Infertility is a common reproductive disorder affecting millions of couples worldwide. It is estimated that male factors account for about 30%-50% of infertility cases, and some studies have found that the concentration of male sperm gradually decreases over time, a trend that suggests the importance of male fertility. Many factors contribute to the decline of male fertility, among which environmental factors have received widespread attention. After reaching adulthood, spermatogonial stem cells will continue to produce sperm, but these cells exist outside the blood testicular barrier, which makes them highly sensitive to environmental conditions such as air pollution, tobacco smoke, radiation, and heavy metals. It is reported that exposure to these adverse environmental factors not only causes oxidative stress and DNA damage to germ cells, but also leads to abnormal epigenetic modification of sperm DNA, thereby causing a series of diseases. This article reviewed the abnormal methylation changes in DNA associated with exposure to environmental pollutants during spermatogenesis and how these changes affect the quantity, quality, and function of spermatozoa.

PURPOSE: To investigate the regulatory role of nerve injury-induced protein 1 (NINJ1) in the anti-inflammatory function of human umbilical cord mesenchymal stem cells (hUC-MSCs) co-stimulated by interferon-gamma (IFN-γ) and tumor necrosis factor-alpha (TNF-α). METHODS: hUC-MSCs were expanded in vitro using standard protocols, with stem cell characteristics confirmed by flow cytometry and multilineage differentiation assays. The immunomodulatory properties and cellular activity of cytokine-co-pretreated hUC-MSCs were systematically evaluated via quantitative reverse transcription RT-qPCR, lymphocyte proliferation suppression assays, and Cell Counting Kit-8 viability tests. Transcriptome sequencing, Western blotting and small interfering RNA interference were integrated to analyze the regulatory mechanisms of NINJ1 expression. Functional roles of NINJ1 in pretreated hUC-MSCs were elucidated through gene silencing combined with lactate dehydrogenase release assays, Annexin V/Propidium Iodide apoptosis analysis, macrophage co-culture models, and cytokine Enzyme-Linked Immunosorbent Assay. Therapeutic efficacy was validated in a cecal ligation and puncture-induced septic mouse model: 80 mice were randomly allocated into 4 experimental groups (n=20/group): sham group (laparotomy without cecal ligation); phosphate-buffered saline-treated group (cecal ligation and puncture (CLP) + 0.1 mL phosphate-buffered saline); hUC-MSCs (small interfering RNA (siRNA)-interferon-gamma and tumor necrosis factor-alpha co-stimulation (IT))-treated group (CLP + hUC-MSCs transfected with scrambled siRNA); and hUC-MSCs (siNINJ1-IT)-treated group (CLP + hUC-MSCs with NINJ1-targeting siRNA). RESULTS: hUC-MSCs demonstrated compliance with International Society for Cellular Therapy criteria, confirming their stem cell identity. IFN-γ/TNF-α co-pretreatment enhanced the immunosuppressive capacity of hUC-MSCs, accompanied by the reduction of cellular viability, while concurrently upregulating pro-inflammatory cytokines such as interleukin-6 and interleukin-1β. This co-stimulation significantly elevated NINJ1 expression in hUC-MSCs, whereas genetic silencing of NINJ1 effectively suppressed pro-inflammatory cytokine production and attenuated damage-associated molecular patterns release through inhibition of programmed plasma membrane rupture. Furthermore, the NINJ1 interference potentiated the ability of cytokine-pretreated hUC-MSCs to suppress LPS-induced pro-inflammatory responses in RAW264.7 macrophages. In cecal ligation and puncture-induced sepsis model, NINJ1-silenced hUC-MSCs exhibited enhanced therapeutic efficacy, manifested by reduced systemic inflammation and multi-organ damage. CONCLUSION Our findings shed new light on the immunomodulatory functions of cytokine-primed MSCs, offering groundbreaking insights for developing MSC-based therapies against inflammatory diseases via interfering the expression of NINJ1.
Mesenchymal Stem Cells/drug effects* ; Animals ; Interferon-gamma/pharmacology* ; Tumor Necrosis Factor-alpha/pharmacology* ; Humans ; Mice ; Umbilical Cord/cytology* ; Cells, Cultured ; Apoptosis ; Male

