Role of osteopontin in decidualization and pregnancy success.

OPN is essential for blastocyst implantation and placentation. Previous study found that miR181a was increased while miR181b was downregulated in endometrium during decidualization. However, the information regarding their effects on decidualization in human endometrium is still limited. Here, we report a novel role of OPN and miR181b in uterine decidualization and pregnancy success in humans. The expression of OPN was high in endometrium in secretory phase and in vitro decidualized hESC, whereas miR181b expression was low in identical conditions. Further analysis confirmed that OPN expression was upregulated by cAMP and C/EBPß signal pathway, while downregulated by miR181b. Increased OPN expression could promote the expression of decidualization-related and angiogenesis-related genes. Conversely, the processes of decidualization and angiogenesis in hESC were compromised by inhibiting OPN expression in vitro OPN expression was repressed in implantation failure group when compared with successful pregnancy group in IVF/ICSI-ET cycles. These findings add a new line of evidence supporting the fact that OPN is involved in decidualization and pregnancy success.

Osteopontin is expressed in the oviduct and promotes fertilization and preimplantation embryo development of mouse.

Osteopontin (OPN) is a multifunctional phosphoprotein that is detected in various tissues, including male and female reproductive tracts. In this study, we evaluated OPN expression in mouse oviducts during the estrus cycle, and at days 1-5 of pregnancy and pseudopregnancy by reverse transcription polymerase chain reaction (RT-PCR) and immunohistochemistry. The mice oocytes, sperm and embryos were treated with different concentrations of anti-OPN antibody in vitro to detect the function of OPN in fertilization and preimplantation embryo development. OPN mRNA and protein expression in mouse oviducts were cyclic dependent throughout the estrous cycle, which was highest at estrous and lowest at diestrous. Such a phenomenon was consistent with the change in estrogen level in mice. The expression levels of OPN in mice oviduct of normal pregnancy and pseudopregnancy were significantly different, which indicated that OPN expression in mouse oviducts was depend on estrogen and preimplantation embryo. Furthermore, anti-OPN antibody treatment could reduce the rates of fertilization, cleavage and blastocyst formation in vitro in a dose-dependent way. Overall, our results indicated that the expression of OPN in mouse oviducts during the estrous cycle and early pregnancy is likely regulated by estrogen and the embryo, and OPN may play a vital role in oocyte fertilization and preimplantation embryo development.

Uterine micro-environment and estrogen-dependent regulation of osteopontin expression in mouse blastocyst.

Embryo implantation is a highly synchronized bioprocess between an activated blastocyst and a receptive uterus. In mice, successful implantation relies on the dynamic interplay of estrogen and progesterone; however, the key mediators downstream of these hormones that act on blastocyst competency and endometrium receptivity acquisition are largely unknown. In this study, we showed that the expression of osteopontin (OPN) in mouse blastocysts is regulated by ovarian estrogen and uterine micro-environment. OPN mRNA is up-regulated in mouse blastocyst on day 4 of pregnancy, which is associated with ovarian estrogen secretion peak. Hormone treatment in vivo demonstrated that OPN expression in a blastocyst is regulated by estrogen through an estrogen receptor (ER). Our results of the delayed and activated implantation model showed that OPN expression is induced after estrogen injection. While estrogen treatment during embryo culture in vitro showed less effect on OPN expression, the tubal ligation model on day 3 of pregnancy confirmed that the regulation of estrogen on OPN expression in blastocyst might, through some specific cytokines, have existed in a uterine micro-environment. Collectively, our study presents that estrogen regulates OPN expression and it may play an important role during embryo implantation by activating blastocyst competence and facilitating the endometrium acceptable for active blastocyst.

[Luteal letrozole administration decreases serum estrogen level but not the risk of ovarian hyperstimulation syndrome].

OBJECTIVE: To explore the efficiency of using aromatase inhibitors during luteal phase in in vitro fertilization IVF stimulated cycles for patients at high risk for ovarian hyperstimulation syndrome (OHSS). METHODS: A total of 139 infertile women undergoing assisted reproductive technique with high risk for OHSS were enrolled in this clinical trial. In the treatment group 43 patients received five consecutive doses of aromatase inhibitors (letrozole) and support therapy combined with embryo cryopreservation. In the control group 96 patients received support therapy alone. All the patients were evaluated clinically, echographically, hematologically and tested for their steroid hormone. RESULTS: There was significantly lower estrogen level in the treatment group 2, 5 and 8 days after oocyte retrieval compared with the control group (P<0.001), There was no significant difference in luteinizing hormone and progesterone levels 2, 5 and 8 days after oocyte retrieval in the treatment group and control group (P>0.05). There were 7 cases of severe OHSS in the treatment group and 18 cases of severe OHSS in the control group. The rate of severe OHSS was not significantly different in the treatment group and control group (P=0.12). No side effect was reported in either group. CONCLUSION: Treatment with letrzolein luteal phase decreases serum estrogen levels of patients after oocyte retrieval,but it couldn't reduce the risk of severe OHSS.

