hCG Injection Research Articles

Evaluation the effect of sildenafil on nitric oxide secretion and improvement of endometrial receptivity in fresh ICSI cycles

Introduction: Endometrial receptivity is a chain of events that promotes embryo implantation and is regulated by a variety of substances, such as cytokines, growth factors, and steroid hormones. The study's objective is to assess how vaginally delivered sildenafil in ICSI patients affects pregnancy outcome, endometrial thickness, subendometrial resistance index, and pulsatility index. Methods: The sixty women who participated in this controlled randomised trial (interventional experimental research) at the High Institute of Infertility Diagnosis and Assisted Reproductive Technologies underwent IVF/ICSI cycles. Patients were split into two groups: Study group: From the day of the last menstrual period to the day of the hCG injection, thirty (30) infertile women got four (4) doses of vaginal sildenafil 25mg tablets daily. Thirty (30) infertile ladies who weren't given sildenafil therapy made up the control group. Results: Nitric oxide levels in the study group's serum were significantly significant. In the subendometrial Doppler research, the mean pulsatility index was considerably lower in the study group whereas the mean resistance index and mean systolic to diastolic ratio did not change statistically between the control and study groups. Between the control and study groups, there was no statistically significant difference in the mean endometrial thickness. The sildenafil group had a greater pregnancy rate, although the difference was not statistically significant. Conclusion: By enhancing subendometrial blood flow by lowering the pulsatility index and raising the nitric oxide (NO) factor, the use of vaginal sildenafil in a new ICSI cycle improves endometrial receptivity.

Application of ultrasound markers measured at different time points of COH cycle in the prediction of ovarian response for individualised ovulation induction

The purpose of this study was to evaluate the predictive value of ultrasound markers measured at different time points of the controlled ovarian hyperstimulation (COH) cycle on ovarian response and outcome indicators in the IVF-ET cycle. According to the oestrogen level and the number of retrieved oocytes, patients who planned for COH treatment were separated into low-response group, normal and high-response group. The ovarian stromal artery flow parameters on the day of pituitary down-regulation, day 1, day 7, day 10, and the day of hCG injection were collected prospectively. We also have collected the data of cumulus oophorus count on the day of hCG injection by transvaginal sonography. Compared with the low-response group, on the first day of the COH cycle PI, RI, and S/D were lower in the high-response group than they were in the low-response group (p < .05). PSV and EDV were significantly higher in the high-response group than they were in the low-response group (p < .01), and the PSV on the first day of the COH cycle have statistical significance in predicting the number of high-quality embryos. The number of cumulus oophorus on the day of hCG injection has statistical significance in predicting the number of oocytes retrieved and fertilised oocytes. We conclude that the ovarian stromal artery flow parameters on the first day of the COH cycle and cumulus oophorus count on hCG injection day can serve as efficient indicators for an early assessment of ovarian response and individualised ovulation induction. IMPACT STATEMENT What is already known on this subject? AMH, AFC, and the age of the patient are well-known effective parameters for the evaluation of ovarian response, but these are insufficient and full of individual differences. Some researchers have investigated the value of colour Doppler ultrasound and cumulus oophorus in assessing ovarian response, but no definitive conclusion has been reached. What do the results of this study add? The hemodynamic parameters of ovarian stromal artery on the first day of the COH cycle and the number of cumulus oophorus on the day of hCG injection detected by Transvaginal Colour Doppler Sonography (TV-CDS) could be used to predict the ovarian response. What are the implications of these findings for clinical practice and/or further research? Ovarian stromal artery flow parameters and cumulus oophorus detected by TV-CDS can potentially be offered as a complementary parameter for ovarian reserve.

The impact of maternal age on gene expression during the GV to MII transition in euploid human oocytes.

