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Background: The previous model-based cost-effectiveness analyses regarding elective oocyte cryopreservation remains debatable, while the usage rate may influence the price per live birth. The aim of this study is to disclose the usage and cost-effectiveness of the planned cryopreserved oocytes after oocyte thawing in real-world situations in which in-vitro fertilization (IVF) is exclusively provided to married heterosexual couples. Methods: This was a retrospective single-center observational study. Women who electively cryopreserved oocytes and presented to thaw the oocytes were categorized as thawed group. The oocytes were fertilized at our center and the sperm samples for each individual was retrieved from their respective husbands. Clinical outcomes were traced and the live birth rate per thawed case was calculated. The costs from oocyte freezing cycles to oocyte thawing, and embryo transfer cycles were accordingly estimated. Cumulative costs per live birth was defined by the cumulative costs divided by the total live births per thawed case.Results: We recruited 645 women with 840 oocyte retrieval cycles for elective oocyte freezing from November 2002 to December 2020. The overall usage rate was 8.4% (54/645). After the storage duration exceeded 10 years, the probabilities of thawing oocytes were 10.6%, 26.6%, and 12.7% from women who cryopreserved their oocytes at the age of ≤35 years, 36–39 years, and ≥40 years, respectively. Among women who thawed their oocytes, 31.5% (17/54) of women achieved live births. For the age groups of ≤35 years, 36–39 years, and ≥40 years, the total live birth rate per thawed case was 63.6%, 42.3%, and 17.6%, respectively, and the cumulative costs for one live birth was $11,704, $17,189, and $35,642, respectively.Conclusions: The overall usage rate was 8.4% in our cohort. The chance of having live births was 31.5% if the individual thawed their oocytes. The findings added to the limited evidence of the usage rate in real-world situations in which IVF is exclusively available to married heterosexual couples, which could aid future analysis and decision-making for women willing to preserve fertility to avoid age-related fertility loss.Trial Registration Number: None.

BackgroundEstrogen (E2) is a key regulator of trophoblast invasion and remodeling of uterine decidual spiral arterioles during placentation. However, there is little research on the effects of E2 in early pregnancy on placentation and subsequent preeclampsia. This study aims to evaluate the relationship between the rate of hypertensive disorders of pregnancy (HDP) and serum estrogen levels in early pregnancy following frozen embryo transfer (FET).MethodsThis retrospective study included all patients who conceived after FET with singleton delivery in the National Taiwan University Hospital from January 1, 2006, to December 31, 2020. A total of 729 patients were included in the analysis. The primary outcome was the rate of HDP.ResultsSerum estrogen levels at the fourth and fifth gestational weeks were significantly different between women with and without HDP after FET (393.4 ± 235.7 vs 479.0 ± 261.2 pg/ml, p=0.007 and 492.0 ± 274.5 vs 563.9 ± 263.0 pg/ml, p=0.015, respectively). Multivariable logistic regression with generalized estimating equations showed that each 100-pg/mL increase in estrogen levels increased the risk of HDP by 12% (p = 0.006) after adjustment for confounders. In a subanalysis stratified using FET protocol type (natural cycle, n = 225; hormone replacement cycle, n = 504), an increased rate of HDP (P-trend = 0.021 and 0.014, respectively) was observed with increasing estrogen level quartile at the fourth and fifth gestational weeks in the hormone replacement cycles, but not in the natural cycles.ConclusionsHigher estrogen levels in early pregnancy were associated with a higher rate of HDP. For better obstetric outcomes, we may alter the estradiol supplementation in FET hormone replacement cycles or choose FET natural cycles. Our results indicate a link between estrogen levels in early pregnancy and the rate of HDP. Further studies are needed to investigate the underlying mechanism.Trial registrationRetrospectively registered

This study aims to compare the efficacy, tolerability and patient satisfaction between aqueous subcutaneous progesterone (Prolutex, 25 mg/vial; IBSA) and vaginal progesterone (Crinone, 90 mg/tube; Merck) as luteal support for fresh embryo transfers in in-vitro fertilization (IVF).In this prospective randomized study, 65 patients who underwent IVF were recruited and randomly assigned to either the Prolutex (25 mg daily, n = 33) or Crinone (90 mg daily, n = 32) group. The luteal support regimens were given daily, starting from two days after oocyte pickup. If the serum pregnancy test was positive, luteal support was continued until 7 weeks of gestation. Primary outcomes were clinical pregnancy rate and serum progesterone level at the mid-luteal phase and at 4 weeks of gestation. Secondary outcomes were drug tolerability and patient satisfaction assessed by questionnaire.There were no significant differences in clinical pregnancy rates (Prolutex 25.0% versus Crinone 33.3%, p = 0.699), serum progesterone levels and patient satisfaction between Prolutex and Crinone group. Although the patients that had received Prolutex complained of more local pain at the injection sites, they also had less annoying vaginal discharges and vulvar discomforts.Prolutex is of comparable efficacy and patient satisfaction to Crinone, and its availability means patients have more options in regards to the routes of progesterone administration as luteal phase support during IVF.

