Effect of RNA interference therapy on the mice eosinophils CCR3 gene and granule protein in the murine model of allergic rhinitis

Objective:To observe the clinical manifestations of allergic rhinitis mice and the expression changes of the eosinophils CCR3 and the granule protein rnRNA in the bone marrow,peripheral blood and nasal lavage fluid.Mctliods:Twenty-four BALB/c mice were randomly divided into the control group.PBS therapy group.siKNA therapy group and the CCR3 siRNA therapy group(n=6).Allergic rhinitis model were sensitized and stimulated by ovalbunfin,and CCR3 siKNA therapy group were administered with CCH3 transnasally before stimulated.The levels of the eosinophils CCR3.MBP.ECP and EPO in bone marrow,peripheral blood and nasal lavage fluid were detected by RT-PCR.Results:Compared to the control group and CCR3 siR.NA therapy group,the nasal mucosa of the PBS therapy group and siRNA therapy group developed epithalaxy.goblet cells hyperplasia,squamous epithelium metaplasia,epithelium necrosis,lamina propria and submucosa gland hyperplasia,vasodilatation,tissue edema,and the characterized eosinophil infiltration.RT-PCR indicated that the CCR3 rnRNA,MBP.ECP and EPC)expression in bone marrow,peripheral blood and nasal lavage fluid of the CCR3 siKNA therapy group was lower than the PBS therapy group and siR.NA therapy group(P<0.05).Conclusions:The RNA interference therapy to CCR3 by local administration pernasal can suppress the process of the development,migration and invasion of the allergic rhinitis eosinophil,thus can reduce the effect of eosinophils and then reduce the inflammation effect of the allergic rhinitis.It may be a new treatment for respiratory tract allergic inflammation.

Effect of Centipeda Herba Volatile Oil on Allergic Rhinitis by Nasal Sniffing

[Objectives] To explore the efficacy of different concentrations of Centipeda Herba volatile oil in the treatment of allergic rhinitis (AR) by nasal sniffing and the optimal drug concentration.[Methods] Forty-eight mice were randomly divided into control group, blank group, alcohol treatment group, low concentration group, medium concentration group and high concentration group, with 8 mice in each group. Except for the blank group, each mouse in the other groups was intraperitoneally injected with 1 mL of a mixture of 50 μgOVA+5 mg [Al(OH) 3] +1 mL of normal saline for 14 d. The allergic rhinitis mouse model was successfully established by intranasal instillation of 5% OVA solution on both sides (20 μL per side, once a day) from the 15 th day after stimulation for 7 d. The blank group was treated with the same amount of saline as above. The volatile oil of Centipeda Herba was obtained by steam distillation and petroleum ether extraction, and then was made into 1.25%, 2.5% and 5% volatile oil of Centipeda Herba with 75% alcohol. The control group was stimulated once every other day with reagent 2 after 7 d of stimulation (maintenance) until the end of treatment. The blank group was treated with 5 mL saline by nasal sniffing for 30 d, twice in the morning and evening, 30 min each time. The alcohol treatment group was treated with 5 mL of 75% alcohol, and the low concentration group, the medium concentration group and the high concentration group were treated with 5 mL of 1.25%, 2.5% and 5% Centipeda Herba volatile oil, respectively. The treatment time was the same as that of the blank group, and the treatment process was carried out in their respective closed contamination boxes. Before and after the treatment, the frequency of sneezing, the frequency of scratching nose, the amount of nasal discharge, activity and other general characteristics of the mice were observed, and the allergic behavior score was carried out. Besides, the IgE content in the serum of the mice was determined, and the eosinophils in the nasal discharge were counted.[Results] The scores of mice before and after treatment showed that there was no significant difference in the alcohol treatment group before and after treatment ( P >0.05), and there was significant difference in the low, medium and high concentration groups before and after treatment ( P <0.05), except that there was no significant difference between the control group and the alcohol treatment group ( P >0.05), and there was significant difference among the other groups ( P <0.05). The levels of IgE and the number of eosinophils in peripheral serum of mice in the control group, alcohol treatment group, low concentration group, medium concentration group and high concentration group were higher than those in the blank group ( P <0.05), and there was no significant difference between the two groups ( P >0.05).[Conclusions] Volatile oil of Centipeda Herba can be used to treat allergic rhinitis by nasal sniffing, and 5% volatile oil of Centipeda Herba has the best effect. During the treatment, sneezing and runny nose in mice were reduced. The results showed that nasal sniffing was less irritating to the nasal cavity and not easy to produce discomfort, the utilization rate of drugs was higher than that of traditional therapy, and the volatile oil could be preserved longer than that of traditional Chinese medicine.