STUDY QUESTIONAre there age-related differences in gene expression during the germinal vesicle (GV) to metaphase II (MII) stage transition in euploid human oocytes?SUMMARY ANSWERA decrease in mitochondrial-related transcripts from GV to MII oocytes was observed, with a much greater reduction in MII oocytes with advanced age.WHAT IS KNOWN ALREADYEarly embryonic development is dependent on maternal transcripts accumulated and stored within the oocyte during oogenesis. Transcriptional activity of the oocyte, which dictates its ultimate developmental potential, may be influenced by age and explain the reduced competence of advanced maternal age (AMA) oocytes compared with the young maternal age (YMA). Gene expression has been studied in human and animal oocytes; however, RNA sequencing could provide further insights into the transcriptome profiling of GV and in vivo matured MII euploid oocytes of YMA and AMA patients.STUDY DESIGN, SIZE, DURATIONFifteen women treated for infertility in a single IVF unit agreed to participate in this study. Five GV and 5 MII oocytes from 6, 21–26 years old women (YMA cohort) and 5 GV and 6 MII oocytes from 6, 41–44 years old women (AMA cohort) undergoing IVF treatment were donated. The samples were collected within a time frame of 4 months. RNA was isolated and deep sequenced at the single-cell level. All donors provided either GV or MII oocytes.PARTICIPANTS/MATERIALS, SETTING, METHODSCumulus dissection from donated oocytes was performed 38 h after hCG injection, denuded oocytes were inserted into lysis buffer supplemented with RNase inhibitor. The samples were stored at −80°C until further use. Isolated RNA from GV and MII oocytes underwent library preparation using an oligo deoxy-thymidine (dT) priming approach (SMART-Seq v4 Ultra Low Input RNA assay; Takara Bio, Japan) and Nextera XT DNA library preparation assay (Illumina, USA) followed by deep sequencing. Data processing, quality assessment and bioinformatics analysis were performed using source-software, mainly including FastQC, HISAT2, StringTie and edgeR, along with functional annotation analysis, while scploid R package was employed to determine the ploidy status.MAIN RESULTS AND THE ROLE OF CHANCEFollowing deep sequencing of single GV and MII oocytes in both YMA and AMA cohorts, several hundred transcripts were found to be expressed at significantly different levels. When YMA and AMA MII oocyte transcriptomes were compared, the most significant of these were related to mitochondrial structure and function, including biological processes, mitochondrial respiratory chain complex I assembly and mitochondrial translational termination (false discovery rate (FDR) 6.0E−10 to 1.2E−7). These results indicate a higher energy potential of the YMA MII cohort that is reduced with ageing. Other biological processes that were significantly higher in the YMA MII cohort included transcripts involved in the translation process (FDR 1.9E−2). Lack of these transcripts could lead to inappropriate protein synthesis prior to or upon fertilisation of the AMA MII oocytes.LARGE SCALE DATAThe RNA sequencing data were deposited in the Gene Expression Omnibus (https://www.ncbi.nlm.nih.gov/geo), under the accession number: GSE164371.LIMITATIONS, REASONS FOR CAUTIONThe relatively small sample size could be a reason for caution. However, the RNA sequencing results showed homogeneous clustering with low intra-group variation and five to six biological replicates derived from at least three different women per group minimised the potential impact of the sample size.WIDER IMPLICATIONS OF THE FINDINGSUnderstanding the effects of ageing on the oocyte transcriptome could highlight the mechanisms involved in GV to MII transition and identify biomarkers that characterise good MII oocyte quality. This knowledge has the potential to guide IVF regimes for AMA patients.STUDY FUNDING/COMPETING INTEREST(S)This work was supported by the Medical Research Council (MRC Grant number MR/K020501/1).

Measuring the Effects of a Single Dose of Human Chorionic Gonadotropin (hCG) on Plasma Testosterone Concentrations in Leopard Geckos (Eublepharis macularius)

Current estimates suggest that one-fifth of all reptiles are currently threatened with extinction because of anthropogenic activity, and that if nothing is done to mitigate these impacts, massive extinctions will occur over the coming decades. Assisted reproductive technologies (ART) can be used to develop ex vivo conservation programs to ensure species survival until environmental deficiencies can be corrected. Geckos represent one of the most diverse groups of reptiles on earth and need protection; however, to date, no ART have been developed for these animals. Exogenous human chorionic gonadotropin (hCG) is routinely used to alter reproductive hormones of vertebrates and are integrated in ART. The purpose of this study was to determine if a single dose of hCG could increase plasma testosterone concentrations in the leopard gecko (Eublepharis macularius) within 24 h of administration. The study was performed in September, which is considered the nonbreeding season for this species in the northern hemisphere. Twelve captive-bred, adult, male leopard geckos were randomly divided into three treatment groups and administered an intramuscular injection of hCG (50 or 100 IU) or saline. Whole blood was collected prior to injection (baseline) and 12 and 24 h after injection. Plasma testosterone concentrations were measured using an enzyme immunoassay kit. There was no significant difference (P = 0.195) in testosterone concentrations between treatment groups. Because there was no difference, an observed testosterone range (61.0 ± 42.5, 5.5–149.0) for September was calculated. These results suggest that captive leopard geckos may be in a recrudescence phase in September in the northern hemisphere.