Objective: To study the impact of stimulation duration on intracytoplasmic sperm injection (ICSI) - embryo transfer (ET) outcome in poor and normal responders during controlled ovarian stimulation using gonadotropin-releasing hormone (GnRH) antagonist protocol. Materials and methods: This is a retrospective cohort study. There were 1481 women undergoing ICSI-ET cycles. Women with ovum pick-up number ≤3 were defined as poor responders (n = 235), and those with a number ≥4 were normal responders (n = 1246). Results: The mean stimulation duration was shorter in poor responders with pregnancy group as compared with normal responders with pregnancy group (7.8 ± 2.2 vs. 9.2 ± 1.6 days, p

Previous studies indicated that progesterone can be withdrawn at the time of the first positive β-hCG test without compromising the clinical pregnancy outcome in normal ovarian responder. However, the effect of early stop of progesterone supplementation for patients with poor ovarian response (POR) has not been investigated. This study retrospectively collected data from patients with POR in 156 IVF/ICSI fresh embryo transfer (ET) cycles in single tertiary center from January 2010 to June 2016. All the patients met ESHRE consensus, the Bologna criteria, of POR and had hCG injection for luteal phase support (LPS) on day 2, 5 and 8 after ovum pick-up. The pregnant patients were divided into two groups: early stop group represented those who stopped LPS from day of positive pregnancy test; control group represented those who kept progesterone supplementation till gestational age of 9 weeks. There were no significant differences in age, BMI, parity, hormone data, number of follicles>10(mm), endometrial thickness and number of embryos transferred between the two groups. After adjustment for possible confounders with multivariate logistic regression analysis, the clinical pregnancy rates (55.0% vs. 57.1%, P = 0.35), ongoing pregnancy rates (47.0% vs. 46.4%, P = 0.66), miscarriage rates (34.0% vs. 26.7%, P = 0.66) and live-birth rates (44.0% vs. 46.4%, P = 0.41) were not statistically different between early stop group and the control group. Our study indicates that early stop of progesterone supplementation on the day of positive pregnancy test for patients of POR using hCG as LPS in fresh ET cycles does not affect pregnancy outcome.

Background/Purpose: The role of LH during controlled ovarian stimulation (COS) in the general population remains contentious. There is no consensus on the indications for LH supplementation during COS. The purpose of this study is to determine whether menotropin supplement is associated with decreases in early pregnancy loss rates in patients exhibiting low endogenous LH during COS. Method: This is a single-center, retrospective cohort from a university-affiliated hospital. Patients were enrolled from the in-vitro fertilization center from January, 2011 to December, 2014. Patients who experienced a LH level ≦ 0.8 mIU/mL during stimulation were identified, and patients that received menotropin supplementation were compared to those without menotropin supplementation. Outcome variables, including the number of oocytes retrieved, embryos obtained, implantation rates, pregnancy rates and early pregnancy loss rates, were compared. Results: Patients that experienced low LH during GnRH antagonist protocol and were supplemented with menotropin were associated with lower early pregnancy loss when compared with patients without menotropin supplementation (26.7% vs. 11.5%, p = 0.045). More specifically, in patients who exhibited early-onset low LH, before the use of GnRH antagonists, menotropin supplementation was associated with significantly lower early pregnancy loss compared with non-supplemented patients (3.3% vs. 29.0%, OR: 0.08, p = 0.012). Beneficial effects persisted after adjusting for confounders (aOR: 0.103, 95% CI: 0.011–0.933). Conclusion: Menotropin supplementation is associated with decreased early pregnancy loss in patient who exhibited low LH during GnRH antagonist cycles. This effect is especially prominent in patients who experience low LH before the start of GnRH antagonists. Keywords: Early pregnancy loss, GnRH antagonist, Luteinizing hormone, Menotropins, Ovulation induction

Background/Purpose An increasing number of human immunodeficiency virus-1 (HIV-l)-discordant couples in Taiwan have been seeking fertility help. We conducted the first clinical trial in Taiwan of assisted reproductive technology (ART) using sperm washing and viral load measurement. Methods From 2005 to 2009, we performed 22 ART cycles on 14 HIV-1-discordant couples. The sperm washing involved density gradient centrifugation followed by swim-up method. HIV-1 RNA was checked by real-time reverse transcription-polymerase chain reaction with a sensitivity of 40 copies/mL. In addition, we enrolled two other groups of ART recipients using frozen sperm to compare the clinical outcomes. Results There were five pregnancies in the fresh cycles (23.8%) of HIV-1-discordant couples and the cumulative pregnancy per couple was 42.9% (6/14). The data were comparable with normal controls and testicular sperm extraction/microscopic epididymal sperm aspiration groups. The nine babies and the 14 women in this study showed no seroconversion. Conclusion The preliminary data showed good ART results in HIV-1-discordant couples. Fertility services should not be withheld from individuals with HIV-1, although larger series are needed to reach conclusions about safety.

The serum lysophospholipase D activity and production of lysophosphatidic acid (LPA) increase in women with pregnancy. The effects of LPA on human placenta tissue remained unclear. We investigate the expression of LPA receptors and function of LPA in human first-trimester trophoblasts. Normal villous trophoblasts were obtained from termination of first-trimester gestation. We examined the expression of LPA receptors in primary culture of trophoblasts and the tissue. The effects of LPA on the expressions of chemokines of trophoblasts were examined using RT-PCR and enzyme immunoassay. We delineate signal pathways of LPA-inducing relevant chemokines in trophoblasts. The secretory chemokines were tested for angiogenic function using human endometrial microvascular endothelial cells and for immunological chemotaxis using decidual natural killer cells and THP-1 monocytes. The results revealed the expression of LPA1 receptors in trophoblast cells. LPA enhanced growth-regulated oncogene (GRO)-α, IL-8 and monocyte chemoattractant protein (MCP)-1 expressions in a time- and dose-dependent manner. Mechanistic dissection disclosed that LPA functioned mainly via the LPA1 receptor, Gi protein, various signal mediators of ERK, protein kinase C, p38, Akt, and c-Jun N-terminal kinase, and nuclear factor-κB pathways to secrete these chemokines. LPA-induced IL-8 protein secretion of trophoblasts enhanced permeability, migration, proliferation, and capillary tube formation of human endometrial microvascular endothelial cells. LPA-induced GRO-α and MCP-1 incited chemotaxis of natural killer cells and monocytes. We demonstrate that LPA mediates trophoblast cells to produce GRO-α, IL-8, and MCP-1 via LPA1 receptors and nuclear factor-κB-dependent signal pathways. Through LPA-induced chemokine production, human first-trimester trophoblast cells may regulate angiogenesis and innate immune system in early pregnancy.

