hCG Injection Research Articles

Study on the Influencing Mechanism of Human Chorionic Gonadotropin (hCG) on Oocyte Maturation in Patients with Polycystic Ovary Syndrome.

The study was aimed at investigating the influence of human chorionic gonadotropin (hCG) hormone on oocyte maturation in the patients with polycystic ovary syndrome (PCOS). A total of 54 patients with PCOS who received in vitro maturation (IVM) treatment in the Cheeloo College of Medicine, Shandong University, were divided into two groups: one group who underwent hCG injections was the observation group (OG; n = 27) and other was the control group (CG; n = 27) with no hCG injection. The oocyte development and the expression of steroid hormone synthesis-related genes including gonadotropin-releasing hormone receptor (GnRHR), Conexin43, epidermal growth factor-related genes, luteinizing hormone/choriogonadotropin receptor (LHCGR), epiregulin (EREG), and vascular endothelial growth factor (VEGF) were examined. The human ovarian granulosa cell line (SVOG cells) and ovarian epithelial cell line (HOSEpiC cells) were employed to analyze the effect of hCG on the biological behaviour of cells. As a result, OG showed higher normal fertilization, cleavage, and high-qualified embryo rate than CG. Expression levels of GnRHR, Cx43, LHCGR, EREG, and VEGF were significantly elevated in granulosa cells in the OG group. Western blot revealed that phosphoinositide 3-kinase (PI3K), protein kinase B (AKT), and rapamycin (mTOR) proteins were decreased in granulosa cells under hCG intervention. A biological behaviour test indicated that the multiplication capacity of hCG-intervened SVOG and HOSEpiC was increased, while the apoptosis was decreased. In conclusion, hCG could accelerate follicular development and oocyte maturation by activating oocyte maturation genes in PCOS patients, which could significantly improve and popularize the application of IVM technology.

Dual trigger with gonadotropin releasing hormone agonist and human chorionic gonadotropin improves live birth rates in POSEIDON group 3 and 4 expected poor responders

Objective Aim of this study was to evaluate the effects of dual triggering with gonadotropin releasing hormone agonist and human chorionic gonadotropin (hCG) on outcomes of fresh embryo transfers following gonadotropin antagonist cycles in POSEIDON group 3 and group 4 low prognosis women and to compare the outcomes with hCG-only triggering. Methods This study was conducted by retrospective analysis of patients with expected poor ovarian response (POSEIDON group 3 and 4) that underwent fresh embryo transfers following in-vitro fertilization/intracytoplasmic sperm injection cycles with either dual triggering or hCG-only triggering between January 2010 and April 2020. A total of 1068 women that underwent dual triggering and 1931 that underwent hCG-only triggering were included in the study. Results Number of retrieved oocytes, M2 oocytes, oocyte maturation rate, fertilization rate, obtained 2PN embryos, implantation rate, clinical pregnancy rate and live birth delivery rates were found significantly higher in dual-triggering group in comparison to hCG-only group (p < 0.001, p < 0.001, p < 0.001, p < 0.001, p < 0.001, p < 0.001, p = 0.02, p < 0.001 respectively). Conclusion Dual trigger with concomitant injections of GnRH agonist and hCG in GnRH antagonist cycles appears to improve IVF outcomes, increase quality of embryos, reduce miscarriage rates and consequently increase live birth delivery rates in POSEIDON groups 3/4 poor responders.

Intrauterine Instillation of Human Chorionic Gonadotropin with Intrauterine Insemination Catheter Around the Golden Time of Embryo Transfer Does Not Improve In Vitro Fertilization /Intracytoplasmic Sperm Injection Outcomes in Infertile Women: A Randomized Controlled Trial.

We set out to explore the effect of intrauterine human chorionic gonadotropin (hCG) instillation by intrauterine insemination (IUI) catheter before embryo transfer (ET) on assisted reproductive technologies (ART) outcomes of infertile women. One hundred women with infertility who were scheduled for in vitro fertilization (IVF)/intracytoplasmic sperm injection (ICSI) cycles were included in the study. They were randomly devoted to two groups: experimental (n= 50) and control (n= 50). In the experimental group, 500 IU hCG passed into the internal cervical orifice via IUI catheter within 15 minutes before the transfer of fresh or vitrified cleavage-stage embryos. The control group underwent the same ET procedure without prior injection of hCG. None of the outcomes showed a statistically significant difference between the two groups. In the intervention and control groups, respectively, biochemical pregnancies rates were 26% and 18%, implantation rates were 13.5% and 8.6%, clinical pregnancies rates were 22% and 14%, ongoing pregnancies rates were 18% and 14%, and live birth rates were 14% and 12%. Intrauterine injection of hCG via IUI catheter is not recommended in a clinical routine setting at this stage. Future efforts are warranted to further refine the applicability of this modality.

