Objective To establish adult rat auditory epithelial cell culture and try to find precursor cells of auditory hair cells in vitro. Methods With refinement of culture media and techniques, cochlear sensory epithelial cells of adult rat were cultured. Immunocytochemistry and Bromodeoxyuridine (BrdU)labeling were used to detect properties and mitotic status of cultured cells. Results The cultured auditory epithelial cells showed a large, flat epithelial morphotype and expressed F-actin and cytokeratin, a subset of cells generated from auditory epithelium were labeled by calretinin, a specific marker of early hair cell. Conclusion Adult rat auditory epithelium can be induced to generate hair cell-like cells by nature culture, this phenomenon suggests that progenitor cells may exist in rat cochlea and they may give birth to new hair cells. Whether these progenitor cells are tissue specific stem cells is still need more study.

Objective: To explore the sensitization effects of resveratrol on CNE2 nasopharyngeal carcinoma cell line with hypoxia-induced chemotherapy resistance and the potential mechanism. Methods: Human CNE2 nasopharyngeal carcinoma cell line was cultured under hypoxic conditions (37 degrees centigrade, 5% CO(2), 2% O(2)) in vitro. The cultured cells were treated with different concentrations of resveratrol for 48 h. Reversal fold (RF) of reseratrol to chemotherapeutic drugs in CNE2 cells was measured by methyl thiazolyl tetrazolium (MTT) assay. Apoptotic rate of CNE2 cells was observed by flow cytometry. Reverse transcription-polymerase chain reaction (RT-PCR) method and Western blotting were used to investigate the expressions of multidrug resistance gene 1 (mdr1), multidrug resistance-associated protein 1 (MRP1) and hypoxia inducible factor 1alpha (HIF-1alpha) in CNE2 cells. Results: Resveratrol combined with chemotherapeutics produced a synergistic effect. The RF of 200 alphamol/L resveratrol to paclitaxel was 2.58. Combined with paclitaxel, 25, 50, 100 and 200 alphamol/L of resveratrol increased the apoptotic rate of CNE2 cells from (22.14+/-1.09)% to (23.24+/-1.37)%, (27.57+/-2.01)%, and (30.36+/-2.31)%, respectively. Resveratrol could down-regulate the expressions of HIF-1alpha, mdr1 and MRP1 significantly. After being treated with resveratrol at different concentrations separately, the expressions of HIF-1alpha, mdr1 and MRP1 in CNE2 cells decreased significantly as compared with paclitaxel alone or paclitaxel plus verapamil (P<0.01). Conclusion: Resveratrol can enhance the sensitivity of CNE2 cells to chemotherapeutic drugs under hypoxia. The potential mechanism is partly attributed to inhibiting the gene expressions of HIF-1alpha, mdr1 and MRP1.

OBJECTIVE To explore the advantage of exposure recurrent laryngeal nerve in thyroidectomy. METHODS Exposure the recurrent laryngeal nerve was performed deliberately in 452 cases during thyroidectomy. The larynx was examined with laryngoscope before and after operation. RESULTS Of the 452 cases with 748 exposed recurrent laryngeal nerve, only 14 cases (1.88%) occurred temporary recurrent laryngeal nerve palsy, and 2 cases (0.27%) occurred permanent recurrent laryngeal nerve palsy. CONCLUSION Exposure the recurrent laryngeal nerve during thyroidectomy can protect the nerve from surgery injury.

Purpose To study the clinical features,pathological manifestation and immunohistochemical phenotype and improve the diagnosis and treatment of myoepithelial carcinoma in salivary glands.Methods Histomorphology and immunohistochemical phenotype were analyzed after the sections were stained with routine HE and immunohistochemical methods,and the relevant literatures were reviewed.Results The tumours were predominantly composed of pale-stained clear cells.In some cases,plasma-like cells,epithelioid cells and spindle cells were also seen.The cells were arranged in nest,solid or cords.Mitosis was easily seen,cytological atypia was obvious and necrosis existed in 4 cases.The results of immunohistochemical staining showed that CK was expressed in all cases.EMA was expressed in 8 cases.p63 and CK5/6 were expressed in 11 cases.S-100 was expressed in 10 cases.vimentin was expressed in 4 cases.Calponin was expressed in 2 cases.SMA was expressed in one case.The proliferation index of Ki-67 was 5％ to 40％.Conclusion The histological changes of myoepithelial carcinoma cells are diverse,and pathological and immunohistochemical methods are helpful for improving the rate of right diagnosis.Sugery is the main treatment for myoepithelial carcinoma.

