Yi-kuan Chen, MD, Xue-mei Jiang, MSc, Jian-ping Gong, MD

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Recombinant human granulocyte colony-stimulating factor enhanced the resolution of venous thrombi Yi-kuan Chen, MD, Xue-mei Jiang, MSc, Jian-ping Gong, MD Journal of Vascular Surgery Volume 47, Issue 5, Pages 1058-1065 (May 2008) DOI: 10.1016/j.jvs.2007.12.042 Copyright © 2008 The Society for Vascular Surgery Terms and Conditions

Fig 1 Organization of thrombi in thrombus group (ThG) and treatment group (TrG) at postoperative day 10 (%, n = 10). The red arrows indicate the organization area of thrombi. As organization of venous thrombi occurred, neutrophils, monocytes, and endothelial cells entered into the thrombus, neovessels appeared, and thrombus collagen formed. The organization rate was significantly different in the treatment group (80.2% ± 10.8%; P < .01) compared with the thrombus group (30.6% ± 3.5%). A, Organization of thrombi in the thrombus group was mainly located at the peripheral area of thrombi. B, Organization of thrombi in the treatment group involved almost all areas of the thrombi (hematoxylin and eosin [HE], original magnification ×40). C, Few neovessels appeared at the organization area of thrombus in the thrombus group. D, Many neovessels formed at the organization area of thrombus in the treatment group, and the neovessels coalesced and enlarged (HE original magnification ×100). Journal of Vascular Surgery 2008 47, 1058-1065DOI: (10.1016/j.jvs.2007.12.042) Copyright © 2008 The Society for Vascular Surgery Terms and Conditions

Fig 2 Immunohistochemical CD68 stain of thrombi in thrombus group (ThG) and treatment group (TrG) at postoperative day 10 (Streptavidin-biotin-peroxidase complex, original magnification ×400). Macrophages stained brown and are indicated by the red arrows. A, A few macrophages at vein wall (W) and thrombus (T) in the thrombus group. Error bars show the standard error of the mean. B, Many macrophages and neovessels are seen at the thrombus in the treatment group. The neovessels mostly appeared at areas where macrophages accumulated. The macrophage number in the thrombus was significantly increased in the treatment group (338 ± 26 cells/15 high-power fields) compared with the thrombus group (125 ±11 cells/15 high-power fields; P < .01). Journal of Vascular Surgery 2008 47, 1058-1065DOI: (10.1016/j.jvs.2007.12.042) Copyright © 2008 The Society for Vascular Surgery Terms and Conditions

Fig 3 Expression of the cysteine-cysteine chemokine receptor 2 (CCR2) gene in the peripheral blood of the four experimental groups. Real-time reverse transcriptase–polymerase chain reaction of each transcript was performed. Results are presented as relative units of each CCR2 transcript compared with glyceraldehyde-3-phosphate dehydrogenase (GAPDH). NG, Normal (control) group; SG, sham operation group; ThG, thrombus group; TrG, treatment group. #P < .05 compared with thrombus group. Error bars show the standard error of the mean. Journal of Vascular Surgery 2008 47, 1058-1065DOI: (10.1016/j.jvs.2007.12.042) Copyright © 2008 The Society for Vascular Surgery Terms and Conditions

Fig 4 Left, Cysteine-cysteine chemokine receptor 2 (CCR2) protein expression of TPH-1 cells treated with different concentration of recombinant human granulocyte colony-stimulating factor (rhG-CSF) was determined by Western blot analysis for 48 hours. Right, Graph shows combined results from three culturing flasks. Lanes 1, 2, 3, 4, 5, 6 and 7 were the markers and 0, 20, 40, 80, 160 and 320 ng/mL was the respective concentration of rhG-CSF. GAPDH, Glyceraldehyde-3-phosphate dehydrogenase. *P < .05, #P < .01 compared with 0 ng/mL. Error bars show the standard error of the mean. Journal of Vascular Surgery 2008 47, 1058-1065DOI: (10.1016/j.jvs.2007.12.042) Copyright © 2008 The Society for Vascular Surgery Terms and Conditions

Fig 5 Left, Expression of cysteine-cysteine chemokine receptor 2 (CCR2) protein in 80 ng/mL recombinant human granulocyte colony-stimulating factor treated TPH-1 cells as determined by Western blot analysis for 36, 48 and 60 hours. GAPDH, Glyceraldehyde-3-phosphate dehydrogenase. Right, Graph shows the combined results from three culturing flasks. Compared with 36 hours of culture, there was significant difference in expression of CCR2 protein from 48 hours. #P < .01. Error bars show the standard error of the mean. Journal of Vascular Surgery 2008 47, 1058-1065DOI: (10.1016/j.jvs.2007.12.042) Copyright © 2008 The Society for Vascular Surgery Terms and Conditions