P-586 3D micro-Computed Tomography imaging and reconstruction of the mouse ovary before and after gonadotropins treatment

Abstract Study question How does gonadotropins treatment impact on the number of the different follicle types, their 3D localization, and their recruitment and selection dynamics inside the ovary? Summary answer Treatment with gonadotropins modifies the dynamics of follicles recruitment and selection within the 3D context of the ovary. What is known already Gonadotropins regulate mouse folliculogenesis during follicles recruitment (type 4-5, T4-5) and selection for growth or elimination (T6-7). To further our understanding of the impact of gonadotropins treatment on folliculogenetic dynamics inside the ovary, we used Computed Tomography (CT). 3D imaging of the ovary provides information on its inner spatial organization, and microCT is the only technique that combines high resolution with cubic voxels and allows the organ 3D in-silico reconstruction. We recently published the method for microCT analysis of the adult mouse ovary demonstrating that follicle recruitment occurs simultaneously all-over the cortex, and folliculogenesis is completed within the same region. Study design, size, duration 25-day-old CD1-mice were used since this age rightly precedes the first ovulation: follicles, from the primordial T1 to the pre-ovulatory T8, are all present, but corpora lutea (CL) are yet not formed. Ovaries from different individuals were analyzed by microCT: three not-treated controls (CTR); three injected with 3.5 I.U. PMSG and sacrificed 48hr later (PMSG); three treated first with PMSG, 48hr later injected with 3.5 I.U. hCG and then sacrificed after 15hr upon ovulation (PMSG+hCG). Participants/materials, setting, methods Ovaries were fixed in 4% Paraformaldehyde and treated with Lugol’s solution for 3hr at RT. Then, they were imaged with the EasyTom XL (RX Solutions) tomographic system, using a 1-1.2 µm/pixel resolution. Following microCT imaging, follicles classification (Fiji ImageJ, NIH), 3D mapping (XnConvert, XnSoft) and ovary in-silico reconstruction (Avizo-9, Thermo Fisher Scientific) were performed. ANOVA and Bonferroni post-hoc statistical analyses were done with RStudio &amp;gt; (p &amp;lt; 0.05). Main results and the role of chance Compared to CTR, 48hr after PMSG injection, the T3, T4 and T5 follicle classes remained numerically unchanged, suggesting that the FSH-dependent recruitment involving T4-5 follicles is balanced by an equivalent number of follicles growing from the preceding primordial pool. The most evident change in PMSG ovaries was a 5-fold decrease (p = 0.002) in the number of antral T7 follicles, at the stage of follicle selection. This observation was paralleled by the presence, in the medulla region, of many atretic-like follicles, characterized by a more intense microCT contrast associated with a collapsed antrum, de-structured granulosa-cell layers and fragmented oocytes. In PMSG+hCG ovaries, the number of T3-7 follicles was analogous to that described for PMSG, although the atretic-like follicles almost disappeared, suggesting their elimination sometime prior to ovulation. To understand whether follicle recruitment and selection have a territoriality within the 3D female gonad, ovaries were virtually divided into eight dorsal (D-I/II/III/IV) and ventral (V-I/II/III/IV) sectors. Compared to CTR, PMSG injection appeared to increase follicle selection in sectors V-I (p = 0.03), V-II (p = 0.01) and V-IV (p = 0.02), whereas PMSG+hCG displayed a higher follicle recruitment in sector V-IV (p = 0.0007). Overall, these data suggest a spatial differential effect of the gonadotropins treatment inside the 3D mouse ovary. Limitations, reasons for caution To strengthen the results and better account for the inter-individual variability in response to hormonal treatments, the number of ovaries/individuals should be increased. Also, important is extending the analysis to the time-interval after hCG injection, beyond 15 hr, to assess the effects of ovarian stimulation on subsequent cycles. Wider implications of the findings The method proposed allows to map all follicle stages and to reconstruct a 3D in-silico model of the mouse ovary. This approach could be extended to other Mammals, including human, in normal and pathological conditions, as well as in response to different hormonal stimuli (e.g., dose or type of gonadotropins). Trial registration number not applicable

