Receiver operating characteristic (ROC) curve analysis revealed the optimal cut-off points for Siglec-5 was 153

Receiver operating characteristic (ROC) curve analysis revealed the optimal cut-off points for Siglec-5 was 153.1 ng/ml. the optimal cut-off points for Siglec-5 was 153.1 ng/ml. After entering Siglec-5, the AUC was 0.99, which was higher than that of confounding risk factors only (AUC?=?0.97, non-CLI control plasma) and statistical significance of the difference (both expressed in log scales). (B) Proteins that exhibited significant increases or decreases in CLI patients ( em n /em ?=?4) compared with non-CLI patients ( em n /em ?=?4) in the antibody array analysis, where the CLI patients and non-CLI ( em n /em ?=?4) patients are represented as black and white bars, respectively, * em P /em ? ?0.05. Confirmation study of biomarkers for CLI in T2DM ELISA analysis results of the eight samples tested in protein arrays yielded results were consistent with that of the protein arrays. We then performed a confirmation study in an independent cohort of T2DM patients with CLI ( em n /em ?=?59) and without CLI ( em n /em ?=?108) by ELISA. As shown in Table?2 and Fig.?2, among the 11 potential biomarkers, seven (BMP-4, IGFBP-6, Siglec-5, Decorin, Trappin-2, TIMP-4, and CDH5) were significantly elevated in the CLI group compared with the non-CLI group, while there were no significant differences in the levels of NCAM-1, MSP-alpha, Axl or EpCAM. Table 2 Confirmation assays of plasma proteins in diabetes and their potential to discriminate lower limb sclerosis from controls (pg/ml). thead th rowspan=”2″ colspan=”1″ Proteins /th th colspan=”2″ rowspan=”1″ protein array cohort? /th th colspan=”2″ rowspan=”1″ confirmation cohort /th th rowspan=”1″ colspan=”1″ CLI ( em n /em ?=?4) /th th rowspan=”1″ colspan=”1″ Non-CLI ( em n /em ?=?4) /th th rowspan=”1″ colspan=”1″ CLI ( em n /em ?=?59) /th th rowspan=”1″ colspan=”1″ Non-CLI ( em n /em ?=?108) /th /thead BMP-4497.34??26.24# 385.74??20.66558.57??34.20# 437.14 (330.11C544.50)? IGFBP-6209.06??21.49# 99.93??17.95195.62??19.59# 124.14 (88.49C167.22)? NCAM-1(182.92??11.42)*10?3# (277.58??18.64)*10?3 (264.20??8.25)*10?3 (264.40??7.11)*10?3 MSP-alpha57.34??6.99# 31.60??3.1459.30??3.2158.79??2.13Siglec-5(258.76??37.21)*103# (45.66??6.00)*103 (193.62 (158.08C196.32))*103### (73.84??4.76)*103? Axl14.64??1.48# 26.71??1.6423.33??1.3621.37 (13.74C29.02)? Decorin(1.90??0.30)*103# (3.52??0.16)*103 (3.75??0.18)*103## (3.14??0.12)*103 Trappin-233.42??2.78# 14.08??2.4923.22 (13.23C50.34)### 13.22 (9.43C19.02)? EpCAM10.23??0.23# 5.00??1.257.30??0.686.50 (1.38C10.88)? TIMP-4(4.19??0.30)*103# (2.08??0.21)*103 (3.94??0.41)*103### (2.18??0.22)*103 CDH5(40.26??1.77)*103# (29.47??1.59)*103 (37.92??1.23)*103### (32.92??0.56)*103 Open in a separate window Data is shown as mean??SEM, median (interquartile range) of subjects in each group. Differences between the groups in the protein array cohort were analyzed using ?) Fisher-Pitman Permutation test, however, the confirmation cohort was analyzed using ?) Mann-Whitney U-test, all the others were analyzed using em Eupalinolide B t /em -tests. # em p /em ? ?0.05; ## em p /em ? ?0.01; ### em p /em ? ?0.001. Open in a separate window Figure 2 A confirmation study for the findings of antibody array analysis (11 potential proteins) in an independent cohort subjects with CLI ( em n /em ?