卵巢储备功能低下患者血清AMH、Betatrophin水平与卵巢反应性、IVF-ET妊娠结局关系

目的:探讨卵巢储备功能低下(DOR)患者血清抗苗勒管激素(AMH)、促代谢因子(Betatrophin)水平与卵巢反应性及体外受精-胚胎移植(IVF-ET)妊娠结局的关系。方法:收集2019年12月-2022年12月在本院行IVF-ET的DOR患者137例临床资料,根据卵巢反应性将患者分为低反应组(n=112)、正常反应组(n=16)、高反应组(n=9);根据妊娠结局将患者分为妊娠成功组(n=44)和妊娠失败组(n=93)。酶联免疫吸附法检测血清AMH、Betatrophin水平,受试者工作特性(ROC)曲线评估2项指标预测IVF-ET妊娠结局价值;多因素logistic回归分析影响妊娠的因素。结果:低反应组、正常组、高反应组血清AMH水平(0.49±0.13 ng/ml、0.98±0.21 ng/ml、1.05±0.26 ng/ml)依次升高,血清Betatrophin水平(156.95±16.33 pg/ml、112.17±13.42 pg/ml、92.64±11.03 pg/ml)依次降低;妊娠成功组血清AMH水平(1.07±0.36 ng/ml)高于妊娠失败组(0.34±0.19 ng/ml),Betatrophin水平(136.29±14.42 pg/ml)低于妊娠失败组(216.16±21.05 pg/ml)(均P<0.05)。血清AMH、Betatrophin预测DOR患者IVF-ET妊娠结局的曲线下面积(AUC)(95%CI)别为0.857、0.771,两项指标联合预测的AUC为0.904。多因素logistic回归分析显示,卵泡刺激素/促黄体生成素比值≥2、卵巢低反应、AMH≤0.71 ng/ml、Betatrophin≥176.23 pg/ml是DOR患者IVF-ET妊娠失败的独立危险因素(P<0.05)。结论:血清AMH、Betatrophin水平与DOR患者卵巢反应性、IVF-ET妊娠结局有关,二者有望作为预测DOR患者IVF-ET妊娠结局的生物标记物。

Effects of LPA on the development of sheep in vitro fertilized embryos and attempt to establish sheep embryonic stem cells

Lysophosphatidic acid(LPA)is a small molecule glycerophospholipid,which regulates multiple downstream signalling pathways through G-protein-coupled receptors to achieve numerous functions on oocyte maturation and embryo development.In this study,sheep in vitro fertilized embryos were applied to investigate the effects of LPA on early embryos development and embryonic stem cell establishment.At first,the maturation medium containing estrus female sheep serum and synthetic oviduct fluid(SOF)were optimized for sheep IVF,and then the effects of LPA were investigated.From 0.1 to 10μmol L^(–1),LPA had no significant effect on the cleavage rate(P>0.05),but the maturation rate and blastocyst rate increased dependently with LPA concentration(P<0.05),and the blastocyst morphology was normal.When the LPA concentration was 15μmol L^(–1),the maturation rate,cleavage rate and blastocyst rate decreased significantly(P<0.05),and the blastocyst exhibited abnormal morphology and could not develop into highquality blastocyst.Besides,the exogenous LPA increases the expression of LPAR2,LPAR4,TE-related gene CDX-2and pluripotency-related gene OCT-4 in sheep early IVF embryos with the raise of LPA concentration from 0.1 to 10μmol L^(–1).The expression of LPAR2,LPAR4,CDX-2 and OCT-4 from the LPA-0.1μmol L^(–1)to LPA-10μmol L^(–1)groups in early embryos were extremely significant(P<0.05),while the expression of these genes significantly decreased in 15μmol L^(–1)LPA-treated embryos compared with LPA-10μmol L^(–1)group(P<0.05).The inner cell mass in 15μmol L^(–1)LPA-treated embryos was also disturbed,and the blastocysts formation was abnormal.Secondly,the sheep IVF blastocysts were applied to establish embryonic stem cells.The results showed that LPA made the blastocyst inoculated cells grow towards TSC-like cells.They enhanced the fluorescence intensity and mRNA abundance of OCT-4 and CDX-2 as the concentration increased from 0 to 10μmol L^(–1),while 15μmol L^(–1)LPA decreased OCT-4 and CDX-2 expression in the derived cells.The expression of CDX-2 and OCT-4 in the blastocyst inoculated cells of LPA-1μmol L^(–1)group and LPA-10μmol L^(–1)group extremely significantly increased(P<0.05),but there was significant decrease in LPA-15μmol L^(–1)group compared with LPA-10μmol L^(–1)group(P<0.05).Meanwhile,the protein expression of LPAR2 and LPAR4 remarkably increased after treatment of LPA at 10μmol L^(–1)concentration.This study references the IVF embryo production and embryonic stem cell research of domestic animals.