Role of Adding Phytoestrogen to Clomiphene Citrate in Improving Endometrial Thickness in Woman with Unexplained Infertility

Background: Unexplained infertility is failure to conceive after 1 year in couples with normal semen samples and no abnormality found during an infertility work-up. Clomiphene citrate (CC) induces ovulation. Objective: The aim was to evaluate the effect of follicular phase oral phytoestrogens (PES) in patients with unexplained infertility, managed by clomiphene ovulation induction, triggered by HCG injection and timed intercourse, in terms of pregnancy rate, follicular maturation and endometrial thickness. Patients and Methods: This study included 56 infertile patients seeking for pregnancy. Clomiphene was given orally at a dose of one tablet twice per day (tablet 50 mg), from day 3 to day 7 for all patients (group I and group II), while phytoestrogens were given at a dose of two tablets one shot per day (tablet 20 mg), from day 1 to day 12 only for group I, followed by HCG injection on attaining mature follicle(s) by trans-vaginal US scan with diameter ≥ 18 mm in both group. The study was conducted in Department of Obstetrics and Gynecology, Zagazig University Hospitals. Results: The number of days till HCG injection were significantly lower in group I (13.8 ± 1.8 days) than in group II (14.9 ± 2.1 days), which indicates a better ovulation. As for the endometrial thickness, it was significantly higher in group I than in group II. The number of ovulatory follicle(s) as single, two or three showed no significant difference between the studied groups. Conclusion: Adding PES to CC induced cycles, improved the endometrial thickness, promoted follicular maturation in a shorter time and also, improved pregnancy rates.

VP07.05: Relationship between serum progesterone levels and endometrial status on hCG injection's day in ART cycles

VP07.05: Relationship between serum progesterone levels and endometrial status on hCG injection's day in ART cycles

Dynamic Oxygen Conditions Promote the Translocation of HIF-1α to the Nucleus in Mouse Blastocysts.

Oxygen tension is one of the most critical factors for mammalian embryo development and its survival. The HIF protein is an essential transcription factor that activated under hypoxic conditions. In this study, we evaluated the effect of dynamic oxygen conditions on the expression of embryonic genes and translocation of hypoxia-inducible factor-1α (HIF-1α) in cultured mouse blastocysts. Two-pronuclear (2PN) zygotes harvested from ICR mice were subjected to either high oxygen (HO; 20%), low oxygen (LO; 5%), or dynamic oxygen (DO; 5% to 2%) conditions. In the DO group, PN zygotes were cultured in 5% O2 from days 1 to 3 and then in 2% O2 till day 5 after hCG injection. On day 5, the percentage of blastocysts in the cultured embryos from each group was estimated, and the embryos were also subjected to immunocytochemical and gene expression analysis. We found that the percentage of blastocysts was similar among the experimental groups; however, the percentage of hatching blastocysts in the DO and LO groups was significantly higher than that in the HO group. The total cell number of blastocysts in the DO group was significantly higher than that of both the HO and LO groups. Further, gene expression analysis revealed that the expression of genes related to the embryonic development was significantly higher in the DO group than that in the HO and LO groups. Interestingly, HIF-1α mRNA expression did not significantly differ; however, HIF-1α protein translocation from the cytoplasm to the nucleus was significantly higher in the DO group than in the HO and LO groups. Our study suggests that dynamic oxygen concentrations increase the developmental capacity in mouse preimplantation embryos through activation of the potent transcription factor HIF-1α.

Ankyrin-repeat and SOCS box-containing protein 9 (ASB9) regulates ovarian granulosa cells function and MAPK signaling.

Ankyrin-repeat and SOCS box-containing proteins (ASB) interact with the elongin B-C adapter via their SOCS box domain and with the cullin and ring box proteins to form E3 ubiquitin ligase complexes within the protein ubiquitination pathway. ASB9 in particular is a differentially expressed gene in ovulatory follicles (OFs) induced by the luteinizing hormone (LH) surge or hCG injection in ovarian granulosa cells (GC) while downregulated in growing dominant follicles. Although ASB9 has been involved in biological processes such as protein modification, the signaling network associated with ASB9 in GC is yet to be fully defined. We previously identified and reported ASB9 interactions and binding partners in GC including PAR1, TAOK1, and TNFAIP6/TSG6. Here, we further investigate ASB9 effects on target binding partners regulation and signaling in GC. CRISPR/Cas9-induced inhibition of ASB9 revealed that ASB9 regulates PAR1, TAOK1, TNFAIP6 as well as genes associated with proliferation and cell cycle progression such as PCNA, CCND2, and CCNE2 while CCNA2 was not affected. Inhibition of ASB9 was also associated with increased GC number and decreased caspase3/7 activity, CASP3 expression, and BAX/BCL2 ratio. Furthermore, ASB9 induction in OF in vivo 24 h post-hCG is concomitant with a significant decrease in phosphorylation levels of MAPK3/1 while pMAPK3/1 levels increased following ASB9 inhibition in GC in vitro. Together, these results provide strong evidence for ASB9 as a regulator of GC activity and function by modulating MAPK signaling likely through specific binding partners such as PAR1, therefore controlling GC proliferation and contributing to GC differentiation into luteal cells.

