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Objective: Oocyte (OC), embryo (EC), and ovarian tissue cryopreservation (OTC) are options for fertility preservation (FP) before going through gonadotoxic cancer treatment, or anticipated fertility decline in benign ovarian diseases, or for planned OC. The aim of this study is to report outcomes of FP in a single tertiary hospital in Korea., Methods: This is a retrospective study of OC, EC, and OTC cycles. All patients who visited or were referred to the infertility clinic at the Department of Obstetrics and Gynecology for the purpose of FP between 2010 and October 2021 were included., Results: A total of 564 controlled ovarian stimulation cycles were conducted in 416 women. Three hundred fifty-seven women underwent 494 OC cycles. Most patients were diagnosed with breast cancer (22.4%), followed by endometriomas (21.9%), and then by planned OC (20.7%). Cases of OC have increased over the years, peaking at 109 cycles in 2019 compared to one in 2010. Fifty-nine women underwent 70 EC cycles, and breast cancer (50.8%) was the most common indication. Repetitive OC and EC cycles were undergone in 92 and 9 women, respectively (mean number of repetition, 1.37 and 1.19 times in OC and EC, respectively), yielding a maximum number of 33 oocytes or 23 embryos being cryopreserved per patient. The utilization rate was 3.1% (11/357) in OC and 16.9% (10/59) in EC. Twenty-six women underwent OTC, and gynecologic cancer was the most common indication (9/26, 34.6%). One woman had the cryopreserved ovarian tissue retransplanted and successfully generated embryos., Conclusion: OC, EC, and OTC are possible options for preserving fertility, and these opportunities should be provided for women at risk of fertility decline or those who are eager to protect their future fertility. This is the first report on long-term FP outcomes in a single tertiary center in Korea. We expect that there will be more cases over the years and more women returning to use their gametes or embryos for pregnancy., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Choi, Hong, Kim, Kim, Lee and Suh.)

Humanity is in the midst of the coronavirus disease 2019 (COVID-19) pandemic, and vaccines-including mRNA vaccines-have been developed at an unprecedented speed. It is necessary to develop guidelines for vaccination for people undergoing treatment with assisted reproductive technology (ART) and for pregnancy-related situations based on the extant laboratory and clinical data. COVID-19 vaccines do not appear to adversely affect gametes, embryos, or implantation; therefore, active vaccination is recommended for women or men who are preparing for ART. The use of intravenous immunoglobulin G (IVIG) for the treatment of immune-related infertility is unlikely to impact the effectiveness of the vaccines, so COVID-19 vaccines can be administered around ART cycles in which IVIG is scheduled. Pregnant women have been proven to be at risk of severe maternal and neonatal complications from COVID-19. It does not appear that COVID-19 vaccines harm pregnant women or fetuses; instead, they have been observed to deliver antibodies against severe acute respiratory syndrome coronavirus 2 (SARSCoV-2) to the fetus. Accordingly, it is recommended that pregnant women receive COVID-19 vaccination. There is no rationale for adverse effects, or clinical cases of adverse reactions, in mothers or neonates after COVID-19 vaccination in lactating women. Instead, antibodies to SARS-CoV-2 can be delivered through breast milk. Therefore, breastfeeding mothers should consider vaccination. In summary, active administration of COVID-19 vaccines will help ensure the safe implementation of ART, pregnancy, and breastfeeding.

This study aimed to investigate the usability of blood markers for predicting controlled ovarian stimulation (COS) outcomes in patients with breast cancer undergoing fertility preservation (FP). In total, 91 patients with breast cancer who had undergone COS using a letrozole-combined gonadotropin-releasing hormone (GnRH) antagonist protocol before chemotherapy were enrolled retrospectively in a single tertiary hospital. FP outcomes were compared in terms of the mean platelet volume (MPV), MPV/platelet count (PC), neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR), and lymphocyte-to-monocyte ratio (LMR). The cutoff values for obtaining 10 or more mature oocytes as favorable prognoses were obtained for each parameter, and the COS outcomes were compared based on the cutoff values. The optimal cutoff levels for MPV and MPV/PC were 10.15 [sensitivity: 90.0%; specificity: 45.1%; AUC: 0.687; 95% CI (0.563, 0.810)] and 0.41 [sensitivity: 65.0%; specificity: 67.6%; AUC: 0.682; 95% CI (0.568, 0.796)], respectively. The oocyte numbers did not significantly differ with respect to the cutoff values of NLR, PLR, and LMR (p > 0.05). However, the total number of acquired and mature oocytes were significantly lower in the group with MPV<10.15 than in that with MPV≥10.15 (8.0 ± 5.1 vs. 12.6 ± 9.1, p=0.003; 4.0 ± 3.7 vs. 7.3 ± 6.3, p=0.002, respectively). Similarly, considering the cutoff of MPV/PC as 0.41, the low-MPV/PC group showed a significantly lower total oocyte yield than the high-MPV/PC group (9.5 ± 7.1 vs. 13.1 ± 9.1, p=0.048), whereas the number of mature oocytes showed similar patterns with no statistical significance (5.3 ± 5.4 vs. 7.3 ± 6.1, p=0.092). From logistic regression analysis, age, anti-Müllerian hormone (AMH) level, MPV, and MPV/PC≥0.41 were found to be significant factors for the acquisition of 10 or more MII oocytes (p=0.049, OR: 0.850; p<0.001, OR: 1.622; p=0.018, OR: 3.184; p=0.013, OR: 9.251, respectively). MPV or MPV/PC can be a reliable marker for predicting FP outcome in patients with breast cancer. Protocols to acquire more mature oocytes, such as the dual-trigger approach, could be recommended for patients with breast cancer with MPV<10.15. Furthermore, a higher dose of gonadotropins was considered to obtain more oocytes in patients with MPV/PC<0.41., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Hong, Kim, Lee and Suh.)

Purpose: To determine the optimal maturation method to increase the yield of mature oocytes, especially for cancer patients with fewer chances of fertility preservation (FP) before gonadotoxic therapy., Methods: A total of 373 cycles in 293 patients undergoing controlled ovarian stimulation (COS) for FP using a gonadotropin-releasing hormone (GnRH) antagonist protocol were enrolled. The control group ( n =  225) received 250 µg of recombinant human chorionic gonadotropin (rhCG) while the study group ( n =  148) received 250 µg of rhCG and 0.2 mg of triptorelin for triggering. Subgroup analyses were performed for stimulation cycles with diminished ovarian reserve (DOR; anti-Müllerian hormone (AMH) levels <1.1 ng/ml, n =  86), with endometrioma ( n =  104), or with breast cancer and endometrial cancer using 5 mg of letrozole during the COS cycles ( n =  84)., Results: There was no significant difference in the baseline characteristics or the number of total and mature oocytes between the two groups. Subgroup analyses for women with endometrioma or DOR showed similar results. However, the dual trigger group had a significantly higher number of mature oocytes than the rhCG trigger group in breast and endometrial cancer patients using letrozole during the COS cycles (6.9 ± 6.0 vs. 4.6 ± 3.6, p  = 0.034). The maturation rate was higher in the dual trigger group, although the difference was not statistically significant (59.3 ± 26.7 vs. 50.0 ± 28.0, p  = 0.124)., Conclusions: Dual triggering can be an efficient maturation method to maximize the yield of mature oocytes in breast or endometrial cancer patients using letrozole-combined GnRH antagonist protocol for FP., Competing Interests: None., (© 2022 The Authors. Reproductive Medicine and Biology published by John Wiley & Sons Australia, Ltd on behalf of Japan Society for Reproductive Medicine.)

Transplantation of ovary is one method of facilitating fertility preservation to increase the quality of life of cancer survivors. Immediately after transplantation, ovaries are under ischemic conditions owing to a lack of vascular anastomosis between the graft and host tissues. The transplanted ovaries can suffer damage because of lack of oxygen and nutrients, resulting in necrosis and dysfunction. In the technique proposed in this paper, the ovary is encapsulated with nitric oxide-releasing nanoparticles (NO-NPs) in fibrin hydrogels, which form a carrying matrix to prevent ischemic damage and accelerate angiogenesis. The low concentration of NO released from mPEG-PLGA nanoparticles elicits blood vessel formation, which allows transplanted ovaries in the subcutis to recover from the ischemic period. In experiments with mice, the NO-NPs/fibrin hydrogel improved the total number and quality of ovarian follicles after transplantation. The intra-ovarian vascular density was 4.78 folds higher for the NO-NPs/fibrin hydrogel groups compared to that for the nontreated groups. Finally, in vitro fertilization revealed a successful blastocyst formation rate for NO-NPs/fibrin hydrogel coated ovaries. Thus, NO-NPs/fibrin hydrogels can provide an appropriate milieu to promote angiogenesis and be considered as adjuvant surgery materials for fertility preservation., (© 2021 IOP Publishing Ltd.)