The barriers of joining in vitro fertilization programs among infertile couples in developing countries:A scoping review

Objective:To determine the barriers of joining in-vitro fertilization(IVF)programs among infertile couples in developing countries.Methods:This study assessed infertile couples and the barriers or associated factors resulting in delayed decision-making of joining IVF program by searching databases PubMed,ScienceDirect,EBSCO,ClinicalKey,and Cochrane Library from inception until December 31,2021.Additional search strategies were snowballing literature search and citation tracking.Results:Eleven articles were included in the scoping review.The cost was the greatest barrier of joining IVF program.Limited access and lack of assisted reproductive technology centers,few qualified infertility trained staff,insufficient government support,low priority in government policy,along with sociocultural factors,such as religion and false beliefs or myths were also majorly considered to be associated obstacles.Conclusions:The main barrier associated with IVF program among infertile couples in developing countries is the high cost of the IVF services.

Live births from in vitro fertilization-embryo transfer following the administration of gonadotropin-releasing hormone agonist without gonadotropins:Two case reports

BACKGROUND The prevalence of female infertility between the ages of 25 and 44 is 3.5%to 16.7%in developed countries and 6.9%to 9.3%in developing countries.This means that infertility affects one in six couples and is recognized by the World Health Organization as the fifth most serious global disability.The International Committee for Monitoring Assisted Reproductive Technology reported that the global total of babies born as a result of assisted reproductive technology procedures and other advanced fertility treatments is more than 8 million.Advancements in controlled ovarian hyperstimulation procedures led to crucial accomplishments in human fertility treatments.The European Society for Human Reproduction and Embryology guideline on ovarian stimulation gave us valuable evidence-based recommendations to optimize ovarian stimulation in assisted reproductive technology.Conventional ovarian stimulation protocols for in vitro fertilization(IVF)–embryo transfer are based upon the administration of gonadotropins combined with gonadotropin-releasing hormone(GnRH)analogues,either GnRH agonists(GnRHa)or antagonists.The development of ovarian cysts requires the combination of GnRHa and gonadotropins for controlled ovarian hyperstimulation.However,in rare cases patients may develop an ovarian hyper response after administration of GnRHa alone.CASE SUMMARY Here,two case studies were conducted.In the first case,a 33-year-old female diagnosed with polycystic ovary syndrome presented for her first IVF cycle at our reproductive center.Fourteen days after triptorelin acetate was administrated(day 18 of her menstrual cycle),bilateral ovaries presented polycystic manifestations.The patient was given 5000 IU of human chorionic gonadotropin.Twenty-two oocytes were obtained,and eight embryos formed.Two blastospheres were transferred in the frozen-thawed embryo transfer cycle,and the patient was impregnated.In the second case,a 37-year-old woman presented to the reproductive center for her first donor IVF cycle.Fourteen days after GnRHa administration,the transvaginal ultrasound revealed six follicles measuring 17-26 mm in the bilateral ovaries.The patient was given 10000 IU of human chorionic gonadotropin.Three oocytes were obtained,and three embryos formed.Two high-grade embryos were transferred in the frozen-thawed embryo transfer cycle,and the patient was impregnated.CONCLUSION These two special cases provide valuable knowledge through our experience.We hypothesize that oocyte retrieval can be an alternative to cycle cancellation in these conditions.Considering the high progesterone level in most cases of this situation,we advocate freezing embryos after oocyte retrieval rather than fresh embryo transfer.

Early embryonic failure caused by a novel mutation in the TUBB8 gene:A case report

BACKGROUND This study aimed to explore the relationship between gene mutations and early embryonic development arrest and to provide more possibilities for the diagnosis and treatment of repeated implantation failure.CASE SUMMARY Here,we collected and described the clinical data of a patient with early embryonic development stagnation after repeated in vitro fertilization attempts for primary infertility at the Department Reproductive Center of Zaozhuang Maternal and Child Healthcare Hospital.We also detected the whole-exon gene of the patient's spouse and parents,and conducted bioinformatics analysis to determine the pathogenesis of the gene.CONCLUSION A novel mutant of the TUBB8 gene[c.602G>T(p.C201F)]was identified,and this mutant provided new data on the genotype-phenotype relationships of related diseases.

