Effect of Bushen Yiqi Hexue Recipe (补肾益气和血方) on Pinopodes Expression on Endometrial Surface in Embryo Implantation Dysfunctional Mice

Objective: To explore the therapeutic mechanism of Bushen Yiqi Hexue recipe (补肾益气血方, BYHR), and observe its effect on pinopodes expression on endometrial surface in mifepristone induced embryo implantation dysfunctional mice. Methods: Pregnant Kunming mice were randomly divided into the normal group, the control group and the treated group. Mice in the treated group were administered with BYHR, and those in the normal group and the control group were administered with normal saline, starting from the very first day of pregnancy (Pd1). On the Pd4, mifepristone (RU486) was subcutaneously injected into mice in the control group and the treated group. Pinopodes expression on endometrial surface at 09:30-10:00 pm on Pd4 (regarded as time point 1, T1) and at 09:30-10:00 am on Pd5 (as time point 2, T2) was determined by scanning electron microscopy. Pregnancy rate and average implanted embryos were observed on Pd7. Results: Pregnancy rate and average implanted embryos in the control group were obviously lower than those in the normal group (all P<0.01), while the two parameters were significantly higher in the treated group (P<0.05, P<0.01, respectively) than those in the control group. In the normal group, abundant developing pinopodes were distributed over the whole endometrial surface in T1, and they were altered to a great deal of fully developed pinopodes in T2. But in the control group, only a few pinopodes were expressed locally on endometrial surface in T1, showing a nonsynchronous figure of development, and they completely disappeared in T2. In the treated group, a lot of developing pinopodes were expressed like those in the normal group but somewhat lagged behind, whereas many fully developed pinopodes were expressed in T2. Conclusion: The decrease of the pinopodes in T1 and the cleaning up of them in T2 is possibly the partial mechanism of mifepristone in inducing embryo implantation dysfunction. It is indicated that BYHR could improve pinopodes expression on endometrial surface, eventually better the uterine receptivity and improve the embryo implantation.

Hysteroscopic Appearence of Midsecretory Endometrium in Relation to Pinopodes Expression and the Reproductive Outcome in Infertile Women

Objective To investigate the relation between hysteroscopic finding and pinopodes expression, meanwhile to explore whether hysteroscopic finding was a predictor of reproductive outcome for infertile women. Methods A total of 79 infertile patients underwent hysteroscopic assessement of the midsecretory endometrium and hormonal measurement 7 9 d after a spontaneous ovulation assessed with transvaginal ultrasound. Pinopodes expression by scanning electron microscope was examined simultaneously in randomly selected 32 patients. All patients were followed up to determine their reproductive outcomes. Results Twenty-six （32.9%） patients were diagnosed as having ＇good＇ midsecretory endometrium and 53 （67.1%） as ＇poor＇ midsecretory endometrium by hysteroscopy. There were no significant differences of endometrial thickness and pattern in the ＇good＇ group compared with the ＇poor＇ group. Serum progesterone level in the ＂good＇ group was apparently higher than that in the ＇poor＇ group （P=0.025）, but no correlation was observed between serum oestradiol level and hysteroscopic finding. Significant differences of the development stage and number of pinopodes were observed between the two groups （P〈O.05）. The pregnancy rate was significantly higher in the ＇good＇ group （53.9%） than in the ＇poor＇ group （26.4%） （P=O.O17）. Conclusion The hysteroscopic appearance of the midsecretory endometrium is associated with serum progesterone level and pinopodes expression, which provides prognostic information with regard to the reproductive outcome of infertile women.

子宫内膜轻创对着床相关因子及IVF妊娠结局影响的研究

目的:探讨子宫内膜轻创术对提高子宫内膜容受性的价值及分子机理。方法:对120例不孕症患者行IVF140周期,随机分为行子宫内膜搔刮轻创术组60例(A组)及对照组60例(B组),采用内膜微活检的方法,在扫描电镜下观察胞饮突,免疫组化定位及半定量检测白血病抑制因子(LIF)、骨桥蛋白(OPN)。再将对照组未孕患者中的20例行轻创术(B2组),重复IVF,并进行相关因子测定。结果:A组和B组相关因子分别为:胞饮突丰富占63.33%和36.67%、LIF6.07±1.33和4.94±2.26、OPN6.36±1.49和5.28±2.06、单胚种植率27.8%和14.3%,组间比较差异均有统计学意义(P<0.05)。B2组行轻创术前后比,分别为:胞饮突丰富10%vs50%,LIF4.50±0.82vs6.33±1.25,OPN5.06±0.98vs6.83±1.46,轻创术前后差异均有统计学意义(P<0.05)。按妊娠与否分组后,妊娠组(n=62)与非妊娠组(n=78)相关因子分别为:胞饮突丰富54.84%和25.64%,LIF6.94±1.03和4.24±1.84,OPN7.31±0.85和4.52±1.65,差异有统计学意义(P<0.05),且LIF与OPN间呈正相关。结论:胞饮突、LIF、OPN是子宫内膜着床期重要的相关因子,它们的高表达与子宫内膜容受性紧密相关。而轻创术可增强3者的表达,从而提高胚胎种植率,改善IVF妊娠结局。

