Study on expression of IL-13 and correlation between IL-13 and MUC5AC in eosinophilic chronic rhinosinusitis with nasal polyps / 中国耳鼻咽喉头颈外科

OBJECTIVE To explore expression of IL-13 in eosinophilic chronic rhinosinusitis with nasal polyps(EOS-CRSwNP) and further investigate the correlation between IL-13 and MUC5AC in EOS-CRSwNP. METHODS MUC5AC was detected in tissues from normal nasal mucosa and EOS-CRSwNP by immunohistochemistry and ELISA. We also used ELISA to detect the expression of IL-13 in normal nasal mucosa and EOS-CRSwNP. Correlation between IL-13 and MUC5AC in EOS-CRSwNP was investigated by correlation analysis. Secretion of MUC5AC in IL-13 incubated primary air-liquid interface(ALI)-cultured nasal polyp epithelial cells was examined by ELISA. RESULTS MUC5AC is mainly expressed in the epithelium of nasal mucosa by immunohistochemistry. By ELISA, expressions of MUC5AC and IL-13 are higher in EOS-CRSwNP than those in normal nasal mucosa. Correlation analysis shows that there exists a high correlation between MUC5AC and IL-13 in EOS-CRSwNP. IL-13 upregulates expression of MUC5AC in IL-13 incubated primary ALI-cultured nasal polyp epithelial cells for 7 or 14 days. CONCLUSION Expressions of MUC5AC and IL-13 are higher in EOS-CRSwNP than those in normal nasal mucosa respectively, and MUC5AC correlates with IL-13 highly in EOS-CRSwNP.

Allergen-Dependent Differences in ILC2s Frequencies in Patients With Allergic Rhinitis

PURPOSE: Group 2 innate lymphoid cells (ILC2s) are a novel population of lineage-negative cells that induce innate type 2 responses by producing the critical Th2-type cytokines IL-5 and IL-13 in response to IL-25 and IL-33 stimulation. ILC2s accumulation in the peripheral blood of patients with allergic rhinitis (AR) is controversial; the precise role of ILC2s in the immunopathogenesis of AR is still not clear. We investigated the role of ILC2s in phenotypic AR sensitized to distinct allergens. METHODS: Flow cytometric analysis of the peripheral blood of 7 healthy controls (HCs), 9 patients monosensitized to house dust mite (HDM), and 8 patients monosensitized to mugwort was performed to quantify ILC2s frequency. Peripheral blood mononuclear cells (PBMCs) were isolated from HDM-AR and mugwort-AR patients, and Lineage- and Lineage+ cells were separated using a fluorescence-activated cell sorter (FACS). IL-5 and IL-13 levels in the supernatants of PBMCs, and Lineage- and Lineage+ cells stimulated with IL-25 and/or IL-33 combined with IL-2 in vitro were assessed using the Milliplex magnetic bead kit. RESULTS: The percentage of ILC2s was significantly elevated in HDM-AR patients compared to mugwort-AR patients and HCs, while no significant difference was found between mugwort-AR patients and HCs. IL-33+/-IL-25 plus IL-2 induced a significantly greater release of IL-5 and IL-13 in the PBMCs of HDM-AR patients compared to PBMCs of mugwort-AR patients. IL-25 plus IL-2 also induced a significantly greater release of IL-13 in the PBMCs of HDM-AR patients compared to PBMCs of mugwort-AR patients. Stimulation with IL-33 and/or IL-25 combined with IL-2 also induced a significantly greater IL-5 and IL-13 release from Lineage- cells compared to Lineage+ cells. CONCLUSIONS: AR patients sensitized to HDM or mugwort allergen have distinct phenotypic and functional profiles in ILC2s frequencies. ILC2s mediate major type 2 immunity in the development of HDM-AR and may be a potential therapeutic target.

TMEM16A-Mediated Mucin Secretion in IL-13-Induced Nasal Epithelial Cells From Chronic Rhinosinusitis Patients