[Ovarian torsion after controlled ovarian hyperstimulation: 5 cases report and clinical analysis].

OBJECTIVE: To evaluate the characteristics and treatment of ovary torsion after controlled ovarian hyperstimulation. METHODS: Between Jan.2008 and Dec.2011, 5 cases with ovary torsion who underwent ovarian hyperstimulation were retrospectively studied. RESULTS: Five cases presented intermittent lower abdominal from 1 to 38 days after oocyte retrieval. Enlargement of ovary and decreased or absent venous and/or arterial flow were demonstrated by Doppler sonography. Two torsions at left side, two torsions at right side, and one on bilateral side were observed. Three cases give up embryo transplantation, 2 cases were pregnant after surgical treatment. One case with partial torsion was successfully treated with simple conservative treatment. Two cases with complete torsion were performed adnexectomy by laparotomy. One case with complete torsion with early pregnancy was managed by laparoscopic adnexectomy. One case with chemical pregnancy was managed by laparoscopic detorsion for left side and excision for right side. Postoperative pathology of ovary tissue all confirmed haemorrhage and necrosis. CONCLUSIONS: Ovary torsion might occur after controlled ovarian hyperstimulation. The early management on ovary torsion will be benefit for preserving ovarian function.

[Osteopontin and male reproduction].

Osteopontin (OPN) is an extracellular matrix protein with multifunctions, expressed in various tissues and body fluids, involved in various physiological and pathological processes. It is also detected in the reproductive tract of both males and females, and participates in the implantation, development and differentiation of embryos. Recent studies have indicated that OPN is closely related with male fertility and may affect sperm quality and fertilization. An insight into the functions of OPN may help to explain the mechanisms of male infertility and improve the success rate of assisted reproductive technology.

[Expression of estrogen receptor, progesterone receptor and integrin alphaVbeta3 in endometrium of patients with luteinized unruptured follicle.].

OBJECTIVE: To investigate endometrium receptivity in patients with luteinized unruptured follicle (LUF) by measuring the expression of estrogen receptor (ER), progesterone receptor (PR) and integrin alphaVbeta3 in the endometrium. METHODS: From May 2007 to Nov. 2007, 17 infertile women with LUF were selected as LUF group matched with 13 infertile cases with normal ovulation as control group. They all underwent frozen-thawed embryo transfer in Reproductive Medicine Center, Renmin Hospital of Wuhan University. Endometrial tissue in anterior and posterior wall of uterus of LUF group and control group were biopsied by a small curettage between 7 and 11 days after luteinizing hormone (LH) surge. The expression of ER, PR and integrin alphaVbeta3 in endometrium were detected by immunohistochemistry staining. The level of estrogen and progesterone were measured by chemiluminescence assay. Then, the relationship between alphaVbeta3 expression in endometrium and the level of estrogen/progesterone were analyzed in LUF patients. RESULTS: (1) There was no remarkable difference in the level of estrogen between LUF [(656 +/- 299) pmol/L] and control group [(727 +/- 275) pmol/L, P > 0.05]. However, the level of progesterone were (23 +/- 8) nmol/L in LUF group and (35 +/- 10) nmol/L in control group, which reached statistical difference (P < 0.01). (2) The expression of ER, PR in endometrium of LUF patients were 183.9 +/- 2.4 and 168 +/- 3, which were significantly higher than 109.4 +/- 6.3 and 106 +/- 4 in control group (P < 0.01). The expression of integrin alphaVbeta3 in endometrium of 115 +/- 11 in LUF group were significantly lower than 191 +/- 9 in control group (P < 0.01). (4) In LUF group, the expression of alphaVbeta3 in endometrium was correlated positively with the level of progesterone (r = 0.77, P < 0.01) and irrelevant with the level of estrogen (r = 0.01, P > 0.05). CONCLUSION: The higher expression of estrogen and progesterone and lower expression of integrin alphaVbeta3 might confer impaired receptivity of endometrium and interfere with embryo implantation.

Effect of Progestin-primed Ovarian Stimulation Protocol on Outcomes of Aged Infertile Women Who Failed to Get Pregnant in the First IVF/ ICSI Cycle: A Self-controlled Study.

