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Table 1.

General characteristics of study subjects.

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Table 2.

Correlation between DR and laboratory characteristics.

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Table 3.

Correlation between DR and CTRPs.

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Fig 1.

ROC curves, for DR/PDR diagnosis, by circulating CTRP3 level.

(A) ROC curve analysis revealed that CTRP3 distinguishes patients with and without DR, AUC = 0.900 (95% CI 0.838–0.962, P<0.001). (B) CTRP3 distinguishes PDR from NPDR, AUC = 0.919 (95% CI 0.850–0.989, P<0.001).

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Table 4.

Correlation analysis of variables associated with circulating CTRP3.

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Fig 2.

CTRP3 inhibits high glucose/high lipids (HGHL)-induced expression of VCAM-1 in a time- and dose-dependent manner.

(A) HRMECs were subjected to high glucose/high lipid endothelial growth medium for 48 hours, incubated with CTRP3 (0.3, 1, 3μg/mL) for 60 minutes. VCAM-1 was inhibited in a concentration-dependent manner. (B) HRMECs were subjected to various periods of CTRP3 treatment (30, 60, and 120 minutes). VCAM-1 significantly decreased after 15 minutes, with decreasing trend for 120 total minutes. (C) HRMECs were placed in transwell migration chambers containing filters coated with matrigel. To stimulate migration, different concentrations of CTRP3 (0, 0.3, 1, 3μg/ml) were added to the lower chamber. Transwell chambers were stained with crystal violet and imaged after 8 hours. (D) The average number of migrated cells was quantified compared to vehicle control. Results are expressed as means±SD from 5–6 independent experiments. Each experiment was repeated 3 times. *, P<0.05; ##, P<0.01 vs respective control.

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Fig 3.

CTRP3 inhibited HGHL induced VCAM-1 production in an AMPK dependent manner.

(A) HRMECs were incubated with HGHL for 48 hours, and subjected to CTRP3 administration (doses 0.3, 1, and 3μg/ml). (B) Akt phosphorylation was determined after 15 minutes. (C) AMPK phosphorylation was determined after 48 hours of HGHL incubation, followed by 15 minutes of CTRP3 treatment (doses 0.3, 1, and 3μg/ml). (D) Western blot analysis confirmed successful knockdown of AMPK by siRNA (>80% knockdown) E) AMPK knockdown blocked CTRP3 (3μg/ml, 1 hour treatment)-mediated inhibition of VCAM-1 expression induced by HGHL (48 hours treatment). Results are expressed as means±SD from 5–6 independent experiments. Each experiment was repeated 3 times. *p<0.05; ##, p<0.01 vs respective control.

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