BMP7 coordinates endometrial epithelial cell receptivity for blastocyst implantation through the endoglin pathway in cell lines and a mouse model.

Bone morphogenetic protein (BMP) expression has been observed in the uterus in previous studies. However, the influence of BMP7 on blastocyst implantation remains unclear. Blastocysts first act on luminal endometrial epithelial cells during implantation. The purpose of the present study was to explore the influence of BMP7 on endometrial epithelial cells. A pregnancy animal model, and mouse and human endometrial epithelial cells were used in the present study. Transient knockdown, immunofluorescence assay, in vitro embryo implantation, BMP7 silencing, reverse transcription-quantitative polymerase chain reaction, western blotting, immunoprecipitation and Rac1 function assay were also performed. It was revealed that BMP7 concentration was increased in endometrial epithelial cells during the final pre-receptive and receptive stages of receptivity in the mouse endometrium. Additionally, BM7 acted on the transforming growth factor-ß receptor, endoglin. Endoglin expression was detected in both stromal and endothelial cells apart from trophoblast expression. Following knockdown of BMP7, Rac-GTP was decreased in endometrial epithelial cells and the uterus. Knockdown of endoglin by small interfering RNA decreased the number of blastocysts and implantation regions. Additionally, BMP7 silencing and endoglin suppression of Ishikawa cells led to impaired JAr spheroid attachment. These findings suggest that BMP7 is associated with receptivity of the endometrium, indicating that BMP7 regulates receptivity of endometrial epithelial cells for implantation of blastocysts via the endoglin pathway.

The Effect of Atosiban on Patients With Difficult Embryo Transfers Undergoing In Vitro Fertilization-Embryo Transfer.

The purpose of this study was to evaluate the effect of atosiban on the outcomes of infertile women undergoing in vitro fertilization (IVF) with difficult embryo transfers (ETs). This randomized double-blind study enrolled 204 infertile women with difficult ETs during IVF treatment between June 2014 and June 2018. According to a computer-generated randomization list, participants were randomized into placebo (n = 102) and atosiban (n = 102) groups. In atosiban group, atosiban with a total dose of 37.5 mg was administered. All of the patients underwent IVF-ET using cryopreserved embryos. The clinical pregnancy rate per cycle and implantation rate per transfer (45.1% and 26.5%) in atosiban group were significantly higher than those of placebo group (15.6% and 9.7%, respectively; P < .05). This study showed that administration of atosiban during ET was extraordinarily effective for patients with difficult transfers.

5-Aminolevulinic acid photodynamic therapy in human cervical cancer via the activation of microRNA-143 and suppression of the Bcl-2/Bax signaling pathway.

5-Aminolevulinic acid photodynamic therapy (ALA­PDT) is a method using a photosensitizer and light radiation for disease treatment, and is currently used for the treatment of skin cancers, precancerous lesions and viral warts. The present study aimed to investigate the effect of ALA­PDT on human cervical cancer through the regulation of microRNA­143 (miR­143) and the Bcl-2/Bax signaling pathway. The results demonstrated that ALA­PDT reduced proliferation, increased cytotoxicity and induced apoptosis in HeLa human cervical carcinoma cells. Reverse transcription­quantitative polymerase chain reaction analysis demonstrated that the expression levels of miR­143 were increased following ALA­PDT treatment. Western blotting indicated that the expression levels of Bcl­2 and Bax were significantly reduced and increased, respectively, by ALA­PDT treatment. In addition, upregulation of miR­143 expression reduced Bcl­2 expression and increased Bax expression in HeLa cells. However, downregulation of miR­143 expression inhibited the effect of ALA­PDT on Bcl-2/Bax protein expression. In conclusion, the current study demonstrated that ALA­PDT affected human cervical cancer via the activation of miR-143 and the suppression of the Bcl-2/Bax signaling pathway.

