Removal of hydrosalpinges increases endometrial leukaemia inhibitory factor (LIF) expression at the time of the implantation window

BACKGROUND: The presence of hydrosalpinges is associated with lower implantation and pregnancy rates in women undergoing IVF-embryo transfer, while salpingectomy improves these parameters. Although the mechanism by which hydrosalpinges affects fertility is not entirely understood, an adverse effect on endometrial receptivity has been postulated. In this study, we hypothesized that the adverse effects of hydrosalpinges on fertility may be in part mediated by inappropriate endometrial expression of leukaemia inhibitory factor (LIF), a cytokine implicated in implantation. METHODS: In order to test our hypothesis, we prospectively examined the expression of LIF during the window of implantation in the endometrium of infertile women (n = 10) with hydrosalpinges prior to and following salpingectomy and of fertile controls (n = 10) by Western blotting and immunohistochemistry. RESULTS: LIF expression was significantly lower in infertile women with hydrosalpinges compared with fertile controls (P < 0.05). Salpingectomy resulted in an increase in LIF expression in eight out of 10 women with hydrosalpinges. LIF levels were increased by 231 +/-49% (mean +/- SEM) following salpingectomy. Immunohistochemical analysis confirmed the Western blot findings. The increased LIF immunoreactivity was predominantly localized to luminal and glandular epithelial cells. CONCLUSIONS: Our findings suggest that observed benefit from salpingectomy in infertile women with hydrosalpinges may be in part mediated by the up-regulation of endometrial LIF expression.     

A prospective evaluation of the effect of salpingectomy on endometrial receptivity in cases of women with communicating hydrosalpinges.

BACKGROUND: We aimed to assess whether salpingectomy in women with communicating hydrosalpinges influenced endometrial receptivity. METHODS: The inclusion criteria were: women with communicating hydrosalpinges, absence of other confounding infertility factors and aged <40 years. Patients were scheduled for laparoscopy during the putative window of implantation (cycle days 19-21). In patients in whom salpingectomy was decided upon due to the severity of tubal disease (n = 10), an intra-operative endometrial biopsy was performed. Post-treatment endometrial sampling was done between day 19-21 of the fourth consecutive cycle. Pre-treatment and post-treatment samples were assessed by both conventional histologic criteria and alpha(v)beta3 integrin immunostaining, where histological score (HSCORE) was used for quantification. RESULTS: Despite normal histological maturation assessed by conventional criteria, 8/10 hydrosalpinx cases yielded an epithelial HSCORE of <0.7, which was below the accepted threshold. Following salpingectomy, luminal endometrial epithelium demonstrated a significantly increased alpha(v)beta3 integrin expression (Wilcoxon's signed rank test, P = 0.017). Although the mean HSCORE for glandular epithelia improved, it failed to reach statistical significance. Ultrasound visible hydrosalpinges (n = 5) and non-visible cases (n = 5) were also compared. However, neither the pre-treatment integrin expression, nor the postoperative improvement were significantly different between these groups. CONCLUSIONS: We conclude that the surgical treatment of communicating hydrosalpinges may improve endometrial receptivity as assessed by alpha(v)beta3 integrin expression. Women with hydrosalpinges may undergo endometrial evaluation by the molecular markers of implantation, such as alpha(v)beta3 integrin. This evaluation may be decisive in determining the optimal management of cases, and may also be used to assess the efficacy of the treatment. The expression of the implantation markers should be correlated with implantation and clinical pregnancy rates in IVF-embryo transfer programs.     

