Kidding after Transfer of in Vitro Produced Saanen Goat Embryos into Local Ukrainian Breed Recipients in Different Seasons

In recent years, the demand for goat products has been growing due to the fact that goat milk has a number of advantages over cow milk, for example, it is low in lactose, and is considered less allergenic and easier to digest. To increase production during both breeding and non-breeding seasons and reduce the price of dairy products, it is necessary to effectively use reproductive management and assisted reproductive technologies. In vitro embryo production makes it possible to obtain a large number of eggs from goats, which for some reason are unable to conceive, but have genetic value. Afterward in vitro produced embryos can be transferred into recipient goats of other less genetically valuable breeds, such as the Ukrainian local breed. Therefore, the aim of the present study was to investigate the effectiveness of transfers of in vitro produced embryos of Saanen goats into surrogate sires of the Ukrainian local breed in different seasons. All manipulations with animals were carried out following ethical standards (Strasbourg, 1986). Six Saanen goats were selected as the oocyte donors. After the hormonal stimulation oocytes were retrieved by laparoscopic ovum pick-up. In vitro produced embryos were transferred laparotomically into 24 recipients of Ukrainian local breed. Fifty days after embryo transfers, pregnancies were determined by ultrasound diagnostics. Although the embryo development rate in the breeding season was 20% higher than in the non-breeding season, there was no difference in pregnancy and kidding rates between seasons. In conclusion, the transfer of in vitro produced Saanen goat embryos to recipients of the Ukrainian local breed gives the opportunity to achieve pregnancy and kidding regardless of the breeding season, which will enable a faster and more efficient increase in the livestock of highly productive goats in Ukraine in the post-war period.

Reproductive fluids,used for the in vitro production of pig embryos,result in healthy offspring and avoid aberrant placental expression of PEG3 and LUM

Background:In vitro embryo production(IVP)and embryo transfer(ET)are two very common assisted reproductive technologies(ART)in human and cattle.However,in pig,the combination of either procedures,or even their use separately,is still considered suboptimal due to the low efficiency of IVP plus the difficulty of performing ET in the long and contorted uterus of the sow.In addition,the potential impact of these two ART on the health of the offspring is unknown.We investigated here if the use of a modified IVP system,with natural reproductive fluids(RF)as supplements to the culture media,combined with a minimally invasive surgery to perform ET,affects the output of the own IVP system as well as the reproductive performance of the mother and placental molecular traits.Results:The blastocyst rates obtained by both in vitro systems,conventional(C-IVP)and modified(RF-IVP),were similar.Pregnancy and farrowing rates were also similar.However,when compared to in vivo control(artificial insemination,AI),litter sizes of both IVP groups were lower,while placental efficiency was higher in AI than in RF-IVP.Gene expression studies revealed aberrant expression levels for PEG3 and LUM in placental tissue for C-IVP group when compared to AI,but not for RF-IVP group.Conclusions:The use of reproductive fluids as additives for the culture media in pig IVP does not improve reproductive performance of recipient mothers but could mitigate the impact of artificial procedures in the offspring.

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.

Supplementation of L-ascorbic acid improves the in vitro development of buffalo(Bubalus bubalis)embryos and alters the expression of apoptosis-related genes

Objective:To study the effect of L-ascorbic acid supplementation on the in vitro development of buffalo embryos and evaluate the relative mRNA abundance of some pro-apoptotic,anti-apoptotic,and embryonic development-related genes.Methods:In experiment 1,we evaluated the effect of the addition of 0(control),50,and 100μM L-ascorbic acid to the in vitro maturation medium on the developmental competence in terms of blastocyst rate and relative mRNA abundance of some pro-apoptotic(BAX,BID),anti-apoptotic(BCL-XL,MCL1),and embryonic development(GDF9,BMP15)related genes.Based on the results,we chose 50μM as the suitable dose of L-ascorbic acid for the subsequent experiments.We further evaluated the blastocyst rates following the addition of 50μM L-ascorbic acid to the in vitro culture medium(experiment 2),and in vitro maturation and in vitro culture media(experiment 3).In all three experiments,the maturation and culture media devoid of L-ascorbic acid served as the control group.Results:The blastocyst rate after adding 50μM L-ascorbic acid to the in vitro maturation medium was significantly higher than the control group(P<0.05),whereas 100μM L-ascorbic acid exhibited a negative effect on the blastocyst rate.The blastocyst rates for embryos cultured in 50μM L-ascorbic acid in the in vitro culture medium alone and both in vitro maturation and in vitro culture media were significantly higher than their corresponding control groups(P<0.05).The relative mRNA abundance of BAX significantly decreased in blastocysts produced after the addition of 50μM L-ascorbic acid as compared with the control group(P<0.05),whereas,for MCL1,it significantly decreased in blastocysts produced after the addition of 100μM L-ascorbic acid(P<0.05).Conclusions:The supplementation of 50μM L-ascorbic acid to in vitro maturation and in vitro culture media supports in vitro embryonic development in buffaloes by improving developmental competence and altering the expression of apoptosis-related genes.

