Two laser-assisted hatching methods of embryos in ART: a systematic review and meta-analysis.

BACKGROUND: Laser-assisted hatching (LAH) stands as the predominant technique for removing the zona pellucida (ZP) in embryos, primarily consisting of two methods: drilling laser-assisted hatching (D-LAH) and thinning laser-assisted hatching (T-LAH). Presently, both methods have limitations, and their comparative efficacy for embryo implantation and clinical pregnancy remains uncertain. AIM: Evaluate the impact of D-LAH and T-LAH on clinical pregnancy rates within assisted reproductive technology (ART). METHODS: We systematically searched electronic databases including PubMed, Web of Science, and Cochrane Library until July 20, 2022. This study encompassed observational studies and randomized controlled trials (RCTs). A 95% confidence interval (CI) was utilized for assessing the risk ratio (RR) of pregnancy outcomes. The level of heterogeneity was measured using I2 statistics, considering a value exceeding 50% as indicative of substantial heterogeneity. RESULTS: The meta-analysis scrutinized 9 studies involving 2405 clinical pregnancies from D-LAH and 2239 from T-LAH. Findings suggested no considerable variation in the clinical pregnancy rates between the two techniques (RR = 0.93, 95% CI: 0.79-1.10, I2 = 71%, P = 0.41). Subgroup analyses also revealed no substantial differences. However, D-LAH exhibited a notably higher occurrence of singleton pregnancies compared to T-LAH (RR = 2.28, 95% CI: 1.08-4.82, I2 = 89%, P = 0.03). There were no noteworthy distinctions observed in other secondary outcomes encompassing implantation rate, multiple pregnancies, ongoing pregnancy, miscarriage, premature birth, and live birth. CONCLUSION: Both the primary findings and subgroup analyses showed no marked variance in clinical pregnancy rates between D-LAH and T-LAH. Therefore, patients with varying conditions should select their preferred LAH technique after assessing their individual situation. However, due to the restricted number of studies involved, accurately gauging the influence of these laser techniques on clinical outcomes is challenging, necessitating further RCTs and high-quality studies to enhance the success rate of ART. TRIAL REGISTRATION: PROSPERO: CRD42022347066.

Predictors of vaginal delivery following balloon catheter for labor induction in women with one previous cesarean.

BACKGROUND: The aim of this study was to estimate predictors for vaginal birth following balloon catheter induction of labor (IOL) in women with one previous cesarean section (CS) and an unfavorable cervix. METHODS: This 4-year retrospective cohort study was conducted in Longhua District Central Hospital in Shenzhen China, between January 2015 and December 2018. Patients with one previous CS and a current singleton-term pregnancy who underwent balloon catheter cervical ripening and IOL were enrolled. Univariate analysis was used to identify predictive factors associated with vaginal birth after cesarean section (VBAC). Binary logistic regression was further used to identify which factors were independently associated with the outcome measure. The primary outcome was VBAC, which was a successful trial of labor after cesarean delivery (TOLAC) following IOL. RESULTS: A total of 69.57% (208/299) of the women who planned for IOL had VBAC. In the final binary logistic regression equation, lower fetal weight (< 4000 g) (odds ratio [OR]5.26; 95% confidence interval [CI] 2.09,13.27), lower body mass index (BMI,<30 kg/m2) (OR 2.27; CI 1.21, 4.26), Bishop score after cervical ripening > 6 (OR 1.94; CI 1.37, 2.76) remained independently associated with an increased chance of VBAC. CONCLUSIONS: The influencing factors of VBAC following IOL were fetal weight, BMI, and Bishop score after cervical ripening. Adequate individualized management and assessment of the IOL may help improve the VBAC rate.

Comparison of uterine, endometrial and subendometrial blood flows in predicting pregnancy outcomes between fresh and frozen-thawed embryo transfer after GnRH antagonist protocol: a retrospective cohort study.