Lysophosphatidic acid (LPA) is a pleiotropic phospholipid molecule involved in inflammation, angiogenesis, would healing,andcancerinvasion.WhereasserumlysophospholipaseD activity increases in women with pregnancy, the role of LPA inpregnancyremainsunclear.Weinvestigatedtheexpression of LPA receptors and function of LPA in endometrial stromal cells. Histologically normal endometrium was obtained from surgical specimens of women undergoing hysterectomy for leiomyoma. First-trimester decidua was obtained from women receiving elective termination of pregnancy. We examined the expressions of LPA1, LPA2, and LPA3 receptors in endometrial stromal cells. The effects of LPA on the expression of vascular endothelial growth factor, IL-6, and IL-8 were examined. Signal pathways of LPA were delineated. Functions of secretory angiogenic factors were tested using human endometrial microvascular endothelial cells. Immunoreactivity and mRNA of LPA1 receptors were identified in endometrial stromal cells. LPA enhanced IL-8 expression in a dose- and time-dependent manner, whereas vascular endothelial growth factor or IL-6 expression was not affected by LPA treatment. Mechanistic dissection disclosed that LPA functioned via the Gi protein, MAPK/p38 and nuclear factor-B pathway. LPA-induced IL-8 enhanced migration, permeability, capillary tube formation, and proliferation of human endometrial microvascular endothelial cells. Endometrial stromal cells express LPA1 receptors. Through the LPA1 receptor, LPA induces IL-8 expression via a nuclear factor-B-dependent signal pathway. These results could suggest that LPA may play a role in angiogenesis of endometrium and placenta through induction of IL-8 in endometrial stromal cells during pregnancy. (Endocrinology 149: 5888–5896, 2008)

Objective: This study tested the relationship between cellular immunity and the menstrual cycle in Taiwanese HIV-infected and normal women. Methods: From October 1997 to October 2001, 21 HIV-seropositive women and 30 controls were enrolled in this study. Blood was sampled for hormone profile (estradiol and progesterone) and immunophenotyping with flow cytometry during the follicular and luteal phases. Immunophenotyping included total blood cell count, lymphocyte count, CD4(superscript +) cells, CD8(superscript +) cells, and their activation markers, including CD25, CD69, HLA-DR, and CD38. Results: The proportion of CD8(superscript +) T cells increased during the follicular phase and activating antigens (HLADR and CD38) were elevated on CD8(superscript +) T cells of HIV-seropositive women. All these alterations seemed unrelated to the menstrual cycle. Conclusions: The CD8(superscript +) T cells were increased and activated in women with HIV infection but these alterations were not affected by the menstrual cycle. Therefore, sex hormones seem not to affect the course of HIV infection.

BACKGROUND: Uterine expression of leukemia inhibitory factor (LIF) is absolutely essential for mouse, and critical for human, embryo implantation. However LIF is not required for post-implantation development of mouse embryo. The objective of this study was to examine the role of LIF system in post-implantation stage of human pregnancy. METHODS: Tissues from 25 patients with anembryonic pregnancy (AP; blighted ovum) and 25 matched patients with normal pregnancy (NP) were collected. LIF and its receptor beta (LIF-Rbeta) expression in the decidua and chorionic villi were analyzed by semi-quantitative reverse transcription and polymerase chain reaction (RT-PCR), real-time quantitative PCR and immunohistochemical study. RESULTS: LIF mRNA levels were not different either between different tissues (decidua vs chorionic villi) or between different patients (NP vs AP). LIF-Rbeta mRNA levels were significantly higher in chorionic villi than in decidua but were not different between NP and AP. Immunohistochemical staining supported these findings and showed a predominate expression of LIF-Rbeta in the trophoblast cells. CONCLUSIONS: This study concluded that at early human post-implantation stage, LIF is produced from both decidua and chorionic villi and may exert its major action on trophoblasts. A baseline expression of LIF and LIF-Rbeta is probably needed for early pregnancy, but AP cannot be accounted for by the defective expression of either LIF or LIF-Rbeta in most circumstances.

