| Part ?:hsa-mir-7 is a potential biomarker for human idiopathic inflammatory myopathy with interstitial lung diseasePurpose:Idiopathic inflammatory myopathies(IIMs)are a rare group of autoimmune diseases characterized by skeletal muscle weakness and inflammation.IIMs mainly include polymyositis(PM),immune-mediated necrotizing myopathy(IMNM),sporadic inclusion-body myositis(sIBM),non-specific muscles.Nonspeific myositis(NSM)and dermatomyositis(DM).MicroRNAs(miRNAs)regulate a wide range of developmental and physiological cellular processes.Research on new methods for the generation and diagnosis of IIMs,such as the study of miRNAs,is critical for the development of new treatments for IIMS and/or better treatment and diagnosis.The significance of circulating miRNAs in the diagnosis of idiopathic inflammatory myopathy(IIMs)is unclear.In this study,we aimed to investigate the significance of circulating miRNAs as potential biomarkers for predicting IMMs and interstitial lung disease(ILD)patients.Methods:The experiment was divided into two groups:study group and control group.The patients were admitted to our hospital from March 2015 to February 2018,and were diagnosed as idiopathic inflammatory myopathy by clinical and pathological diagnosis.A total of 43 patients with newly diagnosed IIMs without any treatment were enrolled.Of these,13 patients with IMMs were diagnosed with PM and the remaining 30 were diagnosed with DM.Both PM and DM patients met Bohan&Peter or Dalakas diagnostic criteria.In addition,serum samples from 43 healthy people were collected as controls.All plasma samples collected in this study were informed consent of the participants and reviewed by the hospital ethics committee.Total RNA was then extracted from serum samples from 43 IIMs patients and 43 healthy individuals,respectively.miRNA microarray chips were used to screen and analyze the differences in miRNAs expression profiles in serum samples from both groups,and finally verified by qRT-PCR.Results:Microarray showed that the expression profile of circulating miRNAs in patients with IIM was significantly different from that in healthy controls(P<0.05).qRT-PCR confirmed a significant difference in mir-7 and mir-21 levels in plasma samples from patients with IIM and healthy controls(p<0.05).However,only miR-7 was over-expressed in every IIM patient(P<0.05),and miR-7 was lower in the serum of IIM/ILD patients than in patients without ILD(P<0.05).The area under the curve(AUC)for IIM/ILD patients and IIMs patients without ILD was 0.8978,with a 95%confidence interval of 0.7961 to 0.9995.The receiver operating characteristic curve(ROC)analysis indicated that the cutoff value of miR-7 was 0.0063.Furthermore,AUC analysis showed that both miR-7 and miR-21 have diagnostic value for patients with IMMs from health control;however,miR-7 is more sensitive.Conclusion:Circulating miR-7 is a potential biological diagnostic marker for patients with IIM and can be used to distinguish between IIM/ILD patients and IIM patients who do not have ILD.Part ?:B cell activating factor(BAFF)and the progression of idiopathic inflammatory myopathyObjective:Idiopathic inflammatory myopathy is a histopathological diagnosis that analyzes changes in molecular immunopathology in patients with idiopathic inflammatory myopathy,contributes to disease diagnosis,and provides more reliable clinical treatment.according to.This experiment will evaluate the importance of plasma B cell activating factor(BAFF)expression levels in the progression of idiopathic inflammatory myopathy.Methods:Select our hospital from 2015 to 2018 A total of 61 patients with IIMs admitted to the hospital were treated with prednisone alone orally,except for inclusion body myositis(IBM).The specific oral doses are as follows.The initial dose of prednisone was 1 mg/kg morning,and the degree of improvement of muscle strength and the change of plasma CK expression level were observed.When the muscle strength was improved and the plasma CK expression decreased,the oral dose of prednisone was reduced by 5 mg/w.Until the oral dose is maintained at 20-30 mg/qod.Serum BAFF level was detected by ELISA,and the expression of related molecules in muscle tissue was detected by immunohistochemistry.Results:Of the 61 patients with IIMs,21 patients with IIMs were diagnosed with PM,and the remaining 40 patients were diagnosed with DM.The expression level of plasma B cell activating factor(BAFF)in patients with dermatomyositis(DM)before treatment was(3673±778)pg/ml,and the expression level of plasma BAFF in patients with polymyositis(PM)was(1673±621).)pg/ml,while the serum BAFF expression level in the normal control group was only(681±107)pg/ml.Statistical analysis showed that the serum BAFF expression level in patients with DM or PM was significantly higher than that in normal healthy patients(P<0.05).At the same time,the serum BAFF expression level in DM patients was significantly