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.

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.

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.

Awareness and Attitudes Toward In Vitro Fertilization in Patients with Infertility:A Cross-Sectional Study

Objective:To investigate the awareness and attitudes of patients with infertility toward in vitro fertilization(IVF).Methods:A cross-sectional study was conducted in patients with infertility at the Reproductive Medicine Center of Peking University Third Hospital.Data collection was conducted using a questionnaire that consists of patient background information,methods of obtaining relevant IVF information,and patients’awareness of IVF treatment as well as the law and ethics.Results:A total of 198 participants of different backgrounds were enrolled in the study,and IVF information was obtained through different means;however,most of the participants had a lack of awareness and improper attitude toward IVF.The participants had higher estimations of treatment expenses(Pearson λ^(2)=22.132,P<0.05)and time(Pearson λ^(2)=47.992,P<0.05)during the initial consultation,while those with treatment experience expressed fewer concerns about IVF therapy(Pearson λ^(2)=9.467,P<0.05),particularly about the genetic risks of having IVF.Conclusion:Measures should be taken to improve patients’awareness and attitudes toward IVF,and a large-scale and multicenter clinical study should be conducted in the future.

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.

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.

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.

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.

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.

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]

Effects of difructose dianhydride (DFA)-Ⅳ on in vitro fertilization in pigs

Difructose dianhydride Ⅳ(DFA-Ⅳ) is produced from levan, which is a natural polysaccharide that belongs to the fructan family, through the activity of levan fructotransferase(LF) derived from microorganisms. Recently, DFA-Ⅳ has been expected to have diverse applications in the food and medical industry. Here, we examined the potential application of DFA-Ⅳ for in vitro fertilization(ⅣF) in pigs. In the assessment of acrosomal integrity during incubation, intact acrosomal or viable spermatozoa were highly sustained in 0.1% or 0.25% DFA-Ⅳ(69.8%-70.8%,P<0.05). Reactive oxygen species(ROS) levels during sperm incubation decreased following the addition of DFA-Ⅳ, and 0.1 %-0.5% DFA-Ⅳ in particular significantly decreased ROS production relative to that seen with no addition or 0.75% DFA-Ⅳ. Total fertilization(mono + polyspermic oocyte) rate was significantly higher in the addition of0.1% DFA-Ⅳ(94.2%) than with other concentrations(71.8%-86.7%, P<0.05). When using reduced ⅣF times and lower sperm numbers, we found that addition of 0.1%-0.5% DFA-Ⅳ significantly increased the fertilization rate(P<0.05). Fertilized oocytes treated with 0.1% DFA-Ⅳ exhibited higher embryonic development and blastocyst formation than those treated with other concentrations(P<0.05). Consequently, the addition of DFA-Ⅳ during ⅣF improved fertilization and embryonic development, suggesting the possible use of novel sugars for enhancement of assisted reproductive technology(ART) in mammals.

Effect of royal jelly on in vitro fertilization and early embryo development following nicotine treatment in adult female rats

Objective:To scrutinize the protective role of royal jelly as an antioxidant on nicotine-induced changes in malondialdehyde(MDA)level,p53 expression,in vitro fertilization(IVF)rate,and early embryo development in adult female rats.Methods:A total of 56 adult female Wistar rats were divided into 8 groups(n=7 in each group).Group 1 served as an untreated control group,group 2,3 and 4 received nicotine at a dose of 0.50,1.00 and 2.00 mg/kg respectively,group 5 received royal jelly at a dose of 100.00 mg/kg,and group 6,7 and 8 received 0.50,1.00 and 2.00 mg/kg nicotine,respectively,with 100.00 mg/kg body weight royal jelly.Nicotine and royal jelly were administered daily for 49 days in the experimental groups intra-peritoneally and orally,respectively.At the end of the experimental period,p53 expression,IVF rate and early embryo development as well as MDA concentration were measured.Results:The IVF rate,number of cumulus oocytes,two-cell embryos and blastocysts decreased in the nicotine-treated groups in a dose-dependent manner.In addition,p53 mRNA expression and MDA levels increased in the nicotine-treated groups.Royal jelly co-administration led to partial improvement in the aforementioned parameters.Conclusions:Royal jelly may have a repro-protective effect in nicotine-administered female rats in terms of its anti-oxidant and anti-apoptotic properties.