=?59) and non-CLI ( em n /em ?=?108) patients by ELISA. The plasma levels of the 11 proteins in the respective study groups. BMP-4, IGFBP-6, Siglec-5, Decorin, Trappin-2, TIMP-4, and CDH5 were significantly elevated in CLI compared with non-CLI (* em P /em ? ?0.05). Analysis of plasma cytokine levels and CLI risk factors With the exception of BMP-4, the plasma cytokine levels increased with age, T2DM duration or BUN (all em Eupalinolide B P /em ? ?0.05). The plasma levels of cytokines (Decorin, Siglec-5 and TIMP-4) increased with SBP (all em P /em ? ?0.05). There were negative associations between TC and plasma Siglec-5 or TIMP-4 levels (all em P /em ? ?0.05). There were also negative associations between Apo A and plasma Trappin-2, Siglec-5 or CDH5 levels (all em P /em ? ?0.05). With the exception of BMP-4 and Decorin, the plasma cytokine levels increased with Cr (all em P /em ? ?0.05). In addition, increased plasma levels of Trappin-2, IGFBP-6, Siglec-5 and TIMP-4 correlated with lower total bilirubin levels (all em P /em ? ?0.05) (Table?3). Table 3 Spearman Rho correlations analysis of plasma cytokine levels and CLI risk factors. thead th align=”left” rowspan=”1″ colspan=”1″ Variable /th th align=”left” rowspan=”1″ colspan=”1″ /th th align=”left” rowspan=”1″ colspan=”1″ Trappin-2 /th th align=”left” rowspan=”1″ colspan=”1″ IGFBP-6 /th th align=”left” rowspan=”1″ colspan=”1″ BMP-4 /th th align=”left” rowspan=”1″ colspan=”1″ Decorin /th th align=”left” rowspan=”1″ colspan=”1″ siglec-5 /th th align=”left” rowspan=”1″ colspan=”1″ TIMP-4 /th th align=”left” rowspan=”1″ colspan=”1″ CDH5 /th /thead Age em rho /em 0.4210.2760.1360.3560.5170.5070.224 em p /em em 2 /em . em 06E-8 /em em 3 /em . em 44 E-4 /em 0.085 em 4 /em . em 75E-6 /em em 8 /em . em 75E-13 /em em 4 /em . em 13E-6 /em em 0 /em . em 005 /em T2D Duration em rho /em 0.3330.3310.0980.2310.2670.4060.187 em p /em em 1 /em . em 34E-5 /em em 1 /em . em 49E-5 /em 0.216 em 0 /em . em 004 /em em 4 /em . em 76E-4 /em em 3 /em . em 30E-4 /em em 0 /em . em 018 /em SBP em rho /em 0.0720.054?0.0760.1860.3060.3010.104 em p /em 0.3560.4960.335 em 0 /em . em 019 /em em 5 /em . em 65E-5 /em em 0 /em . em 009 /em 0.192TC em rho /em ?0.150?0.0430.121?0.006?0.170?0.278?0.137 em p /em 0.0640.5950.1370.940 em 0 /em . em 033 /em em 0 /em . em 021 /em 0.097Apo A em rho /em ?0.2380.0460.0140.028?0.186?0.117?0.174 em p /em em 0 /em . em 003 Eupalinolide B /em 0.5740.8610.738 em 0 /em . em 021 /em 0.349 em 0 /em . em 037 /em Cr em rho /em 0.5200.4970.0150.0750.2140.4220.187 em P /em em 3 /em . em 36E-12 /em em 4 /em . em 07E-11 /em 0.8490.361 em 0 /em . em 007 /em em 3 /em . em 05E-4 /em em 0 /em . em 022 /em BUN em rho /em 0.3760.3480.0140.1850.3050.3430.187 em p /em Rabbit Polyclonal to OR5P3 em 1 /em . em 23E-6 /em em 7 /em . em 99E-6 /em 0.867 em 0 /em . em 023 /em em 8 /em . em 61E-5 /em em 0 /em . em 004 /em em 0 /em . em 021 /em Total bilirubin em rho /em ?0.288?0.208?0.133?0.073?0.309?0.317?0.137 em p /em em 2 /em . em 53E-4 /em em 0 /em . em 009 /em 0.0990.372 em 7 /em . em 07E-3 /em em 0 /em . em 007 /em 0.093 Open in a separate window All study subjects were included in the analysis. Significant values are marked in italic. Risk factors for CLI Taking CLI as the dependent variable, the risk factors shown in Table?1 were entered into binary logistic regression analysis. The significant risk factors included sex, age,.