higher than that in PM patients(P<0.05).After treatment with strong pines,the serum BAFF expression level was(763±106)pg/ml in the DM group and the serum BAFF expression level in the PM group was(661±87)pg/ml.Statistical analysis showed that the expression of BAFF in serum of patients with DM or PM decreased significantly before and after treatment with strong pine(P<0.05).Conclusion:BAFF,a member of the tumor necrosis factor superfamily,is involved in the progression of DM and PM in patients with idiopathic inflammatory myopathy.The expression level is negatively correlated with disease outcome.By detecting the expression level of plasma BAFF,it can be clinical.To provide an accurate reference for judging the therapeutic effects of DM and PM patients.Part ?:Exploring the application value of cytokines and costimulatory molecules in the diagnosis and differential diagnosis of idiopathic inflammatory myopathyObjective:To investigate the application value of cytokines CD4 and CD8 in muscle tissue and peripheral blood co-stimulatory molecules CD4-ICOS,CD4-OX40,CD8-ICOS,CD8-OX40 and CD19-CD40 in the diagnosis and differential diagnosis of IIMs.Methods:Thirty-six patients with IIMs admitted to the First Affiliated Hospital of Suzhou University from march 2015 to february 2018,were confirmed by muscle biopsy,including 26 patients with PM and 10 patients with DM.They were 43.7 ± 6.9 years old and 46.5 ± 7.2 years old.At the same time,36 patients with other systemic diseases with normal muscle pathology but obvious muscle weakness were used as negative control group.The muscle tissue samples of the above subjects were preserved by frozen section embedding,and the blood samples were separated and serum was stored at-196 degrees liquid nitrogen.The clinical,histological and immunopathological features of each subgroup of IIMs were identified.Immunohistochemical staining(IHC)was used to detect the expression levels of CD4 and CD8 cytokines in idiopathic inflammatory myopathy and control populations.Flow cytometry was used to detect the expression levels of peripheral blood co-stimulatory molecules CD4-ICOS,CD4-OX40,CD8-ICOS,CD8-OX40,CD19-CD40 in various groups of idiopathic inflammatory myopathy and control population.Results:Immunohistochemical staining of muscle tissue in each subgroup of IIMs suggested that there was significant inflammatory cell infiltration in the muscle tissue of IMMs,and the distribution of CD4 and CD8 in muscle tissue of DM and PM patients was completely different.Inflammatory cell infiltration of CD8+T is the main,while DM muscle tissue is mainly infiltrated by inflammatory cells of CD4+T.At the same time,the experiment showed that no significant inflammatory cell infiltration was observed in the negative control group.Peripheral blood flow cytometry detection of pre-treatment co-stimulatory molecules suggests that there is also a significant difference in the expression of costimulatory molecules between IIMs patients and negative control groups,compared with the negative control group,CD4-ICOS,CD4-OX40,CD8-ICOS,CD8-OX40 was significantly elevated in the serum of PM patients,while CD19-CD40 was significantly elevated in the serum of DM patients.Further analysis of subgroups of patients with IIMs suggested that CD4-ICOS,CD4-OX40,CD8-ICOS,and CD8-OX40 were significantly higher in peripheral blood of PM patients than in DM patients,while CD19-CD40 was significantly expressed in peripheral blood of DM patients.Higher than PM patients.Flow cytometry analysis before and after treatment of patients with PM and DM suggested that the cost of co-stimulatory molecules including CD4-ICOS,CD4-OX40,CD8-ICOS,CD8-OX40 was significantly decreased in the serum of patients after treatment,and at the same time,in DM The co-stimulatory molecule CD 19-CD40,which is significantly elevated in the serum of patients,also showed a significant decrease.Conclusions:1.The proportion of CD4+T/CD8+T cells is different in muscle tissue of DM and PM patients.DM is mainly CD4+T cells,and PM is mainly CD8+T cells.Thus,the ratio of CD4+T/CD8+ T cells can be used to identify DM and PM subgroups in patients with IIMs.2.The differential expression of peripheral blood co-stimulatory molecules in IIMs patients and negative control group suggests that costimulatory molecules participate in the process of IIMs patients.The peripheral blood of PM patients is mainly CD4-ICOS,CD4-OX40,CD8-ICOS,CD8-OX40.The expression of CD19-CD40 in peripheral blood of patients with DM was elevated.Thus,peripheral blood co-stimulatory molecule expression classes can also be used to identify DM and PM subgroups in IIMs patients.The significant decrease in costimulatory molecules after hormone therapy also suggests that the expression level of costimulatory molecules in IMMs can also indirectly reflect the improvement of hormone therapy in patients with IIMs. |
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