OBJECTIVE： To investigate the effects of tacrolimus combined with valsartan in immunosuppressive regimen on renal function， lipid metabolism and matrix metalloproteinase 2 （MMP-2）， MMP-9， tissue inhibitors of matrix metalloproteinases 2 （TIMP-2） and transforming growth factor-β （TGF-β） in patients with chronic allograft dysfunction （CAD）. METHODS： CAD patients admitted to nephrology department of our hospital from Mar. 2016 to Jun. 2018 were enrolled in group A， B， and C according to the random number table， 34 cases in each group. Group A was given cyclosporin A+Mycophenolate capsules+Prednisone acetate tablets； group B was treated with tacrolimus+Mycophenolate capsules+Prednisone acetate tablets； group C was treated with valsartan on the basis of group B； they were treated for continuous 3 months. Renal function indexes （24 h urinary protein， Scr）， lipid metabolism indicators （TC， TG， HDL， LDL） and the levels of MMP-2， MMP-9， TIMP-2 and TGF-β were compared among those groups. RESULTS： Before treatment， there was no statistical significance in above indexes among 3 groups （P＞0.05）. After treatment， 24 h urinary protein， Scr and TC levels of group A were still higher than normal level， while other lipid metabolism indexes were within normal range. 24 h urinary protein and TC level of group B were still higher than normal level， while Scr level was near the upper limit of the normal range， and other lipid metabolism indicators were within the normal range. Renal function indexes and lipid metabolism indexes of group C were within normal range， while renal function indexes levels of it were lower than those of group B （P＜0.05）； there was no statistical significance in lipid metabolism indexes， compared with group B （P＞0.05）. Compared with before treatment， TGF-β level of group A was significantly increased （P＜0.05）； there was no statistical significance in MMP-2， MMP-9 or TIMP-2 levels （P＞0.05）. TGF-β level of group B and group C was increased significantly （P＜0.05）， while the levels of MMP-2， MMP-9 and TIMP-2 were decreased significantly （P＜0.05）. CONCLUSIONS： Tacrolimus combined with valsartan can effectively delay renal dysfunction and improve lipid metabolism in CAD patients. The mechanism may be related to the inhibition of TIMP-2， MMP-2， MMP-9 and TGF-β expression.

OBJECTIVETo explore the value of detecting podocalyxin (PCX) level in urinary extracellular vesicles for the diagnosis of diabetic nephropathy.

METHODSThis study was conducted among 57 diabetic patients admitted during the period from March to September, 2017, including 34 with uncomplicated diabetics and 23 with diabetic nephropathy; 21 patients with other types of nephropathy and 11 healthy individuals were also included to serve as the controls. Transmission electron microscopy (TEM) and nanoparticle tracking analysis (NTA) were used to verify the separation of urinary extracellular vesicles. The molecular markers of extracellular vesicles (TSG101 and podocalyxin [PCX]) were detected using Western blotting. PCX levels in extracellular vesicles were also detected using ELISA.

RESULTSTEM reveal the presence of numerous extracellular vesicles in the urine with intact morphology and different sizes, and most of them were below 300 nm in diameter as shown by NTA. TSG101 expression was detected in the samples from all the 4 groups. Positive expression of PCX was detected in the samples from patients with diabetic nephropathy but not in the other groups. In patients with diabetic nephropathy, the mean PCX levels (3.27±2.30 ng/μmol)was significantly higher than those in the healthy control group (1.22±0.36 ng/μmol), uncomplicated diabetes group (2.22±1.29 ng/μmol) and nephropathy group (1.24±0.45 ng/μmol).

CONCLUSIONSPCX level in urinary extracellular vesicles is significantly increased in patients with diabetic nephropathy, suggesting the value of PCX as a potential marker for clinical diagnosis of diabetic nephropathy.

PurposeTo measure the areas and diameter lines of bronchi at acute exacerbation and at remission period in patients with chronic obstructive pulmonary disease (COPD) using CT, and to explore the correlation between the two periods and evaluate the comprehensive assessment in diagnosing COPD exacerbation.Materials and Methods Fifty-two COPD patients were scanned with 64-row spiral CT on chest and PFT at acute exacerbation and at remission period. The areas and diameter lines of apical segmental and the sub-segmental bronchi of the right upper lobe in the patients were measured at the two periods, including indicators such as wall thickness (WT), thickness-diameter ratio (TDR), wall area (WA), percentage of wall area (WA%). The differences of those indicators at the two periods were compared with such factors of COPD comprehensive assessment as forced expiratory volume at the first second% (FEV1%), percentage of forced expiratory volume in first second to forced vital capacity (FEV1/FVC), COPD assessment test (CAT), modified medical research council questionnaire (mMRC) for assessing the severity of breathlessness, 6-minute walking distance (6MWD). Results The patients had significant differences between acute exacerbation period and remission period in the indicators of COPD comprehensive assessment like FEV1%, FEV1/FVC, CAT, mMRC and 6MWD (t=-4.119,-2.583, 4.012, 3.321 and-3.892,P<0.05). Compared with those at remission period, the WT, TDR, WA and WA% of sub-segmental bronchi were all higher at acute exacerbation period (t=3.025, 2.341, 2.204 and 2.124, P<0.05); only TDR of segmental bronchi showed significant difference between the two periods (t=2.990,P<0.05). The correlation of sub-segmental bronchi with FEV1%, FEV1/FVC, CAT, mMRC and 6MWD was more significant than that of segmental bronchi with those indicators at the two periods.Conclusion The COPD comprehensive assessment can help diagnose COPD at acute exacerbation period; MSCT shows the remodeling of segmental and sub-segmental bronchi and the changes on the airway wall, and the quantitative measurement of sub-segmental bronchi has correlation with the differences of indicators in the comprehensive assessment; COPD comprehensive assessment seems to be more valuable than PFT in the estimation of COPD at acute exacerbation.