Effect of Mitochondrial Function of Ovarian Granulosa Cells on In Vitro Fertilization and Embryo Transfer Outcomes in Obese Polycystic Ovary Syndrome Patients

Objective:To investigate the effect of abnormal ovarian granulosa cell metabolism on in vitro fertilization and embryo transfer(IVF-ET)outcomes in obese polycystic ovary syndrome(PCOS)patients.Methods:Patients with PCOS who met the study criteria were screened according to the inclusion criteria.A total of 32 patients with obese PCOS were recruited into the study group,and 39 patients with non-obese PCOS were recruited into the control group.The general data(age,body mass index,and years of infertility),insulin resistance index(HOMA-IR),follicle-stimulating hormone(FSH),luteinizing hormone(LH),granulosa cell mitochondrial function,and IVF-ET outcome of patients in the study group and control group were retrospectively analyzed.Results:The differences in age and years of infertility between the study group and the control group were insignificant(P>0.05),and the body mass index(BMI)of the study group and control group was 30.5±1.24 kg/m2 and 22.3±1.12 kg/m2,respectively,in which the difference was statistically significant(P<0.05);the HOMA-IR of the study group was significantly higher than that of the control group(P<0.05);the reactive oxygen species(ROS)in the study group was significantly higher than that in the control group(P<0.05),and the ATP content in the study group was significantly lower than that in the control group(P<0.05);comparing the FSH and LH levels between the two groups,the difference was not statistically significant(P>0.05);the rate of IVF-ET failure was significantly higher in the study group than in the control group.Conclusion:PCOS is a complex endocrine disorder,and obesity is one of the independent risk factors for the development of PCOS.

Clinical Effects of the Follicular Phase Long Regimen and Luteal Phase Long Regimen on Ovulation Induction in IVF-ET Treatment: A Meta-Analysis

Objective:To systematically evaluate the clinical effects of the follicular phase long regimen and the luteal phase long regimen on ovulation induction in IVF-ET treatment.Methods:Databases including PubMed,Embase,Cochrane Library,CNKI,Chinese Biomedical Literature(CBM),VIP,Wanfang,and others were searched up to January 2021.Clinical studies on ovulation induction using the follicular phase long regimen and luteal phase long regimen in IVF-ET treatment were identified.Literature screening,data extraction,and quality evaluation were conducted based on inclusion and exclusion criteria.Meta-analysis was performed using RevMan 5.3 software.Results:After screening,a total of 11 studies were included,comprising 21,544 patients:9,974 in the follicular phase long regimen group and 11,570 in the luteal phase long regimen group.The meta-analysis results were as follows:(1)The number of Gn days and the total amount of Gn in the follicular phase long regimen were higher than those in the luteal phase long regimen(P<0.05);(2)The number of eggs obtained in the follicular phase long regimen was higher than that in the luteal phase long regimen(P<0.05).There were no significant differences in the rate of embryo optimization and cycle cancellation between the two groups(P>0.05);(3)The embryo implantation rate and clinical pregnancy rate in the follicular phase long regimen were higher than those in the luteal phase long regimen(P<0.05),while the abortion rate in the follicular phase long regimen was lower than that in the luteal phase long regimen(P<0.05).Conclusion:Compared to the luteal phase long regimen,the follicular phase long regimen involves more Gn days and a higher total amount of Gn.The optimal embryo rate and cycle cancellation rate were similar between the regimens,but the follicular phase long regimen resulted in more eggs,significantly improved the implantation and clinical pregnancy rates,and reduced the abortion rate.However,these conclusions require further validation through more multicenter,large-sample RCT studies.

