Oocyte maturity

卵母细胞成熟度
  • 文章类型: Journal Article
    目的评估体外受精-胚胎移植(IVF-ET)过程中,在正常卵巢储备患者中,周围触发女性生殖激素(FRHs)在预测卵母细胞成熟中的功效。
    使用医院数据库提取2020年1月至2021年9月的IVF-ET病例数据。女性生殖荷尔蒙的水平,包括雌二醇(E2),黄体生成素(LH),孕酮(P),和卵泡刺激素(FSH),最初是在基线时评估的,触发的那天,触发后的第二天,和取卵日。E2、LH、P,时间点1(触发日期和基线)和时间点2(触发日期之后和触发日期之后)之间的FSH分别定义为E2_RoV1/2、LH_RoV1/2、P_RoV1/2和FSH_RoV1/2。进行单变量和多变量回归来筛选周围触发FRHs以预测卵母细胞成熟。
    共有118名患者参加了我们的研究。单变量分析显示E2_RoV1与GnRH激动剂组的MII卵母细胞比率之间存在显著关联(p<0.05),但在GnRH拮抗剂方案组中没有。相反,P_RoV2作为两个方案组中MII卵母细胞率的潜在预测因子(p<0.05)。多变量分析证实P_RoV2在预测两组卵母细胞成熟率中的意义(p<0.05)。而E2_RoV1在两组中的相关性均不显著。然而,在GnRH激动剂方案组中的高P_RoV2亚组内,没有观察到相关性是显著的。GnRH激动剂方案组的C指数为0.83(95%CI[0.73-0.92]),GnRH拮抗剂方案组为0.77(95%CI[0.63-0.90])。ROC曲线分析进一步支持了模型的令人满意的性能,GnRH激动剂方案组的曲线下面积(AUC)值为0.79,GnRH拮抗剂方案组为0.81。
    P_RoV2对GnRH激动剂和GnRH拮抗剂方案组的卵母细胞成熟均显示出显著的预测价值,这增强了对评估卵母细胞成熟的理解,并为正常卵巢储备患者在IVF-ET期间控制性超促排卵的个体化治疗方案提供了信息。
    UNASSIGNED: To evaluate the efficacy of peri-trigger female reproductive hormones (FRHs) in the prediction of oocyte maturation in normal ovarian reserve patients during the in vitro fertilization-embryo transfer (IVF-ET) procedure.
    UNASSIGNED: A hospital database was used to extract data on IVF-ET cases from January 2020 to September 2021. The levels of female reproductive hormones, including estradiol (E2), luteinizing hormone (LH), progesterone (P), and follicle-stimulating hormone (FSH), were initially evaluated at baseline, the day of the trigger, the day after the trigger, and the day of oocyte retrieval. The relative change in E2, LH, P, FSH between time point 1 (the day of trigger and baseline) and time point 2 (the day after the trigger and day on the trigger) was defined as E2_RoV1/2, LH_RoV1/2, P_RoV1/2, and FSH_RoV1/2, respectively. Univariable and multivariable regression were performed to screen the peri-trigger FRHs for the prediction of oocyte maturation.
    UNASSIGNED: A total of 118 patients were enrolled in our study. Univariable analysis revealed significant associations between E2_RoV1 and the rate of MII oocytes in the GnRH-agonist protocol group (p < 0.05), but not in the GnRH-antagonist protocol group. Conversely, P_RoV2 emerged as a potential predictor for the rate of MII oocytes in both protocol groups (p < 0.05). Multivariable analysis confirmed the significance of P_RoV2 in predicting oocyte maturation rate in both groups (p < 0.05), while the association of E2_RoV1 was not significant in either group. However, within the subgroup of high P_RoV2 in the GnRH-agonist protocol group, association was not observed to be significant. The C-index was 0.83 (95% CI [0.73-0.92]) for the GnRH-agonist protocol group and 0.77 (95% CI [0.63-0.90]) for the GnRH-antagonist protocol group. The ROC curve analysis further supported the satisfactory performance of the models, with area under the curve (AUC) values of 0.79 for the GnRH-agonist protocol group and 0.81 for the GnRH-antagonist protocol group.
    UNASSIGNED: P_RoV2 showed significant predictive value for oocyte maturation in both GnRH-agonist and GnRH-antagonist protocol groups, which enhances the understanding of evaluating oocyte maturation and inform individualized treatment protocols in controlled ovarian hyperstimulation during IVF-ET for normal ovarian reserve patients.
