| Early embryo death and miscarriages are unsolved problems in reproductive biology. According to references, 40%～60% embryos in mammal can not carry a pregnancy to term, 60% of which occurred before or after implantation stages. A substantial proportion of early embryo death and miscarriages are caused by chromosomal, anatomical and endocrinological abnormalities, up to 40% of fetal losses remain a consequence of unexplained aetiology. Recently, it is believed that unexplained fetal losses are related with disorder of cytokine net- works in fetomaternal interface. This paper took Toosendanin as a toxic agent, investigating embryotoxicity of Toosendanin on KM mice and immunological toxicological mechanism in uteris, finding out the relationships between embryotoxicity and immunological toxicological mechanism of Toosendanin.1. Purification of Toosendanin was performed by re-crystallizing technique with ethanol.Using thin-layer chromatology and high performance liquid chromatography(HPLC)to determine the content of Toosendanin and the results is 99.64%. It indicated that re-crystallizing technique is reliable and feasible in determining the content of Toosendanin.2. Taking 10% Propylene Glycol physiological saline as dissolvent for Toosendanin.20 of 90 mice which half is male and another half is female was used to measure 100% lethal dose first, the rest was divided into 7 groups randomly. The ratio of high dose group and low dose group is set up as 1.2. The LD50 values in this experiment is 13.84 mg/kg, with the 95% confidence limit of 13.84±1.45 mg/kg. 3. Virgin female mice weighing 18～25g were superovulated, and then mated with male mice. The acquired pregnant mice were divided into 6 groups for embryotoxicity research of toosendanin on fetuses of mice in 2-Cell, Morula, Blastocysts Stages. Control group was set up in each of these three stages. Every pregnant mouse of treated and control groups in the three stages was given the dose of 1/30 LD50 toosendanin and same amount of PBS 2 times by ip injection respectively. Superovulation need not be carried out for embryotoxicity study of toosendanin in Implantation Stage, but each pregnant mice in treated and control group was given the dose of 1/30 LD50 toosendanin and PBS 3 times by ip injection respectively. Finally, the number and configuration of embryo in uteri of all groups were counted and examined. Results: Total embryo of treated group In 2-Cell, Morula and Blastocysts Stages had no significance difference comparing with corresponding control group(P > 0.05); Number of normal and abnormal embryo of treated group in 2-Cell Stages had no significance difference comparing with control group either(P > 0.05); Number of normal and abnormal embryo of treated group In Morula and Blastocysts Stages had great significance difference comparing with corresponding control group(P < 0.01). Due to injection to the treated group 3 times in Implantation Stage, almost all the fetuses in uteri had been melting, and counting of them could not be performed while healthy fetuses could be seen and counted clearly in control group. There are only mild histological changes in maternal heart, liver, lung, kidney and uteri in the all treated groups. All above means that injecting ip a small amount of toosendanin to pregnant KM mice may has special embryotoxicity.4. Investigating the relationship between early embryo death inducing by Toosendanin and the numbers of CD4+,CD8+ T lymphocytes in maternal uteris.Using Immunhisto- chemistry technique to determine the CD4+,CD8+ cells, and results show that the numbers of CD4+,CD8+ cells in treated groups are all higher than in controlled groups in above three stages; The numbers of CD4+ lymphocytes in controlled groups ascend gradually, and in treated groups rise sharply first, then descend steeply; The numbers of CD8+ lymphocytes in controlled groups ascend gradually first, then descend steeply, and in treated groups descend gradually.These results indicated that enhancement in CD4+,CD8+ T lymphocytes in the uteris is...
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