Abstract: Objective To induce calcification in aortic valvular interstitial cells (VICs) in vitro and observe the shift of cellular phenotype during the process. Methods Porcine aortic VICs were isolated and expanded by collagenase methods. Fluorescent staining was performed to identify the interstitial cells. VICs at 48 passages were used for experiments. The cells were divided into two groups: the experimental group in which cells were cultured in osteogenic media supplemented with βglycerophosphate, vitamin C and dexamethasone, and the control group in which cells were cultured in normal media. After 2 weeks, calcified nodules were quantified. Calcium deposit was stained and measured by Alizarin Red S staining and assay. Real time reverse transcription polymerase chain reaction (RTPCR) was performed to measure expression of alpha smooth muscle actin (α-SMA) and calcification related factors such as osteocalcin, osteopontin and Corebinding factor α1/Runx2 (Cbfα1/Runx2). Results VICs were successfully harvested from porcine aortic valves, identified by positive staining of α-SMA, vimentin and negative staining of Von Willebrand factor (vWF). VICs could calcify after 2 weeks of osteogenic induction with calcified nodules formed. Quantification of calcified nodules and calcification deposit were significantly higher (Plt;0.05) in the experimental group than those in the control group (156.25±17.38 vs. 2.50±1.29, 17.52±2.04 vs. 1.00±0.22). Real Time RT-PCR indicated that expression of α-SMA, as well as calcification related markers like osteocalcin, osteopontin and Cbfα1/Runx2 was much higher in the experimental group than those in the control group (Plt;0.05). Conclusion VICs are activated during the progress of calcification with phenotype shifting to contraction and ossification, which might be the pathological basis of valvular calcification.
【摘要】目的 探讨胸部X线片检查发现的主动脉弓钙化与冠心病的不同临床表现类型的相关性。方法 回顾分析2006年7月-2008年2月期间经冠状动脉造影证实的冠心病患者的临床资料,对比分析胸部平片所见的主动脉弓钙化情况与冠心病不同临床类型的相互关系。结果 116例经冠状动脉造影证实的冠心病患者纳入研究。其中,稳定型心绞痛40例,急性冠脉综合征76例(不稳定型心绞痛21例、非ST段抬高心肌梗死38例、ST段抬高心肌梗死17例)。40例稳定型心绞痛患者中,有主动脉弓钙化者21例,占52.5%;76例急性冠脉综合征患者中,有主动脉弓钙化者22例,占28.9%。与急性冠脉综合征相比,更多的稳定型心绞痛患者合并有主动脉弓钙化(χ2=6232,P=0013)。结论 在不同的冠心病临床类型,主动脉弓钙化更易在稳定型心绞痛患者中发现。【Abstract】 Objective To investigate the relationships between calcification of aortic arch and different clinical classification of coronary artery disease. Methods The clinical data of patients with coronary artery diease who diagnosed by arteriography from July 2006 to February 2008, were retrospectively analyzed. The Xray data on calcification of aortic arch and clinical characteristics of patients with coronary artery disease confirmed by coronary angiography were analyzed. The relationship between coronary calcification of aortic arch which showed by Xray and different clinical classification of coronary artery disease were comparatively analyzed. Results Among the total of 116 patients, 40 stable angina and 76 acute coronary syndrome were included, and 21 (52.5%) and 22 (28.9%) patients with calcification of aortic arch were observed respectively. In comparison to patients with acute coronary syndrome, more stable angina patients were complicated with calcification of aortic arch (χ2 =6232,P=0013). Conclusion It is more likely to document calcification of aortic arch in patients with stable angina.
ObjectiveTo investigate the X-ray diagnostic significance of calcification of the breast tumor without mass. MethodsMammograms of 90 cases of breast tumor without mass confirmed pathologically were retrospectively analyzed. There were 55 cases confirmed benign breast tumor, and the rest cases were breast cancer. The shape, distribution, total number, location of calcifications in the breast, and asymmetric dense of the breast were recorded and watched. Results①The X-ray findings of calcification in benign breast tumors always presented as coarse granular (31), scattered shape (35) with small number, less with the asymmetric dense of the breast (7), and the change of side with axillary lymph node (2). ②Meanwhile, fine sand-like (32), showing the cluster-like distribution (24) with larger number, with the asymmetric dense of the breast (24) and the change of side with axillary lymph node (10). Both of the differences of the calcifications (the shape, the distribution, and the total number) were statistically significant (Plt;0.05). ConclusionsThe calcifications of benign and malignant breast tumors have their unique X-ray characteristics. And there is a great value in differentiating early benign and malignant breast tumor.
ObjectiveExtracting the endothelial cells or all endothelial cells and interstitial cells from the cryopreserved homograft valves (HV), to evaluate the immunogenicity of this two kinds of decellular HV. MethodsFor extracting the endothelial cells, the leaflet and wall of the HV were decellularized by a 4-step detergent-enzymatic extraction method involving the 1% triton in combination with RNase (1μg/ml) and DNase (10μg/ml). For extracting the endothelial cells and interstitial cells, the leaflet and wall of the HV were decellularized by a 3-step detergent-enzymatic extraction method involving the 1% deoxycholic acid (DOA) in combination with RNase (20μg/ml) and DNase (200μg/ml). HLA-DR antigen expression was detected by using immunohistochemical techniques. The valve and wall of the HV were transplanted subcutaneously in the mice for 8 weeks, and the histology, calcium assay and calcium content were examined. ResultsFor the staining of the HLA-DR antigens, the immunogenic potential of the HV with extracting all endothelial cells and interstitial cells or only the endothelial cells was lower than cryopreserved HV, but it more obviously decreased for the HV with extracting all endothelial cells and interstitial cells. After 8 weeks embedded in the mice, the histological signs of the inflammatory reactions and the calcification extent to the cryopreserved HV and the HV with only extracting endothelial cells were stronger than the HV with extracting all endothelial cells and interstitial cells predominantly. And calcification extent or the inflammatory reactions to the wall of the HV were more severe than those of the leaflet. ConclusionsThe immunogenicity of the HV with extracting all endothelial cells and interstitial cells is much less than HV with only extracting endothelial cells. The histological signs of the inflammatory reactions and the calcification extent in vivo experiments is obviously decreased. For the HV with only extracting endothelial cells, though the histological signs of the inflammatory reactions slightly decrease, the calcification extent in vivo experiments is more severe, especially for the wall. The interstitial cells may be the important factor for the donor-reactive immune responses that is related to the graft calcification or destruction after implantation.
With the development of social economy and medicine, degenerative heart valve disease has become the major part in heart valve disease. Calcific aortic valve disease (CAVD) is one of the most representative manifestations of degenerative valvular disease. Aortic valve calcification (AVC) has been found to be a strong predictor of major cardiovascular events, which makes it necessary to identify an effective way to evaluate the degree of AVC. Numerous methods of quantitative assessment of AVC have been reported. Here, we discuss these methods from the aspects of pathology and imageology.
Reports about the application of transcatheter aortic valve replacement (TAVR) for patients with aortic stenosis, whose valve sizes exceed the maximum recommended annular diameter of the largest artificial valve, is rarely in China. This paper reports an aortic stenosis patient characterized by large aortic annulus diameter with severe calcification and treated by TAVR. A comprehensive and careful operation plan was made before the operation. The anterior and posterior balloon dilatation and coronary artery protection were used during the operation. The patient was followed up for 2 years and was in stable condition.