Objective To compare the efficacy of coasting to prevent ovarian hyperstimulation syndrome (OHSS) with i.v. albumin and to determine if different timing of gonadotropin coasting would affect the IVF outcomes. Design Retrospective study. Setting University hospital–based IVF program. Patient(s) One hundred sixty-two women undergoing IVF treatment cycles who were considered to be at high risk for developing OHSS. Intervention(s) Gonadotropin administration was withheld in 60 patients: either coasting occurred before follicular maturation (early coasting subset, n=28) or follicular maturity was attained before coasting (late coasting subset, n=32). Outcomes were compared in 102 patients at risk for OHSS who received i.v. albumin on the day of oocyte retrieval. Main outcome measure(s) Incidence of OHSS and outcome parameters. Result(s) Although the fertilization rates and the incidence of OHSS did not differ significantly between the coasting and i.v. albumin groups, the mean number of oocytes retrieved, the pregnancy rates, and implantation rates were significantly lower in the coasting group. There is no statistical difference in the entire outcome examined and the incidence of OHSS between the early and late coasting subsets. Conclusion(s) Coasting is as effective as i.v. albumin in preventing OHSS in high-risk patients but yields inferior pregnancy rates. Early coasting is as successful as late coasting in preventing OHSS with similar IVF outcome.

Embryo freezing has been a successful practice, but oocyte cryopreservation formerly achieved poorer results. This was mainly due to low rates of survival, fertilization, and development. The major dissimilarities for oocytes to embryos are the character of the plasma membrane, the presence of cortical granules, at the metaphase of meiosis II with the spindle system. In addition, the oocytes must be fertilized by sperm at the appropriate time. To improve the survival rate, a refined slow freezing method with increased sucrose concentration would dehydrate oocytes more sufficiently. Vitrification is another approach to prevent ice crystal formation. Intracytoplasmic sperm injection is used to overcome possible zona hardening from the release of cortical granules. The microtubules of meiotic spindles are vulnerable to the thermal changes and would depolymerize. Cryopreserved oocytes exhibited serious disturbances of the microtubules immediately after thawing. Fertilization of oocytes with disorganized spindles could lead to chromosomal aneuploidy, digyny, and arrest of cleavage. After incubation, the microtubules would repolymerize in a time-dependent way. Normal fertilization and development of cryopreserved oocytes improved after appropriate incubation and timing of insemination, compatible with recovery of the spindles. With the improvement of survival, fertilization, and cleavage, oocyte cryopreservation would gain an imperative role.

Objective To analyze to what extent the parameters of ovarian functional reserve including female age and basal FSH levels will affect the results of ovarian hyperstimulation and IVF outcome. Design Retrospective cohort study. Setting University hospital infertility center. Patient(s) One thousand forty-five women undergoing their first cycle of IVF with ovarian stimulation after pituitary desensitization. Intervention(s) None. Main outcome measure(s) Cycle parameters, cancellation rate, implantation rate, and pregnancy rate. Result(s) Both increasing age and basal FSH were associated significantly with reduced numbers of oocytes collected, oocytes fertilized, and embryos transferred. The combined use of age and basal FSH significantly improves the predictive power for these parameters. Increasing age, but not basal FSH, was associated significantly with reduced implantation rate and pregnancy rate. Logistic regression analysis revealed that age, but not basal FSH, was an independent predictor of pregnancy rate. Neither age nor basal FSH had significant association with fertilization rate, miscarriage rate, or ectopic pregnancy rate. Conclusion(s) Both basal FSH and age contributed to the prediction of the quantitative ovarian reserve as reflected by the number of oocytes collected. However, age is a better predictor of pregnancy potential for women undergoing IVF.

Fertilization promoting peptide (FPP) and adenosine were demonstrated to be potential modulators of sperm capacitation in mammals. Both FPP and adenosine, by modulating the adenylate cyclase (AC)/cAMP signaling pathway, elicit similar biphasic responses in mammalian sperm (i.e., stimulating capacitation and inhibiting spontaneous acrosome loss). Pentoxifylline, an artificial sperm stimulant, is clinically used to enhance motility of sperm from infertile men. By inhibiting phosphodiesterase, pentoxifylline increases the intracellular cAMP level of sperm, and thus contributes to capacitation, hyperactivation, and acrosome reaction in animal studies. The effects of FPP, adenosine, and pentoxifylline on thawed human sperm are stressed. Chlortetracycline (CTC) fluorescence assessment revealed that none of the 3 reagents improved fertilization ability of post-thawed sperm. Motility studies with computer-aided sperm analyzer (CASA) showed significantly smaller STR (straight-line velocity) and LIN (linearity) in the FPP-treated group at 4 h of incubation por=005, significantly larger VCL in the adenosine-treated group por=.05, and significantly larger VCL (curvilinear velocity) and smaller LIN in the pentoxifylline-treated group por=.05. Significant decreases in percentage motility were also noted in both FPP and adenosine-treated groups por=05. It would appear that FPP potentiates fertilizing ability and prevents spontaneous acrosome loss, via regulating membrane-bound Na(+)-K(+) ATPase, and/or Ca(2+) ATPase, by keeping the sperm intracellular Ca(2+) concentration within the physiological range optimal for fertilization. adenosinefertilizationFPPpentoxifyllinespermthawing

