Effectiveness of cisplatin on the expressions of Bcl-2 and Bax in cochlea and spiral ganglion cells of guinea pigs / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To investigate the effectiveness of cisplatin on the expressions of Bcl-2 and Bax in cochlea and spiral ganglion cells (SGC) of guinea pigs.@*METHOD@#Twenty guinea pigs were randomly divided into cisplatin (n = 10) and control groups (n = 10). Cisplatin group were administrated with a dose of intraperitoneal injection of 16 mg/kg, while the control group were received intraperitoneal injection of normal saline as placebo. Before and 7 days following injections, the ototoxic effect was measured with distortion product otoacoustic emission (DPOAE). Bcl-2, Bax in cochlea were detected by Western Blot. Immunohistochemical staining was used to detect the protein levels of Bcl-2 and Bax in spiral ganglion cells.@*RESULT@#In control and cisplatin group, Bcl-2 protein levels were 0.727 8 ± 0.016 9 and 0.467 6 ± 0.020 1, Bax protein levels were 0.384 8 ± 0. 0217 and 0.735 6 ± 0.022 3 in cochlea respectively, both P < 0.01. In Control and cisplatin group, the grey values of Bcl-2 in SGC were 99.00 ± 2.42 and 149.80 ± 2.37 respectively, the grey values of Bax were 154.50 ± 2.80 and 104.50 ± 3.09 respectively, both P < 0.05.@*CONCLUSION@#Decreased expression of Bcl-2 and increased expression of Bax may be involved in cisplatin-induced apoptosis in cochlea and SGC of guinea pigs.

The Protective Effects of Intratympanic Dexamethasone to Prevent Cisplatin Ototoxicity / 听力学及言语疾病杂志

Objective To investigate the effectiveness of intratympanic dexamethasone injection as a protec-tion agent against cisplatin ototoxicity.Methods Fifty five healthy guinea pigs were randomly divided into four groups.The guinea pigs groups received (group I)0.9% NaCl solution(16 ml/kg)intraperitoneally (IP),(group II)dexamethasone (10 mg/ml)intratympanically(IT),(group III)cisplatin (16 mg/kg)only intraperitoneally(IP) and (group IV)dexamethasone (10 mg/ml)IT followed after 1h by 16 mg/kg cisplatin.Before and 7 days following injections,the ototoxic effect was measured with auditory brainstem response(ABR).The contents of malondiade-hyde(MDA)and activities of superoxide dismutase(SOD)were also measured.Results ABR thresholds before and after dosing were 28.50±4.74,29.00±3.94 dB nHL and 28.67±5.82,31.33±5.81 dB nHL in group I and group II,respectively,with no significant differences (P>0.05).The contents of MDA and activities of SOD were 2.01 ±0.07 mmol/L ,234.10±13.09 U/ml and 2.06±0.09 mmol/L,233.20±13.24 U/ml in the group I and group II, respectively,with no significant differences (P>0.05).ABR thresholds after dosing were 55.33±4.81,and 40.67 ±3.72 dB nHL in the group III and group IV,which were significantly increased,but ABR thresholds in the group IV were significantly lower than that in the group III (P<0.05).The contents of MDA were 5.74±0.17,3.51± 0.18 mmol/L in the group III and group IV,which were significantly increased.SOD activities were 107.90 ± 14.21,and 162.70±11.25 U/ml,which significantly decreased.Nevertheless,in the group IV dexamethasone rela-tively reduced the contents of MDA(P<0.001)while increased the activities of SOD(P<0.001)compared with group III.Conclusion Intratympanic dexamethasone injection presents as an effective treatment modality to prevent cisplatin ototoxicity without any ototoxic effect.