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  • 文章类型: Journal Article
    卵巢组织玻璃化被广泛用于青春期前和青春期女性癌症患者的生育力保存。目前的文献包括自体移植后成功怀孕和活产的报道。然而,玻璃化过程对卵巢组织中对卵母细胞成熟和早期胚胎发育至关重要的卵丘-壁颗粒细胞(C-mGCs)-体细胞的影响尚不清楚。本研究通过定量检测生长分化因子9(GDF-9)的表达,探讨玻璃化对C-mGCs细胞功能的影响。骨形态发生蛋白15(BMP-15),卵泡刺激素受体(FSHR),黄体生成素受体(LHR),连接蛋白37幸存者,和胱天蛋白酶3。
    从参与体外受精计划的38名多囊卵巢综合征妇女中获得成熟和未成熟的C-mGC。然后将C-mGC分成两组:新鲜的和玻璃化的。使用实时定量聚合酶链反应评估靶基因的表达水平。
    玻璃化后,GDF-9表达在成熟和未成熟的C-mGCs中显著降低,随着0.2倍和0.1倍的变化,分别(p<0.01)。同样,FSHR在成熟和未成熟组中的表达减少了0.1倍和0.02倍,分别,玻璃化后(p<0.01)。其他基因的表达水平,包括BMP-15,LHR,连接蛋白37幸存者,和半胱天冬酶3,在各实验组中保持相似(p>0.05)。
    玻璃化可能通过加热后C-mGCs中GDF-9和FSHR表达减少而损害卵母细胞成熟。
    UNASSIGNED: Ovarian tissue vitrification is widely utilized for fertility preservation in prepubertal and adolescent female patients with cancer. The current literature includes reports of successful pregnancy and live birth following autografting. However, the effects of the vitrification process on cumulus-mural granulosa cells (C-mGCs)-somatic cells in ovarian tissue crucial for oocyte maturation and early embryonic development-remain unclear. This study was conducted to explore the impact of vitrification on the cellular function of C-mGCs by quantifying the expression of growth differentiation factor 9 (GDF-9), bone morphogenetic protein 15 (BMP-15), follicle-stimulating hormone receptor (FSHR), luteinizing hormone receptor (LHR), connexin 37, survivin, and caspase 3.
    UNASSIGNED: Mature and immature C-mGCs were obtained from 38 women with polycystic ovary syndrome who participated in an in vitro fertilization program. The C-mGCs were then divided into two groups: fresh and vitrified. The expression levels of target genes were assessed using real-time quantitative polymerase chain reaction.
    UNASSIGNED: After vitrification, GDF-9 expression was significantly decreased among both mature and immature C-mGCs, with 0.2- and 0.1-fold changes, respectively (p<0.01). Similarly, FSHR expression in the mature and immature groups was reduced by 0.1- and 0.02-fold, respectively, following vitrification (p<0.01). The expression levels of the other genes, including BMP-15, LHR, connexin 37, survivin, and caspase 3, remained similar across the examined groups (p>0.05).
    UNASSIGNED: Vitrification may compromise oocyte maturation through reduced GDF-9 and FSHR expression in C-mGCs after warming.
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  • 文章类型: Journal Article
    目的:促性腺激素释放激素激动剂(GnRHa)排卵后,卵母细胞恢复率(ORR)差和卵母细胞不成熟的潜在危险因素是什么?