This study aimed to explore the outcomes of progestin-primed ovarian stimulation protocol (PPOS) in aged infertile women who failed to get pregnant in the first IVF/ICSI-ET cycles with GnRH-a long protocol. A self-controlled study was conducted to retrospectively investigate the clinical outcomes of 104 aged infertile patients who didn't get pregnant in the first IVF/ICSI-ET treatment by stimulating with GnRH-a long protocol (non-PPOS group), and underwent PPOS protocol (PPOS group) in the second cycle between January 2016 and December 2016 in the Center for Reproductive Medicine, Renmin Hospital of Wuhan University. The primary outcomes included clinical pregnancy rate of frozen-thawed embryos transfer (FET) in PPOS group, and good-quality embryo rate in both groups. The secondary outcomes were fertilization rate, egg utilization rate and cycle cancellation rate. The results showed that there were no significant differences in basal follicle stimulating hormone (bFSH), antral follicle count (AFC), duration and total dosage of gonadotropin (Gn), number of oocytes retrieved, intracytoplasmic sperm injection (ICSI) rate, fertilization rate, and cycle cancellation rate between the two groups (P>0.05). However, the oocyte utilization rate and good-quality embryo rate in PPOS group were significantly higher than those in non-PPOS group (P<0.05). By the end of April 2017,62 FET cycles were conducted in PPOS group. The clinical pregnancy rate and embryo implantation rate were 22.58% and 12.70%, respectively. In conclusion, PPOS protocol may provide better clinical outcomes by improving the oocyte utilization rate and good-quality embryo rate for aged infertile patients who failed to get pregnant in the first IVF/ICSI-ET cycles.

Fyn Plays a Pivotal Role in Fetomaternal Tolerance Through Regulation of Th17 Cells.

PROBLEM: Spontaneous abortion is a poorly understood phenomenon, although fetomaternal intolerance is known to play an important role in its pathogenesis. The tyrosine-specific phosphotransferase Fyn has been reported as a significant regulator in immune response. However, its role in fetomaternal immune tolerance and contribution to spontaneous abortion remains unclear. METHODS OF STUDY: Fyn expression was evaluated at the fetomaternal interface of normal pregnant and abortion-prone mice, as well as in decidual tissue obtained from normal human pregnancies and idiopathic miscarriages. A Fyn inhibitor was administrated into the LPS-induced abortion mice to investigate the variation of embryo resorption and local immunity. RESULTS: Fyn expression fluctuated with the progress of normal pregnancy and was elevated in abortion-prone mice and patients with recurrent spontaneous abortion. The Fyn inhibitor reversed LPS-induced embryo absorption and aberrant inflammatory status including redundant expression of IRF4 and increased proportion of Th17 cells. CONCLUSION: Fyn is confirmed as a negative regulator in fetomaternal immune tolerance, through promoting Th17 cell expansion and proinflammatory factors expression.

Patients with cervical Ureaplasma Urealyticum and Chlamydia Trachomatis infection undergoing IVF/ICSI-ET: The need for new paradigm.

Genital tract infections with ureaplasma urealyticum (UU) and chlamydia trachomatis (CT) are the most frequent sexually-transmitted disease worldwide. UU and CT infections are considered to be the leading cause for infertility and adverse pregnancy outcomes. However, little is known about the specific effect of cervical UU and CT infections on the etiology of female infertility, as well as the pregnancy outcomes of the patients undergoing in vitro fertilization/intracytoplasmic sperm injection-embryo transfer (IVF/ICSI-ET). In order to find the association between cervical UU and/or CT infection and pregnancy outcomes, we conducted a retrospective case-control study on the patients undergoing IVF/ICSI-ET with cervical UU and/or CT infection. A total of 2208 patients who received IVF/ICSI-ET were enrolled in this study. Data on the general conditions, pregnancy history and clinical pregnant outcomes were analyzed in terms of the cervical UU and CT detection. Our results revealed that cervical UU and CT infections were the risk factors for ectopic pregnancy and tubal factor-induced infertility. Moreover, the pregnancy rate, abortion rate, ectopic pregnancy rate and premature birth rate in patients with UU and/or CT infections showed no significant difference when compared with the control group. We recommend that cervical UU and CT detection should be an optional item for infertility patients and clinical UU detection should differentiate the subtypes of cervical UU. Positive cervical UU and CT infections should not be taken as strict contraindications for IVF/ICSI-ET.

Osteopontin facilitates invasion in human trophoblastic cells via promoting matrix metalloproteinase-9 in vitro.