OBSERVATION OF REMOTE AND RECENT THERAPEUTIC EFFECTS OF ACUPOINT-THREAD EMBEDDING, WESTERN MEDICINE OR ALLIED THERAPY ON PERENNIAL ALLERGIC RHINITIS

Objective: The comparison of the remote and recent therapeutic effects on perennial allergic rhinitis was carried on among single application of acupoint-thread embedding, western medicine (certirizine+flixonase) and allied therapy. Method: Ninety cases were randomized into thread-embedding group, western medicine group and embedding+western medicine group (allied-therapy group), 30 cases in each one. In thread embedding group, Yingxiang (迎香 LI 20) and Feishu (肺俞 BL 13) were selected for thread embedding, once a week, totally 4 treatments were required. In western medicine group, certirizine tablet 10 mg in combination with flixonase sprayer was applied once per day, continuously for 30 days in treatment. In allied-therapy group, both of the above methods were adopted continuously for 30 days in treatment. The therapeutic effects were in statistics at the end of treatment and 1 year later. Results: At the end of treatment, in thread-embedding group, 21 cases were remarkably effective, 3 cases effective and 6 cases no effect. In western medicine group, 20 cases were remarkably effective, 3 cases effective and 7 cases no effect. In allied-therapy group, 23 cases were remarkably effective, 2 cases effective and 5 cases no effect. It was shown in statistics that there was no significant difference in therapeutic effects among three groups ( P>0.05 ). But one year after the end of treatment, it was indicated that in thread-embedding group, 3 cases were remarkably effective, 10 cases effective and 17 cases no effect; in western medicine group, 2 cases were remarkably effective, 9 cases effective and 19 cases no effect; in allied-therapy group, 7 cases were remarkably effective, 11 cases effective and 12 cases no effect. The significant difference in the therapeutic effects had not been presented in the comparison between thread-embedding group and western medicine group (P>0.05), but that had been presented between allied-therapy group with either of the groups with single therapy applied (P<0.05). Conclusion: Quite good recent therapeutic effect of either acupoint-thread embedding or western medicine (certirizine+flixonase) was achieved on perennial allergic rhinitis. The remote therapeutic effect of allied therapy was more advantageous than that of single application of either thread embedding or western medicine.

Observation of Ultrastructure in Nasal Mucosa on Allergic Perennial Rhinitis Cases before and after the Operations

Objective: The partial inferior turbinectomy and septoplasty was applied to treat the allergic perennial rhinitis (APR), and to observe the ultrastructure changes of the nasal mucosa before and after the operations. Methods: For 36 cases of research objects diagnosed with APR, the partial inferior turbinectomy and septoplasty was administered. For 6 APR cases among them, the pre- and postoperative observation of anterior nasal mucosa of the inferior turbinate on the same side under the electron microscope was conducted for one year respectively. In addition, their pathological alterations of tissues were also conducted. Results: In the pre-operative observation under the electron microscope, it was found that the nasal mucosae epithelium cells were nude without cilia. The lamina propria had edema, and mesenchyme presented the infiltration of substantial eosinophilic granulocytes, basophilic granulocytes, plasmacytes and mast cells. Additionally, abundant degranulation and vacuolation of cytoplasts were observed. The plentiful glands, duct ectasia, edema and structural changes were also found. Some gland cells had degenerated. After the operation, it was found that the epithelium nudity still existed and the deficiency of cilia was not improved. Only a very small amount of microvilli existed. The edema of lamina propria was improved and eosinophilic granulocytes were rarely observed in mesenchyme. However, the infiltration of basophilic granulocytes, plasmocytes and mast cells was still observed. The particle structure was generally stable and the central crystal was clear without degranulation. Meanwhile, the amount of glands was reduced and the tissue structure tended to be recovered. Overall, the nasal mucosa showed changes as chronic inflammation. Conclusions: For the treatment of APR with the methods presented by our research institution, in one year before and after the operation, ultrastructural changes of inferior turbinate mucosa tissues were observed from the preoperatively pathological changes of typical APR to the chronic inflammation with the primary infiltration of neutrophilic granulocyte and mast cells.