Effect of Indoleamine 2,3-Dioxygenase Expressed in HTR-8/SVneo Cells on Decidual NK Cell Cytotoxicity.

PROBLEM: To study the effect of indoleamine 2,3-dioxygenase (IDO) expressed in HTR-8/SVneo cells on NKG2D and NKp46 expression and cytotoxicity of decidual NK (dNK) and peripheral NK (pNK) cells. METHOD OF STUDY: CD56(+) dNK and pNK cells purified were cultured with HTR-8/SVneo cell conditioned medium (CM), 1-MT+HTR-8/SVneo cell CM, and complete RPMI 1640 medium (negative control) in vitro. The mRNA and protein expression of NKG2D and NKp46 in NK cells were then assessed by qRT-PCR and flow cytometry, respectively. Their cytotoxicity was evaluated with LDH assays, and TNF-α secretion was analyzed by ELISA. RESULTS: For dNK cells, the mRNA and protein expression of NKp46 as well as NKG2D did not differ significantly among the three groups (P > 0.05), whereas for pNK cells, the expression level was significantly decreased in HTR-8/SVneo cell CM group than the other two groups (P < 0.01). Peripheral NK cells cultured with HTR-8/SVneo cell CM showed reduced cytotoxicity and TNF-α secretion than the other two groups (P < 0.01), although there were no significant differences among three groups for dNK cells (P > 0.05). CONCLUSION: IDO expressed by HTR-8/SVneo cells can down-regulate NKp46 and NKG2D expression and reduce cytotoxicity in pNK cells, and may contribute to keep dNK cytotoxicity at a low level, suggesting an important role for IDO in the maintenance of normal pregnancy.

Morphological and Ultrastructural Changes in Human Endometrium Following Low-Dose Levonorgestrel Contraceptive Intrauterine Systems (LNG-IUS-12) 13.5 mg.

OBJECTIVE: To study the effect of low-dose levonorgestrel contraceptive intrauterine systems (LNG-IUS-12) on endometrial morphology and ultrastructure. METHODS: Twenty seven women who desired contraception were inserted LNG-IUS-12. Endometrial biopsies were collected during the late proliferative phase of the cycle before (control) and after the use of the LNG-IUS for 36 months. The changes of morphology and ultrastructure of endometrium tissue were observed by light microscope and transmission electron microscope. RESULTS: After 36 months of use, the endometrial gland reduced in number and some were atrophic. A decidual change of stroma was present. Electronic microscopy demonstrated the presence of low columnar, karyopyknosis, and chromatin margination in the glandular epithelial cell. Some vacuolar degeneration in partial mitochondria and rough endoplasmic reticulum presented light extension. Changes could be concluded as cell inhibition of proliferation. CONCLUSIONS: The low-dose levonorgestrel-releasing intrauterine systems has a suppressive effect on endometrium that is similar to LNG-IUS.

[Efficacy and safety of mifepristone combined with misoprostol for termination of pregnancy between 8 and 16 weeks of gestation].

