| Part 1.The structural characteristics and influence factors in nomal bronchus and alveolar using confocal laser endomicroscopyBackground:Histopathological examination is the gold standard to evaluate whether the bronchoalveolar structure is abnormal or not,bronchoscopic probe-based confocal laser endomicroscopy(p CLE)is a real-time imaging technique for pulmonary autofluorescent elastic fibre and leads to high-resolution imaging at the cell level in vivo,and foreign researchers have used p CLE to observe the airway subepithelial basement membrane,alveolar elastic fibers,and microvascular structures.However,it is still unclear that the confocal imaging characteristics of the normal bronchial alveolar elastic fibers composed of different age in vivo and their correlation with ageing.Besides,the relationship between the elastic fiber autofluorescence density and the changes in alveolar structure is not yet clear.Objective:1.To define the confocal imaging characteristics of normal bronchoalveolar structures;2.To explore the value of CLE measurement of elastic fiber fluorescence density to evaluate the destructive degree of alveolar structures.Subjects and Methods:subject and inclusion criteria: All subjects had no other basic disease 、normal pulmonary function and chest CT showed peripheral pulmonary nodules.Methods: The patient underwent bronchoscopy under awake sedation,and was sent into the p CLE probe through the bronchoscope working channel to explore the anterior,external,and posterior basal segments of the double lower lungs respectively.We measured the bronchial wall autofluorescence intensity 、alveolar elastic fibre autofluorescence intensity 、 alveolar elastic fibre thickness 、 alveolar elastic fibre extension length increase percentage(AL%)、 microvessel wall autofluorescence intensity and evaluate alveolar elastic fibre pattern.Results:A total of 82 patients were enrolled,included 27 males and 55 females,aged 18-79 years,and 503 lung segments were analyzed.(1)Confocal imaging characteristics of normal bronchoalveolar structure: the elastic fibers of the bronchi are arranged in parallel and regular;the round alveoli are cross-linked and distributed in a "trapezoidal suture";both the autofluorescence density and thickness of the alveolar opening elastic fibers are significantly higher than the distal;destructiveness changes in alveolar elastic fibers can be observed in normal subject.(2)Correlation between bronchoalveolar structure and age: alveolar opening elastic fibre autoflurescence intensity and microvessel wall intensity correlated positively with age(r=0.642 and r=0.637,all P<0.01);AL% correlated negatively with age and elastic fibre autoflurescence intensity(r=-0.458 and r=-0.300,all P<0.001).(3)Difference of bronchoalveolar structure in different age group: In the aged 18-39 group、40-54 group、55-79 group,the mean ±SD autoflurescence intensity of the bronchial wall were significantly increased(8.17±2.88 v.s.11.54±3.49 v.s.14.50±4.18,all P<0.05);the mean ±SD autoflurescence intensity of the alveolar opening elastic fibre were significantly increased(11.48±4.19 v.s.19.23±4.84 v.s.26.10±11.08,all P<0.05);the mean ±SD autoflurescence intensity of the microvessel wall were significantly increased(11.01±3.96 v.s.19.14±6.10 v.s.24.82±11.22,all P<0.05);AL% were decreased(81.85±51.31 v.s.44.54±31.79 v.s.35.96±23.88),and 18-39 group was significantly larger than 40-54 group and 55-79 group,respectively(all P<0.05);the proportion of alveolar elastic fibre appear ruptured were increased(6.94% v.s.13.44% v.s.20.81%),and 55-79 group was significantly higher than 18-39 group and 40-54 group,respectively(all P<0.05).(4)Relationship between alveolar elastic fibre autofluorescence intensity and alveolar elastic fibre appear ruptured: The proportion of alveolar elastic fibre appear ruptured in the autoflurescence intensity of elastic fibre 5-14group、15-21group、22-75 group were increased(7.32% v.s.13.81% v.s.21.52%),and 22-75 group was higher significantly than 5-14 group and 15-21 group respectively(all P<0.05).Conclusion:With the increase of age,the autofluorescence density of alveolar elastic fibers and microvessel walls gradually increase;with the increase of age,the proportion of abnormal changes in elastic fibers significantly increase;with the increase of autofluorescence density,the proportion destructiveness