Hydrosalpinges adversely affect markers of endometrial receptivity

While in-vitro fertilization (IVF) was initially developed in women with tubal factor infertility, recent clinical studies have suggested that the presence of hydrosalpinges lowers implantation and pregnancy rates. We postulated that these hydrosalpinges cause impaired endometrial receptivity. A total of 103 women with hydrosalpinges were prospectively evaluated, and compared with 55 infertile and 44 fertile controls. All women had endometrial biopsies during the window of implantation, analysed by conventional histological criteria, and also stained for three integrin markers of endometrial receptivity (alpha1beta1, alpha4beta1 and alpha vbeta3). Women with hydrosalpinges (cases) expressed significantly less of the alpha vbeta3 integrin compared with controls. There was no difference in expression of alpha1beta1 or alpha4beta1 among groups. A significantly greater number of cases had out of phase histology and missing alpha vbeta3 (type I defects) and absent integrin expression despite normal histological maturation (type II) defects, compared with controls. Of 20 women with impaired endometrial receptivity who were also biopsied after hydrosalpinx surgery, 70% demonstrated increased alpha vbeta3 expression. Seventy-seven percent of type I and 57% of type II defects were corrected postoperatively. Using markers of endometrial receptivity, this study demonstrates that inflammatory hydrosalpinges have an adverse effect on endometrial receptivity, which in some cases may be overcome by surgical treatment of the hydrosalpinx.      

SHORT COMMUNICATION: Correlations between serum oestradiol level on day of HCG and speed of embryonic development in in-vitro fertilization

The number of embryos transferred, ease of embryo transfer,and rate of embryo cleavage are significant factors affectingthe success of IVF. High levels of oestradiol occurring whenHCG is given may reduce implantation and pregnancy rates. Adiscrepancy between the stage of embryonic development and thephase of endometrial maturation at embryo transfer may be responsiblefor IVF failure in some cases.     

The role of the endometrium and embryo in human implantation

Despite many advances in assisted reproductive technologies (ART), implantation rates are still low. The process of implantation requires a reciprocal interaction between blastocyst and endometrium, culminating in a small window of opportunity during which implantation can occur. This interaction involves the embryo, with its inherent molecular programme of cell growth and differentiation, and the temporal differentiation of endometrial cells to attain uterine receptivity. Implantation itself is governed by an array of endocrine, paracrine and autocrine modulators, of embryonic and maternal origin. Implantation failure is thought to occur as a consequence of impairment of embryo developmental potential and/or impairment of uterine receptivity and the embryo-uterine dialogue. Therefore a better comprehension of implantation, and the relative importance of the factors involved, is warranted. New techniques for monitoring changes in the endometrium and/or the embryo at the level of gene regulation and protein expression may lead to the identification of better markers for implantation. Moreover, the use of predictive sets of markers may prove to be more reliable than a single marker. Continuing refinements to ART protocols, such as optimizing ovarian stimulation regimens, the timing of human chorionic gonadotrophin injection, or the timing of embryo transfer, should help to increase implantation rates further.     

Endometrial pinopodes indicate a shift in the window of receptivity in IVF cycles

The formation of endometrial pinopodes detected by scanning electron microscopy may be a specific marker for uterine receptivity. Aiming to assess the effects of ovarian stimulation on pinopode formation, we examined sequential endometrial biopsies from 17 oocyte donors. Seven normally menstruating women served as controls. Up to four samples were taken from each woman at 24-72 h intervals between days 14 and 24, giving a total of 69 samples. The day of oocyte retrieval was designated day 14 in ovarian stimulation cycles and the day of luteinizing hormone surge was designated day 13 in natural cycles. Endometrial morphology and pinopode numbers were similar in both groups. Fully developed pinopodes appeared in only one sample per cycle, indicating their short life span. However, the cycle day these structures appeared varied up to 5 days between women and the distribution was as follows: day 18 (n = 2), day 19 (n = 7), day 20 (n = 4), day 21 (n = 3), day 22 (n = 1) in ovarian stimulation cycles, and day 20 (n = 2), day 21 (n = 2), day 22 (n = 3) in natural cycles. Furthermore, accelerated pinopode formation in ovarian stimulation cycles was positively correlated with day 13 progesterone. Our findings show that ovarian stimulation does not affect endometrial pinopode formation in terms of quantity and life span. The cycle days when pinopodes form are specific to the individual, being on average 1-2 days earlier in ovarian stimulation than in natural cycles. These changes in pinopode expression may reflect shifts in the window of receptivity, resulting in ovo-endometrial asynchrony and limiting implantation success in in-vitro fertilization.     