Comparison of methods for isolating exosomes derived from laryngocarcinoma Hep-2 cells / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To provide a reference about choosing the methods of isolating exosomes derived from tumor cells including laryngocarcinima Hep-2 cells by comparing advantages and defects of two methods of isolation and extraction exosomes.@*METHOD@#Previously, laryngocarcinoma Hep-2 cells were cultivated massively, then the cells were processed with hot shock in 42 degrees C for 1 h. Sucrose density gradient centrifugation ultrafiltration (method 1): cells culture supernatant 90 ml was gathered, the supernatant was clarified through a 3/0.8 μm small filter to remove impurities and fragments which in larger diameter. Then the filtering fluid was concentrated and purified through sucrose density gradient centrifugation and ultrafiltration, the concentrated fluid was obtained. Exosome Isolation Kit (method 2): cells culture supernatant 4 ml was gathered, the solutions of the kit were added into the supernatant in proper sequence, then filtered by the special column, the concentrated fluid was obtained. Both products are observed by high resolution transmission electron microscopy.@*RESULT@#Both methods could isolate and extract exosomes feasibly. In single high power view of transmission electron microscopy, exosomes of method 1 disperse better, but lower density, and more impurity in background, exosomes of method 2 arrange closer, higher density, and less impurity.@*CONCLUSION@#Exosome isolation Kit require less supernatant, cost less time, process procedure briefly, harvest higher yield. It may become a new option of isolating exosomes derived from Laryngocarcinoma Hep-2 cells.

Expressions of tumor angiogenesis related factors CD105 and Galectin-3 in laryngeal squamous cell carcinoma and biological assessment / 中国耳鼻咽喉头颈外科

[ABSTRACT]OBJECTIVETo investigate the expressions of Endoglin (CD105) and Galectin-3 protein in laryngeal squamous cell carcinoma (LSCC) as well as the relationship between their expressions and the clinicopathological factors of LSCC.METHODSThe expressions of CD105 and Galectin-3 protein were detected in 76 samples of LSCC and 25 normal laryngeal tissues (NLT) by immunohistochemical staining (S-P).RESULTS 1.The mean of Microvessel density (MVD) value marked by CD105 in LSCC was 10.33±2.29, which was significantly higher than that in NLT (1.20±1.04) (P<0.05). The expression of MVD marked by CD105 (CD105-MVD) was correlated with histological grading, T stage, clinical stage, lymph node metastasis, recurrence and prognosis in LSCC (P<0.05). 2.The positive expression rate of Galectin-3 protein in LSCC was 86.84%, which was significantly higher than that in NLT (36%)(P<0.05). The expression of Galectin-3 was correlated with T stage,clinical stage, lymph node metastasis, recurrence and prognosis in LSCC (P<0.05). 3.There was a positive correlation between CD105 and Galectin-3 protein. 4.Survival analysis indicated that the expressions of CD105 and Galectin-3, histological grading, lymph node metastasis,T stage and recurrence were independent factors for tumor prognosis in LSCC (P<0.05). CONCLUSIONThe expressions of CD105 and Galectin-3 protein have a positive correlation in LSCC. They may play important roles in the tumorigenesis, malignant progression and poor prognosis of LSCC. Combined detection of them may be great value in diagnosis and predicting prognosis of LSCC.

The effectiveness of endoscopic tragus cartilage-perichondrium myringoplasty in the treatment of large tympanic membrane perforations / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To evaluate effectiveness of endoscopic tragus cartilage-perichondrium in the treatment of large tympanic membrane perforations.@*METHOD@#Patients with large tympanic membrane perfortations who had underwent cartilage-perichondrium myringoplasty were retrospectively retrieved from our department. Those with a follow-up of equal to or greater than 12 months after surgery were included in the study. Hearing test results were reported using a four-frequency air conduction and bone-air conduction gap. Patients were labeled as treatment success if the tympanic membrane was intact without lateralization or anterior blunting after surgery.@*RESULT@#Of the 35 patients with large tympanic membrane perforations treated by endoscopic tragus cartilage-perichondrium, 33 (94.3%) were treatment success. There was no graft lateralization, anterior blunting, neocholesteatoma, and sensorineural hearing loss in these patients after surgery. The air conduction and bone-air conduction gap before surgery in the study cohort were (43.8 ± 5.7) dB and (28.5 ± 3.1) dB, respectively. Postoperative air conduction and bone-air conduction gap of the cohort were (31.4 ± 6.4) dB HL and (16.2 ± 4.1) dB, respectively. The postoperative air conduction and bone-air conduction gap decreased significantly after surgery (P < 0.01). Overall postoperative air-bone gap in 27 of the 35 patients (77%) were less than 20 dB after surgery.@*CONCLUSION@#Endoscopic cartilage-perichondrium myringoplasty is an effective procedure in the treatment of large tympanic membrane perforations.

