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Moreover, it has been suggested that coronary artery calcification may be one of the sequelae of severe vasculitis occurring in acute KD patients [ 27 , 28 , 29 ].
The clinical symptoms of vascular calcification in KD patients has already been demonstrated, however its mechanism is still unclear.
Although it still lacks an in vivo animal model or direct clinical evidence, the elucidation of the mechanism in this study could provide a new direction to explain the cause of KD vascular calcification.
VSMC calcification is one of the crucial components of cardiovascular disease [ 14 , 16 ]. Previous studies have demonstrated that both IP and IL can be secreted into the plasma and subsequently may play a role in the development of coronary arteritis in KD [ 7 , 8 , 9 , 10 , 11 , 12 , 13 ].
Inflammation is an important factor in the formation of atherosclerosis, as well as KD. It has also been indicated that the coronary artery calcification of KD appears to be initiated by intensive inflammatory stimulation [ 29 ].
Both IP and IL can be secreted from immune cells, i. Although still slightly controversial, recent studies have further indicated that IP and IL can be recognized as proinflammatory factors that may be involved in the development of coronary arterial disease, including aneurysms and atherosclerosis.
BMPs are context-dependent growth factors and thus possess different functional activities in different cell types.
In embryos, BMPs can control vascular angiogenesis and patterning [ 41 , 42 ]. In the present results, although higher levels of BMP6 were demonstrated in KD serum, we also found three samples from febrile patients that had similar results.
It has been suggested that more serum samples from KD and febrile patients are needed to be counted and studied. Moreover, both KD and febrile disease have been considered as a kind of inflammation.
Thus, our study also suggested that BMP6 may play a role in the inflammation regulation of KD and even febrile disease , however this role needs to be further examined in the future.
Additional in vivo experimental mouse models of KD and human studies will be needed to confirm the in vivo relevance of these in vitro findings. Therefore, this limitation will be addressed in our ongoing study and be further elucidated in future.
Overall, this in vitro study could provide new insights into the pathogenesis of vascular calcification in VSMCs in KD progression.
All others reagents and chemicals were obtained from Sigma St. Louis, MO. ARS an anthraquinone dye is a calcium-deposition detective reagent.
ARS stain has been widely used to analyze calcium-rich deposits in mineralizing tissues and cells for many decades [ 46 , 47 , 48 ].
After the treatment, the cells were fixed formaldehyde and then were stained with ARS reagent. These stained cells were further extracted by adding the acetic acid and then were neutralized by adding the ammonium hydroxide.
Cell lysates were collected by lysing cells with 0. The nitrocellulose paper was further hybridized and analyzed with the indicated antibodies and the detection system.
The clinical plasma samples from both the FC and KD patients were recruited and harvested. In all the assays, at least three individual experiments were carried out.
The data that support the findings of this study are available from the corresponding author upon reasonable request. Agarwal S, Agrawal DK.
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