OBJECTIVE: To discuss the safe approach to exposure of the recurrent laryngeal nerve (RLN) in thyroid second operation. METHOD: The data of 153 patients with thyroid second operation were analyzed retrospectively to compare the effectiveness of superior and inferior approach in the exposure of RLN. RESULT: A total of 177 RLNs were exposed in 153 cases. Among those 39 RLNs were exposed by superior approach, 34 by inferior approach after failure of superior approach, and 104 by inferior approach. CONCLUSION In thyroid second operation, inferior approach is a safe and efficient method to expose RLN. Trachea and esophagus are the most important anatomical landmarks to look for the left RLN. Innominate artery and common carotid artery are the most important anatomical landmarks to look for right RLN.
Carotid Artery, Common ; Esophagus ; Humans ; Recurrent Laryngeal Nerve ; surgery ; Reoperation ; Retrospective Studies ; Thyroid Gland ; surgery ; Trachea

OBJECTIVE: To observe the short-term and long-term curative effect of the xenogenic acellular dermal matrix membrane (or joint muscle flap transfer) application used in the 82 cases postoperative tissue shortage repair that after the head neck carcinoma resection. METHOD: To held the 82 cases head neck carcinoma postoperative mucosa shortage repaired after resection by the xenogenic acellular dermal matrix membrane (or joint muscle flap transfer), 65 cases mucosa shortage wound be directly covered by the repair membrane and the other 17 cases mucosa shortage wound be repaired by the tranfered muscle tissue flap with the repair membrane covered; 53 cases underwent additional postoperative radiotherapy between 2-4 weeks and follow-up in 1, 3, 6, 12, 18, 24, 30, 36, 48, 60 months and observed the operation site repair process through the electronic laryngoscope, observed the patients respiration, swallow, phonation function. RESULT: Seventy-seven cases patients operation incision reached I phase healing standard, another 5 cases patients operation incision reached II phase healing standard because of the wound infection and fully-recovered through the local wound drainage,dressing process. All the patients tracheal cannula,the stomach tube be extubated successfully and without the local cicatricial constriction occurred. Seventy-eight cases follow up period reached 1 year including 53 cases who underwent postoperative radiotherapy, 49 cases follow up period reached 3 years including 32 cases who underwent postoperative radiotherapy, 14 cases follow up period reached 5 years including 12 cases who underwent postoperative radiotherapy. The patients with static local lesions discovered no reaction such as exclusion, allergy. CONCLUSION The application of xenogenic acellular dermal matrix membrane (or joint muscle flap transfer used in in the postoperative tissue shortage repair that after the head neck carcinoma resection have several advantage such as comparatively easily implementation, operation safety edge enough,well preserved organ function, comparatively low incidence about the laryngeal stenosis, the short-term and long-term repair effect are all exact.
Acellular Dermis ; Carcinoma, Squamous Cell ; radiotherapy ; surgery ; Female ; Head and Neck Neoplasms ; radiotherapy ; surgery ; Humans ; Male ; Mucous Membrane ; surgery ; Postoperative Period ; Squamous Cell Carcinoma of Head and Neck ; Surgical Flaps ; transplantation ; Wound Healing

OBJECTIVE: To introduce a new transcranial endoscopic approach of glossopharyngeal neurotomy to treat glossopharyngeal neuralgia. METHOD: The clinical data, the operative technique, the operative effect and the result of follow-up of two patients were presented. Through reviewing pertinent literature, we discussed the therapeutic advancement of glossopharyngeal neuralgia and the preponderance of transcranial endoscopic approach of glossopharyngeal neurotomy. RESULT: The two patients both have satisfied outcomes and remained free of disease after surgical treatment. The endoscopic surgery has many significant advantages, such as rigorous operation, little injury, precise therapeutic effect, few complication, and so on. CONCLUSION Transcranial endoscopic approach of glossopharyngeal neurotomy is an ideal method to treat glossopharyngeal neuralgia.
Endoscopy ; Female ; Glossopharyngeal Nerve ; surgery ; Glossopharyngeal Nerve Diseases ; surgery ; Humans ; Male ; Middle Aged ; Neurosurgical Procedures ; methods ; Treatment Outcome