Endometrioma is known to reduce the ovarian reserve and the extent of the decrease is more severe when ovarian surgery is performed. Therefore, to prevent this decline in fertility, patients with endometrioma are considered candidates for preoperative fertility preservation (FP). In this study, we evaluate the efficacy of FP in women with endometrioma before planned ovarian surgery. A total of 95 cycles in 62 patients with endometrioma, undergoing controlled ovarian stimulation (COS) for FP using a gonadotropin-releasing hormone (GnRH) antagonist protocol before an expected ovarian surgery, were enrolled retrospectively. COS outcomes were compared according to endometrioma laterality. Additionally, first COS cycle outcomes in patients with endometrioma were compared with those in infertile patients, or in patients with a benign ovarian cyst using propensity score matching. When multiple COS cycles were performed, the results of cumulative cycles were analyzed. Embryo quality was worse in the bilateral endometrioma group. Compared with the infertile patient group, the patients with endometrioma had significantly lower Anti-Müllerian Hormone (AMH) and fewer numbers of oocytes retrieved (median, 3.3 vs. 1.2, p<0.001; 7.0 vs. 4.0, p=0.009, respectively). Compared with mature oocytes in infertile patients or patients with a benign cyst, mature oocytes were fewer in patients with endometrioma, but this was not statistically significant (median, 4.0 vs. 3.0, p=0.085; 5.5 vs. 3.0, p=0.052, respectively). The median value of the cumulative number of cryopreserved oocytes or embryos was 14.5 up to the fourth cycle compared to 3 up to the first cycle, with cumulative effect. Women with endometrioma should be counseled for FP before planned ovarian cystectomy. The number of cryopreserved oocytes or embryos can be increased by repeated cycles., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Hong, Lee, Kim, Lee and Suh.)

Study Objective: To analyze the obstetric and operative outcomes of 504 cases of single-port laparoscopic myomectomy (SPLM)., Design: Single-center retrospective study., Setting: A tertiary university hospital., Patients: A total of 502 patients (504 SPLM procedures) who underwent SPLM for symptom relief or growing myomas between October 2009 and April 2020., Interventions: Data on patient demographics, operative variables (estimated blood loss, hemoglobin decrease, operation time, perioperative complications, and postoperative hospital stay), and obstetric outcomes (the surgery-to-pregnancy interval and birth-related outcomes) were obtained from medical records and analyzed., Measurements and Main Results: The mean age of the patients was 40.6 ± 6.6 years. The patients had had an average of 2.3 ± 2.2 myomas removed; the largest myoma size was 6.8 ± 2.4 cm. The mean operation time, postoperative hemoglobin decrease, and postoperative hospital stay duration were 112.9 ± 45.3 minutes, 1.7 ± 1.1 g/dL, and 2.2 ± 1.4 days, respectively. The overall rate of postoperative complications was 7.7% (39/504), and the common complications were transfusions (16/504, 3.1%) or wound problems (15/504, 3.0%). Conversion to multiport or open myomectomy was required in 0.8% of the cases (4/504). A total of 376 women were of child-bearing age, and 56 attempted to become pregnant after surgery. The mean interval from surgery to pregnancy was 15.6 ± 12.2 months. The obstetric outcomes were pregnancy (42/56, 75.0%), live birth (39/56, 69.6%), and miscarriage (2/56, 3.6%). One pregnant woman was lost to follow-up. The 39 live births predominantly involved full-term delivery (36/39, 92.3%), mostly through cesarean section (36/39, 92.3%). No postpartum complications were reported. The 2 most common obstetric complications were preterm labor (7.6%) and gestational diabetes (5.1%)., Conclusion: SPLM seems to be an effective procedure with good operative and postoperative obstetric outcomes for women with myomas who require surgery and may wish to subsequently become pregnant., (Copyright © 2021 AAGL. Published by Elsevier Inc. All rights reserved.)

Objective: This study aimed to investigate sperm motility and its changes after preparation as predictors of pregnancy in intrauterine insemination (IUI) cycles., Methods: In total, 297 IUI cycles from January 2012 to December 2017 at a single tertiary hospital were retrospectively analyzed. Patient and cycle characteristics, and sperm motility characteristics before and after processing were compared according to clinical pregnancy or live birth as outcomes., Results: The overall clinical pregnancy rate per cycle was 14.5% (43/297) and the live birth rate was 10.4% (30/289). Patient and cycle characteristics were similar between pregnant and non-pregnant groups. Sperm motility after preparation and the total motile sperm count before and after processing were comparable in terms of pregnancy outcomes. Pre-preparation sperm motility was significantly higher in groups with clinical pregnancy and live birth than in cycles not resulting in pregnancy (71.4%±10.9% vs. 67.2%±11.7%, p=0.020 and 71.6%±12.6% vs. 67.3%±11.7%, p=0.030, respectively). The change in sperm motility after processing was significantly fewer in the non-pregnant cycles, both when the comparison was conducted by subtraction (post-pre) and division (post/pre). These relationships remained significant after adjusting for the female partner's age, anti-Müllerian hormone level, and number of pre-ovulatory follicles. According to a receiver operating characteristic curve analysis, an initial sperm motility of ≥72.5% was the optimal threshold value for predicting live birth after IUI., Conclusion: Initial sperm motility, rather than the motility of processed sperm or the degree of change after preparation, predicted live birth after IUI procedures.

Antifreeze proteins (AFPs) can inhibit the freezing of body fluid at subzero temperatures to promote the survival of various organisms living in polar regions. Type III AFPs are categorized into three subgroups, QAE1, QAE2, and SP isoforms, based on differences in their isoelectric points. We determined the thermal hysteresis (TH), ice recrystallization inhibition (IRI), and cryopreservation activity of three isoforms of the notched-fin eelpout AFP and their mutant constructs and characterized their structural and dynamic features using NMR. The QAE1 isoform is the most active among the three classes of III AFP isoforms, and the mutants of inactive QAE2 and SP isoforms, QAE2 ACT and SP ACT , displayed the full TH and IRI activities with resepect to QAE1 isoform. Cryopreservation studies using mouse ovarian tissue revealed that the QAE1 isoform and the active mutants, QAE2 ACT and SP ACT , more effectively preserved intact follicle morphology and prevented DNA double-strand break damage more efficiently than the inactive isoforms. It was also found that all active AFPs, QAE1, QAE2 ACT , and SP ACT , formed unique H-bonds with the first 3 10 helix, an interaction that plays an important role in the formation of anchored clathrate water networks for efficient binding to the primary prism and pyramidal planes of ice crystals, which was disrupted in the inactive isoforms. Our studies provide valuable insights into the molecular mechanism of the TH and IRI activity, as well as the cryopreservation efficiency, of type III AFPs., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2021 The Author(s).)

Purpose: Today, male and female adult and pediatric cancer patients, individuals transitioning between gender identities, and other individuals facing health extending but fertility limiting treatments can look forward to a fertile future. This is, in part, due to the work of members associated with the Oncofertility Consortium., Methods: The Oncofertility Consortium is an international, interdisciplinary initiative originally designed to explore the urgent unmet need associated with the reproductive future of cancer survivors. As the strategies for fertility management were invented, developed or applied, the individuals for who the program offered hope, similarly expanded. As a community of practice, Consortium participants share information in an open and rapid manner to addresses the complex health care and quality-of-life issues of cancer, transgender and other patients. To ensure that the organization remains contemporary to the needs of the community, the field designed a fully inclusive mechanism for strategic planning and here present the findings of this process., Results: This interprofessional network of medical specialists, scientists, and scholars in the law, medical ethics, religious studies and other disciplines associated with human interventions, explore the relationships between health, disease, survivorship, treatment, gender and reproductive longevity., Conclusion: The goals are to continually integrate the best science in the service of the needs of patients and build a community of care that is ready for the challenges of the field in the future.

Research-Question: What is the clinical usefulness of oocyte cryopreservation for fertility preservation in women with ovarian endometriosis?, Design: Clinical characteristics were retrospectively analysed in 34 women with endometrioma before a planned ovarian cystectomy. Ovarian stimulation outcomes were compared according to laterality. A one-to-one propensity score-matched analysis was conducted to compare ovarian stimulation outcomes of the first cycle in patients with endometrioma undergoing fertility preservation with those in infertile patients without endometrioma who underwent IVF treatment. The number of oocytes cryopreserved in repeated ovarian stimulation cycles was analysed., Results: The mean endometrioma size at diagnosis was 6.0 ± 2.5 cm. The mean age, serum anti-Mullerian hormone levels and number of oocytes cryopreserved were 30.7 ± 5.9 years, 1.85 ± 1.14 ng/ml, and 4.8 ± 3.2, respectively. The number of oocytes cryopreserved in bilateral endometrioma compared with unilateral endometrioma patients was 4.1 ± 2.9 versus 5.7 ± 3.4 (P = 0.600). In the propensity score-matched cohort (n = 22 per group), the number of oocytes retrieved was significantly lower in the patients with endometrioma undergoing fertility preservation compared with that in infertile patients without endometrioma (5.4 ± 3.8 versus 8.1 ± 4.8; P = 0.045). A total of 13 (38.2%) patients with endometrioma underwent repeated stimulation. The median (interquartile range) number of cryopreserved oocytes at the first and the second cycle were 3.0 (2.5-6.0) and 5.0 (2.5-7.5), respectively., Conclusions: Women with endometrioma should be counselled about oocyte cryopreservation for fertility preservation before surgery. The number of cryopreserved oocytes can be increased by repeated oocyte retrieval., (Copyright © 2020 Reproductive Healthcare Ltd. Published by Elsevier Ltd. All rights reserved.)

