Nantong Maternity and Child Health Hospital

The main functions of testes are sperm production and androgen secretion in testicular befitting microenvironment. Excessive level of the reactive oxygen species (ROS) from metabolism and cellular events can lead to oxidative stress (OS) and ferroptosis, which injure the functions of mitochondria and endoplasmic reticulum (ER) of testicular cells, leading to the impaired spermatogenesis and spermiogenesis, as well as insufficient androgen. Our previous studies have shown that peroxiredoxin 4 (Prdx4) is a vital ER-located protector against endoplasmic reticulum stress (ERS) in ovarian granulosa cells. In this study, we explored whether Prdx4 played a beneficial role in testes, and the potential application value in male reproduction. Prdx4 knockout mice and Prdx4-/y Leydig cells were established using CRISPR/Cas9 system, the natural adult (9 weeks) mice, aging (9 months) mice, and wild-type MLTC-1 cells were used as control. To further investigate the protective effect of Prdx4 in the adult mice, the testicular 43°C water-bath was used to induce an OS model. The male Prdx4-/y mice aged over 9 months showed the age-related pathology of seminiferous tubule, the increased OS and ERS, and the decreased fertility. The adult Prdx4-/y mice under heat stress manifested increased levels of OS, including almost 1.5-fold rise in 4-hydroxynonenal (4-HNE) and 8-hydroxy-2'- deoxyguanosine (8-OHdG), higher ferroptosis, and poorer fertility. Prdx4-/y Leydig cells showed a higher level of ferroptosis after hydrogen peroxide treatment. ACSL4 increased by 18.6% while COX2 increased by 13.5%, and the expressions of GPX4 and SLC7A11 decreased by 49.2% and 24.3%, respectively. Increased levels of ferroptosis in MLTC-1 leads to mitochondria-derived OS and apoptosis, which can be partially restored by deferoxamine and overexpressing SLC7A11. We concluded that Prdx4 as a protective factor plays a critical role in preserving spermatogenesis by alleviating testicular OS and ferroptosis, and that Prdx4 may be known as a potential target in mitigating the adverse effects of natural aging and OS injury.

With the nationwide relaxation of controls for the COVID-19 epidemic from December 2022 in China, fertility and in vitro fertilization (IVF) centers reported increasing numbers of infected patients in a short time. However, there is a lack of information regarding the effects of SARS-CoV-2 infection on pregnancy outcomes after IVF–embryo transfer (ET)/intracytoplasmic sperm injection (ICSI)–ET/frozen–thawed embryo transfer (FET), especially the live birth rate. This multicenter retrospective study was conducted at three public IVF centers in Jiangsu province of China. Women aged 20–42 years who were undergoing IVF–ET/ICSI–ET/FET treatment were consecutively registered from November 23, 2022 to January 7, 2023, with follow-up until October, 2023. Patients during the same period in the previous year were included in the control group. Each woman contributed only one transplantation cycle. Multivariate regression analysis was used to evaluate clinical outcomes primarily between different policy period other than individual infected or not. Covariates included demographic characteristics (age, body mass index (BMI), infertility cause, anti-müllerian hormone (AMH), and Antral Follicle Counting(AFC)) as well as cycle characteristics (treatment protocol, endometrial thickness, number of embryos transferred, and embryonic morphology). In this study, we included 779 women in the FET group and 168 women in the IVF–ET/ICSI–ET group, analyze the clinical outcomes of each group separately before and after the relaxation of controls for the COVID-19 epidemic. Pregnancy outcomes, including the clinical pregnancy rate, miscarriage rate, ongoing pregnancy rate, multiple pregnancy rate, and live birth rate, did not change significantly in different policy period. However, the rate of gestational diabetes mellitus (GDM) in FET cycles rather than IVF–ET/ICSI–ET cycles after the relaxation of controls was higher than that 1 year earlier (17.8% vs. 6.8%). The findings of this study suggest that the relaxation of controls for the COVID-19 epidemic does not have a clear negative effect on the pregnancy outcomes after ET, despite current clinical concerns. Nevertheless, the morbidity of gestational diabetes in FET cycles after the relaxation of controls warrants greater attention.

Enterococcus faecalis promotes colorectal cancer (CRC) in murine models through the microbiota-induced bystander effect (MIBE), but its potential to induce epigenetic changes remains unexplored. This study investigates how E. faecalis-infected macrophages induce epigenetic alterations contributing to CRC pathogenesis. RAW264.7 macrophages were infected with E. faecalis OG1RF (superoxide-producing), WY84 (superoxide-deficient), or PBS. Conditioned media were collected and used to treat IEC-6 intestinal epithelial cells. Genome-wide H3 acetylation was assessed using chromatin immunoprecipitation sequencing (ChIP-seq). Western blotting and immunofluorescence staining were employed to investigate E. faecalis-induced histone H3 acetylation and associated signaling pathways. Furthermore, colonic organoid cultures and E. faecalis-colonized Il10−/− mouse models were utilized to validate the in vitro findings. Conditioned medium (CM) from E. faecalis-infected macrophages significantly increased histone H3 acetylation at lysines 27 and 9 (H3K27ac/H3K9ac) in IEC-6 intestinal epithelial cells and murine colonic organoids, with similar findings validated in vivo in E. faecalis-colonized Il10−/− mice. ChIP-seq revealed genome-wide alterations in H3K27ac in cells treated with CM from E. faecalis-infected macrophages, leading to activation of CRC-associated signaling pathways. Metabolomic analysis identified 12-hydroxyeicosatetraenoic acid (12-HETE), produced by E. faecalis-infected macrophages, as a mediator of bystander effect promoting H3 acetylation. Furthermore, CM from E. faecalis-infected macrophages and 12-HETE decreased E-cadherin and increased vimentin expression in IEC-6 cells, indicating epithelial-mesenchymal transition—a finding corroborated by reduced E-cadherin levels in intestinal biopsies from E. faecalis-colonized Il10−/− mice. Our results indicate that E. faecalis-infected macrophages produce 12-HETE, which promotes epigenetic alterations and induces EMT in intestinal epithelial cells via a bystander effect. These findings strengthen the link between gut microbiota and innate immunity in CRC development, highlighting potential targets for innovative prevention and therapeutic strategies. Not applicable.

