Clinical Study Of Pathogen Distribution,T Cell Subsets And Cytokine Levels And Their Intervention In Patients With COPD

Part?Distribution characteristics of pathogens in COPD patientsObjective:To understand the distribution of common pathogenic bacteria,atypical pathogens and respiratory viruses in patients with chronic obstructive pulmonary disease(COPD)in different periods of this hospital,to analyze the infection status and changes of COPD patients in different periods,and to explore the relationship between pathogens and the progression of COPD,so as to provide a theoretical basis for clinical rational drug use.Methods:We prospectively selected and recruited 150 stable COPD patients and 150patients with acute exacerbation of chronic obstructive pulmonary disease(AECOPD)respectively,who came from outpatient,emergency or inpatient department in the People's Hospital Affiliated to Nanchang University from March 2018 to December2019,and collected the general clinical data of the patients.Collect sputum samples from 2 groups of patients for bacteriological culture and strain identification;collect peripheral venous blood of 2 groups of patients,and detect IgM antibodies of 8common pathogens of respiratory tract infection in serum by indirect immunofluorescence test(IFA),including atypical pathogens:Legionella pneumoniae(LP),Mycoplasma pneumoniae(MP),Chlamydia pneumoniae(CP);and viral antibodies:influenza virus A(F1u A),influenza virus B(F1u B),parainfluenza virus(PIV),respiratory syncytial virus(RSV),adenovirus(ADV).Analyze and compare the distribution characteristics and differences of pathogens between groups.Results:1 General data of the research subjects:a total of 300 patients with COPD were included,including 230 males(76.7%),age M(IQR)was 68.0(61.0,75.0)years,and course M(IQR)was 12.0(7.0,17.0)years,There was no statistical difference in the general clinical characteristics of the 2 groups of patients(p>0.05).2.Results of sputum culture:distribution of pathogenic bacteria in stable COPD group:57 cases were positive for sputum culture,the detection rate was 38.0%,73pathogenic bacteria were detected,including 41 single pathogens,accounting for71.9%,and 16 double pathogens,accounting for 28.1%.47 strains of Gram-negative bacilli were detected,accounting for 64.4%.The top three common strains were Haemophilus influenzae,Klebsiella pneumoniae,Pseudomonas aeruginosa;25 strains of Gram-positive bacteria,accounting for 34.2%,The top three common strains are Staphylococcus aureus,Streptococcus pneumoniae,and Streptococcus hemolyticus;one strain of fungus is Candida albicans,accounting for 1.4%.The distribution of pathogenic bacteria in the AECOPD group:101 cases were positive for sputum culture,the detection rate was 67.3%,and 129 pathogenic bacteria were detected,of which 73were single pathogens,accounting for 72.3%,and 28 were double pathogens,accounting for 27.7%.98 strains of Gram-negative bacilli were detected,accounting for 76.0%.The top three common strains were Klebsiella pneumoniae,Haemophilus influenzae,and Pseudomonas aeruginosa;27 strains of Gram-positive bacteria,accounting for 20.9%,the top three common strains are S.pneumoniae,S.aureus,and hemolytic streptococcus;4 strains of fungi,accounting for 3.1%,are 3 Candida albicans and 1 Aspergillus.The positive rate of sputum culture in the AECOPD group(67.3%vs 38.0%,?~2=25.887,p<0.05)was significantly higher than that in the stable COPD group.3 Results of atypical pathogens:25 patients with positive tests in the stable COPD group,with a positive rate of 17.7%,followed by 14 MPs,and 11 LP;49 patients with positive tests in the AECOPD group,with a positive rate of 32.7%,followed by 29cases of MP,1 case of CP,and 19 cases of LP.Counting the infection of pathogenic microorganisms in the 2 groups,it was found that the total positive rate of the AECOPD group,MP and LP positive rates were significantly higher than the stable COPD group,p<0.05.4 Common respiratory virus results:12 patients with positive test in stable COPD group,the positive rate was 8%,followed by Flu A 2 cases,Flu B 4 cases,PIV 2 cases,RSV 1 case,ADV 3 cases;AECOPD group patients 33 cases were tested positive,the positive rate was 22%,followed by 9 cases of Flu A,18 cases of Flu B,3 cases of PIV,2 cases of RSV,and 1 case of ADV.The total positive rate,Flu A and Flu B positive rate of AECOPD group were significantly higher than those of stable group,p<0.05.Conclusion:1 Pathogens can be detected in patients with COPD at different times in this area,and some colonization of lower respiratory tract in stable COPD patients.The detection rate of patients in the AECOPD group was higher than that in the