This retrospective cohort study aimed to compare endometrial receptivity and pregnancy rate between fresh embryo transfer (ET) and frozen-thawed ET after gonadotrophin-releasing hormone (GnRH) antagonist protocol in normal ovarian responders. The patients were divided into two groups: the fresh ET group and the frozen-thawed ET group. Uterine artery resistance index (RI) and endometrial thickness were lower in the frozen-thawed ET group. The proportion of detectable endometrial-subendometrial flow was significantly higher in the frozen-thawed ET group. There was no significant difference in miscarriage rate between the two groups. Frozen-thawed ET group had a significantly higher CPR (56.0% vs. 48.1%), implantation rate (32.2% vs. 26.4%), and LBR (45.4% vs. 36.5%) than the fresh ET group. In GnRH antagonist protocol, elective frozen-thawed ET should be ideally taken, as this could improve embryo implantation rate, clinical pregnancy rate, and live birth rate, thus presenting an effective strategy to enhance the embryo utilization rate.IMPACT STATEMENTWhat is already known on this subject? The clinical pregnancy rate following fresh embryo transfer (ET) was lower than frozen-thawed ET after GnRH antagonist protocol. IVF success depends on embryo quality, embryo-endometrium interaction and endometrial receptivity. A good blood supply toward the endometrium is generally considered a requirement for implantation.What do the results of this study add? Uterine artery RI and endometrial thickness were significantly lower in the frozen-thawed ET group. The proportion of detectable endometrial-subendometrial flow was significantly higher in the frozen-thawed ET group. Frozen-thawed ET group had a significantly higher clinical pregnancy rate, implantation rate and live birth rate than the fresh ET group after GnRH antagonist protocol.What are the implications of these findings for clinical practice and/or further research? In GnRH antagonist protocol, elective frozen-thawed ET should be ideally taken, as this could improve embryo implantation rate, clinical pregnancy rate and live birth rate, thus presenting an effective strategy to enhance the embryo utilization rate.

[Relationship of the kallistatin gene expression with male spermatogenic dysfunction and its possible mechanism].

OBJECTIVE: To explore the role of the kallistatin gene in male spermatogenesis and its possible mechanism, and provide some new ideas for the clinical treatment of spermatogenic dysfunction. METHODS: We collected semen samples from the patients with oligospermia (OS) or non-obstructive azoospermia (NOAS) as well as testis tissue from testicular puncture. We detected the differential expression of kallistatin in the seminal plasma and the mRNA and protein expression levels of kallistatin, KLK1, B2R, Bcl-2, casepase-3, Bax and other molecules in the testis tissue, assessed the testicular spermatogenic function using Johnsen's scores, examined the interstitial fibrosis in the testis by Masson and Sirius staining, and analyzed the correlation of the expression level of kallistatin with spermatogenesis, apoptosis and fibrosis. RESULTS: Kallistatin was highly expressed in the seminal plasma and testis tissue. The expression of kallistatin was significantly decreased in the seminal plasma (P < 0.05) and so were those of kallistatin, KLK1 and B2R in the testis tissue of the NOAS and OS patients compared with those in the normal controls (P < 0.01), but no statistically significant difference was found between the expression levels of kallistatin and KLK1 within the same group (P > 0.05). The expression of Bcl-2 in the testis tissue was significantly lower (P < 0.01) and those of Bax and Casepase-3 dramatically higher in the OS and NOAS patients than in the normal males (P < 0.01). Cell apoptosis was negatively correlated with the expression of kallistatin. The results of Masson and Sirius staining showed that the fibrosis of the testis tissue and the ratio of type I/III collagen fibers were markedly increased in the OS and NOAS patients in comparison with the normal controls, even more significantly in the NOAS than in the OS group. CONCLUSION: Decreased expression of kallistatin may be related to spermatogenic dysfunction, and the kallistatin expression plays a regulatory role in the testicular spermatogenesis, probably by regulating cell apoptosis and fibrosis in the testis tissue, but the specific mechanism needs to be confirmed by further studies.

MIF-mediated NF-κB signaling pathway regulates the pathogenesis of polycystic ovary syndrome in rats.