P-759 Comparison of DNA methylation profiles of human embryo cultured in either uninterrupted or interrupted incubators

Abstract Study question Are there are differences in the DNA methylation profiles of human embryos cultured in Time-lapse imaging (TLI) and standard incubators (SI)? Summary answer The genome-wide DNA methylation landscape was globally similar between the SI and TLI groups. What is known already Early embryonic development is a special biological process, along with dynamic changes of DNA methylation. In vitro culturing ensured the successful development of human embryo, and is the most important step of ART cycle. TLI is one type of newly developed ART embryo culture systems, which allows for continuous assessment of embryonic development. Our previous study performed transcriptome analysis to compare SI and TLI and found that the global transcriptomic profiles were similar between the two groups. However, whether there are differences in the DNA methylation profiles of human embryos cultured in TLI and SI still unknown. Study design, size, duration This study was designed to explore the influence of SI and TLI incubator culturing on genome-wide DNA methylation of human eight-cell embryos. A total of 9 women who received IVF treatment, ≤ 30 years old (range: 20–30 years), without a history of genetic diseases or smoking were included in this study. Participants/materials, setting, methods Transvaginal oocyte retrieval was performed 36 h after HCG injection. Cumulus-enclosed oocytes were collected in 2.5 mL IVF medium and incubated in 5% O2, 6% CO2, and 37 °C incubators for insemination. The fertilized oocytes were transferred into pre-equilibrated Embryoslides. Then the embryoslides were cultured in either TLI or SI at 37 °C with 5% O2 and 6% CO2 until embryo transfer on Day 3. Samples were sequenced by the Illumina HiSeq 4000 with a 150-bp paired-end. Main results and the role of chance The genome-wide methylation patterns and CpG methylation levels in transposable elements and imprinting control regions of TLI-cultured embryos were similar to those of the SI-cultured embryos. However, a small number of differentially methylated regions (DMRs) were detected, and these DMRs mainly occurred in exons other than promoters. Functional annotation revealed that the genes in DMRs tended to execute functions such as cell cycle, DNA damage stimulus, histone modification, mitochondrial, glucose import, and MAPK signaling pathway. Limitations, reasons for caution Small sample size. Wider implications of the findings Evaluated the safety of TLI culture system from the perspective of DNA methylation at single-cell level, and provided an important reference for understanding the association between embryo culture condition and epigenetic regulation. Trial registration number not applicable

The effect of intrauterine hCG injection before embryo transfer on pregnancy rate in frozen embryo transfer cycles

ObjectiveHCG (human chorionic gonadotropin), which is secreted by cytotrophoblast cells, plays an important role in improving pregnancy outcomes among patients with infertility or related problems. In this study, we evaluate the effect of intrauterine hCG injection prior to frozen embryo transfer on pregnancy outcomes. MethodsIn this clinical trial study, among women with infertility problems referred to (XXX) and those with frozen embryos were included in the study. 155 patients in the intervention group received 500 units of hCG while 157 in control group received saline prior to embryo transfer. Along with demographic data, successful in vitro fertilization and clinical pregnancy, loss of pregnancy, successful transplantation, and biochemical parameters were compared among the two groups. ResultsThe mean age of the patients included in the study was 32.97 ± 3.31 years. The level of anti-Mullerian hormone, follicle stimulating hormone and the grade of frozen embryos were not significantly different between the two groups (P > 0.05). The rate of laboratory pregnancy in the intervention group was significantly higher than in the control group (51% vs 35%), p = 0.006. The rate of successful implantation and clinical pregnancy in the intervention group was also significantly higher, p = 0.01 and p = 0.006, respectively. Overall loss of pregnancy in intervention group was 78.1% and 86.0% in control group which was not significantly different, p = 0.068. ConclusionThe outcomes of our study showed that 500 IU of hCG prior to embryo transfer improves the rate of clinical and laboratory pregnancy. However, it does not reduce the rate of loss of pregnancy. Further studies are therefore required in this area.

