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.

Endoscopic dacryocystorhinostomy with an otologic T-type ventilation tube in repeated revision cases.

BACKGROUND: To compare the frequency of appearance of complications, anatomical success and functional success after conventional endoscopic dacryocystorhinostomies (EN-DCRs) or EN-DCR with otologic T-Type ventilation tube combined with silicone tube intubation in repeated revision cases. METHODS: Twenty-two patients who had epiphora and recurrent dacryocystitis after at least a previous failed revision DCR as well as 22 patients receiving conventional EN-DCR only were enrolled in the study between January 2008 and December 2011. Operations were performed by using an otologic T-tube combined with silicone tube intubation. Oral antibiotics, nasal steroids, oral antihistamines, and antibiotic eyedrops were given to all cases. The ventilation tubes were removed 6 to 20 weeks after surgery. RESULTS: Of 22 cases, all cases achieved anatomical success, 19 cases were symptom free, and 3 cases had decreased continuation in complications with a functional success rate of 81.8%. The overall success rates were significantly higher than those in patients undertaking conventional EN-DCR only (P < 0.01). CONCLUSION: The revision endoscopic DCR has a high rate of failure. The usage of a T-type ventilation tube can significantly improve the success rate of surgery. TRIAL REGISTRATION NUMBER: ChiCTR-INR-17012160, retrospectively registered on July 27th, 2017.

Oleuropein enhances radiation sensitivity of nasopharyngeal carcinoma by downregulating PDRG1 through HIF1α-repressed microRNA-519d.

BACKGROUND: Oleuropein (OL) is a well-known anti-oxidative agent and is shown to reduce the hypoxia-inducible factor 1 α (HIF1α) protein expression after radiation. The current study investigated the effects of OL on radiation response in nasopharyngeal carcinoma (NPC). METHODS: Colony formation assay was performed to compare the radiation response in vitro. Xenograft mouse model was used to study the OL effects on radiation in vivo. Chromatin immunoprecipitation and luciferase reporter assays were performed to identify the relations among HIF1α, miR-519d and PDRG1. Stable HIF1α or PDRG1 overexpression, and miR-519d downregulation were performed to test the radiation response both in vitro and in vivo. RESULTS: OL strongly enhanced radiosensitivity of NPC cells both in vitro and in vivo. Chromatin immunoprecipitation and luciferase reporter assays suggested miR-519d was a direct target of HIF1α, and PDRG1 was a direct target of miR-519d. Overexpression of HIF1α or PDRG1, and downregulation of miR-519d abolished the radiation sensitizing effects of OL. CONCLUSION: Our study hereby demonstrates OL is a radiation sensitizing agent in NPC both in vivo and in vitro. OL treatment reduces the activity of HIF1α-miR-519d-PDRG1 pathway, which is essential to the radiosensitizing effects of OL.

Effect of curcumin on nasal symptoms and airflow in patients with perennial allergic rhinitis.

BACKGROUND: Allergic rhinitis (AR) is a common disorder that can significantly affect patient quality of life. Previous studies have found that curcumin had anti-inflammatory and antioxidant effects and clinical benefits in cancer and asthma. OBJECTIVE: To determine the efficacy of curcumin in the treatment of AR and to explore the molecular mechanisms involved. METHODS: In a randomized, double-blind study, 241 patients with AR received either placebo or oral curcumin for 2 months. The therapeutic effects of curcumin were evaluated by nasal symptoms and nasal airflow resistance. In addition, the production of interferon γ, interleukin (IL) 4, IL-10, and tumor necrosis factor α from mononuclear cells and IL-8, soluble intercellular adhesion molecule, polyethylene glycol 2, and leukotriene C4 from polymorphonuclear neutrophils were compared before and after curcumin treatment. RESULTS: Curcumin alleviated nasal symptoms (sneezing and rhinorrhea) and nasal congestion through reduction of nasal airflow resistance. Curcumin was found to exert diverse immunomodulatory effects, including suppression of IL-4, IL-8, and tumor necrosis factor α and increased production of IL-10 and soluble intercellular adhesion molecule. However, curcumin did not affect the release of prostaglandin E2 and leukotriene C4 from polymorphonuclear neutrophils. CONCLUSION: This pilot study provides the first evidence of the capability of curcumin of improving nasal airflow and modulating immune response in patients with AR.

miRNA-24-3p promotes cell proliferation and regulates chemosensitivity in head and neck squamous cell carcinoma by targeting CHD5.