    结论:促性腺激素释放激素激动剂(GnRHa)触发后卵巢储备功能降低和LH水平是ORR差的危险因素。较高的BMI和抗苗勒管激素(AMH)水平是卵母细胞成熟率(OMR)低下的危险因素。
    背景:使用GnRHa引发排卵的情况正在增加。然而,一些患者在GnRHa触发后可能出现不良反应.这种次优响应可以指任何负终点,例如卵母细胞恢复不理想,卵母细胞不成熟,或者空卵泡综合征.对于一些作者来说,对GnRHa触发的次优反应是指触发后LH和/或孕酮水平次优。几项研究调查了人口统计学的组合,临床,和内分泌特征在治疗过程的不同阶段,可能会影响GnRHa触发剂的疗效,因此涉及不良的内分泌反应或效率,但没有共识存在。
    方法:2015年至2021年的双中心回顾性队列研究(N=1747)。
    方法:纳入所有18-43岁接受控制性超促排卵和单独使用GnRHa(曲普瑞林0.2mg)进行ICSI或卵母细胞冷冻保存的排卵触发的患者。ORR定义为在触发当天回收的卵母细胞总数与>12mm的卵泡数的比率。OMR定义为成熟卵母细胞的数量与回收的卵母细胞的数量的比率。采用后向选择方法的logistic回归模型进行危险因素分析。赔率比(OR)以其双侧95%置信区间显示。
    结果:在多变量分析中,触发后12小时的初始窦卵泡计数和LH水平与ORR不良(即低于第10百分位数)呈负相关(OR:0.61[95%CI:0.42-0.88];P=0.008和OR:0.86[95%CI:0.76-0.97];P=0.02,分别).在触发后12小时的LH水平与不良ORR之间发现了非线性关系,但未发现LH阈值。总共25.3%的患者患有卵母细胞不成熟(即OMR<75%)。在多变量分析中,BMI和AMH水平与OMR<75%呈负相关(OR:4.34[95%CI:1.96-9.6];P<0.001和OR:1.22[95%CI:1.03-1.12];P=0.015,分别)。与无预处理相比,抗促性腺激素预处理降低了OMR<75%的风险(OR:0.72[95%CI:0.57-0.91];P=0.02)。
    结论:我们的研究受到回顾性设计和排除hCG再触发者的限制。然而,这只发生在六个周期中。我们也无法收集有关预处理持续时间和冲洗期持续时间的信息。
    结论:在临床实践中,为了避免糟糕的ORR,在BMI较高和/或卵巢储备较低的患者中,不应仅考虑GnRHa触发。与卵巢过度刺激综合征的风险平衡。在低的12小时后触发LH水平的情况下,执业医师必须意识到ORR不良的风险,可以考虑重新触发hCG。
    背景:无。
    背景:不适用。
    OBJECTIVE: What are the potential risk factors for poor oocyte recuperation rate (ORR) and oocyte immaturity after GnRH agonist (GnRHa) ovulation triggering?
    CONCLUSIONS: Lower ovarian reserve and LH levels after GnRHa triggering are risk factors of poor ORR. Higher BMI and anti-Müllerian hormone (AMH) levels are risk factors of poor oocyte maturation rate (OMR).
    BACKGROUND: The use of GnRHa to trigger ovulation is increasing. However, some patients may have a suboptimal response after GnRHa triggering. This suboptimal response can refer to any negative endpoint, such as suboptimal oocyte recovery, oocyte immaturity, or empty follicle syndrome. For some authors, a suboptimal response to GnRHa triggering refers to a suboptimal LH and/or progesterone level following triggering. Several studies have investigated a combination of demographic, clinical, and endocrine characteristics at different stages of the treatment process that may affect the efficacy of the GnRHa trigger and thus be involved in a poor endocrine response or efficiency but no consensus exists.
    METHODS: Bicentric retrospective cohort study between 2015 and 2021 (N = 1747).
    METHODS: All patients aged 18-43 years who underwent controlled ovarian hyperstimulation and ovulation triggering by GnRHa alone (triptorelin 0.2 mg) for ICSI or oocyte cryopreservation were included. The ORR was defined as the ratio of the total number of retrieved oocytes to the number of follicles >12 mm on the day of triggering. The OMR was defined as the ratio of the number of mature oocytes to the number of retrieved oocytes. A logistic regression model with a backward selection method was used for the analysis of risk factors. Odds ratios (OR) are displayed with their two-sided 95% confidence interval.
    RESULTS: In the multivariate analysis, initial antral follicular count and LH level 12-h post-triggering were negatively associated with poor ORR (i.e. below the 10th percentile) (OR: 0.61 [95% CI: 0.42-0.88]; P = 0.008 and OR: 0.86 [95% CI: 0.76-0.97]; P = 0.02, respectively). A nonlinear relationship was found between LH level 12-h post-triggering and poor ORR, but no LH threshold was found. A total of 25.3% of patients suffered from oocyte immaturity (i.e. OMR < 75%). In the multivariate analysis, BMI and AMH levels were negatively associated with an OMR < 75% (OR: 4.34 [95% CI: 1.96-9.6]; P < 0.001 and OR: 1.22 [95% CI: 1.03-1.12]; P = 0.015, respectively). Antigonadotrophic pretreatment decreased the risk of OMR < 75% compared to no pretreatment (OR: 0.72 [95% CI: 0.57-0.91]; P = 0.02).
