Effects of repeated superovulation on developmental potential of oocytes in mice and humans / 中国组织工程研究

BACKGROUND:Superovulation is a common therapy in assisted reproductive technology.In clinical practice,some patients experience repeated superovulation to get pregnant. OBJECTIVE:To explore the effect of repeated superovulation on the developmental potential of oocytes in mice and humans. METHODS:Both animal experiments and retrospective clinical research were conducted.The animal study involved 90 SPF grade ICR 8-week-old female mice,who were randomly divided into three groups for 1,3,and 5 superovulations,respectively.The clinical study involved 306 patients who had undergone three consecutive in vitro fertilization cycles.The number of ovules obtained and embryonic development in different cycles were compared. RESULTS AND CONCLUSION:(1)The animal study indicated that repeated superovulation did not affect the embryonic development or developmental speed of mouse embryos.Similarly,there was no significant difference in the mouse blastocyst apoptosis,DNA damage,or the formation of inner cell mass and trophectoderm(P>0.05).(2)The clinical study also revealed no significant differences in the number of retrieved oocytes(8.60±5.04,8.58±4.87,and 8.38±4.63,P=0.81)and transferable embryos(2.42±1.99,2.40±1.92,and 2.64±2.00,P=0.26)over the three cycles.(3)In both the young group(<35 years)and the old group(≥35 years),the embryo quality was not affected by repeated superovulation(P>0.05).(4)These findings show that repeated superovulation does not affect the developmental potential of oocytes in mice and humans.

Impairment of superovulation on the establishment of pregnancy in mice / 解剖学报

Objective To explore whether superovulation impairs the process of pregnancy establishment in mice by changing the intrauterine environment. Methods The implantation and pregnancy of superovulated and normal mice were compared. The superovulated mice were subjected to unilateral tubal ligation on day 0. 5 and blastocysts were transplanted to the other uterine horn on day 2. 5. The number of implantation sites of bilateral uterine horn was compared. The differences between preimplantation uteri of superovulated and normal pseudopregnancy mice were compared by tissue sections and high-throughput sequencing. Bioinformatics analysis was performed on the differentially expressed genes in two groups. Results Compared with the control group, the pregnancy rate of mice in the superovulation group decreased significantly. The number of implantation sites in the superovulation group was higher than the control. There was no significant difference in the pregnancy rate of the uterine horn between the control side and the transplanted side of the superovulated mice. The endometrium was thinned and the number of glands was reduced in superovulated pseudopregnancy mice. The gene expression patterns of preimplantation uterus in superovulation pseudopregnancy and normal pseudopregnancy mice were different. There were 1097 significantly differentially expressed genes, including 752 up-regulated genes and 345 down-regulated genes. Bioinformatics analysis showed that differentially expressed genes are mainly involved in biological processes, such as decidualization, response to progesterone, positive regulation of angiogenesis. They were mainly enriched in FoxO signaling pathway, cell cycle pathway and steroid biosynthesis pathway. Conclusion Superovulation impaired the process of establishing pregnancy and altered the gene expression patterns of biomarker of uterine receptivity in mice.

The impact of BMI on the clinical outcomes of patients undergoing long-term IVF/ICSI treatment in the follicular phase / 中国医师杂志

Objective:To investigate the effect of body mass index (BMI) on the first in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) treatment for young women of childbearing age (20-35 years old) with tubal factor infertility.Methods:A retrospective analysis was conducted on the clinical data of 747 patients who underwent assisted reproductive technology (IVF/ICSI) treatment for the first time at the Reproductive Medicine Center of the First People′s Hospital of Changde City from January 1, 2019 to September 30, 2021. The patients were aged 20-35 years old and all received controlled ovarian hyperstimulation treatment using a long-term follicular protocol. Patients were divided into four groups based on their BMI values during physical examinations before assisted pregnancy treatment: the weight loss group [body mass index (BMI)<18.5 kg/m 2 in 72 cases], the normal weight group (18.5 kg/m 2≤BMI<24 kg/m 2 in 455 cases), the super recombination group (24 kg/m 2≤BMI<28 kg/m 2 in 161 cases), and the obesity group (BMI≥28 kg/m 2 in 59 cases). The differences in relevant indicators during controlled ovarian hyperstimulation treatment and pregnancy outcomes after fresh embryo transfer were analyzed among each group. Resultsl:There were no statistically significant differences in age, infertility years, primary/secondary infertility ratio, basal follicle stimulating hormone levels, and sinus follicle count among the lean, normal weight, overweight, and obese groups (all P>0.05). There were no statistically significant differences in the duration of gonadotropin (Gn) administration, the number of follicles with a diameter of ≥14 mm on the trigger day, the level of estrogen on the trigger day, the number of retrieved eggs, the number of double pronuclei, the rate of cyst formation, the rate of transplantation, and the rate of whole embryo cryopreservation among the four groups (all P>0.05). The total amount of Gn was found to be (2 374.5±892.5)U in the lean group, (2 643.8±1 213.5)U in the normal weight group, (2 950.5±1 156.5)U in the super recombinant group, and (2 949.8±1 030.5)U in the obese group, with a statistically significant difference ( P=0.001). The number of excellent embryos was (3.06±2.50) in the lean group, (2.96±2.16) in the normal weight group, (2.45±1.96) in the super recombinant group, and (2.39±2.03) in the obese group, with a statistically significant difference ( P=0.015). The embryo free rate among the four groups significantly increased with increasing BMI, and the difference was statistically significant ( P=0.039). There was no statistically significant difference in the number of embryos transferred, biochemical pregnancy rate, clinical pregnancy rate, embryo implantation rate, multiple pregnancy rate, early abortion rate, mid-term abortion rate, and live birth rate among the four groups (all P>0.05). Conclusions:In patients with simple tubal factor infertility who receive long-term follicular phase assisted fertilization treatment, an increase in BMI can significantly increase the dosage of Gn, reduce the number of excellent embryos, and increase the rate of no embryos. Although there is currently no statistical difference in various indicators of pregnancy outcomes after fresh embryo transfer among different BMI groups, there is a risk of an increase in early or mid-term miscarriage rates as BMI increases.