Previous study demonstrated that IL-11 receptor alpha knockout female mice (IL-11Ralpha(-/-)) were phenotypically normal but infertile due to defective decidualization. However, the role of IL-11 signaling in human reproduction remains unclear. This study examined the expression of IL-11, IL-11Ralpha, and signal transduction factor glycoprotein 130 in different phases of endometrium (six in proliferative phase and four in secretory phase), and the decidua and villi of normal pregnancy (NP; n = 25) and anembryonic pregnancy (AP; n = 25) in the first trimester (gestational week 7-9). RT-PCR showed IL-11, IL-11Ralpha, and glycoprotein 130 mRNA expression in all samples, except the absence of IL-11 signal in the unstimulated MRC-5 cell and the proliferative phase endometrium. Real-time quantitative PCR showed that the relative level of IL-11Ralpha mRNA was not significantly different among proliferative phase endometrium (relative level; mean +/- SEM, 1.4 +/- 0.5), secretory phase endometrium (1.3 +/- 0.1), or decidua from NP or AP (1.7 +/- 0.3 and 1.9 +/- 0.4, respectively), but was significantly greater in chorionic villi either from NP or AP (7.6 +/- 1.3 and 10.6 +/- 1.9, respectively; both P0.05, compared with decidua or endometrium). No difference of IL-11Ralpha mRNA level was found between NP and AP (1.7 +/- 0.3 vs. 1.9 +/- 0.4 in deciduas; 7.6 +/- 1.3 vs. 10.6 +/- 1.9 in villi; both P0.05). In situ hybridization localized IL-11Ralpha mRNA expression in proliferative phase endometrium (stroma only), secretory phase endometrium (stroma and gland), decidua (stroma and gland), and villi (trophoblast and stroma). The staining intensities were not distinctly different between different groups of samples or between different cell types in a sample. No difference in IL-11Ralpha expression was found between NP and AP when either decidua or chorionic villi was analyzed. IL-11 mRNA level was not detected in the proliferative phase (relative level, 0.0 +/- 0.0), was barely detectable in the secretory phase (0.03 +/- 0.02), and was significantly increased in decidua (1.7 +/- 0.2 and 0.1 +/- 0.1, respectively, for NP and AP) and chorionic villi (13.0 +/- 2.2 and 0.2 +/- 0.1). In addition, IL-11 mRNA level was higher in NP than in AP both in decidua (1.7 +/- 0.2 vs. 0.1 +/- 0.1; P = 0.03) and in villi (13.0 +/- 2.2 vs. 0.2 +/- 0.1; P0.001). Immunohistochemistry study showed that IL-11 was nearly absent in endometrium in both phases, but clearly detectable in decidua and villi. Consistent with the results of quantitative PCR, the staining intensity was stronger in villi and decidua from NP than those from AP. The spatial and temporal changes in IL-11 and its receptor observed in this study suggest that IL-11 may be produced both by the embryo (predominantly) and the decidual cells and exerts its action on chorionic villi and decidua in an autocrine or paracrine manner. In the presence of a baseline level of IL-11Ralpha, IL-11 may subsequently regulate placentation and decidualization for the maintenance of a NP. The finding of decreased IL-11 expression in the absence of any change in IL-11Ralpha in AP suggests that defective expression of IL-11 but not IL-11Ralpha may account for certain cases of AP.

Background The aim of this study was to determine whether differences in cytokine concentrations, chemical composition, and murine embryogenesis in hydrosalpinx fluid could be observed between women with hydrosalpinx who achieved and did not achieve pregnancy. Methods Hydrosalpinx fluids were collected from 39 infertile women participating in an IVF-embryo transfer programme at a university teaching hospital. The fluids were analysed for concentrations of cytokine concentration (epidermal growth factor, interferon-gamma, leukaemia inhibitory factor, and tumour necrosis factor-alpha) and chemical composition. Mouse embryos were cultured in medium with various concentrations (0, 10, 50, 100%) of hydrosalpinx fluid. Results Logistic regression demonstrated that a murine blastocyst formation rate >or=53.3% in 50% hydrosalpinx fluid (OR = 16.6, 95% CI 2.4-116.1, P = 0.005) and patient age (OR = 0.778, 95% CI 0.61-0.99, P = 0.045) were independent predictors of IVF outcome. The diagnostic accuracy did not significantly improve when age and number of good quality embryos transferred was taken into account. The chemical composition and cytokine concentrations of hydrosalpinx fluid were not predictive of subsequent IVF outcome. Conclusions The mouse embryo assay of hydrosalpinx fluid might potentially serve as a predictor of subsequent IVF outcome in women with hydrosalpinx. This technique may be useful in selecting appropriate treatment options for patients with hydrosalpinx undergoing IVF treatment.

Mostin vitrofertilization (IVF) experts and infertility patients agree that the most ideal assisted reproductive technology (ART) outcome is to have a healthy, full-term singleton born. To this end, the most reliable policy is the single-embryo transfer (SET). However, unsatisfactory results in IVF may result from plenty of factors, in which aneuploidy associated with advanced maternal age is a major hurdle. Throughout the past few years, we have got a big leap in advancement of the genetic screening of embryos on aneuploidy, translocation, or mutations. This facilitates a higher success rate in IVF accompanied by the policy of elective SET (eSET). As the cost is lowering while the scale of genome characterization continues to be up over the recent years, the contemporary technologies on trophectoderm biopsy and freezing-thaw, comprehensive chromosome screening (CCS) with eSET appear to be getting more and more popular for modern IVF centers. Furthermore, evidence has showen that, by these avant-garde techniques (trophectoderm biopsy, vitrification, and CCS), older infertile women with the help of eSET may have an opportunity to increase the success of their live birth rates approaching those reported in younger infertility patients.

PROBLEM: To study the correlation of interleukin (IL)-10. IL-1 I leukemia inhibitory factor (LIF), placental growth factor (PIGF), and transforming growth factor (TGF)-β and outcome of human pregnancy. METHOD OF STUDY: We prospectively measured the serum levels of these cytokines in patients undergoing in vitro fertilization (IVF) programs. A total of 60 women (non-pregnant, n = 27; early abortions, n = 12; normal pregnancies, n = 21) were enrolled. RESULTS: There was no difference in the cytokines studied on DO and D14 among the three groups of women. The increase in PIGF from DO to D14 after human chorionic gonadotropin (hCG) injection was greater in pregnant women than in non-pregnant women; however, the difference did not reach significance (P = 0.068). The increase in IL-10 production from D14 to D21 was significant in women with successful pregnancies compared to women in the abortion group. CONCLUSIONS: This increase in IL-10 may be important in sustaining a normal pregnancy early after implantation.