stable COPD group.Common pathogens were Gram-negative bacteria.Clinical treatment should be guided by the results of pathogen culture and antibiotics should be selected rationally.2 Among COPD patients in this area,the infection rate of atypical pathogens in the AECOPD group was significantly higher than that in the stable COPD group.MP and LP were more common,suggesting that atypical pathogens participate in the acute exacerbation mechanism of COPD.3 The total virus detection rate of the AECOPD group in this area is significantly higher than that in the stable phase group.Viral infection is closely related to the acute exacerbation of COPD.AECOPD major group virus Flu A,Flu B,PIV;stable COPD major group virus Flu B,ADV,Flu A.Part II Study on T lymphocyte subsets and cytokine levels in COPD patientsObjective:By detecting the characteristics of peripheral blood T-lymphocyte subsets and the levels of IL-4,IL-6,IL-8,IL-17 in COPD patients,the differences among the above indexes were compared,and the clinical significance of these indexes in COPD patients at different stages was discussed to guide the immune prevention and clinical treatment of COPD.Methods: We prospectively selected and recruited 150 stable COPD patients and 150 patients with acute exacerbation of chronic obstructive pulmonary disease(AECOPD)respectively,who came from outpatient,emergency or inpatient department in the People's Hospital Affiliated to Nanchang University from March 2018 to December 2019,and 150 healthy people who had physical examinations in our hospital during the same period were selected as healthy controls.Collect general clinical data of the research subjects,including gender,age,body mass index,course of disease,smoking history,etc.The peripheral venous blood of three groups of subjects was collected,and the expression of CD3+,CD4+,CD8+ in peripheral blood T lymphocyte subsets was detected by flow cytometry,and the ratio of CD4+ / CD8+ was calculated;the cytokines IL-4,IL-6,IL-8,IL-17 were measured by enzyme-linked immunosorbent assay(ELISA),and the differences of the observation indexes between the three groups were compared and analyzed.Results: 1 General data of the research subjects: a total of 300 patients with COPD were included,including 230 males(76.7%),M(IQR)age was 68.0(61.0,75.0)years,and M(IQR)course was 12.0(7.0,17.0)years.Of the 150 healthy controls,105 were male(70.0%),the age M(IQR)was 65.0(60.0,70.0)years old,and the BMI was(23.6 ± 3.7)kg/m2.There were no significant differences in gender,age and BMI between the three groups(p > 0.05).2 Results of T lymphocyte subsets in peripheral blood: compared with the levels of CD3+(%),CD4+(%),CD4+ / CD8+(47.03 ± 9.11,29.34 ± 8.86,0.98 ± 0.29)in the AECOPD group,the levels in the stable COPD group(53.83±10.48,35.65±6.59,1.40±0.27)and the healthy control group(66.17±8.59,42.41±8.12,2.02±0.45)were significantly higher than those in the AECOPD group(p < 0.05);compared with the AECOPD group CD8+(%)(29.87 ± 6.37),the stable COPD patients(25.42 ± 6.91),the healthy control group(20.95 ± 6.37)were decreased significantly(p <0.05).Compared with the levels of CD3 +(%),CD4 +(%),CD4 + / CD8+(53.83 ± 10.48,35.65 ± 6.59,1.40 ± 0.27)in the stable COPD group,the level of healthy control group(66.17 ± 8.59,42.41 ± 8.12,2.02 ± 0.45)significantly increased(p <0.05);compared with CD8 +(%)(25.42 ± 6.91)in stable COPD patients,the healthy control group(20.95 ± 6.37)decreased significantly(p <0.05).3 Results of peripheral blood cytokines: The levels of IL-4,IL-6,IL-8 and IL-17 in AECOPD group(92.93 ± 23.46,73.81 ± 16.68,92.34 ± 16.65,77.25 ± 22.38)(PG / ml)were higher than those in stable COPD group(71.25 ± 20.13,35.87 ± 5.74,47.54 ± 6.91,59.61 ± 12.57)(pg/ml)and healthy control group(48.73 ± 8.96,9.67 ± 1.89,25.63 ± 5.53,23.76 ± 6.54).The levels of IL-4,IL-6,IL-8 and IL-17 in the stable COPD group(71.25 ± 20.13,35.87 ± 5.74,47.54 ± 6.91,59.61 ± 12.57)(pg / ml)were significantly higher than those in the healthy control group(48.73 ± 8.96,9.67 ± 1.89,25.63 ± 5.53,23.76 ± 6.54)(pg / ml),and the difference was statistically significant(p< 0.05).Conclusion: 1 Detection of T lymphocytes and cytokines in peripheral blood of patients with COPD helps to further study the role of immune function in the pathogenesis.2 COPD patients have abnormal cellular immune function,which is characterized by dysfunction of T lymphocytes and disorder of inflammatory cytokines.3 Compared with stable COPD patients,T-lymphocyte dysfunction and inflammatory cytokine disorder are more significant in AECOPD patients,suggesting that the abnormal cellular immune function plays an important role in the pathophysiological process