Effect of different ovulation induction regimens for polycystic ovary syndrome / 中国医师杂志

Objective:To explore effects of controlled ovarian stimulation (COS) protocols on pregnancy outcomes for patients with polycystic ovarian syndrome (PCOS) undergoing in vitro fertilization-embryo transfer (IVF-ET). Methods:A total of 1 032 patients with PCOS who underwent IVF-ET from September 1, 2016 to July 31, 2020 in the Reproductive Center of Hunan Provincial Maternal and Child Health Care Hospital were retrospectively analyzed. The patients were divided into modified long regimen group (group A, 126 cases), luteal phase long regimen group (group B, 185 cases), antagonist regimen group (group C, 344 cases), and progestin primed ovarian stimulation (PPOS) group(group D, 377 cases) according to different ovulation stimulation regimens. The ovulation promotion status [days of gonadotropin (Gn), total amount of Gn, estradiol (E 2) level on the day of human chorionic gonadotropin (HCG) injection, number of retrieved eggs, number of mature eggs (MII eggs), number of normal fertilized embryos (2PN), number of high-quality embryos] and the first frozen embryo transfer pregnancy status (clinical pregnancy rate, implantation rate, early abortion rate) were compared among the patients in each group. Results:(1) There was no significant difference in general clinical data between the four groups (all P>0.05). (2) The number of Gn days in group D was significantly less than that in groups A, B and C, and the total number of Gn was significantly less than that in groups A, B and C (all P<0.05); The E 2 level of patients in group C and group D on the day of hCG injection was significantly lower than that of group A and group B (all P<0.05); The number of eggs obtained and MII eggs in group C and group D were significantly lower than those in group A and group B (all P<0.05); The number of high-quality embryos and 2PN in group D were significantly different from those in group A, group B and group C (all P<0.05). (3) The clinical pregnancy rates of the first frozen embryo transfer after whole embryo cryopreservation in group A, group B, group C and group D were 54.72%(29/53), 56.79%(46/81), 52.56%(82/156) and 54.32%(195/359), respectively, with no significant difference among the four groups (all P>0.05). There was no significant difference in embryo implantation rate and early abortion rate among the four groups (all P>0.05). Conclusions:The modified long regimen, luteal phase long regimen, antagonist regimen and PPOS regimen can achieve better pregnancy outcomes in patients with PCOS. Among them, PPOS regimen can reduce the amount and time of Gn, and frozen embryo transfer does not affect the pregnancy outcome of patients. It can be used as one of the priority recommended strategies for PCOS patients who plan to undergo frozen embryo transfer.

Effectiveness, safety and cost of urinary follicle stimulating hormone in controlled ovarian stimulation in China: multi-center retrospective cohort study of 102 061 in vitro fertilization cycles / 中华妇产科杂志

Objective:To explore the effectiveness, safety and cost between urinary follicle stimulating hormone (uFSH) and recombinant follicle stimulating hormone (rFSH) in controlled ovarian stimulation (COS) in China.Methods:Data were collected from 16 reproductive centers in China covering oocytes collection time from May 1, 2015 to June 30, 2018. Eligible patients were over 18 years old, adopting COS with uFSH (uFSH group) or rFSH (rFSH group) as start gonadotropins (Gn), and using in vitro fertilization (IVF) and (or) intracytoplasmic sperm injection for fertilisation, excluding frozen embryo recovery cycle. Generalised estimating equation was used to address the violation of independency assumption between cycles due to multiple IVF cycles for one person and clustering nature of cycles carried out within one center. Controlling variables included age, body mass index, anti-Müllerian hormone level, cause of infertility, ovulation protocol, type of fertilisation, number of embryos transferred, number of days of Gn use.Results:Totally 102 061 cycles met eligibility criteria and were included in the analyses. In terms of effectiveness, after controlling relevant unbalanced baseline characteristics, compared with rFSH group, the high oocyte retrieval (>15 oocytes was considered high retrieval) rate of uFSH group significantly decreased in gonadotropin-releasing hormone agonist protocol ( OR=0.642, P<0.01) and in gonadotropin-releasing hormone antagonist protocol ( OR=0.556, P=0.001), but the clinical pregnancy rate per transfer cycle and the live birth rate per transfer cycle significantly increased ( OR=1.179, OR=1.169, both P<0.01) in both agonist and antagonist protocols. For safety, multiple analysis result demonstrated that in the agonist protocol, compared with rFSH group, the incidence of moderate to severe ovarian hyperstimulation syndrome of uFSH group significantly decreased ( OR=0.644, P=0.002). The differences in ectopic pregnancy rate and multiple pregnancy rate between the uFSH and rFSH groups were not significant ( P=0.890, P=0.470) in all patients. In terms of cost, compared with rFSH group, the uFSH group had lower total Gn costs for each patient ( P<0.01). Conclusion:For patients who underwent COS, uFSH has better safety, and economic profiles over rFSH in China.