Background To determine whether there is a factor (or factors) in the peritoneal fluid of endometriosis patients that impairs embryo growth and embryo implantation. Methods Growth and development of two-cell mouse embryos which were cultured in media with peritoneal fluid from women with or without endometriosis and interleukin-1-beta (IL-1beta), interleukin-6 (IL-6), and tumor necrosis factor-alpha (TNF-alpha) levels in conditioned media were measured. Results The blastocyst rate in the non-endometriosis group was 46.4 +/- 31.1%, and that of the endometriosis group was 54.6 +/- 28.7%. Logistic regression analysis using the criteria of blastocyst development in 454 embryos, showed that the peritoneal fluid from endometriosis could promote (p=0.015) but IL-6 could arrest embryo growth to blastocyst (p=0.025). IL-1beta and TNF-alpha levels had no significant effect on blastocyst formation. Conclusion Peritoneal fluid from women with endometriosis was not toxic to mouse embryo development. However, IL-6 in the peritoneal fluid deteriorated the growth and development of mouse embryos.

Vitrification of oocytes has been applied recently for humans, but remains elusive. The microtubules of oocytes are vulnerable to cryoprotectants and thermal changes. Using mouse oocytes, the effects of vitrification in open pulled straws (OPS) were investigated on survival, the meiotic spindle, and chromosomes and compared with conventional straws. Mature oocytes were allocated to four groups for exposure to cryoprotectants, vitrification in conventional straws, or vitrification in OPS. They were diluted in stepwise sucrose solutions. Oocytes without treatments were used as controls. The surviving oocytes were stained for meiotic spindles and chromosomes. After dilution, all of the oocytes exposed to cryoprotectants survived. Vitrification sometimes resulted in lysis so that survival using OPS (62%) was significantly (P < 0.05) smaller than that using conventional straws (81%). Oocytes exposed to cryoprotectants or vitrified exhibited serious disturbances of microtubules immediately post-dilution. After 1 h incubation, the microtubules could repolymerize so that the OPS group had significantly (P < 0.05) more normal spindles (78%) than did the conventional straw group (21%). The former also tended to have more compact chromosomes (87%) than did the latter (78%). OPS for vitrification of oocytes achieve more rapid cooling, warming, and dilution and so reduce spindle injury. However, the lower survival rate in OPS needs improvement.

Objective To examine the effect of vitrification with ethylene glycol (EG) for mature human oocytes in straws. Design Prospective, randomized, in vitro experiments. Setting Reproductive unit of a university hospital. Patient(s) Immature oocytes from 110 patients undergoing intracytoplasmic sperm injection (ICSI). Intervention(s) The immature oocytes were incubated to reach metaphase II (MII). The MII oocytes were treated with EG-based cryoprotectants and vitrified in straws. They were diluted in sucrose solutions, inseminated by ICSI, and cultured in vitro. Main outcome measure(s) Survival, fertilization, and embryo cleavage. Result(s) The survival rates were greater for oocytes pretreated with 1.5 M of EG (65% for 0 minute, 93% for 5 minutes, and 96% for 10 minutes). The oocytes vitrified in 60 and 90 seconds had a greater rate of fertilization than those vitrified in 120 seconds. There were no differences in survival and fertilization for vitrified oocytes diluted by three or four steps. The cleavage rates to the six- to eight-cell stage were comparable with controls. However, no blastocyst formation was observed in vitrified oocytes. Conclusion(s) Vitrification of human oocytes with EG in straws achieves a high rate of survival, fertilization, and early cleavage of embryos. Further studies should be conducted for the improvement of blastocyst formation.

Objective: To investigate the immunologic deviations of postmenopausal women before and after hormone replacement therapy (HRT). Design: Prospective study. Setting: University teaching hospital. Patient(s): Seventeen postmenopausal women (study group) and 17 women of reproductive age (control group). Intervention(s): Continuous usage of E 2 valerate 2 mg/d and medroxyprogesterone acetate 5 mg/d in postmenopausal women in the study group. Main Outcome Measure(s): Immunophenotyping with flow cytometry, cytokine production with and without mitogen stimulation of the peripheral mononuclear cells, and a natural killer (NK) cell cytotoxicity test against K562 target cells by the 51 Cr-release assay were performed in the control group and in the study group before, 1 month after, and 6 months after HRT. Result(s): NK cytotoxicity, interferon-γ production, and the T-cell subpopulation were significantly decreased, and the subpopulations of CD3 + CD25 + and CD3 + HLA-DR + were increased in the study group before HRT when compared with those in the control group. After HRT was given for 6 months, however, the NK cytotoxicity increased significantly in the postmenopausal women to a value similar to that of the control group. Conclusion(s): Women after menopause are prone to impaired immune responses. Nevertheless, some of the impairment can be restored after HRT.