PURPOSE: Chronic rhinosinusitis with nasal polyps (CRSwNP), a mainly Th2 cytokine-mediated disease, often involves mucus secretion. Recent evidence suggests that transmembrane protein 16A (TMEM16A), a calcium-activated Cl- channel (CaCC), can regulate mucus secretion from airway epithelium by transepithelial electrolyte transport and hydration. However, the role of TMEM16A in mucin production/secretion in the airway epithelium is not clear. This study was conducted to determine the role of TMEM16A in mediating mucin secretion in human nasal polyp epithelial cells (HNPECs) induced by IL-13. METHODS: Human sinonasal mucosa tissue and dissociated sinonasal epithelium from control subjects and patients with CRSwNP were assessed for the expression of TMEM16A and the secretion of human mucin 5AC (MUC5AC) by immunohistochemistry, Western blot analysis, and enzyme-linked immuno-sorbent assay (ELISA). A model of the Th2 inflammatory environment was created by exposure of primary air-liquid interface (ALI)-cultured HNPECs to interleukin-13 (IL-13) for 14 days, with subsequent assessment of TMEM16A expression in cell lysates by Western blotting and MUC5AC secretion in apical washings of cells by ELISA. RESULTS: The expressions of TMEM16A and MUC5AC were increased in human nasal polyp tissue and dissociated nasal polyp epithelium. TMEM16A was detected in IL-13-treated HNPECs, specifically in MUC5AC-positive cells but not in ciliated cells. IL-13 treatment increased percentages of TMEM16A-positive cells, MUC5AC-positive cells, and cells coexpressing TMEM16A/MUC5AC, the expression of TMEM16A protein, and the secretion of MUC5AC. T16Ainh-A01, a TMEM16A inhibitor, attenuated these IL-13-induced effects. CONCLUSIONS: The expression of TMEM16A and MUC5AC are increased in CRSwNP, which might be a direct effect of Th2 cytokines present in the sinonasal mucosa in CRSwNP. Down-regulation of TMEM16A expression and MUC5AC secretion in HNPECs by T16Ainh-A01 indicates that TMEM16A might play an important role in mucin secretion in upper airway inflammatory diseases.

Onset feature and efficacy of early interventional treatment of Artemisia pollinosis / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To analyze the clinical feature and treatment methods of Artemisia pollinosis.</p><p><b>METHODS</b>Skin prick test results of 14 426 cases from Beijing Tongren hospital and pollen concentration of Beijing observatory from 2007 to 2011 were analyzed to identify the clinical feature of Artemisia pollinosis patients and its correlation with the pollen concentration. Patients were given leukotriene receptor antagonists (Montelukast) for 2 weeks, followed by 4 weeks of mometasone furoate nasal spray (EIT group: n = 21), or only 4 weeks of mometasone furoate nasal spray (POT group: n = 16). The nasal symptom score was compared between 2 groups.SPSS 16.0 software was used to analyze the data.</p><p><b>RESULTS</b>Artemisia pollinosis accounted for 30.8% (4 442/14 426) of all SPT positive allergic rhinitis patients, and most Artemisia SPT positive results were strong positive(3 793/4 442, 85.4%); onset age peak of Artemisia pollinosis patients was at the age of 19 to 30, onset time concentrated in August to September, was consistent with the peak period of Artemisia pollen concentration; EIT treatment using leukotriene receptor antagonists two weeks before pollen season significantly improved sneeze, sniveling and rhinocnesmus symptoms (t value was 3.28, 3.92, 3.09, respectively, all P < 0.01) compared with post-onset treatment (POT). But nasal obstruction and cough symptoms had no significant difference between two groups (t value was 0.85, 1.52, respectively, all P > 0.05).</p><p><b>CONCLUSION</b>Artemisia pollen is the main pollen allergen in Beijing, EIT treatment was effective to pollinosis.</p>

The Effects of interleukin-18 on Middle Ear Inflammation in the Rat Model of Otitis Media with Effusion / 听力学及言语疾病杂志

Objective To investigated the effects of IL -18 on middle ear allergic inflammation in rat model of OME .Methods Eighteen SD rats were randomly divided into three group :group A(control group ,n=12 ears) , group B (OME model group ,n=12 ears) ,group C (IL -18 injectiion group ,n=12 ears) .The rat model of OME was established by sensitizing with ovalbumin (OVA) and later challenging in tympanic bullae .Recombinant rat IL-18 (1 μg ,+0 .2 ml saline ,) were injected in group C at 1 ,2 ,7 ,8 ,15 ,16 day .At the same time sacle ,0 .2 ml sa-line ,instead of IL -18 ,were intraperitoneal injection in group A and B .The morphologic changes of the middle ear epithelial cells and inflammatory cells infiltration were observed under light microscope .The level of IFN -γand IL-4 in tympanic lavage fluid(TLF) were determined by ELISA .Results Pathological examination showed that middle ear mucosa inflammation and eosinophil infiltration in group C were no less severe than group B .The numbers of neutrophils in group C increased significantly compraring with group B (P0 .05) ,while significantly greater than that in group A (P0 .05) .Addtionally ,there was no significant difference in the ratio of Th1 /Th2(IFN -γ/IL -4)between group B and group C (P>0 .05) . Conclusion IL -18 acts as an immune regulatory factors ,significantly increases Th1 cytokine IFN -γ.Although to some extent alleviate the OME rat middle ear Th1 /Th2 imbalance ,there is still excessive activation of Th cells . Th1 and Th2 cells factor are excessive for the secretion disorder of the immune response status .The OME rat mid-dle ear allergic inflammation has not been fundamentally alleviated ,the underlying mechamism should be further studied .

