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Establishment of Eosinophilic Bronchitis Mouse Model

Establishment of Eosinophilic Bronchitis Mouse Model

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2016中华医学会呼吸病学年会暨第十七次全国呼吸病学学术会议

作者： ChenhuiLi MinPeng LiyanChen The 1st Affiliated Hospital of Guangzhou Medical University

目的 Objects：Eosinophilic Bronchitis(EB)is a kind of disease that has the characters with chronic cough,eosinophilic airway inflammation and effective treatment of steroid but has no airway hyperresponsiveness(AHR).But now no animal model has been acknowledged through the *** EB model is emergent to be established to explore the mechanism,outcome and treatment of EB.方法 Methods：Forty Balb/c female mice(ten weeks)were divided into four groups：NS(normal saline),AS(asthma),EB,DXM(Dexamethasone).Asthma mice were challenged intranasally with 200ug OVA,at the same time EB and DXM mice were challenged with 10ug *** DXM mice were treated with DXM 5mg/kg i.p before being challedged and airway responsiveness ***,NS control mice received normal *** completing model-creation,cough reflex sensitivity and airway responsiveness were detected in the four ***,the Eosinophil Cell Counts(EOS)in bronchoalveolar lavage fluid(BALF)was detected after exposure to increasing doses of methacholine(MCh).Lung histological sections were examined for inflammatory infiltration.结果 Results：Cough evoked by aerosolized capsaicin was significantly increased by challenged with 10ug and 200ug OVA(Figure1A,B,p＜0.01)in EB and AS *** in DXM group,cough can be significantly improved,compared with AS and EB group(Figure1C,p＜0.01).In AS group,lung resistance(RL)was significantly increased at Mch 12.5mg/ml and 25mg/ml compared with other three ***,RI in the EB group was not significantly different compared with the NS and DXM group at any MCh concentration(Figure1D,p＜0.05).Eosinophils in BALF were increased significantly in EB and AS groups compared with the NS group,but no significant differences were observed between the two *** the contrast,in the DXM group eosinophils were decreased significantly in BALF,compared with EB and AS(Figure1E,p＜0.05).Furthermore,Inflammatory cells were seen around bronchioles in EB group but can be improve

关键词： Cough Eosinophilic Bronchitis

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Caveolin-1 deficiency causes pulmonary fibrosis alters energy metabolism and induces endoplasmic reticulum stress in the alveolar epithelium

Caveolin-1 deficiency causes pulmonary fibrosis alters energ...

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2016中华医学会呼吸病学年会暨第十七次全国呼吸病学学术会议

作者： YingZhu Center for Translational Medicine Thomas Jefferson University

目的 Caveolin-1 (Cav-1), a major structural protein of caveolae, is involved in many physiological and pathophysiological process such as energy homeostasis and lung *** it is well-recognized that Cav-1 deficiency promotes pulmonary fibrosis in mice, the relationship to lung energy metabolism remains unexplored. 方法 Cav-1 gene disrupted mice (Cav-1-/- mice) and the corresponding controls B6129 mice (male, 8-10 weeks old) were used. When applicable, mice received 12.5 mg of glucose metabolism inhibitor 2-deoxyglucose (2-DG) or vehicle in a total volume of 100 μl of sterile PBS by daily ip *** activation and the unfolded protein response were assessed in whole lung tissue and isolated primary AECII cells by performing Western blot *** levels were measured using commercially available kits. Lung fibrosis was assessed by measuring TGF-β1 levels in BAL fluid, by quantifying transcripts and protein levels for TGF-β1 and collagen1α*** cell death was assessed in whole tissue by evaluating caspase 3 *** epithelial A549 cells were transfected with Cav-1 shRNA lentivirus and used for evaluating Cav-1 depletion-induced metabolic and ER tress in culture. 结果 We found that Cav-1 deficiency induced severe metabolic stress in the lung, significantly suppressing ATP levels and enhancing the phosphorylation of ***, we showed that alterations in metabolic homeostasis were associated with an increase in ER stress as evident by an upregulation in each of the major ER stress sensors (IRE-1, ATF6, PERK) in whole tissues and primary alveolar type Ⅱ cells from Cav-1 -/- ***, we found that metabolic and ER stress was enhanced in the lungs of Cav-1 -/- mice undergoing further metabolic with 2-DG ***, this coincided with the increase in TGF-β1 and collagen levels and apoptotic ***, silencing Cav-1 with shRNA induced AMPK activation, ER stress and caspase-3 activation in A549 cells. 结论 Our data provide

关键词： endoplasmic reticulum stress Caveolin-1 pulmonary fibrosis energy metabolism

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Does Norepinephrine Participated In The Central Regulation Of Genioglossus During CIH In Rats?