Relationship between superovulation and embryo production with ovarian follicular population before superovulatory treatment in Brazilian Bergamasca sheep / [Relação da resposta superovulatória e produção embrionária com a população folicular ovariana antes do tratamento superovulatório de ovelhas da raça Bergamácia Brasileira]

The objective of this study was to quantify the superovulatory response and embryo production of Brazilian Bergamasca sheep and to evaluate the link to the follicular condition before superovulatory treatment, as a reference for selection of donors with potential for superovulation. Follicular population of twenty-three sheep was evaluated by ultrasound during metestrus phase of the estrous cycle and divided into groups of low, medium and high follicular population. Subsequently, they were synchronized, superovulated with 133mg of pFSH, mated and subjected to embryo collection. The superovulatory response (9.0±3.3 vs 10.7±6.2 vs 13.8±7.1) and embryo production (4.0±3.8 vs 2.6±2.0 vs 1,8±4.0) were similar between groups (P>0.05). There was a positive correlation between the number of follicles during the metestrus phase and the number of corpus luteum with premature regression (PLR) (0.52) and a negative correlation between the recovery rate and PLR (-0.44) (P<0.05). The sheep that presented PLR had more follicles during metestrus (16.9±7.8 vs 12.7±3.2) and lower embryo recovery rate (38.8±29.3 vs 72.2±29.9) than those with functional CLs (P<0.05). Follicular quantification during metestrus phases was unable to identify donors with high embryo production. Animals with PLR had a larger follicular population during metestrus and lower embryo recovery rate.(AU)

O objetivo deste trabalho foi quantificar a resposta superovulatória e a produção embrionária de ovelhas Bergamácia Brasileira e relacioná-las com a condição folicular antes do tratamento superovulatório, como referência para seleção de doadoras com potencial para superovulação. Vinte e três ovelhas foram avaliadas quanto à população folicular por ultrassonografia na fase de metaestro do ciclo estral e divididas em grupos com baixa, média e alta população folicular. Posteriormente foram sincronizadas, superovuladas com 133mg de pFSH, acasaladas e submetidas à coleta de embriões. A resposta superovulatória (9,0±3,3 vs. 10,7±6,2 vs. 13,8±7,1) e a produção embrionária (4,0±3,8 vs. 2,6±2,0 vs. 1,8±4,0) foram semelhantes entre os grupos (P>0,05). Houve correlação positiva entre o número de folículos no metaestro e o número de corpos lúteos com regressão prematura (RPCL) (0,52) e correlação negativa entre a taxa de recuperação e RPCL (-0,44) (P <0,05). As ovelhas que apresentaram RPCL tiveram mais folículos no metaestro (16,9±7,8 vs. 12,7±3,2) e menor taxa de recuperação embrionária (38,8±29,3 vs. 72,2±29,9) do que as que apresentaram CLs funcionais (P<0,05). A quantificação folicular nas fases de metaestro não foi capaz de identificar doadoras com alto potencial de produção embrionária. Animais com RPCL tiveram maior população folicular no metaestro e menor recuperação de embriões.(AU)

Comparison of intrauterine insemination and timed intercourse following controlled ovarian hyperstimulation in unexplained infertility: a randomized controlled trial

Background: Being a diagnosis of exclusion the treatment options of unexplained infertility are often empiric. There is significant dilemma regarding the superiority of one over another. Despite increasing use of intrauterine insemination (IUI) in adjunct to controlled ovarian hyperstimulation (COH) there is scarcity of randomized controlled trials (RCT) from developing countries. Objective was to compare IUI and timed intercourse (TI) in super ovulated cycles among couples with unexplained infertility over one year.Methods: In this prospective randomized controlled trial total 85 patients were randomly assigned into group 1 (COH with IUI, N= 44) and group 2 (COH with TI, N=41). Patients underwent COH using sequential Clomiphene Citrate and injection human menopausal gonadotrophin followed by IUI in group 1 and timed intercourse in group 2. Either protocol was repeated for three consecutive cycles. Finally, both groups were compared for clinical pregnancy rate, adverse effects and acceptability of the treatment process and outcome. Comparison was done by Student’s unpaired t test for continuous and 2-tailed chi square test for categorical variables.Results: Clinical pregnancy rates following COH/IUI and COH/TI were 13.64% and 19.51% respectively. There was observable difference in the acceptability of the outcome (38.64% in IUI and 56.09% in TI group). All the results including complications and side effect rates were statistically insignificant.Conclusions: Present study failed to show any improvement of pregnancy rates following addition of IUI over TI and it raised the probability that the outcome of the procedure may not be well accepted.