TARGETED DELETION OF MITOFUSIN 1 AND MITOSUFIN 2 CAUSES FEMALE INFERTILITY AND LOSS OF FOLLICULAR RESERVE

Mitofusin 1 (MFN1) and Mitofusin 2 (MFN2) are transmembrane GTPases that regulate mitochondrial fusion and are required for the maintenance of cellular homeostasis. The aim of the study was to determine the role of mitofusins in female reproductive competence and senescence using a mouse model with oocyte-specific deletion of Mfn1 and Mfn2. Mice with oocyte-specific double deletion of Mfn1 and Mfn2 were generated by mating Mfn1 flox/flox or Mfn2 flox/flox with Zp3-Cre Mfn1 and Mfn2 males. In all experiments, adult (8-weeks-old) Mfn1-/-/Mfn2-/- female mice were compared to wild-type (WT). To assess fertility, female mice from each group (n=5) were mated with adult WT males of proven fertility for 12 weeks. The ability to generate oocytes (germinal vesicle [GV] and metaphase II [MII]) was assessed after injection with PMSG (10IU) or PMSG and hCG (10IU). Unfertilized MII oocytes were collected 14 hours after hCG injection. Follicle development was assessed in ovaries from Mfn1-/-/Mfn2-/- and WT mice after fixation, paraffin embedding, and sectioning, followed by hematoxylin and eosin (H&E) staining. ATP levels were determined by bioluminescent assay. Mitochondrial DNA (mtDNA) copy number was measured in individual oocytes by cloning of mitochondria specific gene (Cox3) as a standard, followed by quantitative real-time PCR (qPCR). Expression levels of mitochondrial respiratory chain genes was evaluated with qRT-PCR. Student’s t-test and Chi-Square analysis were applied. Mature female Mfn1-/-/Mfn2-/- were infertile compared to WT females (0 ± 0 vs ± 5.62 ± 1.64 pups per litter; p < 0.0001). Mfn1-/-/Mfn2-/- mice showed a significant decrease in GV oocyte number (12.75 ± 1.70 vs 40.25 ± 3.5; p < 0.0001) and generated no MII oocytes (0 ± 0 vs 31.01 ± 3.5; p < 0.0001) in comparison to WT. Eight-week-old Mfn1-/-/Mfn2-/- mice ovaries had similar number of primordial, primary, and secondary follicles compared to WT. However, the number of early antral and antral follicles numbers was significantly decreased (17.2 ± 10.2 vs 55.3 ± 9.50; p < 0.03, and 2.3 ± 1.5 vs 16.3 ± 5.8; p < 0.04). At 6 months, Mfn1-/-/Mfn2-/- mice had a lower number of primordial, early antral, and antral follicles compared to WT. At 12 months, all follicle stages were found to be significantly lower in Mfn1-/-/Mfn2-/- mice. Decreased ATP production (0.62 ± 0.1 vs 1.23 ± 0.12; p = 0.002) and mtDNA copy number (446033 ± 53978 vs 706672 ± 72470; p = 0.003) were also observed in Mfn1-/-/Mfn2-/-oocytes compared to WT. Similarly, the expression of mitochondrial chain respiratory genes Cox1, Sdhb, Uqcrc2 and Atp5a1 were decreased in Mfn1-/-/Mfn2-/- oocytes compared to WT. Our findings demonstrate that lack of MFN1 and MFN2 results in infertility and accelerated loss of follicular reserve consistent with a diminished ovarian reserve (DOR) phenotype.

Validation of the existence of genuine Empty follicle syndrome, versus false empty follicle syndrome to make definitive decisions in cases where recurrent IVF failure encountered secondary to absence of oocytes on ovum pick up-a short communication

