Uria (mg/day) Serum albumin (g/dL) Hemoglobin (g/dL) Uric acid (mg/dL) FMD ( ) baPWV (cm/sec) Max IMT (mm) ACI ( ) 71 (62 ) 58 (51 ) 94613 9.3 (9.0?.5) 3.660.7 0.42 (0.26?.70) 16.0 (11.2?9.6) 50 (38?4) 15 (12?3) 35 (24?0) 44.4 (31.6?0.0) 616.3 (459.9?55.5) 119634 51 (41?4) 133 (89?83) 5.7 (5.5?.0) 0.05 (0.02?.15) 48629 439 (128?381) 3.9 (3.6?.2) 12.662.2 6.961.6 4.7 (3.1?.6) 1560 (1331?796) 0.85 (0.68?.10) 4.2 (0?6.4) 54 (47 ) 27 (24 ) 13 (11 ) 20 (18 ) 58 (47?6) 72 (59 )Results Patient characteristicsThe baseline characteristics of the study population are shown in Table 1. A total of 114 CKD patients with a median age of 58 (47?6) years were included in the study. The background causes of CKD included 54 cases of glomerulonephritis (47 ), 27 cases of nephrosclerosis (24 ), 13 cases of diabetic nephropathy (11 ) and 20 cases of “other” (18 ). A total of 83 patients were on antihypertensive therapy (71 patients were being treated with angiotensin receptor blockers (ARBs) or angiotensin converting enzyme inhibitors (ACEIs), 58 with calcium channel antagonistsACEI, angiotensin converting enzyme inhibitor; ACI, abdominal aortic calcification index; ARB, angiotensin receptor blocker; baPWV, brachial-ankle pulse wave velocity; CRP, C-reactive protein; 1,25D, 1,25-dihydroxyvitamin D; 25D, 25-hydroxyvitamin D; eGFR, estimated glomerular MedChemExpress Eliglustat filtration rate; FECa, fractional excretion of calcium; FEPi, fractional excretion of phosphate; FGF23, fibroblast growth factor 23; FMD, flow-mediated dilatation, HDL, high density lipoprotein; IMT, intima-media thickness; LDL, low density lipoprotein; MBP, mean blood pressure; NGSP, national glycohemoglobin standardization program. doi:10.1371/journal.pone.0056695.tSoluble Klotho and Arterial Stiffness in CKDRelationship between the serum Klotho level and age, renal function, CKD-related mineral metabolism and markers of vascular dysfunctionAge-dependent changes were recognized in the serum Klotho levels in patients with CKD (Figure 1A), as has been reported in healthy subjects [27]. The serum Klotho level was significantly correlated with the eGFR (Figure 1B) and decreased along with CKD stages (Figure S1A). With regard to markers of CKDMBD, the serum Klotho level was positively correlated with the 1,25-dihydroxyvitamin D (1,25D) level (Figure 1C) and negatively correlated with the log intact parathyroid hormone (PTH) and fractional excretion of phosphate (FEPi) (Figure 1D, 1E). The FEPi significantly increased along with declines in the eGFR (univariate regression, r = 20.7228, p,0.0001). There were no correlations between the level of serum Klotho and the fractional excretion of calcium (FECa) 1317923 (Figure 1F) or the 25-hydroxyvitamin D (25D) level (Figure S2C). However, correlations were observed between the level of serum Klotho and the level of serum calcium (r = 0.1618; p = 0.0855), the level of serum phosphate (r = 20.1454; p = 0.1426) and log intact FGF23 (r = 20.1751; p = 0.0624) (Figure S2A, Figure S2B and Figure S2D, respectively). We next investigated the association between the serum Klotho level and various markers of vascular dysfunction, including flowmediated dilatation (FMD), a marker of nitric oxide-dependent endothelial function, brachial-ankle pulse wave velocity (baPWV), a marker of arterial stiffness, maximum intima-media HIV-RT inhibitor 1 thickness (max IMT), a marker of atherosclerosis, and the abdominal aortic calcification index (ACI), a marker of vascular calcification (Figure 2). The serum Klotho lev.Uria (mg/day) Serum albumin (g/dL) Hemoglobin (g/dL) Uric acid (mg/dL) FMD ( ) baPWV (cm/sec) Max IMT (mm) ACI ( ) 71 (62 ) 58 (51 ) 94613 9.3 (9.0?.5) 3.660.7 0.42 (0.26?.70) 16.0 (11.2?9.6) 50 (38?4) 15 (12?3) 35 (24?0) 44.4 (31.6?0.0) 616.3 (459.9?55.5) 119634 51 (41?4) 133 (89?83) 5.7 (5.5?.0) 0.05 (0.02?.15) 48629 439 (128?381) 3.9 (3.6?.2) 12.662.2 6.961.6 4.7 (3.1?.6) 1560 (1331?796) 0.85 (0.68?.10) 4.2 (0?6.4) 54 (47 ) 27 (24 ) 13 (11 ) 20 (18 ) 58 (47?6) 72 (59 )Results Patient characteristicsThe baseline characteristics of the study population are shown in Table 1. A total of 114 CKD patients with a median age of 58 (47?6) years were included in the study. The background causes of CKD included 54 cases of glomerulonephritis (47 ), 27 cases of nephrosclerosis (24 ), 13 cases of diabetic nephropathy (11 ) and 20 cases of “other” (18 ). A total of 83 patients were on antihypertensive therapy (71 patients were being treated with angiotensin receptor blockers (ARBs) or angiotensin converting enzyme inhibitors (ACEIs), 58 with calcium channel antagonistsACEI, angiotensin converting enzyme inhibitor; ACI, abdominal aortic calcification index; ARB, angiotensin receptor blocker; baPWV, brachial-ankle pulse wave velocity; CRP, C-reactive protein; 1,25D, 1,25-dihydroxyvitamin D; 25D, 25-hydroxyvitamin D; eGFR, estimated glomerular filtration rate; FECa, fractional excretion of calcium; FEPi, fractional excretion of phosphate; FGF23, fibroblast growth factor 23; FMD, flow-mediated dilatation, HDL, high density lipoprotein; IMT, intima-media thickness; LDL, low density lipoprotein; MBP, mean blood pressure; NGSP, national glycohemoglobin standardization program. doi:10.1371/journal.pone.0056695.tSoluble Klotho and Arterial Stiffness in CKDRelationship between the serum Klotho level and age, renal function, CKD-related mineral metabolism and markers of vascular dysfunctionAge-dependent changes were recognized in the serum Klotho levels in patients with CKD (Figure 1A), as has been reported in healthy subjects [27]. The serum Klotho level was significantly correlated with the eGFR (Figure 1B) and decreased along with CKD stages (Figure S1A). With regard to markers of CKDMBD, the serum Klotho level was positively correlated with the 1,25-dihydroxyvitamin D (1,25D) level (Figure 1C) and negatively correlated with the log intact parathyroid hormone (PTH) and fractional excretion of phosphate (FEPi) (Figure 1D, 1E). The FEPi significantly increased along with declines in the eGFR (univariate regression, r = 20.7228, p,0.0001). There were no correlations between the level of serum Klotho and the fractional excretion of calcium (FECa) 1317923 (Figure 1F) or the 25-hydroxyvitamin D (25D) level (Figure S2C). However, correlations were observed between the level of serum Klotho and the level of serum calcium (r = 0.1618; p = 0.0855), the level of serum phosphate (r = 20.1454; p = 0.1426) and log intact FGF23 (r = 20.1751; p = 0.0624) (Figure S2A, Figure S2B and Figure S2D, respectively). We next investigated the association between the serum Klotho level and various markers of vascular dysfunction, including flowmediated dilatation (FMD), a marker of nitric oxide-dependent endothelial function, brachial-ankle pulse wave velocity (baPWV), a marker of arterial stiffness, maximum intima-media thickness (max IMT), a marker of atherosclerosis, and the abdominal aortic calcification index (ACI), a marker of vascular calcification (Figure 2). The serum Klotho lev.
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