    CONCLUSIONS: Our study is limited by its retrospective design and by the exclusion of patients who had hCG retriggers. However, this occurred in only six cycles. We were also not able to collect information on the duration of pretreatment and the duration of wash out period.
    CONCLUSIONS: In clinical practice, to avoid poor ORR, GnRHa trigger alone should not be considered in patients with higher BMI and/or low ovarian reserve, balanced by the risk of ovarian hyperstimulation syndrome. In the case of a low 12-h post-triggering LH level, practicians must be aware of the risk of poor ORR, and hCG retriggering could be considered.
    BACKGROUND: None.
    BACKGROUND: N/A.
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  • 文章类型: Journal Article
    目的:确定触发当天的最小卵泡体积,该最小卵泡体积对应于通过个性化卵泡穿刺取卵时的成熟卵母细胞,并计算从排卵诱导到SonoAVC™卵泡取卵的平均卵泡生长。
    方法:对53名接受试管婴儿的妇女进行前瞻性观察研究,通过三维超声可以明确识别触发和取卵时的一个或多个卵泡。
    方法:大学附属的私人IVF中心。
    方法:最终样品包括来自14个卵母细胞供体和39名患者的206个卵泡。
    方法:在触发和取卵时使用SonoAVC™卵泡进行三维超声。相同的操作者选择在两次扫描中容易识别的卵泡,并验证它们易于单独抽吸。卵泡被单独刺穿,记录卵母细胞的实际抽吸量和成熟阶段。
    方法:主要结果是触发当天卵泡体积与卵母细胞成熟期之间的关系。次要结果是通过SonoAVC™卵泡测量的从触发当天到卵母细胞取出当天的卵泡生长速率。
    结果:在触发当天选择206个卵泡。其中,5在取卵当天无法识别,可能是由于卵泡破裂(平均体积4cm3,范围2-7),并且在48中未获得卵母细胞。研究了153个卵泡中卵泡体积与卵母细胞成熟度的关系,125(82%)成熟和28(18%)未成熟。ROC曲线显示0.73的曲线下面积(95CI:0.65-0.80,p<0.001)。>0.56cm3的卵泡体积是具有最高Youden指数的切点,其具有85%的灵敏度和64%的特异性以预测卵母细胞成熟度。在53%的病例中,从触发到取卵的平均卵泡生长为26-50%。
    结论:触发时>0.56cm3的卵泡体积是卵母细胞成熟度敏感性和特异性之间最佳平衡的临界点。>2-3cm3的卵泡在取卵前可能会自发破裂。鉴于这些发现,我们提出了新的基于体积的触发标准:70%的卵泡>0.6cm3,优势卵泡在2~3cm3之间.这些需要随机对照试验的验证。
    OBJECTIVE: To determine the minimum follicular volume on the day of trigger that will correspond to a mature oocyte at egg retrieval by individualized follicular puncture and to calculate the mean follicular growth from ovulation induction to egg retrieval using SonoAVCfollicle.
    METHODS: A prospective observational study of 53 women undergoing in vitro fertilization, in which it was possible to identify unequivocally one or more follicles at trigger and egg retrieval using three-dimensional ultrasound.
    METHODS: University-affiliated private in vitro fertilization center.
    METHODS: The final sample included 206 follicles from 14 oocyte donors and 39 patients.
    METHODS: A three-dimensional ultrasound with SonoAVCfollicle was performed at trigger and egg retrieval. The same operator selected follicles that were identified easily on both scans and verified that they were apt to be aspirated individually. Follicles were punctured individually, recording the real aspirated volume and the maturity stage of the oocyte.
    METHODS: The primary outcome was the relationship between follicular volume on the day of the trigger and the oocyte maturity stage. The secondary outcome was the rate of follicular growth from the day of trigger to the day of oocyte retrieval, as measured using SonoAVCfollicle.