Ultrasound-guided hydrosalpinx aspiration during oocyte collection improves pregnancy outcome in IVF: a randomized controlled trial

BACKGROUND: Hydrosalpinges have adverse effects on IVF outcomes. Salpingectomy is effective in improving outcomes, but it is not always practical or safe. Ultrasound-guided aspiration of hydrosalpinges at oocyte collection is an option for those who develop hydrosalpinges during controlled ovarian stimulation; however, there is no randomized evidence to show whether this practice is effective. METHODS: Between October 1999 and June 2003, consenting women of age 相似文献   

Acquired and inherited thrombophilia: implication in recurrent IVF and embryo transfer failure

BACKGROUND: The objective of this study was to determine the incidence of undiagnosed thrombophilic factors and its relation to IVF and embryo transfer failure in women who have had three or more previous IVF-embryo transfer cycles. METHODS: The study group comprised of 90 consecutive women with three or more previously failed IVF-embryo transfer cycles (group A). Two control groups were enrolled: group B (n=90) included women who have had successful pregnancy after their first IVF-embryo transfer cycle, and group C (n=100) included women who conceived spontaneously with at least one uneventful pregnancy and no previous history of miscarriage. All women were tested for the presence of inherited [factor V Leiden (FVL) mutation, prothrombin mutation, methylenetetrahydrofolate reductase (MTHFR) mutation and deficiencies in proteins S and C and antithrombin III] or acquired (lupus anticoagulant and anticardiolipin) thrombophilic factors. RESULTS: An increase in the incidences of FVL, MTHFR and antiphospholipid antibodies was found in the study group compared with the two control groups. At least one inherited or acquired thrombophilic factor was detected in 68.9% of women with repeated IVF failure compared with 25.6 and 25% in the groups B and C, respectively (P<0.01). Combined thrombophilia (two or more thrombophilic factors) was significantly higher in women who have had repeated IVF failure as compared with the two control groups (35.6 versus 4.4 and 3%) (P<0.0001). CONCLUSION: Thrombophilia has a significant role in IVF-embryo transfer implantation failure. Women with repeated IVF-embryo transfer failure should be screened for thrombophilia.     

Endocrinology: Physiological oestradiol and progesterone replacement cycles in women with ovarian failure: a model to study endometrial maturation and sex steroid receptor regulation by exogenous hormones

High endometrial receptivity has been achieved with physiologicaloestradiol and progesterone replacement cycles in women withovarian failure. To understand whether different protocols usingthe oral route or the transdermal route can influence the endometrialmaturation and the regulation of sex steroid receptors, we studied33 women with ovarian failure treated by two commonly used protocolsand assessed endometrial receptivity using light microscopy,scanning electron microscopy and immunohistochemistry for oestrogenand progesterone receptors on biopsies taken to include differentperiods of the luteal phase. The morphology in these patientswas similar to that observed in women with normal ovulatorycycles, indicating that the morphological response is not dependenton the type of oestradiol, oral or transdermal, in the replacementcycles as compared to the endogenous oestradiol in the menstrualcycle. The relative distribution of steroid receptors betweenthe epithelium and stroma varies similarly to that observedduring the luteal phase of the menstrual cycle. These resultsconfirm the role of progesterone, especially the importanceof the number of days of exposure to it, in the disappearanceof steroid receptors from endometrial glands. These observationsgive a better understanding of endometrial receptivity aroundthe time of presumed implantation and confirm clinical resultsconcerning the best timing of oocyte transfer.     