Intracytoplasmic sperm injection in cases with a history of in vitro fertilization failure

<abstract>Aim: To evaluate the effect of intracytoplasmic sperm injection (ICSI) in the management of cases with a history of conventional in vitro fertilization (IVF) failure. Methods: Two groups of patients, 19 with normal semen parameters and a history of IVF failure (metaphase Ⅱ oocytes: 0~30 %) and 28 with severe male factor infertility received ICSI technology during the same period. Ovarian stimulation was achieved by conventional procedure. Transvaginal ultrasound-guided oocyte collection was done 35~37 h after human chorionic gonadotrophin (hCG) injection. Only metaphase Ⅱ oocytes were selected for microinjection. Results: Fertilization was achieved with ICSI in all the patients. The fertilization rate (75.6 %±21.1 % vs. 73.9 %±19.2 %), cleavage rate (85.1 %±19.3 % vs. 82.7 %±22.1 %), clinical pregnancy rate per embryo transfer cycle (31.6 % vs. 28.6 %) and implantation rate per embryo (15.3 % vs. 14.4 %) did not differ significantly between the two groups. Conclusion: ICSI is a valuable method for couples with a history of IVF failure. These patients may have a similar ICSI result as in severe male infertility.

Untreated Prior Pulmonary Tuberculosis Adversely Affects Pregnancy Outcomes in Infertile Women Undergoing in vitro Fertilization and Embryo Transfer: A Large Retrospective Cohort Study

Objective Prior pulmonary tuberculosis(PTB) on chest X-ray(CXR) was commonly found in infertile patients receiving examinations before in vitro fertilization and embryo transfer(IVF-ET). It was unclear whether untreated PTB would affect pregnancy outcomes after IVF-ET.Method We conducted a retrospective cohort study of 14,254 infertile patients who had received IVFET at Peking University Third Hospital in 2017. Prior PTB was defined as the presence of signs suggestive of old or inactive PTB on CXR, with or without a clinical TB history. Patients who had prior PTB on CXR but had not received a clinical diagnosis and anti-TB therapy were included for analysis. Live birth,clinical pregnancy, and miscarriage rates were compared between the untreated PTB and non-PTB groups.Results The untreated PTB group had significantly lower clinical pregnancy(31.7% vs. 38.1%) and live birth(23.8% vs. 30.6%) rates than the non-PTB group(both P < 0.001). Multivariate analysis revealed that untreated PTB was a risk factor for decreased live birth rate [odds ratio(OR), 0.80;95% confidence interval(CI), 0.66–0.98;P = 0.028] in all patients and for increased miscarriage(OR, 4.19;95% CI,1.69–10.39;P = 0.002) and decreased live birth(OR, 0.45;95% CI, 0.24–0.83;P = 0.011) rates in patients with unexplained infertility.Conclusions Untreated PTB was associated with adverse pregnancy outcomes after IVF-ET, especially in patients with unexplained infertility, highlighting the clinical significance of PTB in this specific patient population.

Reproductive Outcomes in Women with Prior Cesarean Section Undergoing In Vitro Fertilization:A Retrospective Case-control Study

The impact of prior cesarean section（CS） on the pregnancy and neonatal outcomes of in vitro fertilization and embryo transfer（IVF-ET） was investigated. A retrospective analysis was performed on 144 patients with prior CS between January 2013 and December 2015. The pregnancy, delivery, and neonatal outcomes of patients who had previous CS delivery and received IVF-ET were analyzed. The control group comprised 166 patients who had only previous vaginal delivery（VD） and received IVF-ET during the same period. The results showed that the basal follicle stimulating hormone level, estradiol level on human chorionic gonadotropin（h CG） day, gonadotrophin dosage, duration of stimulation, retrieved oocytes, fertilization rate, high-quality embryo rate, multiple birth rate, abortion rate and ectopic pregnancy rate had no significant difference between the two groups（P〉0.05）. The pregnancy rate（40.28% vs. 54.22%） and implantation rate（24.01% vs. 34.67%） were significantly lower（P〈0.05）, and the ratio of embryo difficulty transfer（9/144 vs. 0/166） was significantly higher in CS group than in VD group. The risk of pernicious placenta previa and postpartum hemorrhage in twin deliveries was significantly increased in CS group as compared with that in VD group（P〈0.05）, and gestational age and neonatal birth weight were significantly reduced in twin deliveries as compared with singleton deliveries in both groups（P〈0.05）. It was suggested that the existence of CS scar may impact embryo implantation and clinical pregnancy outcome, and increase the difficulty of ET. We should limit the number of transfer embryos to avoid multiple pregnancies and strengthen gestational supervision in patients with cesarean scar.

