Original research publications

Exposure to mercury (Hg) is well-established to induce menstrual cycle disorders and disrupt sex hormone functions. Astragalus species, characterized by their potent antioxidant and anti-apoptotic properties, have shown potential in mitigating these adverse effects by promoting folliculogenesis and restoring hormonal balance. This study aimed to evaluate the protective effects of Astragalus microcephalus polysaccharides (APS), both alone and in combination with vitamin E and selenium (Vit E+Se), against ovarian damage and oxidative stress induced by mercuric chloride toxicity in rats. Thirty-six female Wistar rats were synchronized via vaginal smear and randomly assigned to six groups: Control, Mercury (0.43 mg/kg HgCl2), APS (2.7 g/kg), Vit E+Se, Mercury+APS and Mercury+Vit E+Se. Following a 15-day subacute exposure period, serum levels of estradiol, LH and FSH were measured. Oxidative stress markers and ovarian histopathology were evaluated. Apoptotic and inflammatory pathways were assessed through immunohistochemical staining of Caspase-3, AIF and NF-κB. Hg-induced reductions in hormone levels were significantly restored to near control levels in the APS-treated groups and antioxidant levels were notably elevated in these groups, indicating mitigation of oxidative stress. Furthermore, immunohistochemical analysis revealed a significant reduction in stromal and follicular degeneration and a decrease in apoptotic activity in the ovaries of rats treated with APS. APS exerts a significant protective effect against mercury-induced ovarian toxicity. This protection is mediated through the attenuation of oxidative stress, inflammation and apoptosis, as evidenced by the modulation of Caspase-3, AIF and NF-κB pathways. The results suggest that APS, particularly in synergy with vitamin E and selenium, offers a potential therapeutic strategy for managing heavy metal-induced reproductive damage.

Ambient heat is an escalating climate-related hazard, yet its implications for menstruation and adolescent and young adult mental health remain understudied. Menstruating individuals face unique physiological and psychosocial challenges that may be intensified in the presence of heat stress. These challenges can be further shaped by stigma, period poverty, and developmental vulnerability, with potential implications for depression, anxiety, and psychological wellbeing. Heat exposure may disrupt thermoregulation, exacerbate menstrual symptoms, and heighten psychological stress, but the combined effects of these processes remain poorly understood. In this critical narrative review, we: draw together evidence from multiple disciplines on biological, psychological, and structural pathways linking heat exposure, menstruation, and mental health. We identify important research gaps and propose a conceptual framework and gender-responsive research agenda to inform equitable climate-resilient health strategies.

This study investigated the association between dietary quality indices and female infertility using data from the National Health and Nutrition Examination Survey (NHANES) 2013-2020. A total of 5,477 women aged 18-59 years were included. Dietary quality was assessed using 24-hour dietary recall data and summarised by four predefined indices: the alternate Mediterranean diet score (MED), the Healthy Eating Index-2020 (HEI-2020), the Alternate Healthy Eating Index (AHEI), and the Dietary Approaches to Stop Hypertension (DASH) score. Weighted logistic regression models were used to estimate the associations between dietary quality and infertility, and restricted cubic spline (RCS) analyses were used to examine potential dose-response patterns. After adjustment for age, race/ethnicity, education level, body mass index (BMI), smoking status, alcohol consumption, marital status, menstrual regularity, age at menarche, history of pelvic inflammatory disease, and total energy intake, higher MED, HEI-2020, AHEI, and DASH scores were inversely associated with infertility. RCS analyses suggested approximately linear inverse associations for these indices (all P for nonlinearity >0.05). Higher overall diet quality was associated with lower odds of infertility among adult women in the United States (US). Because this was a cross-sectional study using self-reported infertility, the findings should be interpreted as associative rather than causal.

