鼻息肉中碳酸酐酶IX表达与嗜酸细胞浸润相关性分析

目的研究碳酸酐酶IX（carbonic anhydrase IX,CAIX）在鼻息肉组织中表达及其与嗜酸细胞（eosinophils,Eos）浸润的相关性,探讨CAIX在鼻息肉发病中的作用。方法收集28例鼻息肉组织（鼻息肉组）和12例正常下鼻甲组织（对照组）,HE染色观察Eos浸润情况,免疫组化检测CAIX的表达。结果与对照组比较,鼻息肉组Eos浸润显著增加,CAIX免疫阳性细胞数及着色强度明显增高。图像分析显示,鼻息肉组CAIX的积分光密度（103/HP）为26.87±4.60,与对照组3.25±1.20相比,有显著性差异（P〈0.05）;Spearman等级相关分析显示,鼻息肉中CAIX表达与Eos浸润表达密切相关（r=0.632,P〈0.01）。结论鼻息肉组织中CAIX表达、Eos浸润均上调,二者有协同表达关系,说明鼻息肉组织中存在缺氧及缺氧适应现象,CAIX表达上调,从而使细胞适应缺氧微环境,保证缺氧区域细胞的持续生长和增殖,促进Eos浸润生长,导致一种慢性炎症反应。     

速尿对实验性膜迷路积水前庭碳酸酐酶活性的影响

为进一步探讨速尿对膜迷路脱水的机理，采用组织，细胞化学染色法，观察速尿对壶腹嵴毛细胞，前庭移行细胞，暗细胞及椭圆囊毛细胞碳酸酐酶的影响。结果显示，碳酸酐酶主要分布于半月面移行细胞侧膜和前庭暗细胞指状突胞膜；实验性膜迷路积水耳碳酸酐酶分布与正常耳无显著变化；速尿对前庭暗细胞，移行细胞的碳酸酐酶有明显抑制作用。     

Expression and distribution patterns of the stem cell marker,nestin, and the stem cell renewal factor,BMI-1, in normal human nasal mucosa and nasal polyps

Conclusions: These results suggest that nestin and BMI-1 are candidates for stem cell markers and renewal factors in human nasal mucosa, may contribute to tissue homeostasis and differentiation in the epithelium and submucosal glands of normal nasal mucosa, and may play a role in proliferation of nasal polyps. Objectives: The stem cell marker, nestin, and the stem cell renewal factor, BMI-1, have been identified in a variety of inflammatory and normal tissues, implicating these markers in tissue regeneration. Materials and methods: We investigated the expression and distribution of nestin and BMI-1 in normal nasal mucosa and nasal polyps, using RT-PCR, immunohistochemistry, and Western blotting. Results: Nestin and BMI-1 were localized to the epithelium and submucosal glands of normal nasal mucosa and nasal polyps. The expression of nestin was confined to the plasma membrane and cytoplasm, whereas BMI-1 showed a nuclear staining pattern. In normal nasal mucosa and nasal polyps, nestin and BMI-1 expression was strongest in the basal portion of the epithelial layer, and decreased toward the upper portion. In the submucosal glands, weak to strong expression was commonly detected in the glandular acini. There was no significant difference in the level of expression of nestin and BMI-1 between normal nasal mucosa and nasal polyps.     

Nods样模式识别受体在鼻息肉组织中的表达及意义

目的：探讨Nods（核苷酸结合寡聚化结构域,Nod Like receptors）样模式识别受体（PRR）在鼻息肉组织中的表达及作用。方法：分别采用实时定量RT—PCR、Western—Blot及免疫组织化学检测Nodl、Nod2、Nalp3mRNA及蛋白在鼻息肉组织中的表达（以正常下鼻甲黏膜作对照）。结果：实时定量RT—PCR显示,鼻息肉组织Nodl、Nod2、Nalp3mRNA表达与正常对照下鼻甲黏膜比较无显著差异。Western-Blot结果提示：鼻息肉组织Nodl表达高于对照组,两组Nod2、Nalp3无明显差异。免疫组织化学表明,三种模式受体在两组中均有表达,主要表达于上皮细胞、腺体上皮、炎性细胞（如嗜酸粒细胞）,但Nodl及Nalp3在鼻息肉中表达强于对照组（P〈0．05）,Nod2蛋白表达无显著差异。结论：在鼻息肉组织中,Nodl、Nod2及Nalp3均有表达,其中Nodl、Nalp3的表达高于对照组,说明Nodl、Nalp3有可能参与鼻息肉的发病。     