Changes in Biochemical Profile of Superovulated Sahiwal Cows through Hormonal Manipulation at Mid Luteal Phase of Estrous Cycle

In the present study, insulin and insulin-like growth factor-I (IGF-I) administered during mid-luteal phase of the estrous cycle and effect on biochemical profiles of Sahiwal donor cows was noted. Altogether eighteen cows (n=18) were selected and divided into 3 groups; control (n=6, untreated), T-I (n=6, Insulin-treated) and T-II (n=6, IGF-I treated). Superovulatory treatment was started on day 9th of the estrous cycle. With the 6th dose of FSH, prostaglandin was injected to induce superovulatory estrus. The superovulated cows were bred and superovulatory response of each animal was recorded. The collections of embryos were done non-surgically on the 7th day of superovulatory estrus. About 15 ml blood without anticoagulant was collected on days 5,7,9,11,13,15,17,19 and 21st or day of embryo recovery where day 0 of estrous cycle was taken as the day of estrus. Serum was separated, centrifuged at 3000 rpm for 15 minutes and transferred into sterilized serum vials. All samples were stored at -20 oC till analysis. Serum glucose, cholesterol, total protein, urea, and creatinine were estimated by Span Diagnostic Kits. The concentration of serum glucose increased significantly in insulin-treated and IGF-1 treated Sahiwal donor cows. It may be concluded that exogenous insulin and IGF-1 administration during mid-luteal phase may be helpful in follicular and embryonic development by increasing the level of serum glucose. The concentration of serum cholesterol, total protein, urea, and creatinine remains unaffected

Induction of superovulation in mature mice and rats using serum of spayed female dogs / 한국실험동물학회지

The following experiments were designed to examine the effect of serum of spayed dogs on superovulation response in mice and rats. In Experiment 1, female mice at diestrus (n=30) were divided into three equal groups and superovulated with either administration of 5 IU pregnant mare serum gonadotropin (PMSG) or recombinant follicle stimulating hormone (rFSH) (reducing dose from 2.5 to 0.5 IU) and 5 IU human chorionic gonadotropin (hCG) administered 48h later. Serum of spayed dogs was administered intraperitoneally at a reduced dose from 0.1 to 0.025 mL in a 48 h period. In Experiment 2, female rats (n=30) at diestrus stage were divided into three equal groups. Superovulation was induced using either 30 IU PMSG, or a dose reduced from 5 to 1 IU rFSH and 25 IU hCG administered 48h later. Serum of spayed dogs was administered in a reduced dose from 0.6 to 0.1 mL in a 48 hour period. Female mice and rats were mated 24 h following hCG administration. On day 14 after mating, animals were euthanized and ovarian sections were fixed for histopathological evaluation and corpus luteum (CL) counting. No significant difference observed in mean (±SEM) number of CLs between the PMSG group and the mice that received serum of spayed dog (10.4±1.3 vs 9.2±1.0). Mean (±SEM) number of CLs tended to be lower in rats that received serum of spayed dog than those of rats which received either PMSG or rFSH (15.1±1.9 vs 23.6±3.1 and 23.1±2.9, P=0.06, respectively). In conclusion, serum of spayed dogs is able to induce a superovulatory response in mice and rats.

Estudo da vascularização folicular e do corpo lúteo de éguas cíclicas tratadas com extrato de pituitária equina utilizando ultrassom Doppler colorido / Study of follicular and corpus luteum vascularization in mares treated with Equine Pituitary Extract using ultrasound color Doppler

Informações sobre a vascularização da parede folicular e do corpo lúteo equino, associadas à superovulação, são escassas. Com o objetivo de avaliar o efeito superovulatório do extrato de pituitária equina (EPE) no fluxo sanguíneo folicular e luteal, foram utilizadas seis éguas Puro Sangue Árabe, em dois ciclos estrais (controle e tratamento). As éguas foram monitoradas diariamente por ultrassonografia modo B, até que os folículos atingissem diâmetro de 23mm (desvio). No ciclo tratamento, as éguas receberam 8mg de EPE, uma vez ao dia, por via IM, até que dois ou mais folículos atingissem o diâmetro entre 32 e 35mm. A ovulação foi induzida com acetato de deslorelina, quando os folículos atingiram, no mínimo, 35mm. No momento do desvio folicular, da indução da ovulação e do último exame pré-ovulatório, foi utilizada a ultrassonografia modo B para medir o diâmetro dos folículos e, no oitavo dia pós-ovulação, para a área do corpo lúteo (CL). Utilizou-se também ultrassonografia com Doppler colorido para avaliar a perfusão sanguínea da parede folicular e do parênquima luteal. No ciclo controle, foi realizado o mesmo procedimento, exceto pelo uso do EPE. Os dados foram submetidos à análise de variância, com nível de significância de 5%. Não foi observado efeito do EPE sobre o número de ovulações, o diâmetro dos folículos, a vascularização da parede folicular e a concentração sérica de estrógeno. Os animais, tratados ou não, apresentaram CLs funcionais, não havendo diferença na área do parênquima ou da vascularização luteal, nem na concentração sérica de progesterona, no oitavo dia após a ovulação. Foi observado que o EPE proporcionou um maior número de folículos subordinados no momento da indução da ovulação do folículo dominante (P ≤ 0,05). Embora esses folículos não tenham chegado a ovular, concluiu-se que o EPE atuou no crescimento de folículos, que podem ser utilizados em outras biotécnicas, como a transferência de oócitos, com maior aproveitamento da reserva folicular de ovários equinos.(AU)

