High-mobility-group box protein 1 A box reduces development of sodium laurate- induced thromboangiitis obliterans in rats Xiangqian Kong, MD, Hai Yuan,

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High-mobility-group box protein 1 A box reduces development of sodium laurate- induced thromboangiitis obliterans in rats Xiangqian Kong, MD, Hai Yuan, MD, Xuejun Wu, MD, Jingyong Zhang, MD, Hua Zhou, MD, Maohua Wang, MD, Yang Liu, MD, Xing Jin, MD Journal of Vascular Surgery Volume 57, Issue 1, Pages 194-204 (January 2013) DOI: 10.1016/j.jvs.2012.06.083 Copyright © 2013 Society for Vascular Surgery Terms and Conditions

Fig 1 A, Gross appearance of rat paws; (B) hematoxylin and eosin staining of femoral arteries (original magnification, ×100) in (a) normal (nor), (b) sham-operated (sham), (c) thromboangiitis obliterans (TAO) model, and (d) low-dose (rA box-LD) and (e) high-dose (rA box-LD) recombinant A (rA) box-treated groups; and (C) data for the area ratio of thrombus of femoral arteries on day 15 after sodium laurate injection. Data are shown as mean ± standard error of the mean (error bars). **P < .01 vs sham-operated group. #P < .05 vs TAO model group. Journal of Vascular Surgery 2013 57, 194-204DOI: (10.1016/j.jvs.2012.06.083) Copyright © 2013 Society for Vascular Surgery Terms and Conditions

Fig 2 A, Plasma levels of high-mobility-group box protein 1 (HMGB1) are shown for normal (nor), sham-operated (sham), and thromboangiitis obliterans (TAO) model and the low-dose (rA box-LD) and high-dose (rA box-HD) recombinant A (rA) box-treated groups. Results are means ± standard error of the mean (error bars); n = 8. **P < .01 vs sham-operated group. #P < .05 vs TAO model group. B, Representative micrographs of HMGB1 in rat femoral arteries by 3,3-diaminobenzidine staining and (C) double immunofluorescent staining for receptor for advanced glycation end-products (RAGE) and cluster of differentiation 31 (CD 31) are shown for (a) nor, (b) sham, (c) TAO, and (d) rA box-LD and (e) rA box-HD groups. Journal of Vascular Surgery 2013 57, 194-204DOI: (10.1016/j.jvs.2012.06.083) Copyright © 2013 Society for Vascular Surgery Terms and Conditions

Fig 3 Western blot and reverse transcription polymerase chain reaction analysis of protein and messenger RNA levels of high-mobility-group box protein 1 (HMGB1) and receptor for advanced glycation end-products (RAGE) in femoral arteries from normal (nor), sham-operated (sham), thromboangiitis obliterans (TAO) model, and low-dose (rA box-LD) and high-dose (rA box-HD) recombinant A (rA) box-treated rats. A, Representative Western blot of protein levels of HMGB1 and RAGE (β-actin as a control). B, Summary data for the rat groups are presented as means ± standard error of the mean (error bars); n = 8. **P < .01 vs sham-operated group. #P < .05 vs TAO model group. C, The relative messenger RNA levels of HMGB1 and RAGE are presented as means ± standard error of the mean (error bars); n = 8. **P < .01 vs sham-operated group. #P < .05 vs TAO model group. Journal of Vascular Surgery 2013 57, 194-204DOI: (10.1016/j.jvs.2012.06.083) Copyright © 2013 Society for Vascular Surgery Terms and Conditions

Fig 4 Immunohistochemistry, Western blot, and reverse transcription polymerase chain reaction analysis of protein and messenger RNA levels of interleukin-6 (IL-6), intercellular adhesion molecule-1 (ICAM-1), and vascular cell adhesion molecule-1 (VCAM-1) in femoral arteries from (a) normal (nor), (b) sham-operated (sham), (c) thromboangiitis obliterans (TAO) model, and (d) low-dose (rA box-LD) and (e) high-dose (rA box-HD) recombinant A (rA) box-treated rats. A, Representative photomicrographs of IL-6, ICAM-1, and VCAM-1 in rat femoral arteries by 3,3-diaminobenzidine staining. B, Representative Western blot of protein levels of IL-6, ICAM-1, and VCAM-1 (β-actin as a control). C, Summary data for rat groups presented as means ± standard error of the mean (error bars); n = 8. *P < .05 vs sham-operated group. **P < .01 vs sham operated group. #P < .05 vs TAO model group. ##P < .01 vs TAO model group. D, The relative messenger RNA levels of IL-6, ICAM-1, and VCAM-1 for rat groups are presented as means ± standard error of the mean (error bar); n = 8. *P < .05 vs sham operated group. **P < .01 vs sham operated group. #P < .05 vs TAO model group. Journal of Vascular Surgery 2013 57, 194-204DOI: (10.1016/j.jvs.2012.06.083) Copyright © 2013 Society for Vascular Surgery Terms and Conditions

Fig 5 Plasma levels of thromboxane B2 (TXB2) and 6-keto-prostaglandin F1-α (6-K-PGF1-α) and the TXB2/6-K-PGF1α ratio are shown in normal (nor), sham-operated (sham), thromboangiitis obliterans (TAO) model, and low-dose (rA box-LD) and high-dose (rA box-HD) recombinant A (rA) box-treated group. Results are expressed in means ± standard error of the mean (error bars); n = 8. *P < .05 vs sham operated group. **P < .01 vs sham operated group. #P < .05, vs TAO model group. Journal of Vascular Surgery 2013 57, 194-204DOI: (10.1016/j.jvs.2012.06.083) Copyright © 2013 Society for Vascular Surgery Terms and Conditions