Seated whole-body vibration (WBV) exposure increases musculoskeletal injury (MSKI) risk, particularly for females. This review synthesizes the literature evidence for sex-differences in biodynamic response (BR) to seated WBV. Studies were included if they reported one or more BR outcomes, included both males and females, and exposed participants to seated WBV. Twenty-six studies met the inclusion criteria, fourteen of which reported significant sex-differences in one or more BR outcome(s) under various vibration and seating conditions. Experimental and data analysis methods between studies were found to be somewhat inconsistent and/or incompletely disclosed, making comparisons between studies difficult. This potentially led to the equivocal findings between studies with respect to sex differences. Most authors suggest that regional body composition probably contributes to differences in BR but no studies have assessed body composition by region.

Yiqi Huoxue Formula (YQHXF) is an 11-herb hospital-based traditional Chinese medicinal formula developed according to the therapeutic principle of tonifying qi and activating blood circulation. Previous clinical application has suggested its relevance to postoperative ovarian endometriosis with qi deficiency and blood stasis; however, its pharmacological basis remains incompletely understood. This study investigated whether YQHXF attenuates endometriotic lesion progression by modulating NF-κB p65/NLRP3 inflammasome-associated pyroptotic signaling. A rat model of endometriosis was established by autologous endometrial transplantation. LC-HRMS was used to characterize the chemical profile of YQHXF, and RNA sequencing was performed on ectopic lesions. Primary eutopic and ectopic endometrial stromal cells (ESCs) were isolated from patients with endometriosis. p65 overexpression, siRNA-mediated p65 knockdown, the NLRP3 inhibitor MCC950, and the NLRP3 agonist BMS-986299 were used to evaluate pathway involvement. Cell viability, inflammatory cytokines, inflammasome- and pyroptosis-associated markers, transmission electron microscopy, PI/Hoechst staining, and LDH release were assessed. LC-HRMS profiling yielded 287 putatively annotated records and showed recurrent chemical features across three independent YQHXF batches. YQHXF reduced ectopic lesion volume, alleviated histopathological injury, and downregulated Ki-67 and PCNA in vivo. Transcriptomic analysis linked its effects to immune-inflammatory responses, TNF/NF-κB signaling, NOD-like receptor signaling, and cytoskeletal/adhesion remodeling. In EM tissues and ectopic ESCs, NF-κB p65/NLRP3 inflammasome-associated signaling was activated, accompanied by increased IL-1β and IL-18 release, elevated cleaved caspase-1 and GSDMD-N, reduced E-cadherin, and pyroptosis-associated membrane injury. YQHXF suppressed these changes, whereas NLRP3 activation or p65 overexpression partially weakened its protective effects. YQHXF attenuates endometriosis progression, at least in part, by suppressing NF-κB p65/NLRP3 inflammasome-associated pyroptotic signaling and inflammatory injury. These findings provide preclinical mechanistic evidence supporting further investigation of YQHXF as a potential non-hormonal therapeutic candidate for endometriosis.