Does Lower Dose of Long-acting Triptorelin Maintain Pituitary Suppression and Produce Good Live Birth Rate in Long Down-regulation Protocol for In-vitro Fertilization?

The effects of pituitary suppression with one-third depot of long-acting gonadotropin-releasing hormone（Gn RH） agonist in Gn RH agonist long protocol for in vitro fertilization（IVF）/intracytoplasmic sperm injection（ICSI） were investigated. A retrospective cohort study was performed on 3186 cycles undergoing IVF/ICSI with Gn RH agonist long protocol in a university-affiliated infertility center. The pituitary was suppressed with depot triptorelin of 1.25 mg or 1.875 mg. There was no significant difference in live birth rate between 1.25 mg triptorelin group and 1.875 mg triptorelin group（41.2% vs. 43.7%）. The mean luteinizing hormone（LH） level on follicle-stimulating hormone（FSH） starting day was significantly higher in 1.25 mg triptorelin group. The mean LH level on the day of human chorionic gonadotrophin（h CG） administration was slightly but statistically higher in 1.25 mg triptorelin group. There was no significant difference in the total FSH dose between the two groups. The number of retrieved oocytes was slightly but statistically less in 1.25 mg triptorelin group than in 1.875 mg triptorelin group（12.90±5.82 vs. 13.52±6.97）. There was no significant difference in clinical pregnancy rate between the two groups（50.5% vs. 54.5%）. It was suggested that one-third depot triptorelin can achieve satisfactory pituitary suppression and produce good live birth rates in a long protocol for IVF/ICSI.

Intramural pregnancy after in vitro fertilization and embryo transfer:A case report

BACKGROUND Intramural pregnancy is a rare type of ectopic pregnancy,which is diagnosed by transvaginal ultrasound and magnetic resonance imaging.Management strategies vary depending on the site of the pregnancy,the gestational age and the desire to maintain fertility.The incidence of intramural pregnancy in assisted reproductive technology is higher than that in natural pregnancy.CASE SUMMARY We present a case of intramural pregnancy after in vitro fertilization and elective single embryo transfer following salpingectomy.The patient was completely asymptomatic and her serumβ-human chorionic gonadotropin level increased from 290 m IU/m L to 1759 m IU/m L.Three-dimensional transvaginal ultrasound indicated a heterogeneous echogenic mass arising from the uterine fundus which was surrounded by myometrium and a slender and extremely hypoechoic area stretching to the uterine cavity which was thought to be a fistulous tract.Therefore,we considered a diagnosis of intramural pregnancy and laparoscopic surgery was conducted at 7 wk gestation.CONCLUSION Early diagnosis and treatment of intramural pregnancy is significant for maintaining fertility.

A nomogram predicting clinical pregnancy in the first fresh embryo transfer for women undergoing in vitro fertilization and intracytoplasmic sperm injection(IVF/ICSI) treatments

The extent to which factors affect the probability of clinical pregnancy in the first fresh embryo transfer after assisted conception is unknown.In order to examine the predictors of clinical pregnancy,a retrospective cohort study was launched between January 1,2013 and December 31,2016 in four infertility clinics including 19837 in vitro fertilization and intracytoplasmic sperm injection(IVF/ICSI)fresh cycles with known outcomes and relevant records.A multivariable logistic regression was used to select the most significant predictors in the final nomogram for predicting clinical pregnancy.Furthermore,the model was validated by an independent validation set and the performance of the model was evaluated by the receiver operating characteristic(ROC)curves along with the area under the ROC curve(AUC)and calibration plots.In a training set including 17854 participants,we identified that female age,tubal factor,number of embryos transferred,endometrial thickness and number of good-quality embryos were independent predictors for clinical pregnancy.We developed a nomogram using these five factors and the predictive ability was 0.66 for AUC(95%CI=0.64−0.68),which was independently validated in the validation set(AUC=0.66,95%CI=0.65−0.68).Our results show that some specific factors can be used to provide infertile couples with an accurate assessment of clinical pregnancy following assisted conception and facilitate to guide couples and clinicians.

