Aim： To investigate whether adriamycin induces DNA damage and the formation of γH2AX （the phosphorylated form of histone H2AX） foci in mature spermatozoa. Methods： Human spermatozoa were treated with adriamycin at different concentrations, γH2AX was analyzed by immunofluorescent staining and flow cytometry and doublestrand breaks （DSB） were detected by the comet assay. Results： The neutral comet assay revealed that the treatment with adriamycin at 2 μg/mL for different times （0.5, 2, 8 and 24 h）, or for 8 h at different concentrations （0,4, 2 and 10 μg/mL）, induced significant DSB in spermatozoa. Immunofluorent staining and flow cytometry showed that the expression of γH2AX was increased in a dose-dependent and time-dependant manner after the treatment of adriamycin. Adriamycin also induced the concurrent appearance of DNA maintenance/repair proteins RAD50 and 53BP 1 with γH2AX in spermatozoa. Wortmannin, an inhibitor of the phosphatidylinositol 3-kinase （PI3K） family, abolished the co-appearance of these two proteins with γH2AX. Conclusion： Human mature spermatozoa have the same response to DSB-induced H2AX phosphorylation and subsequent recruitment of DNA maintenance/ repair proteins as somatic cells.
The success of embryo implantation is a critical step towards further embryo development and pregnancy outcome. The observations and investigations on embryo implantation have been over a century. A huge body of knowledge has been accumulated in anatomy, histology, ultrastructure and hormonal regulation; as well as recently in depth information about molecular signaling pathways got from studies of genomic wide gene screening and specific gene deletion. The knowledge from basic research has also substantially helped to initiate and improve the Artificial Reproductive Technology (ART) in clinical applications. Now we've known that the normal embryo implantation involves the embryo's development into an implantation-competent blastocyst and the synchronized transformation of uteri into a receptive stage. The interdependent relationship between the blastocyst and uterus involves complicated hormonal regulation and local paracrine, juxtacrine interactions. In this paper, we review some important historical findings regarding uterine receptivity and blastocyst activation, as well as some less discussed topics such as embryo spacing, embryo orientation. Further understandings on detailed mechanisms during the process of embryo implantation will help cure women infertility as well as develop new generation of non-steroids contraceptives.