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.

Immature Embryo Rescue and in Vitro Development Evaluation of Intraspecific Hybrids from Brazilian Seedless Grapevine“Superior×Thompson”Clones

The fruit production for export is an economically significant activity in the Valley of S?o Francisco River, especially in the irrigated lands of Petrolina-PE/Juazeiro-BA, Brazil. The development of new genetic material most suitable to the tropical climate and the demands of the consumer market have led to the selection of new seedless grapes cultivars. In this case, the use of the embryo rescue technique has produced satisfactory results for obtaining such materials, especially in the semiarid region. This study aimed to evaluate the in vitro development of intraspecific hybrids of grapevine (Vitis vinifera L.), derived from the rescue of immature embryos resultant from the crossing of “Superior Seedless” and “Thompson Seedless” Brazilian clones. To establish and develop the cultivation, the culture media was supplemented with 30 g/L sucrose, 0.1 g/L myo-inositol, 0.002 g/L glycine, 0.1 mg/L indoleacetic acid (IAA), 6.5 g/L of agar, adjusted pH to 5.7. The experiment was evaluated after 90 days. The variables measured were: number of nodes, number of leaves, plant height (cm), number of roots and length (cm) of the root system and internodes. The period of 60 days of in vitro culture of ovules resulted in the highest values of embryos (about 50%), as well as better characterized developmental stages with higher germination (47.3%). The three types of hybrid grapes evaluated in micropropagation showed very similar values of the measured parameters, even having originated from embryos of different developmental stages.

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.

Assessment of the Quality of Frozen-Thawed Semen or Epididymal Sperm in Three Native Vietnamese Pig Breeds

Our aim was to evaluate the quality of ejaculated and epididymal frozen-thawed pig sperm of endangered Vietnam native pig breeds. Ejaculated sperm was collected from live boars and epididymal sperm was collected from slaughtered boars of the MuongTe, Kieng Sat and Co BinhThuan breeds and frozen in 0.25 ml straws using a protocol established earlier for modern pig breeds. We evaluated the sperm quality after thawing in terms of motility and rates of viable and abnormal spermatozoa. Our results revealed that the sperm motility and rates of viable and abnormal frozen-thawed sperm were >30%, >44%, and <14%, respectively. The origin of sperm had an effect on the production of pig embryos in vitro. In the Co BinhThuan breed, ejaculated sperm generated higher cleavage, blastocyst and hatching rates than did the epididymal sperm (60.11% vs 56.02%, 17.23% vs 14.31%, 3.78% vs 2.34%, respectively, P < 0.05). Although no difference in cleavage rate, blastocyst formation rate and the average number of cells/blastocysts, the hatching blastocyst rate was different between the breeds (P > 0.05). In the Co BinhThuan breed, the rate of pregnancy of ejaculated groups was similar to that of the epididymal group. In conclusion, the ejaculated and epididymal sperm of native Vietnamese pigs were successfully frozen. We succeeded in creating embryos in vitro and pregnant pigs after artificial insemination from frozen-thawed semen in three native Vietnamese pig breeds for the first time. The use of the ejaculated sperm improved the production of native pig embryos in vitro efficiency.

Professor Qi Cong’ s therapeutic strategy of traditional Chinese medicine combined with different stages of assisted reproduction technology

The success rate of in vitro fertilization and embryo transfer is only about 28.5%.The combination of traditional Chinese medicine and assisted reproduction technology can effectively improve the pregnancy rate,which has good application prospect.However,there is no relevant record in traditional Chinese medicine.Unified diagnosis and treatment system has yet been established.According to many years of clinical experience,Professor Qi Cong summarized the treatment strategy of traditional Chinese medicine.Treatment of invigorating the spleen to nourish blood,reinforcing the spleen and kidney,tonifying the kidney and activating blood circulation,enriching the kidney to prevent miscarriage,cooling blood to stop bleeding are given based on the characteristics of each stage and patients'symptoms during the transplantation.Disease differentiation matters during intermittent stage.Besides,she advocated treating husband and wife together.All of the above are characterized by Chinese medicine concept of pre cultivating the loss,treating the same disease with different methods,and three categories of etiologic factors.