OBJECTIVETo 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.

METHODSSixty-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.

RESULTSPatients 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.

CONCLUSIONIn the absence of renal biopsy findings, serum IgA/C3 ratio can help in the diagnosis of IgA nephropathy.

Biopsy ; Case-Control Studies ; Complement C3 ; analysis ; Glomerulonephritis, IGA ; blood ; diagnosis ; Humans ; Immunoglobulin A ; blood ; Kidney ; pathology

OBJECTIVETo analyze the quantity and size distribution of 24-hour urinary extracellular vesicles (uEVs) from healthy adults.

METHODSThe 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).

RESULTSuEVs 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.

CONCLUSIONThe 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.

Adult ; Blotting, Western ; Cell-Derived Microparticles ; Exosomes ; Extracellular Vesicles ; Humans ; Microscopy, Electron, Transmission ; Nanoparticles ; Urine

[ ABSTRACT ] AIM: To investigate the effect of advanced oxidation protein product-human serum albumin ( AOPP-HSA) at different concentrations on the permeability of human umbilical vein endothelial cell ( HUVEC) monolayer and the protective effect of NADPH oxidase inhibitor diphenyleneiodonium ( DPI ) against AOPP-HSA exposure. METHODS: Cultured HUVECs were exposed to 200 mg/L HSA (control) or AOPP-HSA (50, 100 and 200 mg/L).The permeability of the endothelial monolayer was assessed by measuring CMFDA-labeled THP-1 cells across the endothelial cells.The cultured HUVECs were treated with HSA (200 mg/L), AOPP-HSA (200 mg/L), or AOPP-HSA (200 mg/L)+DPI (100 μmol/L), and the activation of NADPH oxidase, endothelial monolayer permeability and cytoskeleton rear-rangement were evaluated.RESULTS: AOPP-HSA increased the permeability of the endothelial cell monolayer, and AOPP-HSA at 200 mg/L significantly increased the phosphorylation level of NADPH oxidase in the cells.Treatment with 100 μmol/L DPI obviously attenuated AOPP-HSA-induced NADPH oxidase activation, the increase in the permeability of the cell monolayer and the cytoskeleton rearrangement.CONCLUSION: AOPP-HSA increases the hyperpermeability of HUVEC monolayer via the phosphorylation of NADPH oxidase, and the NADPH oxidase inhibitor DPI reverses such effects.

Objective 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. Methods 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. Results 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. Conclusion In the absence of renal biopsy findings, serum IgA/C3 ratio can help in the diagnosis of IgA nephropathy.

Objective To investigate the prevalence of PLA2R1 in renal biopsy specimens of patients with idiopathic membranous nephropathy (IMN) and explore the relationship between PLA2R1 and IMN. Methods A total of 108 adult patients with biopsy-proved glomerular diseases were enrolled in this study, including 41 with IMN, 2 with hepatitis B-associated membranous nephropathy, 8 with V lupus nephritis, 27 with IgA nephropathy, 19 with minimal change nephropathy, 5 with mild mesangial proliferative glomerulonephritis, and 6 with focal segmental glomeruloselerosis (FSGS). Indirect immunofluorescence assay was used to detect PLA2R1 in the biopsy specimens and the clinical variables of the IMN patients were analyzed. Results In 35 of the 41 (85.37%) patients with IMN, PLA2R1 was detected with a fine granular pattern in the subepithelial deposits along the glomerular capillary loops. PLA2R1 antigen was not detected in patients with other glomerulopathies. No significant differences were found in age, serum creatinine, serum albumin, or 24-h urinary protein level between PLA2R1-positive and negative patients with IMN (P>0.05). Conclusion According to our results, 85.37% of adult patients with biopsy-proven IMN are positive for PLA2R1 antigen, which, however, does not contribute to variations of the patients' clinical manifestations.

Objective 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. Methods 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. Results 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. Conclusion In the absence of renal biopsy findings, serum IgA/C3 ratio can help in the diagnosis of IgA nephropathy.

Objective To investigate the prevalence of PLA2R1 in renal biopsy specimens of patients with idiopathic membranous nephropathy (IMN) and explore the relationship between PLA2R1 and IMN. Methods A total of 108 adult patients with biopsy-proved glomerular diseases were enrolled in this study, including 41 with IMN, 2 with hepatitis B-associated membranous nephropathy, 8 with V lupus nephritis, 27 with IgA nephropathy, 19 with minimal change nephropathy, 5 with mild mesangial proliferative glomerulonephritis, and 6 with focal segmental glomeruloselerosis (FSGS). Indirect immunofluorescence assay was used to detect PLA2R1 in the biopsy specimens and the clinical variables of the IMN patients were analyzed. Results In 35 of the 41 (85.37%) patients with IMN, PLA2R1 was detected with a fine granular pattern in the subepithelial deposits along the glomerular capillary loops. PLA2R1 antigen was not detected in patients with other glomerulopathies. No significant differences were found in age, serum creatinine, serum albumin, or 24-h urinary protein level between PLA2R1-positive and negative patients with IMN (P>0.05). Conclusion According to our results, 85.37% of adult patients with biopsy-proven IMN are positive for PLA2R1 antigen, which, however, does not contribute to variations of the patients' clinical manifestations.