Effect of the timing of insemination (pre- or post-ovulatory) and type of semen (refrigerated or frozen) on the sex ratio of equine offspring

The normal sex ratio of equine newborns is close to 1:1, with a slight increase of females over males, particularly when the mare is in poor physical condition (Cameron et al., Behavioral Ecology 1999 10(5):472-475). Many other factors like the age of the mare and stallion (Machado Santos et al., Theriogenology 2015 Oct 15;84(7):1238-45), maternal lineage, and uterine status of the mare (Kuhl et al., PLoS ONE 2015, Oct 5;10(10):e0139358) can influence the sex ratio as well. There are some reports describing the impact of insemination timing on the sex of newborns in humans, pigs, and cows (Roelofs et al, Ani Repro Sci. 2006 Jul;93(3-4):366-71; Martin, Hum Reprod. 1996 Aug;11(8):1812-3; Soede et al., Theriogenology 2000 Mar 1;53(4):1003-11). The present work aimed to study the effect of both the timing of insemination and the type of semen (refrigerated versus frozen) on the sex ratio in horse offspring. All the analyzed data were obtained from mares enrolled in a commercial program. In the presence of a preovulatory follicle (>35 mm) and endometrial edema, ovulation was induced by administering 2,500 IU of hCG (IM). Mares intended to be inseminated with refrigerated semen were inseminated after 24 hours from hCG injection, and if they haven't ovulated by then, theywere checked 1 or 2 times a day until ovulation was confirmed. Mares enrolled for insemination with frozen semen were controlled 3 to 4 times a day on the expected day of ovulation. The sex ratio of newborns was not influenced either by the timing of insemination or by the type of semen used (Table 1). Considering our findings, we can conclude that the insemination timing in relation to ovulation and the use of refrigerated or frozen semen, do not affect the sex of offspring in horses.

Evaluation of the association between progesterone levels on the day of HCG injection and the clinical outcomes of IVF treatment cycles in infertile women

Evaluation of the association between progesterone levels on the day of HCG injection and the clinical outcomes of IVF treatment cycles in infertile women

Effects of Acupuncture on Endometrium and Pregnancy Outcomes in Patients with Polycystic Ovarian Syndrome Undergoing in vitro Fertilization-Embryo Transfer: A Randomized Clinical Trial.

To observe the effect of acupuncture on endometrium and pregnancy outcomes in patients with polycystic ovary syndrome (PCOS) infertility undergoing in vitro fertilization-embryo transfer (IVF-ET). Eighty-three patients were randomly assigned to observation group (40 cases) and control group (43 cases) according to the random numbers generated by SPSS software. The patients of the two groups received GnRH agonist long protocol as a routine treatment. In the observation group, acupuncture was given at two acupoint groups for 30 min once every other day. Group 1 included Guanyuan (CV 4), Qihai (CV 6), Zhongji (CV 3), bilateral acupoints Zigong (EX-CA1). Group 2 included Mingmen (GV 4), Yaoyangguan (GV 3), bilateral Shenshu (BL 23) and Ciliao (BL 32). The two groups of acupoints were used alternately. The whole needling process was performed at the time of ovulation induction until the transplantation day and consisted of 3 courses, while the control group did not receive acupuncture interventions. The Gn dosage and Gn stimulation time, endometrial thickness and type (A, B, and C), serum oestradiol (E2) and progesterone (P) levels on the day of injection of human chorionic gonadotropin (hCG), clinical pregnancy rate, as well as live birth rate were observed. Adverse reactions were also be recorded. All patients were followed up for the pregnant rate 14 days after IVF-ET and live birth rate after pregnancy. All adverse reactions (AEs) of acupuncture were recorded during the trial. The Gn dosage and Gn stimulation time in the observation group were lower in the observation group than in the control group (P<0.01). The proportion of type A endometrium in the observation group were higher than that in the control group (P<0.05). The serum E2 and P levels on the day of hCG injection was lower and the clinical pregnancy rate was higher in the observation group compared to those in the control group (P<0.05). There was no serious AEs during this trial. Acupuncture can improve the proportion of type A endometrium, regulate the levels of serum E2 and P on the day of hCG injection, and improve the pregnancy rate in patients with PCOS infertility undergoing IVF-ET.

Kontrollü Ovaryen Hiperstimulasyon Uterusta Apelin ve Apelin Reseptör Ekspresyonunu Arttırır

