Study on interference therapy induced by epidermal growth factor receptor-antisense cDNA in signal transduction of laryngeal squamous cell carcinoma / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To investigate effect of interference therapy induced by epidermal growth factor receptor (EGFR)-antisense cDNA in signal transduction of Hep-2 laryngeal squamous cell carcinoma in vitro.</p><p><b>METHODS</b>AdEasy Vector System was used to construct the recombinant adenovirus vector sense/antisense-pAdEasy-EGFR. The recombinant adenovirus vector introduced EGFR-sense/antisense cDNA fragment into HEK293 cell. The purified recombinant adenovirus sense/antisense-pAdEasy-EGFR transfected Hep-2 cells in vitro. The inhibition of EGFR protein expression and proliferation of Hep-2 cells, the changes of cell cycle and DNA content in Hep-2 cells were examined by MTT, Western blot analysis, flow cytometry essay, and immunocytochemistry respectively.</p><p><b>RESULTS</b>The higher titre sense and antisense mRNA expression recombinant adenovirus containing 1,032 bp EGFR-cDNA was constructed and prepared successfully. When antisense-pAdEasy-EGFR was transferred into Hep-2 cells the inhibition of cell proliferation and EGFR protein expression in Hep-2 cells were investigated effectively.</p><p><b>CONCLUSION</b>The antisense-pAdEasy-EGFR effectively interfere the Hep-2 signal transduction pathway and induce apoptosis which results in inhibiting proliferation of laryngeal squamous cell carcinoma.</p>

Construction of recombinant adenovirus vector carrying cell cycle controlling gene-p14ARF / 华西口腔医学杂志

<p><b>OBJECTIVE</b>The recombinant adenovirus vector carrying p14ARF gene was constructed for using in the interference therapy in signal transduction of laryngeal squamous cell carcinoma.</p><p><b>METHODS</b>The total cDNA fragment of p14ARF was cloned into the shuttle plasmid pAdTrack-CMV, with the resultant plasmid and the backbone plasmid pAdEasy-1, the homologous recombination took place in the E.Coli BJ5183 and the recombinant adenoviral plasmid was generated. The adenoviruses were packaged and amplified in the 293 cells. Then the viral titer was checked by GFP.</p><p><b>RESULTS</b>The recombinant adenovirus vector carrying p14ARF was constructed successfully. The viral titer was 2.3 x 10(9).</p><p><b>CONCLUSION</b>The recombinant adenovirus vector could introduce p14ARF gene into the laryngeal squamous cell carcinoma line or tumor tissue effectively, which would provide experimental basis for the mechanisms and further study of the interference therapy in signal transduction of laryngeal squamous cell carcinoma.</p>

Clinical anatomy measurement of accessory nerve in neck dissection / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To investigate the relations between accessory nerve and its surrounding structures.</p><p><b>METHODS</b>One hundred and thirty six patients were divided into two groups: has or has no neck surgical history. Neck dissection were performed and the four distance were measured simultaneously. The distance of accessory nerve and the great auricular nerve going out the posterior edge of sternocleidomastoid muscle; the distance of the point accessory nerve going out the posterior edge of sternocleidomastoid muscle to clavicular midpoint; the distance of the point accessory nerve going out the posterior edge of sternocleidomastoid muscle to sternoclavicular articulation; the distance of the point accessory nerve enter trapezius muscle to clavicular midpoint.</p><p><b>RESULTS</b>In no neck dissection group, the point accessory nerve going out sternomastoid muscle were supra the point of great auricular nerve going out the sternomastoid muscle, the average length of two points is (0. 61 +/- 0. 35) cm , the significance has not observed between genders (P > 0.05), however, there has significant difference between two groups of has or has no neck surgical history (P < 0.05). 88.2% (112/127) accessory nerve going out supra the great auricular within 1.0 cm, 11.8% (15/127) within 1.0 approximately 2.0 cm. 67.7% (86/127) accessory nerve adopt branch from cervical plexus before entering trapezius. The distances of the point accessory nerve going out the posterior edge of sternocleidomastoid muscle to clavicular midpoint and to sternoclavicular articulation were significant relative not with before neck surgical history but gender. The distance of the point accessory nerve enter trapezius muscle to clavicular midpoint is (4.96 +/- 0.78) cm, it has no difference both before neck surgical history and gender (P > 0.05).</p><p><b>CONCLUSION</b>In no neck surgical history group,both of the distance that accessory nerve and the great auricular nerve going out the posterior edge of sternocleidomastoid muscle and the point accessory nerve enter trapezius muscle to clavicular midpoint were helpful for search accessory nerve in surgery. But in patients who have neck surgical history or great auricular have been injured, accessory nerve could be looked for associating with the distances of the point accessory nerve going out the posterior edge of sternocleidomastoid muscle to clavicular midpoint and to sternoclavicular articulation; the distance of the point accessory nerve enter trapezius muscle to clavicular midpoint.</p>