Successful implantation of embryo and placentation depend on proper trophoblast proliferation and differentiated into specialized invasive trophoblast. However, little is known about the regulatory factors and mechanisms in trophoblast proliferation and differentiation. Osteopontin (OPN) is a member of the small integrin-binding ligand N-linked glycoprotein family and participates in cell adhesion and invasion. It has been identified that OPN is highly expressed in invasive trophoblasts in human placenta. In this study, we demonstrated that OPN is constitutively expressed in highly invasive phenotype of human choriocarcinoma cell lines of JAR and JEG-3 cells, and OPN could promote trophoblast proliferation and invasion, partly through promoting MMP-9 secretion. Inhibition of OPN will compromise the abilities of proliferation and invasion in JAR and JEG-3 cell lines. Our data showed that the expression of OPN in trophoblast may participate in placentation, OPN expression defects may be involved in gestational trophoblastic diseases.

[Effects of intrauterine contraceptive device on expression of vascular endothelial growth factor, kinase insert domain-containing receptor and microvessel density in endometrium].

OBJECTIVE: To investigate the expression of vascular endothelial growth factor (VEGF) and its receptor, kinase insert domain-containing receptor(KDR) and microvessel density (MVD) in endometrium from women wearing fixed copper-intrauterine contraceptive device (IUD, FCu-IUD) or fixed indomethacin-releasing copper-IUD (FICu-IUD). METHODS: Twenty healthy women were divided into two study groups: 10 cases wearing the FCu-IUD (FCu-IUD group), 10 cases wearing the FICu-IUD (FICu-IUD group). Immunohistochemical technique was used to determine the expression of VEGF and KDR in endometrium, and the microvessel density (MVD) was counted. The expression of VEGF mRNA was determined by in situ-hybridization. RESULTS: Before insertion of FCu-IUD, the expression of VEGF and KDR proteins was 0.357 +/- 0.032 and 0.215 +/- 0.029 respectively. After insertion of FCu-IUD, the expression of VEGF and KDR proteins was 0.568 +/- 0.027 and 0.244 +/- 0.022 respectively, significantly higher than before insertion (P < 0.05). The expression of VEGF mRNA was 0.359 +/- 0.022 before insertion of FCu-IUD, after insertion of FCu-IUD, the expression of VEGF mRNA was 0.425 +/- 0.019 (P < 0.05). There were no significant changes in the level of VEGF protein and mRNA, as well as KDR in endometrium before and after insertion of FICu-IUD. Compared with before insertion of FCu-IUD 15.4 +/- 2.8, a significant increase in MVD was observed after insertion of FCu-IUD 19.8 +/- 4.8, and the expression of VEGF protein was positively correlated with MVD (r = 0.847, P < 0.01). MVD counts were not different significantly before and after insertion of FICu-IUD. CONCLUSIONS: FCu-IUD can enhance the expression of VEGF and KDR in the endometrium. FICu-IUD can inhibit the activity of VEGF and KDR by releasing indomethacin. VEGF and KDR may be related to the structural and functional changes of microvessels in endometrium after insertion of FCu-IUD or FICu-IUD.

Osteopontin is expressed in the mouse uterus during early pregnancy and promotes mouse blastocyst attachment and invasion in vitro.

Embryo implantation into the maternal uterus is a decisive step for successful mammalian pregnancy. Osteopontin (OPN) is a member of the small integrin-binding ligand N-linked glycoprotein family and participates in cell adhesion and invasion. In this study, we showed that Opn mRNA levels are up-regulated in the mouse uterus on day 4 and at the implantation sites on days 5 and 8 of pregnancy. Immunohistochemistry localized the OPN protein to the glandular epithelium on day 4 and to the decidual zone on day 8 of pregnancy. OPN mRNA and proteins are induced by in vivo and in vitro decidualization. OPN expression in the endometrial stromal cells is regulated by progesterone, a key regulator during decidualization. As a secreted protein, the protein level of OPN in the uterine cavity is enriched on day 4, and in vitro embryo culturing has indicated that OPN can facilitate blastocyst hatching and adhesion. Knockdown of OPN attenuates the adhesion and invasion of blastocysts in mouse endometrial stromal cells by suppressing the expression and enzymatic activity of matrix metalloproteinase-9 in the trophoblast. Our data indicated that OPN expression in the mouse uterus during early pregnancy is essential for blastocyst hatching and adhesion and that the knockdown of OPN in mouse endometrial stroma cells could lead to a restrained in vitro trophoblast invasion.