Clinical and Experimental Reproductive Medicine-CERM最新文献

Objective: This study aimed to compare the expression of implantation-related genes in trophectoderm cells between normally developing (day 5) and delayed (day 6) euploid blastocysts.

Methods: Trophectoderm biopsies were performed on blastocysts at either day 5 or day 6 of development. Preimplantation genetic testing for aneuploidy was conducted using next-generation sequencing. The expression levels of GAPDH, CXCR4, CXCL12, HSD3B1, HSD17B1, ITGAV, LAMA1, and MUC15 were measured from the remaining trophectoderm samples of euploid blastocysts using real-time quantitative polymerase chain reaction. Differences in gene expression levels between day 5 and day 6 euploid blastocysts were analyzed with the 2-ΔΔCt method. Pregnancy outcomes were assessed following single embryo transfer.

Results: Euploid blastocysts from day 5 (n=10) and day 6 (n=10) were included in the analysis. The expression level of CXCR4 was significantly lower in day 5 blastocysts compared to day 6 blastocysts (p<0.031). CXCL12, HSD17B1, ITGAV, and LAMA1 exhibited upregulated expression, while HSD3B1 and MUC15 showed downregulation in both day 5 and day 6 blastocysts, without significant differences between groups. Although gene expression did not differ significantly between pregnant and non-pregnant groups, early upregulation of CXCR4 was observed in the non-pregnant group following day 5 embryo transfer.

Conclusion: As the only gene associated with implantation, CXCR4 displayed a distinctive expression profile that increased during blastocyst development. Aberrant CXCR4 expression at specific stages of blastocyst development may influence pregnancy outcomes.

Objective: This study investigates the estrogenic activity of dibutyl phthalate (DBP) and its potential effects on endometrial receptivity and early embryo implantation. Given its widespread exposure and structural similarities to endocrine-disrupting chemicals, the potential for DBP to interfere with implantation-a key factor in fertility-is explored.

Methods: The optimal concentration of DBP was determined using the Cell Counting Kit-8 assay. Gene and protein expression related to attachment, estrogen receptor signaling, and inflammation were analyzed by reverse transcription quantitative polymerase chain reaction and western blotting. Immunocytochemistry was used to assess the nuclear translocation of estrogen receptor alpha (ERα), while attachment and outgrowth assays evaluated the effects of DBP on trophoblast behavior. Statistical analyses were performed using GraphPad Prism 5.01 (GraphPad Software) and SPSS ver. 18.0 (SPSS Inc.).

Results: DBP did not show cytotoxicity in Ishikawa cells at concentrations of 1, 10, and 100 μM. DBP treatment upregulated the expression of genes related to attachment, estrogen receptor signaling, and inflammation. Protein analysis showed an increase in inflammation-related proteins, and DBP enhanced ERα nuclear translocation. In trophoblast experiments, DBP-treated cells exhibited slightly lower attachment rates at early time points, but no significant differences were observed after 1 hour. DBP also reduced the outgrowth area of JEG-3 cells, with a significant decrease observed at 100 μM.

Conclusion: DBP exhibits estrogenic activity, disrupting implantation and invasion, and may pose risks to female reproductive health. These findings highlight the need for further investigation into the long-term effects of DBP exposure.

Objective: Lead acetate exposure induces male reproductive toxicity through oxidative stress and inflammation, impairing spermatogenesis and testosterone production. Umbelliferone (UMB), a coumarin derivative with antioxidant and anti-inflammatory properties, may counteract these adverse effects. This study evaluated the protective effects of UMB on lead acetate-induced testicular toxicity in male Wistar rats, with a focus on sperm parameters, antioxidant status, inflammatory markers, and testicular histology.

Methods: Thirty-two male Wistar rats were assigned to four groups (n=8 each): control (saline), lead (50 mg/kg lead acetate [intraperitoneal], lead+UMB (25 mg/kg), and lead+UMB (50 mg/kg). Treatments were administered daily for 21 days. Sperm parameters (count, motility, viability, morphology) were assessed, alongside measurements of antioxidant enzyme levels (superoxide dismutase, catalase, glutathione), malondialdehyde (MDA), serum testosterone, and mRNA expression of tumor necrosis factor-α, interleukin-6 (IL-6), transforming growth factor-β, IL-10, Bcl-2-associated X protein (Bax), and B-cell lymphoma-2 (Bcl-2). Testicular histology was evaluated using hematoxylin and eosin staining.