Does Norepinephrine Participated In The Central Regulation O...

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2016中华医学会呼吸病学年会暨第十七次全国呼吸病学学术会议

作者： LingZhou WeiWang AiDiWang JianKang THE FIRST HOSPITAL OF CHINA MEDICAL UNIVERSITY

目的 Upper airway dilator muscles(e.g.,genioglossus) play an important role in maintaining the patency of upper *** sleep apnea syndrome (OSAS) is characterized by recurrent upper airway obstruction and chronic intermittent hypoxia (CIH) during sleep,but it does not occur during *** study showed that CIH could induce the increase of density of noradrenergic terminals and α1-adrenoceptor immunoreactivity in the hypoglossal (Ⅻ) nucleus of the ***'s still unknown whether norepinephrine is involved in the central regulation of genioglossus during CIH.方法 32 male Wistar rats were randomly divided into NC group、CIH group、CIH with drug group(the α1-adrenoceptor antagonist prazosin,0.2 mM, 60 nl) and CIH with Nacl group(0.9％,60nl).The hypoxic compartment (A-15274-PEVAC,BioSpherix,NY) was used to set up CIH *** CIH exposure lasted *** oxygen concentration levels oscillated between 21％ and 10％ with 188-s period for 8 h/day (8 AM-4 PM).At the same time,NC group rats were exposed to the alternating cycles of air under identical experimental conditions as CIH in *** rat brain stereotaxic apparatus (Life Science Co., *** Rui Wode) was used to locate the hypoglossal nucleus of all *** drug and Nacl were microinjected to the hypoglossal nucleus by catheter series(Life Science Co., *** Rui Wode).Transcranial magnetic stimulation(TMS) was applied for every rat to record genioglossus movement cortex reaction on the 1st,7th,14th,21st day of the experiment.结果 Comepared to the air group,the CIH group showed shortened latencies (P＜0.05).For those rats exposed to CIH, CIH with drug group showed the tendency of prolonged latency comepared to the other two groups,but the difference was significant only on the 14th day(P＜0.05).结论 These results indicated that norepinephrine participated in the central regulation of genioglossus during CIH in rats,especially on the 14th day.

关键词： Genioglossus norepinephrine chronic intermittent hypoxia hypoglossal nucleus

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The Role of Extracellular Heat Shock Protein 90 Alpha on Pulmonary Fibrosis

The Role of Extracellular Heat Shock Protein 90 Alpha on Pul...

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2016中华医学会呼吸病学年会暨第十七次全国呼吸病学学术会议

作者： HangmingDong LishanLuo ChaowenHuang HaijinZhao ShaoxiCai Nanfang Hospital Southern Medical University

目的 Pulmonary fibrosis is a progressive disease with poor prognosis and *** is characterized by lung fibroblast actvation and extracellular matrix(ECM) deposition,.Heat shock protein 90(Hsp90) plays a significant role in multiple cellular process, especially extracellular heat shock protein 90 alpha(eHsp90α) can promotes wound healing induced by fibroblast actvation and migration.方法 Recombinant TGF-β1 was used to induce human lung fibroblast activation and ECM *** protein level of intracellular Hsp90α and Hsp90β was detected by western *** condition media of lung fibroblast was collected to detected the expression of extracellular Hsp90α and Hsp90β.Anti-Hsp90 monoclonal antibody (mAb) was applied at 50ug/*** fibroblast activation and ECM production were respectively evaluated by expression of alpha smooth muscle actin(α-SMA), fibronectin and collagen Ⅰ.C57/BL mice modle of pulmonary fibrosis was induced by 3U/kg bleomycin (BLM).eHsp90α in BALF was determined by *** fibrosis was assesssed by HE staining, masson-trichrome staining, the content of ***, the expression of Hsp90α, α-SMA, collagen Ⅰ was determined by immunohistochemistry.结果 Secretion of Hsp90α and Hsp90β by human lung fibroblast , as well as the expression of intracellular Hsp90α and Hsp90β, is significantly increased after stimulation by TGF-β1 .Anti-Hsp90α mAb prevents lung fibroblast activation and ECM synthesis, shown by decreased expression of α-SMA, fibronectin and collagen Ⅰ.Moreover, eHsp90α is greatly upregulated in BALF of BLMinduced fibrotic ***90α is increased in BLM-induced fibrobtic ***-Hsp90α mAb can alleviate pulmonary fibrosis induced by BLM indicated by decreased ECM deposition and downregulated expression of α-SMA,collagen Ⅰ.结论 eHsp90α promotes lung fibroblast activation and ECM production, therefore plays an important role in the development of pulmonary ***-Hsp90α monoclonal antibody significantly ameliorat

关键词： Pulmonary Fibrosis HSP90 TGF-β1 monoclonal antibody

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A H2O2 induced mouse model of cough hypersensitivity via oxidative stress

A H2O2 induced mouse model of cough hypersensitivity via oxi...