Polycystic ovary syndrome (PCOS) resulting from abnormal glucose metabolism is a relatively common and complex endocrine disorder among women in their reproductive years, However, the pathogenesis of PCOS is still unclear. The purpose of this study is to investigate the macrophage migration inhibitory factor (MIF) involvement of the nuclear factor (NF)-κB in rats with PCOS. Results indicated that testosterone promoted the increase in the levels of MIF and luteinizing hormone (LH) but inhibited the increase in the level of follicular stimulating hormone (FSH). The MIF antibody could alleviate the process of PCOS to a certain extent. Testosterone promoted the expression of interleukin 1-beta (IL-1ß), interleukin 6 (IL-6), Inducible nitric oxide synthase (iNOS), and tumor necrosis factor alpha (TNF-α); the MIF antibody could reverse this effect. Testosterone could inhibit the expression of NF-κB protein whereas MIF antibody could promote the expression in the ovarian cytoplasm. Testosterone promoted the expression of NF-κB protein in the nucleus, this effect also could be reversed by the MIF antibody. Hyperandrogenism activated the NF-κB pathway. After using the MIF antibody, this effect was reversed. This finding suggested that hyperandrogenism activated the NF-κB pathway through MIF. In short, increased MIF levels activated the NF-κB pathway in ovaries, leading to inflammation and the increase in the levels of relevant inflammatory indicators, which might be one of the important factors in the pathogenesis of PCOS.

Interaction of BDE-47 with nuclear receptors (NRs) based on the cytotoxicity: In vitro investigation and molecular interaction.

Polybrominated diphenyl ethers (PBDEs) are endocrine-disrupting chemicals that possess neuroendocrine and reproductive toxicity to humans and disturb thyroid hormone homeostasis, neurobehavior, and development. The most predominant congener of PBDEs in humans and other organisms is 2,2',4,4'-tetrabromodiphenyl ether (BDE-47); however, the molecular mechanisms underlying its cytotoxicity remain largely unknown. Here, we evaluated the toxic effect and underlying mechanism of nuclear receptors (NRs) induced by BDE-47 in SK-N-SH human neuroblastoma cells. The CCK-8 cell viability assay showed that the proliferation of human SK-N-SH cells exposed to BDE-47 was significantly inhibited in time- and dose-dependent manners, and flow cytometry showed that cell cycle was arrested at the S phase after BDE-47 exposure. Moreover, compared with the control group, the expression of retinoic acid receptor alpha (RXRα), pregnane X receptor (PXR), thyroid hormone receptors (TRs), and peroxisome proliferator-activated receptors (PPARs) at the mRNA and protein levels was significantly increased, as determined by quantitative PCR and western blot analysis, demonstrating that BDE-47 activated the NRs in vitro. Moreover, BDE-47 could bind to all four NRs in the affinity order of PPARγ > PXR > TRß > RXRα under molecular dynamics. Because RXR is the promiscuous dimerization partner for a large number of NRs, ZDock was used to calculate its interaction with other three NRs. Taking the number of hydrogen bonds and ZDock scores into account, the rank of docking ability between RXRα and the NRs was PXR > TRß > PPARγ. Further analysis of the interaction between BDE-47 and dimerized-NRs, the affinity order was RXRα > TRß > PXR > PPARγ via Glide. The results of this study demonstrated that BDE-47 interfered the cross-talk among NRs, especially the promiscuous RXRα, which might be critical for the harmonized re-adjustment of cytotoxicity and biological regulation. Our findings provide a better understanding of the mechanisms underlying toxic effects and intermolecular interaction induced by BDE-47.

[Expression of the kallistatin gene in rat corpus cavernosum].

OBJECTIVE: To investigate the expression of the kallistatin gene in the corpus cavernosum and search for some new molecular targets for the regulation of penile erectile function and treatment of ED. METHODS: Using reverse transcriptase-quantitative polymerase chain reaction (RT-qPCR), Western blot and immunofluorescence staining, we detected the expression of kallistatin in the rat corpus cavernosum and compared it with that in the aorta. RESULTS: The results of RT-qPCR and Western blot revealed both mRNA and protein expressions of kallistatin in the rat corpus cavernosal tissue, with no statistically significant difference from those in the aorta (P > 0.05). Immunofluorescence staining showed that kallistatin was expressed in both endothelial and smooth muscle cells in the corpus cavernosum and localized in the cytoplasm, with no statistically significant difference from its expression in the aorta (P > 0.05) either. CONCLUSIONS: The kallistatin gene is highly expressed in the corpus cavernosum and localized in cavernosal endothelial and smooth muscle cells, suggestive of its involvement in the cellular function of cavernosal endothelial and smooth muscle cells and its participation in the regulation of penile erectile function.