Alcohol exposure, as a widespread environmental factor, is highly toxic and teratogenic. Embryonic stem cells (ESCs) are pluripotent and key to development, and their gene expression is tightly regulated, allowing the cells to differentiate without self-renewal. Numerous studies showed that alcohol is an important factor affecting the differentiation of ESCs. In this paper, we systematically summarized four major molecular mechanisms underlying alcohol associated differentiation of ESCs: (1) inhibition of the Wnt signaling pathway; (2) restriction of the mitogen-activated protein kinase/extracellular signal-regulated kinase (MAPK/ERK) pathway; (3) alteration of the expression of pluripotent transcription factors; and (4) activation of the nuclear transcriptional program. Through the above mechanisms, alcohol induces aberrant expression of differentiation-related genes and alters the direction of cellular differentiation towards specific lineages, thereby affecting normal embryonic development. Based on the studies on ESCs modeling and other in vitro and in vivo differentiation experiments, the molecular basis of how alcohol affects differentiation by interfering with signaling networks and transcriptional regulation was elucidated, and the results of current research in this field were also summarized, which is crucial for understanding alcohol-mediated toxic effects.

Objective:Analyzing the influence of morphological evaluation parameters of blastocysts, including days of blastocyst development [day 5 (D5) and day 6 (D6)], degree of blastocyst expansion (4, 5, 6), inner cell mass and trophectoderm grade, on the occurrence of chromosomal karyotype abnormalities of chorionic villi in missed abortion after in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) treatment and fresh/frozen-thawed single blastocyst transfer. Methods:The clinical data of patients with missed abortion after IVF/ICSI treatment and fresh/frozen-thawed single blastocyst transfer from February 2015 to February 2023 in the Reproductive Center of the Third Affiliated Hospital of Zhengzhou University were included. Using a case-control study, the data were divided into two groups according to the detection results of chromosomal copy number variations (CNVs) in chorionic villi of missed abortion abnormal karyotype group ( n=139) and normal karyotype group ( n=82). The baseline data between the two groups were compared. Univariate logistic regression was used to investigate the effect of blastocyst morphological rating parameters on the occurrence of chromosomal karyotype abnormalities of chorionic villi in aborted tissues, and multivariate logistic regression was also used to adjust confounding factors. Results:Male age [(34.12±6.49) years], sperm morphology rate [5.00 (4.00,6.00)%] and female age [33.00 (30.00, 37.00) years] in abnormal karyotype group were higher than those in the normal karyotype group [(32.38±4.69) years, 4.00 (2.00,5.00)% and 31.50 (29.00,34.00) years], and the differences were statistically significant ( P=0.022, P=0.020, P=0.009). Univariate and multivariate logistic regression analyses showed that days of blastocyst development, degree of blastocyst expansion, inner cell mass and trophectoderm grade did not increase the risk of chromosomal karyotype abnormalities of chorionic villi (all P>0.05). Conclusion:There is no significant correlation between blastocyst morphological evaluation parameters and chromosomal karyotype abnormalities in chorionic villi of missed abortion after fresh/frozen-thawed single blastocyst transfer with IVF/ICSI treatment.