Hormonal stimulation for IVF treatment positively affects the CD56bright/CD56dim NK cell ratio of the endometrium during the window of implantation

The effects of hormone stimulation for IVF treatment on endometrial receptivity remain controversial. Since CD56(bright) natural killer (NK) cells in the endometrium positively contribute to implantation and decidualization whereas CD56(dim) NK cells are negatively associated with reproduction, shifts in the balance between those cells will affect receptivity. Therefore, we compared the leukocyte composition in the endometrium of IVF women (n=20) with non-pregnant women (n=18) in a natural cycle, as a parameter for endometrial quality. Biopsies were obtained 7 days after ovulation. Histological dating of the endometrium showed no increased endometrial advancement after IVF treatment as compared to the control group. Flow cytometric analysis of leukocyte subsets showed that hormonal stimulation positively affected the CD56(bright)/CD56(dim) ratio in the endometrium by a relative decrease in the cytotoxic CD56(dim)CD16(+) NK cell numbers. The relative number of T-cells remained unaffected, while the number of non-T and non-NK cells (i.e. B-cells and macrophages) was higher in the IVF group. These effects were restricted to the endometrium and not observed in peripheral blood. Within the CD56(bright) population we could identify a distinct subset of NK cells (CD56(superbright)) that was unique for the endometrium. We conclude that hormonal stimulation for IVF treatment positively affects the CD56(bright)/CD56(dim) ratio of the endometrium during the window of implantation and does not affect endometrial advancement.     

Implantation: Low implantation rate after in-vitro fertilization in patients with hydrosalpinges diagnosed by ultrasonography

The purpose of this study was to assess whether patients withtubal infertility and a hydrosalpinx have a reduced implantationrate after in-vitro fertilization. The study included 741 patientswho had 1190 consecutive oocyte aspirations. The presence orabsence of hydrosalpinges was assessed by transvaginal ultrasonographyon day 2 of all cycles. In 62 patients treated in 104 cyclesa hydrosalpinx was diagnosed, whereas 493 patients treated in813 cycles had no hydrosalpinx and eight patients treated in16 cycles had uncertain hydrosalpinx. The results show thatthe presence of a hydrosalpinx is associated with a reducedpregnancy rate per aspiration (19.2 versus 32.6% P < 0.01),reduced implantation rate (2.9 versus 10.3%, P < 0.0005),reduced delivery rate per aspiration (5.8 versus 20.9%, P <0.0005), reduced delivery rate per embryo transfer (6.6 versus22.8%, P < 0.0005) and increased early pregnancy loss (70versus 36%, P < 0.005). Among 178 patients with unexplainedinfertility or other infertility factors treated with 257 aspirationsthe results were similar to those in patients with tubal infertilitywithout a hydrosalpinx. In conclusion, the presence of a hydrosalpinxdoes not impair the number of embryos transferred but seemsto impair the implantation process. We hypothesize that thismay be due to leakage of fluid into the uterine cavity whichmay disturb the receptivity of the endometrium and/or the developingembryos.     

Hydrosalpinx reduces in-vitro fertilization/embryo transfer pregnancy rates

A retrospective study was designed to examine whether the presenceof a hydrosalpinx influenced pregnancy outcome following in-vitrofertilization (IVF) treatment In stimulated cycles. A totalof 254 women with tubal lesions were included. Patients withone or two hydrosalpinges were compared with patients havingother tubal lesions and pregnancy outcome was analysed. Bothpregnancy and delivery rates were significantly higher in thepatient group without a hydrosalplnx. The results after frozenembryo transfer showed the same tendency. These findings stronglyindicate that a permanent hydrosalpinx has a negative influenceon implantation, as well as on pregnancy, and suggest that amore active approach against large hydrosalpinges should beundertaken before I.VF treatment, in order to improve the pregnancyrates.     