Propagation Techniques of Ewes at Early Age

The integration of hormone induction of young ewes,live oocyte collection,in-vitro fertilization and embryo transfer( JIVET),as a major breakthrough in livestock breeding technology,not only solves the problem of low oocytes obtained from adult ewes by hormone induction in production practice,but also significantly improves production efficiency. This technology can reduce the generation interval of cattle and sheep and speed up the acquisition of genes in genetic breeding. The application of this technology can help accelerate the cultivation process,shorten the generation interval,promote the industrialization and improve the economic benefits of beef cattle breed. This paper provides a reference for the research and application of new breeding and propagation techniques of beef cattle.

Analysis of factors related to early miscarriage after in vitro fertilization embryo transfer

Aims:This research aims to explore the factors related to early spontaneous miscarriage after in vitro fertilization and embryo transfer(IVF-ET)pregnancy,and to provide guidance for improving pregnancy outcomes.Methods:We retrospectively analyzed the data for clinical pregnant women(2591 cycles)undergoing IVF-ET at the reproductive center of Peking University People’s Hospital from January 2017 to December 2018;This included 544 ET cycles and 2047 frozen embryo transfer cycles.The analysis of factors associated with early miscarriage in the overall population of IVF/intracytoplasmic sperm injection(ICSI)pregnancies(including fresh and thawing cycles)was performed.Results:The risk of early miscarriage in the 35–39 age group was 1.35 times higher than that in the<35 age group(OR=1.35[1.05,1.73],p=0.02).In addition,the risk of early miscarriage was 3.88 times higher in the group40 years old than in the group<35 years old(OR=3.88[2.68,5.62],p<0.001).Endometrial thickness also affected the miscarriage rate;the early miscarriage risk with endometrial thickness8.5 mm was 0.78 times than that of the<8.5 mm group(OR=0.78[0.62,0.98],p=0.03).The early miscarriage rate during frozen embryo transfer was 1.48 times higher than that during fresh embryo transfer(OR=1.48[1.08,2.02],p=0.01),while in the fresh cycle,the risk of early miscarriage with high-quality embryos was 0.5 times lower than that with nonhigh quality embryos(OR=0.5[0.27,0.9],p=0.02).In the frozen cycle,the risk of early miscarriage in natural cycle transplantation was 0.73 times that in hormone replacement treatment(HRT)cycle transplantation(OR=0.73[0.54,0.97],p=0.03).Conclusions:Advanced age is an independent risk factor for early miscarriage,while endometrial thickness at the date of transplantation is an independent protective factor.The risk of early miscarriage in fresh-cycle transplanted embryos is significantly lower than that in frozen embryos,and the number of high-quality embryos in the fresh cycle lowers the miscarriage rate significantly.Natural cycle transplantation has a lower rate of early miscarriage than hormone replacement therapy.

Case of acupuncture combined with Chinese herbal medication in assistance of pregnancy after 11 failures of IVF-ET at advanced age

This paper reports one case of successful pregnancy in the assistance of acupuncture therapy for regulating menstruation and promoting pregnancy and the periodic intervention with Chinese herbal medicine in a 41-year-old patient with infertility.The patient suffered from 11 failures in the assisted reproductive technique during 3 years.This case suggests that the combined treatment with acupuncture for regulating menstruation and promoting pregnancy and the periodic intervention with Chinese herbal medicine increase the levels of follicle-stimulating hormone(FSH)and estradiol(E2),improve the oocyte quality and the uterine environment.This therapeutic method plays a positive auxiliary role in the whole process of in vitro fertilization and embryo transfer(IVF-ET)and it can be adopted as an intervention for patients with multiple assisted reproductive failures.

Pregnancy outcomes after fertility preservation in women with endometrial carcinoma and atypical endometrial hyperplasia:A systematic review and meta-analysis