Cellular metabolomics reveals glutamate and pyrimidine metabolism pathway alterations induced by BDE-47 in human neuroblastoma SK-N-SH cells.

Polybrominated diphenyl ethers (PBDEs) as potential neurotoxicants in environment may possess hazards to human health. Previous studies have reported that PBDEs exposure could induce oxidative stress and disturb mitochondrial functions in mammalian cells. However, the toxicological mechanism remains to be clarified. In this work, the neurotoxic effect and underlying mechanism of 2,2',4,4'-tetrabromodiphenyl ether (BDE-47) was investigated by using human neuroblastoma SK-N-SH cells as an effective model. A liquid chromatography-mass spectrometry (LC-MS)-based metabolomics approach combined with cell viability assay was applied to elucidate the metabolic perturbations and relevant toxicological pathways upon BDE-47 exposure. Our results shown that the SK-N-SH cell viability decreased in a dose-dependent manner after exposure to BDE-47 at 24 h within the concentration range of 5-250 µM, and an IC50 value of 88.8 µM was obtained. Based on the dose-response curve and cell morphological observation, the 5 and 10 µM BDE-47 doses (equal to IC5 and IC10, respectively) were used for metabolomics study to capture the sensitive metabolic response following BDE-47 exposure. After BDE-47 treatment, nine metabolites were identified as potential biomarkers, and the most disturbed metabolic pathways were mainly involved in alanine, aspartate and glutamate metabolism, glutathione metabolism, tyrosine and phenylalanine metabolism, and pyrimidine metabolism, which imply that metabolic changes related to neurotransmitters, oxidative stress, and nucleotide-mediated signal transduction systems were the sensitive pathways mostly influenced. Our findings reported here may provide potential neurotoxic effect biomarkers and prompt deep understanding of the molecular and metabolic mechanisms triggered by BDE-47 exposure.

[Effects of hexabromocyclododecanes on retinoid X receptor α, pregnane X receptor and peroxisome proliferator-activated receptor γ].

OBJECTIVE: To investigate the effect of hexabromocyclododecanes( HBCDs) on cell proliferation and the expression of the three important cell nuclear receptor of retinoic X receptor α( RXRα), peroxisome proliferator-activated receptor-γ( PPARγ), pregnane X receptor( PXR) and their interaction in Neuro-2a(N2a). METHODS: Neuro-2a cells were treated with different concentrations of diastereoisomers, of HBCDs which were α-HBCD, ß-HBCD, γ-HBCD, respectively, and cell toxicity was analyzed using the cell counting kit-8( CCK-8) assay. The impact of HBCDs on cell cycles of Neuro-2a were analyzed by flow cytometry analysis, and the expression levels in mRNA and protein for the three nuclearreceptors( RXRα, PPARγ, PXR andits target genes CYP3A11) were determined by RT-PCR and Western blot, respectively. The interaction between the receptors of RXRα, PXR, PPARγ was explored by immunoprecipitation. RESULTS: Cytotoxicity of ß-HBCD was the greatest among the three diastereoisomers, it was significantly greater than α-HBCD, however cytotoxicity of γ-HBCD for the Neuro-2a cells couldn 't be determined. Moreover α-HBCD, ß-HBCD induced significant cytotoxicity in a time-dose-response relationship to Neuro-2a cells( P < 0. 05), IC_(50) of α-HBCD, ß-HBCD to Neuro-2a cells were 60. 07 and 10. 52 µmol/L, respectively. α-, ß-HBCD blocked the cell cycle at G2/M phase. The expression levels in mRNA and protein of RXRα, PPARγ, PXR, CYP3A11 were significantly increased after cells exposure to α-HBCD and ß-HBCD 24 h. An interaction between RXRα, PPARγ and PXR in Neuro-2a cells existed no matter before and after exposure to HBCD. CONCLUSION: α-HBCD, ß-HBCD inhibit proliferation of Neuro-2a cells, cell cycle mainly was arrested at G2/M phase. α-HBCD, ß-HBCD could up-regulated the expression levels of RXRα, PPARγ, PXR. Meanwhile, the expression of CYP3A11 which is downstream gene of PXR also significantly increased( P < 0. 05). Interaction between RXRα, PPARγ and PXR exist whether or not exposure to α-, ß-HBCD. The molecular mechanisms of interaction between the receptors need further study.