Polycystic ovary syndrome (PCOS) is a prevalent endocrine disorder characterized by ovulatory dysfunction, hyperandrogenism, and insulin resistance. It results from complex genetic and environmental factors and progressively impairs reproductive and metabolic health. Natural phytotherapy provides holistic regulation through multiple components and targets and is recognized for its favorable safety profile, including low organ toxicity. Consequently, it represents a promising avenue for comprehensive PCOS management. This review aims to comprehensively analyze the ethnopharmacological applications of natural herbs in modulating ferroptosis within the context of PCOS. This study systematically reviewed natural herbal interventions for PCOS, with a focus on their mechanisms in regulating ferroptosis. It consulted traditional medical classics, ethnomedicinal records, and clinical literature, and searched databases including PubMed and the China National Knowledge Infrastructure (CNKI). Keywords such as "PCOS," "ferroptosis," "natural herbal medicines," "traditional Chinese medicine (TCM) compound preparations," "single-herb extracts," and "plant active ingredients" were used to retrieve research from ethnobotany, phytochemistry, and pharmacology. Recent relevant publications were comprehensively collected and analyzed. Integrating natural herbal medicines into the comprehensive management of PCOS offers several advantages, which can be primarily explained from two perspectives. First, the pathological mechanisms of PCOS involve multiple interconnected biological processes, including hormonal imbalance, insulin resistance, oxidative stress, and chronic inflammation. The onset and progression of PCOS are closely linked to cell death pathways such as ferroptosis and are regulated by various molecular pathways, including GPX4/COX2, SLC7A11/GPX4, circ_0097636/miR-186-5p/SIRT3, and AMPK/Nrf2 signaling. Drugs targeting a single molecule often struggle to effectively regulate such a complex network. In contrast, various herbal medicines and their active components-such as platycodin D, nuciferine (NF), baicalein, and berberine (BBR)-exhibit multi-target properties, enabling them to simultaneously modulate key molecules involved in ferroptosis in PCOS, including GPX4, SLC7A11, COX2, FTH1, ACSL4, and ferritin. This allows for synergistic intervention in multiple pathological processes of the disease. Additionally, natural herbal medicines and their active components have demonstrated a high level of safety with long-term use, particularly exhibiting relatively lower risks of liver and kidney toxicity. They offer certain advantages compared to traditional hormones or insulin sensitizers. For example, classic compound prescriptions such as Wenshen Tiaojing decoction (WSTJD), Shoutai pill, and Erchen decoction are commonly used in TCM clinical practice to regulate menstruation, improve insulin resistance, and promote ovulation, supported by extensive practical evidence. In recent years, to clarify their scientific basis, an increasing number of studies have extracted key active components from these effective compound formulations using modern separation techniques. With the aid of cell and animal experiments, researchers have thoroughly elucidated the mechanisms by which these components improve PCOS through pathways such as the regulation of ferroptosis, thereby bridging traditional knowledge and modern pharmacology. Traditional Chinese compound formulations hold significant promise for developing drugs targeting PCOS. Notably, certain prescriptions and herbal extracts that focus on regulating ferroptosis as a key mechanism are attracting increasing attention and research within the academic community.

Early diagnosis of ovarian cancer remains a major clinical challenge. Human epididymis protein 4 (HE4) has emerged as a promising biomarker for early OC detection. To address limitations of existing HE4 quantification methods, we developed a nano-optical biosensor based on a luminescent europium-thiosemicarbazide (Eu-TSC) complex immobilized within a modified epoxy-cellulose thin-film matrix. The Eu-TSC thin film exhibited characteristic europium excitation and emission bands at 395 nm and 617 nm. Following functionalization with a monoclonal anti-HE4 antibody, the biosensor displayed a concentration-dependent fluorescence quenching response upon HE4 binding. A linear analytical response was obtained over 0.05-0.37 ng mL -1 , with LOD of 0.108 ng mL -1 . However, the LOQ (0.327 ng mL -1 ) lies close to the upper limit of the linear range, the practical validated quantification interval is extremely narrow (approximately 0.327-0.37 ng mL -1 ). This interval is far too narrow to cover the clinical HE4 range, and the present sensor should therefore be regarded as a preliminary proof-of-concept platform rather than a clinically suitable quantitative assay. Time-resolved luminescence measurements are consistent with a quenching-based sensing response, but the present experiments do not distinguish between FRET, PET, or other quenching pathways. Molecular docking and 50 ns molecular dynamics simulations provide a computational rationale for ligand selection and suggest possible surface binding of thiosemicarbazide-based ligands to HE4. Selectivity studies against twelve potential interferents revealed relative responses below 5% for all tested analytes when tested individually at clinically relevant concentrations (n = 3 per interferent; one-way ANOVA with Dunnett's test). However, because interferents were tested individually rather than as pooled mixtures, and because cross-reactivity with structurally related proteins cannot be fully excluded, these selectivity results are preliminary and do not establish excellent selectivity in complex serum. The biosensor was further evaluated using 21 human serum samples (11 OC patients, 10 healthy controls), demonstrating agreement with a commercial HE4 ELISA assay (R 2  > 0.98). Recovery values were re-verified for all samples; several values fall outside the commonly accepted 85-115% range and are flagged in Tables 1A and B. Therefore, the analytical accuracy of the present method is preliminary and not satisfactory for clinical quantification. ROC analysis yielded an AUC of 1.000, with 90.9% sensitivity and 100% specificity. These findings are based on a limited case-control cohort (n = 21) and must be interpreted as a preliminary proof-of-concept analytical and clinical evaluation, not as a clinically validated diagnostic platform. Larger prospective, multi-center studies including patients with benign gynecological conditions are required before clinical implementation can be considered.

