International Journal of Fertility & Sterility最新文献

Background: mtDNA4977 deletion, linked to impaired mitochondrial function, is reportedly elevated in the spermatozoa of infertile males. We hypothesized that increased deletion load could lead to reduced ATP and S-adenosylmethionine (SAM) levels, thereby disrupting the balance between DNA methylation and demethylation. This study aimed to investigate the relationship between mtDNA4977 deletion, ATP levels, SAM concentration, and global DNA methylation in sperm samples of infertile men.

Materials and methods: In this experimental study, semen samples were collected from 22 men with oligoasthenoteratozoospermia (OAT) and 22 controls. Sperm analysis was performed according to the World Health Organization criteria. Spermatozoa were isolated, and mtDNA4977 deletion was assessed using quantitative polymerase chain reaction (PCR). ATP levels were measured by bioluminescence, SAM levels were measured by ELISA, and global DNA methylation was measured by 5-methylcytosine (5-mC) quantification. Data were analyzed using the Mann-Whitney U test. Correlation values were calculated using Spearman's correlation.

Results: mtDNA4977 deletion was detected in all samples. The OAT group exhibiting a significantly higher deletion load comparing the control group (P<0.001). Surprisingly, ATP levels were significantly higher in the OAT group (P<0.001). Similarly, SAM levels were significantly elevated in the OAT group compared to the control group (P<0.001). No significant difference in global DNA methylation was observed between the two groups (P=0.052). Furthermore, Spearman's correlation analysis revealed no significant association between deletion load, ATP, SAM, and global methylation levels in men with OAT.

Conclusion: While we could not definitively determine the exact mechanism underlying the elevated ATP levels, we propose potential explanations, such as an increased mitochondrial copy number or the accumulation of unconsumed ATP due to motility defects. A possible reason for the lack of significant methylation differences may be the limitation of examining global methylation without assessing gene-specific methylation profiles in spermatozoa. Further research is needed to explore these findings and their implications for male infertility.

Background: The success rate of in vitro maturation (IVM) for human oocytes is clinically significant, prompting a focus on optimizing IVM media culture. While various factors have been incorporated to improve outcomes, the role of Ghrelin hormone, despite its multifunctional nature, remains poorly investigated. This study aimed to determine the most effective concentration of the growth hormone releasing protein-6 (GHRP-6), Ghrelin hormone agonist, in the culture medium.

Materials and methods: In this experimental study, a total of 240 human germinal vesicle (GV) oocytes were collected and cultured in varying concentrations of GHRP-6. Maturation rates were assessed during two days of culture, and compared against a blastocyst media (single-step culture) as control group and another IVM media, human tubal fluid (HTF) 10%, as the sham group. Additionally, the expression levels of two genes associated with nuclear and cytoplasmic maturation were compared in the number of 164 GV oocytes randomly cultured in the most effective concentration of GHRP-6, control and sham groups for 24 hours, using real-time polymerase chain reaction (PCR).

Results: The optimal concentration of GHRP-6 for the IVM procedure was determined to be 75 ng/ml, resulting in a maturation rate of 70% on the first day and 80% on the second day. These results surpassed those of other culture media on both days. Real-time PCR data indicated that, despite the early appearance of the first polar body (PB1) on 24h of culture, the Ghrelin agonist did not elevate the expression levels of CENP-E or LINGO2, genes associated with meiotic progression and membrane proteins, respectively.

Conclusion: In summary, while GHRP-6 showed potential in promoting nucleonic maturation by significantly inducing the appearance of PB1 between GVs within 24 hours, it did not exhibit statistically significant improvements in cytoplasmic maturation in metaphase 2 oocytes (MII) during this timeframe.

Background: Recurrent pregnancy loss (RPL) is a condition in which a woman experiences two or more consecutive miscarriages before reaching 20 weeks of gestation. The present study was designed to explore the potential link between the interleukin (IL)-33 and RPL. To achieve this goal, we conducted a comprehensive search across the ISI, PubMed, Embase, and Scopus databases.

Materials and methods: We reviewed studies that examined the relationship between specific IL-33 polymorphisms and RPL using the appropriate keywords. The comparator in our study consisted of a control group that included women without RPL. This group was used to assess the differences in genetic polymorphisms and IL-33 serum levels compared to women who experienced RPL. We analyzed the collected data using a random-effects model in STATA (version 14). Five case-control studies were eligible, yielding a total sample size of 1,831 participants (891 cases and 940 controls).

