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Reverse transcriptase-polymerase chain reaction (RT-PCR) for bone markers in diet, control, and reversibility group valves. RT-PCR for osteopontin, Runx-2, osteocalcin, and RANKL obtained from valves in all groups, demonstrating a significant up-regulation in expression of all four mRNA in the diet group valve tissue compared with control. There is downregulation of all four mRNA at the valves obtained from the reversibility subgroup.
