To investigate whether long-term low-dose exposure to polystyrene microplastics (PS-MPs) induces ferroptosis in testicular Sertoli cells and then leads to testicular injury.

Forty 8-week-old male C57BL/6 mice were randomly divided into a control group (deionized water group) and a PS-MPs group [2. 5 mg/(kg·d)], with 20 mice in each group. Corresponding agents were gavaged once a day for 12 consecutive months. HE staining and Prussian blue staining were used to detect histopathological damage and accumulation of ferrous ions in the testes. Electron transmission microscopy was employed to observe the mitochondrial morphology of testicular Sertoli cells. Mouse Sertoli cell line TM4 was divided into a Con group (standard culture) and an MPs group (2. 5 μg/mL PS-MPs). After both groups underwent continuous exposure and passed up to the 100th generation, morphological changes were observed under the microscope; cell viability was detected with CCK8 assay, and production of reactive oxygen species (ROS) and mitochondrial membrane potential (MMP) were detected with a ROS probe and a mitochondrial membrane potential probe (JC-1), respectively. Flow cytometry, ferrous ion (Fe²⁺) kit and Western blotting were applied to detect cell apoptosis, intracellular iron ion content, and protein levels of key molecules of ferroptosis in tissues and cells.

Long-term exposure to PS-MPs resulted in significantly reduced diameter and thickness of mouse varicocele (P<0. 01), fewer testicular Sertoli cells (P<0. 05), with characteristic ferroptosis alterations in the mitochondria, and increased accumulation of ferrous ions in testicular tissue. Exposure to PS-MPs down-regulated the key molecules of ferroptosis, glutathione peroxidase 4(GPX4) and ferritin light chain (FTL) when compared with the control group (P<0. 05). In the cell model, long-term PS-MPs exposure led to morphological changes and decreased cell viability (P<0. 05), more production of ROS (P<0. 01), and decrease in MMP (P<0. 05) of TM4 cells. The exposure had no effect on cell apoptosis, but elevated the intracellular content of ferric ions (P<0. 01), and down-regulated GPX4 and FTL protein levels (P<0. 05).

Long-term low-dose exposure to PS-MPs induces mitochondrial damage and oxidative stress in testicular Sertoli cells, activates the ferroptosis pathway, and ultimately leads to testicular injury in mice.

To investigate the effects and potential mechanisms of polystyrene micro/nanoplastics (PS-MNPs) on testicular Sertoli cells.

Sixty male C57BL/6N mice (8 weeks old) were randomly divided into a control group (deionized water), a PS-NPs group [particle size of 20 nm, 2. 5 mg/(kg·d)], and a PS-MPs group [particle size of 5 μm, 2. 5 mg/(kg·d)], with 20 mice in each group. The corresponding agents were gavaged once daily for 6 months. HE staining was used to observe the histopathological and morphological changes in the testicular tissues. Immunohistochemistry of marker proteins was employed to evaluate the changes in the number of Sertoli cells. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed to identify functions and signaling pathways enriched in the testicular transcriptome. Mouse testicular Sertoli cell line TM4 was divided into a control group (deionized water), a 2. 5NPs group (2. 5 μg/mL), and a 2. 5MPs group (2. 5 μg/mL). All groups received continuous exposure through 130 cell passages. Cell viability and proliferative capacity were evaluated using CCK-8 assay and EdU incorporation, while cell migration was assessed using transwell and cell scratch assays. RT-qPCR and Western blotting were used to detect the changes in the expression of key molecules regulating mesenchymal phenotypic transformation (MPT) at mRNA and protein levels.

Pathological analysis revealed that, when compared to the control group, PS-NPs and PS-MPs treatment resulted in extended spaces between testicular seminiferous tubules, loosely arranged spermatogenic cells, and enhanced vacuolization. Immunohistochemical analysis of marker proteins indicated a decreasing trend in the number of testicular Sertoli cells in the PS-NPs and PS-MPs groups than the control group, with the PS-NPs group having statistical significance (P<0. 01). GO and KEGG enrichment analyses revealed that PS-MNPs exposure-related altered genes were significantly enriched in cell adhesion signaling pathways (P<0. 05). PS-MPs exposure significantly inhibited the growth and migration ability of TM4 cells (P<0. 05), but PS-NPs exposure had no such effect on cell growth but notably enhanced cell migration ability. PS-NPs exposure inhibited the expression levels of E-cadherin and ZO-1 (P<0. 01) and up-regulated the expression of N-cadherin and vimentin (P<0. 01), and PS-MPs exposure led to significant up-regulation of vimentin (P<0. 01) and down-regulation of E-cadherin, N-cadherin, and ZO-1 (P<0. 05). Both PS-MPs and PS-NPs exposure up-regulated the mRNA levels of Snail2, Twist1, and Zeb2 (P<0. 01).