Whether perineal injury severity independently determines postpartum sexual quality of life (SQoL) is unclear. This study evaluated perineal tears and body image as determinants of SQoL and sexual resumption timing. A total of 173 women (2-24 months postpartum, vaginal delivery) completed an online questionnaire including the SQoL-F and Body Image Scale (BIS). Women with perineal tear/episiotomy (PT/E; n = 118) were compared with those with an intact perineum ( n = 51) using multivariate regression. SQoL-F scores were similar between groups (71.1 &#xb1; 23.3 vs 76.9 &#xb1; 24.0; p = 0.10), though PT/E delayed sexual resumption ( p = 0.04). On multivariate analysis, SQoL-F was independently associated with dyspareunia (&#x3b2; = -10.96; p < 0.001), body image disturbance (&#x3b2; = -0.93; p < 0.001), and pre-pregnancy sexual satisfaction (&#x3b2; = +4.99; p = 0.001), but not tear type. Delayed resumption was independently associated with OASI (OR = 7.02; p = 0.02), first/second-degree tears (OR = 2.20; p = 0.04), and lower pre-pregnancy relationship satisfaction (OR = 0.50; p = 0.048). Postpartum SQoL is primarily determined by dyspareunia and body image disturbance rather than perineal tear severity. OASI carries a substantial risk of delayed sexual resumption, warranting dedicated postpartum counseling.

To explore how professional women undergoing assisted reproductive technology (ART) experience and manage conflicts between treatment demands and professional identity; and to identify ways healthcare professionals can support them. Following COREQ guidelines, semi-structured, in-depth interviews were conducted with 18 professional women undergoing ART in a tertiary care hospital. Reflexive Thematic Analysis bolstered by interdisciplinary investigator triangulation (clinical and basic science) ensured thematic confirmability. Analysis revealed Gendered Occupational Asymmetry (female occupational burden regardless of clinical aetiology), the Insider Paradox (the role reversal for healthcare professionals) and the Workplace Panopticon (constant self-surveillance to preserve professional authority). Interestingly, work simultaneously often acted as a Sanctuary of Competence , providing psychological respite. Finally, narratives exposed an Existential Rumination (mourning past personal-professional decisions) and permanent temporal career stalling. These themes were further mapped onto a 'A Day in the Life' structural framework to illustrate compounding daily burdens. Current clinical lifestyle directives inadequately account for the rigid psycho-socioeconomic structures governing professional women. Based on identified themes, we propose targeted measures for healthcare professionals to deliver patient-centric care by integrating occupational awareness and identity-sensitive counselling into routine ART practice, thereby potentially reducing distress and improving treatment experience.

Genital warts caused by human papillomavirus are common sexually transmitted infections, with no single therapy providing consistent clearance. Garlic extract has antiviral and antiproliferative properties and may offer a low-cost treatment. To compare the efficacy and safety of topical 5% garlic extract with 20% podophyllin solution in women with genital warts. Seventy-four women were randomized to 5% garlic extract ( n &#x2009;=&#x2009;37) or 20% podophyllin ( n &#x2009;=&#x2009;37), applied weekly for four weeks. Wart size and count were assessed using marginal models (generalized estimating equations). The primary outcome was change in mean wart size and the secondary outcome was change in wart count. Mean wart size decreased from 1.61&#x2009;&#xb1;&#x2009;0.81 to 0.60&#x2009;&#xb1;&#x2009;0.82&#x2009;mm with garlic extract and from 2.96&#x2009;&#xb1;&#x2009;7.99 to 0.62&#x2009;&#xb1;&#x2009;0.73&#x2009;mm with podophyllin. The between-group p -value at final assessment was 0.756; group&#x2009;&#xd7;&#x2009;time interaction p values were 0.329 for wart size and 0.583 for count. Among 35 and 37 participants analyzed, complete response occurred in 18 (51.4%) and 19 (51.4%), respectively; complete or partial response occurred in 23 (65.7%) and 25 (67.6%). Itching and burning occurred in 8 (21.6%) garlic-treated participants each. Both treatments reduced wart size and count, with similar response rates. Four-week follow-up precluded assessment of durability and recurrence.