of elastic fibers also significantly increase.Increased levels of autofluorescence density may indicate the destructivess of alveolar elastic fibers.Part 2.Analysis of the alveolar structure imaging characteristics in smoker and chronic obstructive pulmonary disease using confocal laser endomicroscopyBackground:Bronchoscopic probe-based confocal laser endomicroscopy(p CLE)is a real-time imaging technique for evaluating alveolar elastic fiber and macrophages containing tobacco tar.Studies assessed the alveolar macrophages infiltration and the relationship between alveolar morphologic index of chronic obstructive pulmonary disease(COPD)and CT as well as lung function.However,it is still unclear that confocal imaging characteristics of the bronchoalveolar structure and macrophages between current smokers,ex-smokers and chronic obstructive pulmonary disease and their correlation with smoking history.Besides,CLE assessment of the degree of structural damage of alveolar elastic fibers in COPD patients need further exploration.Objectives:1.To explore the relationship between the confocal imaging characteristics of bronchoalveolar structures in smoking subject.2.Compare the confocal imaging characteristics of alveolar structure in COPD patients with vary degrees of severity.Subjects and Methods:Subjects and inclusion criteria: All smokers subjects had no other basic disease 、normal pulmonary function and chest CT showed peripheral pulmonary nodules;patients diagnosed with COPD.Methods: The patient underwent bronchoscopy under awake sedation,and was sent into the p CLE probe through the bronchoscope working channel to explore the anterior,external,and posterior basal segments of the double lower lungs respectively.Results:We recruited 119 subjects from The First Affiliated Hospital of Guangzhou Medical University between Jan 2017 and Dec 2019.The study cohort consisted of 54 smokers(35 current smokers、6 ex-smokers who had stopped smoking for less than 1 year、13 ex-smokers who had stopped smoking for more than 1 years)、29 COPD patitents(stage I,n=17;stage II-III,n=12)and 36 control subjects who were agematched with smokers and COPD.(1)Relationship between imaging characteristics of alveolar structure and smoking status: the alveolar elastic fibre autofluorescence intensity and the microvessel wall autofluorescence density significantly decreased in the current smoker group、ex-smoker group and non-smoker group,(74.20 ± 30.60 v.s.40.88 ± 23.14 v.s.27.09 ± 12.24,all P <0.05;69.87 ± 24.25 v.s.33.07 ± 17.42 v.s.27.39 ± 10.72,all P <0.05).The proportion of elastic fibre appeared hyperplasia significantly decreased in the current smoker group、ex-smoker group and non-smoker group,(63.01% v.s.46.05% v.s.3.74%,all P <0.05).(2)Relationship between imaging characteristics of alveolar structure and macrophage and smoking history: in current smoking subjects,the alveolar elastic fibre autofluorescence density was positively correlated with the amount of cigarettes smoked per day(r=0.282,P<0.001);the microvessel wall autofluorescence density was positively correlated with the amount of cigarettes smoked per day(r=0.358,P=0.003);the number of autofluorescent macropgages per field of view was positively correlated with the amount of cigarettes smoked per day(r=0.364,P=0.032);the number of autofluorescent macrophages per field of view was correlated with the autoflurescence intensity of macrophages(r=0.602,P<0.001).(3)Difference in the morphologic change between non-smoker group 、smoker group and COPD group: the proportion of elastic fibre appear ruptured significantly increased in the non-smoker group、smoker group and COPD group,(3.74% v.s.46.05% v.s.63.01%,all P <0.05).Conclusions:The alveolar elastic fibers autofluorescence density is closely related to smoking status,and it increases significantly in the non-smoker,ex-smoker and current smoker,among which the cigarettes smoked per day increases in current smoker.The number of alveolar macrophages in current smokers is positively correlated with the cigarettes smoked per day.The proportion of alveolar elastic fiber appear ruputed significantly increase in non-smoker 、smoker and COPD.p CLE can find that the destruction degree of alveolar elastic fibers in smokers with normal lung function is significantly higher than that in non-smoker. |
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