Effects of topical Amphotericin B on expression of cytokines in nasal polyps

Objective—Although chronic rhinosinusitis (CRS) is one of the most frequently reported chronic diseases its etiology is not well understood. Recently, fungi have been proposed to influence the chronicity of rhinosinusitis. If fungi do play an important role then topical antifungal treatment may improve the inflammatory process of CRS. Therefore, in this study we measured inflammatory cytokine levels in nasal polyps after intranasal antifungal irrigation.

Material and Methods—Nasal polyps were collected before and 4 weeks after treatment with 100 mg/l topical amphotericin B (n=16), 50 mg/l topical amphotericin B (n=14) or normal saline (n=11). The cytokine—IL-5, IL-8, interferon-γ, RANTES—protein content of polyp homogenates were determined by means of ELISA.

Results—Nasal polyps were found to contain large amounts of cytokines (IL-5, IL-8 and RANTES) compared with normal inferior turbinates. After 4 weeks of treatment with topical agents, IL-5 levels tended to decrease in comparison with those of the other cytokines, but this difference was not statistically significant.

Conclusions—Topical amphotericin B treatment and nasal saline irrigation both influence the expression of nasal polyp cytokines. Topical nasal irrigation may influence the inflammatory process of CRS.     

冲击波对豚鼠耳蜗毛细胞碳酸酐酶的影响

用组织化学、细胞化学方法结合图像分析和听觉电生理技术,研究了冲击波对豚鼠耳蜗毛细胞碳酸酐酶（CA）的影响及其与听阈阈移的关系。结果表明冲击波暴露后6、12、24、48、72小时组,毛细胞中CA阳性产物的灰度值较正常组明显降低（P＜0．01）,其灰度值的变化与听觉脑干反应（ABR）阈移呈负相关（r=-0．9369,P＜0．01）。提示毛细胞中CA活性减低是冲击波致听力损失的因素之一。文中讨论了CA在耳蜗毛细胞的作用及其与听阈的关系。     

Ultrastructural localization of carbonic anhydrase in the vestibular end organs of the guinea pig

Summary Carbonic anhydrase activity was demonstrated cytochemically on an ultrastructural level in the vestibular end organs of the guinea pig. Reaction product was found in the dark cells, transitional cells, cells of the planum semilunatum and supporting cells. In the dark cells, reaction product was observed in the cytoplasm as well as in the basal infoldings. Reaction product was also observed in the basal infoldings of the transitional cells and the cells of the planum semilunatum. The globular structures inside the supporting cells, transitional cells and the cells of the planum semilunatum were also surrounded by the reaction product. These findings suggest that carbonic anhydrase may have different functions, such as water and ion transport, respiration, nutrition and calcium carbonate deposition in the vestibular end organs.     

Role of free radicals and antioxidants in nasal polyps

OBJECTIVES/HYPOTHESIS: The aim of this study is to determine the role of free radicals and antioxidants in nasal polyps. STUDY DESIGN: Prospective, randomized, controlled study. METHODS: Thirty-one patients with nasal polyposis and a control group consisting of 19 patients with septal deviation and lower turbinate hypertrophy were included in the study. Levels of the antioxidants retinol, beta-carotene, alpha-tocopherol, and ascorbic acid were measured from the sera of the patients with nasal polyposis and the control group. Plasma levels of superoxide dismutase activity (SOD), glutathione peroxidase (GSHPX) activity, and reduced glutathione (GSH) were also obtained. As a peroxidation product, the levels of the malondialdehyde-thiobarbituric acid (MDA) combination were measured from the plasma of patient and control groups. Measurements of MDA, GSH, and alpha-tocopherol levels were also taken from the polyp tissue and turbinate mucosa of the control group. RESULTS: The blood levels of antioxidants and MDA as an oxidant were significantly different in the patient group compared with the control group (P <.01). The tissue levels of antioxidants and MDA were significantly different in the patients with polyposis compared with the control group (P <.01). The blood and tissue anti-oxidant levels were found to be decreased, and MDA levels as an oxidant increased significantly in the patient group with polyposis when compared with the control group, and there was a negative correlation between oxidative stress and antioxidants. CONCLUSION: This study demonstrates that oxidative stress and tissue and blood antioxidants in the patients with polyposis were significantly different compared with the control group. The blood and tissue antioxidant levels decreased, and MDA levels, as an oxidant, increased significantly in the patient group with polyposis when compared with the control group. The current study demonstrates that there is strong evidence related to oxidative stress in the pathogenesis of nasal polyposis, and antioxidants can have a preventive role in free-radical-mediated tissue damage in nasal polyposis.     