A highly homozygous and parthenogenetic human embryonic stem cell line derived from a one-pronuclear oocyte following in vitro fertilization procedure

因为他们的人的白血球抗原(HLA ) haplotype 将强烈增加与房间线的相对更小的队为某些人口匹配的度，同型结合的人的胚胎的干细胞(hESCs ) 被认为是为 hESC 银行的更好的房间来源。同型结合的 hESCs 能从单性生殖的胚胎被产生，但是仅仅异质接合的 hESCs 用当前的策略被建立了没有第二极的身体挤出，人工地激活卵母细胞。这里，我们在 vitro 授精处理从 one-pronuclear 卵母细胞追随者例程报导一根人的同型结合的转换字符线(chHES-32 ) 的第一成功的推导。chHES-32 房间与正常 hESCs 表示普通标记和基因。他们处于一个无差别的状态被宣传超过一年(】P50 ) 了并且维持了 46 的稳定的 karyotype， XX。当在 vivo 并且在 vitro 区分了时， chHES-32 房间能从所有三胚胎的细菌层形成衍生物。五父亲一般地表示的印的基因和他们与卵母细胞施主相同的 HLA 遗传型的几乎无法发现的表示显示了他们的单性生殖的起源。用染色体宽的单个核苷酸的多型性分析并且 DNA 采指纹， chHES-32 房间的 homozygosity 进一步被证实。结果显示“讨厌”的 one-pronuclear 卵母细胞可能是为人的同型结合、单性生殖的转换字符的潜在的来源，并且为从单性生殖的 haploid 卵母细胞获得同型结合的 hESC 线建议了其他的策略。

Effects of Ovarian Preservation Time on in vitro Maturation and in vitro Fertilization of Oocytes from Slaughtered Sheep

[Objectives] The aim was to study the effects of ovarian preservation time on in vitro fertilization of oocytes from slaughtered sheep. [Methods] The collected ovaries were randomly and evenly divided into four groups. They were preserved in physiological saline containing penicillin（ 100 IU/ml） and streptomycin（ 100 μg/ml） at 15-20 ℃ for 0（ Control）,6,12 and 18 h,respectively. Then,the oocytes were subjected to in vitro fertilization. [Results]The maturation rates,cleavage rates and blastocyst rates of the oocytes preserved for 6 and 12 h showed no significant differences compared with those of the oocytes preserved for0 h（ 72. 03%,70. 87% vs. 73. 68%; 74. 12%,72. 60% vs. 74. 49%; 22. 22%,20. 75% vs. 23. 29%）（ P 〉 0. 05）. There were also no significant differences in maturation rate,cleavage rate or blastocyst rate between the oocytes preserved for 18 and 0 h（ P 〉 0. 05）. [Conclusions] Within a certain rage（ 0-18 h）,storage time of ovary at 15-20 ℃ does not affect the continued development of oocytes from slaughtered sheep.

In Vitro Fertilization Outcomes Following Assisted Hatching of Embryos with Thick Zona Pellucida—A Prospective Randomized Study

Purpose: Impaired hatching is associated with implantation failure following in vitro fertilization (IVF). Thickening or hardening of the zona pellucida (ZP) has been proposed as a factor in this impairment. We examined whether selective assisted hatching (AH) is beneficial with embryos having a thick ZP. Methods: This prospective, randomized controlled study was performed in the IVF unit of an obstetrics and gynecology department in a university-affiliated hospital. Only patients undergoing IVF and having a ZP thickness of ≥17 μm measured in all their embryos were included. In the intervention group, AH was applied to all embryos, before their transfer. In the control group, embryos were transferred without AH. Implantation, clinical pregnancy and live birth rates were the study endpoints. Results: Both study arms were comparable in most baseline parameters. The two groups did not differ in implantation rates (14.1% control vs. 8.92% intervention, odds ratio (OR) = 0.5974, 95% confidence interval (CI) 0.325 - 1.1), clinical pregnancy rates (36.7% vs. 25.8%, OR = 0.6025, 95% CI 0.274 - 1.325), or live birth rates (25% vs. 18.9%, OR = 0.7021, 95% CI 0.291 - 1.691). Conclusions: Selecting embryos for AH by their ZP thickness as a sole parameter was not found to be beneficial and to improve IVF outcome.