Exposure of PS-MNPs leads to abnormal proliferation and migration of TM4 cells, induces decreases in cell-cell contacts among Sertoli cells and spermatogenic cells at all levels possibly through MPT, and thus results in testicular damage.

Non‐small cell lung cancer (NSCLC), including the lung squamous cell carcinoma (LUSC) and lung adenocarcinoma (LUAD) subtypes, is a malignant tumor type with a poor 5‐year survival rate. The identification of new powerful diagnostic biomarkers, prognostic biomarkers, and potential therapeutic targets in NSCLC is urgently required.

The UCSC Xena, UALCAN, and GEO databases were used to screen and analyze differentially expressed genes, regulatory modes, and genetic/epigenetic alterations in NSCLC. The UCSC Xena database, GEO database, tissue microarray, and immunohistochemistry staining analyses were used to evaluate the diagnostic and prognostic values. Gain‐of‐function assays were performed to examine the roles. The ESTIMATE, TIMER, Linked Omics, STRING, and DAVID algorithms were used to analyze potential molecular mechanisms.

NR3C2 was identified as a potentially important molecule in NSCLC. NR3C2 is expressed at low levels in NSCLC, LUAD, and LUSC tissues, which is significantly related to the clinical indexes of these patients. Receiver operating characteristic curve analysis suggests that the altered NR3C2 expression patterns have diagnostic value in NSCLC, LUAD, and especially LUSC patients. Decreased NR3C2 expression levels can help predict poor prognosis in NSCLC and LUAD patients but not in LUSC patients. These results have been confirmed both with database analysis and real‐world clinical samples on a tissue microarray. Copy number variation contributes to low NR3C2 expression levels in NSCLC and LUAD, while promoter DNA methylation is involved in its downregulation in LUSC. Two NR3C2 promoter methylation sites have high sensitivity and specificity for LUSC diagnosis with clinical application potential. NR3C2 may be a key participant in NSCLC development and progression and is closely associated with the tumor microenvironment and immune cell infiltration. NR3C2 co‐expressed genes are involved in many cancer‐related signaling pathways, further supporting a potentially significant role of NR3C2 in NSCLC.

NR3C2 is a novel potential diagnostic and prognostic biomarker and therapeutic target in NSCLC.

Breast cancer (BRCA) is the most common malignant tumor among women worldwide. Despite advances in treatment, many patients still die from a lack of effective diagnostic and prognostic markers and powerful therapeutic targets. LIM homeobox genes (LHXs) play vital roles in regulating the development of various organisms. However, there are limited reports regarding their roles in the diagnosis, prognosis, and treatment of BRCA.

UALCAN, Kaplan–Meier plotter, cBioPortal, GeneMANIA, STRING, DAVID 6.8, TRRUST v2, LinkedOmics, and TIMER were utilized to analyze differential expression, prognostic value, genetic alteration, neighbor gene network, transcription factor targets, kinase targets, and immune cell infiltration of LHXs in BRCA patients.

LHX gene expression patterns are clear in BRCA and its different subtypes. Further analyses indicated that this altered expression is possibly affected by genetic and/or epigenetic changes. The prognostic and diagnostic values of certain LHXs are unique to different BRCA subtypes. LHXs are mainly involved in the regulation of differentiation and development, and their neighbor genes are primarily involved in cancer‐related pathways. Moreover, most LHXs are closely correlated with immune cell infiltration. Furthermore, LHXs may exert their functions by regulating a series of transcription factor and kinase targets.

LHXs are unique diagnostic and prognostic markers and participate in cancer through different signaling pathways and/or regulatory mechanisms in BRCA. This study provides potential applications of LHXs for the diagnosis, prognosis, and treatment of BRCA and its different subtypes.