Prevalence of Helicobacter pylori in patients with nasal polyps: a preliminary report

OBJECTIVES: The aim of the study is to determine the presence of H. pylori in nasal polyps by both immunohistochemical staining with H. pylori antibody of biopsy specimens and enzyme-linked immunoadsorbent assay (ELISA) of sera. STUDY DESIGN: A prospective, controlled, clinical trial. METHODS: We enrolled 30 patients with nasal polyps and 20 controls with middle concha bullosa undergoing endoscopic sinus surgery (ESS). Blood samples of both the study and control groups were evaluated for anti-H. pylori specific immunoglobulin (Ig)G antibodies by ELISA. In addition, biopsy specimens of the removed polyps and the mucosal part of middle conchas were examined by the immunohistochemical analysis with H. pylori antibody. RESULTS: In the blood samples, specific IgG antibodies to H. pylori were found in 26 (86.7%) of 30 polyp patients and 17 (85%) of 20 controls. In 6 (20%) of the 30 patients, H. pylori was identified in the nasal polyp tissue, but it was not detected in the mucosal part of the middle concha specimens. No significant statistical difference was observed for H. pylori antibodies by ELISA among the patients with nasal polyps and the control group (Fisher's exact test, P = .59). However, there was a statistical difference between the polyp biopsy specimens and the control biopsy specimens by immunohistochemical staining (Fisher's exact test, P = .037). CONCLUSIONS: This study indicates that H. pylori was found in increased prevalence in the nasal polyps. However, further controlled epidemiologic studies would be necessary to confirm our results and clarify the potential underlying pathogenetic mechanisms.     

嗜酸性粒细胞与慢性鼻-鼻窦炎伴鼻息肉的相关性研究进展

慢性鼻窦炎（chronic rhinosinusitis,CRS）是鼻腔黏膜和鼻窦黏膜的慢性炎性疾病,根据有无鼻息肉可分为慢性鼻-鼻窦炎伴鼻息肉（chronic rhinosinusitis with nasal polyps,CRSwNP）和慢性鼻-鼻窦炎（chronic rhinosinusitis without nasal polyps,CRSsNP）。鼻息肉组织常见有嗜酸性粒细胞(eosinophils,EOS) 浸润增多,根据嗜酸性粒细胞的浸润程度可将CRSwNP分为两个亚型：嗜酸粒细胞型慢性鼻窦炎伴鼻息肉(eosinophilic chronic rhinosinusitis with nasal polyps, ECRSwNP)和非嗜酸粒细胞型慢性鼻窦炎伴鼻息肉 (nonesinophilc chronic rhinosinusitis with nasal polyps,nonECRSwNP);将慢性鼻窦炎分为：嗜酸粒细胞型慢性鼻窦炎（eosinophilic chronic rhinosinusitis, ECRS)和非嗜酸粒细胞型慢性鼻窦炎(non eosinophilic,NECRS)。嗜酸性粒细胞在鼻息肉形成、发展机制中的作用多年来一直备受学者们关注。随着研究的不断深入,嗜酸性粒细胞在鼻窦炎鼻息肉炎症趋化、息肉形成、分型和预后中的作用得到越来越多的重视。     

