A negative regulator of ENaC in mouse endometrial epithelium which is important for liquid microenvironment during periimplantation. The abnormal expression of CFTR and 
ENaCa might disrupt the cellular microenvironment that is harmful for embryo implantation or maintain of pregnancy; or might result from degeneration or necrosis of decidua as a consequence of miscarriage. Benzoylaconine However, the up-regulation of CFTR and downregulation of ENaC-a in mechanism of miscarriage remains unclear. CFTR and ENaC-a are ion channels that are co-expressed in a variety of cell types, including human endometrial and mouse endometrial cells. A previous study conducted in mice during preimplantation suggested that the CFTR protein was predominantly found in stromal cells, but not in epithelial cells, while the ENaC protein was predominantly localized to both luminal and glandular epithelial cells. Interestingly, in our study, we observed that the location of CFTR and ENaC-a protein expression in mouse uteri was not the same as that in human uteri. There are several reasons that might lead to these inconsistencies. Firstly, polyclonal antibodies used in the study might result in Diacerein non-specific expression; Secondly, the human decidua samples were obtained from cases of missed abortion and not fresh inevitable abortion; therefore, the tissue morphology might have been abnormal and luminal epithelia may not have been present. The exact mechanism associated with the spontaneous abortion in this model is unclear, and most studies have focused on abnormal immune reactions at the maternal-fetal interface. Several studies have demonstrated the predominance of Th2-type cytokines over Th1-type cytokines at the maternal-fetal interface in cases of successful pregnancy. However, in the CBA6DBA/2 model, increased expression of Th1-type cytokines such as interleukin -1, interferon -c, and tumor necrosis factor -a, and decreased expression of Th2-type cytokines such as transforming growth factor -b2, IL-4, and IL-10 are responsible for the spontaneous abortion. Interestingly, significantly higher IL-1b levels were found in the uterine fluid from women with repeated pregnancy failure after in vitro fertilization/embryo transfer. The interaction between ion channel and Th1-type cytokines is unclear. Some studies on respiratory and gastrointestinal system have suggested that IL-1b specifically upregulated CFTR gene expression in human intestinal T84 cells. The most frequently reported techniques used for DNMT3A mutations include direct sequencing, high-resolution melting analysis and next generation sequencing. Sanger sequencing is particularly well-established techniquefor the identification of previously unreported mutations but its relatively low sensitivitymay be problematic for detection of low frequency somatic mutations. High resolution meltingtechnique has also been adapted for detection of DNMT3A mutations and is also good for screening for unknown mutations in a single tube format with a sensitivity of about 4%. However, it requires well-established standards and eventually verification of the result through direct sequencing. Targeted amplicon resequencing on next generation sequencingplatforms has also been used for DNMT3A mutations. The obvious advantages of NGS as the digital allele burden output, theoretically very high sensitivity, the possibility for identification of novel mutations are currently limited by the costly equipment and the necessity for a strong bioinformatic support.