AIM: To investigate the role of miR-24-3p in tumorigenesis and chemosensitivity in head and neck squamous cell carcinoma (HNSCC). METHODS: Growth rate and colony formation assays were performed after transfection with miR-24-3p mimic and inhibitor in cultured SCC-15 cells, followed by a CellTiter-Glo® assay. Western blot and luciferase assays were performed to investigate the direct target of miR-24-3p. Xenograft mouse model was used to evaluate combinatorial effects of miR-24-3p inhibitor and 5-fluorouracil. RESULTS & CONCLUSION: Inhibition of miR-24-3p reduced cell proliferation, colony formation efficiency and reversed chemoresistance in HNSCC cells. CHD5 is the direct target of miR-24-3p which is required for the regulatory role of miR-24-3p in chemoresistance. miR-24-3p may represent a new therapeutic target for the improvement of clinical outcome in HNSCC.

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.

Contributions of T lymphocyte abnormalities to therapeutic outcomes in newly diagnosed patients with immune thrombocytopenia.

T cell abnormalities have been reported to play an important role in pathogenesis of immune thrombocytopenia (ITP) besides specific autoantibodies towards platelet. The aim of this study was to explore the clinical importance of T lymphocyte subsets in adult patients with newly diagnosed ITP before and after first-line treatment. Elderly ITP patients were also studied and we tried to analyze the relationships between these items and therapeutic outcomes. The patients were treated with intravenous immunoglobulin (IVIG) plus corticosteroids and therapeutic responses were evaluated. As a result, compared with the controls, absolute lymphocyte counts in ITP patients decreased significantly before treatment. After treatment, lymphocyte counts restored to control level regardless of their treatment outcomes. In addition, we observed increased IgG and CD19+ cell expression and decreased CD4+/CD8+ cell ratio in both whole ITP group and elderly group before treatment. After treatment, the increased IgG and CD19+ cell expression could be reduced in both respond and non-respond group regardless of patient age, while CD4+/CD8+ cell ratio could not be corrected in non-respond ITP patients. In non-respond ITP patients, increased CD8+ cell expression was noticed and could not be corrected by first-line treatment. Furthermore, even lower NK cell expression was found in non-respond elderly patients after treatment when compared with that in controls. Our findings suggest that ITP patients usually had less numbers of peripheral lymphocytes and patients with higher levels of CD8+ cells or lower levels of CD4+/CD8+ cell ratio were less likely to respond to first-line treatment. Lower levels of NK cells made therapies in elderly ITP patients even more difficult.

Slug mediates nasopharyngeal carcinoma radioresistance via downregulation of PUMA in a p53-dependent and -independent manner.

Slug is involved in the radioresistance and chemoresistance of several types of cancers. In the present study, we first studied the effect of Slug on the radioresistance of nasopharyngeal carcinoma (NPC). We established radioresistant CNE-2 cells (CNE-2-RES) by exposing CNE-2 cells to gradually increasing doses of irradiation (IR). We used lentiviral infection technique to stably knock down Slug and then studied the effects in vitro and in vivo. Western blotting and RT-PCR were applied to detect the protein and mRNA expression in NPC cells or xenograft tumor tissues, respectively. Colony forming assay was applied to detect the cell survival after IR. As a result, CNE-2-RES cells were successfully established, CNE-2-RES cells showed relatively higher expression of Slug, higher expression of p53 and lower expression of PUMA. Following inhibition of Slug, the radiosensitivity of NPC was enhanced both in vitro and in vivo. Slug inversely regulated PUMA and p53 expression in both CNE-2 and CNE-2-RES cells. Animal experiments showed the same trend of protein expression as the in vitro results. In conclusion, our study demonstrated that Slug overexpression in CNE-2-RES cells may result in the radioresistance of cells. Slug mediates CNE-2 radioresistance via downregulation of PUMA in both a p53-dependent and p53-independent manner.

[The comparison of clinical features of 2 cases of intracranial otogenic complications].

Two cases of special intracranial otogenic complications were analyzed in the aspects of clinical characteristics, diagnosis and therapy. We concluded that for patients with huge cholesteatoma which damaged the bone of skull base, or chronic otitis media patients with sharp deterioration in symptoms, accompanied by headache and fever, we should promptly do the enhanced magnetic resonance to avoid the missed diagnosis of intracranial complications.

Knockdown of phosphodiesterase 4D inhibits nasopharyngeal carcinoma proliferation via the epidermal growth factor receptor signaling pathway.