Objective; In this study, we investigated whether superovulation, which is frequently used in the clinic as controlled ovarian hyperstimulation, affect the expression of Apelin (APLN) and Apelin Receptor (APJ) in the uterus.&#x0D; &#x0D; Methods: Twelve female Balb-C mice 6-8 weeks old were used in this study. The groups were established as follows; virgin control female group (Knt) which did not receive any treatment, the pregnant control group (GK) which was on their pregnancy day 1, and the PMSG+hCG group (P+H) that received 5 IU/mouse/intraperitoneal (ip) PMSG and 5 IU/mouse/ip hCG . Vaginal plaque control was performed on the day after mating in the P+H and GK groups, and vaginal plaque-positive mice were accepted at day 0.5 of pregnancy. Mice in the P+H group were sacrificed 42 hours after hCG injection. Uterine tissues were obtained from all groups for immunohistochemical analysis, and paraffin sections were obtained. Adipose tissue was used as a positive control.&#x0D; &#x0D; Results: In the Knt group, APLN protein expression was present especially in the uterine glands and stromal cells that are located close to the glands. The localization of APJ protein expression was also similar to APLN. In the GK group, APLN and APJ expression levels were similar to the Knt group. However, when the localization of APJ was evaluated, it has been observed that APJ expression was intensely expressed especially in the perimetrial glands of the uterus, which was not present in the Knt group. In the P+H group, APLN expression significantly increased in the endometrial glands and stromal cells compared to the Knt and GK groups (p

Reproductive efficiency of treated Karakul ewes with short-term progesterone and hCG injections during the non-breeding and breeding seasons

The aim of this study was to evaluate reproductive efficiency of ewes following a short-term treatment with progesterone (P4) plus human chorionic gonadotropin (hCG) injections. In Exp 1, ewes (n = 65) were used during the non-breeding season and received P4, three times every other day (n = 22, 3PHCG, IM) or a single dose of P4 (n = 21, PHCG). We injected hCG in all ewes 24 h after the P4 treatment, and distilled water in control group (n = 22). Blood collection were performed on days −13, −6 and + 8 relative to ram release. In Exp 2, during the breeding season, ewes (n = 66) were assigned to the 3PHCG (n = 33) and control (n = 33) groups described for Exp 1, and subsequently received PGF2α 48 h before ram release. Pregnancy diagnosis was performed on day 42 after the ram release which was done 24 h after the hCG injection. In Exp 1, Estrus, pregnancy rate at induced estrus, and lambing rates at the first cycle were greater in 3PHCG (81.8%, 59.1% and 54.5%) group compared with the PHCG (38.1%, 23.8% and 19%) and control (0%, 0% and 0%) groups (P < 0.05). In Exp 2, Estrus, pregnancy rate at induced estrus, and lambing rates at the first cycle were greater in 3PHCG (75.8%, 57.6% and 51.5%) group compared with the control (39.4%, 30.3% and 27.3%) groups (P < 0.05). To conclude, hCG injection 24 h after P4 administration is an effective protocol to induce fertile estrus during non-breeding season, and synchronize estrus and lambing of ewe in the breeding season.

Effect of slightly elevated progesterone on hCG trigger day on clinical pregnancy rate in GnRH-ant IVF/ICSI cycles

BackgroundIt was been agreed that significantly elevated progesterone level on the hCG trigger day have detrimental effect on clinical outcomes in IVF/ICSI cycles. However, few studies explored whether slightly elevated progesterone level also same impact on clinical outcomes.MethodsWe retrospectively studies the effect of slightly elevated progesterone level on outcomes of IVF/ICSI in GnRH-ant cycles. Propensity score matching was used to confounding variables. The women were divided into two groups according to the progesterone level: Group 1: < 1.0 ng/ml; Group 2: 1.0 ng/ml–1.5 ng/ml. Then compare the clinical pregnancy rate (CPR) between the two groups.ResultA total of 847 IVF/ICSI cycles were included in the present study. The average CPR per transfer cycle was 51.7%. CPR of group 1 was 55.22%, significantly higher than that of group 2 (40.66%, P = 0.013). Progesterone level on the day of hCG injection was further evaluated at threshold increments of 0.1 ng/ml, and the CPR was decreased dramatically once the progesterone level higher than 1.4 ng/ml.ConclusionThe slight elevation progesterone level on the hCG trigger day may have a negative effect on the clinical pregnancy in GnRH-ant cycles. In the case of progesterone > 1.4 ng/ml on the hCG injection day, freeze-all strategy was recommended.SummaryThe present retrospective study aimed to evaluate the effect of slightly elevated progesterone (1.0 ng/ml ~ 1.5 ng/ml) on outcomes of IVF/ICSI in GnRH-ant cycles. Slightly elevated progesterone level leaded to significant lower clinical pregnancy rate (CPR) that that of group with normal progesterone level (40.66% vs. 55.22%, P = 0.013). The CPR was decreased dramatically once the progesterone level higher than 1.4 ng/ml. So slightly elevated progesterone level on the trigger day may have a negative effect on the clinical pregnancy in GnRH-ant cycles. In the case of progesterone > 1.4 ng/ml on the hCG injection day, freeze-all strategy was recommended.

