Ovarian aging, characterized by declining ovarian reserve, is a pacemaker of aging in the female body. Oxidative stress leads to apoptosis, mitochondrial dysfunction, inflammation, and telomere shortening, accelerating ovarian aging. Scavenging reactive oxygen species (ROS) has been shown to delay ovarian aging; however, there remains a significant lack of antioxidants with both proven efficacy and minimal side effects. DNA tetrahedral nanostructure (DTN) is a promising nucleic acid framework with antioxidant and anti-apoptotic properties. We developed FSH-DTN, a modified nanoparticle equipped with a follicle-stimulating hormone receptor-targeting peptide (FSH33-53) to enhance ovarian accumulation. Compared to native DTN, FSH-DTN showed superior ovarian targeting efficiency as confirmed by in vivo imaging. In both in vivo and in vitro models of acute, subacute, and chronic ovarian aging, FSH-DTN demonstrated superior antioxidant, anti-apoptotic, and anti-aging effects. Further investigation revealed that FSH-DTN can directly eliminate ROS in the ovaries while enhancing ovarian antioxidant capacity by activating the NRF2 signaling pathway, thereby protecting ovarian function. In this study, we offer a new strategy for neutralizing oxidative stress to delay ovarian aging.

Recent studies have highlighted the crucial role of mechanical properties in the ovarian microenvironment for ovarian function. However, the mechanisms that cause ovarian matrix stiffening during aging remain incompletely understood. Here we utilized atomic force microscopy (AFM) to demonstrate that human ovarian matrix stiffness increases with aging and in pathophysiological conditions, such as chemotherapy-induced premature ovarian insufficiency (POI), polycystic ovary syndrome (PCOS) and ovarian endometriosis. By integrating proteomic analysis of human ovarian tissue with transcriptomic profiling of human ovarian fibroblasts, we identified that IL-11, which is elevated in aging ovaries of mice, rats and humans, activates fibroblasts to secrete extracellular matrix (ECM), thereby increasing ovarian matrix stiffness. Genetic deletion of Il11ra1 in mice mitigated the increase in ovarian matrix stiffness and the decline in ovarian function associated with aging, chemotherapy-induced POI and PCOS. Single-nuclei RNA sequencing (snRNA-seq) revealed that blocking Il11ra1 reduces the proportion of activated fibroblasts. Furthermore, administration of siIl11 nanoparticles to aged mice and rats enhanced fertility and reduced ovarian matrix stiffness. Together, these findings highlight the pro-inflammatory factor IL-11 in regulating ovarian matrix stiffness. We propose that anti-IL-11 therapy represents a promising translational strategy for delaying ovarian aging.

INTRODUCTION:Carbon-based nanomaterials have attracted increasing attention due to their routine exposure and potential health risks. However, the impact of carbon-based nanomaterials on ovarian function remains poorly understood. OBJECTIVE:This study aimed to systematically evaluate the ovarian toxicity of three representative carbon-based nanomaterials and to elucidate the underlying mechanisms of ovarian dysfunction. METHODS:Female mice were exposed to multi-walled carbon nanotubes (MWCNTs), graphene, or fullerene. Estrous cycle, hormone levels, and follicular development were assessed. Transcriptomic sequencing and molecular analyses were performed to explore potential mechanisms, and the protective effect of NLRP3 inhibition was validated using mouse, granulosa cell, and human ovarian cortex models. RESULTS:All three carbon-based nanomaterials induced estrous cycle disruption in mice. Hormone analysis revealed elevated serum follicle-stimulating hormone levels in all exposure groups, while a significant decrease in estradiol was observed only in MWCNTs group. MWCNTs and graphene significantly reduced the numbers of primordial and growing follicles and increased the number of atretic follicles. In contrast, the fullerene exhibited milder effects, with only an increase in atretic follicles. None of the three nanoparticles significantly affected female fertility in mice. Further transcriptomic analysis identified the NLRP3 inflammasome as a key mediator of granulosa cell injury. Unlike classical apoptotic pathways that mediate granulosa cell death, exposure to carbon-based nanomaterials induced pyroptosis in granulosa cells by activating the NLRP3 inflammasome pathway, subsequently leading to follicular atresia. Notably, intervention experiments using granulosa cells, mouse model, and cultured human ovarian cortex demonstrated that the NLRP3 inhibitor MCC950 effectively alleviated carbon-based nanomaterials induced ovarian dysfunction. CONCLUSION:These findings reveal a novel mechanism by which carbon-based nanomaterials impair ovarian function through NLRP3 inflammasome-mediated pyroptosis and highlight NLRP3 inhibition as a potential therapeutic strategy for mitigating carbon-based nanomaterials associated ovarian injury.