Endometrial preparation protocol of the frozen-thawed embryo transfer in patients with polycystic ovary syndrome.

PURPOSE: To assess which is the optimal protocol in terms of endometrial preparation prior to frozen-thawed embryo transfer (FET) in women with polycystic ovarian syndrome (PCOS) and to explore the effect in stimulated cycle with the addition of vaginal 17-ß oestradiol. METHODS: Five hundred and seventy-six patients with PCOS were prepared for FET using artificial cycle induced with oestradiol and progesterone supplementation (n = 291) and stimulated cycle induced by human menopausal gonadotrophin (HMG) within or without the addition of vaginal 17-ß oestradiol (n = 285). Then the FET was performed in a receptive endometrium. RESULTS: Endometrial thickness was similar (9.03 ± 1.65 vs. 9.12 ± 1.58, P > 0.05) in artificial and stimulated cycle. The two protocols resulted in clinical pregnancy rate (41.0 % vs. 41.6 %, P > 0.05), ongoing pregnancy rate (36.6 % vs. 34.7 %, P > 0.05), live birth rate (30.0 % vs. 31.7 %, P > 0.05), which were not statistically different. Nevertheless, the cancelled cycle rate made a significant difference (2.2 % vs. 5.4 %, P < 0.05). There is no significant difference in the clinical pregnancy rate in HMG, HMG added with vaginal oestradiol and HMG switch to vaginal oestradiol group (42.6 %, 41.1 %, and 33.3 %, respectively). CONCLUSIONS: The mean endometrial thickness, clinical pregnancy rate, ongoing pregnancy rate, live birth rate and implantation rate were similar in artificial and stimulated cycle for endometrial preparation prior to FET in PCOS. It was fine to add vaginal 17-ß oestradiol to stimulated cycle when necessary. However, stimulated cycles had a significantly higher cancelled cycle rate. We should follow the principles of individualization, securitization and optimization in endometrial preparation of the FET in patients with PCOS.

Design strategies and biological applications of ß-galactosidase fluorescent sensor in ovarian cancer research and beyond.

Beta-galactosidase (ß-galactosidase), a lysosomal hydrolytic enzyme, plays a critical role in the catalytic hydrolysis of glycosidic bonds, leading to the conversion of lactose into galactose. This hydrolytic enzyme is used as a biomarker in various applications, including enzyme-linked immunosorbent assays (ELISAs), gene expression studies, tuberculosis classification, and in situ hybridization. ß-Galactosidase abnormalities are linked to various diseases, such as ganglioside deposition, primary ovarian cancer, and cell senescence. Thus, effective detection of ß-galactosidase activity may aid disease diagnoses and treatment. Activatable optical probes with high sensitivity, specificity, and spatiotemporal resolution imaging capabilities have become powerful tools for visualization and real time tracking in vivo in the past decade. This manuscript reviews the sensing mechanism, molecular design strategies, and advances of fluorescence probes in the biological application of ß-galactosidase, particularly in the field of ovarian cancer research. Current challenges in tracking ß-galactosidase and future directions are also discussed.

Different Cytotoxicity Induced by Hexabromocyclododecanes on Mouse Neuroblastoma N2a Cells via Oxidative Stress and Mitochondrial Apoptotic Pathway.