To establish a protocol of optimized three-dimensional (3D) culture of ovarian follicles, various biomaterials have been investigated with regard to their properties and functions on in vitro follicle growth. The present study aims to compare the new biomaterial, extracellular matrix-derived soft hydrogel (ES-hydrogel) and alginate, and evaluate the effects of biomaterials on further in vitro 3D culture growth of ovarian follicle and oocyte maturation. The isolated follicles from mouse ovaries were randomly divided into two-dimensional (2D) culture, alginate and ES-hydrogel, and just seeded on culture wells (2D culture) or encapsulated with alginate or ES-hydrogel (3D culture). Culture media from each group were collected on days 4, 8 and 10 or 11 for 17ß-oestradiol (E2) and progesterone (P4) measurement. On day 10 of in vitro culture, follicular survival and pseudo-antrum formation rate were examined, and oocyte maturation was induced by adding human chorionic gonadotropin and epidermal growth factor. After 17 h, ovulated mature oocytes collected and analyzed for oocyte diameter, normal spindle and chromosome alignment configuration, reactive oxygen species (ROS) level, and mitochondrial membrane potential level. To compare mechanical properties of two biomaterials, storage modulus was measured with the advanced rheometric expansion system. Our results showed that follicles cultured in ES-hydrogel, were significantly superior to those cultured 2D or alginate in the pseudo-antrum formation rate, cumulus-oocyte complexes (COCs) rate, MII oocyte rate, normal spindle rate, and E2 production. The ES-hydrogel and alginate groups were not significantly different in follicle survival rate, oocyte diameter, P4 production, ROS, and mitochondrial membrane potential levels. The storage modulus of ES-hydrogel was lower than that of alginate, suggesting that the improved follicular physiology and oocyte maturation in the ES-hydrogel group was due to better hormone exchange through a less stiff encapsulating material. This study shows that 3D culture system using ES-hydrogel effectively improve the outcome of in vitro ovarian follicle culture, supporting follicle morphology and growth and enhancing oocyte maturation. It means one of the most important factors for 3D culture of ovarian follicle was the selection of appropriate and effective biomaterial that can preserve the structure and morphology of ovarian follicle and facilitate nutrition and hormone exchange., (Copyright © 2019 Elsevier Inc. All rights reserved.)

This study aimed to analyze the effects of a new protocol with letrozole on the outcomes of in vitro fertilization (IVF) cycles in women with endometriosis. This retrospective cohort study was conducted for women diagnosed with endometriosis undergoing IVF from an infertility clinic. A new protocol, combination therapy with letrozole and gonadotropin, was used from August 2016 to January 2018 ('protocol 1', n  = 38). From March 2014 to July 2016, conventional IVF with gonadotropin was administered ('protocol 2', n  = 26). Age and ovarian reserve were comparable between the two groups. The patients who received protocol 1 resulted in a significantly lower peak estradiol level in IVF compared with those received protocol 2 (722 ± 1076 pg/mL versus 2168 ± 1521 pg/mL, p  < .001). The length of stimulation, the total dose of gonadotropin, number of oocytes retrieved, fertilization rates, and number of embryos obtained were similar between the two groups. The mean percentage of mature oocytes was lower (69.9 ± 23.7% versus 80.2 ± 21.0%, p  = .029) in patients with protocol 1. While maintaining low estrogen levels, the combination therapy with letrozole and gonadotropin produce similar oocyte and embryo yield to the conventional IVF protocol in women with endometriosis.

Background: It is still one of the unresolved issues if germinal vesicle stage (GV) oocytes can be successfully cryopreserved for fertility preservation and matured in vitro without damage after warming. Several studies have reported that the addition of cyclic adenosine monophosphate (cAMP) modulators to in vitro maturation (IVM) media improved the developmental potency of mature oocytes though vitrification itself provokes cAMP depletion. We evaluated whether the addition of cAMP modulators after GV oocytes retrieval before vitrification enhances maturation and developmental capability after warming of GV oocytes., Methods: Retrieved GV oocytes of mice were divided into cumulus-oocyte complexes (COCs) and denuded oocytes (DOs). Then, GV oocytes were cultured with or without dibutyryl-cAMP (dbcAMP, cAMP analog) and 3-isobutyl-l-methylxanthine (phosphodiesterase inhibitor) during the pre-vitrification period for 30 min., Results: One hour after warming, the ratio of oocytes that stayed in the intact GV stage was significantly higher in groups treated with cAMP modulators. After 18 h of IVM, the percentage of maturation was significantly higher in the COC group treated with dbcAMP. The expression of F-actin, which is involved in meiotic spindle migration and chromosomal translocation, is likewise increased in this group. However, there was no difference in chromosome and spindle organization integrity or developmental competence between the MII oocytes of all groups., Conclusions: Increasing the intracellular cAMP level before vitrification of the GV oocytes maintained the cell cycle arrest, and this process may facilitate oocyte maturation after IVM by preventing cryodamage and synchronizing maturation between nuclear and cytoplasmic components. The role of cumulus cells seems to be essential for this mechanism.

Recurrent implantation failure (RIF) is defined when pregnancy failure occurs after two consecutive in vitro fertilization-embryo transfers to the endometrium using at least four high-quality embryos in women. MicroRNAs are well-known function modulators and are involved in many diseases. Recently, studies on microRNA and recurrent pregnancy loss (RPL) have been actively carried out; however, single nucleotide polymorphisms of miRNA in RPL are not well known. Therefore, we set the aim of this study to identify whether polymorphisms in miRNAs that miR-27aA>G, miR-423C>A, miR-449bA>G, and miR-604A>G are risk factors for idiopathic recurrent implantation failure (RIF) in Korean women. Genotyping was assessed with a polymerase chain reaction-restriction fragment length polymorphism assay. We examined polymorphisms in four miRNA genes: miR-27aA>G, miR-423C>A, miR-449bA>G, and miR-604A>G. We found that the miR-27aA>G, miR-449bA>G, and miR-604A>G polymorphisms were significantly associated with a risk of RIF. In addition, the miR-27aA>G and miR-449bA>G polymorphisms were associated with the frequency of implantation failures. Specifically, the miR-449bAG+GG genotype was associated with RIF prevalence (total RIF: adjusted odd ratio [AOR] = 1.584, 95% CI = 1.008-2.490, P = 0.046; IF ≥ 3 group: AOR = 1.747, 95% CI = 1.088-2.803, P = 0.021; IF ≥ 4: AOR = 1.932, 95% CI = 1.122-3.327, P = 0.018). Based on these results, the miR-449b A>G may be a predisposing factor to RIF susceptibility. However, the mechanism underlying the function of miR-449b A>G in RIF remains to be determined and further studies are needed to improve understanding of the roles of miR-449b A>G, using a larger and more heterogeneous cohort.

Uterine arteriovenous malformation (AVM) is a vascular hamartoma of the myometrium that mostly results from uterine tissue damage. Herein, we report a case of uterine AVM managed successfully by hysteroscopy. The patient had an induced abortion and subsequent persistent vaginal spotting and irregular active vaginal bleeding. Ultrasonography showed a 3.5 cm × 2.9 cm heterogeneous lesion in the endometrial cavity with increased vascularity. Symptoms were monitored without uterine artery embolization to avoid complications that may affect a future pregnancy. However, 10 days later, she presented with active vaginal bleeding. Hysteroscopic endometrial mass excision was performed. Her postoperative hemoglobin level and vital signs were stable. Biopsy of the excised mass revealed AVM. Her postoperative vaginal bleeding decreased significantly, and outpatient ultrasonography 1 month later showed no abnormal findings. This case confirms the feasibility and safety of hysteroscopic management of uterine AVMs. The hysteroscopic technique should be prioritized for managing uterine AVMs., Competing Interests: There are no conflicts of interest

Objective: We systematically evaluated if there are any clinical differences in the safety and feasibility of single port laparoscopic myomectomy when compared to conventional laparoscopic myomectomy., Study Design: A meta-analysis and systematic review was conducted in accordance with the PRISMA statement. We searched different databases including Medline, Embase, Cochrane, and Google Scholar up to April 2018. Comparative studies as randomized controlled trials and prospective and retrospective cohort studies were included., Results: A meta-analysis was performed on the safety, feasibility, and potential benefits. Two randomized controlled trials and six cohort studies (2 prospective and 4 retrospective) were meta-analyzed to compare the surgical outcomes of 907 patients. Rates of minor complications were not different between the two groups [OR 1.33 (0.67, 2.63), I 2  = 0%]. Major complications requiring re-operation occurred in two cases in the single port group and in the conventional group respectively. There was no significant statistical difference between the groups. Conversion to laparotomy did not occur in either group, and the conversion rate from single port to the multi-port operation was 2.1% (6/288). The pain score one hour after operation was significantly lower in the single port group [MD -0.41 point (-0.63, -0.18), I 2  = 3.7%]. However, this difference was not observed at six and 24 h postoperatively. There was no difference in the operation time, estimated blood loss, or hemoglobin decrease between the two groups., Conclusion: A meta-analysis showed that single port laparoscopic myomectomy is comparable to conventional laparoscopic myomectomy in terms of safety and feasibility and more advantageous in terms of immediate postoperative pain. If performed based on the appropriate patient selection criteria including size and number of myoma, single port laparoscopic myomectomy can have similar surgical outcomes to those of conventional laparoscopic myomectomy. Further research and resources are required to identify whether single port laparoscopic myomectomy is more beneficial in terms of cosmetic results, patient satisfaction, and pregnancy outcomes., (Copyright © 2019 Elsevier B.V. All rights reserved.)