Results: The results revealed a significant association between the GG genotype of IL-33 polymorphisms rs16924159 and rs1929992 [odds ratio (OR)=0.72, 95% confidence intervals (CI)=0.51-1.02, I²=58.2, P=0.068] and RPL. The presence of the GG genotype of IL-33 polymorphisms rs16924159 and rs1929992 in women was associated with a 0.72-fold increased risk of RPL. Additionally, no statistically significant correlation was noted between IL-33 GA (OR=1.19, 95% CI=75-1.88, I²=80.7, P=0.001) and AA (OR=1.02, 95% CI=0.81-1.29, I²=71.8, P=0.029) genotypes and RPL. Analysis of IL-33 serum levels indicated a significant association with RPL (OR=-0.45, 95% CI=-0.67-(- 0.23), I²=57.3, P=0.128).

Conclusion: Our findings demonstrate an association between the GG genotype of IL-33 polymorphisms and an increased risk of RPL. Moreover, higher serum levels of IL-33 are likely protective against RPL.

Background: Myomectomy is commonly performed on women diagnosed with symptomatic uterine leiomyoma(s). This study aimed to evaluate the effects of myomectomy on the serum anti-Müllerian hormone (AMH) levels in women with uterine myoma.

Materials and methods: In this prospective quasi-experimental study, 93 patients with uterine leiomyoma aged 18- 45 years were enrolled and underwent open and laparoscopic myomectomy. The participants' baseline characteristics were recorded. The level of AMH was measured and recorded before and six months after the surgery for each patient. The size, number, and type of myoma, the duration of surgery, the volume of bleeding during surgery, the need for blood transfusion, and postsurgical complications were investigated at 6-month intervals after the surgery. Data were analyzed by SPSS version 26.

Results: The AMH level decreased significantly after the surgery compared to before the surgery in both groups of laparotomic and laparoscopic myomectomy patients (P<0.001). The rate of AMH drop was lower in the laparoscopy group than in the laparotomy group (P<0.001). Among the studied variables, changes in AMH level showed a direct and significant correlation with myoma size and type. Postoperative pain, fever, and surgical site infection (SSI) were the most frequent postsurgical complications. In post-surgical period, fever rate was 12.3% in the laparotomy group, and 6.1% in the laparoscopy group, and pain (measured by visual analogue scale) was higher in the laparotomy group compared to the laparoscopy group (21 vs. 7.3%). SSI rate was 0.9% in the laparoscopy group compared to 6.3% in the laparotomy group. The size of the myoma had no significant effect on the occurrence of these complications.

Conclusion: Myomectomy may lead to a significant decrease in AMH levels in women with uterine leiomyoma undergoing both open and laparoscopic myomectomies, and the size and type of myoma significantly affects the changes in the hormone.

Background: Idiopathic Male Infertility is a widespread problem affecting approximately 10 to 15 percent of men of reproductive age. Several genes have been studied related to idiopathic male infertility. Thereafter, among the family of non-coding RNAs, microRNAs are shown to play a very important role in the regulation of a subset of genes. It has been documented that infertile men have differentially expressed miRNA. The aim of this study is to evaluate the impact of two genetic variants, miR-222 rs2858060 and miR-146a rs2910164 on idiopathic male infertility.

Materials and methods: In this case-control study, Blood samples were taken from 201 men with idiopathic infertility and 201 men in a healthy state for this case-control investigation. Genotype determination of desired polymorphisms was done using the Tetra-ARMS PCR technique and data was analyzed by SPSS software version 20. Significant associations were determined at an alpha level of 0.05.

Results: The study found a significant association between the miR-222 rs2858060 GG genotype and idiopathic male infertility in the recessive genetic model [odds ratio (OR)=2.02, 95% confidence interval (CI)=1.25-3.24, P=0.004]. After adjusting for age, body mass index (BMI), smoking, and alcohol consumption, the association between the genotypes and the disease remained significant. No significant association was observed between miR-146a rs2910164 genotypes and male infertility in all genetic models (P>0.05). Moreover, haplotype analysis showed that the CC (rs2858060/ rs2910164) haplotype is associated with the decreased risk of idiopathic male infertility (OR=0.728, 95% CI =0.54-0.98, P=0.035).

