Serum exosomal microRNAs as potent circulating biomarkers for melanoma.

Exosomes are small homogenous membrane vesicles that derive from the exocytosis process of cells and can contain DNA, microRNAs (miRNAs), and/or proteins. Characterization of the content profile of exosomes may reflect the state of the cells that release them, and this could be predictive of disease. In this study, to explore the potential biomarkers for melanoma, we isolated serous exosomes from 30 patients with melanoma and 30 healthy individuals using the ultracentrifugation method. Five miRNAs were subsequently detected in each sample by quantitative reverse transcription-PCR: miRNA-532-5p, miRNA-106b, miRNA-200c, miRNA-199a-5p, and miRNA-210. Only the levels of exo-miRNA-532-5p and exo-miRNA-106b differed between the two groups (Z=-4.17 and -4.57, respectively, P<0.0001). When these two miRNAs were evaluated individually and in combination in 95 melanoma patients and 95 healthy individuals serum samples, the area under the receiver operating characteristic curve values were 0.867, 0.820, and 0.936, respectively. Furthermore, in blinded tests of samples from 25 melanoma patients and 25 healthy individuals, this panel of miRNAs identified 23/25 patients with melanoma (92.0% sensitivity) and 22/25 healthy individuals (88.0% sensitivity). Our exo-miRNA panel also distinguished patients with metastasis from those without metastasis, patients with stage I-II disease from those with stage III-IV disease, and patients who had received pembrolizumab treatment from those who were untreated. Overall, these results indicate that serum exosomal miRNAs, especially exo-miRNA-532-5p and exo-miRNA-106b, have the potential to be used for monitoring and/or a diagnosis of melanoma in a clinical setting.

Increasing Expression of IL-3 7 in Peripheral Blood Mononuclear Cell of Patients with Primary Biliary Cirrhosis and Its Clinical Significance / 现代检验医学杂志

Objective To test the expression level of IL-37 in peripheral blood mononuclear cells (PBMCs)of patients with primary biliary cirrhosis (PBC)and further explore its clinical significance in the pathological process of PBC.Methods Pe-ripheral blood samples were collected from 42 patients diagnosed as PBC and 38 health individuals examined at the same time during June 2013 to August 2015 in Changhai Hospital.PBMCs were separated by sucrose density gradient centrifugation, qualified Real Time-Polymerase Chain Reaction (qRT-PCR)was used to measure IL-37 mRNA expression level in PBMCs. Enzyme-Linked Immuno Sorbent Assay (ELISA)was to measure the protein level of IL-37,IL-6,IL-17,TNF-α,TGF-β,IL-18 and IL-23 in plasma.Meanwhile,the pathological stages of PBC cases were recorded.Pearson correlation analysis was performed on IL-37 and IL-6,TNF-α,IL-17,TGF-β,IL-18 and IL-23.Spearman rank correlation analysis was on IL-37 and pathological stages of PBC.Results The mRNA and protein level of IL-37 in experimental and controlled group were 2.81 ±0.94 vs 1.09±0.56,356.14±169.36 pg/ml vs 86.68±48.23 pg/ml separately(t=9.811,9.462,P<0.000 1),with sta-tistical differences.The correlation analysis showed that IL-37 was positively related with IL-17,TNF-α,IL-6 and TGF-β(r=0.561 2,0.661 9,0.672 1,0.765 3,P<0.001),and disease stages (Ⅰ~Ⅳ)(rs=0.348 9,P<0.05).Conclusion IL-37 might involve in the pathological process of PBC,and it is significant for disease prediction and diagnosis.

Elevated Expression of CD69 +CD4+CD25-T Cells in peripheral Blood from Patients with Autoimmune Pancreatitis and Its Clinical Significance / 现代检验医学杂志

Objective To test the expression level of CD69+CD4+CD25-T cells in peripheral blood from patients with autoimmune pancreatitis,and further analyze its clinical significance.Methods Peripheral blood samples from 32 patients with AIP diagnosed in hematological department,Changhai Hospital and 32 health individuals examined at the same time were collected from September 2014 to December 2016,they were classified as experimental and control groups,separately.Peripheral blood mononuclear cells (PBMCs) was acquired by density gradient centrifugation,CD69+ CD4 + CD25-T cells in PBMCs were tested by flow cytometry,and the expression level of cytokines in plasm was by ELISA.The comparison of varies between the two groups was measured by two independent samples' t test.The relationship between the two measurement data was measured by pearson correlation coefficient.Results The expression levels of CD69 + CD4 + CD25-T in experimental and control groups were 10.36％±3.68％ vs 3.99％±1.45％ (t=9.110,P＜0.0001).The expression level of TGF-β was 399.86±121.88 vs 143.87±56.22 pg/ml (t=10.79,P＜0.000 1),both with statistical significance.The levels of CD69+CD4+CD25-T in experimental was positively correlated with TGF-β (r=0.653,P＜0.001) and negatively with IL-4,IFN-γ,IL-2 (r=-0.442,-0.567,-0.351,P＜0.05) and there was statistical significance.Conclusion CD69+CD4 +CD25-T cells might involve the immunopathology of AIP and could be the potential biomarker for clinical diagnosis and therapy.

Affection of CD2 6/DPP4 on CD4+T Cells and Relative Cytokines in Patients with Crytococcal Meningitis and Its Clinical Significance / 现代检验医学杂志

Objective To analyze the affection and clinical significance of CD26/DPP4 on CD4+T cells and its cytokines in patients withCrytococcalMeningitis.Methods Peripheral blood was collected from 36 patients diagnosed withCrytococcal Meningitis in Changhai Hospital and Changzheng Hospital,Shanghai from August,2011 to December,2015,meanwhile 36 health controls’was also acquired.Peripheral blood mononuclear cell (PBMC)was separated by density gradient centrifuga-tion,CD26+CD4+T and CD26-CD4+T cell groups were classified by Flow Cytometry,the expression level of cytokines was tested by reverse transcriptase-polymerase chain reaction (RT-PCR).The correlation between DPP4 activity,CD26+CD4+T (%)and APACHE II score,IL-17,TNF-α,IL-4,IFN-γwas measured by Pearson coefficient.Results CD26+CD4+T(%)between experimental and control groups was 13.35±3.83 vs 8.39±2.14 (t=6.78,P<0.000 1).DPP4 activity was 50.89±17.21 mU/ml vs 73.83±20.24 mU/ml (t=5.18,P<0.000 1),with statistically significant differences.In ex-perimental groups,CD26+CD4+T (%)was positively related with APACHE II score,IL-17,TNF-α(r=0.431,0.564, 0.688,P=0.003 8,0.001,0.004 6).DPP4 activity was negatively interrelated with APACHE II score,IL-17,TNF-α,IFN-γ(r=-0.544,-0.489,-0.678,-0.734;P<0.001).Conclusion CD26/DPP4 may be involved in the pathogenesis of Crytococcal Meningitis through regulation of Th subgroups,and it was the potential therapeutic target and the predicted marker of the disease.