Association with inflammatory cells and apolipoproteins to the progression of atherosclerosis.

PURPOSE: Inflammatory cells are known to be associated with the progression of atherosclerosis and plaque rupture. However, the relation to inflammatory cells and apolipoproteins on the progression of atherosclerosis is unknown. This study was aimed at examining the different expressions of inflammatory cells and evaluate the effect of apolipoprotein (APO) C1 and APO E during the progression of atherosclerosis. METHODS: Ten atherosclerotic tissues were compared with five non-atherosclerotic tissues. The presence of vascular smooth muscle cells (VSMCs), macrophages, T-cells, APO C1, and APO E were identified by Western blotting and immunohistochemical analysis with antibodies. The senescence was analyzed by senescence-associated ß-galactosidase. RESULTS: The protein expression and senescence of macrophages, APO C1 and APO E were significantly higher in the main atherosclerotic lesion than the non-atherosclerotic lesion. A high concentration of inflammatory cells and the paucity of VSMCs were present in the shoulder area. In addition, macrophage and T-cells are expressed in the early stage of atherosclerotic development and more expanded in advanced atherosclerotic plaques. APO C1 was expressed mainly within the necrotic core, and APO E existed mostly around the necrotic core and the fibrous cap in advanced atherosclerotic plaques. CONCLUSION: Our study indicated that the expression and the senescence of macrophage and T-cells may be closelyrelated to induction and deposition of APO C1 and APO E. This contributes to the development and progression of atherosclerotic plaque by expanding the necrotic core.

Apolipoprotein C1 and apolipoprotein E are differentially expressed in atheroma of the carotid and femoral artery.

BACKGROUND: A number of the genes and proteins as the causes of carotid atherosclerotic disease have been recently reported, but the major factors for atherosclerosis have still not been identified. METHODS: The atherosclerotic atheromas were obtained during endarterectomy for each of 10 cases of diseased carotid and femoral arteries. As the nonatherosclerotic arteries, the iliac arteries were obtained during organ harvest from five cases of brain-dead donors, and the leg arteries were obtained during leg amputation from five cases of Buerger's disease. The total RNAs and proteins were isolated from the atheromas and arteries. The annealing control primer method was used to screen the differentially expressed mRNAs. To identify if the mRNA expression of screened gene was associated with the protein expression, we performed an immunohistochemical analysis. RESULTS: We found that the apolipoprotein C1 (apo C1) gene was prominently expressed in the atheroma of the carotid and femoral arteries, as compared to the nonatherosclerotic arteries. Immunohistochemical analysis showed the high expression of apo C1 protein in the atheromas of the carotid and femoral arteries. Apo E protein was also highly expressed in atheromas compared with the nonatherosclerotic arteries, but there was no difference for apo C2 protein between those four groups of arteries. DISCUSSION: The expression of apo C1 and apo E are closely associated with the susceptibility to the pathogenesis of atherosclerosis. This study suggests that these factors might play important roles in the future to screen for preventing atherosclerosis and for diagnostic testing of patients.

Angiogenesis facilitated by autologous whole bone marrow stem cell transplantation for Buerger's disease.

We hypothesized that angiogenesis can be triggered by autologous whole bone marrow stem cell transplantation. Twenty-seven patients (34 lower limbs) with Buerger's disease, who were not candidates for surgical revascularization or radiologic intervention, were enrolled in this study. Six sites of the tibia bone were fenestrated using a 2.5-mm-diameter screw under fluoroscopic guidance. Clinical status and outcome were determined using the "Recommended Standards for Reports." To mobilize endothelial progenitor cells (EPCs) from bone marrow, recombinant human granulocyte colony-stimulating factor (r-HuG-CSF) was injected subcutaneously as a dose of 75 microg, preoperatively. During the follow-up period (19.1 +/- 3.5 months), one limb showed a markedly improved outcome (+3), and 26 limbs showed a moderately improved outcome (+2). Thirteen limbs among 17 limbs with nonhealing ulcers healed. Postoperative angiograms were obtained for 22 limbs. Two limbs showed marked (+3), five limbs moderate (+2), and nine limbs slight (+1) collateral development. However, six limbs showed no collateral development (0). Peripheral blood and bone marrow samples were analyzed for CD34 and CD133 molecules to enumerate potential EPCs, and EPC numbers were found to be increased in peripheral blood and in bone marrow after r-HuG-CSF injection. In conclusion, the transplantation of autologous whole BMCs by fenestration of the tibia bone represents a simple, safe, and effective means of inducing therapeutic angiogenesis in patients with Buerger's disease.

Differential expression of immunoglobulin kappa chain constant region in human abdominal aortic aneurysm.

BACKGROUND: A number of the research into the pathogenesis of the abdominal aortic aneurysm (AAA) have focused on the alteration of gene expression. The current technique for elucidating alterations of gene expression has a setback in that many artifact complementary DNA (cDNA) products present abnormal polymerase chain reaction (PCR) amplification. Our study was designed to identify differentially expressed genes in AAA using the annealing control primer (ACP) system, which was recently developed to identify only authentic genes. MATERIALS AND METHODS: The tissues of the human abdominal aorta were obtained from the patients of AAA and aortic occlusive disease (AOD), and normal abdominal aorta (NA) from brain death donors. Total RNAs were isolated from three groups of human abdominal aorta (10 AAA, five NA, three AOD) and then reverse transcribed into complementary DNA (cDNA). The ACP method was done to screen the difference in the expression pattern of the mRNA (mRNA). RESULTS: One differentially expressed cDNA band was detected in AAA but not in NA and AOD. This cDNA was sequenced and computer searching against the GenBank revealed that the cDNA had more than 90% identity with the immunoglobulin kappa chain constant region (Ig kappa-C). DISCUSSION: Our finding suggests that differentially expressed Ig kappa-C gene only in AAA is a candidate gene that may play a pivotal role in the pathogenesis of AAA formation. The correlation of mRNA level and protein level is, however, not clear. Thus, to directly identify the role of Ig light chains in the pathogenic event of AAA, the further study comparing the level and kinds of expressed protein with the corresponding Ig kappa-C gene will be required.

Identification of differentially expressed genes in primary varicose veins.

BACKGROUND: A number of changes in protein expression have been described in primary varicose veins, but the altered gene expressions in this disease are unknown. The aim of this study was to identify differentially expressed genes in primary varicose veins. MATERIALS AND METHODS: Total RNAs were isolated from two groups of greater saphenous veins (four primary varicose veins and three normal) and then were reverse transcribed into cDNAs. We used the differential display reverse transcription-polymerase chain reaction technique to screen the differences in the mRNA expression profiles of the groups. RESULTS: We found that three cDNAs showed differences in expression patterns between normal and diseased saphenous veins. The cDNAs are prominently expressed only in patients with varicose veins. We identified that the cDNAs had significant similarities to the L1M4 repeat sequence of clone RP11-57L9, clone RP11-299H13, and Alu repetitive sequence of human tropomyosin 4 mRNA. CONCLUSIONS: Our results suggest that the screened cDNA clones are useful disease markers in the genetic diagnosis of primary varicose vein and that the L1 and Alu elements possibly participated in the development of primary varicose veins through their expression patterns in genes encoded with structural proteins, such as collagen, elastin, and tropomyosin. Further studies are required to elucidate the potential relationship between repeat sequences and primary varicose veins.