Knowledge about follicle and corpus luteum vascularization associated with superovulation in mares is scarce. Aiming to evaluate the effect of equine pituitary extract (EPE) on superovulation, the experiment was conducted using six mares Purebred Arabian in two estrous cycles (control and treatment). The mares were synchronized, and monitored daily by ultrasound B mode until the follicles reached diameter ≤ 23 mm (deviation). In the treatment cycle, from the deviation, mares received 8 mg of EPE, once a day, intramuscularly, until two or more follicles reached a diameter between 32 and 35 mm. Ovulation was induced with deslorelin acetate when follicles reached at least 35 mm. At the time of follicular deviation, induction of ovulation and final preovulatory exam, it was used B-mode ultrasound to measure the diameter of follicles and on the eighth day after ovulation to measure the area of the corpus luteum (CL); color Doppler was also used to assess blood perfusion of the follicle wall and luteal parenchyma. In the control cycle was performed the same procedure except for the use of EPE. Data were subjected to analysis of variance, with 5% significance level. There was no effect of EPE on ovulation number, diameter of follicles, vascularity of the follicular wall and serum estrogen concentration. The animals treated or not, showed functional CLs, with no difference in parenchymal area or luteal vascularization, or in serum progesterone concentration on the eighth day after ovulation. It was observed that the EPE provided a greater number of subordinate follicles at the time of induction of ovulation of the dominant follicle. Although these follicles have failed to ovulate, it was concluded that EPE influenced the follicles growth, and it can be used in other biotechnologies, with greater utilization of equine ovarian follicular reserve.(AU)

Effect of serum containing Jinghou Zengzhi Recipe on expression of GDF9 and Bim in ovarian COCs and CCs in controlled ovarian hyperstimulation rats / 重庆医学

Objective To observe the effects of serum containing Jinghou Zengzhi Recipe (JHZZ) on the expression of growth differentiation factor 9 (GDF9) and Bim in cumulus-oocyte complexes (COCs) and cumulus cells (CCs) of controlled ovarian hyperstimulation (COH) rats,and to investigate its curative effect and mechanism.Methods The rat COH ovarian model was prepared for collecting COCs and CCs,which were respectively co-cultured in vitro with the blank serum of female COH SD rat and serum containing JHZZ,and the each group was divided into 3 subgroups:the blank serum group (blank group),serum containing JHZZ group (control group) and serum containing JHZZ plus GDF9 receptor blocker group (experimental group),total 6 subgroups.COCs and CCs were collected after 24 h.The expression levels of GDF9 and Bim mRNA were detected by real-time quantitative PCR.The GDF9 protein expression levels were detected by Western blot.Results (1) The expression level of GDF9 mRNA in control group COCs was obviously higher than that in the control group and experiment group COCs (P＜0.05);the Bim mRNA expression level in COCs of control group and experiment group was significantly lower than that in COCs of blank group (P＜ 0.05);the GDF9 protein expression level in the control group COCs was significantly higher than that in the blank group COCs (P＜0.05),and also higher than that in the experiment group COCs,but the difference was not statistically significant (P＞0.05).(2)The comparison results of GDF9 mRNA expression level in CCs among 3 groups were consistent with those in COCs;the Bim mRNA expression level in CCs of control group and experimental group was significantly lower than that in the blank group (P＜ 0.05);the Bim mRNA expression level in the control group CCs was significantly lower than that in the experimental group CCs (P＜0.05);the GDF9 protein expression level in the control group CCs was significantly higher than that in the blank group and experimental group CCs (P＜0.05).(3)The GDF9 mRNA expression level in the control group COCs was significantly higher than that in CCs (P＜0.05),and the other inter-group comparisons between COSs and CCs had no statistical difference (P＞0.05).Conclusion Serum containing JHZZ can increase the GDF9 expression level in COCs and CCs,maintain the low expression of Bim in COCs and CCs,inhibit the ovarian cells apoptosis and promote the follicular development;COCs is more conducive to the expression of GDF9 mRNA compared with CCs eliminating oocyte.

Repeated superovulation induction after failure in previous IVF-ET cycles with the ultra-long protocol: Analysis of outcomes of clinical pregnancy / 中华男科学杂志