Objective:The objective of the study was to compare the effects of assisted reproductive technology(ART)and spontaneous pregnancy on pregnancy outcomes in women with endometrial carcinoma(EC)and atypical endometrial hyperplasia(AEH)following fertility-sparing treatments.Methods:Relevant studies published through July 2020 were identified from PubMed and Web of Science literature searches.The pregnancy outcomes of ART and spontaneous pregnancy were summarized and compared for women with complete remission of EC/AEH after fertility-sparing treatments.A subgroup analysis was performed based on whether patients had received in vitro fertilization and embryo transfer(IVF-ET)treatment.The complete remission and recurrence rates of EC/AEH following fertility-sparing treatments were estimated.The effect of pregnancy on recurrence rates of EC/AEH was also calculated.Results:Sixteen observational studies reporting pregnancy outcomes or recurrence with ART or spontaneous pregnancy for women with EC/AEH after fertility-sparing treatments were included.The complete remission rate of EC/AEH was 81.5%(95%CI,78%–85%).Compared with spontaneous pregnancy,the pregnancy rate of ART was significantly higher(66.8%vs.43.7%,OR=2.64,95%CI 1.71–4.05,P<0.00001,I^(2)=14%).Subgroup analysis showed that the pregnancy rate of IVF-ET was significantly higher than that of spontaneous pregnancy(62.7%vs.35.1%,OR=2.85,95%CI 1.44-5.63,P=0.003,I^(2)=29%).The live birth rate of ART was significantly higher than that of spontaneous pregnancy(75.3%vs.47.8%,OR=3.96,95%CI1.76-8.77,P=0.0009,I^(2)=45%).The recurrence rate of EC/AEH was 31%(95%CI 24%–39%).Clinical pregnancy could reduce the recurrence rate of EC/AEH,but there was no significant evidence of an association.Conclusions:ART,especially IVF-ET,could significantly improve pregnancy outcomes in women with EC/AEH receiving fertility-sparing treatments.Following complete remission,ART treatment might be a better option for women with EC/AEH than spontaneous pregnancy.

Effects of Transcutaneous Electrical Acupoint Stimulation on Ovarian Responses and Pregnancy Outcomes in Patients Undergoing IVF-ET:A Randomized Controlled Trial

Objective:To evaluate the influence of different transcutaneous electrical acupoint stimulation(TEAS)modes on ovarian responses and pregnancy outcomes in patients with infertility undergoing in vitro fertilization and embryo transfer(IVF-ET).Methods:Two hundred infertility patients undergoing IVF-ET were divided randomly into experimental groups(TEAS groups:E-Ⅰ,E-Ⅱ,E-Ⅲ,and E-Ⅳ,40 cases each group)and a control group(mock TEAS group,40 patients)using the random number method.The patients in the experimental groups received TEAS treatment of 20,30,40 and 50 m A for the E-Ⅰ,E-Ⅱ,E-Ⅲand E-Ⅳgroups,respectively.The control group received a treatment of 5 m A.TEAS was applied at acupoints of Guanyuan(RN 4),Zhongji(RN 3),Sanyinjiao(SP 6),Zigong(EX-CA 1),and Taixi(KI 13),once a day for 30 min each time for a treatment period of 10-13 d.Treatment effect was assessed using the following indicators:endometrial thickness on the 6 th day of gonadotropin treatment(GN6 day),endometrial thickness on the day on chorionic gonadotropin administration(HCG day),number of ovarian follicles on HCG day,number of ova captured,amount of estrogen required for each harvested ova,number of mature ova divided by the total number of ova,percentage of high-quality embryos,and clinical pregnancy.Results:Endometrial thickness in the experimental groups on the HCG day was significantly better than that of the control group after TEAS stimulation(P=0.01).TEAS exhibited a greater impact on the number of ova captured(P=0.003).However,the effect of TEAS stimulation on the high-quality embryo rate and clinical pregnancy in patients was not statistically significant(P>0.05).Conclusions:TEAS is an effective method in improving the ovarian state.When the stimulus intensity was at 40 m A and above,it could be helpful to improve the patient’s endometrial condition and endometrial receptivity and to retrieve more oocytes.(Trial registration No.Chi CTR-TRC-11001780)

Establishing the lower limits of total serum testosterone among Chinese proven fertile men who received treatment of assisted reproductive technology

Testosterone(T)plays a crucial role in spermatogenesis because extremely low levels of intratesticular T lead to correspondingly low serum levels of total T(tT),severe disorders of spermatogenesis,and male sterility.However,there is little consensus on the lower limits of serum tT in proven fertile men undergoing assisted reproductive technology treatments in Chinese or other Asian populations.We aimed to establish the reference range of serum tT based on a population of 868 fertile Chinese men undergoing in vitro fertilization or intracytoplasmic sperm injection and embryo transfer(IVF/ICSI-ET)treatments.We defined a fertile man as having had a live baby with his partner as recorded in our IVF registration system.The lower limits of serum tT were established using a Siemens IMMULITE 2000 chemiluminescent system.The 1st,2.5th,and 5th percentiles and their 95%confidence intervals(CIs)were 3.6(95%CI:2.7–4.1)nmol l−1,4.3(95%CI:4.1–5.0)nmol l−1,and 5.6(95%CI:4.8–5.8)nmol l−1,respectively.Using the linear correlation of serum tT between the Siemens platform and a liquid chromatography–tandem mass spectrometry platform,the calculated lower limits of serum tT were also established for fertile Chinese men undergoing IVF/ICSI-ET treatments,which will benefit the clinical diagnosis and treatment of male infertility during such procedures.