Study on Cytokines IL-2,IL-6,IL-10 in Patients of Chronic Allergic Rhinitis Treated with Acupuncture

OBJECTIVES: To observe the plasmatic concentration of IL-6, IL-10 and IL-2 in the patient of chronic allergic rhinitis before and after acupuncture therapy. METHODS: Cytokine levels were determined before and after treatment in 30 healthy volunteers (Group A) and 90 patients of chronic allergic rhinitis (Group B) with an increased plasma IL-10 level. Group B was then divided into 3 subgroups: 30 patients treated with real acupuncture (Group B1); 30 patients treated with sham acupuncture (Group B2); 30 non-treated patients (Group B3). RESULTS: The allergic subjects of group B1, compared with controls, showed a significant reduction of IL-10 after a specific treatment with acupuncture (P

Clinical Research into Ke Min Yin for Treatment of Persistent Allergic Rhinitis in Patients with Qi Deficiency and Blood Stasis

Persistent allergic rhinitis (PAR) is one of the common diseases at ear-nose-throat department. From August 1998 to May 2001, we randomly selected 60 cases of PAR with qi deficiency and blood stasis for a clinical observation. 30 patients in the treatment group were treated with Ke Min Yin (克敏饮), a herbal medicine, to supplement qi, expel wind, nourish blood and promote blood circulation; and the other 30 cases in the control group were treated with cetrine. Through systematic observation and statistical processing, the clinical research is summarized and reported in the following.

A CLINICAL OBSERVATION ON THERAPEUTIC EFFECTS OF ACUPUNCTURE FOR ALLERGIC RHINITIS

The author applied acupuncture to treat52 cases of allergic rhinitis with certaintherapeutic effectiveness confirmed by afollowing-up for one year and more.The resultis reported as follows.

Effects of lysedEnterococcus faecalis FK-23 on experimental allergic rhinitis in a murine model

In the current study, we sought to investigate whether lysed Enterococcus faecalis FK-23 （LFK）, a heat-killed probiotic preparation, attenuated eosinophil influx into the upper airway and had immunomodulatory activity in a murine allergic rhinitis model. Eighteen BALB/c mice were divided into three groups; the ovalbumin （OVA）-sen- sitized/challenged group, which received saline orally for 6 weeks （OVA group）, the OVA-sensitized/challenged group, which received LFK orally for 6 weeks （LFK-fed group）, and the non-sensitized group, which received saline for 6 weeks （saline control group）. Nasal rubbing and sneezing were monitored during the study. After the final challenge, interleukin （IL）-4, interferon （IFN）-y, and OVA-specific IgE levels in the sera and splenocyte culture supernatants were determined, eosinophilic infiltrate into the upper airway was quantified, and splenic CD4~CD25＋ regulatory T cells （Tregs） were examined by flow cytometry. We found that nasal rubbing was sig- nificantly reduced in LFK-fed mice compared to the OVA group on d 27 and 35, and sneezing was significantly inhibited by LFK administration for 35 d. LFK-fed mice had significantly less eosinophil influx into the nasal mucosa than the OVA group. There were no significant differences between the LFK-fed group and OVA group in the serum and splenocyte culture supernatant levels of IL-4, IFN-y, and OVA-specific IgE. Interestingly, the LFK-fed mice had a significantly greater percentage of splenic CD4＋CD25＋ Tregs than OVA group. Our results indicate that oral administration of LFK may alleviate nasal symptoms, reduce nasal eosinophilia, and increase the percentage of CD4＋CD25＋ Tregs in experimental allergic rhinitis.