Variable aneuploidy mechanisms in embryos from couples with poor reproductive histories undergoing preimplantation genetic screening

BACKGROUND: Preimplantation genetic screening (PGS) is used to determine the chromosome status of human embryos from patients with advanced maternal age (AMA), recurrent miscarriage (RM) or repeated implantation failure (RIF). METHODS: Embryos from 47 such couples were investigated for chromosomes 13, 15, 16, 18, 21 and 22 using fluorescence in situ hybridization with two rounds of hybridization. The investigation included parental lymphocyte work-up, the screening of blastomeres on day 3 and full follow-up on day 5/6 of untransferred embryos. RESULTS: The outcome of 60 PGS cycles is described, in which 523 embryos were biopsied; 91% gave results, of which 18% were diploid for all the chromosomes tested and 82% were abnormal. The pregnancy rate per cycle that reached the biopsy stage was 27%, and 30% per embryo transfer. Satisfactory follow-up was obtained from 353 embryos; all those diagnosed as abnormal were confirmed as such, although two false-positives were detected in relation to specific chromosome abnormalities. Meiotic errors were identified in 16% of embryos. Between the RM, AMA and RIF groups, there was a significant difference in the distribution of embryos that were uniformly abnormal and of those with meiotic errors; with an almost 3-fold increase in meiotic errors in the first two groups compared with the RIF group. CONCLUSIONS: This complete investigation has identified significant differences between referral groups concerning the origin of aneuploidy in their embryos.     

Ovarian stimulation with GnRH agonist, but not GnRH antagonist, partially restores the expression of endometrial integrin beta3 and leukaemia-inhibitory factor and improves uterine receptivity in mice

BACKGROUND: The impact of different ovarian stimulation (OS) protocols on endometrial receptivity remains controversial. In this study, the effects of different OS on the expression of endometrial integrin beta3 subunit and leukaemia-inhibitory factor (LIF) during the implantation window and the implantation rate in mice were investigated. METHODS: Three OS protocols were used, involving either pregnant mare's serum gonadotrophin (PMSG) alone, PMSG plus GnRH agonist or PMSG plus GnRH antagonist. Uterus samples were collected at 48 h after OS or ovulation and were detected with immunohistochemistry, Western blot and RT-PCR analyses. Normal embryos at gestation day 4 were transferred into the uteri of mice in the control and OS groups. RESULTS: All OS groups showed a significant decrease in the expression of both the endometrial integrin beta3 subunit and LIF during the implantation window and the implantation rate. Among the three OS groups, GnRH agonist-treated mice showed a higher endometrial integrin beta3 subunit and LIF expression and a higher implantation rate. No significant difference was found in the measured indices between the GnRH antagonist and PMSG groups. CONCLUSIONS: OS may inhibit the expression of endometrial integrin beta3 subunit and LIF and impair endometrial receptivity in mice. OS with GnRH agonist, but not GnRH antagonist, may partially restore the endometrial physiological secretion and improve uterine receptivity.     

Ovarian stimulation for in vitro fertilization alters the protein profile expression in endometrial secretion

Failure of the embryo to implant now constitutes the major limiting step in IVF treatment. Successful implantation requires a vital embryo and an effective molecular dialogue with a ‘receptive’ endometrium. However, what precisely constitutes a receptive human endometrium remains poorly defined. Several observations have indicated that ovarian stimulation for IVF may impair endometrial receptivity. The histological approach to monitor endometrial maturation requires an invasive biopsy that excludes its use during the luteal phase of cycles in which implantation is the end-point objective as in IVF. In recent years, several studies have been reported that the removal of endometrial secretions immediately prior to embryo transfer provides sufficient material for analysis of markers of receptivity without disrupting embryo implantation. Therefore, analysis of protein patterns in endometrial secretion fluid may offer a relatively non-invasive means of assessing endometrial receptivity during fertility treatment cycles. Several studies have shown that protein profile expression in endometrial secretions undergo cyclical changes, and demonstrated significant differences between the natural cycle and stimulated cycle. These findings suggest that endometrial secretion analysis provide a novel means of investigating the effect of ovarian stimulation on the intrauterine environment at the time of embryo transfer, which may help to develop less disruptive ovarian stimulation protocols for IVF in the future.     