Hexabromocyclododecane (HBCD) is widely used in polystyrene foams, building materials, and electrical equipment as a brominated flame retardant (BFR) and persists in the environment and human body matrix. It has attracted increased attention since its neuroendocrine disorder effects have been observed in humans and animals. However, studies evaluating the neurotoxicity of HBCD diastereoisomers and the potential mechanisms involved are still limited. In this study, we compared the cytotoxicity induced by the three HBCD diastereoisomers (i.e., α-, ß-, and γ-HBCD) in N2a cells and further investigated the underlying molecular mechanism. Our results showed that HBCD diastereoisomers decreased cell viability in the order of ß-HBCD > α-HBCD > γ-HBCD. Moreover, α-HBCD and ß-HBCD exposure led to different degrees of cell cycle disruption and oxidative stress of N2a cells, implying that oxidative stress-mediated differential cytotoxicity of HBCD diastereoisomers. The expressions of caspases and Bcl-2 were differentially regulated by α-HBCD and ß-HBCD, suggesting that the mitochondrial apoptosis pathway may be critical in HBCDs-mediated N2a cell toxicity. Therefore, our studies provided novel evidence for the underlying mechanisms of the distinct cytotoxicity of HBCD diastereoisomers.

CAPN2 correlates with insulin resistance states in PCOS as evidenced by multi-dataset analysis.

OBJECTIVE: IR emerges as a feature in the pathophysiology of PCOS, precipitating ovulatory anomalies and endometrial dysfunctions that contribute to the infertility challenges characteristic of this condition. Despite its clinical significance, a consensus on the precise mechanisms by which IR exacerbates PCOS is still lacking. This study aims to harness bioinformatics tools to unearth key IR-associated genes in PCOS patients, providing a platform for future therapeutic research and potential intervention strategies. METHODS: We retrieved 4 datasets detailing PCOS from the GEO, and sourced IRGs from the MSigDB. We applied WGCNA to identify gene modules linked to insulin resistance, utilizing IR scores as a phenotypic marker. Gene refinement was executed through the LASSO, SVM, and Boruta feature selection algorithms. qPCR was carried out on selected samples to confirm findings. We predicted both miRNA and lncRNA targets using the ENCORI database, which facilitated the construction of a ceRNA network. Lastly, a drug-target network was derived from the CTD. RESULTS: Thirteen genes related to insulin resistance in PCOS were identified via WGCNA analysis. LASSO, SVM, and Boruta algorithms further isolated CAPN2 as a notably upregulated gene, corroborated by biological verification. The ceRNA network involving lncRNA XIST and hsa-miR-433-3p indicated a possible regulatory link with CAPN2, supported by ENCORI database. Drug prediction analysis uncovered seven pharmacological agents, most being significant regulators of the endocrine system, as potential candidates for addressing insulin resistance in PCOS. CONCLUSIONS: This study highlights the pivotal role of CAPN2 in insulin resistance within the context of PCOS, emphasizing its importance as both a critical biomarker and a potential therapeutic target. By identifying CAPN2, our research contributes to the expanding evidence surrounding the CAPN family, particularly CAPN10, in insulin resistance studies beyond PCOS. This work enriches our understanding of the mechanisms underlying insulin resistance, offering insights that bridge gaps in the current scientific landscape.

The effect and mechanism of low-molecular-weight heparin on the decidualization of stromal cells in early pregnancy.

OBJECTIVE: This study aimed to analyze the effect of low-molecular-weight heparin (LMWH) on the decidualization of stromal cells in early pregnancy and explore the effect of LMWH on pregnancy outcomes. METHODS: Recurrent spontaneous abortion (RSA) mouse model (CBA/J × DBA/2) and normal pregnant mouse model (CBA/J × BALB/c) were established. The female mice were checked for a mucus plug twice daily to identify a potential pregnancy. When a mucus plug was found, conception was considered to have occurred 12 h previously. The pregnant mice were divided randomly into a normal pregnancy control group, an RSA model group, and an RSA + LMWH experimental group (n = 10 mice in each group). Halfway through the 12th day of pregnancy, the embryonic loss of the mice was observed; a real-time quantitative polymerase chain reaction was used to detect the messenger ribonucleic acid (mRNA) expressions of prolactin (PRL) and insulin-like growth factor-binding protein 1 (IGFBP1) in the decidua of the mice. Additionally, the decidual tissues of patients with RSA and those of normal women in early pregnancy who required artificial abortion were collected and divided into an RSA group and a control group. Decidual stromal cells were isolated and cultured to compare cell proliferation between the two groups, and cellular migration and invasion were detected by membrane stromal cells. Western blotting was used to detect the protein expressions of proliferating cell nuclear antigen (PCNA), cyclin D1, matrix metalloproteinase- (MMP) 2, and MMP-7 in stromal cells treated with LMWH. RESULTS: Compared with the RSA group, LMWH significantly reduced the pregnancy loss rate in the RSA mice (p < 0.05). Compared with the RSA group, the LMWH + RSA group had significantly higher expression levels of PRL and IGFBP1 mRNA (p < 0.01). LMWH promoted the proliferation, migration, and invasion of human decidual stromal cells; compared with the control group, the expression levels of MMP-2, MMP-7, cyclin D1, and PCNA proteins in the decidual stromal cells of the LMWH group increased (p < 0.05). CONCLUSIONS: The use of LMWH can improve pregnancy outcomes by enhancing the proliferation and migration of stromal cells in early pregnancy and the decidualization of stromal cells.