In vitro follicle growth (IVFG) is an emerging alternative option for fertility preservation in women instead of ovarian tissue cryopreservation and transplantation. To widen the application of this technique, follicle cryopreservation should be established prior to clinical use. In the present study, we tried to determine the optimal vitrification protocol of mouse ovarian follicle for in vitro culture and oocyte maturation by comparing four different compositions of cryoprotective agents (CPA). Secondary follicles were mechanically isolated from 2-week-old BDF-1 mice and randomly assigned to fresh control and four different groups by the composition of CPAs (ES, EDS, EFS and EPS groups; E: ethylene glycol, D: dimethyl sulfoxide, S: sucrose, F: ficoll, P: 1,2-propanediol (PROH)). After vitrification and warming procedures, the follicles were cultured in vitro for 10 days and then treated with human chorionic gonadotropin and epidermal growth factor to induce oocyte maturation. Fourteen to 16 h later, oocyte maturation and quality were assessed. Follicle viability was evaluated by Calcein-AM/ethidium homodimer-1 staining immediately after warming, and their survival and diameters were measured during follicle culture periods. Antral cavity formation was observed at the end of the culture period (on the 10th day of culture). Following oocyte maturation, its maturational ability and meiotic spindle formation were assessed to evaluate their competence. There was no significant difference in viability after warming among the vitrification groups. From the 8th day of culture, the survival rate of ES and EDS were significantly higher than those of other vitrification groups (EPS and EFS). The follicle diameter was largest in the fresh-control group from the 6th day, while smallest in the EFS with statistical significance. On the 10th day of culture, the antral-cavity formation rate of EDS was comparable to that of the fresh control group. However, the oocyte maturation was significantly decreased in all four vitrification groups when compared with control group; especially, the EFS showed a more marked reduction in the oocyte maturation. There were no significant differences in meiotic spindle formation among all of those groups. Our results suggest that EDS combination for mouse follicle vitrification are the most effective vitrification protocols for mouse follicle and evaluated by an in vitro culture and oocyte maturation after warming., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Study Objective: To compare operative outcomes of single-port laparoscopic myomectomy (SP-LM) vs conventional laparoscopic myomectomy (CLM), including subjective and objective cosmetic aspects., Design: Prospective randomized controlled trial (Canadian Task Force classification I)., Setting: University hospital., Patients: Women with uterine myoma scheduled for laparoscopic myomectomy., Interventions: Sixty-six women were assigned at random to either the SP-LM or CLM group. Surgical outcomes, including patient and observer scar assessments, were evaluated between the groups according to the intention-to-treat principle., Measurements and Main Results: There were no significant differences in demographic characteristics and properties of myomectomy between the groups. There also were no differences in surgical outcomes, such as operation time, estimated blood loss, and complications, between the 2 groups. The mean total score of the Observer Scar Assessment Scale was lower in the SP-LM group at 1 week (13.0 ± 3.2 vs 18.3 ± 4.8; p < .001) and 8 weeks (9.9 ± 3.2 vs 14.3 ± 3.8; p < .001) after discharge. Similar results were obtained for the Patient Scar Assessment Scale at 1 week (11.6 ± 7.2 vs 18.5 ± 12.8; p = .024) and 8 weeks (9.5 ± 6.0 vs 18.8 ± 9.1; p < .001) after discharge. Postoperative pain and analgesic consumption did not differ between the groups, except in patient-controlled analgesia consumption at 6 hours after operation, which was lower in the SP-LM group (12.7 ± 6.3 mL vs 16.4 ± 6.2 mL; p = .039). Operative outcomes were similar in the 2 groups., Conclusion: SP-LM is associated with more favorable cosmetic outcomes and better patient satisfaction compared with CLM. There were no differences in operative outcomes and complications between the 2 modalities., (Copyright © 2017 American Association of Gynecologic Laparoscopists. Published by Elsevier Inc. All rights reserved.)

Success in cryopreservation of ovarian tissue (OT) in animal models has led to develop efficient cryo-technologies for human ovarian tissue. In this chapter, cryopreservation protocols developed for animal experiments are described.

In recent years, supplementation of antioxidants and antifreeze proteins during cryopreservation/vitrification has significantly improved the survival and function of oocytes and ovarian tissues (OT) in animal models. In this chapter, the experimental protocols for the use of antioxidants and antifreeze proteins in cryopreservation/vitrification are described.

Objective: Uterine contraction induced by the embryo transfer (ET) process has an adverse effect on embryo implantation. The aim of this study was to determine the effect of oxytocin antagonist supplementation on the day of ET on in vitro fertilization outcomes via a meta-analysis., Methods: We performed a meta-analysis of randomized controlled trials (RCTs). Four online databases (Embase, Medline, PubMed, and Cochrane Library) were searched through May 2015 for RCTs that investigated oxytocin antagonist supplementation on the day of ET. Studies were selected according to predefined inclusion criteria and meta-analyzed using RevMan 5.3. Only RCTs were included in this study. The main outcome measures were the clinical pregnancy rate, the implantation rate, and the miscarriage rate., Results: A total of 123 studies were reviewed and assessed for eligibility. Three RCTs, which included 1,020 patients, met the selection criteria. The implantation rate was significantly better in patients who underwent oxytocin antagonist infusion (19.8%) than in the control group (11.3%) (n=681; odds ratio [OR], 1.92; 95% confidence interval [CI], 1.25-2.96). No significant difference was found between the two groups in the clinical pregnancy rate (n=1,020; OR, 1.57; 95% CI, 0.92-2.67) or the miscarriage rate (n=456; OR, 0.76; 95% CI, 0.44-1.33)., Conclusion: The results of this meta-analysis of the currently available literature suggest that the administration of an oxytocin antagonist on the day of ET improves the implantation rate but not the clinical pregnancy rate or miscarriage rate. Additional, large-scale, prospective, randomized studies are necessary to confirm these findings., Competing Interests: No potential conflict of interest relevant to this article was reported.

Objective: Most interventions aimed at reducing bleeding during myomectomy lack sufficient evidence regarding their effectiveness. Recently, it was reported that intraoperative ascorbic acid administration effectively reduced blood loss during abdominal myomectomy. Therefore, this study aimed to investigate whether intravenous ascorbic acid infusion would affect intraoperative blood loss in women undergoing laparoscopic myomectomy., Study Design: A randomized, double-blind, parallel-group, placebo-controlled trial including 50 women undergoing laparoscopic myomectomy was conducted. Women with ≤4 myomas, ≤9cm in maximum diameter were eligible. The study:control group ratio was 1:1. Starting 30minutes before anesthesia, 2g of ascorbic acid or a placebo were administered for 2hours intraoperatively. Intraoperative blood loss, the primary endpoint, was calculated as the difference between the volume of fluids acquired from suction and that used for irrigation of the abdominal cavity during surgery using constant values., Results: Among the 50 randomized women, 1 and 3 in the study and control groups, respectively, were excluded due to withdrawal of consent, cancelation of surgery, or non-measurement of the primary endpoint. The baseline and operative characteristics were similar between the study and control groups, as was the intraoperative blood loss (193±204mL vs. 159±193mL, P=0.52). In addition, the operating time (95±29min vs. 110±52min; P=0.50) and decrease in hemoglobin level after surgery (1.9±1.31g/dL vs. 1.4±1.4g/dL; P=0.24) were similar between the study and control groups., Conclusions: Intravenous ascorbic acid infusion did not reduce intraoperative blood loss in women undergoing laparoscopic myomectomy., Clinical Trial Registration: ClinicalTrials.gov, www.clinicaltrials.gov, NCT01715597., (Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.)