Conclusion: miR-222 rs2858060 and miR-146a rs2910164 polymorphisms influence the risk of male infertility in Iranian population. This is the first report to examine the role of miR-222 rs2858060 and miR-146a rs2910164 in male infertility, further studies involving different ethnicity and larger sample sizes are needed to confirm our findings.

Background: Cryopreservation of immature testicular tissue is a suitable method for spermatogonial stem cell (SSC) preservation in prepubertal boys, who are at risk of infertility due to cancer treatments. Viable spermatozoa can be obtained by transplantation or in vitro culture of cryopreserved testicular tissue. Optimizing the culture conditions is essential for reducing tissue damage caused by oxidative stress produced during cryopreservation and culture. Our objective was to improve the culture conditions of vitrified immature mouse testicular tissue by using N-acetylcysteine (NAC) antioxidant.

Materials and methods: In this experimental study, testicular tissues of 6-day-old immature NMRI mice were isolated, vitrified, and distributed into three groups: control, culture I (cultured without NAC), and culture II (cultured in the presence of 125 mM NAC). After seven days of culture, histological analysis, cell viability, apoptotic-related gene expression, promyelocytic leukaemia zinc finger (Plzf) gene expression, and Caspase-3 protein expression were assessed. Moreover, the malondialdehyde (MDA) level was measured in the culture media.

Results: Tissue integrity and higher viability level were observed in the culture II group compared to the other two groups. Furthermore, the Bax/Bcl-2 ratio and MDA level were decreased significantly in the culture ӀӀ group, whereas Caspase-3 and Plzf gene expression were significantly increased.

Conclusion: Our data revealed that the presence of 125 mM NAC improves the developmental process of vitrifiedwarmed immature mouse testicular fragments during in vitro culture, thus it may have potential implications for in vitro culturing of human prepubertal testicular tissues.

 Background: We aimed to examine the therapeutic efficacy of exosome-enriched conditioned media (CM), known for its high concentration of extracellular vesicles (EVs), in comparision with mesenchymal stromal/stem cells (MSCs) in treating non-obstructive azoospermia.

Materials and methods: In this experimental study, we used adipose tissue-derived MSCs (AT-MSCs), bone marrowderived MSCs (BM-MSCs), and BMCM containing EVs to treat busulfan-induced azoospermia in animal models. The study included thirty adult male Balb/C mice and two female enhanced green fluorescent protein-positive (eGFP+/+) Balb/C mice for experimental groups and stem cell culturing. Groups consisted of an intact control, an azoospermia group, an AT-MSC therapy group, a BM-MSC therapy group, a BMCM therapy group, and a spontaneous healing group. Testes were removed from all mice, and histomorphometry and flow cytometry analyses were performed 60 days post-treatment. Additionally, protein structure extraction, protein-protein docking analysis, and data visualization were conducted.

Results: Histomorphometry and flow cytometry showed that most seminiferous tubules in the therapy groups exhibited normal morphology and restored spermatogenesis, unlike the azoospermia group. In silico protein docking analysis revealed that exosome factors in BM-MSCs positively impacted spermatogenesis. The BM-MSC and BMCM therapy groups showed more favorable outcomes compared to other groups. Key exosome factors like Basigin, E3 ubiquitinprotein ligase (UBR2), transforming growth factor beta (TGF-β), hepatocyte growth factor (HGF), and programmed death-ligand 1 (PD-L1) interacted with receptors critical to this process.

Conclusion: Our findings indicate that both BMCM enriched with EVs and the administration of AT-MSCs and BM-MSCs effectively induced spermatogenesis in mice with busulfan-induced azoospermia. Specifically, BM-MSC therapy exhibited superior outcomes compared to AT-MSCs and BMCM alone. This study highlights the potential of EV-based therapies, particularly BMCM, as a promising strategy for treating non-obstructive azoospermia. Furthermore, the interaction of key exosome factors with critical receptors enhances our understanding of the underlying molecular mechanisms involved in restoring reproductive function in testes.