Efficacy and safety of newly developed ganirelix acetate in infertile women for assisted reproductive technology: a prospective, randomised, controlled study

This study aimed to investigate the efficacy of Ganilever pre-filled syringe (PFS), a newly developed ganirelix acetate, for the inhibition of premature luteinising hormone (LH) surge in in vitro fertilisation (IVF). A prospective randomised controlled study was conducted (NCT03051087). A total of 236 women (Ganilever group: 114, Orgalutran group: 122) were finally analysed. The patients with LH of >10 mIU/mL on the day of human chorionic gonadotropin (hCG) injection were 0 (0.0%) and 3 (2.5%) in the Ganilever and Orgalutran groups, respectively (p= .25). The number of retrieved oocytes from two groups did not show any significant difference (12.0 ± 6.4 vs. 11.8 ± 6.3, p= .73). Furthermore, the two groups did not show significant differences in the number of good-quality oocytes and embryo, and the rate of fertilisation. Similar safety profiles were also observed. In conclusion, Ganilever PFS showed comparable IVF outcomes and safety profile in IVF, as compared to the Orgalutran. Impact Statement What is already known on this subject? Premature LH surge during controlled ovarian stimulation results in the induction of luteinisation of the immature follicles. Thus, gonadotrophin-releasing hormone (GnRH) antagonist protocol was suggested as an option for suppression of premature LH surge. Currently, one of GnRH antagonists being widely used is ganirelix acetate (Orgalutran®; Organon, Oss, The Netherlands). Ganilever pre-filled syringe (PFS) is a newly developed GnRH antagonist containing ganirelix acetate as an active ingredient. What do the results of this study add? Our study demonstrated that Ganilever PFS showed comparable IVF outcomes and patient safety profile in infertile women undergoing in IVF-ET, as compared to the Orgalutran. What are the implications of these findings for clinical practice and/or further research? The results of our study will provide another available GnRH antagonist to be used in patients with IVF.

The Effects of Prostaglandin E2 Treatment on the Secretory Function of Mare Corpus Luteum Depends on the Site of Application: An in vivo Study.

We examined the effect of prostaglandin (PG) E2 on the secretory function of equine corpus luteum (CL), according to the application site: intra-CL injection vs. an intrauterine (intra-U) administration. Moreover, the effect of intra-CL injection vs. intra-U administration of both luteotropic factors: PGE2 and human chorionic gonadotropin (hCG) as a positive control, on CL function was additionally compared. Mares were assigned to the groups (n = 6 per group): (1) an intra-CL saline injection (control); (2) an intra-CL injection of PGE2 (5 mg/ml); (3) an intra-CL injection of hCG (1,500 IU/ml); (4) an intra-U saline administration (control); (5) an intra-U administration of PGE2 (5 mg/5 ml); (6) an intra-U administration of hCG (1,500 IU/5 ml). Progesterone (P4) and PGE2 concentrations were measured in blood plasma samples collected at −2, −1, and 0 (pre-treatment), and at 1, 2, 3, 4, 6, 8, 10, 12, and 24 h after treatments. Moreover, effects of different doses of PGE2 application on the concentration of total PGF2α (PGF2α and its main metabolite 13,14-dihydro-15-keto-prostaglandin F2α– PGFM) was determined. The time point of PGE2, hCG, or saline administration was defined as hour “0” of the experiment. An intra-CL injection of PGE2 increased P4 and PGE2 concentrations between 3 and 4 h or at 3 and 12 h, respectively (p < 0.05). While intra-U administration of PGE2 elevated P4 concentrations between 8 and 24 h, PGE2 was upregulated at 1 h and between 3 and 4 h (p < 0.05). An intra-CL injection of hCG increased P4 concentrations at 1, 6, and 12 h (p < 0.05), while its intra-U administration enhanced P4 and PGE2 concentrations between 1 and 12 h or at 3 h and between 6 and 10 h, respectively (p < 0.05). An application of PGE2, dependently on the dose, supports equine CL function, regardless of the application site, consequently leading to differences in both P4 and PGE2 concentrations in blood plasma.