Study Question: Are mRNAs for specific aquaporins (AQPs) expressed in human luteinized granulosa cells (GCs) and are their expression levels correlated with in vitro fertilization (IVF) outcomes?, Summary Answer: The mRNAs of AQP1-7, 9, 11, and 12 were expressed in human luteinized GCs; the level of AQP1 mRNA was negatively associated with retrieved oocyte number and the level of AQP7 mRNA was positively associated with fertilization rate., What Is Known Already: mRNAs of AQP1-4 and AQP9 have been detected in human GCs., Study Design, Size, Duration: Prospective observational study involving 111 women undergoing a stimulated IVF cycle., Participants/materials, Setting, Methods: In a preliminary experiment on 27 women, the mRNA expression of AQP0-12 in GCs was explored by RT-PCR. In the main experiment, luteinized GCs were obtained from 111 women at the time of oocyte retrieval of whom 102 had an embryo replacement. Real-time quantitative RT-PCR (qRT-PCR) was performed to quantify the mRNA level of AQP1-7, 9, 11, and 12. The mRNA for luteinizing hormone receptor (LHR) and steroidogenic acute regulatory protein (StAR) were also quantified by qRT-PCR., Main Results and the Role of Chance: mRNAs for AQP0, 8 and 10 were not detected in the preliminary experiment. In samples from 111 women, retrieved oocyte number was negatively associated with the mRNA levels of AQP1, 4, 6, and 11 and LHR (r = -0.311, r = -0.233, r = -0.203, r = -0.194, and r = -0.202, respectively, P < 0.05 for each), however, after adjustment for woman's age and serum anti-Müllerian hormone (AMH) levels, only correlation with AQP1 was found (r = -0.299, P < 0.05). BMI was negatively associated (after adjustment for age) with the mRNA level of AQP7 (r = -0.259, P < 0.05). Fertilization rate was positively associated with the mRNA level of AQP7 (r = 0.269, P < 0.05). The number or quality of embryos or clinical pregnancy was not associated with the mRNA levels of any of ten AQP subtypes. The mRNA levels for the ten AQP subtypes were correlated positively with LHR expression but negatively with StAR expression. Amongst high responders (oocyte number ≥14), the mRNA levels of AQP11 (1.4 ± 0.7 versus 1.7 ± 0.6) and LHR (1.3 ± 0.7 versus 1.7 ± 0.7) were significantly lower in the group with PCOS than in the non-PCOS group., Limitations, Reasons for Caution: A relative small number of subjects in PCOS group is the main limitation of our study. P-values were not corrected for multiple comparisons., Wider Implication of the Findings: AQP1 may be one of the factors that modulate individual ovarian response to exogenous gonadotrophin. The mRNA level of AQP7 was positively associated with fertilization rate, which is a surrogate marker of oocyte competence, thus expression of AQP7 could be a marker for adequate folliculogenesis and healthy oocytes., Study Funding/competing Interests: This work was supported by grant no. A120043 from the Korea Health Care Technology R&D Project, Ministry of Health and Welfare, Korea. None of the authors has any conflict of interest to declare., (© The Author 2016. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Aim: Few studies have reported that human papillomavirus (HPV) tests using menstrual blood (MB) may be a convenient and effective screening modality for cervical cancer. Therefore, we aimed to investigate the efficacy of detecting high-risk (HR)-HPV from MB in women with high-grade squamous intraepithelial lesions or HR-HPV infections dependent on menstrual days., Materials and Methods: In this prospective exploratory pilot study, a clinical trial was conducted in 19 women. On enrollment into the study, gynecologists collected cervical cells. On the first and second day of menstruation, MB was self-collected by patients using a sanitary pad with a filter. The distribution of HPVs from MB and the accuracy of menstrual HR-HPV tests were evaluated using HPV genotyping. The agreement rate of detecting HR-HPVs using cervical and MB samples was also investigated., Results: The sensitivity, specificity, positive and negative predictive values of the MB HR-HPV test for detecting cervical intraepithelial neoplasia (CIN) 3 or worse were 87.5%, 45.5%, 53.8%, and 83.3%, respectively, during both menstrual cycle day (MCD) 1 and 2 and MCD 1 only; and 62.5%, 27.3%, 38.5%, and 50.0%, respectively, during MCD 2 only. For CIN 3 or worse, the agreement rate between positive cervical and MB HR-HPV test results was 87.5% during MCD 1 and 62.5% during MCD 2., Conclusions: We demonstrated the possibility of using the MB HPV test as a screening modality for cervical cancer., (© 2015 Japan Society of Obstetrics and Gynecology.)

Purpose: Reducing the ischemic damage from free radicals that is inflicted on ovarian tissue is critical for successful ovarian tissue transplantation. Polyethylene glycol-superoxide dismutase (PEG-SOD) is mimetic of superoxide dismutase (SOD) and powerful free radical scavenger acts by reducing superoxide anions. The objective of study was to evaluate effects of PEG-SOD on mouse ovarian tissues in in vitro culture and in autotransplantation., Methods: Ovaries were collected and randomly divided into four groups that received different doses of PEG-SOD. To assess effects of PEG-SOD on in vitro cultures, four different doses of PEG-SOD were applied to in vitro culture media during in vitro culturing following ovarian tissue vitrification and warming. To evaluate effects of PEG-SOD on ovarian tissue transplantation, four different doses of PEG-SOD were applied for 2, 7, and 21 days to mice following vitrified-warmed mouse ovarian tissue autotransplantation., Results: The percentage of primordial follicles was maintained at the highest dose of PEG-SOD for 2 h in vitro, and there was a significant decrease in the percentage of apoptotic follicles at 2 h, but not at later time points. The highest dose of PEG-SOD also maintained primordial, primary, and secondary follicles 2 days post-transplantation, but only primordial follicles were maintained up to 21 days after transplantation., Conclusions: PEG-SOD is protective mainly toward primordial follicles only for a short interval in vitro, presumably via antioxidant effects. PEG-SOD may be a promising additive for preserving ovarian tissue integrity, at least for primordial follicles, up to 21 days post-transplantation.

Study Question: Can antifreeze proteins (AFPs) from three different sources improve the efficacy of mouse oocyte vitrification?, Summary Answer: Treatment with AFPs can improve both murine oocyte quality and embryo development, and reduce reactive oxygen species (ROS) production in vitrified-warmed oocytes., What Is Known Already: A previous study discovered that vitrification of immature oocytes and 2-cell stage embryos of mice augmented with antifreeze glycoproteins at 40 mg/ml dramatically improved the morphological integrity of the samples, suggesting that AFPs have the ability to inhibit ice formation and stabilize the plasma membrane., Study Design, Size, Duration: Metaphase II oocytes were obtained from 4-week-old BD-F1 mice. AFPs from bacteria (Flavobacterium frigoris ice-binding protein (FfIBP)), yeast (Glaciozyma sp. ice-binding protein (LeIBP)) and fish (Type III AFP) were added to the vitrification and warming solutions individually. Survival and development, meiotic spindle organization, intracellular ROS, mitochondrial activity, DNA double-strand breaks (DSBs) and repair of damaged DNA were analyzed., Participants/materials, Setting, Methods: Vitrification of oocytes was performed with the CryoTop (equilibration solution: 7.5% ethylene glycol (EG) and 7.5% 1,2-propandiol (PROH) for 5 min; vitrification solution: 15% EG, 15% PROH and 0.5 M sucrose for 1 min). Warming was performed in three steps with decreasing concentrations of sucrose (1.0, 0.5 and 0.25 M sucrose)., Main Results and the Role of Chance: AFP treatment can improve murine oocyte quality and embryo development. Survival rates, cleavage rates and blastocyst rates (blastocyst per cleaved and per survived oocytes) of oocytes in AFP-treated groups were significantly higher than those in the control group [75.0, 89.0, 90.0 and 85.0% for survival rate (P = 0.012); 58.7, 89.0, 87.8 and 81.2% for cleavage rate (P = 0.003); 52.3, 87.7, 78.5 and 76.8% for blastocyst per cleaved oocytes (P < 0.01); 30.7, 78.0, 68.9 and 62.4% for blastocyst per survived oocytes (P < 0.01) in control, FfIBP, LeIBP and Type III AFP-treated groups, respectively]. The mean (±SD) number of apoptotic blastomeres per blastocyst was significantly lower in AFP-treated groups than in the control group (9.1 ± 1.0, 2.0 ± 1.7, 2.3 ± 1.2 and 2.7 ± 2.4 in control, FfIBP, LeIBP and Type III AFP-treated groups, respectively, P = 0.040). FfIBP treatment was the most effective in maintaining normal meiotic spindle organization and chromosome alignment (52.0, 92.0, 80.0 and 83.0% in control, FfIBP, LeIBP and Type III AFP-treated groups, respectively, P < 0.01). Intracellular ROS levels (mean ± SD) significantly decreased in the AFP-treated groups (17.0 ± 11.2, 8.4 ± 8.2, 10.3 ± 6.4 and 11.6 ± 12.3 in control, FfIBP, LeIBP and Type III AFP-treated groups, respectively, P < 0.01), and the FfIBP and LeIBP groups had significantly lower DNA DSBs, compared with controls (65.2, 30.8, 44.4 and 55.8% in control, FfIBP, LeIBP and Type III AFP-treated groups, respectively, P < 0.01)., Limitations, Reasons for Caution: The origins of FfIBP and LeIBP were bacteria and yeast, respectively. Therefore, treatment of human oocytes and embryos with these AFPs should be tested before clinical application., Wider Implications of the Findings: After further research, AFPs can potentially be applied to human oocyte cryopreservation to improve the efficacy of vitrification., Study Funding/competing Interests: This study was supported by a grant of the Korea Healthcare technology R&D Project, Ministry of Health & Welfare, Republic of Korea (HI12C0055). The authors have no conflict of interest to declare., (© The Author 2015. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Study Objective: To compare the operative outcomes of patients undergoing either single-port or multiport laparoscopic hysterectomy (LH)., Methods: Two hundred fifty-six women scheduled for LH for symptomatic myoma and/or adenomyosis from 8 tertiary teaching hospitals were randomized to single-port or multiport groups. Primary outcome was conversion and/or complication proportion of the planned procedure to determine whether the success proportion of the single-port approach was not inferior to that of the multiport approach. Secondary outcomes were postoperative pain and operative scar., Results: Demographic parameters including age, body mass index, parity, and history of vaginal and cesarean delivery were comparable between the 2 groups. The primary outcome of a combined conversion and/or complication rate was similar between the single-port and multiport groups at 8% and 10.3%, respectively. Conversions were similar between the groups with 4% of single-port cases and .8% of multiport cases. Transfusions were the most frequent complication required in 4.0% of single-port cases and 7.9% of multiport cases, with no difference between the groups. Concerning secondary outcomes, postoperative pain score and patient and observer scar assessment were not different between the 2 groups. Although not a specific outcome measure, there was no difference between the groups in blood loss, operative time, and postoperative hospital stay., Conclusion: Single-port LH is not inferior to multiport LH in terms of conversion and/or complications rates, including transfusion. However, the single-port approach did not have any advantage over multiport LH with regard to pain or cosmetic outcomes. These findings were demonstrated by multi-institutional surgeons in Korea., (Copyright © 2015 AAGL. Published by Elsevier Inc. All rights reserved.)