Objective@#To analyze the clinical outcomes of repeated superovulation induction in patients with adenomyosis or moderate to severe pelvic endometriosis after failure in previous IVF-ET cycles with the ultra-long protocol.@*METHODS@#We retrospectively analyzed the clinical data about 37 patients with adenomyosis or moderate to severe pelvic endometriosis in our center from 2009 to 2013, who underwent repeated IVF-ET after failure in the previous cycles with the ultra-long protocol, namely by injection of 2－6 ampoules of 3.75 mg gonadotropin-releasing hormone agonist (GnRH-a). All the patients met the following requirements: hCG-negative at 14 days after transfer, within 3－7 days after menstruation, and properly down-regulated serum follicle stimulating hormone (FSH) (<10 mIU/ml), luteinizing hormone (LH) (<10 mIU/ml), estradiol (E2) (<30 pg/ml), follicle diameter (<10 mm) and endometrial thickness, and received GnRH (Gonal-F, Serono) for ovulation induction. We compared the clinical and laboratory data and pregnancy outcomes between the first and repeated cycles before and after ovulation induction.@*RESULTS@#The repeated cycles, as compared with previous ones, showed significant increases in the antral follicle count (AFC) on the first day of stimulation (7.55 ± 1.86 vs 6.45 ± 2.5, P<0.05), number of follicles =≥14 mm in diameter on the hCG trigger day (7.81 ± 3.6 vs 5.56 ± 3.68, P<0.05), level of E2 (［2 362.15 ± 1 210.49］ vs ［1 749.22 ± 1 139.44］ pg/ml, P<0.05), and numbers of oocytes retrieved (7.51 ± 3.23 vs 4.78 ± 3.41, P<0.05) and embryos transferred (2.00 ± 0.33 vs 1.50 ± 0.67, P<0.05), exhibited a remarkably reduction in the dose of GnRH (［1 791.65 ± 1 889.41］ vs ［3 439.56 ± 1 836.53］ IU, P<0.05), and achieved a clinical pregnancy rate of 62.16%.@*CONCLUSIONS@#With proper reduction of the FSH, LH and E2 levels and follicle diameter, repeated superovulation induction for IVF-ET can improve the ovarian response and pregnancy outcomes of the patients with adenomyosis or moderate to severe pelvic endometriosis after failure in the previous IVF-ET cycles with the ultra-long protocol.

Super pregnancy in a BALB/c mouse superovulated with PMSG / 한국실험동물학회지

This paper reports a case of super pregnancy in a BALB/c mouse pregnant with 30 pups following induction of superovulation using a PMSG-hCG protocol. Superovulation was induced in 10 mice by injecting 5 IU PMSG followed by 5 IU hCG 48 hours later. Immediately after injection of hCG, animals were placed with males at a ratio of 1 to 1 for 24 hours. On day 14 after mating, animals were killed by cervical dislocation and the uterus was examined for pregnancy and the number of fetuses. The mean (±SEM) number of fetuses observed in ten mice was 5.4±3.18 with an unexpectedly super pregnant mouse bearing 30 fetuses on day 14 of pregnancy.

Produção de embriões em ovinos Morada Nova e Somalis Brasileira / Embryo production in Morada Nova and Somalis Brasileira ewes

Superovulatory response and embryo yield in 19 Morada Nova and 20 Somalis Brasileira ewes was analyzed. All animals were synchronized with the insertion of an intravaginal device (CIDR®) on Day 0, replaced by a new device on Day 7, which remained in place until Day 14 and superovulated with 133mg of porcine FSH (pFSH) in decreasing doses at 12h intervals from Day 12 until Day 15 of the treatment, and a single dose of equine chorionic gonadotropin (eCG, 200UI) on Day 14 (i.e., administered in CIDR removal). Fifty hours after CIDR® removal, females were inseminated by laparoscopy. All embryos were recovered by laparotomy 5 days after insemination. Sheep which responded to the superovulation protocol (P>0.05) included 74% of the Morada Nova ewes and 50% of the Somalis Brasileira ewes. Morada Nova showed better results (P<0.05) than Somalis Brasileira in number of ovulations (15.38 ± 5.24 vs. 10.56 ± 2.83), total structures (11.00 ± 7.55 vs. 3.33 ± 1.94) and embryo yields (6.79 ± 5.35 vs. 2.90 ± 2.18). Despite the high fertilization rate, degenerate embryo rate was high for both breeds, with an overall rate of 39% (57/145). In conclusion, superovulatory response and embryo yields in Morada Nova ewes were considered sufficient to justify the use of this procedure in genetic resources conservation programs. However, improvements to embryo quality and control of precocious regression of corpus luteum are necessary to produce better results in the MOET program, with minimal variations and maximum embryo yield in Morada Nova and Somalis Brasileira ewes.(AU)

Pretreatment of normal responders in fresh in vitro fertilization cycles: A comparison of transdermal estradiol and oral contraceptive pills / 대한생식의학회지

OBJECTIVE: The aim of this study was to investigate the impact of pretreatment with transdermal estradiol (E₂) compared to oral contraceptive pills (OCPs) on controlled ovarian stimulation (COS) response in normal responders undergoing fresh in vitro fertilization (IVF)-embryo transfer (ET) cycles. METHODS: A retrospective cohort study was performed of normal responders undergoing fresh IVF-ET cycles who received pretreatment with transdermal E₂ versus OCPs prior to fresh IVF-ET. The total days of ovarian stimulation, total dosage of gonadotropins, total number of oocytes, and mature oocytes retrieved were noted. Pregnancy outcomes after ET were also recorded. RESULTS: A total of 2,092 patients met the inclusion criteria: 1,057 and 1,035 patients in the transdermal E₂ and OCP groups, respectively. Patients in the OCP group had a longer duration of COS (10.7±1.63 days, p<0.01) than the E₂ group (9.92±1.94 days). Patients in the OCP group also required higher cumulative doses of gonadotropins (2,657.3±1,187.9 IU) than those in the E₂ group (2,550.1±1,270.2 IU, p=0.002). No statistically significant differences were found in the total and mature oocytes retrieved or in the rates of biochemical pregnancy, clinical pregnancy, spontaneous miscarriage, and live birth between the groups. CONCLUSION: Our findings suggest that compared to OCPs, pretreatment with transdermal E₂ is associated with a shorter duration of ovarian stimulation and lower gonadotropin utilization, without compromising the oocyte yield or pregnancy outcomes in normal-responder patients undergoing fresh IVF.