Endoscopic resection (ER) has been successfully used to remove giant esophageal submucosal tumors (SMTs). However, though rare, esophageal fistula can also develop after endoscopic resection. The aim of this study was to evaluate risk factors for fistula formation after ER of giant esophageal SMTs. This retrospective study enrolled patients with huge esophageal SMTs who underwent ER. We collected patient demographics, lesion-related factors, and procedural outcomes data and explored the correlation between esophageal fistula formation and damage to perforating vessels. Of 174 patients, 17 (9.8%) had esophageal fistula. The multivariate analysis demonstrated that perforating vessel damage and full-thickness muscular injury were significant risk factors for esophageal fistula. Then, we used a scoring system to predict esophageal fistula and categorized patients into low-risk (0-2) and high-risk (3-6) groups. Patients with high scores were more likely to suffer esophageal fistula. Our score model presented excellent discriminatory power in the internal validation (AUC: 0.932; 95% CI: 0.797-1.000) and goodness-of-fit in the Hosmer-Lemeshow test (p=0.748). Fistula formation after ER of giant esophageal SMTs is associated with damage to perforating vessels.

Intrahepatic cholestasis of pregnancy (ICP) is a common pregnancy-specific disorder characterized by pruritus and elevated serum total bile acids (TBA), and is associated with adverse outcomes such as preterm birth. This study aimed to investigate the role of autotaxin (ATX) in ICP-related pruritus and preterm labor. This prospective cohort study included 180 pregnant women (100 diagnosed with ICP and 80 healthy controls) recruited from the Affiliated Maternal and Child Health Care Hospital of Nantong University between 2024 and 2025. Serum TBA, ATX, pruritus scores, and other relevant biochemical markers were measured and compared between patients with ICP and controls. Perinatal outcomes, including gestational age at delivery, birth weight, and Apgar scores at 1 and 5 minutes, were recorded for all participants. We evaluated the association of ATX and TBA with pruritus scores and preterm delivery. Statistical analyses were performed using t-tests, regression models, and Mann–Whitney U tests, as appropriate. No demographic differences were observed between the two groups (p > 0.05). Patients with ICP exhibited significantly higher TBA, Alanine Aminotransferase (ALT), Aspartate Aminotransferase (AST), ATX concentrations, and pruritus scores (p < 0.05). The median ATX level was 8.08 ng/mL in the ICP group versus 3.45 ng/mL in controls. Preterm delivery and lower birth weight were more prevalent among ICP patients (p < 0.05), while Apgar scores were similar (p > 0.05). Maternal serum ATX concentrations were positively associated with pruritus in ICP patients. Although TBA, ALT, and AST concentrations were higher in preterm ICP cases (p < 0.05), ATX concentrations did not differ between preterm and term deliveries in ICP patients (p > 0.05) and had no significant correlation with preterm birth (p > 0.05). In summary, serum ATX concentrations were increased in early-onset ICP, correlating positively with pruritus severity but not with preterm birth.

BackgroundBreast cancer (BC) is a major health concern among women. SanHuang decoction (SHD), a Chinese herbal formula, has exhibited the potential for clinical application in patients with BC. This study aimed to clarify the mechanism of SHD’s action in the treatment of BC based on macrophage function and nuclear factor kappa B (NF-κB) pathway.MethodsM1 polarized macrophages were cocultured with MDA-MB-231 BC cells, and treated with 10 mg/mL of SHD for 72 hours. Scratch and transwell assays were used to observe the inhibitory effect of SHD, and the levels of interleukin-6 (IL-6), tumor necrosis factor alpha (TNF-α), IL-1β, cleaved caspase 3, B-cell lymphoma 2 (BCL-2), BCL-2-associated X (BAX), P65, phosphorylated P65 (p-P65), inhibitor of kappa B (IκB), and phosphorylated IκB (p-IκB) were detected via Western blotting. Forty patients with BC were randomly divided into a control group [neoadjuvant chemotherapy (NACT)] and observation group (NACT + SHD). After 9 weeks of treatment, magnetic resonance imaging scans were performed to evaluate the tumor size, and immunofluorescence staining was performed to investigate the effect of SHD on the regulation of M1 macrophages in tumor tissue.ResultsSHD inhibited the migration ability of MDA-MB-231 cells, and the effect was significantly enhanced after coculture with M1/THP-1. Western blot results showed that SHD could significantly increase the expression of TNF-α, IL-1β, IL-6, BAX, cleaved caspase 3, p-P65, and p-IκB while decreasing that of BCL-2. In vivo studies showed that SHD could reduce the tumor size and increase the expression of M1 macrophages.ConclusionsSHD could enhance the expression of M1/THP-1 cytokines, promote the inflammatory response in the tumor microenvironment, and thus inhibit the proliferation of BC and regulate its pathological process, likely through activating NF-κB signaling pathway.