Deterministic lateral displacement (DLD) is a size-based microfluidic technology that allows for the continuous separation of suspended particles. Nevertheless, the shape and deformability affect the critical diameter, especially in terms of bio-samples. In this study, the influence of the shape and deformability of red blood cells (RBCs) on the critical diameter was investigated in DLD devices, leading to the design of microfluidic chips with multiple critical diameters to separate the blood samples. A biconcave disk-shaped RBC model was constructed in the simulation of the sorting process, and the Neo-Hookean model was applied to describe the deformability. According to the simulation results, polystyrene particles, spherocytes, and normal RBCs were employed, and the effectiveness of the design was confirmed by the experiments. Results indicated that increased deformability corresponded to an increase in critical diameter. Because of the increased rigidity of the membrane structure, spherocytes flowed out of the desired outlet in a typical zig-zag pattern with a sorting purity of 92.97%, while the removal rate of the healthy RBCs was 95.58%, which achieves the separation of healthy RBCs and spherocytes in the whole blood samples.

Ovarian aging is considered to be the pacemaker of female aging, and is linked to various comorbidities such as osteoporosis, cardiovascular diseases, and cognitive decline. Many efforts have been made to determine the mechanisms underlying ovarian aging, but their potential to act as hallmarks to predict and intervene in this process currently remains unclear. In this review we propose nine hallmarks as common features of ovarian aging: genomic instability, telomere attrition, epigenetic alterations, impaired autophagy, cellular senescence, deregulated nutrient-sensing, mitochondrial dysfunction, oxidative stress, and chronic inflammation. Understanding the interaction between these hallmarks poses a significant challenge but may also pave the way to the identification of pharmaceutical targets that can attenuate ovarian aging.

Abnormal apoptosis both maintains endometrial cell growth and induces endometrial pathogenesis. The etiology of endometriosis is unclear and no treatment is curative. Therefore, the aim herein was to identify genes involved in the pathogenesis of endometriosis. Using the data from our previous results and RNA sequencing data of normal endometrial tissue and ovarian endometrioma (OMA) tissue, along with Gene Expression Omnibus (GEO) dataset on endometriosis, we identified an apoptotic-related gene, meis homeobox I (MEIS1). Normal endometrium, eutopic endometrium and ectopic endometriotic tissues were used to detect MEIS1. Primary normal endometrial and eutopic endometrial stromal cells were isolated and cultured for exploring the function of MEIS1 and related pathways. A mouse endometriosis model was used to verify the therapeutic effects of MEIS1. The mRNA and protein of MEIS1 in tissues from patients with endometriosis were decreased. Overexpression of MEIS1 induced the apoptosis of primary eutopic endometrium stromal cells by regulating TNFR1. Using Cell Counting Kit 8 (CCK8) assay and EdU assay, we found that knockdown of MEIS1 promoted the proliferation of primary normal endometrium stromal cells. We also observe that upregulated MEIS1 may lead to caspase pathway activation, promoting endometrial cell apoptosis. Furthermore, MEIS1 lentivirus inhibited endometriotic lesion formation and induced apoptosis in the mouse endometriosis model. These cumulative findings suggest that MEIS1 may mediate apoptosis by initiating TNFR1 in endometrial cells via the caspase pathway.

Exosomes are a type of extracellular vesicle ranging from 30 to 150 nm in diameter that contain constituents of the cells that secrete them. The initial challenge in utilizing exosomes lies in their effective sorting from body fluids. However, traditional methods for exosome sorting are often time-consuming and costly. In this work, a cascaded microfluidic system was proposed for sorting exosomes from whole blood samples by integrating a deterministic lateral displacement (DLD) module with a circulation tangential flow filtration (CTFF) module. The DLD module, featuring multiple critical diameters, served as the first stage to remove blood cells from the sample. The CTFF module functioned as the second stage for exosome sorting. Sorting parameters were optimized including the sample flux, dilution factor of the blood sample, and transmembrane pressure. Exosomes were efficiently sorted from the plasma samples using the CTFF chip, achieving a recovery rate of 89.64% and a recovery purity of 84.27%. When employed to directly sort exosomes from whole blood samples, this cascaded system achieved a recovery purity of 70.21% and a recovery rate of 28.64%. The proposed sorting system presents a promising approach for the efficient sorting of micro/nanoparticles in complex biological samples.