    RESULTS: On the day of trigger 206, follicles were selected. Of these, 5 could not be identified on the day of oocyte retrieval, probably because of follicular rupture (mean volume: 4 cm3, range: 2-7 cm3), and in 48, an oocyte was not obtained. The relationship between follicular volume and oocyte maturity was studied in 153 follicles: 125 (82%) contained mature and 28 (18%) contained immature oocytes. Receiver operating characteristic curves showed an area under the curve value of 0.73 (95% confidence interval: 0.65-0.80). A follicular volume of >0.56 cm3 is the cutoff point, with the highest Youden index having a sensitivity of 85% and a specificity of 64% to predict oocyte maturity. The mean follicular growth from trigger to egg retrieval was 26%-50% in 53% of cases.
    CONCLUSIONS: A follicular volume of >0.56 cm3 at trigger is the cutoff point with the optimal balance between sensitivity and specificity for oocyte maturity. Follicles of >2-3 cm3 may undergo spontaneous rupture before egg retrieval. Given these findings, we propose new volume-based criteria for trigger: 70% of follicles of >0.6 cm3 and dominant follicles between 2 and 3 cm3. These findings need validation by randomized controlled trials.
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  • 文章类型: Journal Article
    卵泡生成中的血管生成有助于自然和体外受精(IVF)周期中的卵母细胞发育能力。因此,卵泡发生过程中卵泡液(FF)中关键血管生成因子的鉴定具有临床意义,对体外受精具有重要意义。这项研究旨在确定FF中关键的血管生成因子,以预测体外受精过程中的卵母细胞成熟度。
    招募了40名在首次体外受精治疗中使用GnRH拮抗剂方案进行卵巢刺激的参与者。每个病人,两个卵泡样本(一个排卵前卵泡,>18毫米;一个中窦卵泡,<14mm)在卵母细胞取出过程中没有潮红的情况下收集。总的来说,从40名患者收集了80份FF样品。通过Luminex高性能测定分析FF中血管生成相关蛋白的表达谱。记录的患者数据包括窦卵泡计数,抗苗勒管激素,年龄,BMI。在月经周期第2天,触发日收集血清样本,和取卵日。通过化学发光法测量激素浓度,包括第2天的FSH/LH/E2/P4,触发天的E2/LH/P4和取回天的E2/LH/P4。
    10种血管生成因子在FF中高表达:eotaxin,Gro-α,IL-8,IP-10,MCP-1,MIG,PAI-1(Serpin),VEGF-A,CXCL-6和HGF。eotaxin的浓度,IL-8、MCP1、PAI-1、VEGF-A在排卵前卵泡明显高于中腔卵泡,排卵前卵泡中Gro-α和CXCL-6的表达水平低于中腔卵泡(p<0.05)。Logistic回归和受试者工作特征(ROC)分析显示,VEGF-A,eotaxin,和CXCL-6是卵母细胞成熟度的三个最强预测因子。VEGF-A和CXCL-6的组合比其他组合以更高的灵敏度(91.7%)和特异性(72.7%)预测卵母细胞成熟度。
    我们的研究结果表明VEGF-A,eotaxin,FF中的CXCL-6浓度与从窦中期到排卵前期的卵母细胞成熟度密切相关。VEGF-A和CXCL-6的组合在体外受精过程中显示出相对较好的卵母细胞成熟度预测率。
    Angiogenesis in folliculogenesis contributes to oocyte developmental competence in natural and in vitro fertilization (IVF) cycles. Therefore, the identification of key angiogenic factors in follicular fluid (FF) during folliculogenesis is clinically significant and important for in vitro fertilization. This study aims to identify the key angiogenic factors in FF for predicting oocyte maturity during in vitro fertilization.
    Forty participants who received ovarian stimulation using a GnRH antagonist protocol in their first in vitro fertilization treatment were recruited. From each patient, two follicular samples (one preovulatory follicle, > 18 mm; one mid-antral follicle, < 14 mm) were collected without flushing during oocyte retrieval. In total, 80 FF samples were collected from 40 patients. The expression profiles of angiogenesis-related proteins in FF were analyzed via Luminex high-performance assays. Recorded patient data included antral follicle count, anti-müllerian hormone, age, and BMI. Serum samples were collected on menstrual cycle day 2, the trigger day, and the day of oocyte retrieval. Hormone concentrations including day 2 FSH/LH/E2/P4, trigger day E2/LH/P4, and retrieval day E2/LH/P4 were measured by chemiluminescence assay.