OBJECTIVE: To assess the efficacy and safety of mifepristone combined with oral or vaginal misoprostol for termination of pregnancy between 8 and 16 weeks of gestation. METHODS: This was a randomized, multi-center, open clinical trial. A total of 625 women at 8-16 weeks of gestation were randomized to receive 200 mg oral mifepristone followed by either oral misoprostol 400 µg every 3 hours or vaginal misoprostol 400 µg every 6 hours for a maximum of 4 doses 36-48 hours later. There were 417 women in oral group with 198 at 8-9 weeks and 219 at 10-16 weeks, while 208 women in vaginal group with 99 at 8-9 weeks and 109 at 10-16 weeks. The outcome measures were the success abortion rate, induction to abortion interval, the amount of bleeding, reoccurrence of menstruation and adverse events. RESULTS: Abortion rate was significantly higher in vaginal group [98.1% (202/206)] than that in oral group [94.0% (390/415), P = 0.023]; concerning termination of pregnancy at 8-9 weeks and 10-16 weeks respectively, there were no significant differences between oral and vaginal groups (P = 0.156, P = 0.073). The induction to abortion interval was no significant difference in oral and vaginal group in different gestational weeks (P = 0.238, P = 0.273). The average induction to abortion interval was (4.1 ± 6.6) hours and (6.0 ± 4.5) hours respectively in terminating 8-9 weeks and 10-16 weeks of gestation. Concerning the amount of bleeding within 2 hours of placenta expulsion, there was significant difference between oral group [(63 ± 46) ml] and vaginal group [(55 ± 45) ml] in terminating 8-9 weeks of gestation (P = 0.047), while there was no significant difference between groups in terminating 10-16 weeks of gestation [oral group (76 ± 52) ml versus vaginal group (76 ± 61) ml, P = 0.507]. The reoccurrence of menstruation was about 37 days in both oral and vaginal groups. Two cases of incomplete abortion were serious adverse events (SAE) relating to treatment. The common adverse events (AE) of nausea and vomiting were significantly higher in oral group [57.2% (239/417), 36.3% (151/417)] than those in vaginal group [45.4% (94/208), 26.1% (54/208); P = 0.005, 0.011]. CONCLUSION: Oral or vaginal misoprostol combined with mifepristone, is effective and safe for termination of pregnancy between 8 and 16 weeks of gestation.

Delayed presentation of uterine perforation with ovary migration after dilatation and curettage.

We present a rare but serious uterine perforation. A 31-year-old woman was referred to our department for hyperechogenic mass in uterus on ultrasonography after Dilation and curettage (D&C) for the adherent placenta and retained products of conception. Transvaginal ultrasound examination showed that a mass with several follicles measuring 35×29 mm was seen emanating from the right posterior wall of the uterine cavity, and there was absence of the myometrial tissue. A hysteroscopy and laparoscopy showed a uterine perforation with ovary incarceration. The ovary was rehabilitated, and the uterine perforation site was incised. D&C can not be performed when delayed presentation of uterine perforation with migration of an extrauterine organ is suspected, particularly, some of them are asymptomatic after a difficult intrauterine operation.

[Relative factors of expectant management of ectopic pregnancy].

OBJECTIVE: To analyze select patients of ectopic pregnancy for expectant management. METHODS: Retrospective analyses were conducted for the relative factors of 180 cases of ectopic pregnancy with expectant management at Department of Family Planning of our hospital from August 2004 to January 2012. Their general clinical data, clinical manifestations and laboratory tests were collected. The cases requiring neither surgery nor drug therapy belonged to the cure group while the rest fell into the failure group. RESULTS: A total of 140 patients were successfully managed with a cure rate of 75.27%. There was no significant difference between two groups in size of mass on ultrasonography (P > 0.05). The cases with gestational sac of mass on ultrasonography had statistical difference between two groups (P < 0.05). Statistical difference existed between two groups (P < 0.01) in initial blood beta-human chorionic gonadotropin (ß-hCG), initial bloodß-hCG and D3-5/D0 (ratio of bloodß-hCG at day 3 - 5 and initial) were the variables for predicting the likelihood of successful expectant management under the ROC curve. The Youden Index was the largest at an initial bloodß-hCG of 634 IU/L or D3-5/D0 of 0.711. CONCLUSION: At blood ß-hCG ≤ 634 IU/L or D3-5/D0 < 0.711, expectant management may be offered. And mass with gestational sac on ultrasonography is a relative factor of expectant management.

Indoleamine 2,3-dioxygenase levels at the normal and recurrent spontaneous abortion fetal-maternal interface.