Liver fibrosis, a hallmark pathological endpoint of chronic aging-related liver diseases, remains a clinical challenge with limited therapeutic options. In healthy liver, myeloid cells constitute <5% of total hepatic immune cells, primarily comprising tissue-resident Kupffer cells. However, during aging or chronic injury, bone marrow-derived myeloid cell recruitment increases by two- to threefold in murine fibrotic models, reaching 15%-20% of intrahepatic immune populations. These infiltrating myeloid subsets exhibit functional plasticity, dynamically differentiating into pro-inflammatory macrophages or fibrosis-promoting Kupffer-like cells, contingent upon chemokine gradients (e.g., CCL2/CCR2 axis) and damage-associated molecular patterns (DAMPs). This review systematically examines the regulatory mechanisms of myeloid cells in liver fibrogenesis, with particular emphasis on their developmental origins, hepatic recruitment dynamics, functional heterogeneity, and pathogenic contributions to fibrosis. Furthermore, signaling pathways involving myeloid cells in liver fibrosis and therapeutic approaches modulating their differentiation and recruitment are discussed in this review.

With increasing proportion of the elderly in the population, age-related diseases (ARD) lead to a considerable healthcare burden to society. Prevention and treatment of ARD can decrease the negative impact of aging and the burden of disease. The aging rate is closely associated with the production of high levels of reactive oxygen species (ROS). ROS-mediated oxidative stress in aging triggers aging-related changes through lipid peroxidation, protein oxidation, and DNA oxidation. Antioxidants can control autoxidation by scavenging free radicals or inhibiting their formation, thereby reducing oxidative stress. Benefiting from significant advances in nanotechnology, a large number of nanomaterials with ROS-scavenging capabilities have been developed. ROS-scavenging nanomaterials can be divided into two categories: nanomaterials as carriers for delivering ROS-scavenging drugs, and nanomaterials themselves with ROS-scavenging activity. This study summarizes the current advances in ROS-scavenging nanomaterials for prevention and treatment of ARD, highlights the potential mechanisms of the nanomaterials used and discusses the challenges and prospects for their applications.

BACKGROUND Chemotherapy-associated ovarian damage (CAOD) is one of the most feared short- and long-term side effects of anticancer treatment in premenopausal women. Accumulating detailed data show that different chemotherapy regimens can lead to disturbance of ovarian hormone levels, reduced or lost fertility, and an increased risk of early menopause. Previous studies have often focused on the direct effects of chemotherapeutic drugs on ovarian follicles, such as direct DNA damage-mediated apoptotic death and primordial follicle burnout. Emerging evidence has revealed an imbalance in the ovarian microenvironment during chemotherapy. The ovarian microenvironment provides nutritional support and transportation of signals that stimulate the growth and development of follicles, ovulation, and corpus luteum formation. The close interaction between the ovarian microenvironment and follicles can determine ovarian function. Therefore, designing novel and precise strategies to manipulate the ovarian microenvironment may be a new strategy to protect ovarian function during chemotherapy.OBJECTIVE AND RATIONALE This review details the changes that occur in the ovarian microenvironment during chemotherapy and emphasizes the importance of developing new therapeutics that protect ovarian function by targeting the ovarian microenvironment during chemotherapy.SEARCH METHODS A comprehensive review of the literature was performed by searching PubMed up to April 2024. Search terms included 'ovarian microenvironment' (ovarian extracellular matrix, ovarian stromal cells, ovarian interstitial, ovarian blood vessels, ovarian lymphatic vessels, ovarian macrophages, ovarian lymphocytes, ovarian immune cytokines, ovarian oxidative stress, ovarian reactive oxygen species, ovarian senescence cells, ovarian senescence-associated secretory phenotypes, ovarian oogonial stem cells, ovarian stem cells), terms related to ovarian function (reproductive health, fertility, infertility, fecundity, ovarian reserve, ovarian function, menopause, decreased ovarian reserve, premature ovarian insufficiency/failure), and terms related to chemotherapy (cyclophosphamide, lfosfamide, chlormethine, chlorambucil, busulfan, melphalan, procarbazine, cisplatin, doxorubicin, carboplatin, taxane, paclitaxel, docetaxel, 5-fluorouraci, vincristine, methotrexate, dactinomycin, bleomycin, mercaptopurine).OUTCOMES The ovarian microenvironment shows great changes during chemotherapy, inducing extracellular matrix deposition and stromal fibrosis, angiogenesis disorders, immune microenvironment disturbance, oxidative stress imbalances, ovarian stem cell exhaustion, and cell senescence, thereby lowering the quantity and quality of ovarian follicles. Several methods targeting the ovarian microenvironment have been adopted to prevent and treat CAOD, such as stem cell therapy and the use of free radical scavengers, senolytherapies, immunomodulators, and proangiogenic factors.WIDER IMPLICATIONS Ovarian function is determined by its 'seeds' (follicles) and 'soil' (ovarian microenvironment). The ovarian microenvironment has been reported to play a vital role in CAOD and targeting the ovarian microenvironment may present potential therapeutic approaches for CAOD. However, the relation between the ovarian microenvironment, its regulatory networks, and CAOD needs to be further studied. A better understanding of these issues could be helpful in explaining the pathogenesis of CAOD and creating innovative strategies for counteracting the effects exerted on ovarian function. Our aim is that this narrative review of CAOD will stimulate more research in this important field.REGISTRATION NUMBER Not applicable. Graphical Abstract Chemotherapy causes an imbalance of the ovarian microenvironment leading to chemotherapy-associated ovarian damage and dysfunction, and further research is needed to explore the possible protective treatments. Created with BioRender.com, with permission.