OBJECTIVE: To construct a lentivirus vector of RNA interference (RNAi) of EGFL7 gene and observe its inhibitive role on the invasion of laryngeal cancer cell. METHOD: The effective sequence of siRNA targeting EGFL7 gene was confirmed. Both sense and antisence Oligo DNA of the targeting sequence was designed, synthesized and cloned into the pLV vector,which contained H1 promotor and green fluorescent protein (GFP). The resulting lentivirus vector containing EGFL7 shRNA was called LV-sh EGFL7,and it is confirmed by PCR and sequencing. After that, EGFL7 shRNA was transfected into Hep-2 cells and Western blot was used to test the expression of EGFL7. At last, boyden chamber was used to observe the invasion of the Hep-2 cells. Colony formation assay using a EGFL7 gene silencing on the Hep-2 cell colony forming ability. RESULT: PCR and DNA sequencing demonstrated that the lentivirus RNAi vector of EGFL7 (LV-sh EGFL7) producing EGFL7 shRNA was constructed successfully. The titer of concentrated virus was 5 x 10(8) TU/L. Western blot showed that the expression of EGFL7 was negative in the EGFL7 siRNA Hep-2 cells. And boyden chamber showed the invasive capability of Hep-2 cells transfected EGFL7 siRNA were obviously decreased. EGFL7 gene silencing of cell colony formation rate of cloned Hep-2 cells and compared with empty vector cells, cell cloning and colony formation was significantly reduced. CONCLUSION The lentivirus RNAi vector of EGFL7 was constructed successfully. And EGFL7 silence can inhibit invasion of laryngeal cancer in vitro. After silence EGFL7, Hep-2 cell colony formation was significantly lower, that is, gene expression can be down EGFL7 some extent laryngeal cancer cells inhibited anchorage independent growth capacity.
Cell Line, Tumor ; Endothelial Growth Factors ; genetics ; Gene Expression ; Genetic Vectors ; Humans ; Laryngeal Neoplasms ; genetics ; pathology ; Lentivirus ; genetics ; RNA Interference ; RNA, Small Interfering ; Transfection

OBJECTIVE: To construct the Bmi-1 RNAi expression vector and investigate its influence on the proliferation and invasiveness of laryngeal carcinoma Hep-2 cells. METHOD: The recombinant vshRNA-Bmi-1 plasmid of Bmi-1 RNAi was constructed by the lentiviral expression system, pHelper1.0/pHelper2.0/pGCL2GFP. Bmi-1mRNA and protein expressions of stably transfected laryngeal carcinoma cells were identified tespectively by real-time PCR and Western blot analyses. The changes of the proliferation and invasiveness of laryngeal carcinoma were detected by clone formation test and an invasion assay. RESULT: The Bmi-1 mRNA expressions of stably transfected laryngeal carcinoma Hep-2 cells were significantly decreased. The expression of Bmi-1 protein in laryngeal carcinoma Hep-2 cells was significantly decreased. And the inhibitory rates were 79% and 88% respectively. Whereas the proliferation and invasiveness of Hep-2 cells were significantly reduced. CONCLUSION The Bmi-1 RNAi expression vector were constructed successful. Reduced Bmi-1 expression of Hep-2 cells demonstrated the role of Bmi-1 RNAi in restraining proliferation and invasiveness of laryngeal carcinoma cells.
Cell Line, Tumor ; Cell Proliferation ; genetics ; Genetic Vectors ; Humans ; Laryngeal Neoplasms ; genetics ; metabolism ; pathology ; Neoplasm Invasiveness ; genetics ; Polycomb Repressive Complex 1 ; metabolism ; RNA, Small Interfering ; metabolism ; Transfection

OBJECTIVE: To study the complications in total thyroidectomies and the safety of total thyroidectomy. METHOD: Retrospective analyses 351 cases underwent total thyroidectomy in our department. Preoperative and postoperative electronic laryngoscopy were used to assess the vocal cord function. All patients have serum calcium analysis on the operative day and 3rd and 7th day after operation. RESULT: Temporary hypocalcaemia occurred in 47.01% after total thyroidectomy, but only 15.67% with severe symptom temporarily,and 21.37% with temporary mild numbness. Permanent hypoparathyroidism occurred in 0.85% patients. None with bilateral recurrent laryngeal nerve palsy, and temporary unilateral vocal cord palsy occurred in 1.42% patients and permanent unilateral recurrent laryngeal nerve palsy occurred in 0.56% patients. CONCLUSION Total thyroidectomy is safe with low complications.
Adolescent ; Adult ; Aged ; Child ; Female ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Safety ; Thyroid Neoplasms ; surgery ; Thyroidectomy ; adverse effects ; methods ; Young Adult