The Effects of Ouabain on the Inner Ear Glial Cells in Mouse / 听力学及言语疾病杂志

To study the effects of ouabain on the inner ear glial cells, and to lay the foundation for the study of stem cell transplantation in the treatment of sensorineural hearing loss.Methods Sixty adult female SPF grade CBA / J mice were randomly divided into experimental group and control group, with 30 mice in each group.Animals in the experimental group received 3mM ouabain via the round window membrane, while mice in control group received normal saline.The mice were sacrificed at 7 days, 14 days and 30 days after the administration,respectively.Immunofluorescence histochemical staining was used to detect the inner ear glial cells in spiral ganglion.Results Some inner ear glial cells survived in the spiral ganglion of the experimental group, while with decreased numbers and disorganized structure compared to those of in the control group.Comparing to those of in the control group, the number and density of inner ear glial cells in the experimental group were significantly decreased from 7 days afterouabain administration,further decreased at 14 days and reduced to the lowest at 30 days after ouabain administration, the differences between the 2 groups were statistically significant (P < 0.05).Among the experimental group, the number of inner ear glial cells at 30 days was significantly decreased when compared to those of at 7 days and 14 days, respectively.Conclusion Application of ouabain to mouse inner ear via the round window membrane leads to an acute and progressive direct damage to the inner ear glial cells in the spiral ganglion.

Expression and Relation of TERT and AP-1 in Laryngeal Carcinoma / 听力学及言语疾病杂志

Objective To investigate telomerase reverse transcriptase (TERT ) and acticator protein 1(AP -1) expression and it's corroloation in laryngeal carcinoma tissue .Methods 24 human laryngeal carcinoma tissue were analised by RT -PCR and quantum -dot based immunofluorescence assay for the expression of TERT and AP -1 . Results The expression of TERT mRNA and the expression of c -Fos ,c-Jun in laryngeal carcinoma were postive‐ly related .Correlation coefficient was 0 .574 and 0 .809 ,respectivly(P<0 .01) .Conclusion TERT and AP -1 were expressed at high levels and positively correlated in human laryngeal carcinoma .

Effects of Long Term Injection of Sodium Salicylate on the ABR and Expression of GAD67 in Rat Inferior Colliculus / 听力学及言语疾病杂志

Objective To observe the effects of long term injection sodium salicylate on the auditory brain-stem response(ABR)and expression of glutamic acid decarboxylase -67(GAD67) in rat inferior colliculus .Methods Eighteen healthy Wistar rats were randomly divided into three groups :the sodium salicylate group (intramuscular injection of 10% sodium salicylate ,175 mg/kg ,twice daliy for 28 days) ,the saline group (intramuscular injection with saline on same does at the same time) ,the control group (without any treatment) .The rats received ABR after modeling ,then were decapitated and inferior colliculus tissues were stripped .Western blot was used to study the dif-ferent expression of GAD67 protein levels in the three groups .Results Compared with the saline group and control group ,ABR thresholds of the sodium salicylate group were significantly elevated and latency of wave Ⅲ was aslo sig-nificantly prolonged(P0 .05) .The inferior colliculus GAD67 protein expression level of sodium salicylate group was significantly higher than the saline group and control group(P0 .05) .Conclusion Long term injection of sodium salicylate can cause a change in the inferior colliculus of GAD67 protein expression and the up regulation of GAD67 expression may occur as a com-pensatory response to increase inhibiting effect .The change of GAD67 protein expression is likely as a compensatory and regulatory mechanisms for sodium salicylate ototoxicity .