Results: Lead exposure significantly reduced sperm quality, antioxidant enzyme levels, testosterone, and Bcl-2 expression, while increasing MDA, pro-inflammatory cytokines, and Bax expression (p<0.05). UMB (25 and 50 mg/kg) markedly improved sperm parameters, restored antioxidant levels, reduced MDA and inflammatory markers, increased testosterone and Bcl-2, and decreased Bax expression (p<0.01). Histological analysis demonstrated that UMB preserved testicular architecture. No significant differences were observed between the two UMB doses (p>0.05).

Conclusion: UMB effectively mitigates lead-induced testicular toxicity by reducing oxidative stress, inflammation, and apoptosis, while improving sperm quality and testosterone levels. These findings suggest its potential as a therapeutic agent.

Objective: Cisplatin, a widely used chemotherapy agent, is known to induce testopathy and degeneration of the germinal epithelium (GE). Origanum vulgare L. (OV) leaf extract, due to its antioxidative properties, may alleviate such cellular damage. This experimental study was conducted to evaluate the protective and therapeutic effects of OV against cisplatin-induced testopathy.

Methods: Forty-eight male Naval Medical Research Institute mice were assigned to six groups. Testopathy was induced via intraperitoneal injection of cisplatin (single dose, 1 mg/kg on day 0). OV was administered as treatment (400 mg/kg orally, 5 days per week, for 5 weeks). Phytochemical screening of OV was also performed. After the experimental period, the animals were euthanized, and both blood serum and testicular samples were collected. Total body weight and total testicular weight were measured. Histopathological examination using hematoxylin and eosin staining (to assess the gonadosomatic index [GSI]) and immunohistochemical (IHC) detection of 3β-hydroxysteroid dehydrogenase (3β-HSD) protein were conducted. Expression levels of the p53 and B-cell lymphoma 2 (Bcl-2) genes, as well as serum testosterone levels, were evaluated. Statistical analysis was performed with SPSS ver.16, and significance was set at p<0.05.

Results: A phytochemical analysis of OV confirmed the presence of antioxidant compounds. Cisplatin administration resulted in significant detrimental alterations in testicular tissue (p<0.05). In animals receiving OV following cisplatin exposure, the GSI, testosterone levels, histological parameters, and total testicular weight improved toward physiological values (p<0.05). Additionally, IHC staining for 3β-HSD protein indicated regeneration of Leydig cells. Gene expression analysis showed down-regulation² of p53 and up-regulation of Bcl-2 (p<0.05).

Conclusion: OV administration, owing to its antioxidative characteristics, shows promise as a protective phytomedicine against cisplatin-induced testopathy. OV promotes GE proliferation, enhances testosterone secretion, and modulates the expression of apoptotic genes.

Objective: Lipopolysaccharide (LPS), derived from various infectious bacteria in the uterus, interferes with communication between embryonic trophoblasts and endometrial cells, thereby inhibiting successful embryo implantation. This study aimed to investigate the effects of LPS and the anti-inflammatory compound nicotinamide (NAM) on early embryo implantation processes, focusing on the adhesion and outgrowth between trophoblast spheroids and endometrial cells.

Methods: We used JAr mixed JEG-3 (JmJ) spheroids, prepared by combining JAr and JEG-3 cells in a 1:1 ratio. Following treatment with LPS with or without NAM, the attachment and outgrowth of JmJ spheroids on endometrial epithelial cells (ECC-1) were assessed. Additionally, changes in the gene expression of inflammatory cytokines (chemokine (C-X-C motif) ligand 1 [CXCL1], interleukin 8 [IL-8], and IL-33) and cell adhesion molecules (integrin alpha-V [ITGαV], integrin beta 3 [ITGβ3], and integrin beta 5 [ITGβ5]) in ECC-1 cells following LPS and/or NAM treatment were evaluated using quantitative reverse transcription polymerase chain reaction (qRT-PCR) and Western blot analysis.

