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1.
Journal of Southern Medical University ; (12): 1530-1545, 2015.
Article in Chinese | WPRIM | ID: wpr-232576

ABSTRACT

<p><b>OBJECTIVE</b>To analyze the quantity and size distribution of 24-hour urinary extracellular vesicles (uEVs) from healthy adults.</p><p><b>METHODS</b>The 24-hour uEVs from 9 healthy adults were isolated by hydrostatic filtration dialysis (HFD). The effectiveness of uEVs enrichment was evaluated using Western blotting and transmission electron microscopy (TEM). The quantity and size distribution of the uEVs was analyzed with BCA protein quantification, TEM, and nanoparticle tracking analysis (NTA).</p><p><b>RESULTS</b>uEVs with different sizes and morphologies were observed under TEM. Western blotting confirmed the expression of TSG101 in all the uEV fractions from the 9 donors, ranging from 132.50 to 760.70 ng/mL. NTA results showed that the number of 24-hour uEVs amount ranged from 3.56 × 10¹² particles to 5.12 × 10¹² particles, with a CV of 14.23%. The proportion of the vesicles with a diameter <40 nm was 0.04%-0.69% with a number range of (1.80-26.49)× 10⁹ particles; the proportion of vesicles with a diameter of 40-100 nm (which is consistent with the size of exosomes)was 22.07%-42.08% with a number range of (1.00-1.77)× 10¹² particles. The proportion of vesicles with a diameter of 100-1000 nm (consistent with the size of microvesicles) was 57.88%-77.85% with a number range of (2.09-3.86)× 10¹² particles.</p><p><b>CONCLUSION</b>The established HFD method allows efficient and convenient isolation of uEVs from a large amount of urine samples. The 24-hour uEVs from healthy adults show narrow differences between individuals and thus can be an ideal source of samples for relevant studies.</p>


Subject(s)
Adult , Humans , Blotting, Western , Cell-Derived Microparticles , Exosomes , Extracellular Vesicles , Microscopy, Electron, Transmission , Nanoparticles , Urine
2.
Journal of Southern Medical University ; (12): 1683-1688, 2015.
Article in Chinese | WPRIM | ID: wpr-232546

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the value of serum IgA/C3 ratio in the diagnosis of IgA nephropathy and explore its relationship with the clinicopathological features of the patients.</p><p><b>METHODS</b>Sixty-six patients with IgA nephropathy, 111 with other glomerular diseases, and 40 healthy control subjects without kidney disease were tested for serum IgA and C3 levels using CRM470 adjusted standardized immune turbidimetric method, and the IgA/C3 ratio was calculated. According to Oxford and Lee's classification criteria, we analyzed the pathological grades of the renal biopsy samples from patients with IgA nephropathy. The ROC curve was used to assess the value of serum IgA and IgA/C3 ratio in predicting IgA nephropathy.</p><p><b>RESULTS</b>Patients with IgA nephropathy had an elevated serum IgA/C3 ratio than those with other glomerular diseases and the control subjects, with an area under the ROC curve of 0.776. An elevated serum IgA/C3 ratio was not found to significantly correlate with the pathological grade of renal biopsy samples in patients with IgA nephropathy.</p><p><b>CONCLUSION</b>In the absence of renal biopsy findings, serum IgA/C3 ratio can help in the diagnosis of IgA nephropathy.</p>


Subject(s)
Humans , Biopsy , Case-Control Studies , Complement C3 , Glomerulonephritis, IGA , Blood , Diagnosis , Immunoglobulin A , Blood , Kidney , Pathology
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