The influence of sublingual immunotherapy on Th2 type immunoreaction in patients with allergic rhinitis / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To explore the influence of sublingual immunotherapy on Th2 type immunoreaction in patients with allergic rhinitis within 12 months' treatment.@*METHOD@#Forty three patients with allergic rhinitis were followed-up within 12 months after receiving sublingual immunotherapy. The changes of clinic symptoms and immunoreaction were analyzed. VAS score was applied to evaluate the efficacy of clinic treatment. TIgE and sIgE in serum were tested to evaluate the systemic immunoreaction. Eosinophil counting in nasal discharge was applied to evaluate the local nasal mucosa immunoreaction.@*RESULT@#VAS scores were decreased significantly one month after the starting point of treatment, companied by the decrease of EOS count in nasal discharge (P < 0.05). The changes of TIgE and sIgE in serum within 12 months of sublingual immunotherapy were not significant (P < 0.05). The levels of IL-4 and IL-13 in nasal lavage were decreased significantly after 12 months' treatment.@*CONCLUSION@#The bias of Th2 type immunoreaction existed in nasal mucosa immune system in patients with allergic rhinitis could been rectified partially after receiving sublingual immunotherapy for 12 months, but the significant changes were not observed in systemic immune system.

Melanoma antigen A3 sourced from laryngeal cancer was expressed in mouse melanoma cell model / 中国免疫学杂志

Objective:To construct tumor cell model by determination of the pIRES2-EGFP/MAGE-A3 eukaryotic expression plasmid expressing steadily in mouse melanoma B16 cells.Methods:The pIRES2-EGFP/MAGE-A3 eukaryotic expression plasmid being constructed from the melanoma-associated antigen A3 genes sourcing laryngocarcinoma in advance was translated into the mouse melanoma B16 cells under the mediation of lipofectamine,and the positive clones were detected with G418.The expression of enhanced green fluorescent protein( EGFP) and MAGE-A3 mRNA in positive clones were detected by fluorescence microscopy and fluorescence quantitative PCR ( qRT-PCR ) assay, respectively.Results:The pIRES2-EGFP/MAGE-A3 eukaryotic expression plasmid has been transfected into B16 cells successfully, the green fluorescence of fusion protein expression was found, and MAGE-A3 mRNA transcription in B16 cells expressions were detected in positive clones.Conclusion:The pIRES2-EGFP/MAGE-A3 eukaryotic expression plasmid has been transfected effectively and expressed stably by liposome method in the B16 cells.The expression of MAGE-A3 tumor cell model has been successfully established,which provide data for the study of laryngocarcinoma immunotherapy.

Efficacy of intratympanic dexamethasone combined with prednisone on idiopathic sudden sensorineural hearing loss / 实用医学杂志

Objective To evaluate the therapeutic efficacy of intratympanic dexamethasone injections combined with prednisone in patients with idiopathic sudden sensorineural hearing loss. Methods A total of 71 patients diagnosed with sudden hearing loss were treated with intratympanic dexamethasone injections plus prednisone (B group) or prednisone alone (A group). Hearing was evaluated by pure tone audiogram performed before initial treatment and at 4 weeks following the final treatment. Results The total recovery rate after the treatment was 81.8% in the B group and 55.3% in the A group. The diflference between two groups was statistically significant (P < 0.05). Conclusion The present study suggests that sudden sensorineural hearing loss patients treated with intratympanic dexamethasone combined with prednisone have a higher likelihood of hearing recovery than those treated with prednisone alone.