Study Objective: To identify learning curves for single-port laparoscopic myomectomy (SPLM) and evaluate surgical outcomes according to the sequence of operation., Design: A retrospective study., Setting: A university-based hospital (Canadian Task Force classification II-2)., Patients: The medical records from 205 patients who had undergone SPLM from October 2009 to May 2013 were reviewed. Because the myomectomy time was significantly affected by the size and number of myomas removed by SPLM, cases in which 2 or more of the myomas removed were >7 cm in diameter were excluded. Furthermore, cases involving additional operations performed simultaneously (e.g., ovarian or hysteroscopic surgery) were also excluded. A total of 161 cases of SPLM were included., Interventions: None., Measurements and Main Results: We assessed the SPLM learning curve via a graph based on operation time versus sequence of cases. Patients were chronologically arranged according to their surgery dates and were then placed into 1 of 4 groups according to their operation sequence. SPLM was completed successfully in 160 of 161 cases (99.4%). One case was converted to multiport surgery. Basal characteristics of the patients between the 4 groups did not differ. The median operation times for the 4 groups were 112.0, 92.8, 83.7, and 90.0 minutes, respectively. Operation time decreased significantly in the second, third, and fourth groups compared with that in the first group (p < .001). Proficiency, which is the point at which the slope of the learning curve became less steep, was evident after about 45 operations., Conclusion: Results from the current study suggested that proficiency for SPLM was achieved after about 45 operations. Additionally, operation time decreased with experience without an increase in complication rate., (Copyright © 2015 AAGL. Published by Elsevier Inc. All rights reserved.)

Necrostatin 1 (Nec1) is widely used in disease models to examine the contribution of receptor-interacting protein kinase 1 in cell death. The biological actions of Nec1 are blocking necrotic cell death. The purpose of this study was to investigate whether adding Nec1 into in vitro maturation (IVM) media, followed by vitrification procedures, could enhance the survival and developmental competency of oocytes. Germinal vesicle oocytes were matured in IVM medium containing 2 different doses of Nec1 (0.5 and 1 μmol/L). After IVM, the oocytes were vitrified using a 2-step exposure to equilibrium and vitrification solutions. After warming, the rates of survival, fertilization, embryonic development up to blastocyst in vitro, morphology of spindle and chromosome, membrane integrity, mitochondria integrity, and several gene expressions were evaluated. The survival and developmental competency of oocytes were higher in the 1 μmol/L Nec1-treated group than control. The proportion with intact spindles/chromosomes and stable membranes was similar in all the groups. The mitochondrial integrity of all Nec1-treated groups showed a higher score with strong staining. The 1 μmol/L Nec1 showed significantly increased expressions of Mad2, Gdf9, and Bcl2. The Cirp level had a tendency to be downregulated in the 0.5 µmol/L Nec1 but upregulated in the 1 μmol/L Nec1, compared with the control. The Mtgenome expressions were significantly decreased in both Nec1 groups. The supplementation of 1 μmol/L Nec1 into the IVM medium could be beneficial for the survival and development of immature oocytes after vitrification., (© The Author(s) 2014.)

Many studies have shown that ischemic injuries during the transplantation process were more detrimental than cryoinjuries for follicle survival and death, and it has been reported that transplantation sites can affect the outcomes of grafted ovarian tissue (OT). The purpose of this study was to assess the impact of different OT transplantation sites on follicular integrity and function of OT grafts. B6D2F1 mice were randomly assigned to control (sham) and four experimental groups according to transplantation sites (back muscle [BM], fat pad [FP], kidney capsule [KC], and subcutaneous [SC]). The ovaries from four groups were autotransplanted to each site. The OT recovery ratios on Days 2, 7, and 21 were significantly decreased in the FP group. The mean numbers of follicles were significantly lower in all the grafting groups compared with the sham group, except in the KC group on Days 7 and 21 and the BM group on Day 21. On Day 2, all the experimental groups showed low intact (G1) follicle ratio when compared with the sham group; however, the BM, KC, and FP groups recovered their morphologic integrity on Day 7, and only the SC group presented a significant decrease in G1 follicle ratios. On Day 21, the G1 follicle ratios of the FP and KC groups were greater than the sham control group. The proportion of apoptotic follicles of the four OT graft groups was higher than in the sham group on Day 2, followed by a significant decrease in the KC group and an increase in the SC group on Day 7. The serum follicle-stimulating hormone levels were significantly increased in all grafting groups on Day 2. On Day 7, only the SC group showed the high follicle-stimulating hormone level compared with the other groups. The mean numbers of oocytes from OT grafts were the highest in the KC group, except in the control group, and the lowest in the SC group. The ratios of mature oocytes were also significantly greater in the sham and KC groups. However, the ratios of normal spindle did not differ among the five groups. In conclusion, the KC was the optimal site for OT transplantation in this murine model, whereas the SC site was unfavorable for this procedure. In this study, we confirmed that the different grafting sites influenced the outcomes of transplantation., (Copyright © 2015 Elsevier Inc. All rights reserved.)

After the ovarian tissue (OT) transplantation, the ischemia-reperfusion injury causes depletion and apoptosis of follicle. Recent reports stated that simvastatin reduces ischemic damage. Therefore, we used the mouse whole ovarian vitrification and autotransplantation models to investigate the effects of simvastatin. Five-week-old B6D2F1 mice were randomly divided into four groups. Three groups were given simvastatin orally (5 mg/kg) before ovariectomy, either 2 hours before (2H Tx) or once a day for 3 or 7 days. The control group was given saline 2 hours before ovariectomy. All ovaries were cryopreserved by vitrification, held in liquid nitrogen for 1 week before being warmed, and autotransplanted. The grafts were collected for analysis on 2, 7, or 21 days after transplantation. Ovarian follicle morphology and apoptosis were assessed by hematoxylin and eosin staining and terminal deoxynucleotidyl transferase dUTP nick end labeling assay. Vessel integrity in ovary was evaluated by immunohistochemistry using anti-CD31 antibody. Serum FSH level was measured to estimate the transplanted ovarian reserve. The proportion of morphologically normal (G1) follicles at 7 and 21 days and the percentage of CD31 (+) tissue at 21 days was significantly higher in the 2H Tx group than that in the control group. In addition, the 2H Tx group showed a significantly increased intact primordial follicle ratio at 2 and 21 days after OT transplantation. Administration of simvastatin 2 hours before ovariectomy could improve the quality after transplantation of cryopreserved mouse OT., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Objective: To evaluate the feasibility of single-port laparoscopic myomectomy (SPLM) with intracoporeal suture-tying and transumbilical morcellation., Study Design: We retrospectively reviewed the medical records of 100 consecutive women who underwent SPLM without any combined surgery between January 2010 and July 2011. The medical records were reviewed and the clinical outcomes were analyzed. Single port entry was established using a wound retractor and a surgical glove. After myoma was enucleated from myometrium, intracorporeal suturing and knot tying was done for myometrium repair. Enucleated myoma was extracted through umbilicus using tissue morcellator. Factors investigated included operation time, blood loss, postoperative hospital stay and complications., Results: Mean patient age and BMI were 40.4±7.1 years and 23.1±3.5kg/m(2). Mean diameter of largest myoma and number of myomas were 6.6±2.2 (range 2-12) cm and 1.6±1.2 (range 1-7), respectively. Mean weight of myoma was 136.9±118.4 (range 4-684)g. Type of the main myomas were intramural (80/100), submucosal (5/100), subserosal (12/100), and intraligamentary (3/100). Location of the main myomas were anterior (36/100), posterior (24/100), fundal (30/100), and lateral (10/100). Mean operation time and estimated blood loss were 116.3±36.8min and 94.6±74.5mL. Mean hospital stay after the operation was 2±0.4 days. Conversion to double port surgery (one more trocar inserted on suprapubic site) was occurred in 1 patient due to severe pelvic adhesion. No patient experienced major complications, including bowel, ureter, or bladder injuries, or incisional hernia., Conclusion: SPLM is safe and acceptable for various myoma sites and sizes up to 12cm. Intracorporeal suture-tying and transumbilical morcellation are key technical points of SPLM., (Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.)