of chronic airway inflammation,and its level change can be used as a monitoring index to judge the condition and prognosis.Part III The effects of budesonide/formoterol and thymosin on immune function in patients with COPDObjective: To explore the effects of budesonide/formoterol(BUD / FM)and thymosin on the peripheral blood T lymphocyte subsets and cytokines in patients with stable COPD.Methods:80 patients of stable COPD of outpatient,emergency and inpatient department in People's Hospital affiliated to Nanchang University from March 2018 to December 2019 were screened.The acquired factors such as gender,age,body mass index,course of disease,smoking history and complications were collected.A prospective,randomized,parallel,open and controlled study was adopted.A random coding table was prepared by specially assigned persons(those who did not participate in the trial).They were randomly divided into BUD/FM group and thymosin group according to the ratio of 1:1,40 cases in each group.On the basis of routine treatment,the patients in the BUD/FM group were given the powder inhaler of BUD/FM(320ug: 9ug * 60 inhalation * 1 bottle/box),1 inhalation /time,twice a day,for 12 weeks;the patients in the thymosin group were given the subcutaneous injection of thymosin ? 1 at a dose of 1.6mg/time,twice a week,for 12 weeks.And regular clinical follow-up.Before and after the intervention,the levels of CD3+,CD4+,CD8+ and the ratio of CD4+ /CD8+ were measured,the levels of IL-4,IL-6,IL-8 and IL-17 were measured,the physical examination,vital signs,blood routine,urine routine,liver and kidney functions and adverse reactions were observed.Results: 1 general data: A total of 80 patients with stable COPD were included,including 57 males(71.3%),age M(IQR)was 68.0(52.0,74.0)years old,BMI was(24.4 ± 5.3)kg / m2,course M(IQR)is 12.0(7.0,17.0)years.There was no statistical difference in the comparison of general data between the two groups of patients(p> 0.05),which was comparable.The patients in both groups completed the whole treatment process,and no cases of shedding occurred.2 Comparison of T lymphocyte levels and changes before and after treatment in the 2 groups: There was no significant difference(p> 0.05)in the T lymphocyte subsets between the 2 groups before treatment,and they were comparable.After 12 weeks of intervention,the CD3+(%),CD4+(%),CD4+/CD8+ of the BUD / FM group were higher than before treatment,and the changes were 7.86 ± 2.57,4.54 ± 1.59,0.33 ± 0.12 respectively,and CD8+(%)decreased compared with those before treatment,the change values were 4.39 ± 1.43,with statistical significance(p <0.05).CD3+(%),CD4+(%),CD4+/CD8+ of thymosin group all increased from before treatment,the change values were 3.65 ± 1.23,2.18 ± 0.81,0.15 ± 0.06,CD8 +(%)decreased from before treatment,the change value was 1.97 ± 0.72,the differences were statistically significant(p <0.05).After the treatment,the change values of BUD / FM group were significantly higher than that of thymosin group,the differences were statistically significant(p <0.05).3 Comparison of cytokine levels and changes before and after treatment in the two groups: There was no significant difference in cytokine levels between the two groups before treatment(p> 0.05),which was comparable.After 12 weeks of intervention,IL-4,IL-6,IL-8 and IL-17 in the BUD/FM group decreased compared with before treatment,with changes of 18.28 ± 4.35,16.19 ± 5.76,17.93 ± 5.81,21.64 ± 4.80,respectively.(?±s,pg/ml),the differences were statistically significant(p <0.05).IL-4,IL-6,IL-8 and IL-17 in thymosin group decreased compared with before treatment,and the changes were 9.54 ± 3.36,7.87 ± 2.61,7.80 ± 2.79,15.39 ± 4.53(?±s,pg/ml),the differences are statistically significant(p <0.05).After the treatment,the change value of BUD/FM group was significantly higher than that of thymosin group,the difference was statistically significant(p <0.05).4 There were no obvious abnormal changes in blood routine,urine routine,liver and kidney function,blood glucose and blood lipids and other biochemical indicators,chest radiographs,and electrocardiogram before and after treatment in the 2 groups.The patients in both groups did not have serious side effects during treatment.Conclusion: 1 In patients with stable COPD,BUD/FM inhalation or thymosin ?1 continuous intervention for 12 weeks can improve the imbalance of peripheral blood T lymphocyte subsets,correct serum cytokine imbalance,inhibit inflammation,and do not increase the incidence of adverse drug reactions.2 In patients with stable COPD,BUD/FM inhalation improves T lymphocyte subset imbalance and corrects cytokine disorders more effectively than thymosin ?1.