Construction of the superantigen SEA transfected laryngocarcinoma cells / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To construct an eukaryotic expression vectors containing superantigen staphylococcal enterotoxin A (SEA) gene, and to identify its expression in laryngeal squamous carcinoma cells.@*METHOD@#SEA full-length gene fragment was obtained from ATCC13565 genome of the staphylococcus, referencing standard strains producing SEA. Coding sequence of SEA was artificially synthetized. Than, SEA gene fragments was subcloned into eukaryotic expression vector pIRES-EGFP. The recombinant plasmid pSEA-IRES-EGFP was constructed and was transfected to laryngocarcinoma Hep-2 cells. Resistant clones were screened by G418. The expression of SEA in laryngocarcinoma cells was identified with ELISA and RT-PCR method.@*RESULT@#The subclone of artificially synthetized SEA gene was subclone to eukaryotic expression vector pires-EGFP. Flanking sequence confirmed that SEA sequence was fully identical to the coding sequence of standard staphylococcus strains ATCC13565 in Genbank. After recombinant plasmid transfected to laryngocarcinoma cells, the resistant clones was obtained after screening for two weeks. The clones were selected. The specific gene fragment was obtained by RT-PCR amplification. ELISA assay confirmed that the content of SEA protein in supernatant fluid of cell culture had reached about Pg level.@*CONCLUSION@#The recombinant eukaryotic expression vector containing superantigen SEA gene is successfully constructed, and is capable of effective expression and continued secretion of SEA protein in laryngochrcinoma Hep-2 cells after recombinant plasmid transfected to laryngocarcinoma cells.

Study of simultaneous audition reconstruction to chronic suppurative otitis media in the period of infection / 中国医师进修杂志

Objective To investigate the possibility and surgical effect of simultaneous tympanoplasty to chronic suppurative otitis media in the period of infection. Methods Forty-eight cases (48 ears) with chronic suppurative otitis media in the period of infection (31 with cholesteatoma, 17 with caries) were underwent simultaneous Wullstein Ⅱ and Ⅲ tympanoplasty on the complete elimination of the lesions (typical or modified mastoidectomy). Results All eases had dry ears within 4-10 weeks with average of 7 weeks. The air-bone gap within 10 dB was in 11 eases, 15 to 20 dB in 25 cases, 25 to 30 dB in 9 eases, no change or worse in 3 eases. Conclusions Infection is not the absolute eontraindication to the tympanoplasty in treating chronic suppurative otitis media, Wullstein Ⅲ tympanoplasty plus mastoid cavity obliteration and eonchaplasty is a suitable choice to treating chronic suppurative otitis media on the complete elimination of lesions and reconstruction of the ventilation system among mastoid cavity, tympanum and eustachian. The malfunction of eustachian is the main eanse to failure of surgery.

Establish and evaluate allergic rhiultis model in guinea pigs with Toluene-2,4-diisocyanate / 中国医师杂志