A novel in vitro system for gamete fusion in maize

Various systems by using electric pulse, calcium, or polyethylene glycol have been developed in the past decade for the in vitro fusion of plant gametes. These in vitro systems provide a new way to study the fertilization mechanisms of plants. In this study, we developed a bovine serum albumin (BSA)-mediated fusion system for the in vitro fusion of maize gametes. The in vitro fusion of the isolated single egg cell and sperm cell of maize was observed microscopically in the BSA solution and the fertilized egg cell showed normal cell wall regeneration and nuclear division. The effects of the BSA concentration, pH value and calcium level on the efficiency of the maize gamete fusion were also assessed. BSA concentration and pH value did significantly affect the efficiency of the gamete fusion. Calcium was not necessary for the gamete fusion when BSA was present. The optimal solution for the gamete fusion contained 0.1% BSA, pH 6.0. The fusion frequency was as high as 96.7% in that optimal solution. This new in vitro fertilization system offers an alterna- tive tool for the in vitro study of fertilization mechanisms with much simpler manipulating procedure than PEG system, and it will be especially useful for the in vitro study of the calcium dynamics during plant fertilization.

Post-salpingectomy interstitial heterotopic pregnancy after in vitro fertilization and embryo transfer:A case report

BACKGROUND Heterotopic pregnancy(HP)refers to the coexistence of ectopic pregnancy and intrauterine pregnancy.Salpingectomy is proposed as a pretreatment before in vitro fertilization and embryo transfer(IVF-ET)to reduce the risk of HP.HP after IVF-ET occurs in women who had already underwent bilateral salpingectomy,even though it is extremely rare.CASE SUMMARY A case of a 29-year-old woman with recurrent interstitial HP after IVF-ET following salpingectomy is presented.The main symptom was a sudden and worsening pelvic pain.Physical examinations revealed signs of peritoneal bleeding and irritation with stable vital signs.Transvaginal ultrasound showed a live intrauterine pregnancy and another live embryo with cardiac activity in the left cornu extending beyond the lateral edge of the uterus.Her hemoglobin concentration was 8.0 g/dL,and serum human chorionic gonadotropin value was 171116.9 mIU/mL.With the diagnosis of ruptured HP with internal bleeding,an emergency laparoscopic resection of left cornu was performed.The interstitial pregnancy was removed with caution to protect the intrauterine pregnancy.After the surgical treatment,the intrauterine pregnancy continued with no complications.A healthy baby was delivered by caesarean section at 39 wk.Outcomes of another three cases are further summarized.CONCLUSION Post-salpingectomy HP is a rare but challenging condition.Surgical treatment is preferred in the case with a viable intrauterine pregnancy.

Mesenteric vein thrombosis following impregnation via in vitro fertilization-embryo transfer

Pregnancy is an acquired hypercoagulable state. Most patients with thrombosis that develops during pregnancy present with deep vein leg thrombosis and/or pulmonary embolism, whereas the development of mesenteric vein thrombosis(MVT) in pregnant patients is rare. We report a case of MVT in a 34-year-old woman who had achieved pregnancy via in vitro fertilization-embryo transfer(IVFET). At 7 wk of gestation, the patient was referred to us due to abdominal pain accompanied by vomiting and hematochezia, and she was diagnosed with superior MVT. Following resection of the gangrenous portion of the small intestine, anticoagulation therapy with unfractionated heparin and thrombolysis therapy via a catheter placed in the superior mesenteric artery were performed, and the patient underwent an artificial abortion. Oral estrogen had been administered for hormone replacement as part of the IVF-ET procedure, and additional precipitating factors related to thrombosis were not found. Pregnancy itself, in addition to the administered estrogen, may have caused MVT in this case. We believe that MVT should be included in the differential diagnosis of a pregnant patient who presents with an acute abdomen.

Comparison between in vitro fertilization outcome of two motile sperm separation methods

Objective: To evaluate the efficiency of Percoll density gradient and swim-up methods for motile sperm isolation for in vitro fertilization and embryo transfer (IVF-ET) program. Methods: The fertilization rate, cleavage rate, embryo developing status and pregnancy outcome of 362 IVF cycles using sperm obtained by the two methods were studied. Results: There was no significant difference in fertilization rate and cleavage rate between the Percoll and swim-up groups. Although the two groups showed no significant difference in the embryo cell number, the percentage of embryos with<20% debris was significantly higher in the Percoll group (77.6%) than in swim-up group (65.9%). The pregnancy rate and the life birth rate were also significantly higher (P<0.01) in the Percoll group (43.7% and 70.3%, respectively) than in the swim-up group (36.6 % and 60.7 %, respectively). Conclusion: The efficiency of the Percoll density gradient method is superior to the swim-up method in motile sperm separation for the IVF-ET program. [Reprod Contracep (in Chinese) 2002; 22: 292]