不伴鼻息肉的慢性鼻-鼻窦炎鼻黏膜组织重塑研究

目的：观察慢性鼻-鼻窦炎不伴鼻息肉（cRssNP）黏膜组织重塑的特点。方法：收集30例CRSsNP患者和10例对照组的钩突黏膜组织,分别进行苏木精一伊红染色、Masson三色胶原染色、阿辛蓝-过典酸-希夫染色及苦味酸-天狼星红染色,观察黏膜上皮损伤、杯状细胞化生、基膜增厚、细胞外基质胶原沉积、腺体增生及胶原组成等情况。结果：①CRSsNP组上皮细胞损伤1级、2级和3级均比对照组明显增多（P〈0．01或P〈0．05）,而损伤0级比对照组明显减少（P〈0．01）。②黏膜下固有层黏液腺增生明显,分泌旺盛,上皮层杯状细胞计数较对照组明显增多（P〈0．01）。③黏膜细胞外基质胶原沉积较对照组明显增多,基膜明显增厚（P〈0．01）。④胶原沉积以Ⅰ型胶原为主,Ⅲ型、Ⅳ型胶原较少。结论：CRSsNP存在明显的鼻黏膜组织重塑,表现为上皮细胞损伤,细胞外基质胶原沉积、基膜增厚、杯状细胞和黏液腺增生。     

变应性因素及变应性鼻炎与慢性鼻-鼻窦炎鼻息肉的相关性探讨

目的：探讨变应性因素及变应性鼻炎（AR）与慢性鼻-鼻窦炎鼻息肉的临床相关性。方法：将200例研究对象分为A、B组,A组诊断为AR（110例）,B组诊断为慢性鼻-鼻窦炎鼻息肉（90例）,通过欧蒙印迹法定量检测血清sIgE浓度并观察慢性鼻-鼻窦炎鼻息肉手术治疗后的复发率、AR并发慢性鼻-鼻窦炎鼻息肉的发生率。结果：A组sIgE总阳性率为89.09%,B组sIgE总阳性率为74.44%。B组中sIgE阳性者术后复发率为58.21%,sIgE阴性者术后复发率为8.70%。A组41例（37.27%）并发慢性鼻-鼻窦炎鼻息肉,血清sIgE阳性率为97.56%;69例（62.73%）未并发慢性鼻-鼻窦炎鼻息肉,血清sIgE阳性率为79.71%;组内slgE阳性率比较差异有统计学意义（χ2=6.96,P〈0.01）。结论：变应性因素及AR与慢性鼻-鼻窦炎鼻息肉的发生有一定的相关性,提示避免接触变应原、合理治疗AR能够有效控制慢性鼻-鼻窦炎鼻息肉的复发率。     

克拉霉素对离体鼻息肉组织中环氧合酶-2及核因子κB表达的影响

目的：检测用不同浓度梯度的克拉霉素干预后,离体鼻息肉组织中环氧合酶-2（COX-2）及核因子κB（NF-κB）的表达状况,探讨其在鼻黏膜炎症机制中的作用。方法：将鼻息肉组织块分别与克拉霉素（0、10^-6、10^-5及10^-4mol／L）于体外共同培养1d,应用Western blot和荧光实时定量PCR技术检测COX-2和NF-κB亚基的表达水平,观察克拉霉素干预后COX-2和NF-κB表达的变化规律。结果：对照组（0mol／L克拉霉素组）鼻息肉组织中COX-2、NF-κBp50和NF-κcBp65的蛋白质和核酸表达水平最高;随着克拉霉素干预剂量的增加,COX-2、NF-κBp50和NF—κBp65的表达水平呈剂量依赖性下降。相关分析表明,同一组鼻息肉组织中,COX-2核酸表达量分别与NF-κBp50、NF-κBp65呈直线正相关。结论：COX-2异常表达参与了鼻-鼻窦黏膜慢性炎症反应,克拉霉素可能通过阻断NF-κB通路来下调COX-2的合成,发挥其抗炎作用。     

D2-40 immunohistochemical assessment of lymphangiogenesis in normal and edematous sinus mucosa and nasal polyp