Identification of biomarkers associated with macrophage infiltration in non-obstructive azoospermia using single-cell transcriptomic and microarray data.

Background: Non-obstructive azoospermia (NOA) is a common clinical cause of male infertility. Research suggests that macrophages are linked to testicular function; however, their involvement in NOA remains unknown. Methods: To evaluate the importance of macrophages infiltration in NOA and identify the macrophage-related biomarkers, the gene-expression microarray data GSE45885 and the single-cell transcriptomic data GSE149512 were utilized from the Gene Expression Omnibus (GEO). A single-sample gene set enrichment analysis (ssGSEA) was conducted to investigate immune cell proliferation. The Seurat package was used for the single-cell data analysis, and the limma package was used to identify the differentially expressed genes between the NOA and normal samples. Moreover, we conducted a weighted gene co-expression network analysis (WGCNA) to identify the macrophage-related key modules and genes, and conducted Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO) analyses for the functional exploration. To identify the macrophage-related biomarkers, we conducted least absolute shrinkage and selection operator (LASSO) and support vector machine-recursive feature elimination (SVM-RFE) analyses. Real-time quantitative polymerase chain reaction (RT-qPCR) was used to verify the marker genes present in NOA. Results: We confirmed that open reading frame 72 gene on chromosome 9 (C9orf72) [area under the curve (AUC) =0.861] and cartilage-associated protein (CRTAP) (AUC =0.917) were the hub genes of NOA, and the RT-qPCR analysis revealed the critical expression of both genes in NOA. Conclusions: Through the combination of tissue transcriptomic and single-cell RNA-sequencing analyses, we concluded that macrophage infiltration is significant in different subtypes of NOA, and we hypothesized that C9orf72 and CRTAP play critical roles in NOA due to their high expression in macrophages.

Generation of Human Endometrial Assembloids with a Luminal Epithelium using Air-Liquid Interface Culture Methods.

The endometrial lining of the uterus is essential for women's reproductive health and consists of several different types of epithelial and stromal cells. Although models such as gland-like structures (GLSs) and endometrial assembloids (EnAos) are successfully established, they lack an intact luminal epithelium, which makes it difficult to recapitulate endometrial receptivity. Here, a novel EnAo model (ALI-EnAo) is developed by combining endometrial epithelial cells (EnECs) and stromal cells (EnSCs) and using an improved matrix and air-liquid interface (ALI) culture method. ALI-EnAos exhibit intact EnSCs and glandular and luminal epithelia, which recapitulates human endometrium anatomy, cell composition, hormone-induced menstrual cycle changes, gene expression profiles, and dynamic ciliogenesis. The model suggests that EnSCs, together with the extracellular matrix and ALI culture conditions, contribute to EnAo phenotypes and characteristics reflective of the endometrial menstrual cycle. This enables to transcriptionally define endometrial cell subpopulations. It anticipates that ALI-EnAos will facilitate studies on embryo implantation, and endometrial growth, differentiation, and disease.

A quasi-one-dimensional bulk thermoelectrics with high performance near room temperature.