Specific inhibition of Raf-1 gene expression in HNE1 by RNA interference / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To silence the expression of Raf-1 gene in HNE1 cells using vector-based RNA interference (RNAi) technique.</p><p><b>METHODS</b>The vector containing the human U6 promoter was used for targeted gene silencing when a dsDNA oligonucleotide encoding an appropriate shRNA was ligated into the vector, and 67nt oligonucleotide fragment was inserted into the downstream of the U6 promoter. Plasmids containing different Raf-1 target sequences [ (1) pshuttle-Raf-1-a( 225), (2) pshattle-Raf-1-b ( 358) and (3) pshuttle-Raf-1-c(474)], were transfected into HNE1 cells. Expression of Raf-1 mRNA was assayed by RT-PCR. Apoptosis were determined by cytometry.</p><p><b>RESULTS</b>Vector-based RNAi had advantages over antisense RNA because it could be delivered to the target cell more efficiently, and effect could last longer. Raf-1 expression could be inhibited by plasmid-expressed shRNA. Three different targeting sequences were selected from Raf-1 gene, and the inhibitory effect of pSIREN shuttle-Raf-1-b (358) was biggest.</p><p><b>CONCLUSION</b>Raf-1 expression in HNE1 cells can be inhibited significantly using plasmid-based RNAi.</p>

Microsurgical anatomy of epiglottic artery and its clinical implications / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To study the microsurgical anatomy of human epiglottic blood vessel to provide exact and reliable data and morphological properties for further studies of laryngeal transplantation, reconstruction and other epiglottis-related diseases.</p><p><b>METHODS</b>Thirty Chinese adult cadavers (27 males and 3 females) were examined for the present study. The cadavers were dissected under magnification along the anatomic planes from skin down to vertebral column. The anterior neck was exposed by a midline incision and extended laterally along the angle of mandible superiorly, and on clavicle inferiorly. After the visualization of laryngeal prominence of thyroid cartilage, strap muscles were resected and superior laryngeal artery and epiglottic blood vessel were exposed under an operating microscope ( original magnification 7 -30). The epiglottic artery was named for the first time.</p><p><b>RESULTS</b>The diameter of superior laryngeal artery was (1. 06+/-0. 16) mm( male: 1. 09 mm+/-0. 12 mm). The diameter of origin epiglottic artery was (0. 79+/-0. 13) mm (male: 0. 81 mm+/-0. 11 mm). The vertical distance between origin epiglottic artery and superior horn of thyroid cartilage was (27. 16+/-3. 85) mm. Epiglottic artery loop was observed in all the cadavers, which could be M-, N-, omega-, or U-shaped and mixed under the thyrohyoid membrane or in the epiglottic vallecula.</p><p><b>CONCLUSIONS</b>These findings could improve the application of epiglottis in laryngeal functional reconstruction after partial laryngectomy, as well as in the prevention of epiglottic artery loop injuries during the operation.</p>

Anatomic study of pterygopalatine fossa under endoscope / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To provide endoscopic anatomic bony structures of pterygopalatine fossa for skull base surgery.</p><p><b>METHODS</b>The bony structures of the pterygopalatine fossa were observed in ten dry skulls under endoscope.</p><p><b>RESULTS</b>The pterygopalatine fossa showed a long and narrow cleft composed of the body and pterygoid process of sphenoid bone, the lamina perpendicular of palatine bone, and the posterior wall of maxillary sinus. The pterygopalatine fossa is (21.4 +/- 0.8) mm x (5.2 +/- 0.3) mm x (3.2 +/- 0.3) mm, with seven paths communicating with nasal cavity, mouth cavity, pharynx, orbit, infratemporal fossa and middle cranial fossa. Under endoscope,the whole pterygopalatine fossa could be observed.</p><p><b>CONCLUSIONS</b>Endoscopic anatomic study of the pterygopalatine fossa is important to endoscopic endonasal skull base surgery. Under endoscope,the whole pterygopalatine fossa can be observed.</p>

