| Background and purpose of the research Pneumoconiosis is one of the most common occupational diseases in our country.It is a kind of systemic disease that symptomed mainly as diffuse pulmonary fibrosis,which is caused by the inhalation of dust in the production process and the retention of lung.The most serious harmful to the workers was silicosis,which accounted for more than half of all in the pneumoconiosis cases.Due to the highest prevalence and greatest harmfulness to the workers,the pathogenesis of silicosis has always been the focus of researchers.It has been found that,the inhalation of dust particles in the body can activate the immune cells,which can lead to many autoimmune diseases,including autoimmune diseases.In this study,we will investigate the expression of some autoimmune indexes in silicosis patients: the levels of Rheumatoid factor(RF),anti ds-DNA antibody(anti ds-DNA),transcription factor Foxp3,retinoic acid-related solitary nucleus receptor γt(RORγt),CD4+CD25+ regulatory T lymphocytes(CD4+CD25+regulatory T cells,Tregs),helper T cells 17(T help cell 17,Th17),interleukin 17(IL-17)and the abnormal rate of antinuclear antibody(ANA);And to explore their correlation with the degree of lung lesions.To explore the expression of immune indicators in patients with silicosis and the relationship between the degree of lung lesions in patients with silicosis preliminary.Try to find out objective indexes to help reflect the actual condition of pulmonary lesions in patients with silicosis,and providetheoretical reference for clinical diagnosis,condition judgment,treatment plan improvement and curative effect judgment.Method 1.Object of study One hundred and twenty patients with silicosis were chosen as the case group;Ninety healthy people without occupational hazard factors contract were selected as the health control group.The age,sex,smoking habit,past disease history of the two groups were collected.Collect the type of work,working age,silicosis diagnosis period and the overall intensity in the case group.2.The experimental method 2.1 Blood specimen collection Collecting 5ml/10 ml morning fasting whole blood in the case group patients on the 2nd day after admission and 5ml/10 ml morning fasting whole blood in the health control group respectively,taking serum for inspection after centrifugation.2.2 The expression level of rheumatoid factor was detected by immune turbidimetry,anti ds-DNA antibody was detected by enzyme linked immunosorbent assay,ANA titer was detected by indirect immunofluorescence assay,the expression of transcription factors Foxp3 and RORγt was detected by Real-time PCR,the expression of regulatory T cells and helper T cells 17 were detected by flow cytometry,the level of serum IL-17 cytokines was detected by ELISA.2.3 Blood gas analysis was performed on the second day after admission,partial pressure of carbon dioxide(PCO2)and partial pressure of oxygen(PO2)were collected in the silicosis group.2.4 Pulmonary ventilation function tests were carried on the second day of hospitalization in the case group,the test items included forced vital capacity(FVC),forced expiratory volume(FEV1),a second rate(FEV1%),peak expiratory flow(PEF),peak expiratory flow(FEF25~75%);forced expiratory flow rate at 25% of vital capacity(FEF25%),instantaneous flow rate 50% forced expiratory vital capacity(FEF50%),forced expiratory flow rate at 75% of vital capacity(FEF75%),forced expiratory volumeaccounted for the percentage of vital capacity(FEV1/VC%).3.Statistical methods SPSS 19.0 software was used for statistical analysis.All measurement data were tested by normality(KS test).The normal distribution was expressed by the mean addition and subtraction standard deviation,the data of non-normal distribution were described by M(P25,P75).The comparison between the two groups by using the Wilcoxon rank sum test and t test of two samples,the comparison among multiple groups by using ANOVA or Kruskal Wallis test.The odds ratio was compared by using the Chi-square test or Fisher’s exact probability method.The correlation analysis was performed by using Pearson correlation Analysis or Spearman rank correlation analysis.Result 1.The abnormal rate of ANA in the case group was higher than that in the health control group(P <0.05).There were no significant differences in the abnormal rate of ANA in the different dust-exposing time,silicosis stages,general dispersion in pulmonary area,pulmonary ventilation functions and whether the oxygen partial pressure of blood gas analysis indexes were normal or not(P>0.05).2.The expression levels of RF and anti ds-DNA antibodies in the serum were in normal range both of the case group and the health control group,but there