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2016中华医学会呼吸病学年会暨第十七次全国呼吸病学学术会议

作者： LiQiYang ShanZhong ChuQinHuang KeFangLai TheFirstAffiliatedHospitalofGuangzhouMedicalUniversity GuangzhouInstituteofRespiratoryDis TheFirstAffiliatedHospitalofGuangzhouMedicalUniversity

目的 Long-term inhalation of cigarette smoke or air pollutants can both result in the cough hypersensitivity. Many researchers reported that amount of reactive oxygen species were consisted in cigarette smoke and air pollutants. Present study aims to investigate the relationship between airway oxidative stress induced by H2O2 and cough sensitivity, and to establish an appropriate nebulizing concentration of H2O2 and nebulizin exposure time for the modeling. 方法 SPF grade Balb/c were randomly grouped according to nebulizating concentration(0.2％, 1％, 5％) and exposure time(3 d, 7 d, 10 d, 14 d). Another 3 n-acetylcysteine(NAC)intragastrical treatment groups would be seted up in each concentration corresponding to the model control group which present the highest frequency in cough times. Mice were exposed in H2O2 via nebulizer in a chamber. Each nebulization remains for 20min twice a day. NAC intragastrically administration were manipulated between two nebulization. A capsicin cough challenge was manipulated in the day after termination of nebulization. After cough challenge, animals were sacrificed. Bronchoalveolar Lavage Fluid (BALF) were gathered for total cell counts and differential leukocyte counts. Lung tissues were also gathered for real time PCR for Transient receptor potential vanilloid 1(TRPV1), Transient receptor potential ankyrin 1(TRPA1) mRNA expressions and detecting superoxide dismutase (SOD) activity, glutathione peroxidase (GSH-px) activity. 2 mice in each groups were selected for trachea and lung tissues histological evaluations.

关键词： chronic cough H2O2 oxidative stress TRPV1 TRPA1 animal model

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Treatment with YKL-40siRNA down-regulates the expression of eotaxin,IL-5,GM-CSF in OVA-stimulated airway epithelial cells

Treatment with YKL-40siRNA down-regulates the expression of ...

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2016中华医学会呼吸病学年会暨第十七次全国呼吸病学学术会议

作者： SuqinBen Department of Respiratory Medicine Shanghai General HospitalShanghai Jiao Tong UniversityShanghaiChina

目的 YKL-40 is highly expressed in airway inflammation and remodeling,and higher levels of YKL-40 are detected in patients with inflammatory ***-40 may participate in the mechanism of inflammatory mediators that regulate eosinophil-mediated airway inflammation and contribute to the pathogenesis of *** the epithelium,YKL-40 could modulate the expression of eotaxin,granulocyte-macrophage colony-stimulating factor(GM-CSF),and interleukin(IL)-5,and the latter played an important role in eosinophilic airway ***,YKL-40 could be an attractive target in the design of asthma therapies.方法 Mouse airway epithelial cells were isolated and identified by immunofluorescent *** primary mouse tracheal epithelial cells were cultured with OVA for 48 h and transfected with YKL-40 *** relative levels of YKL-40,IL-5,GM-CSF,and Eotaxin mRNA transcripts were determined by semi-quantitative RT-PCR,and their proteins were determined by ELISA and Western blot assays,respectively.结果 The levels of YKL-40 protein in cells transfected with siRNA-YKL-40(YKL-40-si)were decreased by 70％compared with the levels in primary mouse tracheal epithelial cells transfected with siRNA-negative control(YKL-40-NC)or non-transfected epithelial cells(YKL-40-0)by Western *** levels of eotaxin,GM-CSF,IL-5 mRNA and protein in cells transfected with siRNA-YKL-40(YKL-40-si)were markedly decreased compared with those in cells transfected with siRNA-negative control(YKL-40-NC)or non-transfected epithelial cells(YKL-40-0).结论 YKL-40 siRNA down-regulates the expression of eotaxin,IL-5,GM-CSF in OVA-stimulated airway epithelial *** finding could extend the prospective use of YKL-40 siRNA in asthma research and enlarge the armamentarium for treating asthma.

