This study was conducted to investigate the effect of PGI2 on early embryo development of sheep in vitro lacking of endogenous prostaglandins (PGs).The specific inhibitor NS398 and SC560 (specifically inhibiting the activity of COX-1 and/or COX-2) were added into the culture medium respectively to study the effect of COX-1 and COX-2 on embryonic development of sheep.Different concentrations of Iioprost were added respectively into the culture medium to investigate the effect of PGI2 on early embryo development of sheep in vitro.The results showed that there were no significant differences on the cleavage and blastocyst rates between the group only with NS398 and that with both NS398 and SC560 (P>0.05).There were significant differences between the group only with NS398 and control group (P<0.05).After adding NS398,different concentrations of exogenous Iloprost could replace the role of embryonic-derived PGI2,which substantially eliminated the adverse effects of COX-2 inhibitor on early embryo development in vitro.The appropriate concentration of Iloprost was 1×10-6 mol/L.H33342 staining showed no significant difference (P>0.05).The results demonstrated that embryonic endogenous COX-2 played a major role in regulating synthesis of PGI2 in early embryo of sheep.The exogenous Iloprost was able to compensate for the loss of endogenous PGI2 in embryos and remove the inhibition of NS398 on COX-2,ultimately promoting the development of early sheep embryos in vitro.

旨在研究外源添加PGE2(前列腺素E2)对缺少内源性前列腺素(PGs,Prostaglandins)的牛IVF早期胚胎外发育的影响.在牛IVF胚胎中抑制内源前列腺素合成再外源添加PGE2,以观察PGE2对牛IVF胚胎发育的作用以及起作用的胚胎阶段.并检测PGE2受体的mRNA水平.抑制胚胎内源性前列腺素合成限速酶COX-1和COX-2会使牛IVF胚胎卵裂率、8-16细胞率、囊胚率以及孵化率显著低于常规培养组;而外源添加PGE2能够补偿IVF胚胎因缺少内源性PGs导致的损伤,基本抵消COX-1和COX-2的特异性抑制剂对牛早期胚胎体外发育产生的不利影响;PGE2的添加浓度以1×10-8 mol/L为宜,并且其补偿作用发生在8细胞阶段之前.利用半定量RT-PCR未能检测到以上6个阶段IVF胚胎上EP1、EP2、EP3和EP4四种受体的mRNA.PGE2可以恢复缺少内源性PG的牛体外IVF早期胚胎的正常发育.

Objective To investigate the distribution of progesterone receptors(PR),induced by estrogen or progesterone,on the endometrium in between mice treated by estrus synchronization and those in natural estrus.Methods Totally 45 mice were divided into five groups: Group A: pseudopregnant group in natural estrus,Group B: pseudopregnant group in estrus synchronization,Group C: ovariectomy group,Group D: group treated by progesterone,and Group E: group treated by estrogen.On day 4,6,and 8 after mating,the murine uteri were obtained to observe and analyze the distribution of PR in endometrium by immunohistochemistry.Results The progesterone receptors distributed in the three types of endometrium cells among all the groups.The positively stained cell number of endometrium in the group of natural estrus was significantly different from that in the groups by estrus synchronization(P0.05).On day 4 and day 6,the positively stained cell number of all three cellular types in the group treated by progesterone was much higher than that in ovariectomy group(P0.05).In addition,the positively stained cell number in glandular epithelium and endometrial stroma of the estrogen-treated group was much higher than that in the ovariectomy group on day 4,day 6 and day 8(P0.05),but the positively stained cell number in luminal epithelium was much lower than that in the ovariectomy group(P0.05).Conclusion The PR distribution in mouse endometrium by estrus synchronization and that in natural estrus,is mainly induced by their endogenetic estrogen and progesterone.

Objective In this experiment,it is aimed to investigate the effect of feeding female mice with isoflavones on some physiological characteristics of their female offspring.Methods The ICR female mice were divided into the control(0 mg/kg),low soy isoflavone level(50 mg/kg) and high soy isoflavone level(400 mg/kg) diet groups separately at the gestation period and prior to conception.In addition,the state of vagina opening and vagina plug of offspring after treatment with soy isoflavones,the concentration of estrogen in their serum on Day 45 and Day 65 as well as their body,uterus and ovary weight,litter size and their birth weight,were also recorded and analyzed separately.Results The litter size of those female mice feeding with 50 mg/kg or 400 mg/kg soy isoflavones,was significantly higher than that in the control group(P0.05).The state of vagina opening and vagina plug of the offspring whose mothers feeding with 400 mg/kg soy isoflavones during their gestation period,was significantly delayed than that of control one(P0.05).Their body weight on Day45 and Day65,uterus weight on Day 65,the concentration of serum estradiol on Day 65 were all significantly lower than those in control group respectively,but the ovary weight on Day 65 was significantly higher than it(P0.05).The female F1 generation's litter size and concentration of serum estradiol on Day 45 were obviously affected due to their mothers intaking 50 mg/kg soy isoflavones during their gestation period,but there were no significant differences among those different groups of female F1 generation whose mothers intaking soy isoflavones between their weaning and pre-gestation period(P0.05).Conclusion Collectively,it can be concluded that the female F1 generation's some reproductive characteristics such as the initial oestrus,development of reproductive organs and concentration of serum estradiol were affected significantly,due to their mothers intaking 400 mg/kg soy isoflavones during their gestation period,but were not affected anymore since their mothers intaking soy isoflavones only during their non-gestation period.

This study was carried out to compare, the different processing methods for X-bearing sperm screening, antibiotic type and density screening in sperm-talp screening the optimal number of embryos in general culture of drop (50 μL) and compare the effect of early embryonic development potential by using different individual Holstein bull′s X-bearing sperm. Results indicated that the X-bearing sperm treated by centrifugation can attain optimal effect. It was possible to cause pollution in vitro fertilization by using different bull′ s X-bearing sperm, and their embryonic development in vitro were also at a very low level. In vitro embryonic development by using X-bearing sperm in medium with 10 μL/mL Cefoperazone Sodium, was much better than others antibiotic (the blastocyst rate 16.9%); and reducing the number of embryo on the general culture drop could inhibit the pollution which caused by X-bearing sperm IVF embryo. The in vitro fertilization could be adaptable while the bovine X-bearing sperms were treated by direct centrifuge processing and adding with 10 μL/mL Cefoperazone Sodium, but the effect of such kind of fertilization in vitro was much lower than that of normal one.