Correlation of serum YKL-40 content with eosinophil function as well as Th1/Th2 and Treg/Th17 immunity in patients with allergic rhinitis

Objective: To discuss the correlation of serum YKL-40 content with eosinophil function as well as Th1/Th2 and Treg/Th17 immunity in patients with allergic rhinitis. Methods: A total of 310 patients with allergic rhinitis who were treated in the hospital between January 2015 and May 2017 were selected as allergic rhinitis group and 100 healthy volunteers who received physical examination in the hospital during the same period were selected as normal control group. The differences in serum YKL-40 content, eosinophil function as well as Th1/Th2 and Treg/Th17 cytokine contents were compared between the two groups. Pearson test was used to assess the correlation between serum YKL-40 content and disease severity in patients with allergic rhinitis. Results: Serum YKL-40 content of allergic rhinitis group was higher than that of normal control group;serum eosinophil function IgE and Eotaxin-2 contents of allergic rhinitis group were higher than those of normal control group;serum Th1 cytokines IFN-γand IL-12 contents were lower than those of normal control group while Th2 cytokines IL-4 and IL-13 contents were higher than those of normal control group;serum Treg cytokine TGF-β content was lower than that of normal control group while Th17 cytokine IL-17 content was higher than that of normal control group. Pearson test showed that serum YKL-40 content in patients with allergic rhinitis was directly correlated with the eosinophil function as well as Th1/Th2 and Treg/Th17 immunity. Conclusion: Serum YKL-40 content is abnormally high in patients with allergic rhinitis, and the specific expression is positively correlated with the disease severity.

Treatment of 48 Cases of Perennial Allergic Rhinitis with Thunder-fire Moxibustion

Perennial allergic rhinitis is an allergic reaction of the nasal mucosa. It is characterized by paroxysmal and repeated runny nose, sneeze, stuffy nose, rhinocnesmus and nasal mucosa edema. The author used ＂Doctor ZHAO＇s thunder-fire moxibustion＂ in treating 48 cases of this disease. Now it is reported as follows.

Effects of Acupuncture-moxibustion on Monocyte Th1/Th2 Cytokine in Peripheral Blood of Patients with Perennial Allergic Rhinitis

Objective： To observe the effects of acupuncture-moxibustion on symptoms, signs, and Th1/Th2 cytokine, and regulation on Th1/Th2 in patients with perennial allergic rhinitis（PAR） to provide experimental basis for acupuncture treating allergic rhinitis and investigate the mechanism. Methods： Thirty patients with allergic rhinitis were treated with acupuncture-moxibustion, and 30 healthy cases were the control. Clinical symptoms, signs, and nasal mucosa were observed, and contents of IL-4, IFN-γ, and granulocyte-macrophage colony stimulating factor （GM-CSF） in the supernatant of peripheral blood monocyte cultured with PHA by ELISA, before treatment, after one course of treatment, and after two courses of treatment. Results： Before the treatment, the contents of IL-4, and GM-CSF were higher （P〈0.01） and IFN-γ was lower （P〈0.01） in the PAR cases than in the control cases. In the PAR cases, the content of GM-CSF was positive correlation with that of IL-4 （P〈0.01）, and negative correlation with IFN-7 （P〈0.01）. After the treatment, contents of IL-4 and GM-CSF were lowed （P〈0.01）, content of IFN-γ was increased （P〈0.05）, and all IL-4, GM-CSF and IFN-γ had no difference compared with those in the control cases. Conclusion： Acupuncture- moxibustion could improve the symptoms and signs in PAR patients, enhance the immune function. It may be achieved by regulating the expression of Th1 and Th2, correcting the imbalance of Th1/Th2.

Vesiculation Therapy of Cantharides Extract forPerennial Allergic Rhinitisand Its Effect on Serum Tolal IgE

The therapy of 10% Cantharides extract in treating 50 cases of perennial allergic rhinitis(PAR) was studied. The extract was plastered and blistered on Dazhui and Neiguan points. It was observedby nasal mucosa provocative test, cells in nasal secretion test and serum total IgE test. The results showedthat ite effective rate was 88% . After treatment the allergic nasal mucosa provocative test of the treatedgroup was alleviated obviously ( P < 0 . 01 ) and the number ot eosinopnil affd basophil in nasal secretion de-creased significantly ( P < 0 . 01 , P < 0 . 05 ) . The serum total lgE also reduced significantly I P < 0 . 01 ) .