Nitric oxide donors for patients undergoing IVF. A prospective,double-blind,randomized, placebo-controlled trial

BACKGROUND: Nitric oxide (NO) is involved in local control of the uterine cycle and in preparation of the uterus for pregnancy. NO donors, acting as vasodilating agents, may therefore have possible therapeutic uses, for example they may be of benefit to patients with a history of implantation failure. In a prospective, comparative, randomized, placebo-controlled study, we assessed the efficacy of nitroglycerin (NTG) administered to 138 IVF patients with a history of implantation failure. METHODS: Controlled ovarian stimulation was performed with long agonist protocol combined with recombinant FSH. Embryos were transferred on day 2 or 3 after oocyte retrieval. Eligible patients were those who had at least two 'good quality' embryos. The NTG patch was administered the day before embryo transfer and continued until either the results of the pregnancy test were known or until menstruation occurred. RESULTS: Ovarian response, implantation rate and pregnancy rate were comparable between both groups. No difference was observed in uterine Doppler findings, particularly the mean pulsatility index. CONCLUSIONS: NTG treatment the day before embryo transfer was no more effective than placebo in improving the implantation or pregnancy rates in a population of IVF patients with a previous history of implantation failures.     

The endometrium in stimulated cycles for IVF

Ovarian stimulation for IVF is known to affect luteal phase function. The endometrium in IVF cycles is thus subject to an altered endocrinological environment and to a possible direct effect of the ovarian stimulation therapy. Factors influencing the endometrial receptivity in such cycles are poorly understood. Studies comparing the endometrium in IVF cycles with natural cycles as controls have shown premature secretory changes in the post-ovulatory and early luteal phase of IVF cycles, followed by a large proportion of dyssynchronous glandular and stromal differentiation in the mid-luteal phase. These findings suggest a profound modification of luteal endometrial development in stimulated cycles. This hypothesis is further supported by the demonstration of a modified endometrial steroid receptor regulation and a profound antiproliferative effect in IVF cycles. The time of maximal endometrial receptivity is defined as the implantation window and is characterized by the expression of various endometrial products, among which pinopodes, integrins and leukaemia inhibitory factor are best described. Premature expression of pinopodes and integrins are in line with the observation of precocious luteal transformation following ovarian stimulation, although the clinical relevance with respect to the establishment of a clinical pregnancy awaits further validation. Studies exploring the endometrium within the cycle of embryo transfer have shown a deleterious effect of severe peri-ovulatory maturation advancement exceeding 3 days, as no clinical pregnancies were obtained in this condition. Further unravelling of molecules involved in the implantation mechanism is needed for a better comprehension of the link between altered endometrial development and receptivity in IVF cycles.     

多组学技术在人子宫内膜容受性中的研究进展

子宫内膜容受性是胚胎成功着床的关键,其异常是胚胎着床失败及不孕的主要原因,而传统的子宫内膜形态学观察方法无法充分确定子宫内膜容受性的状态。近年来,多组学技术的发展为生理病理条件下的子宫内膜功能特征研究提供了便捷有效的分析方法。因此,本文对转录组学、蛋白质组学及代谢组学等在人类子宫内膜容受性相关研究中的应用进行综述,旨在深入了解子宫内膜容受性并发掘新技术以强化子宫内膜植入窗口期检测的准确性,从而提高辅助生殖技术周期中怀孕的几率。     

Hydrosalpinx affects the implantation of previously cryopreserved embryos

The presence of hydrosalpinx has been reported to negativelyaffect the pregnancy and implantation rate after in-vitro fertilization(IVF) with embryo transfer. Hydro-salpinges are able to enlargeduring ovarian stimulation with a possible increased passageof tubal fluid into the endometrial cavity. We report the effectof hydrosalpinges during the transfer of previously cryopreserved/thawedembryos during a natural cycle. In all, 14 transfers in 10 patientswith a sonographically-documented hydrosalpinx during the studiedcycle (group I) were compared to 98 cycles in 74 patients withtubal disease but no such sonographic finding (group II). Bothpregnancy and implantation rates were significantly lower ingroup I (7.14 versus 24.49% and 5.0 versus 10.8% respectively).The presence of hydrosalpinx negatively affects pregnancy andimplantation rates during natural cycles.