OBJECTIVES: To localize indoleamine 2,3-dioxygenase (IDO) mRNA and protein and to undertake a functional study at the first trimester fetal-maternal interface in order to determine whether the distribution and function of IDO are related to recurrent spontaneous abortion (RSA). METHODS: Women undergoing legal pregnancy termination and women with RSA participated in this prospective study. Immunohistochemistry and real-time reverse transcription-polymerase chain reaction were used to analyse levels of IDO protein and mRNA in placenta, decidua and HTR-8/SVneo cells. Culture medium collected from trophoblast villous explant or HTR-8/SVneo cell cultures was used to measure IDO activity in response to interferon (IFN)-γ treatment. RESULTS: A total of 40 healthy women and 26 women with RSA provided samples of placenta and decidua. For normal pregnancies, IDO protein and mRNA was identified in placental trophoblasts, invasive extravillous trophoblasts and decidual glandular epithelium. IFN-γ significantly increased IDO activity in trophoblast villous explants and HTR-8/SVneo cells. Levels of IDO protein and mRNA in the placenta and decidua from normal pregnancies were significantly higher than in those from RSA. CONCLUSIONS: Decreased levels of IDO protein and mRNA in the placenta and decidua from RSA suggest an important role for IDO in the maintenance of normal pregnancy.

Expansion of decidual CD45RO⁺ T cells with high expression of CEACAM1 in the early stage of pregnancy.

The aim of the present study was to investigate the mechanism involved in the expansion of CD45RO+ T cells in the decidual microenvironment, and in the expression of the inhibitory carcinoembryonic antigen-related cell adhesion molecule 1 (CEACAM1) on the surface of decidual CD45RO+ T cells. Twenty-one healthy nonpregnant females and seventeen healthy pregnant females in the first trimester were included in the study. Peripheral blood samples from nonpregnant and pregnant females, and decidual tissues from pregnant females following elective abortion, were obtained and analyzed by flow cytometry. The percentages of CD45RO+ T cells and CEACAM1-expressing CD45RO+ T cells were significantly higher in first trimester human decidua than in the peripheral blood. Conditioned medium from the coculture of monocytes and the human trophoblast HTR8/SVneo cell line (MHM) was added to the model for the generation of CD45RO+ T cells in vitro. MHM caused an increase in the percentage of CD45RO+ T cells in a monocyte chemoattractant protein-1 (MCP-1)­dependent manner and an increase in the percentage of CEACAM1-expressing CD4+CD45RO+ T cells in the model. In conclusion, our results implied that trophoblast cells and monocytes may be involved in the increase of decidual CD45RO+ T cells and the high expression of CEACAM1 on their surfaces.

A novel medical device for removal of intrauterine devices under direct vision.

BACKGROUND: Evaluation of the efficacy of a novel medical device for removal of retained intrauterine devices (IUDs) under direct vision without uterine distention is reported. CASES: The five cases presented used the novel medical device to remove the IUDs. One woman had an incarcerated IUD, two women had fractured IUDs and two women had IUDs with lost threads. All IUDs were removed completely with use of direct vision without uterine distention. CONCLUSIONS: The device is effective for the removal of incarceration IUDs under direct vision without uterine distention.

[Comparison of ultrasound plus radiography versus computed tomography in the diagnosis of ectopic intrauterine devices].

OBJECTIVE: To compare the effectiveness of ultrasound plus radiography versus computed tomography (CT) in the diagnosis of ectopic intrauterine device (IUD) and analyze the ratio of cost and effectiveness (C/E) so as to provide scientific rationales for the selection of appropriate diagnostic methods. METHODS: A total of 70 cases were recruited from two major Chinese hospitals in 2009. They were examined with ultrasound plus radiography and CT respectively. The gold diagnostic criterion was postoperative diagnosis to compare the sensitivity, specificity, diagnostic index (DI), accuracy and C/E for different diagnostic methods. The SPSS 13.0 statistical analysis software was employed for data analysis. RESULTS: The data of 65 subjects were collected and analyzed. For ultrasound plus radiography, the sensitivity, specificity, DI, accuracy and C/E were 82.1%, 88.9%, 171.0%, 83.1% and 137.3 respectively. As for CT, the above indices were 96.4%, 55.6%, 152.0%, 90.8% and 170.7 respectively. CONCLUSION: Considering the higher levels of specificity and DI and a lower C/E, ultrasound plus radiography is superior to CT so that the former modality shall become a first-choice in the diagnosis of ectopic IUD, especially at the grass-root family planning service stations.