Phosphodiesterase 4D (PDE4D) is a subtype of metallohydrolases, and it has been reported that PDE4D functions as a proliferation promoting factor in certain types of cancer, including head and neck cancer. The present study first investigated the function of PDE4D in nasopharyngeal carcinoma (NPC). Western blot analysis was applied to detect PDE4D expression in NPC samples and cells. A lentiviral infection technique was used to stabilize the knockdown of PDE4D, which was subsequently examined in vitro and in vivo. The results showed that PDE4D was overexpressed in the NPC tissues and cells. Knockdown of PDE4D inhibited the growth of CNE2 and 5-8F, inducing cell cycle arrest in the G0/G1 phase in CNE2. These effects could be reversed by epidermal growth factor (EGF) stimulation. Furthermore, knockdown of PDE4D significantly inhibited the phosphorylation of epidermal growth factor receptor (EGFR) and AKT. The results were further validated in an NPC xenograft in nude mice. In conclusion, this study demonstrated that PDE4D may function as a proliferation promoting factor in NPC, by affecting the EGFR/PI3K/AKT signaling pathway. Therefore, the targeting of PDE4D may be a rational strategy in the treatment of NPC.

[Mastoidectomy in the treatment of secretory otitis media].

OBJECTIVE: To investigate mastoidectomy efficacy in treating secretory otitis media. METHOD: Retrospective analysis of 22 cases (24 ears) with chronic secretory otitis media,20 ears were treated with intact canal wall mastoidectomy combined with facial recess opening,4 ears were treated with opened mastoid surgery,3 ears simultaneously accepted tube insertion. Ventilation tube was pulled out in 6 months. Hearing test was inspected before and after surgery. RESULT: None of the patients had hearing loss, 19 ears had varying degrees of hearing improvement. Seventeen ears were type A tympanometry curve, 7 ears were C-shaped curve. No recurrence of otitis media was observed after 6 - 36 months followed-up. CONCLUSION: Mastoidectomy may improve eustachian tube function and decrease the risk of recurrence of secretory otitis media.

[Clinical efficacy of mouse nerve growth factor in the treatment of sudden deafness].

OBJECTIVE: To study the clinical efficacy of mouse nerve growth factor (NGF) in the treatment of sudden deafness. METHOD: A retrospective analysis was performed on 115 cases of hospitalized patients who were suffered from sudden deafness. Patients were divided into two groups according to treatment medicine. Control group: patients were treated with intravenous vasodilators, energy mixture, steroid pulse therapy, and methylcobalamin neurotrophic therapy. NGF group: intramuscular NGF treatment was added on the basis of conventional therapy mentioned above. Both treatments lasted 14 days, the total efficiency were compared. Patients were further divided into sub-groups according to age, duration and the level of pre-treatment PTA, and the treatment efficiency was further compared. By SPSS 11.0 statistical analysis, a P < 0.05 was considered as statistical significant difference. RESULT: (1) The total efficiency of NGF group was significantly higher than control group. (2) Regard of age, the efficiency of NGF treatment group was significantly higher than control group. (3) For the patients whose duration were less than 7 d, or the PTA < or = 60 dBHL, the efficiency of NGF group were significantly higher. For the patients whose duration were more than 7 d, or the PTA>60 dBHL, the efficiency of NGF therapy was not superior to the traditional treatment. CONCLUSION: NGF can significantly improve the symptom of patients with short duration or low PTA. For this kind of patients, NGF adjuvant therapy should be recommended. For the patients with longer duration and higher level of PTA, NGF therapy is not advocated. NGF treatment should not be in consideration of the age.

[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.

ECRG4 inhibits growth and invasiveness of squamous cell carcinoma of the head and neck in vitro and in vivo.

ECRG4 has been shown to be a candidate tumor suppressor in several tumors, but its role in head and neck cancer remains poorly understood. In the present study, the effect of ECRG4 on head and neck cancer was investigated in vitro and in vivo. pFLAG-CMV-2-ECRG4 was stably transfected into squamous cell carcinoma of the head and neck (SCCHN) M2 cell lines to overexpress the ECRG4 gene. Real-time PCR and western blot analysis were performed to detect gene and protein expression, respectively. An MTT assay and flow cytometric analysis were used to detect the growth of M2 cells. Matrigel™ invasion and scratch assays were applied to observe the invasion and migration of the cells. A tumorigenicity assay was applied to test the tumor growth and cervical lymph node metastasis in vivo. Based on the data, pFLAG-CMV-2-ECRG4 significantly increased the expression of ECRG4 in the M2 cells. The constructed plasmid inhibited cell proliferation and promoted cell cycle arrest and apoptosis in the M2 cells. The growth rate and metastasis of the tumor cells in xenografts were suppressed following the overexpression of ECRG4 in nude mice. These data suggest that ECRG4 plays a significant role in the regulation of growth and metastasis in SCCHN, providing new clues for the diagnosis and therapy of SCCHN.

[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.

[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 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.

[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.