关键词： asthma eotaxin epithelium GM-CSF IL-5 YKL-40

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Antioxidant Tempol ameliorates intermittent hypoxia-induced melanoma lung metastasis in a mouse model of OSA

Antioxidant Tempol ameliorates intermittent hypoxia-induced ...

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2016中华医学会呼吸病学年会暨第十七次全国呼吸病学学术会议

作者： LianLi JieCao BaoyuanChen Respiratory Department Tianjin Medical University General Hospital

目的 Obstructive sleep apnea (OSA) has been recently associated with tumor incidence and cancer-related *** hypoxia (IH) mimicking OSA contributes to tumor growth and metastasis in a mouse ***, the study in our lab has found that oxidative stress and inflammatory responses could mediate IH-induced melanoma lung metastasis in a mouse model of OSA (Li et al, unpublished data).Here we blocked oxidative stess with antioxidant tempol to explore the tumor lung metastasis induced by OSA-like IH.方法 C57BL/6J male mice (8-week-old) were injected with B16F10 melanoma cells via tail *** these animals were randomly divided into three groups (n = 8 each)：nomoxia group, OSA-like IH group,and tempol group (exposed to IH and treated with tempol).Number of lung metastatic colonies was assessed at 21 days after exposure to *** tissues were harvested to detect the markers for oxidative stress(NF-E2-related factor 2, NRF2) and inflammation (nuclear factor kappa B P65, NF-κB P65) by western blotting.结果 The number of melanoma lung metastasis in OSA-like IH group was significantly higher than that in nomoxia ***, this number in tempol group was significantly lower than that in IH *** protein levels of NRF2 and NF-κB P65 in OSA-like IH group were significantly higher than that in nomoxia ***, these levels in tempol group was significantly lower than that in IH group.结论 These results suggest that antioxidant tempol is, at least partially, capable of blocking tumor metastasis to the lung induced by OSA-like *** intervention may be useful as the feasible strategy to reduce the impact of OSA on cancer modulation.

关键词： Tempol intermittent hypoxia lung metastasis OSA

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Inhaled corticosteroids improve lung function,airway hyperresponsiveness and airway inflammation but not symptom control in mild intermittent patients:systemic review and meta-analysis

Inhaled corticosteroids improve lung function,airway hyperre...

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2016中华医学会呼吸病学年会暨第十七次全国呼吸病学学术会议

作者： WeiDu LingZhou YingmengNi YuanyuanYu FangWu GuochaoShi Ruijin Hospital Shanghai Jiao Tong University School of Medicine

目的 It is controversial whether ICS(inhaled corticosteroid)should be used in intermittent asthma patients,and so we are aimed to assess the effect of ICS on these patients compared with placebo or other therapies.方法 Medline,Embase and CNKI database updated to November 2015 were searched and meta-analysis was conducted.结果 In adult patients,when compared with placebo,ICS increased FEV1(WMD 4.01％,95％CI 2.74～5.27％)and alleviated airway hyper-responsiveness(indicated as log transformed PC20FEV1,SMD 0.98,95％CI 0.64～1.32).ICS also reduced sputum eosinophil percentage(WMD-1.3％,95％CI-2.11～-0.49％)and FeNO level(WMD-12.71ppb,95％CI-16.21～-9.21ppb).But the symptom score did not change after ICS treatment(SMD-0.13,95％CI-0.26～-0).When compared with LTRA(leukotriene receptor antagonists),ICS had no advantage in increasing FEV1(WMD-0.04L,95％CI-0.29～0.21L)or reducing sputum eosinophil percentage(WMD-6％,95％CI-12.38～0.38％)or symptom score(SMD 0.44,95％CI-0.02～0.9).However,in child patients,ICS significantly increased the possibility of symptom control when compared with placebo(RR 8,95％CI 1.04～61.52)or LTRA(RR 2.67,95％CI 0.39～18.42).结论 ICS can improve lung function and alleviate airway hyper-responsiveness and airway inflammation but cannot influence symptom score,and has no advantage over LTRA on lung function improvement and airway inflammation control in adult patients with mild intermittent *** in child patients,the benefit of ICS in symptom control is more obvious than LTRA.

关键词： asthma ICS meta-analysis inflammation respiratory function test clinical respiratory medicine

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Screening and managing obstructive sleep apnoea in nocturnal heart block patients: An observational study

Screening and managing obstructive sleep apnoea in nocturnal...

