Objective To expore the mechanism of low-dose interfone-γ(IFN-γ) influences on differentiation of oligodendrocyte precursor cell. Methods The cerebral cortex samples were obtained from one day old SD rats to form mixed single cell suspensions. After culturing in full medium for 7 to 10 days, succession and differential velocity adherent technique were performed to acquire oligodendrocyte precursor cell and cultured in serum-free medium. IFN-γ, AG490 and Fludarabine were added during the culture of oligodendrocyte precursor cell and reverse transcription-polymerase chain reaction, Western blot and flow cytometry were performed to evaluate the expression of intracellular P27kip1 and its influence on the differentiation of oligodendrocyte precursor cell. Results (1)The expression of P27kip1 mRNA and protein was lower in IFN-γ group than in control group (t=85. 535, P<0. 05;t= 12. 481, P<0. 05), while the expression of P27kip1 mRNA and protein in IFN-γ+AG490 group and IFN-γ+Fludarabine group were both higher than those in IFN-γ group (P<0. 05). (2) The phosphorylation levels of JAK2/STAT1 in INF-γ group were higher than that in the other three groups (P<0. 05). (3) The percentage of myelin basic protein positive cells was (68. 42 ± 2. 53)% in IFN-γ group, lower than that in control group [(88.21 ± 1.97)%](t=10.682, P < 0.05). Myelin basic protein positive cells in IFN-γ + AG490 group were (57. 63 ±2. 75) %, lower than those in the IFN-γ group. The same figure in IFN-γ+Fludarabine group were (79. 53±4. 15)% , higher than those in IFN-γ group (t = 3.957, P<0.05). Conclusions Low-dose IFN-γ can regulate the expression of intracellular P27kip1 through JAK2/STAT1 signal transduction pathway and Fludarabine may participate in this process and improve the differentiation and maturation of oligodendrocyte precursor cell.

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.

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.

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.
Cell Line, Tumor ; Exosomes ; ultrastructure ; Humans ; Laryngeal Neoplasms ; pathology ; Microscopy, Electron, Transmission

[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.

ObjectiveTo study the assistant therapic effect of the Botulinum toxin A (BTA) on tiptoe after cerebral palsy(CP).MethodsMulti point injection of BAT into the musculus gastrocnemius combined with the rehabilitate therapy was used on 35 children(39 feet)with tiptoe after CP.The therapeutic effect was evaluated after a month.ResultsThe efficiency rate is 25.6%,the slight efficiency rate is 71.8%,the ineffective rate is 2.6%.There is no toxic or side effect was observed.Conclusions The BTA can relieve spasm of musculature and redress abnormality from convulsing of musculature.

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.
Animals ; Apoptosis ; Cisplatin ; pharmacology ; Cochlea ; metabolism ; Guinea Pigs ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Spiral Ganglion ; drug effects ; metabolism ; bcl-2-Associated X Protein ; metabolism

OBJECTIVE To investigate the factors influencing the effects of disinfection of HBV on dental hand piece. METHODS After polluted with HBV positive serum,the dental hand piece was disinfected with sterilizer at the condition of 134℃ for 5 minutes according different treatment procedures.The samples were then taken and detected HBsAg by the automatic enzyme immune analyzer using the third generation EIA agent.The efficacy of disinfection of HBV was presented by the reaction of HBsAg detection. RESULTS If the dental hand piece was sterilized directly not cleansed using clean water after pollution and dried in room temperature,there were 96.87% of the samples being HBsAg positive,and the polluted but dried hand piece,even cleansed there were 87.5% of positive result, even if prolonged the time of sterilization and the frequencies of vacuumization,the HBsAg positive rate of the sample was still up to 56.25%.But if the hand piece was irrigated and cleansed immediately after pollution and sterilized,all samples were HBsAg negative. CONCLUSIONS Whether or not cleansed immediately is the most important factor influencing the efficacy of hand piece sterilization after pollution.

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.
Bone Conduction ; Cartilage ; Cohort Studies ; Ear Auricle ; Endoscopy ; Hearing ; Hearing Tests ; Humans ; Myringoplasty ; Postoperative Period ; Retrospective Studies ; Treatment Outcome ; Tympanic Membrane ; Tympanic Membrane Perforation ; surgery ; Tympanoplasty

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.