Magnetic hyperthermia (&#x2264;45&#x202f;&#xb0;C) is a promising strategy for conditions requiring localized tissue disruption in sensitive anatomical regions, including ectopic pregnancy (EP), where temperature-sensitive trophoblasts can be selectively disrupted by modest heating. However, effective treatment requires nanoparticles that rapidly reach therapeutic temperatures after systemic administration while attenuating heat generation as the target temperature is approached to minimize damage to surrounding tissues. Here, cobalt-doped soft ferromagnetic iron oxide nanoparticles (Co-FIONs) were engineered to achieve this balance through classic temperature-dependent coercivity within the hyperthermia window. Coercivity decreased 2.27-fold at 45&#x202f;&#xb0;C, accompanied by a 1.58-fold reduction in heating rate from 1.9 to &#x223c;1.2&#x202f;&#xb0;C&#x202f;s -1 , progressively attenuating heat generation while maintaining a high specific absorption rate (SAR) of 2907&#x202f;&#xb1;&#x202f;84&#x202f;W&#x202f;g -1 Fe. Box-Lucas analysis further demonstrated progressive temperature-dependent attenuation of heating kinetics, consistent with the combined effects of magnetic softening and thermal dissipation. Under biologically relevant conditions, Co-FIONs@PP retained a high SAR of 474&#x202f;&#xb1;&#x202f;56&#x202f;W&#x202f;g -1 Fe in 2.5% agar, demonstrating its ability to achieve therapeutically relevant magnetic hyperthermia at clinically feasible iron doses. Systemically administered Co-FIONs@PP also provided T 2 /T 2 *-weighted MRI contrast for localization of implantation sites. In pregnant mice, Co-FIONs@PP accumulated at fetoplacental units and, following two alternating magnetic field exposures, induced localized trophoblast apoptosis and selective gestational sac disruption while preserving adjacent endometrium and subsequent fertility. These findings establish Co-FIONs@PP as a promising theranostic platform for MRI visualization and localized magnetic hyperthermia, providing a foundation for minimally invasive, image-assisted treatment of EP.

Cigarette smoking and nicotine exposure promote breast cancer progression and poor outcomes by aberrantly activating nicotinic acetylcholine receptors (nAChRs), with &#x3b1;9-nAChR serving as a key oncogenic driver, especially in triple-negative breast cancer (TNBC). However, effective therapeutic strategies that directly target &#x3b1;9-nAChR remain limited. In this study, we engineered an antigen comprising the extracellular loops of &#x3b1;9-nAChR (&#x3b1;9CLP) and used mouse immunization followed by phage display to isolate an &#x3b1;9CLP-specific single-chain variable fragment (scFv) antibody, mCLPS1. The dominant clone mCLPS1 showed specific binding to &#x3b1;9-nAChR and was further engineered into an Fc-fusion antibody (mCLPS1-Fc) and an anti-mPEG bi-specific antibody (mCLPS1/anti-mPEG BsAb) to improve its affinity and therapeutic potential. The mCLPS1/anti-mPEG BsAb significantly enhanced the therapeutic efficacy of mPEGylated liposomal doxorubicin (LD) in targeting &#x3b1;9-nAChR-positive breast cancer. In TNBC patient-derived xenograft (PDX) models, mCLPS1-Fc markedly inhibited tumor growth, mitigated nicotine-induced tumor acceleration, and enhanced docetaxel's antitumor efficacy. Preliminary tolerability assessments of mCLPS1/anti-mPEG BsAb and mCLPS1 Fc antibody administration in non-human primates, cynomolgus macaque (Macaca fascicularis). did not reveal neuromuscular toxicity, myasthenia gravis-like symptoms, hematologic abnormalities, or overt organ injury within the observation period, supporting a favorable short-term tolerability profile under the tested conditions. In summary, this study identifies &#x3b1;9-nAChR as a key molecular target in nicotine-induced tumor progression and highlights &#x3b1;9-nAChR-specific antibodies as promising targeted therapeutic candidates for TNBC, particularly in patients with a history of nicotine exposure.