Effects of IL-6 on the development of zygote in mice after controlled ovarian hyperstimulation / 解放军医学杂志

ObjectiveTo study the effect of IL-6 on the development of zygotes of mice after controlled ovarian hyperstimulation. Methods The present experiment included three parts: a) Addition of IL-6: 80 female ICR mice were divided into 7 groups by random number table including 6 groups of superovulation (10 each) and a group of natural ovulation cycle (n=20). According to addition of IL-6 in different concentration to culture media, the superovulated ICR mice were divided into superovulation control group (0pg/ml IL-6 group), 1pg/ml IL-6 group, 5pg/ml IL-6 group, 10pg/ml IL-6 group, 25pg/ml IL-6 group, and 50pg/ml IL-6 group, with ICR mice in natural ovulation cycle served as control. b) Addition of IL-6 receptor antibody (RA): 90 female ICR mice were divided into 7 groups according to the random number table, including 5 groups of superovulation (10 each) on the basis of addition of different concentrations IL-6 and IL-6 RA to culture media (0pg/ml IL-6+RA groups, 1pg/ml IL-6+RA group, 5pg/ml IL-6+RA group, 10pg/ml IL-6+RA group, 25pg/ml IL-6+RA group), and 2 groups of normal natural cycle (20 each), including control group and the control group+IL-6 RA (100pg/ml) group. Mice in normal control group conceived naturally while those in superovulation group conceived after superovulation. The zygotes were collected and cultured in vitro for 1 day till the formation of 2-cell embryos, then the rate of 2-cell formation was observed under microscope. Experiments of each group were repeated three times. c) Immunofluorescence identification: 10 female ICR mice were divided into control group and superovulation group (5 each) by random number table method. The expressions of IL-6 in zygotes were determined with confocal immunofluorescence method. Results IL-6 addition experiment: the rate of 2-cell formation was significantly lower (P<0.05) in superovulated control group, 1pg/ml IL-6, 25pg/ml IL-6 and 50pg/ml IL-6 groups than in control group (P=0.023, P=0.026, P=0.000 and P=0.000, respectively). IL-6 receptor antibody (RA) addition experiment: compared with normal control group, the rate of 2-cell formation decreased in superovulation group (P=0.017), so did in other IL-6 RA groups (P=0.000). Immunofluorescence identification: the expression of IL-6 in zygote was obviously lower in superovulation group than in control group. Conclusion Controlled superovulation can reduce the expression of IL-6 in zygote, and it may be related to its effect on embryonic development of mice.

The optimization scheme of Mongolian gerbils superovulation induced by PMSG and hCG / 中国比较医学杂志

Objective To optimize the scheme of Mongolian gerbil superovulation.Methods Based on the analysis of the animal-week-old, the dose and time interval of hormone, we got the best animal-week-old, dose and time interval of hormone for Mongolian gerbil superovulation.Results The 6 week old female Mongolian gerbils which injected of 10 IU PMSG and followed by 10 IU hCG in 70 hours later could get the best superovulation.We collected eggs at 16 hour after mate with male gerbils.The ovum pick-up rate reached 80%, the number of oocytes were 32.6 ±3.0, the number of the fertilized egg developed to 2-cell are 24.8 ±5.4.Conclusions This study summarizes the optimization scheme of Mongolian gerbil superovulation induced by PMSG and hCG and it supported the foundation for Mongolian gerbils embryo biotechnology.

Redução da resposta superovulatória de fêmeas bovinas superestimuladas com FSH em doses split / Reduced superovulatory response of bovine females overstimulated with FSH in split doses