was no significant difference in the expression of RF and anti ds-DNA antibodies between the two groups(P>0.05).There was no significant difference in the levels of RF and anti ds-DNA antibodies among different stages and different total density of silicosis patients(P>0.05).There is a weak positive correlation between the expression of RF and the age in silicosis patients(rs=0.227,P<0.05),but there were no correlations with silica dust –exposing time,the percentage of pulmonary function measured value accounted for predicted value,the partial pressure of carbon dioxide and partial pressure of oxygen(P>0.05).There were no correlations among the expression level of anti ds-DNA antibodies and the age,the time of silica-exposing time,the percentage of pulmonary function measured value accounted for predicted value,the partial pressure of carbon dioxide and partial pressure of oxygen(P>0.05).3.The expression level of Foxp3 and RORγt in serum of the patients was higher than that of the health control group(P<0.05).The expression level of Foxp3 in the third phase of silicosis patients was higher than that of the stage one and stage two(P< 0.05).The expression level of RORγt in the third phase of silicosis patients was higher than that of the stage one(P<0.05).There was no significant difference in expression level of Foxp3 and RORγt between the silicosis patients of stage one and stage two(P>0.05).There was no significant difference in expression level of RORγt between the silicosis patients of stage two and stage three(P>0.05).There was no significant difference in expression level of Foxp3 and RORγt among silicosis patients with different general dispersion in pulmonary area(P>0.05).The age of patients,dust-exposing time,the pulmonary function,the blood gas analysis had no correlations with the expression levels of Foxp3 and RORγt in silicosis patients(P>0.05).4.The expression level of Treg and Th17 in serum of the patients was higher than that of the health control group(P<0.05).There was no significant difference in expression level of Treg and Th17 between the silicosis patients of stage one and stage two plus stage three(P>0.05).There was a negative correlation between the expression level of Treg and silica dust-exposing time in silicosis patients(rs=0.390,P<0.05);There was a positive correlation between the expression level of Treg and the indexes such as forced vital capacity,forced expiratory volume of pulmonary ventilation function in silicosis patients(rs=0.487or0.460 respectively,P<0.05);There were no correlation among the expression level of Treg with the age,the partial pressure of carbon dioxide,the partial pressure of oxygen and other indexes of pulmonary ventilation function in silicosis patients(P>0.05).There were no correlation among the expression level of Th17 with the age,the silica dust-exposing time,the pulmonary ventilation function and the blood gas analysis in silicosis patients(P>0.05).5.The expression levels of IL-17 in the serum of the case group were significantly higher than that in the health control group(P<0.05);There was no significant difference in expression level of IL-17 between different silicosis stages and general dispersion in pulmonary area(P>0.05).There was no correlation among the expression level of IL-17with the age,the silica dust-exposing time,the pulmonary ventilation function and the blood gas analysis(P>0.05).Conclusion 1.There were immune disorders and inflammations existence in some silicosis patients,which could induced autoantibodies.2.The abnormal rate of ANA in serum of silicosis patients was higher than that of normal people,but could not found the correlation with the degree of lung lesions in silicosis patients.3.The expression levels of RF and anti ds-DNA antibodies in the serum of silicosis patients were in normal range of the case group,the level of anti ds-DNA antibody increased with the promoting of silicosis stage,but could not found the correlation with the degree of lesions in silicosis patients.4.The expression levels of Foxp3,RORγt,Treg,Th17 and IL-17 of silicosis patients were higher than those of normal people,the expression levels of Foxp3,RORγt and IL-17 showed an increasing trend with the promoting of silicosis stage;These results indicate that Treg,Th17 and their key transcription factors(Foxp3,RORγt)and cytokine(IL-17)are involved in the immune disturbance in silicosis patients,which that may be related to the severity of silicosis.5.The current study found that,there were some correlations among Treg,Foxp3,RORγt and the degree of lung lesions in silicosis patients,which could be used as the indicators for efficacy judgment. |
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