    Ten angiogenic factors were highly expressed in FF: eotaxin, Gro-α, IL-8, IP-10, MCP-1, MIG, PAI-1 (Serpin), VEGF-A, CXCL-6, and HGF. The concentrations of eotaxin, IL-8, MCP1, PAI-1, and VEGF-A were significantly higher in preovulatory follicles than those in mid-antral follicles, while the Gro-α and CXCL-6 expressional levels were lower in preovulatory than in mid-antral follicles (p < 0.05). Logistic regression and receiver operating characteristic (ROC) analysis revealed that VEGF-A, eotaxin, and CXCL-6 were the three strongest predictors of oocyte maturity. The combination of VEGF-A and CXCL-6 predicted oocyte maturity with a higher sensitivity (91.7%) and specificity (72.7%) than other combinations.
    Our findings suggest that VEGF-A, eotaxin, and CXCL-6 concentrations in FF strongly correlate with oocyte maturity from the mid-antral to preovulatory stage. The combination of VEGF-A and CXCL-6 exhibits a relatively good prediction rate of oocyte maturity during in vitro fertilization.
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  • 文章类型: Journal Article
    To identify relationships between the size of punctured ovarian follicles and subsequent embryology outcomes.
    Prospective observational cohort study.
    Private fertility center.
    One hundred fifty-seven oocyte retrievals performed during the study period.
    The diameter of punctured follicles was ultrasonically measured during routine oocyte collection. The resulting embryos were group-cultured to the blastocyst stage and classified into 8 groups according to follicle size (≤9.5, 10-12.5, 13-15.5, 16-18.5, 19-21.5, 22-24.5, 25-27.5, and ≥28 mm).
    Rate of good-quality blastocysts per follicle puncture.
    This study included 4,539 follicle punctures, 2,348 oocytes, 1,772 mature oocytes, 1,258 bipronuclear (2pn) oocytes, and 571 good-quality blastocysts derived from 157 oocyte retrievals. The per-puncture yields of oocytes, mature oocytes, 2pn oocytes, and good-quality blastocysts were associated with the size of the punctured follicle. The rates of good-quality blastocysts per punctured follicle were 2.2% (≤9.5 mm), 6.2% (10-12.5 mm), 11.9% (13-15.5 mm), 14.5% (16-18.5 mm), 18.9% (19-21.5 mm), 17.5% (22-24.5 mm), 15.9% (25-27.5 mm), and 16.0% (≥28 mm). When compared with the overall average, punctures of follicles in groups ≤12.5 mm in diameter had significantly inferior yields of good-quality blastocysts, whereas punctures of follicles in groups 19-24.5 mm in diameter were associated with significantly greater than average yields of good-quality blastocysts. Other groups did not differ significantly from average. No correlation was observed between follicle diameter and ploidy of biopsied blastocysts.
    Punctures of follicles ≤12.5 mm in diameter rarely result in good-quality blastocysts. The yield of good-quality blastocysts progressively increases with follicle size up to approximately 19 mm in diameter, with no substantial decline above that size. The ploidy of the blastocysts that form appears to be unaffected by follicle size.
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  • 文章类型: Journal Article
    我们基于不同的高分辨率质谱仪和LC设置,提出了两个独立的无标签定量工作流程。在所使用的仪器之后称为:四轨道阱(nano-LC)和三四TOF(micro-LC)及其对人类卵泡液蛋白质组分析的定向适应。我们使用各种样品制备方法在每个不同的工作流程中鉴定了约1000种蛋白质。在总蛋白质方法的帮助下,我们能够获得每个工作流程的绝对蛋白质浓度.在一项对来自四个供体的与不同卵母细胞质量状态相关的20个样本的初步研究中,455和215蛋白质通过四轨道和三重四TOF工作流程进行了定量。分别。从两个工作流程获得的浓度值在显著程度上相关。我们发现测试组之间蛋白质折叠变化的两个工作流程的合理一致性,产生了与卵母细胞成熟和胚泡发育相关的20和22种蛋白质的统一列表,分别。Quad-Orbitrap工作流程最适合进行深入分析,而无需进行大量分馏,尤其是低丰度蛋白质组,而TripleQuad-TOF工作流程允许一种更稳健的方法,在构建全面的光谱库的最初努力之后,随着分析样本数量的增加,该方法具有更大的提高有效性的潜力。
    We present two separate label-free quantitative workflows based on different high-resolution mass spectrometers and LC setups, which are termed after the utilized instrument: Quad-Orbitrap (nano-LC) and Triple Quad-TOF (micro-LC) and their directed adaptation toward the analysis of human follicular fluid proteome. We identified about 1000 proteins in each distinct workflow using various sample preparation methods. With assistance of the Total Protein Approach, we were able to obtain absolute protein concentrations for each workflow. In a pilot study of twenty samples linked to diverse oocyte quality status from four donors, 455 and 215 proteins were quantified by the Quad-Orbitrap and Triple Quad-TOF workflows, respectively. The concentration values obtained from both workflows correlated to a significant degree. We found reasonable agreement of both workflows in protein fold changes between tested groups, resulting in unified lists of 20 and 22 proteins linked to oocyte maturity and blastocyst development, respectively. The Quad-Orbitrap workflow was best suited for an in-depth analysis without the need of extensive fractionation, especially of low abundant proteome, whereas the Triple Quad-TOF workflow allowed a more robust approach with a greater potential to increase in effectiveness with the growing number of analyzed samples after the initial effort of building a comprehensive spectral library.