Results: Decreased attachment rates and reduced outgrowth areas caused by LPS treatment were significantly restored by NAM. These restorative effects of NAM were associated with the modulation of inflammatory cytokines-specifically CXCL1 and IL-33, as shown by qRTPCR- and expression of the cell adhesion molecule ITGβ3, as indicated by Western blot analysis.

Conclusion: Our study confirmed that LPS-induced endometrial infection may inhibit embryo implantation. NAM treatment ameliorated the detrimental effects of LPS by modulating the expression of inflammatory cytokines and adhesion molecules. Further studies are needed to explore the potential use of NAM as an effective additive to improve embryo implantation rates in human in vitro fertilization-embryo transfer programs.

Objective: Despite advances in assisted reproductive technologies, predicting outcomes of in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) remains difficult. Hormonal status, oocyte maturity, and ovarian reserve contribute to treatment variability. This study examined correlations among demographic, endocrine, and embryological parameters in women undergoing IVF and ICSI whose partners had normal semen profiles, and evaluated the effect of follicle-stimulating hormone (FSH) levels on outcomes.

Methods: A retrospective analysis was performed on 488 women aged 18 to 45 years who underwent IVF and ICSI between 2022 and 2024. Data included age, body mass index (BMI), infertility duration, and levels of FSH, luteinizing hormone, anti-Müllerian hormone (AMH), thyroid-stimulating hormone, and fasting blood sugar. Embryological variables were oocyte yield, maturity stages (germinal vesicle, metaphase I, and metaphase II [MII]), and embryo count. Pearson correlations and the Kruskal-Wallis test were used to compare groups stratified by FSH (<10, 10-20, >20 mIU/mL).

Results: BMI and infertility duration showed weak correlations with embryological outcomes. AMH correlated positively with embryo count (r=0.29, p<0.01). MII oocytes correlated strongly with oocyte yield (r=0.90, p<0.01) and moderately with embryo count (r=0.46, p<0.01). Women with FSH <10 mIU/mL had significantly higher oocyte yield, MII oocyte numbers, and embryo counts than those with FSH ≥10 mIU/mL (p<0.001).

Conclusion: Lower FSH and higher AMH are associated with better oocyte maturity and embryo yield. These markers may support individualized stimulation strategies to improve IVF and ICSI outcomes.

Objective: Ovarian tissue cryopreservation is an essential fertility preservation technique. Two primary methods are used for ovarian tissue cryopreservation: slow freezing and vitrification. Recently, vitrification has been favored over slow freezing, and a closed system is recommended to prevent cross-contamination in liquid nitrogen. Follicular loss during freezing and thawing remains a major challenge. We investigated whether rapamycin, an inhibitor of the mechanistic target of rapamycin (mTOR) pathway, could mitigate primordial follicle loss during closed-system vitrification and thawing of mouse ovarian tissue.

Methods: Mouse ovaries were vitrified and thawed with or without 750 nanomolar rapamycin, then immediately analyzed or cultured for 5 days. Activation of the mTOR pathway was assessed using phosphorylated S6 kinase immunostaining, and follicle survival and development were evaluated by histological analysis.

Results: Closed-system vitrification did not induce apoptosis in primordial follicles. The median phosphorylated S6K-positive rate in primordial follicles was 7.1% in fresh controls, 87.9% in the rapamycin-free group, and 19.0% in the rapamycin-treated group (fresh-control vs. rapamycin-free and rapamycin-free vs. rapamycin-treated, both p<0.001). Rapamycin treatment suppressed this activation, resulting in significantly higher primordial follicle counts after culture (605 vs. 289 follicles per ovary, p<0.05) and a lower ratio of primary to primordial follicles, indicating reduced follicle activation.

Conclusion: These findings demonstrate that rapamycin preserves the primordial follicle pool by preventing follicle activation during cryopreservation and thawing. Incorporating rapamycin into closed-system vitrification protocols may improve ovarian tissue cryopreservation outcomes and enhance fertility preservation for patients with cancer.