Limited understanding exists regarding how aging impacts the cellular and molecular aspects of the human ovary. This study combines single-cell RNA sequencing and spatial transcriptomics to systematically characterize human ovarian aging. Spatiotemporal molecular signatures of the eight types of ovarian cells during aging are observed. An analysis of age-associated changes in gene expression reveals that DNA damage response may be a key biological pathway in oocyte aging. Three granulosa cells subtypes and five theca and stromal cells subtypes, as well as their spatiotemporal transcriptomics changes during aging, are identified. FOXP1 emerges as a regulator of ovarian aging, declining with age and inhibiting CDKN1A transcription. Silencing FOXP1 results in premature ovarian insufficiency in mice. These findings offer a comprehensive understanding of spatiotemporal variability in human ovarian aging, aiding the prioritization of potential diagnostic biomarkers and therapeutic strategies. Ovarian aging has an important role in health and fertility; however, the molecular mechanisms underlying it remain incompletely understood. Here the authors use single-cell and spatial transcriptomics in reproductively young, middle-aged and older human ovarian tissue to elucidate ovarian aging. They describe spatiotemporal changes in ovarian cells and highlight the important regulatory role of FOXP1.

Ovarian aging is marked by a reduction in the quantity and quality of ovarian follicles, leading to a decline in female fertility and ovarian endocrine function. While the biological characteristics of ovarian aging are well-established, the exact mechanisms underlying this process remain elusive. Recent studies underscore the vital role of trace elements (TEs) in maintaining ovarian function. Imbalances in TEs can lead to ovarian aging, characterized by reduced enzyme activity, hormonal imbalances, ovulatory disorders, and decreased fertility. A comprehensive understanding of the relationship between systemic and cellular TEs balance and ovarian aging is critical for developing treatments to delay aging and manage age-related conditions. This review consolidates current insights into TEs homeostasis and its impact on ovarian aging, assesses how altered TEs metabolism affects ovarian aging, and suggests future research directions to prolong ovarian reproductive life. These studies are expected to offer novel approaches for mitigating ovarian aging.

The human female reproductive lifespan significantly diminishes with age, leading to decreased fertility, reduced fertility quality and endocrine function disorders. While many aspects of aging in general have been extensively documented, the precise mechanisms governing programmed aging in the female reproductive system remain elusive. Recent advancements in omics technologies and computational capabilities have facilitated the emergence of multiomics deep phenotyping. Through the application and refinement of various high-throughput omics methods, a substantial volume of omics data has been generated, deepening our comprehension of the pathogenesis and molecular underpinnings of reproductive aging. This review highlights current and emerging multiomics approaches for investigating female reproductive aging, encompassing genomics, epigenomics, transcriptomics, proteomics, metabolomics, and microbiomics. We elucidate their influence on fundamental cell biology and translational research in the context of reproductive aging, address the limitations and current challenges associated with multiomics studies, and offer a glimpse into future prospects.