The observation of the inflamed cells in the mucosa at the medial site compare with that at the lateral site of the normal uncinate process / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To determine the inflammatory cells in the mucosa at the medial aspect of the normal uncinate process compared with that on the protected lateral aspect of the normal uncinate process.@*METHOD@#The mucosa of 20 uncinate process from the nasal cavity of 17 patients with no evidence of sinus disease undergoing functional endoscopic sinus surgery were recruited for the study. The material was stained with HE, Chromotrope 2R, Alcian blue-periodic acid-schiff, Toluidine blue. Specimens were observed using an Olympus microscope.@*RESULT@#The number of mast cells and goblet cells were found to be higher on the lateral aspect of the normal uncinate process than on the medial aspect. The number of plasma cells was obviously different from that of lymphocytes. We did not found any eosinophils on either sides of uncinate process.@*CONCLUSION@#There are differences in the number of mast cells and goblet cells between the mucosa at the medial aspect of the normal uncinate process and the mucosa at the protected lateral aspect of the normal uncinate process.

Characteristics of intrinsic laryngeal muscle after recurrent laryngeal nerve injury / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To investigate the electrophysiological characteristics and pathological changes in different recurrent laryngeal nerve injury models.@*METHOD@#Twenty dogs were divided into different models of recurrent laryngeal nerve injury. Electrophysiological changes of animal models and histopathological changes in laryngeal muscles were investigated at different time.@*RESULT@#Amplitude of MUP in TA decreased at first and then increased a little, while duration of MUP keeps decreased. The latent periods of the evoked potential in the incomplete injury group increased at first and then went down. In animal histopathological investigation, the lighter of the muscle injuries and the bigger of the diameter of muscle fibers and fascicles, and the smaller of the number of cellular nucleolus per square inch. In the early periods after injury, the correlation among the standard amplitude, threshold amplitude, maximum amplitude of EP and the diameter of fascicles were positive; while in the late period, the correlation among the amplitude, duration, area of MUP and nucleolus number was positive.@*CONCLUSION@#The physiological characteristics of laryngeal nerves and muscles could be known. LEMG was an important method in diagnosis of laryngeal nerve and muscle diseases.

The changes of phosphorylated c-Jun expression in spiral ganglion after exposed to noise / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To investigate the mechanism of hearing pathway NMDAR regulating the changes of phosphorylated c-Jun expression in spiral ganglion after exposed Kunming mice to neural injury noise stimulation to induce permanent or temporary threshold shift.@*METHOD@#To compare the different expressions of the key components (phosphorylated C-Jun) of NMDAR signal pathway during neural injury stimulation by Immunohistochemistry in CG.@*RESULT@#The levels of phosphorylated c-Jun remarkably, increased in the spiral ganglion after 8 h, 48 h, 7 d and 14 d following noise trauma induced permanent threshold shift (PTS), and the numbers of positive cells reduced gradually. The similar changes occur in mice treated with MK-801 30 minutes before and after 3 h trauma induced PTS. After 48 h of noise induced TTS, the expression of Phosphorylated c-Jun return the level of normal control.@*CONCLUSION@#The expressions of phosphorylated c-Jun are time-related and uniform in the time and position in CG after noise trauma. MK-801 can alleviate the damage of noise trauma by altered the NMDA receptor-mediated calcium influx. Therefore, the NMDA receptors may involved in the damage of inner ear in common.

Expression of NR2A in rat auditory cortex after sound insulation and auditory plasticity / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To study the changes of N-methyl-D-aspartate (NMDA) receptor subunit 2A (NR2A) expression at local synapses in auditory cortices after early postnatal sound insulation and tone exposure.@*METHOD@#We prepared highly purified synaptosomes from primary auditory cortex by Optiprep flotation gradient centrifugations, and compared the differences of NR2A expression in sound insulation PND14, PND28, PND42 and Tone exposure after sound insulation for 7 days by Western blotting.@*RESULT@#The results showed that the NR2A protein expression of PND14 and PND28 decreased significantly (P0.05).@*CONCLUSION@#The results indicate that sound insulation and experience can modify the protein expression level of NR2A during the critical period of rat postnatal development. These findings provide important data for the study on the mechanisms of the developmental plasticity of sensory functions.