Study Objective: The objective of this study was to evaluate laparoendoscopic single-site myomectomy (LESS-M) for the surgical treatment of fibroids and to compare surgical outcomes and postoperative pain with conventional laparoscopic myomectomy (CLM)., Design: Retrospective study., Setting: University-based hospital., Patients: Data were obtained from medical records of patients who underwent LESS-M between August 2011 and June 2012. Considering the surgeon's learning curve for LESS-M, we collected the data after 100 LESS-M procedures were performed. The cases were compared with a historic cohort of patients who underwent CLM performed by the same surgeon between July 2008 and May 2009. A single experienced surgeon performed both procedures in all patients. A total of 118 patients who underwent LESS-M or CLM were included in the study (59 in the LESS-M group and 59 in the CLM group)., Interventions: None., Measurements and Main Results: We analyzed and compared patient basal characteristics and surgical outcomes between the 2 groups. There were no statistically significant differences in basal characteristics (i.e., age, body mass index, number and size of myomas, and type of the largest myoma) between the 2 groups. The surgical outcomes (i.e., operative time, estimated blood loss, postoperative hemoglobin drop, postoperative hospital stay, and postoperative pain scores) were not different statistically between the 2 groups. Moreover, patients did not experience major intraoperative complications. Postoperative complications were wound infections that occurred in 3 patients (2 in the LESS-M and 1 in the CLM groups)., Conclusion: LESS-M is feasible for less than 5 myomas and offers comparable surgical outcomes with those of CLM after the surgeon's initial learning curve., (Copyright © 2014 AAGL. Published by Elsevier Inc. All rights reserved.)

Objective: To investigate the effects of necrostatin-1 (Nec-1) supplementation on vitrification, warming and transplantation of ovarian tissue., Study Design: Ovaries from 4-week-old ICR mice were vitrified using a two-step procedure; ovaries were suspended in equilibration solution for 10min, and then mixed with vitrification solution for 5min. Ovaries were divided at random into three groups and 0 (control), 25 or 100μM Nec-1 was added to the vitrification solution. After warming, follicular morphology and apoptosis were assessed. For each group, a sample of vitrified, warmed ovaries was autotransplanted. The same dose of Nec-1 that was added to the vitrification solution was added to each warming solution and injected intraperitoneally. Follicular morphology and apoptosis of transplanted ovaries were assessed after 2 weeks., Results: After vitrification and warming, morphological analysis revealed that the intact follicle ratio was significantly higher in the Nec-1-treated groups compared with the control group (control, 45.1%; 25μM Nec-1, 51.7%; 100μM Nec-1, 57.9%). The rate of apoptosis was lower in the Nec-1 treated groups compared with the control group (control, 11.2%; 25μM Nec-1, 8.5%; 100μM Nec-1, 7.2%). After transplantation of the vitrified, warmed ovaries, morphological analysis revealed that the intact follicle ratio was significantly higher in the Nec-1 treated groups compared with the control group (control, 43.1%; 25μM Nec-1, 60.6%; 100μM Nec-1, 70.7%). The rate of apoptosis was lower in the Nec-1 treated groups compared with the control group (control, 5.3%; 25μM Nec-1, 2.5%; 100μM Nec-1, 2.0%)., Conclusions: Nec-1 supplementation during vitrification, warming and transplantation has beneficial effects on the survival of ovarian tissue. These results can help to improve ovarian tissue vitrification and transplantation protocols for fertility preservation., (Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.)

Study Question: What is the optimal vitrification protocol according to the cryoprotective agent (CPA) for ovarian tissue (OT) cryopreservation?, Summary Answer: The two-step protocol with 7.5% ethylene glycol (EG) and 7.5% dimethyl sulfoxide (DMSO) for 10 min then 20% EG, 20% DMSO and 0.5 M sucrose for 5 min showed the best results in mouse OT vitrification., What Is Known Already: Establishing the optimal cryopreservation protocol is one of the most important steps to improve OT survival. However, only a few studies have compared vitrification protocols with different CPAs and investigated the effect of in vitro culture (IVC) on vitrified-warmed OT survival. Some recent papers proposed that a combination of CPAs has less toxicity than one type of CPA. However, the efficacy of different types and concentrations of CPA are not yet well documented., Study Design, Size, Duration: A total of 644 ovaries were collected from 4-week-old BDF1 mice, of which 571 ovaries were randomly assigned to 8 groups and vitrified using different protocols according to CPA composition and the remaining 73 ovaries were used as controls. After warming, each of the eight groups of ovaries was further randomly divided into four subgroups and in vitro cultured for 0, 0.5, 2 and 4 h, respectively. Ovaries of the best two groups among the eight groups were autotransplanted after IVC., Participants/materials, Setting, Methods: The CPA solutions for the eight groups were composed of EDS, ES, ED, EPS, EF, EFS, E and EP, respectively (E, EG; D, DMSO; P, propanediol; S, sucrose; F, Ficoll). The IVC medium was composed of α-minimal essential medium, 10% fetal bovine serum and 10 mIU/ml follicle-stimulating hormone (FSH). Autotransplantation of vitrified-warmed OTs after IVC (0 to 4 h) using the EDS or ES protocol was performed, and the grafts were recovered after 3 weeks. Ovarian follicles were assessed for morphology, apoptosis, proliferation and FSH level., Main Results and the Role of Chance: The percentages of the morphologically intact (G1) and apoptotic follicles in each group at 0, 0.5, 2 and 4 h of IVC were compared. For G1 follicles at 0 and 4 h of IVC, the EDS group showed the best results at 63.8 and 46.6%, respectively, whereas the EP group showed the worst results at 42.2 and 12.8%, respectively. The apoptotic follicle ratio was lowest in the EDS group at 0 h (8.1%) and 0.5 h (12.7%) of IVC. All of the eight groups showed significant decreases in G1 follicles and increases in apoptotic follicles as IVC duration progressed. After autotransplantation, the EDS 0 h group showed a significantly higher G1 percentage (84.9%) than did the other groups (42.4-58.8%), while only the ES 4 h group showed a significant decrease in the number of proliferative cells (80.6%, 87.6-92.9%). However, no significant differences in apoptotic rates and FSH levels were observed between the groups after autotransplantation., Limitations, Reasons for Caution: The limitation of this study was the absence of in vitro fertilization using oocytes obtained from OT grafts, which should be performed to confirm the outcomes of ovarian cryopreservation and transplantation., Wider Implications of the Findings: We compared eight vitrification protocols according to CPA composition and found the EDS protocol to be the optimal method among them. The data presented herein will help improve OT cryopreservation protocols for humans or other animals.

Objective: To investigate the association of individual follicular fluid (FF) leptin and adiponectin levels with the quality of the corresponding oocyte and embryo., Methods: We prospectively enrolled 67 women who underwent controlled ovarian hyperstimulation with 89 FF samples. FF and the corresponding oocyte was obtained from a single dominant preovulatory follicle at the time of oocyte retrieval. Concentrations of leptin and adiponectin were measured by enzyme-linked immunosorbent assay in an individual follicle. The oocyte quality, fertilization rate, and corresponding embryo development were assessed., Results: The FF level of leptin was significantly associated with body mass index (r=0.334, p<0.01). The FF adiponectin level was significantly higher in the normal fertilization group than the abnormal fertilization group (p=0.009) in the non-obese women. A lower FF leptin level was associated with a trend toward mature oocytes, normal fertilization, and good embryo quality, although these relationships were not statistically significant. The leptin:adiponectin ratio of FF did not differ significantly according to oocyte and embryo quality. The quality of the oocyte and embryo was not associated with the FF leptin level tertile. However, the normal fertilization rate was positively associated with FF adiponectin level tertile. There was a trend towards improved oocytes and normal fertilization rates with the lowest tertile of the FF leptin:adiponectin ratio, but this difference was not statistically significant., Conclusion: Our results suggest that a high FF adiponectin concentration could be a predictor of normal fertilization. However, the FF leptin concentration and leptin:adiponectin ratio is not significantly related to oocyte maturity and corresponding embryo development.

Study Objective: To evaluate the safety and feasibility of single-port laparoscopic adnexal surgery during pregnancy., Design: A retrospective study (Canadian Task Force Classification III)., Setting: A university hospital., Patients: A total of 14 women with intrauterine pregnancy who underwent single-port laparoscopic surgery (SPLS)., Intervention: We retrospectively reviewed the medical records of 14 women with intrauterine pregnancy who underwent SPLS between November 2009 and September 2012 for the treatment of an adnexal mass., Measurements and Main Results: Factors investigated included the duration of surgery, differences between preoperative and postoperative hemoglobin, the size of the mass as measured by ultrasonography, major intraoperative or postoperative complications, conversion to multiport laparoscopy, and pregnancy outcomes. Single-port laparoscopic adnexal surgery was successfully performed in all 14 pregnant patients with an adnexal mass. The median gestational age at operation was 9 weeks (range, 4 weeks-17 weeks 4 days). Procedures included ovarian cystectomy with or without detorsion (n = 9), cyst aspiration with detorsion (n = 2), salpingectomy (n = 2), and salpingostomy (n = 1). The median duration of surgery was 59 minutes (range, 36-84 minutes). No major intraoperative or postoperative complications were encountered, and there was no need for conversion to multiport laparoscopic surgery. Five women had spontaneous vaginal delivery, and 5 women underwent cesarean delivery at term. One woman had preterm delivery at a gestational age of 24 weeks 5 days. In 1 case, abortion occurred 2 weeks after the operation., Conclusion: Based on these initial findings, SPLS appears to be a safe and technically feasible treatment for an adnexal mass during pregnancy., (Copyright © 2013 AAGL. Published by Elsevier Inc. All rights reserved.)