Pursuing efficient thermoelectricity from low-dimensional materials has been highly motivated since the seminal work of Hicks and Dresselhaus. In fact, many superior thermoelectric materials like Bi2Te3, Mg3Sb2/Mg3Bi2 and SnSe are quasi-two-dimensional (q2D), though the advantages of two-dimensionality appear to be diverse and sometimes controversial. Here, we report on a remarkably high thermoelectric performance in TlCu3Te2, which is quasi-one-dimensional (q1D) with a further reduced dimension. The thermoelectric figure of merit zT along its q1D axis amounts to 1.3 (1.5) at 300 (400) K, rivaling the best ever reported at these temperatures. The high thermoelectric performances benefit from, on one hand, large power factors derived from a center-hollowed, pancake-like Fermi pocket with q1D dispersion at the edge of a narrow band gap, and on the other hand, small lattice thermal conductivities caused by the large and anharmonic q1D lattice consisting of heavy, lone-pair-electron bearing (Tl+) and weakly-bonded (Cu+) ions. This compound represents the first bulk material with quasi-uniaxial thermoelectric transport of application level, offering a renewed opportunity to exploit reduced dimensionality for high-performance thermoelectricity.

The Comparison of Two Whole-Genome Amplification Approaches for Noninvasive Preimplantation Genetic Testing (ni-PGT) and the Application Scenario of ni-PGT during the Fresh Cycle.

Recently, noninvasive preimplantation genetic testing (ni-PGT) using degenerate oligonucleotide primer PCR (DOP-PCR) and multiple annealing and looping-based amplification cycle (MALBAC)-based whole-genome amplification (WGA) methods has demonstrated predictable results in embryo testing. However, a considerable heterogeneity of results has been reported in numerous studies on these two WGA methods. Our aim was to evaluate the current WGA method for ni-PGT while further clarifying the applicable scenarios of ni-PGT in the fresh cycle. A total of 173 embryos were tested with trophectoderm biopsy and ni-PGT. In the whole preimplantation genetic testing, the clinical concordance rates of the detection results of DOP-PCR and MALBAC with the corresponding trophectoderm biopsy results were 64.12% (84/131) and 68.99% (89/129), respectively (P = 0.405). However, in the detection of abnormal embryos, the detection efficiency of ni-PGT is significantly improved [MALBAC: 96.55% versus 68.99% (P < 0.001); and DOP-PCR: 89.09% versus 64.12% (P < 0.001)]. In addition, the diagnostic efficiency of ni-PGT in low-quality blastocysts was significantly higher than that in high-quality blastocysts [MALBAC: 95.24% versus 51.85% (P = 0.001); and DOP-PCR: 91.30% versus 48.15% (P = 0.001)]. These results contribute to further understanding ni-PGT and to clarifying its application scenario in the fresh cycle.

Progress of Single-Cell RNA Sequencing Technology in Myocardial Infarction Research.

After myocardial infarction, the heart enters a remodeling and repair phase that involves myocardial cell damage, inflammatory response, fibroblast activation, and, ultimately, angiogenesis. In this process, the proportions and functions of cardiomyocytes, immune cells, fibroblasts, endothelial cells, and other cells change. Identification of the potential differences in gene expression among cell types and/or transcriptome heterogeneity among cells of the same type greatly contribute to understanding the cellular changes that occur in heart and disease conditions. Recent advent of the single-cell transcriptome sequencing technology has facilitated the exploration of single cell diversity as well as comprehensive elucidation of the natural history and molecular mechanisms of myocardial infarction. In this manner, novel putative therapeutic targets for myocardial infarction treatment may be detected and clinically applied.

Single-cell analysis of embryoids reveals lineage diversification roadmaps of early human development.

Despite its clinical and fundamental importance, our understanding of early human development remains limited. Stem cell-derived, embryo-like structures (or embryoids) allowing studies of early development without using natural embryos can potentially help fill the knowledge gap of human development. Herein, transcriptome at the single-cell level of a human embryoid model was profiled at different time points. Molecular maps of lineage diversifications from the pluripotent human epiblast toward the amniotic ectoderm, primitive streak/mesoderm, and primordial germ cells were constructed and compared with in vivo primate data. The comparative transcriptome analyses reveal a critical role of NODAL signaling in human mesoderm and primordial germ cell specification, which is further functionally validated. Through comparative transcriptome analyses and validations with human blastocysts and in vitro cultured cynomolgus embryos, we further proposed stringent criteria for distinguishing between human blastocyst trophectoderm and early amniotic ectoderm cells.