Abstract Background In systematic lymphadenectomy surgeries for gynecological malignancies, the potential disruption of ovarian lymphatic drainage function is a concern, particularly in individuals with a vested interest in fertility preservation. However, the current impact of lymphatic drainage disruption on ovarian function remains unclear and has not received sufficient attention from clinical practitioners. This study aims to elucidate the effects of ovarian lymphatic drainage obstruction on ovarian function in mice. Methods Evans blue dye was injected into the ovaries of mice to visualize ovarian draining lymphatic vessels (dLVs), which were subsequently ligated under the stereomicroscope, aiming to replicate the surgical injury to the ovarian lymphatic drainage during systematic lymphadenectomy. Serum estradiol, progesterone and anti-Müllerian hormone (AMH) levels were detected by ELISA, the number of follicles was counted by H&E staining, pregnancy rate and average litter sizes were evaluated through mating test, and the oocyte quality was assessed via spindle staining, ROS level detection, and Annexin-V staining. Additionally, immune cells and cytokines in the ovary were analyzed. Results Following intraovarian injection of Evans blue dye, the lymphatic vessels near the porta ovarii were blue, subsequently draining into the draining lymphatic nodes (dLNs) in the dorsal region of the upper pole of the kidney. At 30 days post dLV ligation, the ligation group exhibited a significant increase in ovarian weight and ovarian index compared to the control group. Obstruction of ovarian lymphatic drainage led to the decrease of serum progesterone and AMH levels by nearly 50%, the decrease of growing follicles and the increase of atretic follicles. The pregnancy rate decreased from 81.9–29.8% and the average litter size decreased from 5.6 to 3.3 per female in the ligation group. Furthermore, obstruction of ovarian lymphatic drainage elevated oxidative stress levels in oocytes, promoted oocyte apoptosis and meiosis arrest. These outcomes of impaired ovarian function may be attributed to the disturbance of the ovarian immune microenvironment, characterized by an increase of macrophages and the proinflammatory factors IL-6 in mouse ovaries. Conclusions The obstruction of ovarian lymphatic drainage leads to ovarian dysfunction, suggesting the importance of minimizing disruption to the ovarian lymphatic drainage pathway during lymphadenectomy, particularly in cases involving fertility preservation management.

Endosomes are characterized by the presence of various phosphoinositides that are essential for defining the membrane properties. However, the interplay between endosomal phosphoinositides metabolism and innate immunity is yet to be fully understood. Here, our findings highlight the evolutionary continuity of RAB-10/Rab10's involvement in regulating innate immunity. Upon infection of Caenorhabditis elegans with Pseudomonas aeruginosa, an increase in RAB-10 activity was observed in the intestine. Conversely, when RAB-10 was absent, the intestinal diacylglycerols (DAGs) decreased, and the animal's response to the pathogen was impaired. Further research revealed that UNC-16/JIP3 acts as an RAB-10 effector, facilitating the recruitment of phospholipase EGL-8 to endosomes. This leads to a decrease in endosomal phosphatidylinositol 4,5-bisphosphate (PI(4,5)P2) and an elevation of DAGs, as well as the activation of the PMK-1/p38 MAPK innate immune pathway. It is noteworthy that the dimerization of UNC-16 is a prerequisite for its interaction with RAB-10(GTP) and the recruitment of EGL-8. Moreover, we ascertained that the rise in RAB-10 activity, due to infection, was attributed to the augmented expression of LET-413/Erbin, and the nuclear receptor NHR-25/NR5A1/2 was determined to be indispensable for this increase. Hence, this study illuminates the significance of endosomal PI(4,5)P2 catabolism in boosting innate immunity and outlines an NHR-25-mediated mechanism for pathogen detection in intestinal epithelia.

