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 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.

[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.

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 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

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.

This study is to examine the effects of NNIspm-mediated cellular senescence of HepG2 cells and elucidate its potential molecular mechanism. Cellular senescence was detected with senescence-associated beta-galactosidase staining. Cell cycle distribution, intracellular fluorescence intensity and accumulation of intracellular reactive oxygen species (ROS) were detected by high content screening (HCS). Protein expression was detected by Western blotting. Polyamines content was analyzed by high performance liquid chromatography (HPLC). The results demonstrated that NNIspm significantly induced HepG2 cells senescence. This effect was due to the decrease of intracellular polyamines, the arrest at G0/G1 phase and an increase of ROS level. The molecular senescence marker p21 increased significantly after NNIspm treatment. In contrast, the protein expressions of Cyclin E and CDK2 were obvious down-regulation. The results indicated that cellular senescence induced by NNIspm was one of its antitumor mechanisms.

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.

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.

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.