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2016中华医学会呼吸病学年会暨第十七次全国呼吸病学学术会议

作者： XuWu ZilongLiu SuchiChang CuipingFu ShanqunLi Department of Pulmonary Medicine Center of snoring and Sleep Apnea MedicineZhongshan Hospital of Fudan UniversityShanghai200032China

目的 Nocturnal heart block often occurs in patients with obstructive sleep apnoea (OSA).It is more likely to be undiagnosed in heart block patients who are ignorant of the symptoms of sleep *** Questionnaire (BQ) is a highly reliable way to discover the risk factors of OSA, whereas the validity in sleep-related heart block patients is *** performed an observational study to address these issues and confirmed the potential protective effect of continuous positive airway pressure (CPAP).方法 Patients who were previously diagnosed with nocturnal heart block with R-R pauses exceeding 2 seconds were retrospective screened from the ECG centre of Zhongshan *** recruited participants completed Berlin Questionnaire and underwent polysomnography synchronously with 24-hour Holter monitoring.A cross-sectional analysis was performed to confirm the association between nocturnal arrhythmia and OSA, as well as to assess the diagnostic accuracy of the ***, subjects diagnosed with OSA (apnoea-hypopnoea index ＞ 5) underwent 3 consecutive days of CPAP *** the third day, patients repeated 24-hour Holter monitoring within the institution of CPAP.结果 The symptoms of disruptive snoring and hypersomnolence in 72 enrolled patients were more related to the occurrence of nocturnal heart block (r = 0.306, 0.226, respectively, p = 0.015, 0.019) than syncope (r = 0.134, p = 0.282) and palpitations (r = 0.106, p = 0.119), which were prominent trait of our study *** sensitivity, specificity, positive and negative predictive value of the BQ at a cut-off point of 5 of AHI for detecting OSA in heart block patients was 81.0％, 44.4％, 91.07％ and 25％.Nocturnal heart block does not appear to occur exclusively in severe sleep *** frequent occurrence of arrhythmias in prominent oxygen desaturation supports the correlation between *** therapy resulted in significant decrease in the average number of episodes of heart block, from 148.58 ± 37

关键词： Obstructive sleep apnoea Heart block Berlin questionnaire Continuous positive airway pressure

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Diagnostic accuracy of the Berlin questionnaire in detecting OSA in patients with glucose metabolic dysfunction and Its Interference with CPAP

Diagnostic accuracy of the Berlin questionnaire in detecting...

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2016中华医学会呼吸病学年会暨第十七次全国呼吸病学学术会议

作者： SuchiChang ZilongLiu XuWu ShanqunLi Department of Pulmonary Medicine Center of snoring and Sleep Apnea MedicineZhongshan Hospital of Fudan University

目的 Obstructive sleep apnea (OSA) is associated with alterations in glucose *** OSA may also deteriorate *** Questionnaire (BQ) is effective to identify patients at high risk for OSA, however, its sensitivity and specificity in patients with glucose metabolic dysfunction remain *** continuous positive airway pressure (CPAP) can act as an adjunct in diabetes control or insulin sensitivity needs to be determined.方法 121 patients with OSA symptoms were recruited for polysomnogram (PSG) and asked to complete Berlin Questionnaire (BQ), Pittsburgh Questionnaire (PSQI), and Epworth Sleepiness Scale (ESS).Patients were then scheduled the following morning for a 75 g oral glucose tolerance test (OGTT).According to the glycemic parameters, patients were divided into control, impaired fasting glucose (IFG), impaired glucose tolerance (IGT), and type 2 diabetes mellitus (T2DM) group for subgroup *** received CPAP treatment for 3 months, and were then recruited for *** analysis with SPSS 22.0 was performed to study the diagnostic accuracy of BQ and the efficacy of CPAP therapy.结果 The sensitivity, specificity and accuracy of BQ in diagnosis of OSA with glucose metabolic dysfunction when AHI cut-off values are set at 5, 15, and 30, were： 72.29％, 100％, 72.62％;76.92％, 100％, 78.57％;76.92％, 47.37％, 70.24％, respectively.3 months of CPAP therapy effectively reduced the clinical symptoms and PSG parameters for all three groups, simultaneously improving OGTT results and HOMA-IR (P＜0.05).The improvement in glucose tolerance was more significant in the diabetes type Ⅱ group, with statistical significance (P＜0.05) for OGTT glucose levels, glycosylated hemoglobin, and decrease in HOMA-IR.结论 BQ is highly accurate in screening OSA patients with glucose metabolic dysfunction and can be considered an effective screening *** investigation of BQ in the diagnosis and management of mild to moderate OSAHS in patients with gluco

关键词： Obstructive sleep apnea Berlin Questionnaire glucose metabolic dysfunction CPAP

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