PROBLEM Previously, we observed that follicular fluid obtained from patients with premature luteinization contained elevated interleukin-1 beta (IL-1 beta) levels. In this study. we aimed to examine the effects of luteinizing hormone (LH) on IL-1 beta expression and IL-1 beta on steroidogenesis in human granulosa-luteal cells. METHOD OF STUDY Human granulosa-luteal cells were obtained during oocyte retrieval. The cells were treated with either LH or IL-1 beta and subsequently were examined for the level of IL-1 beta transcript. The conditioned media were examined for IL-1 beta protein and steroid hormone levels. RESULTS LH (250-500 mIU/mL) up-regulated the expression of IL-1 beta mRNA (up to a 4-fold increase over control; P

The establishment of the human placenta in early pregnancy is characterized by the presence of large numbers of natural killer cells within the maternal decidua. These NK cells have an unusual phenotype, CD3- CD16- CD56(bright), distinguishing them from peripheral blood NK cells. They may control trophoblast migration and placentation. Using a panel of monoclonal antibodies to several members of the KIR family and flow cytometry, we found that KIRs are expressed on decidual NK cells. There is variation in both the percentage of cells expressing a particular receptor and the density of receptor expression between decidual NK cells from different individuals. In anembryonic pregnancy, the proportions of decidual NK cells with a particular KIRs (GL183 and EB6) decreased significantly when compared with normal pregnancy (p = 0.01 and 0.01, respectively), raising the possibility that these NK receptors may be involved in recognition of the allogeneic fetus by the mother at the implantation site. In the decidua, more CD4+ and CD8+ T cells expressed CD69 and HLA-DR than in blood, indicating that T cells are regionally activated during early pregnancy. When compared with normal pregnancy, decidual HLA-DR+CD4+CD3+, CD69+CD8+CD3+ and HLA-DR+CD8+CD3+ T lymphocytes are significantly increased in anembryonic pregnancy. The over-activation of decidual T cells during anembryonic pregnancy may thus contribute to the increased NK cytotoxicity activity.

Objective: To evaluate the efficiency and efficacy of a simplified two-pipette technique in comparison to the conventional three-pipette method; in the two-pipette method, a single, larger drilling/biopsy pipette is used to perform zona pellucida (ZP) drilling and blastomere aspiration for embryo biopsy. Design: A preclinical, prospective, randomized, in vitro experiment. Setting: The reproductive unit of a university teaching hospital. Patient(s): Ninety-five excess embryos at the two- to four-cell stage were obtained from 35 patients undergoing IVF. Intervention(s): At the six- to eight-cell stage, 88 embryos were allocated randomly to three groups: group I for the conventional method ( n = 29), group II for the simplified technique ( n = 30), and group III for controls ( n = 29). The embryos then were cultured in vitro. The retrieved blastomeres were fixed and examined with fluorescence in situ hybridization using X and Y probes simultaneously. Main Outcome Measure(s): Biopsy time, successful retrieval of a blastomere, fixation of the cell, signals developed from fluorescence in situ hybridization, and growth potential and hatching capacity of the biopsied embryos were evaluated. Result(s): The mean time (±SD) for biopsy of each embryo in group I (435 ± 137 seconds) was significantly longer than that in group II (126 ± 32 seconds). The success rates for obtaining an intact blastomere were not different between group I (93%) and group II (97%). The growth capacity to the blastocyst stage was similar among the three groups (34%, 37%, and 38%, respectively). However, the ZP-drilled and biopsied embryos of groups I and II had higher percentages of hatching (34% and 37%, respectively) and complete hatching (17% and 20%, respectively) than did those of group III (10% and 0, respectively). The blastomeres obtained by biopsy in groups I and II were equally fixed (90% vs. 90%, respectively) and shown in fluorescence in situ hybridization (79% vs. 80%, respectively). Conclusion(s): Compared with the conventional method, the simplified technique is more efficient and equally efficacious for blastomere biopsy in preimplantation genetic diagnosis.

To compare the effectiveness of various sperm preparation methods, we examined sperm concentration, percentage of progressive motility, recovery rate, and various motion characteristics in 32 semen samples after Percoll and IxaPrep preparations. Except for sperm concentration, which was much higher after IxaPrep preparation, no statistical differences existed between these two methods in terms of motile sperm concentration (MSC), recovery rate, mean curvilinear velocity (VCL), mean straight-line velocity (VSL) and mean amplitude of lateral head displacement (ALH). Among 22 samples that were allocated in a 37 degrees C, 5% CO2 incubator, the mean MSC after 3 hours was significantly higher following IxaPrep preparation than following Percoll preparation (63.2 x 10(6)/mL vs. 42.8 x 10(6)/mL, p.03). VCL and VSL were much higher after IxaPrep preparation than after Percoll preparation, both at 3 hours (p.01) and at 24 hours (p.03). ALH was also higher after IxaPrep preparation than after Percoll preparation, and it was statistically significant after 3 hours of incubation (p.03). We conclude that the IxaPrep gradient may be adopted as the preferred method to the Percoll gradient in preparing spermatozoa for assisted reproduction because of the spermatozoa's persistent and better motion activities after incubation.