Aim: Worldwide a big altercation exists with regards to the actual existence of the term “ Empty follicle syndrome’’(EFS), so much so that certain big authorities in the field have been believing that true EFS does not exist. Basically EFS is a syndrome when no functionally intact oocyte get retrieved when attempting an oocyte pick up (OPU) for a successful in vitro fertilization (IVF), however such patients encounter recurrent IVF failures. Since it is has become a big problem for the treating reproductive endocrinologist, besides the patient encountering recurrent IVF failures, it has become essential to differentiate the true EFS alias genuine Empty follicle syndrome (gEFS) from what is labeled today as the false empty follicle syndrome (fEFS). In view of the recently documented presence of mutations, gEFS got verified and appears to silence this biggest conflict that arose secondary to the existence of a false empty follicle syndrome(fEFS), where one could manage to get successful IVF outcomes subsequent to repeated hCG injections/ gonadotropin releasing hormone (GnRH) agonist in addition to pregnancy, with lot of clinicians believing there is no true term like EFS. Methods: Recently Yang et al., performed a study In tertiary a university based reproductive center in China that was comprised of a big cohort of patients that presented with gEFS. Genetic evaluation was conducted on 35 non correlated infertile patients who went through 16 failed IVF cycles in addition to oocyte degeneration, besides the subjects got a diagnosis of possessing a particular kind of EFS- cumulus oocytes complexes (COC’s) but possessed oocytes that were undergoing degeneration, with the utilization of whole –exome sequencing along with targeted Sanger sequencing. Results: Yang et al., found 22 innovative genetic variant of zona pellucida (ZP), genes in 18 subjects, that were inclusive of 20 variants in ZP 1 gene, 2 in ZP 2 gene in addition to 1 recurring variant in ZP3 gene that had been earlier documented. The homogenous /compound heterogenous ZP 1 mutations were inherited in an autosomal recessive manner, while the heterogenous variants of ZP 2 as well as ZP3 genes possessed an autosomal dominant manner of inheritance. Conclusions: These mutations were anticipated to be harmful in silico along with got further experimentally corroborated to be functionally null dependent on their ectopic expression in vitro. Thus with this further evidence that has been recently provided with regards to the existence of genuine Empty follicle syndrome (gEFS), it is significant for the youngsters to realize if they encounter similar cases after trial of rescue hCG injections / GnRH) agonist, not to further keep waiting, but evaluate further with regards to the existence of mutations for Zona Pellucida (ZP), ZP 1, ZP2 as well as ZP3 genes mutations, or LH/ chorionic gonadotropins receptor (LHCGR) gene mutation without subjecting the patient to repeated IVF, with her psychological as well as financial health in mind

LH Induces De Novo Cholesterol Biosynthesis via SREBP Activation in Granulosa Cells During Ovulation in Female Mice.

In the liver, the sterol response element binding protein (SREBP) and the SREBP cleavage-activated protein (SCAP) complex upregulate cholesterol biosynthesis by gene induction of de novo cholesterol synthetic enzymes (Hmgcr, Cyp51, and Dhcr7). Insulin induced gene 1 (INSIG1) negatively regulates cholesterol biosynthesis by the inhibition of de novo cholesterol biosynthetic gene expression. In the ovary, cholesterol is de novo synthesized; however, the roles of SREBP and its regulators (SCAP and INSIG1) are not well understood. In this study, when immature mice were treated with gonadotropins (eCG followed by hCG), eCG induced and hCG maintained the expression of SREBP-1a, -2, and SCAP granulosa cells, whereas INSIG1 expression was dramatically downregulated after hCG injection. Downregulation of INSIG1 led to generate the SREBPs active form and translocate the SREBPs active form to nuclei. Inhibition of generation of the SREBPs active form by fatostatin or Scap siRNA in both in vivo and in vitro significantly decreased the expressions of de novo cholesterol biosynthetic enzymes, cholesterol accumulation, and progesterone (P4) production compared with the control group. Fatostatin treatment inhibited the ovulation and increased the formation of abnormal corpus luteum which trapped the matured oocyte in the corpus luteum; however, the phenomenon was abolished by P4 administration. The results showed that decreasing INSIG1 level after hCG stimulation activated SREBP-induced de novo cholesterol biosynthesis in granulosa cells of preovulatory follicles, which is essential for P4 production and the rupture of matured oocyte during ovulation process.

Angka kebuntingan sapi perah dengan korpus luteum persisten setelah pemberian kombinasi prostaglandin F2α dan gonadotropin

This case study aims to determine the effect of a combination of prostaglandin F2α (PGF2α) with Gonadotropin (PG-600), and human chorionic gonadotropin (hCG) on the estrus and pregnancy rates in Holstein Friesian (HF) cows with persistent corpus luteum (PCL). A total of 15 Holstein Friesian (HF) dairy cows aged 3-5 years with body condition score (BCS) 5-7 (scale 1-9) diagnosed with PCL, were injected with 7.5 mg PGF2α intra-vulvar submucosa. Cows were then divided into three treatment groups. Group P0 was not given the combination treatment, while P1 group was injected with a combination of 300 IU PG-600 and 150 IU hCG, meanwhile P2 group was was injected with a combination of 300 IU PG-600 and 300 IU hCG intra-muscularly. The PGF2α and PG-600 injections were carried out on the same day PCL was diagnosed. Meanwhile, the injection of hCG was carried out at the same time as the AI. The Pregnancy examination was performed 90 days after AI through rectal palpation. The results showed that all treated cows were estrus observed with the fastest onset of estrus was the P2 group, that was significantly different (p &lt;0.05) compared to the P0 and P1 groups. All treated cows were diagnosed as pregnant. It can be concluded that the administration of PGF2α or its combination with gonadotropins in HF cows with PCL could result in 100% estrus and pregnancy rates.