Angiographic uterine artery chemoembolization followed by vacuum aspiration: an efficient and safe treatment for managing complicated cesarean scar pregnancy.

PURPOSE: To evaluate uterine artery chemoembolization (UAEC) followed by vacuum aspiration as a conservative treatment for complicated cesarean scar pregnancy. METHODS: A retrospective review was performed of women presenting with cesarean scar pregnancy between January 2002 and December 2008. The medical record was evaluated to determine the method of treatment. RESULTS: During the time period studied, 13 women were identified who underwent UAEC followed by vacuum aspiration. 12 women successfully had bilateral UAEC followed by vacuum aspiration alone, one woman had unilateral UAEC followed by vacuum aspiration but subsequently required laparotomy. All 13 women were successfully cured and retained uterus, there was no case with severe complicating disease. With the follow-up period, two women who were planning future pregnancy conceived, and spontaneous abortion occurred in one of them during the first trimester, another had an elective cesarean delivery at term. CONCLUSIONS: UAEC combined with vacuum aspiration is technically feasible and may help avoid laparotomy in women with cesarean scar pregnancy.

Timing and indication for curettage after medical abortion in early pregnant women with prior uterine incision.

BACKGROUND: Termination of pregnancy is an important and necessary back-up method for family planning services in many countries. The combination of mifepristone and misoprostol is a widely used alternative to surgical evacuation of the uterus in early pregnancy; however, there are few reports about medical abortion in women with a prior uterine incision and few studies have described curettage occurring as part of the procedure and an indication for the intervention. Curettage in a prior uterine incision can increase operative complications. The purpose of this study was to investigate whether vaginal bleeding intervals, routine ultrasound scan and serum beta-hCG test after medical abortion could accurately identify women with uterine scars who would require curettage. METHODS: Six hundred sixty-eight women with a uterine scar and at up to 49 days of gestation underwent a medical abortion with mifepristone and misoprostol. Each woman took 50 mg and 25 mg of mifepristone orally in the morning and in the evening, respectively, for 2 days and 600 mcg of misoprostol orally on the third day. RESULTS: Of the 668 women, 6 (0.9%) were lost to follow-up. The overall complete abortion rate was 91.7%; 55 women underwent curettage, including 2 women with heavy bleeding, 3 women with ongoing pregnancy and 34 women with incomplete abortion. The incomplete abortion rate was significantly greater in women with persistent bleeding lasting 21 days than in women with persistent bleeding lasting 14 days (p<.001), and the overall sensitivity and specificity of vaginal bleeding interval (21 days) were 97.1% and 75%, respectively. The incomplete abortion rate was also greater in women whose serum beta-hCG was >or=500 IU/L than in women whose serum beta-hCG was <500 IU/L (p<.001), and the overall sensitivity and specificity of serum beta-hCG (>or=500 IU/L) were 97.1% and 62.5%, respectively. Moreover, the incomplete abortion rate was greater in women with an endometrial thickness >or=15 mm than in women with an endometrial thickness <15 mm (p<.001), and the overall sensitivity and specificity of endometrial thickness (>or=15 mm) were 94.1% and 75%, respectively. No complication occurred. CONCLUSIONS: The combination of mifepristone and misoprostol was found to be a safe and effective method to terminate early pregnancy in women with a previous cesarean delivery. If a woman with a prior uterine incision experienced vaginal bleeding intervals >or=21 days and/or had a bilayer endometrial thickness >or=15 mm and/or serum beta-hCG >or=500 IU/L after a medical abortion, then she should undergo curettage.