Observation on morphology and nerve growth factor expression in nasal mucosal of depression rats / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To establish the rat model of depression and observe its morphological changes and nerve growth factor (NGF) expression in nasal mucosal.@*METHOD@#Thirty SD rats were divided into depression model group and normal controls with 15 rats in each group. The rat depression model were made by application of chronic unforeseeable medium stress stimulating. Behavior change were observed with open box experiment(open field) and humoral consumption-experiments (sucrose consumption), After being sacrificed, the nasal mucosal were taken for HE and NGF immunohistochemical study. The difference of nasal mucosal NGF protein between two groups were examined with medical digital image analysis technology.@*RESULT@#There were no swelling and inflammatory cells infiltrating in nasal mucosal of normal group, but 53% nasal mucosal of depression model were observed such morphological changes. Th NGF immunohistochemical staining were negative in normal group but positive in 53% depression model group. There were significant differences between two groups (P < 0.01).@*CONCLUSION@#Depression rats may appear rhinitis symptoms under long-term stress stimulating. NGF may exert immunoregulatory effects, stimulate the immune cell proliferation and gathering, induce various media release, increase the sensitivity of nerve fibers itself and the producing of neuropeptide, promote the formation of nerve source inflammation and increase the reactivity of nasal mucosa.

The analysis of AllergyScreen results among allergic patients of different age groups / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To investigate the positive rates of some allergens in different age groups of allergic rhinitis patients, and to analyze the relationship between serum specific immunoglobulin E(sIgE) and total immunoglobulin E(IgE).@*METHOD@#Six hundred and fifty-three patients with allergic rhinitis, tested by AllergyScreen system, were enrolled in our research.@*RESULT@#The positive rate of sIgE in all patients was 85.5%. The positive rate of dust mite was the highest with the result of 76.3%, and the positive rate of feline or dogs hair was the lowest of 20.3%. The statistical significance of different positive rate of some allergens were found in the different age groups. All the patients were divided into two groups, one was younger than 18 years old group, which including 18 years old and another was older than 18 years old group. Except Mx, the positive rate of other allergens existed statistical difference. The level of serum total IgE could not reflect the positive rate of sIgE completely.@*CONCLUSION@#It is an effective and reliable method of the diagnosis of allergic rhinitis to combine the analysis of total serum IgE and sIgE levels with patients clinical manifestations. Different strategies should be employed for the prevention and treatment of allergic rhinitis according to their ages.

Meta-analysis of the effects of radiotherapy and surgery on carcinoma of the middle ear / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To evaluate the effects of radiotherapy and surgery on carcinoma of the middle ear.@*METHOD@#A review of five published literatures was conducted according to defined selection criteria by the Review Manager 5.0 statistical software.@*RESULT@#There were no systematic reviews or large-scale RCTs between radiotherapy and symptomatic treatment containing surgery and radiotherapy for carcinoma of the middle ear.@*CONCLUSION@#Radiotherapy and symptomatic treatment for carcinoma of the middle ear have no obvious differences. The radiotherapy is the first choice for the treatment of squamous cell carcinoma of the middle ear.

Expression and significance of SP-A in nasal mucosa of allergic rhinitis and nasal polyp / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#Surfactant protein A (SP-A) is protein that appears to play an important role in mammalian first-line host defense. The objective of this study was to immunolocalize SP-A in human sinonasal tissue.@*METHOD@#Eleven cases of allergic rhinitis, fifteen cases of polyp and seven cases of normal middle turbinate were studied with immunohistochemistry and immunofluorescence method to detect the expression of SP-A.@*RESULT@#The expression of SP-A in allergic rhinitis and polyp were dramatically higher than that in controls (P 0.05). The result demonstrated that SP-A was positively correlated with eosinophils within the basement membrane of epithelium (R = 0.81, 0.55). In the result of immunofluorescence, there was significantly higher expression SP-A in nasal mucosa of allergic rhinitis and nasal polyp than that in control group (P < 0.05).@*CONCLUSION@#SP-A is likely to play key roles in the inflammatory reaction process of allergic rhinitis and polyp. Its secretion in the upper airway indicates that future studies may allow manipulation of this protein and development of novel treatments for sinonasal pathology.