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  • 文章类型: Case Reports
    体外受精(IVF)周期取消的最常见原因是缺乏可用于胞浆内精子注射(ICSI)的优质配子。在这里,我们介绍了受阻塞性无精子症影响的夫妇的成功生育治疗以及对控制性卵巢刺激的次优反应。由于传统方法似乎无法有效克服双方的具体问题,有针对性的干预措施,即,(1)精子活力的药理增强和(2)偏振光显微镜(PLM)引导下ICSI时间的优化,用于挽救周期,仅回收未成熟的卵母细胞和不运动的睾丸精子。茶碱治疗有助于从冷冻保存的睾丸组织中选择可行的精子。当传统的刺激方案无法产生成熟的卵子时,采用非侵入性纺锤体成像将精子注射时间调整到体外挤出极体卵母细胞的成熟阶段。12个晚熟卵母细胞受精产生5个受精卵,都发育成胚泡。受精后第5天,将一个胚胎转移到子宫中,另外3个质量好的胚泡被玻璃化以备后用。怀孕导致足月分娩健康的孩子。该病例表明,应考虑超出标准IVF方案的个体化,以最大程度地提高预后不良患者用自己的配子实现妊娠的机会。
    The most common reason for in vitro fertilization (IVF) cycle cancelation is a lack of quality gametes available for intracytoplasmic sperm injection (ICSI). Here we present the successful fertility treatment of the couple affected by obstructive azoospermia combined with suboptimal response to controlled ovarian stimulation. Since the conventional approach appeared ineffective to overcome both partners\' specific problems, the targeted interventions, namely, (1) pharmacological enhancement of sperm motility and (2) polarized light microscopy (PLM)-guided optimization of ICSI time, were applied to rescue the cycle with only immature oocytes and immotile testicular sperm retrieved. The treatment with theophylline aided the selection of viable spermatozoa derived from cryopreserved testicular tissue. When the traditional stimulation protocol failed to produce mature eggs, non-invasive spindle imaging was employed to adjust the sperm injection time to the maturational stage of oocytes extruding a polar body in vitro. The fertilization of 12 late-maturing oocytes yielded 5 zygotes, which all developed into blastocysts. One embryo was transferred into the uterus on day 5 post-fertilization, and another 3 good quality blastocysts were vitrified for later use. The pregnancy resulted in a full-term delivery of a healthy child. This case demonstrates that the individualization beyond the standard IVF protocols should be considered to maximize the chance of poor-prognosis patients to achieve pregnancy with their own gametes.
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  • 文章类型: Journal Article
    UNASSIGNED: The aim is to study the effect of follicle-stimulating hormone (FSH) administration on the day of human chorionic gonadotropin (hCG) trigger on the assisted reproductive technique (ART) outcomes in in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) cycles.
    UNASSIGNED: Retrospective cohort study was conducted in the ART center of our hospital.
    UNASSIGNED: Two hundred and ninety IVF/ICSI cycles performed between September 2012 and August 2017 were included in the study. Patients who received 375 IU of FSH on the day of hCG trigger (149 cycles) were compared with those who did not receive FSH on the day of trigger (141 cycles).
    UNASSIGNED: Chi-square test and Student\'s t-test were used.