输卵管积水妇女种植窗期子宫内膜胞饮小泡及部分种植因子的变化

目的:探讨输卵管积水对种植窗期子宫内膜胞饮小泡及种植因子integrinβ3、MUC1及LIF表达的影响。方法:选择接受IVF治疗的20例输卵管积水妇女(积水组)及21例因男性因素所致不孕的妇女(对照组),在种植窗期间通过扫描电镜观察子宫内膜表面胞饮小泡的形态、密度,免疫组织化学分析种植因子integrinβ3、MUC1及LIF的表达。结果:对照组成熟期胞饮小泡所占比例以及胞饮小泡的密度与积水组相比无统计学差异(P>0.05)。integrinβ3、LIF及MUC1在正常对照组的腺上皮细胞和腔上皮细胞中的表达强度均明显高于积水组,integrinβ3在间质细胞中表达亦明显高于积水组。结论:integrinβ3、MUC1及LIF受积水的影响较胞饮小泡更为敏感,先于子宫内膜表面超微结构受到改变,可能是造成胚胎种植率下降的主要原因之一。

Effects of Bu-Shen-An-Tai Recipe and Its Two Components on Endometrial Morphology during Peri-implantation in Superovulated Mice

The aim of this study was to investigate the effect of Bu-Shen-An-Tai recipe （BSATR） and its two components （Bushen recipe, and Huoxue recipe） on endometrial morphology during peri-implantation in superovulated mice. Mice were randomly divided into five groups, including the normal （N）, model （M）, Bushen （BS）, Huoxue （HX） and Bu-Shen-An-Tai （BH） groups. The uteri were collected on day 4 of pregnancy, and the endometrium thickness, microvessel density （MVD） and number of pinopodes observed. Compared with the M group, the endometrial thickness in the BS, HX and BH groups was significantly increased and there was a significant difference in endometrial thickness between the BS and the BH groups. The mean MVD was significantly lower in the M group than in the N group, and there was a significant increase in MVD in the BS, HX and BH groups as compared with the M group. Compared with the M group, the pinopode scores in the en- dometrium were significantly increased in the HX and BH groups; and the BS group had significantly higher pinipode scores than the HX and BH groups. In conclusion, the results of the present study demonstrated that the recipes （Bushen, Huoxue and BSATR） could improve the endometrial envi- ronment by regulating the endometrial thickness, MVD and the number of pinopodes at the window of implantation. Moreover, the Huoxue recipe and the BSATR were more efficient than the Bushen recipe, with the BSATR tending to have the most beneficial effects.

Endometrial Mucin-1 and Pinopode in Peri-implantation Phase in Ovarian High Responders during Controlled Ovarian Hyperstimulation Cycles

Objective To investigate effects of ovarian high response on endometrial mucin-1 （MUC1） and pinopode in peri-implantation phase in controlled ovarian hyperstimulation （COH） cycles. Methods Ovarian high response was defined as serum E2 〉 15 000 pmol/L on the day of hCG administration in COH cycle using GnRH agonist and recombinant FSH （n=8）. Healthy and fertile women were used as the natural control （n=10）. Endometrial biopsies were performed on the day of LH＋ 7/hCG＋ 7. Pinopode formation was observed by scanning electron microscope. Expression of MUC1 was detected with quantitative Real-time PCR and immunohistochemistry. Results In high response group, the lumen surface was covered with variant pinopodes and microvillous. The expression of MUC1 mRNA in high response group was lower than that in the natural control （P〈0.05）. Immunostaining for MUC1 protein in glandular and luminal epithelium in high response group was lower than that in the natural control （P〈0. 05）. Conclusion Asynchronized pinopode appearance and lower expression of MUC1 during peri-implantation period were the characteristics of endometrium in high response group, Which may provide a clue of decreased endometrial receptivity in the supraphysiological hormone milieu.