Expression and significance of SP-A in nasal mucosa of allergic rhinitis and nasal polyp / 临床耳鼻咽喉头颈外科杂志

Objective: Surfactant protein A (SP-A) is protein that appears to play an important role in mammalian first-line host defense. The objective of this study was to immunolocalize SP-A in human sinonasal tissue. Method:Eleven cases of allergic rhinitis, fifteen cases of polyp and seven cases of normal middle turbinate were studied with immunohistochemistry and immunofluorescence method to detect the expression of SP-A. Result:The expression of SP-A in allergic rhinitis and polyp were dramatically higher than that in controls(P0.05). The result demonstrated that SP-A was positivly correlated with eosinophils within the basement membrane of epitheli-um(R=0.81,0.55). In the result of immunofluorescence, there was significantly higher expression SP-A in nasal mucosa of allergic rhinitis and nasal polyp than that in control group(P<0.05). Conclusion:SP-A is likely to play key roles in the inflammatory reaction process of allergic rhinitis and polyp. Its secretion in the upper airway indicates that future studies may allow manipulation of this protein and development of novel treatments for sinonasal pathology.

Analysis about the results of allergy screen in 134 cases of allergic rhinitis / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To analyze the results of serum in allergic rhinitis and investigate the specific clinic allergen and serum immunoglobulin E (IgE) levels.@*METHOD@#Allergy Screen method was used to detect the specific allergen and total serum IgE level of 134 cases of Allergic rhinitis.@*RESULT@#The dust mite was the most common allergen in inhalation group in 134 cases of allergic rhinitis, the positive rates was 90%; then were donly, feline and scurfy fungus, the positive rates were 16%, 9%. The positive rates of total IgE was 54%. The serum IgE levels between 100 to 200 kU/L, there was 21 cases together, but there existed 7 negative cases. There were 51 cases' IgE levels more than 200 kU/L, the rates was 70.8%, but there still existed 4 negative cases.@*CONCLUSION@#Allergy screen method can find relevant allergen and provide basis for the prevention and treatment of allergic disease.

Research on DPOAE of guinea pigs treated with gentamicin / 生物医学工程学杂志

This study was designed to disclose the specific changes of distortion product otoacoustic emission (DPOAE), including DP-gram and I/O-gram in guinea pigs treated with GM for 10 and 17 days. Forty guinea pigs were divided into four groups:group I (treated with GM for 10 days), group II (treated with normal saline for 10 days), group III (treated with GM for 17 days), group IV (treated with normal saline for 17 days). DPOAE including DP-gram and DP-I/O was recorded by use of CELESTA 503 Cochlear Emission Analysis and the out hair cell loss was numerated. (1) The amplitude in group I showed significant reduction at frequencies of 4, 6, 8 kHz. There were significant differences, compared to group II (t=2.52, 1.92, 2.10, P<0.05). The amplitude in group III also decreased at frequencies of 3, 4, 6, 8 kHz. There were also significant differences compared to group IV (t=3.27, 2.81, 2.92, 3.13, P<0.01). The amplitude of DP-I/O in group I and group III at different frequencies and stimulus levels showed reduction. (2) At the level L1 <60 dB SPL, the stimulus levels to elicit a 0 dB response in group I and group III were higher than those in corresponding control groups, respectively. (3) The mean of threshold shift was 5.0+/-3.8 dB in group I and 25.0+/-6 dB in group III at 8 kHz frequency. (4) The out hair cell loss was 22.16% in group I and 48.36% in group III, both showed significant difference as compared to controls, respectively (P<0.01). The employment of DPOAE can be a useful tool in monitoring ototoxicity of GM for its sensitivity and specificity in guinea pigs treated with GM. The changes in DPOAE were consistent with those in histology.