The expression and significance of NF-kappaB in the rat model of otitis media with effusion experimentally induced by ovalbumin / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To investigate the T helper cells (Th) predominant differentiation and the modulation of nuclear transcription factors kappa B (NF-kappaB) in middle ear of rat model of otitis media with effusion (OME).@*METHOD@#Sixteen SD rats were randomly divided into OME (Exp group) and control group (Con group). The expression of NF-kappaB were observed by immunohistochemistry. The level of IFN-gamma and IL-4 in tympanic lavage fluid (TLF) were determined by ELISA.@*RESULT@#As compared to the Con group , the level of IL-4 and the ratio of Th2/Th1 (IL-4/IFN-gamma) in TLF of Exp group significantly increased (P<0.05), when no significant difference in IFN-gamma levels in TLF was found. The ratio of NF-kappaB p65 positive cells to white cells in temporal bone marrow smears and middle ear mucosa of Exp group was significantly higher than that of Con group (P<0.05). The expression of NF-kappaB p65 in temporal bone marrow smears and middle ear mucosa was signficantly positively correlated with the concentration of IL-4 in TLF of Exp group (P<0.05).@*CONCLUSION@#The middle ear is capable of mounting an allergic response and subsequent formation of effusion. There is Thl/Th2 immune response imbalance, which polarizes toward Th2 response in the middle ear microenvironment of allergic OME rat model. Moreover , NF-kappaB may participate in regulating Th2 predominant reaction.

Expression and significance of TGF-beta1 and HSP70 in human laryngeal squamous cell carcinoma / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To investigate the expression of TGF-beta1 and HSP70 in human laryngeal squamous cell carcinoma.@*METHOD@#The expression levels of TGF-beta1 and HSP70 in 53 specimens of human laryngeal squamous cell carcinoma and 48 specimens of para-carcinoma were detected by immuno- histochemistry and computer assisted image analysis.@*RESULT@#The expression of TGF-beta1 was decreased in carcinoma tissues compared with para-carcinoma tissues ( P < 0.05), while the expression of HSP70 was increased ( P < 0.05). Both the expression of TGF-beta1 and HSP70 were significantly correlated with the differentiation of laryngeal squamous cell carcinoma (P < 0.01 or P < 0.05). However, there was a negative correlation between TGF-beta1 and HSP70 (r = -0.87, P < 0.01).@*CONCLUSION@#TGF-beta1 and HSP70 play an important role in malignant behaviors of human laryngeal carcinoma.

Microsatellite instability and abnormal mismatch repair in laryngeal squamous cell carcinoma / 中国耳鼻咽喉头颈外科

OBJECTIVE The aim of our study was to evaluate the significance of the frequency of microsatellite instability (MSI) and its relationship to mismatch repair gene (MMR) in laryngeal squamous cell carcinoma.METHODS We investigated the expression frequency and clinical significance of MSI and MMR in 50 laryngeal squamous cell carcinoma (LSCC) patients from Beijing Tongren Hospital. The status of MSI was evaluated using microdissection - polymerase chain reaction (PCR) - single strand length polymorphism (SSLP) - silver staining.Five markers on chromosomes 1p, 3p, 5q, 9p, 17p, which were adjacent to BCAR3 (breast cancer anti-estrogen resistance 3), FHIT, APC, CDKN2A (p16), TP53 respectively, were used. Two of the six components of MMR -hMLH1 and hMSH2- were investigated by an immunohistochemical approach because of the high frequency of their downregulation in head and neck tumors. RESULTS The informative case number of the five markers (D17S796, D3S3544, D5S656, D1S375, D9S162) were 44, 42, 45, 44 and 40 respectively. The incidence of MSI was lower than the frequency of loss of heterozygosity (LOH). The incidence of MSI on D17S796 (TP53)was 20.5% (9/44),on D3S3544 (FHIT) was 14.3 % (6/42), on D5S656 (APC) was 31.1% (14/45), on D1S375 (BCAR3) was 20.5 % (9/44), and on D9S162 (CDKN2A) was 15.0 % (6/40). Though there was no relationship between MSI status and age, gender, smoke history, tumor location, tumor differentiation and T stage (P＞0.05),there was a strong correlation between MSI and relapse condition (P＜0.01). Also, MSI status correlated with MMR expression to some degree (P＜0.01), but it was common for negative and positive staining of MMR to coexist on the same slide. CONCLUSION Microsatellite instability and abnormal mismatch repair may contribute to the carcinogenesis of a subset of laryngeal squamous cell carcinoma. Microsatellite instability may be a characteristic signal of tumor recurrence.