Therapy of Human Chorionic Gonadotropin (hCG) Hormone in Dairy Cattles that Have Repeat Breeder on Pregnancy Rate in Tulungagung, East Java, Indonesia

This study aimed to prove that giving hCG injection at the time of artificial insemination and five days afterartificial insemination in dairy cows with repeat breeder can cause pregnancy. This study employed 30 dairycows with repeat breeder. All samples of dairy cows were divided into two treatments, P1 using hCG at adose of 100 IU given intramuscularly at the time of artificial insemination and P2 using hCG at a dose of100 IU intramuscularly given five days after artificial insemination. The results were analyzed using the Chi-Square test. Analysis using Chi-Square showed that all treatments caused pregnancy (100%). In summary,hCG injection at the time of artificial insemination and five days after artificial insemination in dairy cowsexperiencing repeat breeding can cause pregnancy.

Hypogonadism in the Prader-Willi syndrome from birth to adulthood: a 28-year experience in a single centre.

BackgroundHypogonadism is a key feature of Prader–Willi syndrome (PWS) but clear strategies for hormone replacement are lacking.ObjectiveTo evaluate the gonadal status and outcome in patients attending a Scottish PWS clinic from 1991 to 2019.MethodsIn 93 (35F:56M) patients, median follow-up 11.2 years, gonadal and pubertal status were assessed clinically. Pelvic ultrasound findings and basal/stimulated gonadotrophins were compared with age-matched controls.ResultsFemales:of 22 patients aged > 11, 9 had reached B4–5, while 5 were still at B2–3, and 6 remained prepubertal. Eight patients experienced menarche aged 9.8–21.4 years, none with a normal cycle. Uterine length and ovarian volumes were normal but uterine configuration remained immature, with low follicular counts. Gonadotrophins were unremarkable, serum oestradiol 129 (70–520) pmol/L. Only 5 patients received oestrogen replacement. Males:fifty-four (96%) patients were cryptorchid (9 unilateral). Weekly hCG injections resulted in unilateral/bilateral descent in 2/1 of 25 patients. Of 37 boys aged > 11, 14 (9 with failed/untreated bilateral cryptorchidism) failed to progress beyond G1, 15 arrested at G2–3 (testes 3–10 mL), and 8 reached G4–5. Gonadotrophins were unremarkable except in boys at G2–5 in whom FSH was elevated: 12.3/27.3 vs 3.25/6.26 U/L in controls (P < 0.001). In males aged > 13, testosterone was 3.1 (0.5–8.4) nmol/L. Androgen therapy, given from 13.5 to 29.2 years, was stopped in 4/24 patients owing to behavioural problems.ConclusionDespite invariable hypogonadism, few females and only half the males with PWS in this study received hormone replacement. Double-blind placebo-controlled crossover trials of sex steroids are required to address unproven behavioural concerns.

P–412 Low serum hCG levels adjusted for the hCG trigger dose in fresh cycles may be associated with a poor clinical pregnancy rate of FET cycles