Establishment of a modified intranasally ovalbumin induced animal model of allergic rhinitis / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To observe the early and late symptomatic, pathological and immunological changes in an intranasal ovalbumin-induced animal model of allergic rhinitis in guinea pigs.</p><p><b>METHODS</b>Guinea pigs were intranasally sensitized with ovalbumin absorbed on aluminum hydroxide and after 5 days' interval, they were challenged with 1% ovalbumin solution once every 3 days for total 11 times. Two control groups were studied in parallel, the positive treatment control group was treated with antihistamine and the negative control group was sham-sensitized and sham-challenged. Typical symptoms of allergic rhinitis, such as sneezing, nasal scratching, nasal blockage and rhinorrhea were evaluated. Passive cutaneous anaphylaxis reaction (PCA) was performed to measure the levels of IgG1 and IgE. Eosinophils infiltration and goblet cells in nasal mucosa were observed. In addition, the level of histamine and the number of total leukocytes and eosinophils in the nasal lavage fluid were also measured.</p><p><b>RESULTS</b>In the model group, symptoms of sneezing, nasal scratching, nasal blockage and rhinorrhea were induced after ovalbumin challenge. The respiratory rate (RR), which reflected the resistance of upper airway, showed a biphasic change. In the PCA test, IgG1 and IgE levels increased after challenges. Eosinophil infiltration in nasal mucosa was more obvious in active groups in comparison to with the negative control group (P < 0.05 or < 0.01). The histamine, total leucocytes and eosinophils levels in nasal lavage fluid also showed higher in the model group (P < 0.05 or < 0.01). The antihistamine treated animals were also induced out above changes but modest compared with the model group (P < 0.05 or < 0.01). The negative control showed few of above changes with significant difference (P < 0.05 or < 0.01).</p><p><b>CONCLUSIONS</b>Our results implied that the modified animal model of allergic rhinitis was capable of showing satisfactory symptomatic and pathophysiological changes in allergic rhinitis. It showed a biphasic nasal blockage with shorter establishment duration. The model also had good treatment reaction to antihistamine. The animal model we introduced may be useful in the study of allergic rhinitis.</p>

Effects of budesonide, desloratadine and dexamethasone on interleukine-4 release and expression from human mast cell line / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>Since human mast cell is an important source of cytokines, it is of importance to understand the effects of anti-allergic drugs on cytokines modulation in mast cells. In the present study, we aimed at observing whether IL-4 could be released from human mast cell line (HMC-1) after the stimulation of PMA + A23187, and the effects of systemic glucocorticosteroid, dexamethasone, topical glucocorticosteroid, budesonide and H1 antagonist, desloratadine on IL-4 release and mRNA expression.</p><p><b>METHODS</b>HMC-1 was stimulated with 25 ng/ml phorbol 12-myristate 13-acetate (PMA) and 2.5 x 10(-7) mol/L ionomycin (A23187) and cultured for 6 hours, 12 hours and 24 hours respectively in the presence or absence of 10(-6)-10(-10) mol/L concentrations of test drugs. Culture supernatants were collected and the levels of IL-4 were assayed by enzyme-linked immunosorbent assays (ELISA). The mRNA expression of IL-4 was measured by semiquantitative reverse transcription-polymerase chain reaction (RT-PCR).</p><p><b>RESULTS</b>HMC-1 expressed IL-4 mRNA and the resulting protein production of IL-4 released after being stimulated with PMA plus A23187. Dexamethasone, budesonide and desloratadine had potent inhibitory effect on IL-4 release at any concentrations and time points, with significant deference (P < 0.05) compared to the control cells. The inhibitory effect did not show time-dependent and concentration-dependent manner. Desloratadine and budesonide showed neither up-regulatory nor down-regulatory effects on IL-4 mRNA expression at the test concentrations, however, desloratadine could down-regulate IL-4 mRNA expression.</p><p><b>CONCLUSIONS</b>HMC-1 could express and produce IL4 after stimulation. Dexamethasone, budesonide and desloratadine all had inhibitory effects on IL-4 release from HMC-1. In addition, desloratadine could also inhibit the IL-4 mRNA expression.</p>

Measurement of sound pressure level at outlet of external auditory canal and eardrum / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To evaluate the sound pressure level at outlet of external auditory canal and eardrum, and to observe the range of maximum frequency of sound effect of outlet ear canal, so as to provide exact data for designing personal noise rating instrument and studying the effect of environmental noise on human body, and to provide basis for calibration of noise sound pressure level.</p><p><b>METHOD</b>Sound pressure level at eardrum and outlet of external auditory canal in 30 young people (60 ears) were measured by Audio Scan RM500 at 50 dB SPL narrow band noise with different frequencies (0.25 - 8.00 kHz).</p><p><b>RESULTS</b>At the same frequency, sound pressure levels at eardrum were less than (0.6 +/- 2.6) dB SPL, at outlet of outlet ear canal less than (0.2 +/- 1.8) dB SPL, there were no significant differences between both sound pressure levels (P > 0.05). There were significant differences in sound pressure levels between both places at 1.50 - 8.00 kHz (P < 0.01). The maximum difference was (10.5 +/- 3.4) dB SPL at 2.00 kHz, and the minimum was (0.5 +/- 6.2) dB SPL at 0.50 kHz.</p><p><b>CONCLUSION</b>The effects of increased sound pressure should be considered in the evaluation of environmental noise and the design for individual noise rating meter when the noise frequency was >or= 1.50 kHz.</p>