Objective: To assess the effect of augmenting IVF with assisted hatching in the treatment of patients with repeated IVF failures. Design: Prospective randomized study. Setting: Division of Reproductive Endocrinology and Infertility of National Taiwan University Hospital. Patient(s): From July 1993 to February 1996, 49 patients with repeatedly failed IVF were treated with assisted hatching and were compared with 51 control subjects without assisted hatching. Intervention(s): Assisted hatching. Main Outcome Measure(s): Pregnancy rate and implantation rate per embryo after IVFET or IVF-tubal ET (TET) were measured. Result(s): The pregnancy rate (PR) in the assisted hatching group was found to be 36.7% compared with 19.6% in the control group, but the difference was not significant. When only patients receiving IVF-ET were considered, it was observed that the PR was significantly higher in the assisted hatching group than the control group (42.4% versus 16.1%). With IVF-TET however, the PR was found to be similar in both assisted hatching and control groups (25.0% and 25.0%, respectively). The rate of embryonic implantation in the IVF-ET patients was 11.0%, which was significantly higher than that of control embryos (3.7%). Conclusion(s): These results implied that IVF-ET, combined with assisted hatching, may improve the PR and implantation rate in patients with repeated IVF failures, but the same was not true in the case of IVF-TET.

Using different micropipettes for zona drilling and blastomere aspiration for embryo biopsy is prevalent at centers of preimplantation genetic diagnosis. The purpose of our study was to simplify the technique by using only one micropipette.In this animal model, ICR mouse embryos at the four-cell stage (n = 446) were randomly allocated into two groups; a biopsied group (n = 224) for blastomere aspiration and a control group (n = 222) without micromanipulation. We used a drilling/biopsy micropipette to drill a hole in the zona by expulsion of acidified Tyrode's solution and to aspirate the blastomere by gentle suction with the same micropipette and pull it out of the zona. One blastomere was biopsied from each embryo.In all, 222 (99.1%) intact blastomeres were successfully biopsied from 224 embryos. Only two blastomeres were damaged during aspiration. The capacity for blastocyst development (92.4 vs 93.7%) was not different between the two groups, but the percentages of embryos hatching (51.8 vs 18.0%) and hatched (29.9 vs 8.1%) were significantly higher in the biopsied group than in the control group.This simplified technique of embryo biopsy is safe and highly efficient for obtaining blastomeres for preimplantation genetic diagnosis and may also facilitate hatching of the blastocysts.

To clarify the effect of peritoneal fluids (PF) of women with various stages of endometriosis on sperm motility, we utilized computer-aides sperm analysis (CASA) to analyze sperm movement characteristics. PF was collected in the early follicular phase (days 4-8) of the menstrual cycle from 48 women undergoing diagnostic laparoscopy. Only sperm samples having normal sperm parameters were chosen for study. Swim-up separation was performed for 1 h at 37 degrees C. Sperm suspension was mixed at a ratio of 1:1 with each of the following four groups: group 1, human tubal fluid (HTF) with 10% fetal bovine serum (control), group 2, normal PF (n = 16); group 3, minimal or mild endometriosis PF (n = 16); and group 4, moderate or severe endometriosis PF (n = 16). The mixtures were analyzed at 0, 1, 3, 6, and 24 h of co-incubation using the CASA system. At the end of the 24 h incubation, Supravital staining of sperm was done to check the viability of sperm in each group. Only time (F = 126.6, p < .001) has a significant effect on sperm motion parameters. At 6 h, sperm velocity (mean curvilinear velocity and mean straight line velocity) of the PF groups was significantly greater than that of the control, but there was no significant difference between each PF group. At 24 h, the PF groups maintained 50% of initial sperm viability, compared with 13% of initial viability in the control group (p < .001). There was no adverse effect of PF in patients with endometriosis on sperm motion parameters.

The purpose of this study was to evaluate whether the combined treatment of growth hormone (GH) and gonadotropins can improve ovulation stimulation in previously poor responders. Twelve patients who, had suboptimal responses in previous in vitro fertilization cycle were enrolled. They underwent 1 cycle with gonadotropin-releasing hormone analogue (GnRH-a) and gonadotropins and another cycle with GnRH-a, gonadotropins, and GH. Serum gonadotropins, insulin-like growth factor-1 (IGF-1), and sex steroids, including estradiol (E2), progesterone (P4), testosterone, and androstenedione were measured on Day 2 and during ovulation induction. The serum IGF-1 level was higher in the GH cycle. There were no significant differences in the levels of the serum luteinizing hormone, E2, P4, testosterone, and androstenedione between the 2 cycles, so was IGF-1, E2 and P4 in follicular fluid. Co-treatment with GH did not improve the ovarian response. However, the GH cycles had better performance in terms of the number of oocytes fertilized and the pregnancy rate.

Objectives: Synchronous carcinomas of the endometrium and ovary may indicate either independently developing neoplasms or metastatic disease. The clinical implications and prognosis of these two categories are quite different. The objectives of this study were to identify and evaluate the empirical criteria and significant therapeutic implications. Method: The National Taiwan University Hospital Cancer Registry records and pathological reports from 1977 to 1994 were reviewed. Empirical criteria were used to identify synchronous primary cancers. Results: A total of 322 patients had endometrial cancer and 421 patients had ovarian cancer in our Cancer Registry records. Eleven patients had simultaneous cancer involvement of both the endometrium and ovary. Six cases fulfilled the criteria of synchronous primary carcinomas of the endometrium and ovary. Of these, five were alive and free of disease for 35–144 months (median 94.2 months). The disease-free survival rates between patients with synchronous primary and metastatic cancers of different histologic types showed a statistically significant difference (P = 0.013). No statistical significance was noted for different histologic types (P > 0.5). Conclusions: The empirical criteria used here were useful in identifying synchronous primary cancers of the endometrium and ovary. The favorable clinical outcome may relate to early detection of early-stage disease and low-grade malignancy with an indolent growth rate. Surgical management with or without adjuvant therapy has a satisfactory outcome in our experience.