    UNASSIGNED: The FSH co-administered group had a significantly higher number of oocytes retrieved, mature oocytes, and fertilization rate compared to those who did not receive FSH on the day of trigger (p < 0.001). The total number of embryos, the number of grade 1 embryos and the number of embryos available for cryopreservation were also significantly higher in the FSH administered group (p < 0.001). Implantation rate, clinical pregnancy rate, and live birth rate were not significantly different between the two groups.
    UNASSIGNED: This study has shown that FSH administration on the day of the trigger may be considered in IVF cycles receiving hCG trigger to improve the oocyte recovery and maturity if the patient is not at increased risk of ovarian hyperstimulation and serum estradiol on the day of the trigger is <4500 pg/ml. However, there is only an increase in the total number of oocytes retrieved and the number of mature oocytes but no significant change in the implantation, clinical pregnancy, and live birth rates.
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  • 文章类型: Journal Article
    成熟卵母细胞的数量是辅助生殖技术(ART)成功的关键因素。在卵巢刺激期间施用外源性促性腺激素以使可用于受精的卵母细胞的数量最大化。在刺激期间,监测是评估个人反应的强制性要求,以避免治疗并发症,并协助确定最终卵母细胞成熟和取卵的最佳日期。刺激期间的常规监测包括经阴道超声检查和血清雌二醇(E2)的测量。由于不同大小的卵泡的多卵泡生长,血清E2水平通常是超生理的,通常是可变的,在卵巢刺激期间进行E2测量不可靠,作为卵母细胞成熟度的决定因素。与血清E2相反,一旦达到12-15毫米的最小卵泡大小,血清抑制素A水平就会增加。由于这一事实,血清抑制素A水平与超声监测相结合,可以提供更可靠的参数,以确定最终卵母细胞成熟的最佳卵泡大小,因为只有大小为12毫米及以上的卵泡会导致血清抑制素A水平。这种前瞻性的观察,横断面研究表明,在最终卵母细胞成熟当天,血清抑制素A与卵泡数≥15mm(0.72)以及回收和成熟卵母细胞数(分别为ρ0.82/0.77)密切相关,而血清E2与上述参数中度相关(分别为ρ0.64/0.69/0.69)。抑制素A的曲线下面积(AUC)为0.91,与E2的AUC为0.84相比,在阈值数量≥10个成熟卵母细胞的情况下,抑制素A可被认为是触发药物治疗最佳时机的更好预测指标.从该数据可以得出结论,与E2相比,血清抑制素A与经阴道超声监测相结合可能是触发时机决策过程中更强大的工具。
    The number of mature oocytes is a key factor in the success of Assisted Reproductive Techniques (ART). Exogenous gonadotropins are administered during ovarian stimulation in order to maximize the number of oocytes available for fertilization. During stimulation, monitoring is mandatory to evaluate individual response, to avoid treatment complications and assist in the determination of the optimal day for final oocyte maturation and oocyte retrieval. Routine monitoring during stimulation includes transvaginal ultrasound examinations and measurement of serum estradiol (E2). Due to multifollicular growth of follicles of varying size, serum E2 levels are commonly supraphysiological and often variable, rendering E2-measurement during ovarian stimulation unreliable as a determinant of oocyte maturity. In contrast to serum E2, serum Inhibin A levels increase once a minimum follicle size of 12-15 mm is achieved. Due to this fact, serum Inhibin A levels could present in combination with ultrasound monitoring a more reliable parameter to determine the optimal follicle size for final oocyte maturation, as only follicles with a size of 12 mm and beyond will contribute to the serum Inhibin A level. This prospective observational, cross-sectional study demonstrates, that on the day of final oocyte maturation serum Inhibin A is strongly correlated to the number of follicles ≥15 mm (0.72) and to the number of retrieved and mature oocytes (ρ 0.82/0.77, respectively), whereas serum E2 is moderately correlated to the parameters mentioned above (ρ 0.64/0.69/0.69, respectively). With an area under the curve (AUC) of 0.91 for Inhibin A, compared to an AUC of 0.84 for E2, Inhibin A can be regarded as a better predictor for the optimal timing of trigger medication with a threshold number of ≥10 mature oocytes. It can be concluded from this data that serum Inhibin A in combination with transvaginal ultrasound monitoring may be a more powerful tool in the decision making process on trigger timing as compared to E2.
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