| Objectives 1.Evaluating the impacts of severe EMS on assisted reproductive outcomes,and analyzing the main influencing factors,which provides research direction for EMS-related infertility.2.Screening for proteins which are specifically expressed in the follicular fluid of infertile patients with severe ems and studying their impacts on oocytes quality.3.Exploring the mechanism behind the treatment of Neiyi Fang to EMS-related infertility from the research direction of oocyte quality.Methods 1.Clinical research By a retrospective cohort study with large sample,the characteristics and various indicators in oocytes retrieval cycles and transfer cycles of oviduct infertility patients(control group)and infertile patients with different types of severe EMS,were collected and analysed for the purpose of evaluating the influence of severe EMS on pregnancy outcomes in IVF/ICSI-FET and analyzing the main factors.2.Screening and verification of the proteins expressed specifically in follicular fluid of infertile patients with severe EMS Detect the follicular fluid of infertile patients with tubal factors and infertile patients with severe EMS to screen out the proteins with abnormal expression levels by i TRAQ.Select the proteins which are related with EMS and verify their differences in expression levels by ELISA with a larger sample.3.Based on the experimental results of “2”,detect the ferri ion level of the follicular fluid of infertile patients with severe EMS “2.Experimental results” showed that transferrin(TRF)was poorly expressed in the follicular fluid of infertile patients with severe EMS,and the protein was closely related to ferri transport.Ferri ion level in follicular fluid of infertile patients with severe EMS and infertile patients with tubal factors were detected by ELISA,and TRF saturation was compared between the two groups.4.The impact of the change of TRF level in follicular fluid of infertile patients with severe EMS on mouse oocyte quality Intervene in the maturation and fertilization in vitro of mouse oocytes and the early embryonic development with the follicular fluid of infertile patients with severe EMS,which were added TRF to the level of those of infertile patients with tubal factors or added TRF antibody to block TRF molecular function.5.Effect of Neiyi Fang on oocyte quality of the EMS model mouse EMS model mice were constructed and treated with Neiyi Fang.The natural fertility,follicular development,oocyte maturation in vitro and TRF expression in follicle was studied.Results 1.The level of serum FSH and E2 on the third day of menstruation in EMS group was significantly higher than that in the control group,while the antral follicle count was significantly lower than that in the control group.The antral follicle count in the ovarian endometrioma group was significantly lower than that in the pelvic EMS group.In terms of the number of dominant follicles,the number of retrieved oocytes,the number of mature oocytes,the rate of retrieved oocytes,the rate available blastocysts,the rate of high-quality embryos and the rate of transplantable embryos,the EMS group was significantly lower than the control group,but the incidence of poor ovarian response was significantly higher than the control group.In terms of the number of dominant follicles,the number of retrieved oocytes,the number of mature oocytes and the rate of retrieved oocytes,the ovarian endometriomas group was significantly lower than the pelvic EMS group,and the rate of fertilization and cleavage was significantly higher than the pelvic EMS group.The number of dominant follicles in operation group was significantly lower than that in puncture group.The abortion rate and cumulative clinical pregnancy rate was significantly lower in EMS group than those of control group.2.Isobaric tags for relative and absolute quantitation results showed that TRF level in the follicular fluid of the infertile patients with severe endometriosis(EMS group)was significantly lower than that of the infertile patients with tubal factors(control group)(P=0.043).The results of enzyme-linked immunosorbent assay showed that TRF concentration in EMS group(3.43±0.53 g/L)was significantly lower than that in control group(4.17±0.54 g/L)(P=0.042).3.The level of ferri ion in the follicular fluid of EMS group(40.14±2.34 umol/L)was significantly higher than that of control group(29.46±1.77 umol/L)(P<0.001),and the TRF saturation of EMS group was 54.8%,indicating iron overload.4.The in vitro maturation rate of oocytes in EMS group was 46.7%,significantly lower than that of control group(61.2%,P = 0.017)and the TRF group(65.7%,P = 0.001).The in vitro maturation rate of oocytes in the antibody group was 47.1%,significantly lower than that of the TRF group(P = 0.007).5.The mean number of birth of the Neiyi Fang group was significantly higher than that of the other three groups(all P < 0.001)in the copulation experiment.There was a statistically significant difference in the proportion of follicles in each stage between the Neiyi Fang group and EMS group(P=0.019),and the percentage of lumen follicles(late stage)in the EMS group(12.3%)was higher than that in EMS group(1.9%).The in vitro maturation rate of oocytes(68.3%)in Neiyi Fang group was significantly higher than that in EMS group(51.0%)(P =0.014).Conclusions 1.Severe EMS has adverse effects on the outcomes of IVF/ ICSI-FET,which is mainly reflected in the lower probability of clinical pregnancy(cumulative clinical pregnancy rate)after a single oocytes retrieval.One of the important reasons is the poor ovarian response and the decreased number of oocytes retrieved,especially in patients with ovarian endometriomas.EMS has other adverse impacts outside the ovary on the quality of oocytes,resulting in reduced capacity of fertilization and the number of fertilized oocytes which can cleave.The surgery of ovarian endometriomas can not improve pregnancy outcomes of IVF/ ICSI-FET.2.Transferrin level decrease and ferri ion level increase in the follicular fluid of infertile patients with severe EMS,in which iron overload occurs.3.The decrease of transferrin level is one of the reasons that the follicular fluid of infertile patients with severe EMS prevents in vitro maturation of mouse oocytes.4.Neiyi Fang can improve the natural fertility,promote follicular development and in vitro maturation of oocytes of EMS model mouse,indicating that it has improved effect on the quality of oocytes in EMS. |
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