The hyperandrogenic non-classic congenital adrenal hyperplasia (NCAH) is an autosomal recessive disorder that can lead to a decrease in reproductive function in women due to alterations in hormone levels. Couples in which the woman is affected must investigate the occurrence of the mutation in the partner to assess the possibility of passing on the condition on their offspring. In such cases, genetic counseling plays a crucial role in educating the patient about the risk of having a child with the same disorder. Additionally, assisted reproduction techniques (ART), including preimplantation genetic testing for monogenic diseases (PGT-M), are important to increase the chances of the couple conceiving a healthy child. In this case report, we describe a multidisciplinary approach aimed at achieving two infants who appeared unaffected by the classic form of the condition. The couple carried variants in CYP21A2 gene, and the woman, affected by NCAH, presented a novel variant.

Background: Endometriosis is among the leading causes of morbidity in the female population worldwide. Currently, the definite diagnosis of endometriosis depends on laparoscopy as the gold-standard method. Potential biomarkers, such as inflammatory biomarkers, cancer antigens, and hormones, offer non-invasive alternatives. This study was designed to investigate the utility of some hematological markers, including white blood cell (WBC) count, neutrophil to lymphocyte ratio (NLR), platelet count, cancer antigen 19-9 (CA 19-9), and 125 (CA-125), carcinoembryonic antigen (CEA), and anti Müllerian hormone (AMH), as non-invasive methods for endometriosis diagnosis.

Materials and methods: In this retrospective study, which was performed on 346 females, the case group consisted of 230 endometriosis patients. The data of 116 patients with benign tumors or other benign conditions such as Müllerian anomalies, were used as the control group. Receiver-operating characteristic (ROC) analysis was implemented to calculate specificities and sensitivities of CA-125, CA 19-9, CEA, NLR, WBC count, platelet (PLT) count, and AMH.

Results: Significantly higher mean values were observed for CA-125, CA 19-9, WBC count, and NLR in the case group compared with the control group (P<0.001). The combination of NLR and CA-125 demonstrated the highest diagnostic performance with an area under the curve (AUC) of 0.903. However, the AUC for CA-125 (0.896) was lower and the value of 12.7 IU/mL was the most appropriate cut-off point (sensitivity=93.9%, specificity=60.9%). The cut-off value of 35 for CA-125 was also evaluated (sensitivity=61.4%, specificity=98.3%). The AUC for NLR was 0.699 and the best cut-off point was 1.5 (sensitivity=83.4%, specificity=52.6%).

Conclusion: Combined measurement of CA-125 and NLR showed the highest performance in the diagnosis of endometriosis and can be considered as a diagnostic marker. However, it is necessary to conduct more research to evaluate the applicability of these biomarkers.

Background: Cervical, breast, and ovarian cancers exhibit significant incidence and fatality rates, necessitating diverse approaches for effective cancer cell eradication while preserving normal cells. The aim of this study is to explore the in vitro apoptosis-inducing properties of hydroalcoholic extracts from Cyperus rotundus' (C. rotundus) leaf using human gynecological cancer cell lines.

Materials and methods: In this experimental study, Hydroalcoholic extracts were prepared from the leaf of C. rotundus using ethanol, methanol, and ethyl acetate. These extracts were applied to MCF-7, HeLa, OVCAR-3, and Vero cell lines at concentrations of 3.125, 6.25, 12.5, and 25 g/ml. MTT test, assessing inhibition of proliferation at 50% (IC₅₀), was employed to evaluate each extract's ability to inhibit cell proliferation. Subsequently, apoptosis-related gene and protein regulation were examined using reverse transcription polymerase chain reaction (RT-PCR) and Western blot analysis.

Results: Gas chromatography-mass spectrometry (GC-MS) analysis revealed that the methanolic extract contained hexadecanoic acid and dodecanoic acid. The ethanolic extract was found to have norspermidine and desulphosinigrin. Additionally, the ethyl acetate extract included vitamin E, 1-heptatriacotanol, lupeol, betulin, stigmasterol, and stearic acid. The HeLa treatment group with 6.25 μg/ml of ethyl acetate extract, MCF-7, and OVCAR-3 cells with 3.125 μg/ ml of methanol extract treatment group exhibited the most significant growth inhibition in the MTT assay. Further analysis of these treatment groups revealed that the transcription and translation of BAX, Caspase-3, Caspase-8, and Caspase-9 increased overall, whereas Bcl-2 decreased in all cell lines.

Conclusion: Hydroalcoholic extracts from C. rotundus' leaf may enhance the apoptosis of cancer cells by modulating the transcription, translation, and post-translation of proteins, with minimal impact on the growth and survival of non-cancerous cells.