Objective:Analyzing the influence of morphological evaluation parameters of blastocysts, including days of blastocyst development [day 5 (D5) and day 6 (D6)], degree of blastocyst expansion (4, 5, 6), inner cell mass and trophectoderm grade, on the occurrence of chromosomal karyotype abnormalities of chorionic villi in missed abortion after in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) treatment and fresh/frozen-thawed single blastocyst transfer. Methods:The clinical data of patients with missed abortion after IVF/ICSI treatment and fresh/frozen-thawed single blastocyst transfer from February 2015 to February 2023 in the Reproductive Center of the Third Affiliated Hospital of Zhengzhou University were included. Using a case-control study, the data were divided into two groups according to the detection results of chromosomal copy number variations (CNVs) in chorionic villi of missed abortion abnormal karyotype group ( n=139) and normal karyotype group ( n=82). The baseline data between the two groups were compared. Univariate logistic regression was used to investigate the effect of blastocyst morphological rating parameters on the occurrence of chromosomal karyotype abnormalities of chorionic villi in aborted tissues, and multivariate logistic regression was also used to adjust confounding factors. Results:Male age [(34.12±6.49) years], sperm morphology rate [5.00 (4.00,6.00)%] and female age [33.00 (30.00, 37.00) years] in abnormal karyotype group were higher than those in the normal karyotype group [(32.38±4.69) years, 4.00 (2.00,5.00)% and 31.50 (29.00,34.00) years], and the differences were statistically significant ( P=0.022, P=0.020, P=0.009). Univariate and multivariate logistic regression analyses showed that days of blastocyst development, degree of blastocyst expansion, inner cell mass and trophectoderm grade did not increase the risk of chromosomal karyotype abnormalities of chorionic villi (all P>0.05). Conclusion:There is no significant correlation between blastocyst morphological evaluation parameters and chromosomal karyotype abnormalities in chorionic villi of missed abortion after fresh/frozen-thawed single blastocyst transfer with IVF/ICSI treatment.

Objective: To explore the activity of auranofin against ovarian cancer cells and its possible molecular mechanism． Methods : The dose-response survival curve and IC50 of auranofin on ovarian cancer cell lines ，SKOV3，Caov3 and SW626 cells and immortalized normal human embryonic kidney HEK-293T cells were determined by CCK-8 method．Cell cycle was determined by flow cytometry．The levels of total glutathione ( GSH) ，reduced GSH and glutathione disulfide ( GSSG) ，thioredoxin reductase (TrxR) and reactive oxygen species (ROS) in cells were determined by microplate reader，and the reduced GSH / GSSG ratio was calculated．Western blot was used to determine the expression of cyclin dependent kinases( CDK) 4，CDK6，Cyclin D1，P53，p-P53 and MDM2 in SKOV3 and Caov3 ovarian cancer cells. Results : Compared with HEK-293T cells，the dose-response survival curves and IC50 values of SKOV3，Caov3 and SW626 cells showed that ovarian cancer cells were more sensitive to auranofin (P＜0. 05) ．After SKOV3 and Caov3 cells were treated with the dose of respective IC50 concentrations of auranofin，compared with the untreated cells group，the Auranofin IC50 group cells' intracellular levels of GSH，the ratio of reduced GSH / GSSG and the activity of TrxR decreased (t = 25. 11 /31. 18，14. 72 /19. 92，43. 30 /10. 74， all P＜0. 05) ，and the levels of ROS increased (t = 23. 82 /27. 71，P＜0. 05) ; cells number at G0 / G1 phases increased，with cells number at S and G2 phases decreased (P＜0. 05) ; and the expression levels of cell cycle-related proteins CDK4，CDK6，Cyclin D1 and the P53-specific E3 ubiquitin ligase MDM2 were down-regulated (t = 7. 51 /15. 59，17. 32 /11. 26，20. 78 /20. 78，24. 25 /17. 32，all P＜0. 05) ，while the expression levels of P53 and p-P53 were up-regulated (t = 17. 32 /24. 25，12. 12 /10. 39，all P ＜0. 05) ． Conclusion Auranofin causes oxidative stress in ovarian cancer cells by inhibiting TrxR activity，and by partially degrading MDM2 to stabilize and acti- vate P53，so as to block the cancer cells in G0 / G1 phase，and exert anti-ovarian cancer activities.