The effect of shRNA targeting hTERT on telomerase and the expression of PCNA and Caspase-3 in nasopharyngeal carcinoma cells / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#Analysis of the correlation between telomerase and expression of its related proteins may provide insight into the molecular mechanism of nasopharyngeal carcinogenesis. Here, we investigate the effect of short hairpin RNA (shRNA) specific for human telomerase reverse transcriptase (hTERT) mRNA on the expression of proliferating cell nuclear antigen (PCNA) and Caspase-3 in nasopharyngeal carcinoma (CNE) cells.@*METHOD@#shRNA expression vectors targeting the mRNA of hTERT were constructed. Cells were treated with the constructed expression vectors and telomerase activity was measured by telomeric repeat amplification ELISA (TRAP-ELISA). Cell viability was examined using the MTT assay. Cell apoptosis was detected by TUNEL method. The expression of PCNA and Caspase-3 proteins, was determined by Western blotting.@*RESULT@#shRNA specific for hTERT mRNA significantly inhibited telomerase activity, suppressed cell viability and induced apoptosis in CNE cells. In addition, the expression of PCNA was inhibited, while the expression of Caspase-3 was up-regulated.@*CONCLUSION@#Our results suggest that shRNA directed against hTERT inhibits cell viability by regulating telomerase activity and its related protein expression in NPC cells. Therefore, RNA-interfering technology may be a promising strategy for the treatment of nasopharyngeal cancer.

Effect of short hairpin RNA on growth and proliferation of human bone marrow mesenchymal stem cells / 中国组织工程研究

BACKGROUND:Tetramethylpyrazine (TMP) can inhibit the expression of vascular endothelial growth factor (VEGF), but it is uncertain that TMP inhibit the growth and proliferation of HL-60 leukemic cells induced by VEGF.OBJECTIVE:To observe the effect of TMP on the proliferation of HL-60 leukemic cells induced by VEGF.DESIGN:Repetitive measurement and observation.SETTING:School of Medicine, Wuhan University of Science and Technology.MATERIALS:The experiment was carried out in the Molecular Biology Laboratory Center, School of Medicine, Wuhan University of Science and Technology from March to June in 2007. Human leukemic cell line HL-60 cells were purchased from Shanghai Institute of Cell Biology. TMP hydrochloride injection was produced by Wuxi Seventh Pharmaceutical Products Limited (Lot number:011014), protamine sulfate injection was produced by Shanghai First Biochemical Pharmaceuticals (Batch number:010302), and immunohistochemistry kit was purchased from Boster company.METHODS:①Human leukemic cell line HL-60 cells at log phase were used for the experiments. Cells were treated with 100 μg/L VEGF, and then TMP at final concentrations of 1.5, 15, 150 mg/L was added into culture medium. While the cells in medium without TMP were taken as blank control group, and the cells in medium with 20 mg/L protamine as positive control group. Meanwhile cells without treatment of VEGF were served as VEGF control group. After cells were incubated for 48 hours, the growth inhibiting rate of HL-60 cells was detected by MTT assay.②After HL-60 cells were treated with TMP at the final concentrations of 1.5, 15, 150 mg/L for 24 hours, the protein expression of VEGF in HL-60 cells was examined by SP immunohistochemistry.MAIN OUTCOME MEASURES:①Growth inhibiting rate of HL-60 cells.②Protein expression of VEGF.RESULTS:①Growth inhibiting rate of HL-60 cells:After HL-60 cells induced by VEGF were treated with 15 and 150 mg/L TMP, the absorbance value was significantly lower than that in VEGF control group (P < 0.05).②Protein expression of VEGF:After HL-60 cells were treated with TMP for 24 hours, the protein expression of VEGF was down-regulated with increasing TMP concentration in a dependent manner. Significant differences were observed in the protein expression of VEGF between cells treated by TMP and the controls (P < 0.01).CONCLUSION:shRNA, by targeting hTERT mRNA, has no noticeable influence on growth and proliferation of hMSCs, and might be safe for the somatic cells which are normal but do not express hTERT.

