The Clinical Analysis In Patients Of Chronic Obstructive Pulmonary Disease With Invasive Pulmonary Aspergillosis And Its Animal Model Establishment

Objective:1.By collecting the data about 24 cases of chronic obstructive pulmonary disease(COPD)with invasive pulmonary aspergillosis(IPA),we summarize and analyze their clinical characteristics,thus provide help for early diagnosis andtreatment.2.To explore the establishment method of animal models and further study the pathogenesis of COPD combined with IPA ultimately.Methods:1.Collect the clinical data of patients who were hospitalized and treated in the Second Hospital of Jilin University pneumology department from December 2010 to September 2016,and of whom 24 cases were COPD combined with IPA as well as 24 cases were COPD alone.The data contain:(1)general information and related medical history:age,gender,smoking index,underlying disease,the times formerly hospitalized and the application of glucocorticoid and broad-spectrum antibiotics;(2)clinical manifestation and related treatment:symptoms,signs,imaging manifestations about chest,the outcome of bronchoscope,laboratory inspection,results of pathogenic detection,treatment received after admission,the length of stay and condition of recovery.Make a retrospective statistical analysis of the clinical data above-mentioned.2.The establishment of animal models of COPD combined with IPA:select 16 wild type(WT)C57BL/6 mice,then divide the mice into 2 groups as normal control(8)and experimental group(8).The mice in experimental group were exposed Nose-only inhalation system,4 times a day,lasting 30 minutes every time,15 minutes interval,and the disposition continues 16 weeks;thenceforward the mice were given methylprednisolone in a dose of 100mg/kg every day,lasting 5 days.Next day the mice were offered the spore suspension liquid 30 ul in a concentration of 1×106CFU/ml in a tracheal puncture method,followed by being kept a state of erecting for 3-5 minutes and being given a disposition of suturing neck skin.After successful vaccination,the mice were given vancomycin in a dose of 15mg/kg every day,ceftazidime in a dose of150mg/kg every day,gentamicin in a dose of 5mg/kg once every other day in order to preventing mixed infection.The mice of normal control group were not exposed Nose-only inhalation system,and inhale air in the glass cabinet.After that the mice were given same volume saline instead of methylprednisolone and the spore suspension liquid,and other dispositions were the same with the mice in experimental group.Watch the general condition including mental state,activity and eating situation,and weigh all the mice every 2 weeks.After being infected successfully for72 hours,all the mice were rendered united disposal covering measuring weight,testing mice's pulmonary function test,etiologically culturing the lung tissue and pathological examination.Results:1.clinical data analysis(1)Patients of two group have no significant difference in the aspects of age and gender(P>0.05).(2)Patients of COPD combined with IPA are apparently higher than patients of COPD alone in the fields of smoking index and average hospitalization time(P <0.05).(3)Analyzing the general data of two groups,patients of COPD combined with IPA are more common than patients of COPD alone at the aspects of oral or intravenous hormonotherapy greater than or equal to 7 days,applying broad-spectrum antibiotics greater than or equal to 14 days,admission times in 1 year greater than orequal to 2 times,hospitalized in ICU before diagnosis,the level of albumin less than32g/L and hemoglobin less than 115g/L(P<0.05).About the complications,patients of two groups are no statistical difference in the fields of combining hypertension,coronary heart disease,diabetes(P>0.05).(4)Patients of two groups both exist the symptom of cough,and other symptoms have no obvious difference containing expectoration,blood-stained sputum,fever,dyspnea,chest pain,Rhonchi and Moist crackles(P>0.05).(5)Comparing the total white blood cells and neutrophilic granulocyte percentage,the patients' of COPD combined with IPA are both greater than the patients' of the COPD alone,11.072±6.10 vs 7.829±2.75 and 78.758±14.77 vs65.308±12.34,which is a significant statistical difference(P<0.05).(6)Positive incidence of related laboratory inspections and imaginational examinations in the group of COPD combined with IPA are described as follows:GM test(54.17%),G test(83.33 %),sputum culturing(87.5%),19 cases aspergillus fumigabtus cultured positive(79.17%),3 cases aspergillus niger positive(12.5%)and2 cases aspergillus flavus positive(8.3%)who all belonged to the patients whose sputum culturings were cultivated and had strains grow,patch effusion shadows in imaginational examinations(66.67%),nodular shadows(containing halo sign)(34.78%),pulmonary segments and lobe atelectasis or consolidation shadows(17.39%),single or multiple hollow(17.39%),two lungs ground-glass attenuation(13.04%),air crescent sign(4.34%),pleural effusion(34.78%).(7)The treatment of group of COPD combined with IPA:combine antifungal therapy with broad-spectrum antibiotics,and analyze initially the treatment situation and effect of resistance to aspergillus,then the consequence shows that applying separately itraconazole,voriconazole,amphotericin B,caspofungin lonely and combined therapy have no significant difference(P> 0.05).Eventually 17 cases(70.83%)improved and discharged,and 7 cases died including 4 deaths confirmed within hospital and 3 deaths in the follow-up after discharging from hospital.2.The establishment of animal models of COPD combined with IPA(1)The change of weight:From the fourth week on,the experimental group mice ' s weight happened to appear apparent diminution compared to the normal control'(P<0.05).(2)Lung function: the total airway resistance(R),airway resistance(Rn)and respiratory system elastic resistance(E)of experimental group mice remarkably increased compared with the mice of normal contral,comparisons as follows:2.100±0.310 vs 1.536±0.135,0.889±0.183 vs 0.636±0.127,63.070±10.464 vs48.037±6.092;meanwhile respiratory system compliance noteworthily decreased(0.016±0.004 vs 0.023±0.003),and these differences were all statistical significance(P<0.05).(3)Lung tissue homogenate training:the experimental group mice's lung tissue was given homogenate training,and the aspergillus colony was blue-green,while there is no aspergillus colony in normal contral.(4)Histopathology:lung slices in the experimental group mice were watched under the microscope and we could observe the alveolus tissue under the state of congestion and edema,inflammatory cells infiltrating,the alveolar structure broken,emphysema formation,between alveolars space appearing and hyphae divarication presenting 45°angle,all of observations above-mentioned accorded with the pathological features of COPD combined with IPA,but there were no changes in the normal contral.Conclusion:1.COPD combined with IPA usually happen to the old,and compared to the patients suffering COPD alone,they have a higher smoking index and longer hospitalization stay.2.The situations of applying hormones and antibiotics,hospitalizing repeatedly,treating in ICU and malnutrition possess a important portion of the patients of COPD combined with IPA,even affect the occurrence and progress of the diseases in a certain extent.3.The symptoms of COPD combined with IPA are not typical,and specificity andsensitivity of related laboratory inspections and imagining examinations are not high,but repeated sputum culturing has an important significance to diagnosis.4.For the patients diagnosed and suspected,applying standard antifungal treatment in early phase benefits the patients.Meanwhile,antifungal treatment for critical patients has difficulty in comprehensive therapy combining with the patients.5.On the basis of being made to establish a COPD animal model using pure passive smoking method,the mice were given methylprednisolone which induced immunosuppression of the mice,and were inoculated the spore suspension liquid using tracheal puncture method afterwards,which succeeded to establish the mice model of COPD combined with IPA.