The Decrease of Lung Eosinophil Following Zinc Supplementation in Mouse Models of Respiratory Allergic Inflammation

Background:United airway disease,a comorbid of allergic rhinitis and asthma,leads to the accumulation of toxic eosinophil in the lung parenchyma as a result of allergic reaction.Ovalbumin(OVA)sets off a condition of eosinophilia in the respiratory tract.Zinc supplementations are proposed to decrease the amount of eosinophil in the lung parenchyma due to its cytoprotective,anti-inflammation and antioxidant activity.Objectives:Proving the effects of the zinc supplementation on the number of eosinophil in the lung histopathology preparations of the BALB/c mice that were sensitized with OVA.Method:A post-test only control group design was used,which consisted of eighteen female BALB/c mice divided into three groups:the negative control group(K1),positive control group with OVA sensitization(K2),and group of mice with OVA sensitization and 5 mg/kg body weight zinc supplementation(P).The treatments were given for 30 days and then eosinophils in the lung histopathology preparations of the BALB/c mice were counted.Results:The mean eosinophil count in the negative control group(K1)was 0.48,the positive control group(K2)was 2.12,and the group of mice with OVA sensitization and 5 mg/kg body weight zinc supplementation(P)was 0.76.The number of eosinophil in the lungs in the K2 group was significantly higher than the K1 group(p=0.000)and the P group was significantly lower than K2 group in ANOVA Test followed by Post Hoc Test(p=0.001).Conclusion:The zinc supplementation reduces the number of eosinophil in the lungs tissue of the BALB/c mice that were sensitized with OVA.

呼出气一氧化氮、肺泡一氧化氮和嗜酸性粒细胞对3~6岁儿童呼吸系统疾病的鉴别诊断价值

目的探讨呼出气一氧化氮(FeNO)、肺泡一氧化氮(CaNO)和嗜酸性粒细胞(EOS)在甘肃省兰州市3~6岁儿童呼吸系统疾病鉴别诊断中的应用价值。方法选取确诊哮喘或过敏性鼻炎或下呼吸道感染的360例3~6岁儿童作为研究对象,采用斯皮尔曼秩相关系数评估FeNO、CaNO、EOS的相关性,通过随机森林模型、受试者工作特征(ROC)曲线、多因素逻辑回归分析评估FeNO、CaNO和EOS对3种疾病的鉴别诊断价值。结果哮喘患儿的FeNO、CaNO中位数高于其他疾病患儿,过敏性鼻炎患儿的EOS中位数最低,下呼吸道感染患儿的FeNO、CaNO中位数最低。相关性分析结果显示,FeNO与CaNO呈正相关(r=0.59,P<0.05),FeNO与EOS呈负相关(r=-0.61,P<0.05),CaNO与EOS呈负相关(r=-0.63,P<0.05)。随机森林模型显示,FeNO在疾病分类中的重要性最高。ROC曲线分析结果显示,3种疾病中,FeNO、CaNO、EOS对下呼吸道感染的诊断效能均最高(曲线下面积分别为0.86、0.91、1.00)。多因素逻辑回归模型诊断哮喘的曲线下面积为0.96,灵敏度为0.902,特异度为0.881。结论FeNO、CaNO和EOS在鉴别诊断兰州地区3~6岁儿童哮喘、过敏性鼻炎、下呼吸道感染方面展现出较好的潜力,且基于三者构建的多因素逻辑回归模型可有效提升对哮喘的诊断准确性。