In assisted reproductive technology, spermatozoa must be separated from seminal fluid to achieve optimal fertilization capacity. Conventional separation techniques frequently result in elevated reactive oxygen species production and iatrogenic injury due to repeated cell centrifugation. The aim of this systematic review and meta-analysis was to evaluate the effects of free centrifuge sorting (FCS) techniques on intracytoplasmic sperm injection (ICSI) outcomes. A comprehensive literature search was conducted using PubMed, Scopus, Web of Science, and Cochrane databases. The meta-analysis adhered to the Preferred Reporting Items for Systematic reviews and Meta-Analyses Protocols (PRISMA-P) guidelines. All eligible studies were selected using the population, intervention, comparison/comparator, outcomes, and study design (PICOS) methodology. The primary outcomes assessed were fertilization rate (FR), the high-quality embryo rate, implantation rate (IR), and clinical pregnancy rate (CPR). The study is registered in PROSPERO under registration number CRD42023415532. After screening 306 records for eligibility, three studies were ultimately included in the analysis. Our results demonstrate that following ICSI, a very brief period of abstinence significantly increased IR and CPR. However, no significant differences were observed for FR. The FCS technique yielded spermatozoa of superior biological quality following removal of seminal samples, and this purified sperm population improved reproductive outcomes in ICSI programs.

Objective: This study aimed to determine the optimal timing of cryopreservation and biopsy procedures in preimplantation genetic testing for aneuploidy (PGT-A) by comparing clinical outcomes between fresh embryo biopsy (fresh biopsy) and frozen-thawed embryo biopsy (frozen biopsy) procedures in high-risk patients.

Methods: This retrospective study included 844 patients undergoing 844 cycles conducted from August 2019 to December 2023. PGT-A was performed via trophectoderm biopsy using array comparative genomic hybridization and next-generation sequencing for comprehensive 24-chromosome screening. Patients were divided into two groups based on biopsy timing: fresh embryo biopsy (531 patients) and frozen- thawed embryo biopsy (313 patients).

Results: The clinical pregnancy rate was significantly higher in the fresh biopsy group compared to the frozen biopsy group (58.7% vs. 45.6%; odds ratio [OR], 1.695; 95% confidence interval [CI], 1.215 to 2.364; p=0.002). Furthermore, the fresh biopsy group showed higher implantation rates (45.6% vs. 32.1%; OR, 1.767; 95% CI, 1.274 to 2.451; p=0.002), ongoing pregnancy or live birth rates per cycle (48.0% vs. 35.8%; OR, 1.652; 95% CI, 1.177 to 2.319; p=0.004), and rates of good-quality blastocysts (57.1% vs. 32.1%, p<0.001) compared with the frozen biopsy group. Miscarriage rates did not differ significantly between the groups (18.2% vs. 21.4%; OR, 0.818; 95% CI, 0.457 to 1.465; p=0.501).

Conclusion: Fresh biopsy demonstrated superior clinical outcomes compared with frozen biopsy, likely due to better embryo quality. Both fresh and frozen biopsies remain viable options for PGT-A, with frozen biopsy serving as a practical alternative. Embryo quality and euploid status continue to be critical considerations for embryo transfer selection.

Objective: This study aimed to establish a quantitative and interpretable method for assessing oocyte quality by analyzing cytoplasmic morphology and intensity features using artificial intelligence.

Methods: A total of 695 oocyte images were collected from hormonally stimulated young and aged mice. The cytoplasmic region was manually annotated to exclude polar bodies, and radiomics analysis was performed to extract morphological and intensity-based features.

Results: Clustering with a Gaussian mixture model identified three distinct oocyte subtypes with unique cytoplasmic characteristics. Cluster 2, with the most spherical and compact oocytes, demonstrated the highest blastocyst formation rate (42.9%), followed by clusters 3 (35.3%) and 1 (20.4%). Cluster 2 oocytes also showed the highest mean intensity and lowest variability, suggesting uniform cytoplasmic structure. Notably, some aged oocytes in cluster 2 exhibited developmental potential comparable to that of young mice, indicating that cytoplasmic quality may be a more informative predictor than age alone.

Conclusion: These findings underscore the value of cytoplasmic features as objective indicators of developmental competence. This artificial intelligence-driven approach may improve embryo selection by providing a standardized, non-invasive method for evaluating oocytes, ultimately contributing to enhanced clinical outcomes in assisted reproductive technologies.