Biological characteristics of human placental mesenchymal stem cells and their proliferative response to various cytokines.

The placenta is an attractive new source of mesenchymal stem cells (MSCs), but the biological characteristics of placenta-derived MSCs (P-MSCs) have not yet been characterized. We successfully isolated, cultured and expanded P-MSCs using routine methods. Under appropriate induction conditions, these cells can differentiate into bone, cartilage, fat and hepatocyte-like cells. In addition, the proliferative response of P-MSCs to different cytokines was monitored using the MTT assay. The results show that low concentrations of proinflammatory cytokines, e.g. RANTES, interleukin (IL)-1, IL-6 and IL-8 can stimulate the proliferation of P-MSCs in a dose-dependent manner, peaking at concentrations of 40 ng/ml of RANTES, 10 ng/ml of IL-1 and IL-6, and 150 ng/ml of IL-8 (p < 0.01). The level of proliferation decreased when the concentration of these four cytokines increased beyond these values. On the other hand, anti-inflammatory cytokines hepatocyte growth factor and IL-4 had an inhibitory effect on P-MSCs. In conclusion, the placenta contains MSCs that are consistent with the characteristics of bone marrow MSCs. Low concentrations of proinflammatory chemokines stimulated the proliferation of P-MSCs while anti-inflammatory cytokines inhibited the growth of P-MSCs in a dose-dependent manner.

[Effects of long-term installation of intrauterine devices on intrauterine microenvironment].

OBJECTIVES: To study the effect of long-term installation of intrauterine devices (IUD) on the intrauterine microenvironment. METHODS: Eighty-nine healthy 26 - 50-year-old women undergoing physical examination or having their IUD removed were recruited. Among them 62 had used IUDs, including 32 inert IUD (I-IUD) and 30 copper releasing IUD (T-IUD), for 5 - 20 years, and 27 women without installation of IUD were used as controls. In the 3rd to 13th day of menstrual cycle, 3 ml of irrigation of intrauterine cavity were collected to examine the concentrations of tumor necrosis factor-alpha (TNF-alpha), interleukin 2 (IL-2) and secretary IgA (SIgA) by radioimmunoassay (RIA). Hysteroscopy was used to obtain 3 pieces of endometrium at the place where the IUD was attached for each subject. Three pieces of endometrium were curretted from each control during the proliferative stage. Thirty specimens of endometrium, including 10 cases with I-IUD, 10 cases with T-IUD, and 10 control cases were used to examine the distribution of T cell subset by immunohistochemistry. Fifteen specimens of endometrium, including 6 cases with I-IUD, 6 cases with T-IUD, and 3 control cases, were used to examine the endometrial ultrastructure by scanning electron microscopy (SEM) and transmission electron microscopy (TEM). Twenty-five specimens of endometrium, including 10 cases with I-IUD, 10 cases with T-IUD, and 5 control cases, were used to detect the expression and mutation of P16, P53 and K-ras by PCR-SSCP technique. RESULTS: There was no significant difference in TNF-alpha level in the irrigation among the three groups (P > 0.05). IL-2 and SIgA levels (0.39 +/- 0.18) microg/L and (2000 +/- 1224) microg/L respectively) in the T-IUD group were significantly lower than those in the control group (0.96 +/- 0.15) microg/L and (3377 +/- 1906) microg/L respectively, both (P < 0.05). There was a significant difference in the population of T lymphocytes (CD(8)(+) and CD(4)(+) lymphocytes) between the T-IUD group and control group (P < 0.05). SEM and TEM showed no necrosis and atypia in the endometrial cells of I-IUD group and T-IUD group. There was no positive expression of gene p16, p53 and K-ras in the endometrium of the three groups. CONCLUSION: It is safe and effective using I-IUD or T-IUD for a long time, however, T-IUD has some effects on intrauterine local immune function.