Abstract Study question Is there any association between serum human chorionic gonadotropin (hCG) levels after trigger at previous fresh cycles and pregnancy outcomes of frozen-thawed embryo transfer (FET) cycles? Summary answer Low adjusted serum hCG level after hCG trigger at fresh cycles is negatively associated with clinical pregnancy rates (CPR) of hormone replacement treatment-FET (HRT-FET). What is known already: Literature showed that low serum hCG levels after the same dose of hCG trigger was associated with reduced pregnancy outcomes of the fresh cycles. However, the relationship between hCG levels after trigger at fresh cycles and pregnancy outcomes of FET cycles remains unknown. Study design, size, duration This matched retrospective study was conducted at a Reproductive Medicine Center between 2016 and 2018. Subjects performing HRT-FET cycles, whose previous fresh cycles used a bolus of hCG alone or a bolus of GnRHa combined with hCG for trigger were included. A total of 186 HRT-FET cycles with complete data was included for the final analysis. Participants/materials, setting, methods The study population was grouped into women with intramuscular injection of hCG prior to secretory transformation (hCG group, n = 93) and a comparison group (control group, n = 93) of women without hCG addition matched for patients’ age and duration of infertility. At the previous fresh cycles, serum hCG levels were measured 12 hours later after hCG trigger (defined as the “hCG+12 h” timepoint), and were adjusted for doses (defined as adjusted hCG levels). Main results and the role of chance: For patients achieving clinical pregnancy, the adjusted hCG level significantly increased (P &amp;lt; 0.05). Meanwhile, the ROC curve also showed a significantly predictive value of adjusted serum hCG levels at the “hCG+12 h” timepoint for CPR in HRT- FET cycles (AUC=0.626, 95%CI: 0.512–0.740) and the optimal hCG threshold proposed by ROC for CPR was 46.31 mIU/mL with sensitivity of 71.4% and specificity of 56.9%. For all patients, the CPR in hCG group was significantly higher than that in control group (61.3% vs. 44.1%). Furthermore, all cycles were then divided into four groups based on the injection of hCG prior to secretory transformation in HRT-FET cycles and this cut-off value of hCG levels at the “hCG+12 h” timepoint. For patients with adjusted hCG levels ≤46.31 mIU/mL, the CPR was significantly improved in hCG group compared with control group (61.1% vs. 29.3%). But for patients with adjusted hCG levels &amp;gt;46.31 mIU/mL, no statistically significant difference was observed between the hCG and control group (61.4% vs. 55.8%). Limitations, reasons for caution Although the results achieved statistically significant, the sample size was relatively small, which limits our ability to draw a definitive conclusion. The reason of the small sample size may be that to reduce the risk of OHSS, doctors would give preference to trigger with GnRH agonist in our center. Wider implications of the findings: Adjusted serum hCG levels might represent a potential factor to guide adequate support in the subsequent HRT-FET cycles. Meanwhile, for patients with low adjusted serum hCG levels, intramuscular hCG injection prior to secretory transformation may be a good compensation way to rescue pregnancy impair in the subsequent HRT-FET cycles. Trial registration number N/A.

Use of hCG hormone in the natural and artificial reproduction of Nile tilapia ( Oreochromis niloticus )

This study aimed to develop a protocol to synchronize final oocyte maturation and oocyte collection in both natural (first experiment) and artificial reproduction (second experiment) of Nile tilapia. In the first experiment, 80 young females (1 year old), 40 old females (3 years old) and 75 young males were used and splitted into three different groups. Each group bred once by natural mating during 7 days. Each female received an injection of hCG (3 IU/g of female) or saline solution (control) just before being stocked with males. In the second experiment, we evaluated different doses (0.5, 1.5, 2.5 and 3.5 IU/g female), number of injections (1 or 2) and administration route (intracoelomic or intramuscular) of hCG hormone in 40 females for artificial reproduction by stripping. The use of hCG (3 IU/g of female) was not effective for synchronization of natural reproduction in Nile tilapia, and the negative effect of age on reproductive performance was not reversed by its use. In the second assay, the use of a single injection with lower doses of hCG (0.5 and 1.5 IU/g of female) resulted in 100% of spawned females in artificial reproduction.

GnRHa implants and temperature modulate Lambari‐do‐rabo‐amarelo, Astyanax altiparanae (Characiformes: Characidae) induced reproduction out of the reproductive season

We evaluated the effect of gonadotropin in controlled-release implant together with temperature manipulation in the reproduction of Astyanax altiparanae (lambari) during the non-breeding season. Three groups of adult females were maintained at 20°C and three at 28°C. For each temperature, one group was treated with 4 μg GnRHa g−1, other with 8 μg GnRHa g−1 and other was sham implanted as a control during 4 weeks. Afterwards, 4 females/experimental group were induced to spawn with human chorionic gonadotropin, while 4 females/group were injected with saline. GnRHa implants increased fshβ and lhβ gene expression in females maintained at 28°C. The vtgA gene expression increased in the high GnRHa dose at 20°C, while this higher GnRHa dose associated with the higher temperature increased Gonadosomatic Index. The hCG injection increased the plasma Mature Induction Steroid levels and trigger ovulation, except in control at 28°C. The control females at 20°C spawned but with low reproductive performance, confirming that this is not a limiting temperature for reproduction. The administration of GnRHa as implant increased gonadotropin gene expression at 28°C, temperature at which GSI also increased. Qualitatively, the implants improved reproduction at 20°C, providing offspring out of the reproductive season, improving lambari production throughout the year.