OBJECTIVE: The lymphatic system plays an important role in the maintenance of tissue fluid homeostasis, which facilitates interstitial protein transport. Until recently, the lymphatic system of the sinonasal mucosa has been relatively poorly studied. The authors aimed to investigate the distributional and quantitative changes of the lymphatic vessels in inflammatory sinus mucosa and nasal polyps in comparison with healthy sinus mucosa using D2-40 antibody. METHODS: Immunohistochemistry and Western blotting with D2-40 antibody were applied to normal and edematous ethmoid sinus mucosa and nasal polyps. The number, size, and length densities of lymphatic vessels were evaluated using tissue sections and whole mount preparations. RESULTS: Most lymphatic vessels in normal and edematous ethmoid sinus mucosa were distributed in the subepithelial layer. Some of these vessels were dilated, whereas others were compressed and had a slit-like lumen. No D2-40 positive vessels were found in samples of nasal polyps. Lymphatic vessels showed no statistically significant difference in their number, size, or length density between normal and edematous ethmoid sinus mucosa. Western blot also showed no differences in their expression levels. CONCLUSION: These findings indicate that lymphangiogenesis does not occur in edematous ethmoid sinus mucosa, which may not reuptake interstitial fluid efficiently in inflammatory conditions, resulting in the formation of mucosal edema in chronic inflammation.     

Comparison of subjective and objective assessment of glucocorticoid response in nasal polyps: a preliminary study

Background: Glucocorticoids (GC) therapeutic response in patients with chronic rhinosinusitis with nasal polyps (CRSwNP) varies markedly.

Aims/Objectives: To compare the utility between subjective and objective assessment of GC sensitivity in reflecting the impact of GC on systemic and local eosinophilia in CRSwNP patients.

Material and methods: Twenty-six patients with CRSwNP were enrolled. All patients were given 30?mg of prednisone once daily for 7 days and subsequently classified into subjectively GC-sensitive and -insensitive subgroup or objectively GC-sensitive and -insensitive subgroup. The numbers of eosinophils and neutrophils in blood and polyp tissues were compared between GC-sensitive and GC-insensitive subgroup.

Results: 17/26 (65.4%) patients were subjectively and 8/26 (30.8%) patients objectively sensitive to GC treatment. The absolute number and percentage of eosinophils in blood were decreased both in GC-sensitive and -insensitive subjects after GC treatment. In addition, a significant reduction in tissue eosinophil percentage was only observed in objectively GC-sensitive subjects after GC treatment. Furthermore, the change of tissue eosinophil percentage in objectively GC-sensitive subjects was significantly higher than that in objectively GC-insensitive subjects.

Conclusions and significance: Objective assessment may better reflect oral GC response in tissue eosinophilic inflammation than subjective assessment in patients with CRSwNP.     

The nasal response to axillary pressure

The changes in ipsilateral sympathetic tone in response to axillary pressure are well known. However, the response of the nasal mucosa is less well documented. We have attempted to demonstrate this response in normal individuals. Ten individuals with no history of nasal disease or allergy were studied. All subjects were exposed to sustained pressure, using a crutch, for a period of 5 min. Nasal cross-sectional area was measured on both sides of the nose using an acoustic rhinometer along with pulse and blood pressure. The individuals were then rested for at least 30 min and the test repeated with pressure applied to the opposite side. Statistical analysis was performed by non-parametric methods. There was a significant fall in nasal cross-sectional area on the side of pressure (median change = 0.09 cm², P < 0.01) while cross-sectional area in the contralateral nasal passage increased (median change = 0.35 cm², P = 0.01). There were no significant differences between these results and those obtained by axillary pressure on the opposite side. Pulse and diastolic blood pressure rose with axillary pressure while systolic blood pressure remained unaltered. There was no difference in the laterality in the blood pressure response. The results indicate that axillary pressure produces nasal congestion and both afferent and efferent arms of this reflex are side-specific.     

水通道蛋白-4在人鼻息肉组织中的表达及意义

目的 研究水通道蛋白-4(aquaporin4, AQP-4)在鼻息肉组织中的分布并探讨其在鼻息肉形成过程中的机制和作用。方法 采用免疫组织化学方法检测60例新鲜鼻息肉标本和30例肥大下鼻甲黏膜中AQP-4的表达及分布水平,应用两独立样本t检验对检测结果进行对比分析。结果 鼻息肉和下鼻甲黏膜的上皮细胞、固有层腺体细胞、血管及血窦内皮中均可检测到AQP-4分布;鼻息肉上皮细胞、血管及血窦内皮中阳性细胞数均高于下鼻甲黏膜,且差异有统计学意义。结论 AQP-4在鼻息肉上皮细胞和血管及血窦内皮细胞中表达水平明显增高,提示AQP-4在鼻息肉组织中的异常分布可能是鼻黏膜水肿的形成机制之一。