COMPARISON OF SINGLE DOSE AND MULTI-DOSE hCG STIMULATION TESTS

Objective: The human chorionic gonadotropin (hCG) stimulation test is widely used to assess testicular steroidogenesis. This test evaluates Leydig cell function and helps to evaluate testosterone synthesis in testicular enzyme defects, hypogonadism, 46,XY sexual development disorders (DSD), and micropenis. This study will compare testosterone responses to a single dose of 5000 IU/m2 and three consecutive days of 1500 IU/m2 hCG regimens. Materials and Methods: The study evaluated 18 patients who were admitted with micropenis, hypospadias, or undescended testis (Group 1) and 18 patients with a molecular diagnosis of androgen receptor insensitivity (56%) or 5α reductase deficiency (44%) (Group 2). The median (interquartile range; IQR) age of the patients were 1.4 (0.8-4.2) years for Group 1 and 0.7 (0.3-0.8) years for Group 2. Baseline testosterone levels were checked and after a single dose of hCG 5000 IU/m2 injection into the first group, testosterone levels were observed on day one and day four. In the second group, after the injection of hCG 1500 IU/m2 for three days, testosterone levels were checked on the fourth day. Results: There was no significant difference in height, weight, BMI SDS, and baseline testosterone levels between groups. After hCG, stimulation testosterone responses did not vary among the groups. Testosterone levels were increased 40-fold and 40.1- fold on day four compared to the baseline in Group 1 and Group 2. In Group 1, testosterone response was significantly higher on day four compared to day one (p=0.001). Conclusions: The hCG tests performed with a single dose or multidoses showed that there was no difference between testosterone responses and these two tests were applicable the 5000 IU/m2 hCG test requires fewer injections and is easy to administer, this test may be preferred to an evaluation of the Leydig cell function. For an adequate testosterone response, the researchers suggest measuring testosterone levels on the fourth day after the single dose of 5000 IU/m2 .

Prevention of Premature Ovulation by Administration of Gonadotropin Releasing Hormone Antagonist the day After Ovulation Triggering in Diminished Ovarian Reserve Patients.

The aim of the present retrospective study was to investigate the effectiveness of single-dose gonadotropin releasing hormone (GnRH) antagonist administration, the day after human chorionic gonadotropin (hCG) triggering for final oocyte maturation, on the prevention of premature luteinization in patients with diminished ovarian reserve in in-vitro fertilization (IVF) cycles. The secondary objective of the study was to search the effect of this protocol on pregnancy outcomes. This is a retrospective study including 267 infertile patients who have single antral follicle seen with ultrasonography on the 2nd or 3rd day of the menstrual cycle before starting IVF treatment. We randomized patients into two groups. The case group comprised patients who had single-dose GnRH antagonist injection the day after hCG triggering formed, and the patients who had the standard treatment regime formed the control group. In both groups, the oocytes were collected 36 hours after hCG injection. The premature ovulation rate was significantly low in the case group compared with the control group (6.86 versus 20.6% per scheduled cycle) (p = 0.022). Also, the oocyte retrieval rate (93.14 versus 67.87% per scheduled cycle) (p = 0.013), the oocyte maturity rate (79.42 versus 47.87%) (p = 0.041), the fertilization rate (65.68 versus 34.54%) (p = 0.018), and the embryo transfer rate per scheduled cycle (44.11 versus 18.78%) (p = 0.003) were higher in the GnRH antagonist group than in the control group. The administration of GnRH antagonist the day after hCG trigger in IVF treatments of patients with diminished ovarian reserve enabled a significant decrease in the rate of premature ovulation but had no effect on live birth rate.

Clinical aspects of oocyte retrieval and embryo transfer: tips and tricks for the novice and the expert.

Oocyte retrieval (ovum pick-up) and embryo transfer (ET) are essential steps in in-vitro fertilization and intracytoplasmic sperm injection and over the years, the two procedures were developed in order to improve the clinical outcome. Many suggestions were proposed and applied before, during and after oocyte retrieval such as timing of HCG trigger, pre-operative pelvic scan, vaginal cleansing, type of anesthesia, type and gauge of aspiration needles, aspiration pressure, follicle flushing, and the need for prophylactic antibiotics. Similarly, many steps were suggested and implemented before, during and after ET including patient's position, type of anesthesia/analgesia, dummy (mock) ET, ultrasound-guidance, HCG injection in the uterine cavity, use of relaxing agents, full bladder, removal of the cervical mucus, flushing the cervix with culture medium, type of ET catheter, embryo loading techniques, site of embryo deposition, the use of adherence compounds, as well as bed rest after ET. Complications were also reported with oocyte retrieval and ET. The aim of this review is to evaluate the current practice of these two procedures in the light of available evidence.