Expression of interleukin -12 and correlation between interleukin -12 and eosinophils in nasal mucosa in allergic rhinitis mice / 中国耳鼻咽喉头颈外科

OBJECTIVE To observe the expres-sion of interleukin-12 and eosinophils in the nasal mu-cosa of allergic rhinitis mice. METHODS Thirty nine male BALB/c mice,6～8 weeks old,were randomly divided into three groups: control group,allergic rhinitis (AR)group, and Budesonide treatment group. Al-lergic rhinitis model in mice were established by using ovalbumin intraperitoneal immunization and nasal anti-gen challenge. The nasal mucosa obtained from mice of three groups were stained routinely by HE and im-munohistochemical method to observe the distribu-tion and expression of interleukin-12 and eosinophils. RESULTS The expression of eosinophils in the nasal mucosa of AR group was significantly higher than con-trol group(P

Detection of disseminated tumor cells in the peripheral blood of patients with laryngeal or laryngopharyngeal carcinomas / 中国耳鼻咽喉头颈外科

OBJECTIVE To investigate the diagnostic and prognostic value of molecular biological detection of DTC in peripheral blood. METHODS 32 cases of laryngeal or laryngopharyngeal carcinoma were investigated. DTC in peripheral blood was detected by nested reverse transcription polymerase chain reaction,using CK19mRNA as the marker. RESULTS In the RT-PCR study,15 of 32 cases (46.9 %) showed a positive result. Ten of the 25 cases (40 %) of laryngeal carcinoma were positive. Fix of the remaining 7 cases (71.4 %) of laryngopharyngeal carcinoma were positive. All controls were negative. Of the 20 cases without lymph node metastasis,6 were positive; of the 12 cases with lymph node metastasis,9 were positive. The positive rate of the group with lymph node metastasis was higher than that of the group without lymph node involvement(P

An explant outgrowth culture system for airway ciliated cells / 中国耳鼻咽喉头颈外科

OBJECTIVE To develop an explant outgrowth culture system for airway ciliated cells and explore the usefulness of the system.METHODS Mucosal specimens from rabbit tracheal epithelium were seeded into collagen-coated cover-slips. After 7 days,the specimens were characterized by Photo- contrast microscopy,transmission and Scanning electron microscope. They were also examined with hematoxylin-eosin staining and mucin-5AC immunohistochemistry. Ciliary beat frequency was measured with high-speed digital microscopy. RESULTS Many ciliated cells with good viability and goblet cells were found in the culture system. Immunohistochemistry analysis revealed that mucin- 5AC antibody labeled mainly in the goblet cells.The ciliated cells were differentiated well detected by electron microscope. At (30?1)℃,the basal ciliary beat frequency was (13.2?0.9) Hz. CONCLUSION The method may be useful in establishing a culture system for ciliated cells and the system will be suitable for the long-term studies of mucociliary transport system.

Preparation of rat tail collagen and a cell culture model of human nasal epithelial cells / 中国耳鼻咽喉头颈外科

OBJECTIVE To establish a cell culture of human nasal epithelial cells on glass cover slides coated with rat tail collagen and explore a method to research human mucociliary system. METHODS The rat tail collagen and collagen coated cover slides were prepared for explant culture of human nasal epithelial cells. RESULTS The collagen on cover slides must be thin and ? at. It was about 1mm thick. The cultured epithelial cells were monolayer and polygonal cells. There was tight junction between the cells. Ciliary beat frequency of different cilia was equal on the same cell,and it was also equal of on cells from uncinate process and inferior turbinate. CONCLUSION This successful cell culture model of human nasal epithelail cell could be helpful to research the physiology and function of human nasal mucociliary system.