Objective To understand the changes of allergic rhinitis from immunology,pathophysiology and morphology method were applied.Methods Toluene 2,4-diisocyanate was dissolved with florence oil to final concentration 10%,this solution was dropped into the nasal vestibules of animals to induce sensitization process.8 guinea pigs were prepared as allergic rhinitis with 10% Toluene 2,4-diisocyanate,and other 8 animals were used as blank controh.After model successfully established,the blood were obtained to determine RBC-C3b receptor rosette rate and RBC-imuuunocomplex rosette rate.The nasal mucosas were obtained from 2 groups,distribution and changes of substance P in nasal mucosa were observed with the application of immunohistochemical staining.The content of blood histamine was determined with ELASA method.The pathological changes of nasal mucosas were observed with the application of light microscope and transmission electron microscope.Results The behavior scores of the modeling animals were significantly higher than that of controls(P＜0.01).The value of RBC-C3b reeeptor rosette rate and RBC-imuuunoeomplex rosette rate of the modeling animals were significantly decreased than that of the controls(P＜0.05).Substance P presented in the normal nasal mucosal epithelial cells,epithelium cells of blood vessels,glandular cells and its duets,the staining density and the positive staining cells in the same aero in modeling group significantly increased,compared with controls.The counts of substance P-positive cells of control group were less than those of modeling group(P＜0.05).The content of blood histamine of the modeling animals were significantly increased compared with the controls(P＜0.01).The structure of false multiple coat cilium columnar epithelial cells in nasal mucosa of control group were successive,intact,and distinct.There were normal mucosal epithelium,lamina propria and submucosa.But modeling group showed that mucosal epithalamiums were damaged and shed,goblet cell proliferated,squamous metaphase,and epithelial necrosis happened,serous glands in lamina propria vigorously proliferated,blood vessels expanded,tissue edema formed,plenty of inflammatory cell such as eosinophil and mast cells were more in number and infiltrated.The structure of epithelial eells,cilia and mierotubule of nasal mucosa of control group were regular and distinct,there were abundant cellular organs in cytoplasm.Eosinophil cells were intact.But iu model group,mucosal epithalamium,cilia and its microtubule,mierovillus,goblet cell were damaged,the cell bulk and nucleus changed,mast cells and eosinophil cells changed,blood vessels expanded,serous glands vigorously proliferated.This morphological change was roughly identical to clinical manifestation of allergic rhinitis.Conclusion Toluene-2,4-diisocyanate can be used to establish allergic rhinitis model in guinea pigs,and some ehanges of the allergic rhinitis in model guinea pig were similar with clinic observation.

Measurement of components of the phospholipid of the surfactant in irrigating fluid from the nasopharynx of patients with chronic sinusitis / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To investigate the effect of chronic sinusitis on components of the phospholipid of nasopharyngeal surfactant, and to study biochemical component of phospholipid of surface active substance.@*METHOD@#The concentrations of surfactant in nasopharyngeal irrigating fluid were implemented in normal controls and patients with chronic sinusitis. Components of phospholipid such as Phosphatidylserine, Phosphatidylethanolamine, Phosphatidylcholine and Sphingophospholipid were measured by the high-performance liquid chromatograph.@*RESULT@#Results showed as follows (1) There was surfactant in nasopharynx. 4 compositions of phospholipid could be measured. (2) Compared with controls, Phosphatidylserine signficantly decreased in patients with chronic sinusitis (P < 0.05). (3) Only Phosphatidylserine signficantly decreased between sinusitis III stages and controls (P < 0.05). The rests had no signficant difference between chronic sinusitis' stages and controls, and among stages. But as the chronic sinusitis' stages proceeded, proportion of Phosphatidylserine may decreased.@*CONCLUSION@#(1) There is surfactant in nasopharynx, nasopharyngeal surfactant is made of Phosphatidylserine, Phosphatidylethanolamine, Phosphatidylcholine and Sphingophospholipid. The proportion of Phosphatidylcholine shows most, and determines biochemical effect of nasopharyngeal surfactant. (2) chronic sinusitis may cause decrease of some components of nasopharyngeal surfactant. (3) As the chronic sinusitis' stages proceed, the proportion of some phospholipids progressively decrease. Which, above assessed, may cause the change of surfactant in eustachian tube, and cause dysfunction of middle ear and eustachian tube.

Clinical significance of regional excision of parotid pleomorphic adenoma / 中国耳鼻咽喉头颈外科

OBJECTIVE To investigate the feas-ibility of preserving parotid duct in regional excision of pleomorphic adenoma. METHODS 32 patients with parotid pleomorphic adenoma were treated by regional excision with the preservation of the parotid duct. RESULTS No relapse occurred during 3-6-year follow-up period. Transient facial nerve palsy occurred in only 3.1%(1/32) of patients and the drainage of the parotid duct on the surgical side was preserved. CONCLUSION Remaining function of the parotid gland function was preserved and the facial nerve was well-protected in the regional excision of pleomorphic adenomas, especially for patients with small pleomorphic adenomas.