated with invasion and metastasis and prognosis of nasopharyngeal cancer, and it may be a new target for the anti-treatment of nasopharyngeal cancer. (P < 0.01), and heparanase expression level inversely correlated with the patient survival (P < 0.01). CONCLUSION: Heparanase may play important roles in the invasive infiltration, metastasis, and prognosis in nasopharyngeal carcinoma, clearly indicating that heparanase is a possible target for anticancer drug development.