The Analysis Of The Relevant Factors Affecting Pregnancy In Endometriosis And The Establishment Of Animal Models

1 Objective Exploring the effect of endometriosis(EMS)on the outcome of pregnancy after in vitro fertilization and embryo transfer(IVF-ET)and analyzing the factors related to pregnancy.Taking the mouse as the experimental object,to explore the most effective modeling method of animal model and time scales in the best state,to provide reference for future research,and prepare for the influence of endometriosis on mouse embryos.2 Method2.1 Clinical research55 oocyte retrieval cycles of 46 cases of infertility patients associated with endometriosis treatmented in our center between January 2012 and June 2016 were selected as endometriosis group(EMS group);And 156 oocyte retrieval cycles of 126 cases of non endometriosis patients during the same period were selected as control group(non EMS group).After balancing the general situation of the two groups,weanalyzed the flowing datasof the two groups retrospectively,which including CA-125 level,antral follicle counts(AFC),starting dose and total dose of ovulation induction drugs,thickness and types of endometrium on the day of injecting HCG(or the HCG day),number of mature follicles(diameter ≥ 14mm)on the HCG day,level of E2 on the HCG day,level of E2 per mature follicle on the HCG day, number of retrieved oocytes,fertilization types,number of MII,number of normal fertilization / cleavage,number of high quality embryo,normal fertilization / cleavage rate,high quality embryo rate,implantation rate,biochemical pregnancy rate,clinical pregnancy rate,early abortion rate,live birth rate andcancelrateof thecycle,in order to investigate the effect of EMS on laboratory and clinical outcomes in infertility patients with EMS.Then the clinical data of the two groups were divided into pregnancy group and non pregnancy group according to clinical outcomes,and the factors related to pregnancy outcome in EMS group and non EMS group were further clarified.2.2 Experimental research1)A total of 33 female KM mice with special pathogen free(SPF)at the age of6-week-old were selected and divided into three groups randomly,with 11 mice in each group.2)After one week for adapting to the new environment,we removal the leftuterine horn of the mice by surgical,and divide it into 4 sections,and cut the uterine cavity.Eventually we can get four uterus tissues in each mouse.Two pieces of the tissueswere sutured on abdominal wallsof both sides of incision in the direction of.endometrium face to abdominal wall.A tissue was cut into pieces and injected into the abdominal cavity of mouse.The remaining piece of tissue was sent for pathological examinationto confirm the tissue from the uterus of mouse.Three groups of mice were established endometriosis modelsall by this method.3)At the end of 2 weeks,3 weeks and 4 weeks after surgery,the three groups of mice were sacrificed respectively.Then we observed the morphology of ectopic focus of uterus tissues and statistical calculation was followed.(1)The survival rate of the mice,the formation rate of the lesionof different parts,the formation rate of cysts in ectopic foci and the success rate of the model were compared.(2)The success rates of the tissue fragments planting in the abdominal wall,omentum and intestinal wall werecompared in each group.(3)The success rate of the three modeling methods were compared in each group.The three methods were suture,fragment planting and suture+ fragment planting.(4)The mean volume of ectopic lesions in three groups was compared.3 Result3.1 Clinical research The levels of CA-125 and the canceling rate of the cycle in EMS group was significantly higher than the control group(P < 0.05);AFC,number of mature follicles on the HCG day,retrieved oocytes,MII,2PN,normal cleavage,high quality embryo and high quality embryo rate in EMS group were significantly lower than the control group(P < 0.05).The correlation regression coefficient between number of mature follicles on HCG day andinfertility outcomes in patients with EMS was less than 0(P< 0.05).3.2 Experimental research33 KM mice were selected to make EMS model in this study and 28 survived.(1)The estrous cycle did not distinguish before operation inall the three groups of mice,but there were no significant differences in the postoperative survival rate,the lesions formation rate,the vesicle formation rate and the success model rate among the three groups.(P > 0.05).(2)There was no significant difference among the three groups in the success rate of implantation of endometrial fragments on the abdominal wall,omentum and intestinal wall(P > 0.05).(3)Comparison of success rates of three ways of modeling including suture,fragment planting,suture + fragment planting in each group of three groups.Only the success rate of modelby plantinglower than the success rate ofmodelsby suturing and suture + fragment planting in the group of 4 weeks after operation,the difference was statistically significant(P < 0.05).The success rate of the latter two methods are 100%.(4)The average volume of ectopic lesions in the three groups of mice showed that the volume of ectopic lesions in the 2 week after operation was less than that of the mice in the 4 week after operation(4.00±2.85mm3 Vs.25.64±15.93mm3,P=0.001),the difference was statistically significant(P<0.05).4 Conclusion4.1 Clinical research Ovarian reserve function in infertility patients with endometriosis is decreased,and the quality of eggs and embryos are decreased,which is more likely to cancel cycles.But the egg maturation rate,normal fertilization rate and clinical outcome in infertile patients with endometriosis have no significant difference when compared with infertility patients without endometriosis.The number of mature follicles on HCG day is the protective factor of pregnancy in EMS patients.4.2 Experimental research1)2 weeks after surgery,the EMS model was successful,and there was no need for a unified estrous cycle before surgery.2)There was no difference in theimplantation rate of uterine tissues in the abdominal wall,omentum and intestinal wall.3)If the study ’s next step begins in the following 2-3 weeks after surgery,abdominal implantation of uterine tissue fragments can be used to establish the animal model;If the study ’s next step begins in the 4th week after surgery,suture on abdominal wall can be the best way.4)4 weeks after surgery,the average volume of ectopic foci was the largest,and the EMS model was the best.