基于倾向评分回归调整探讨艾灸治疗常年变应性鼻炎灸感与灸效的前瞻性队列研究

目的:采用倾向评分回归调整(PSRA)均衡基线资料,比较艾灸同一穴位(上印堂)不同灸感(热敏灸态vs静息态)治疗常年变应性鼻炎(PAR)灸效差异。方法:采用前瞻性队列研究设计,纳入106例合格受试者,用艾条悬灸上印堂激发灸感,根据热敏灸感的有无,自然分组为热敏灸组(n=69)和传统灸组(n=37)。两组均在上印堂施灸,每次时间为30min,每天1次,连续四周,共计24次。结局指标为变异性鼻炎症状量表(TNSS)、伴随症状量表(TNNSS)以及生存质量调查问卷(RQLQ)。结果:使用PSM进行1:1配对,匹配容差0.02,成功匹配25对,共50例患者。两组患者基线资料均无统计学差异(P>0.05)。匹配后,治疗后热敏态组TNSS评分为4.01±1.52,低于静态组的5.42±1.10,有统计学意义(P<0.01);热敏态组TNNSS评分为1.26±0.43,低于静态组的1.52±0.50,有统计学意义(P<0.01);热敏态组RQLQ评分为37.62±6.01,低于静态组的47.22±7.76,有统计学意义(P<0.01);说明热敏灸组在改善PAR患者症状、体征和生活质量方面均有疗效优势。将倾向性评分匹配前单因素分析具有差异的结果纳入进行多因素logistic回归分析,结果显示,是否用药、病程为艾灸疗效的独立危险因素(P<0.05)。结论:艾灸热敏态穴位治疗PAR的临床疗效优于静息态组。

《变应性鼻炎及其对哮喘的影响》指南2010年修订版解读

《变应性鼻炎及其对哮喘的影响》(Allergic rhinitis and its impact on asthma,ARIA)指南最新修订版(简称ARIA2010版)最近由《Journal of Allergy and Clinical Immunology》杂志全文发表。与旧版ARIA指南使用循证医学的方法不同,ARIA2010版采用推荐、评估、发展和评价分级(Grading of Recommendations,Assessment,Development and Evaluation,GRADE)工作组提出的透明的、系统化的方法修订了临床建议,以提高其实用价值。

腺样体切除术后圆枕增生与变应性鼻炎的关系

人们已经注意到,变应性鼻炎（allergic rhinitis,AR）患儿腺样体肥大发病率较非AR患儿高,近几年AR患儿发病率逐渐增高,我们考虑,腺样体切除术后圆枕增生可能和AR有密切关系,本文用回顾性分析的方法对此问题进行了探讨。

南昌地区变应性鼻炎患者变应原皮肤点刺试验结果分析

由于生活习惯、地理环境和气候的不同,各地变应性鼻炎（AR）变应原分布特点有很大区别,了解和分析南昌地区常见变应原分布情况,对预防和治疗该地区AR具有重要的指导意义。我科对南昌地区959例AR患者进行变应原皮肤点刺试验（skin prick test,SPT）,现报道如下。

鼻腔细胞学研究现状

鼻腔细胞学是一种安全、便捷、高效的检测方法,是指导鼻腔疾病诊断和治疗的极好指标,在鼻科学病理研究中占有重要地位。鼻腔细胞的收集方法包括擤鼻法、鼻黏膜刷片法、鼻黏膜拭子法、鼻黏膜刮片法、鼻腔盥洗液法、膨胀海绵法和鼻黏膜活检法。鼻分泌物中炎性细胞的检出是鼻腔、鼻窦炎性疾病诊断和鉴别的重要依据。炎性细胞和鼻黏膜上皮结构细胞的改变还可作为评价鼻腔炎症疗效的客观指标。鼻腔细胞学研究还用于鼻腔、鼻窦炎性疾病的疗效预测和鼻腔微生物学研究。然而诊断标准不统一、技术手段相对简单的现状局限了鼻腔细胞学的发展,这一领域技术的进步将促进人们对鼻部疾病认识的深入。

重视鼻炎研究

鼻炎（rhinitis）和（鼻）鼻窦炎（sinusitis或rhinosinusitis）是耳鼻咽喉头颈外科的主要常见病,美国（人口3亿）每年诊治过敏性鼻炎患者约3～6千万例（人口10%～20%）,鼻窦炎患者约为3.1千万例（人口10%）。鼻炎是由鼻腔黏膜炎症反应而导致的急性或慢性症状性疾病,慢性鼻炎更为常见。