PERKEMBANGAN EMBRIO PREIMPLANTASI SECARA IN VIVO SETELAH PEMBERIAN EKSTRAK ETHANOL AKAR ALANG-ALANG (IMPERATA CYLINDRICA L) PER ORAL

Cogon grass (Imperata cylindrica L) is known as a medicinal plant that is scattered almost worldwide. Despite its role that inhibits another plant’s growth, cogon grass possesses several benefits in health. This research has to identify the effect of short-term gavage ethanolic extract of cogon grass roots (CGG) to in-vivo mice preimplantation embryo development. A total of 60 female mice were divided into control and treatment groups, dosages at 90 and 115 mg/kg of body weight of CGG, orally gavage for 20 days. The superovulation of mice was done at the end of the CGG treatment by injecting 5 IU Pregnant Mare Serum Gonadotropin (PMSG) and after 48 hours, followed by 5 IU Human Chorionic Gonadotropin (hCG) injection and directly the mice were mated. The mating rate was checked by the appearance of the vaginal plug 12 hours after hCG injection. Mice were sacrificed, the oviducts and cornua of uteri were isolated to collect the oocytes and embryonic cells by flushing the oviducts and cornua uteri with Phosphate-buffered saline (PBS). The effects of CGG as an antifertility were evaluated by measuring the number of oocytes, fertilization, and in-vitro embryo development rates. The results showed significantly reduced about half of the mating rate in the 115 mg/kg BW group (p&lt;0.05) compared to control. However, the 90 mg/kg BW dose reduced 20% mating rate compared to control, and not significant (p&gt;0.05). In all treatment groups, only half oocytes fertilized. The cleavage and blastocyst rate in 115 mg/kg BW group were significantly lower compared to the control group (p&lt;0.05). In conclusion, oral gavage of cogon grass root ethanolic extract disrupts the mating process and development of in-vivo mice preimplantation embryo development.

Intrauterine injection of HCG before embryo transfer: a parallel, double-blind randomized trial

Research questionDoes intrauterine administration of HCG before embryo transfer improve live birth rate during IVF cycles? DesignA parallel, randomized controlled trial conducted between July 2018 and February 2020. Infertile women (n = 181) scheduled for fresh or vitrified–warmed embryo transfer after IVF carried out for any indication were randomized in a 1:1 ratio to receive either HCG (500 IU in 0.1 ml of tissue culture media) or culture media (0.1 ml of tissue culture media) via intrauterine injection 4 min before embryo transfer. In both groups, an intrauterine insemination catheter was used for administering the medication. Primary outcome was live birth, with ongoing pregnancy and clinical pregnancy as secondary outcomes. Analysis was based on intention-to-treat principle. ResultsBaseline and cycle characteristics were comparable between the two groups. In the control group, one woman with a confirmed clinical pregnancy was lost to follow-up. Live birth rates were 24% (22/90) in the HCG group versus 19% (17/90) in the control group (RR 1.29, 95% CI 0.74 to 2.27). Clinical pregnancy and ongoing pregnancy rates were 34% versus 26% (RR 1.31, 95% CI 0.84 to 2.04) and 24% versus 19% (RR 1.29, 95% CI 0.74 to 2.27) in the HCG and the control groups, respectively. ConclusionIntrauterine injection of HCG before embryo transfer did not improve live birth rates in women undergoing IVF. As the study was designed to detect a 20% difference between groups, a smaller, clinically important difference could not be ruled out. Treatment outcomes were lower than expected in the control group.

Vaginal stimulation enhances ovulation of queen ovaries treated using a combination of eCG and hCG.

Follicular changes throughout the oestrous phase have been poorly documented in queens because of the location and the small size of ovaries. We investigated follicular development in queens treated with a combination of equine chorionic gonadotropin (eCG) and human chorionic gonadotropin (hCG) and evaluated the effects of vaginal stimulation by a tomcat on ovulation induction. A hormonal treatment was administered using a simple crossover design. Four queens were administered 150 IU of eCG (day 1) and 250 IU of hCG on day 5 and 6. Half of the queens were mated with a vasectomised tomcat for 3 days after hCG injection. Ultrasound imaging of the ovaries clamped at a subcutaneous site was performed once a day from day 1 to 7, and on day 13, and the serum concentrations of oestradiol and progesterone were examined on day 1, 5, 7 and 13. The mean number of follicles gradually increased with the eCG treatment and decreased after hCG injection. The ovulation rate of follicles was significantly higher in the vaginal stimulation group (70.0%) than in the control group (42.6%). During the hormonal treatments, the serum concentration of oestradiol and progesterone did not differ between the two groups. Ultrasound imaging of the ovaries clamped at a subcutaneous site showed that eCG and hCG treatment promoted the follicular growth and corpus luteum formation, respectively. The combination of hCG injection with vaginal stimulation by a vasectomised tomcat enhanced the ovulation rate of follicles.