Image-guided surgery in congenital bony aural atresia / 中国耳鼻咽喉头颈外科

OBJECTIVE Congenital aural atresia repair is difficult owing to unpredictable anatomy. Benefits may be gained from image-guided surgery(IGS) . its exact role and surgery indication were def ined. METHODS From 2001 to 2004,36 ears with bony type C(Schuknecht classification) congenital atresia were performed. In the IGS group(n=18) ,repair surgery was performed with IGS,while in the control group(Non-IGS,n=18) ,similar intervention was applied without IGS. IGS group:aged from 12-29 years,follow-up from 6 months to 1 year. Non-IGS group:aged from 10-27 years,follow-up from 6 months to 3 years. Intra-and post-operative clinical and audiometric findings were compared. RESULTS All of the patients had congenital bony aural atresia,ossicles malformation,tympanic cavity hypoplasia and facial nerve malformation. IGS revealed a malformed horizontal semicircular canal hidden in the bony atresia plate during the operation while computed tomography(CT) did not show preoperatively. IGS computed tomography images correlated well with intra-operative findings,gave the surgeon more securityand reduced operative time(2 hours and 24 minutes) by 25 minutes. The prepare time increased 20 minutes(15-30 minutes) ,but total time decreased 5 minutes in IGS group. The registration accuracy was 0.6-1.3 mm,average 0.84 mm,which was suitable for the otologic surgery. There were 1 case in IGS group and 3 cases in Non-IGS group happened local aural restenosis after operation. But there were no facial nerve paralysis and hearing injury happened in both groups,and all of the patients got the satisfactory hearing after the hearing reconstruction(the air-bone gap with an average of IGS is 31.8dB,Non-IGS is 30.5dB) . CONCLUSION In our estimation,IGS is valuable for type C congenital aural atresia repair. It serves as an educational tool and a guide both for the experienced and inexperienced surgeons in critical situations where anatomical landmarks are distorted and approach is limited. There is no statistically significant between two groups on hearing improvement after operation.

LOH and MSI of Fragile Histidine Triad Gene in laryngeal and hypopharyngeal carcinoma / 中国耳鼻咽喉头颈外科

0.05).⑥ The incidence of MSI at D3S1234,D3S4103,D3S1300 in recurring cases was 83.33 %,but the ratio of MSI inprimary cases was 30.77 %,the former was remarkablyhigher than the latter(P=0.004).CONCLUSION ①Microsatellite analysis showed that both LOH andMSI of FHIT gene existed in laryngeal carcinoma andhypopharyngeal carcinoma,the former was morecommon.② FHIT gene participates the developmenof laryngeal carcinoma and hypopharyngeal carcinoma and may be one of the candidate tumor suppressor genes.③ MSI of FHIT gene may be correlated with recurrence of laryngeal and hypopharyngeal carcinoma.

Function of MMP-2 and TIMP-2 in laryngeal carcinomas and the relationship among expression of MMP-2, TIMP-2 and microvessel density / 中国耳鼻咽喉头颈外科

OBJECTIVE To understand the function of MMP-2 and TIMP-2 and the relationship among expression of MMP-2, TIMP-2 and microvessel density in laryngeal carcinomas. METHODS The expression of MMP-2, TIMP-2 and CD34 in 37 laryngeal carcinomas patients were studied with immunohistochemical staining. The staining results were studied morphometrically with computer image analysis. RESULTS The mean values of MMP-2 expression and microvessel density (MVD) in squamous cell carcinoma group with lymph node metastasis were significantly higher than that without lymph node metastasis (P

Relationship between mutations of the hyper-variable region of mitochondrial DNA D-loop and clinical characteristics in laryngeal cancer / 中国耳鼻咽喉头颈外科

OBJECTIVE To investigate the features of somatic mutations of the hypervariable region (HV) of mitochondrial DNA (mtDNA) D-loop in laryngeal squamous cell carcinoma (LSCC) and the relationship with clinicopathological characteristics. METHODS Microdissected cancerous tissues, paracancerous tissues and peripheral vein blood were obtained from 40 patients with LSCC and total DNA was extracted. HVⅠ and HVⅡ fragments were amplified by PCR and purified. Mutations were determined by DNA sequencing. The mutations of HVⅠand HVⅡ and their relationship with clinicopathological characteristics were analyzed. RESULTS ①In 40 LSCC samples, 86 polymorphisms were found, 21 (52.5 %) tumors presented somatic mtDNA mutations with a total of 34 mutations. Among them, 28 (82.4 %) and 6 (17.6 %) were located in HVⅠ and HVⅡ respectively. ②Tumor D-loop mutation showed significant correlation with tumor differentiation(P=0.028) and p53 mutation (P=0.041), respectively. CONCLUSION Mitochondrial DNA D-loop in LSCC is an unstable region with high frequency of somatic mutation and high polymorphism and might play a potential role in the carcinogenesis of the larynx.