STUDY QUESTION Could inhibition of the checkpoint kinase (CHEK) pathway protect human oocytes and even enhance the anti-tumour effects, during chemotherapy? SUMMARY ANSWER CHEK inhibitors prevented apoptosis of human oocytes induced by chemotherapy and even enhanced the anti-tumour effects. WHAT IS KNOWN ALREADY CHEK inhibitors showed ovarian protective effects in mice during chemotherapy, while their role in human oocytes is unclear. STUDY DESIGN, SIZE, DURATION This experimental study evaluated the ovarian reserve of young patients (120 patients) with cancer, exposed or not exposed to taxane and platinum (TP)-combined chemotherapy. Single RNA-sequencing analysis of human primordial oocytes from 10 patients was performed to explore the mechanism of oocyte apoptosis induced by TP chemotherapy. The damaging effects of paclitaxel (PTX) and cisplatin on human oocytes were also evaluated by culturing human ovaries in vitro. A new mouse model that combines human ovarian xenotransplantation and patient-derived tumour xenografts was developed to explore adjuvant therapies for ovarian protection. The mice were randomly allocated to four groups (10 mice for each group): control, cisplatin, cisplatin + CK1 (CHEK1 inhibitor, SCH 900776), and cisplatin + CK2 (CHEK2 inhibitor, BML277). PARTICIPANTS/MATERIALS, SETTING, METHODS In the prospective cohort study, human ovarian follicles were counted and serum AMH levels were evaluated. RNA-sequencing analysis was conducted, and staining for follicular damage (phosphorylated H2AX histone; γH2AX), terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labelling (TUNEL) assays and assessments of apoptotic biomarkers (western blot and immunofluorescence) were conducted in human ovaries. After the treatments, histological analysis was performed on human ovarian samples to investigate follicular populations, and oocyte damage was measured by γH2AX staining, BAX staining, and TUNEL assays. At the same time, the tumours were evaluated for volume, weight, and apoptosis levels. MAIN RESULTS AND THE ROLE OF CHANCE Patients who received TP chemotherapy showed decreased ovarian reserves. Single RNA-sequencing analysis of human primordial oocytes indicated that TP chemotherapy induced apoptosis of human primordial oocytes by causing CHEK-mediated TAp63α phosphorylation. In vitro culture of human ovaries showed greater damaging effects on oocytes after cisplatin treatment compared with that after PTX treatment. Using the new animal model, CHEK1/2 inhibitors prevented the apoptosis of human oocytes induced by cisplatin and even enhanced its anti-tumour effects. This protective effect appeared to be mediated by inhibiting DNA damage via the CHEK-TAp63α pathway and by generation of anti-apoptotic signals in the oocytes. LARGE SCALE DATA N/A. LIMITATIONS, REASONS FOR CAUTION This was a preclinical study performed with human ovarian samples, and clinical research is required for validation. WIDER IMPLICATIONS OF THE FINDINGS These findings highlight the therapeutic potential of CHEK1/2 inhibitors as a complementary strategy for preserving fertility in female cancer patients. STUDY FUNDING/COMPETING INTEREST(S) This work was financially supported by the National Natural Science Foundation of China (nos. 82001514 and 81902669) and the Fundamental Research Funds for the Central Universities (2021yjsCXCY087). The authors declare no conflict of interest.

Cesarean section (CS) confers increased risk of type I diabetes, asthma, inflammatory bowel disease, celiac disease, overweight and obesity, etc., in the offspring. However, the underlying mechanism remains unknown. To investigate the influence of CS on gene expression in cord blood, we have performed RNA-sequencing followed by single-gene analysis, gene set enrichment analysis, gene co-expression network analysis, and interacting genes/proteins analysis in eight full-term infants born by elective CS and eight matched vaginally delivered (VD) infants. Crucial genes identified above were further validated in another 20 CS and 20 VD infants. We found for the first time that mRNA expression of genes involved in immune response ( IL12A , INFG , IL1B , TNF , MIF , IL4 , CA1, IFI27, HLA-DOB and EPHB1 ) and metabolism ( DLK1 , CYP2A6 and GATM ) were significantly influenced by CS. Notably, serum TNF-α and IFN-γ were remarkably up-regulated in the CS infants ( p = 5.0 × 10 –4 and 3.0 × 10 –3 , respectively) compared to the VD infants. It is biologically plausible that CS may exert adverse impacts on offspring health through influencing expression of genes in the above processes. These findings will help understand the potential underlying mechanisms of the adverse health impacts of CS and identify biomarkers for future health of offspring born with different delivery modes.

