A graphical representation of all replicate primer-extensions normalized to 5S rRNA is plotted in Figure 2B. Upon Toxoplasma infection, miR-17 family members collectively increased,3-fold, and miR-19 and miR-25 family members increased,1.5-fold. When normalized to 5S rRNA, miR-18 also increased upon Toxoplasma infection, but the measurements for the miR-18 primer-extensions were complicated by low signal and high Paederoside background, making its measurement less reliable; it was therefore not included in Figure 2B. Similar primer-extension analyses were performed with probes for the miR-17 family and 5S rRNA on RNA samples derived from two Toxoplasma-Eupalinilide-C infected mouse embryonic fibroblast lines and from a human B-cell line ; the results of these experiments were consistent with the primary HFF results detailed above. The data from the miRNA microarray profiling and primerextension analysis were generally in agreement except in one instance. The microarray data indicated that the miR25 family of miRNAs did not appreciably increase in arrays hybridized with RNA derived from Toxoplasma-infected cells at the 24 hour timepoint whereas primer-extension for miR-92 indicated a modest but reproducible increase. The Cy5 channel intensities of the miR-25 family probe spots were near the upper limits of the dynamic range of the microarrays, which likely obscured the signal increase seen in the primer-extensions. Excluding this discrepancy, the primer-extension data closely approximate the results of the northern blot and miRNA microarray profiling and confirm that Toxoplasma increases the levels of mature miR-17, miR-18, miR-19 and miR-25 family members in infected human cells. Congenital heart disease is cardiovascular malformation caused by abnormal development of the fetal heart and the great vessels. Its incidence is 32.74/10000 in China but 5.4-16.1/ 1000 in the other countries. CHD is one of the most common types of congenital malformation in children, and is also one of the main causes of neonatal and infant mortality. The causes of CHD are complex, and are not fully understood. Most CHD may be related to genetic and environmental factors.