Objective: To investigate the effect of antifreeze protein (AFP) supplementation during mouse oocyte vitrification on the survival, fertilization and embryonic development., Design: Animal study., Setting: University laboratory., Animal(s): BDF-1 mice., Intervention(s): In vivo-matured metaphase II oocytes were vitrified with the use of CryoTop by two-step exposure to equilibrium and vitrification solution supplemented or not with 500 ng/mL AFP III., Main Outcome Measure(s): Postwarming survival, fertilization, embryonic development up to blastocyst in vitro, morphology of spindle and chromosome, membrane integrity, adenosine triphosphate (ATP) contents, and several gene expressions., Result(s): In the AFP-treated group, blastocyst formation rate was significantly higher and blastomere count with positive caspase was significantly lower compared with the nontreated group. Rate of intact spindle/chromosome, stable membrane, and ATP contents were significantly higher in AFP group. AFP group showed higher Mad2 and lower Eg5 gene expression. Both vitrification groups showed increased Hsf1, Zar1, and Zp1/Zp2 expression and decreased Hook1 and Zp3 expression compared with fresh control samples., Conclusion(s): Supplementation of AFP in vitrification medium has a protective effect on mouse oocytes for chilling injury; it can preserve spindle/membrane integrity and intracellular ATP contents. More stable spindle integrity in the AFP group may be associated with higher Mad2 and lower Eg5 gene expression., (Copyright © 2011 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.)

Objective: To investigates the effect of sphingosine-1-phosphate (S1P) supplementation on follicular integrity and apoptosis in vitrified-warmed mouse ovarian grafts., Study Design: Ovaries from 4-week-aged ICR mice were vitrified using a vitrification solution with or without 2 μM S1P. After warming, follicular normality was assessed by histological analysis and TUNEL assay. A part of ovaries vitrified with or without 2 μM S1P was transplanted, and 2 weeks later, gross and microscopic follicular morphology was assessed., Results: During vitrification and warming, inclusion of 2 μM S1P into the vitrification solution significantly raised the rate of morphologically intact follicles compared to controls (36.6% vs. 30.8%, p=0.047). This protective effect was profound especially in primordial follicles (45.5% vs. 34.6%, p=0.034). After transplantation of vitrified-warmed ovaries, the morphological integrity of primordial follicles was superior in the S1P-treated group (55.0% vs. 39.4%, p=0.035). The rates of non-apoptotic follicles (TUNEL-negative) were similar in the two groups in either non-transplanted or transplanted ovaries., Conclusion: Inclusion of S1P in the vitrification solution during transplantation of vitrified-warmed ovary had a beneficial effect on preservation of the primordial follicular pool., (Copyright © 2010 Elsevier Ireland Ltd. All rights reserved.)

Objective: To evaluate the ovarian reserve changes after laparoscopic cystectomy, we prospectively evaluated pre- and postoperative serum anti-Müllerian hormone (AMH) level, and ovarian volumes., Design: Prospective longitudinal study., Setting: University Hospital., Patient(s): Twenty women with benign ovarian masses participated; endometrioma [13], mature teratoma [6], and mucinous cystadenoma [1]. Seven patients had bilateral ovarian masses., Intervention(s): All patients had undergone laparoscopic ovarian cystectomy. Serum AMH levels were serially measured: preoperative, 1 week, 1 month, and 3 months after operation. Volumes of total ovary and ovarian mass were measured by 3D ultrasonography before operation., Main Outcome Measure(s): Postoperative serum AMH level and ovarian volume., Result(s): Median AMH level was 2.23 ng/mL (95% confidence interval [CI] 1.35-3.41 ng/mL) before operation, but reduced to 0.67 ng/mL (95% CI 0.44-1.70 ng/mL) at the first week postoperatively and then increased to 1.14 ng/mL (95% CI 0.79-2.36 ng/mL) in the first month and 1.50 ng/mL (95% CI 0.58-3.26 ng/mL) in the third month. The serum AMH level after 3 months postoperatively was recovered to about 65% of the preoperative level. The serum AMH level at postoperative 1 week was more decreased in endometrioma compared with nonendometrioma (33.9% vs. 69.2% of preoperative level), and in bilateral group compared with unilateral group (16.9% vs. 62.9%)., Conclusion(s): This study suggests that ovarian reserve could be reduced after laparoscopic cystectomy; however, it could be restored thereafter up to 3 months postoperative in reproductive women., (Copyright (c) 2010 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.)

Objective: To determine the sex ratio and risk of monozygotic twinning (MZT) after blastocyst transfer compared with cleavage-stage embryo transfer (ET) in fresh IVF cycles., Design: Systematic review and meta-analysis., Setting: University hospital center for reproductive medicine and IVF., Patient(s): Results of all reports from January 1995 to November 2007 with women undergoing nondonor fresh IVF. The sex ratio of 2,587 offspring and MZT rate in 40,917 cycles were included., Intervention(s): Cleavage or blastocyst stage for embryo transfer., Main Outcome Measure(s): Sex ratio and rate of MZT., Result(s): A meta-analysis was performed using four studies for sex ratio and nine studies for MZT. The results of the meta-analysis using a fixed effect model demonstrated a higher male-female ratio after blastocyst transfer compared with cleavage-stage ET (odds ratio [OR] 1.29, 95% confidence interval [CI] 1.10-1.51), and this difference was statistically significant. The risk of MZT after blastocyst transfer was significantly higher compared with cleavage-stage ET (OR 3.04, 95% CI 1.54-6.01)., Conclusion(s): The combined data presented in this meta-analysis suggest that blastocyst transfer appears to be associated with a sex ratio skewed in favor of males and an increased risk of MZT. The clinicians should provide this information to their infertility patients awaiting blastocyst transfer.

This study was performed to analyze retrospectively outcomes of stimulated in vitro fertilization (IVF) cycles where the gonadotropin-releasing hormone (GnRH) antagonist was omitted on ovulation triggering day. A total of 92 consecutive IVF cycles were included in 65 women who are undergoing ovarian stimulation with recombinant FSH. A GnRH antagonist, cetrorelix 0.25 mg/day, was started when leading follicle reached 14 mm in diameter until the day of hCG administration (Group A, 66 cycles) or until the day before hCG administration (Group B, 26 cycles). The duration of ovarian stimulation, total dose of gonadotropins, serum estradiol levels on hCG administration day, and the number of oocytes retrieved were not significantly different between the two groups. The total dose of GnRH antagonist was significantly lower in Group B compared to Group A (2.7+/-0.8 vs. 3.2+/-0.9 ampoules). There was no premature luteinization in the subjects. The proportion of mature oocytes (71.4% vs. 61.7%) and fertilization rate of mature (86.3+/-19.7% vs. 71.8+/-31.7%) was significantly higher in Group B. There were no significant differences in embryo quality and clinical pregnancy rates. Our results suggest that cessation of the GnRH antagonist on the day of hCG administration during a flexible multiple-dose protocol could reduce the total dose of GnRH antagonist without compromising IVF results.

Objective: To compare follicular fluid (FF) anti-Müllerian hormone (AMH) and inhibin B concentrations for GnRH agonist (GnRH-a) and GnRH antagonist cycles and to determine the correlations between FF AMH or inhibin B concentrations and controlled ovarian hyperstimulation (COH) outcomes., Design: Prospective comparative study., Setting: University hospital., Patient(s): Eighty-seven women who underwent COH cycles, either in the GnRH-a long-protocol group (n = 43) or the GnRH antagonist multiple-dose flexible-protocol group (n = 44)., Intervention(s): Follicular fluid was obtained from dominant follicles during oocyte retrieval, and FF AMH, inhibin B, E(2), and P concentrations were measured. Serum levels of AMH and inhibin B also were assessed on the day of oocyte retrieval., Main Outcome Measure(s): Follicular fluid AMH and inhibin B concentrations., Result(s): Concentrations of serum AMH and inhibin B and of FF AMH, inhibin B, E(2), and P were similar in the two groups. Follicular fluid AMH levels were found to be significantly correlated with age, gonadotropin dose, number of follicles on hCG day, and number of oocytes retrieved., Conclusion(s): Our results suggest that there is no significant difference in follicular microenvironment in terms of AMH and inhibin B secretion between GnRH-a and GnRH antagonist protocols and that FF AMH is a marker that reflects ovarian reserve and response to COH.