Abstract Objective To look into the physiological functions of the lncRNA DLEU2 in the tumorigenesis of oral squamous cell carcinoma (OSCC), as well as whether it plays a role in the emergence and advancement of OSCC by governing SIX1. Methods The inhibitory role of DLEU2 on the proliferation of SCC-15 cells was examined by CCK8. Flow cytometry was used to study the influence of DLEU2 inhibitory activity on SCC-15 apoptotic cell death. In addition, trans-well assays were used to analyze the influence of DLEU2 suppression on SCC-15 cell differentiation and proliferation. Results The DLEU2 expression in OSCC cancerous specimens was considerably stronger than the corresponding healthy tissues; and DLEU2 was elevated in all four OSCC cells. The immunohistochemistry data also showed the level of DLEU2 was also greatly elevated in OSCC tissues than healthy specimens. After transfection of si-DLEU2, the viability of SCC-15 cells decreased significantly. Additionally, the number of apoptosis cells transfected with si-DLEU2 was significantly higher than controls. Using trans-well invasion assay, the data suggested the number of invasive cells formed by blocking DLEU2 of SCC-15 and SCC-25 cells was markerly lower than the controls. The results of ECAE and OCR also showed that DLEU2 could promote the glycolysis of OSCC cells while inhibit the oxidative phosphorylation progress of OSCC cells. Our subsequent analysis of the main enzymes affecting glycolysis, GLUT1 and HK2, showed that blocking expression of DLEU2 is able to obviously reduce the GLUT1 level, but not HK2. Subsequent ChIP experiments confirmed that SIX1 could bind to the promoter of GLUT1, and knocking down DLEU2 could reduce the binding ability of SIX1 to the promoter of GLUT1. Finally, we utilized luciferase assays to confirm that knockdown of DLEU2 expression could directly reduce GLUT1 transcript levels. The results of ECAR and OCR experiments also showed that overexpression of SIX1 could reverse the decreased glycolysis of OSCC cells brought down by knockdown of DLEU2. Conclusion DLEU2 is essential for OSCC tumorigenesis, migratory and glycogenolysis. The DLEU2/SIX1 role is implicated in OSCC cell invasion and aerobic glycolysis.

Cosmetics are an important aspect of the lives of many people. With an increasing demand for cosmetics, consumers pay more attention to their efficacy and composition. To improve their efficacy, prohibited substances, such as hormones, glucocorticoids, antibiotics, antifungals and antihistamines, may be added to cosmetics. We developed a rapid method for the multi-class analysis of drug residues in toner and lotion cosmetic samples using high-performance liquid chromatography coupled with quadrupole time-of-flight high-resolution mass spectrometry (HPLC-Q-TOF-HRMS). The primary variables in the extraction and purification steps were studied to minimize the interference of the sample matrix. The non-information-dependent sequential window acquisition of all theoretical fragment ion spectra (SWATH®) mode was used to improve the data acquisition efficiency. The secondary product ion peak areas were used for quantification to obtain a satisfactory matrix effects. The validation experiments confirmed that the developed method exhibited good linearity (5-200 ng/L) with correlation coefficients (R) ≥ 0.9902. Our developed method was then successfully applied to 92 real cosmetic samples. The calibration curve established by this method can be used for retrospective quantitative analysis over long durations without re-calibration. This method is efficient and suitable for screening and controlling multi-class prohibited substances in the cosmetics industry to reduce potential risks.

Semaphorins are a family of evolutionarily conserved morphogenetic molecules that were initially found to be associated with axonal guidance. Semaphorin 4C (Sema4C), a member of the fourth subfamily of semaphorins, has been demonstrated to play multifaceted and important roles in organ development, immune regulation, tumor growth, and metastasis. However, it is completely unknown whether Sema4C is involved in the regulation of ovarian function. We found that Sema4C was widely expressed in the stroma, follicles, and corpus luteum of mouse ovaries, and its expression was decreased at distinct foci in ovaries of mice of mid-to-advanced reproductive age. Inhibition of Sema4C by the ovarian intrabursal administration of recombinant adeno-associated virus-shRNA significantly reduced oestradiol, progesterone, and testosterone levels in vivo. Transcriptome sequencing analysis showed changes in pathways related to ovarian steroidogenesis and the actin cytoskeleton. Similarly, knockdown of Sema4C by siRNA interference in mouse primary ovarian granulosa cells or thecal interstitial cells significantly suppressed ovarian steroidogenesis and led to actin cytoskeleton disorganization. Importantly, the cytoskeleton-related pathway RHOA/ROCK1 was simultaneously inhibited after the downregulation of Sema4C. Furthermore, treatment with a ROCK1 agonist after siRNA interference stabilized the actin cytoskeleton and reversed the inhibitory effect on steroid hormones described above. In conclusion, Sema4C may play an important role in ovarian steroidogenesis through regulation of the actin cytoskeleton via the RHOA/ROCK1 signaling pathway. These findings shed new light on the identification of dominant factors involved in the endocrine physiology of female reproduction.