Objetivou-se avaliar a superovulação (SOV) de vacas zebuínas, utilizando protocolo convencional ou protocolo com número menor de aplicações e similar dosagem (dose split). Utilizaram-se 16 fêmeas (total 32 SOV), com idade entre 17-42 meses e escore de condição corporal 2,5-4 (escala de 1-5), em delineamento tipo cross-over. No início do tratamento (D0), os animais receberam um dispositivo de progesterona e 2mg de benzoato de estradiol. As fêmeas do grupo convencional receberam 250UI de FSH/LH divididas em oito doses decrescentes administradas em intervalos de 12h (FSH/LH no D4, D5, D6 e D7 no período da manhã e tarde, nas respectivas dosagens: 50,0 UI; 37,5 UI; 25,0 UI; 12,5 UI). No D7 pela manhã, as fêmeas foram tratadas com 150μg de D+cloprostenol, e a remoção da progesterona foi realizada no D7 à tarde. As fêmeas do grupo split também receberam 250 UI de FSH/LH. No D4 de manhã, administraram-se 62,5 UI de FSH/LH via IM e 125 UI por via SC. Quarenta e oito horas após (D6) administraram-se 62,5 UI via SC e na manhã do D7 foi removida a progesterona e aplicaram-se 150μg de D+cloprostenol. As fêmeas de ambos os grupos receberam 50μg de análogo de GnRH no D8 pela manhã e foram inseminadas 12 e 24 horas após. No D15 realizou-se a colheita dos embriões em ambos os tratamentos. Avaliou-se a resposta superovulatória pela contagem do número de folículos e CLs de cada ovário, com auxílio de ultrassom. Todas as variáveis foram submetidas ao teste T de Student para amostras pareadas. Houve diferença (P<0,05) na quantidade de folículos acima de 8mm no D8 (9,06±4,54 e 5,50±4,59); número de CLs no dia da colheita (8,12± 3,26 e 4,69±3,46), número de embriões totais (6,69±3,05 e 3,37±2,50) e de embriões viáveis (5,25±2,29 e 2,37±1,78) nas vacas do grupo convencional em relação às do split, respectivamente. Conclui-se que o protocolo split tem pior resposta superovulatória e de produção in vivo de embriões, em vacas zebuínas, quando comparado ao protocolo convencional...

The aim of this study was to evaluate the superovulation (SOV) response of zebu cows, using conventional protocol or other protocol with similar dose but smaller number of applications (split dose). 16 females (32 SOV), aged 17-42 months, and body condition score of 2.5-4 (1-5 scale) were used in randomized cross-over. At the start of treatment (D0), the animals received progesterone device and 2mg of estradiol benzoate. The females from the conventional group received 250 IU of FSH/LH divided into eight decreasing doses administered at intervals of 12 hours (FSH/LH in D4, D5, D6 and D7, with their respective strengths: 50.0 IU, 37.5 IU, 25.0 IU, 12.5 IU). In D7, the females were treated with 150μg of D+cloprostenol, and the removal of progesterone device was held in the afternoon. The females from the split group also received 250 IU of FSH/LH. In the morning D4 was administered in 62.5 IU FSH/LH IM and 125 IU subcutaneously. Forty eight hours later (D6) 62.5 IU was administered subcutaneously in the morning and on D7, the progesterone device was removed and 150 μg of D+cloprostenol was applied. The females in both groups were given 50 μg of GnRH in the morning and on D8 were inseminated after 12 and 24 hours. On D15 the embryo collection was performed in both treatments. The evaluation of superovulatory response was done by counting the number of follicles and corpus luteum (CL) in each ovary, with the aid of ultrasound. The variables were assessed by Student's t test for paired samples...

Use of optimized superovulation conditions for C57 BL/6 J mouse purification / 中国比较医学杂志

Objective To define the best dose of PMSG/hCG for superovulation and optimize the biological purification system for genetic-modified mice of C57BL/6J background, we injected different dose of PMSG/hCG to promote superovulation, and carried out in vitro fertilization and embryo transplantation.Methods 5-week old female C57BL/6J mice were injected with 4 different doses(5 IU,7.5 IU,10 IU and 15 IU, respectively) of PMSG/hCG.Then the mice were mated with wild type male mice and embryos were obtained by cesarean section.In another approach, oocytes of superovulated mice were fertilized in vitro and then transplanted into surrogate female mice . Results In the cesarean section group, the highest rate of pregnancy was 89.00 ±19.05% in the 5 IU injection group.But the difference compared with other groups was not statistically significant.In the embryo transplantation group, the average number of oocytes from 10 IU injection group was 30.33 ±0.89, and the average number of 2-cell embryo was 23.78 ±0.19, significantly higher than those in the other three groups.Conclusions Administration of 5 IU PMSG/hCG will improve mating efficiency in cesarean section-based purification of C57BL/6J mice.10 IU PMSG/hCG is the best dose for superovulation of C57BL/6J mice.The number of 2-cell embryos is maximized when 10 IU PMSG/hCG is used in the embryo transplantation.

Effect of superovulation on uterine and serum biochemical parameters and its potential association with transferable embryos in Holstein dairy cows

The objective of this study was to determine the effects of superovulation (SOV) on serum and uterine biochemical parameters, uterine bacteriology and cytology and number of transferable embryos (TE). Dairy cows were placed on a Presynch/CIDR Synch protocol. The SOV group was superovulated, induced in estrus, and inseminated, whereas the control group was induced in estrus and inseminated without SOV. Uterine bacteriology and cytology and uterine and serum biochemical parameters were measured at day 7 of the estrous cycle to start the SOV protocol, as well as on the day of embryo recovery (DER). The SOV group produced 7.5 +/- 6.7 oocytes/embryos, of which 3.4 +/- 4.7 were TE. Serum urea and E2 and uterine Glu, CK, LDH, TP, P4 and PGFM in the control group and serum P4 and PGFM and uterine LDH and PGFM in the SOV group were significantly higher (p < 0.01) at DER than day 7. At DER, uterine urea, LDH, PGFM and TP and serum urea, LDH, PGFM, and P4 concentrations were higher (p < 0.01) in the SOV group than the control. There was no significant variation in uterine bacteriology or cytology. Overall, these results infer that SOV affects both serum profile and uterine secretions, and that these changes may influence the number of TE.