Expression and significance of Eotaxin and Eotaxin-2 in nasal polyposis and nasal polyp tissue / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To study the expression of Eotaxin and Eotaxin-2 in nasal polyposis and nasal polyp and explore the different mechanism between polyposis and polyp.@*METHOD@#Fifteen cases of nasal polyposis, thirteen cases of polyp and eight cases of normal middle turbinate were studied with immunohistochemical SP method to detect the expression of Eotaxin and Eotaxin-2.@*RESULT@#There were significant differences between either two groups about the expression of Eotaxin-2 (P 0.05).@*CONCLUSION@#Both of Eotaxin and Eotaxin-2 likely play key roles in the inflammatory reaction process of nasal polyposis and polyp. The different expression of Eotaxin-2 between nasal polyposis and polyp demonstrates it may be one of the main causes in the different mechanism of the two diseases.

Expression of PML in carcinogenesis of laryngocarcinoma and nasopharyngeal carcinoma / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To explore the significance of PML expression in carcinogenesis of laryngocarcinoma and nasopharyngeal carcinoma.@*METHOD@#The PML expression in 56 laryngocarcinoma tissues and 34 nasopharyngeal carcinoma tissues were examined by immunohistochemistry. Moreover, the relationship between the PML expression, with the pathologic grade,T stage and lymph node metastasis was analyzed.@*RESULT@#The PML expression decreased with the ascending of pathological tumor grade (P < 0.05) and with the increasing of T grade of tumor (P < 0.05). The expression of PML in nasopharyngeal carcinoma was lower than that in carcinoma of larynx (P < 0.05). However, either in laryngocarcinoma or in nasopharyngeal carcinoma, there was no statistical difference on PML expression between with and without lymph node metastasis in laryngo carcinoma tissues and nasopharyngeal carcinoma tissues.@*CONCLUSION@#The close relationship between PML expression and the malignant parameters such as pathological grade and T grade of laryngocarcinoma and nasopharyngeal carcinoma, indicated that PML may play a role in cancer repression but not in cancer metastasis.

The changes of GABA and GABAergic neurons in inferior colliculus of unilateral cochlear damage rats / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To observe the changes of GABA and GABAergic neurons in rat inferior colliculus following unilateral cochlear damage and explore the function and significance of GABA in reorganization of auditory center after deafferentation.@*METHOD@#Twenty sprague dawley rats were divided into four groups randomly. The technique of direct anti-GABA immunocytochemistry (SP) was used in this study. The quantity of GABA was measured by 835-50 type Amino Acid Automatism Analyzer and compared at one week, two weeks and one month after unilateral cochlear ablation, respectively.@*RESULT@#Compared with controlled groups, the quantity levels of GABA decreased and the number of GABAergic neurons also reduced one week after unilateral cochlear damage and there was a significant difference in GABA between two groups( P <0. 05); after one month, the quantity of GABA and the number of GABAergic neurons almost reached the normal level, the differences were no significant.@*CONCLUSION@#The dynamic change of GABA in rat inferior colliculus reflected the neuronal activity, which implied that GABA may play an important role in reorganization of auditory center after unilateral cochlear damage.

Expression of RAD51 in carcinomas of cancer of larynx and its clinical significance / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To investigate the expression and clinical significance of RAD51 in laryngocarcinoma.@*METHOD@#The expression of RAD51 in laryngocarcinoma and polyp of vocal cord tissues were determined by immunohistochemical staining. The results were analyzed and compared with the clinical stage, lymph node metastasis and pathologic grade.@*RESULT@#(1) The expression of RAD51 in laryngocarcinoma group was extremely stronger than that in polyp of vocal cord group (P < 0.01). (2) There was significant difference for RAD51 expression in cancer cells between earlier clinical stage group and advanced clinical stage group (P < 0.01). (3) There was significant difference for RAD51 expression in different pathologic grades (P < 0.05). (4) There was also significant difference for RAD51 expression between groups with and without lymph node metastasis.@*CONCLUSION@#RAD51 may play a critical role in tumorigenesis of laryngocarcinoma. RAD51 may be a potential marker for clinical diagnosis and treatment of Laryngocarcinoma. It may be significant in predicting clinical stage, pathologic grade and metastasis.