Evaluation of pig oocyte in vitro maturation and fertilization using three gonadotropin-based hormonal compounds

Objective:To evaluate the effect of human chorionic gonadotropins(hCG)and equine chorionic gonadotropins(eCG)on in vitro gilt oocyte maturation and embryonic development,using frozen semen for fertilization.Methods:Two independent experiments(6 replicates each)were carried out to evaluate gilt oocyte maturation,and fertilization and embryonic development by using ovaries from a local abattoir.Totally,712 oocytes were randomly distributed in four-well dishes to receive Novormon(eCG 5.0 IU),PG600(eCG 5.0 IU and hCG 2.5 IU),Chorulon(hCG 5.0 IU),or no hormones.Oocytes were incubated with 5%CO2,95%air and saturation humidity at 39℃for 44 h.Maturation of the oocytes to metaphaseⅡwas assessed by using the aceto-orcein technique.In addition,741 oocytes were used and randomly distributed in four-well dishes,and then oocyte maturation was carried out as mentioned,but matured oocytes were washed and placed in fertilization medium with frozen-thawed sperm.Gametes were co-incubated for 7 h,and then washed and placed in development medium,and incubated for further 7 days,at which time embryonic development was evaluated.Fertilization and embryo development media were not supplemented with the studied hormones.Results:Novormon(eCG)and PG600(eCG+hCG)treatments significantly improved the percentages of metaphaseⅡoocytes compared to the control group(P<0.05).Furthermore,a significant increase was also observed in the young blastocyst stage between the control group and the PG600 treatment group(P<0.05).Conclusions:Hormonal products Novormon(eCG)and PG600(eCG+hCG)can obtain the highest percentages of in vitro maturation in gilt oocytes;however,this effect is not transferred to fertilization rates.

Effects of Different Culture Systems on In Vitro Fertilization of Oocyte in Bovine

[ Objective] To investigate the optimal culture system for in vitro fertilization （IVF） of oocyte in bovine. [ Method] The IVF of mature bovine oocytes was conducted and the fertilized eggs were cultured in vitro with different culture systems. In the traditional culture system, the TALP medium and BO medium were used as fertilization medium, respectively; and M-199 medium supplemented with 5% FCS was used as fertilized egg culture medium. In the transitional culture system, the TALP medium and BO medium were used as fertilization medium; 60% TALP and 40% M-199 supplemented with 5% FCS and 60% BO and 40% M-199 supplemented with 5% FCS were used as fertilized egg culture medium, respectively. The effects of different culture systems on fertilization rate and cleavage rate were observed. E Result] In the transitional culture system （with TALP medium as fertilization medium and with 60% TALP and 40% M-199 supplemented with 5% FCS as fertilized egg culture medium）, the fertilization rate and cleavage rate were increased to 77.8% and 55.6%, respectively. [ Conclusion] The transitional culture system （with TALP medium as fertilization medium and with 60% TALP and 40% M-199 supplemented with 5% FCS as fertilized egg culture medium） is the optimal culture system for IVF of bovine oocyte.

Ectopic pregnancy and failed oocyte retrieval during in vitro fertilization stimulation:Two case reports

BACKGROUND Due to a slight rise in beta-human chorionic(β-h CG)levels that are undetectable,and vaginal bleeding that is similar to regular menstruation,ectopic pregnancy(EP)that occurs during the expected menstrual cycle prior to ovulation induction as part of in vitro fertilization(IVF)is likely to be undiagnosed.We present two cases of unexpected EP and emphasize the importance of theβ-h CG assay when an unexplained increase in progesterone is present prior to the triggering of ovulation during controlled ovarian stimulation(COS).CASE SUMMARY A 26-year-old woman with primary infertility and a 31-year-old woman with secondary infertility.Both patients sought IVF treatment due to fallopian tube obstruction and underwent COS using the gonadotropin-releasing-hormone(Gn RH)-antagonist protocol.In the late stage of COS,progesterone levels in both patients significantly increased,and luteinizing hormone levels decreased,followed by oocyte retrieval failure.A right salpingectomy was performed and tubal ectopic pregnancy was diagnosed by pathology in the first patient,and the second patients was diagnosed with a suspected EP abortion because herβ-h CG levels declined to 12.5 m IU/m L.After full recovery for 2 mo,the first patient entered a new IVF treatment cycle with a Gn RH-antagonist regimen and successfully achieved eight oocytes and three viable embryos.After 6 mo,the second patient received another COS treatment with a progestin-primed ovarian stimulation protocol and successfully achieved nine oocytes and five viable embryos.CONCLUSIONβ-h CG levels in the initial and midterm phases of COS must be considered in patients with unusual hormone dynamics.