Ultrasonographic and macroscopic study of pregnancy in golden hamster

BackgroundHamster is widely used as an experimental model in the study of reproductive system. However, pregnancy diagnosis and aging always have been a challenge. ultrasonography have been used in diagnosis of pregnancy in some small laboratory animals, such as rabbits, rats, and mice. Current study describes use of trans-abdominal ultrasonography for pregnancy diagnosis and fetal age estimation in golden hamster. Furthermore, a macroscopic examination was performed to evaluate the embryonic vesicle diameter, crown-rump length, and fetal head diameter. Ten adult female golden hamsters were selected and maintained under controlled light conditions (14 h light/10 h darkness). The estrous cycle was synchronized using eCG and hCG. During estrous (18 h after hCG injection), the hamsters were naturally mated. After seven days of mating, the hamsters were examined daily for pregnancy diagnosis and aging with an ultrasound scanner equipped with an 8.5-MHZ linear probe. On each day of the experiment, at least one of the pregnant hamsters was euthanized and dissected for macroscopic fetal measurements using a digital caliper.ResultsThe gestational sac and crown-rump length were identified and measured by ultrasonographicly on day 7 of pregnancy and head could be visible after day 10 of gestation. Statistical analysis revealed that the ultrasound estimation of gestational age was significantly correlated with the actual age of the fetus (r = 0.98; p < 0.05).ConclusionsReal-time ultrasound can be used for the diagnosis of pregnancy and estimation of fetal age in golden hamster from day 7 of gestation.

Effect of GnRH and hCG injection on the reproductive performance and serum progesterone concentration of ewes during spring season.

This study was carried out to investigate the effect of different hormonal programs on the reproductive parameters and serum progesterone concentration of Lake-Ghashghaei ewes in spring season. In this study, 144 ewes were selected from two large nomadic herds. The ewes were synchronized using progesterone sponges. One day before harvesting the sponges, all ewes were injected intramuscularly with 400 IU of eCG. Then ewes were divided into six groups (12 ewes). In the first herd, 1 (GnRH-1 group), 2 (GnRH-2 group), 5 (GnRH-5 group), 7 (GnRH-7 group) and 12 days (GnRH-12 group) after estrus observation, 25.00 µg GnRH was injected intramuscularly to the ewes. In the second herd, the ewes were received 500 IU of hCG on days 1 (hCG-1), 2 (hCG-2), 5 (hCG-5), 7 (hCG-7) and 12 (hCG-12). In each herd, a group of 12 ewes received only eCG and progesterone sponges and was considered as control group. The results showed that twinning rate, the number of born lambs, fecundity and prolificacy rate were higher GnRH and hCG received groups on the second day compared to other groups. The serum progesterone concentration was significantly higher in all groups received GnRH and hCG compared to control group. In general, the results showed that GnRH and hCG two days after mating improved different reproductive activities in addition to increasing serum progesterone concentration. On the other hand, hCG had a greater effect on progesterone concentration, the number of born lambs and the fecundity rate compared to GnRH.

Timed Intercourse versus Intrauterine Insemination with Mild Ovarian Stimulation for Unexplained Infertility

Background: Infertility is unexplained in about 22% - 28% of infertile couples. Spontaneous pregnancy may occur in them. If did not happen, Expectant management will be via clomiphene citrate (CC) administration, intrauterine insemination (IUI), and IVF & ICSI. Aim: to assess the effectiveness of intrauterine insemination with mild controlled ovarian stimulation compared with expectant management in couples with unexplained infertility more than one year. Methods: 160 couples with unexplained infertility were selected, The couples were randomly divided into two groups: Group (A) “80 couples”: Intrauterine insemination (IUI) with mild controlled ovarian stimulation (combination of CC + hMG) using prepared semen and was performed 36 hours after hCG injection, Group (B) “80 couples”: Couples had no ovulation induction. They encouraged for timed intercourse in the most fertile days of female cycle guided by folliculometry for 6 months. Data were collected quantitatively, coded and analyzed using SPSS. The power of study is 80% and 95% confidence interval. Result: The pregnancy rate was calculated in both groups: In group (A) (IUI/COS): OPR (Ongoing pregnancy rates) was 27.5% and PR (pregnancy rates)/cycle was 8.6%. In group (B) (Expectant management): OPR was 25% and PR/cycle was 5.3%. Conclusion: Mild controlled ovarian stimulation CC + hMG with IUI offers no statistical significance in terms of pregnancy outcomes over expectant management in this study however significance in the response to stimulation.