Expression of human protection of telomeres1 (POT1) and telomeric repeat binding factor-2 (TRF2)in laryngneal squamous cell carcinoma / 中国耳鼻咽喉头颈外科

OBJECTIVE To study the significance by analyzing the expression of human protection of telomeres1(POT1)and telomeric repeatbinding factor-2(TRF2)in the tissues of laryngneal squamous cell carcinoma(LSCC)and in polyp of vocal cord tissues. METHODS The expression of POT1 and TRF2 in 20 tumor samples and 19 polyp of vocal cord were determined by the indirect immunofluorescence method. The results were scored and analyzed statistically. RESULTS The positive expression rates of POT1 and TRF2 in tumor samples was 65.00% and 70.00% respectively. There were no positive expression of POT1 and TRF2 in polyp of vocal cord. The positive expression of POT1 was higher in poorly differentiated laryngeal carcinoma than that in well-differentiated and moderately differentiated laryngeal carcinoma(Chi-square test with contingency table, P0.05). CONCLUSION The expression of POT1 and TRF2 in laryngneal squamous cell carcinoma were remarkable, POT1 and TRF2 may play a critical role in tumorigenesis of laryngneal squamous cell carcinoma. There was statistical significant difference for degrees of POT1 expression in different tumor histological grades.

Influence of telomerase reverse transcriptase shRNA on expression of c-myc in nasopharyngea carcinoma cells / 中国耳鼻咽喉头颈外科

OBJECTIVE To investigate the influence of short hairpin RNA targeting human telomerase reverse transcriptase(hTERT)on inhibition of telomerase activity and on expression of protein c-myc in nasopharyngeal carcinoma cells. METHODS Plasmid shRNA1 containing fluorescein gene and hTERT cDNA sequences were synthesized. Cells were transfected with plasmid shRNA1. The cell viability was examined using the MTT assay. The activity of telomerase was tested by polymerase chain reaction telomeric repeat amplification protocol- enzyme-linked immunosorbent assay (PCR-TRAP- ELISA),protein c-myc expression was tested by western blot. RESULTS It was observed that treatment with pshRNA1 in the presence of a valid transfection reagent could significantly reduce telomerase activity and the expression of protein of c-myc. CONCLUSION Inhibition of telomerase activity or expression of hTERT mRNA in nasopharyngeal carcinoma cells could inhibit cells proliferation and reduce the expression of protein of c-myc.

The changes of GABA and Glu in rat inferior colliculus after unilateral cochlear ablation / 中国耳鼻咽喉头颈外科

OBJECTIVE To observe the changes of GABA and Glu in rat inferior colliculus following unilateral cochlear ablation and explore the function and significance of GABA and Glu in reorganization of auditory center after deafferentation. METHODS Twenty-five Sprangue Dawley (SD) rats were divided randomly into 5 groups. The content of GABA and Glu were measured by 835-50 type Amino Acid Automatism Analyzer and compared at 1 week, 2 week and 1 month after unilateral cochlear ablation respectively. RESULTS Compared with sham operated groups, the content levels of GABA decreased (from 78.00?7.50 to 51.65?10.36, about decreasing 33.6 %)1 week after unilateral cochlear ablation and there was a significant difference in GABA levels between 2 groups(P0.05). CONCLUSION The dynamic change of GABA and Glu in rat inferior colliculus reflected the neuronal activity, which implied both GABA and Glu may play an important role in reorganization of auditory center after unilateral cochlea ablation.

Research on DPOAE of guinea pigs under awake and anesthetic conditions / 生物医学工程学杂志

In order to observe the DPOAE of guinea pigs under awake and anesthetic conditions, we invesitigated the amplitude of distortion product emission and I/O function curves and made a comparative analysis of their changes under different conditions. It was found that the amplitude of DPOAE and I/O curves did not show difference in the guinea pigs under awake and anesthetics conditions. We deem it credible and simple to measure DPOAE in the guinea pigs under awake condition, which is similar to measuring DPOAE in the guinea pigs under physiological condition.