A major challenge for injectable therapeutic nanorobots is to achieve efficient and spatially precise intracellular actuation under biologically compatible field conditions. Here we introduce homotypic membrane-camouflaged magnetic nanorobots that couple selective tumor-cell internalization, low-field intracellular rotation and glutathione-responsive drug release for chemo-mechanical breast cancer therapy. The nanorobots comprise disulfide-bridged mesoporous organosilica nanohelices loaded with doxorubicin, decorated with Fe 3 O 4 nanoparticles and cloaked with MCF-7 cancer cell membranes. The membrane cloak promotes homologous recognition and enriches magnetic actuators inside MCF-7&#x202f;cells, where a rotating magnetic field drives rapid intracellular rotation and localized mechanical perturbation. Intracellular glutathione concurrently cleaves the disulfide-containing organosilica framework, triggering nanorobot degradation and doxorubicin release. This spatial coupling of mechanical injury and chemotherapy induces pronounced tumor-cell death in vitro and suppresses tumor growth by 87.03% in an MCF-7 xenograft model without obvious systemic toxicity. These findings establish homotypic intracellular localization as a strategy for programmable magneto-mechanical nanotherapy.

Nanoparticles (NPs) can be engineered to achieve targeted delivery with strategies based on surface modifications. These include layer-by-layer (LbL) NPs, modular electrostatically assembled carriers with tunable surface properties altered by changes to the outer polyelectrolyte layer. Variations in these polymers dictate intracellular trafficking and biodistribution patterns. As NPs are administered, a layer of protein adsorbs to their surfaces, forming a protein corona that affects NP properties, alters biodistribution, and ultimately impacts therapeutic efficacy. We hypothesized that some differences in LbL NP performance are due, in part, to variations in the resulting protein coronas. To study them, we first optimized an ultrafiltration method to effectively isolate LbL NPs with their protein corona. Following incubation in conditioned media, anionic homopolypeptide outer layers, such as poly-l-aspartic acid (PLD) and poly-l-glutamic acid (PLE), and LbL NPs with the bioinert polymer poly(acrylic acid) (PAA) had the lowest amount of protein associated, lower than conventional PEG liposomes. While mass spectroscopy revealed changes in the protein composition among LbL NPs; albumin, alpha-2-macroglobulin, and apolipoprotein B were most abundant, with PLE and HA NPs presenting unique protein profiles. In vitro, pre-formed protein coronas reduced uptake in macrophages but increased uptake in ovarian cancer cells for certain LbL NPs, which were protein-dependent. In vivo, LbL NP outer layer influenced both serum half-life and biodistribution. Overall, this work highlights that LbL NPs can be designed to control protein corona formation and supports the idea that further understanding NP interactions with biological fluids is essential for designing clinically translatable NP platforms.

Physiological human chorionic gonadotropin (hCG) elevations in postmenopausal women are prone to misclassification as pathological. Substantial inter-system variation exists among immunoassays, and system-specific reference intervals (RIs) for this population have not been established. This study aimed to establish 97.5th percentile RIs and 99th percentile thresholds for serum hCG in healthy postmenopausal women across four chemiluminescence immunoassay (CLIA) systems. A total of 460 postmenopausal women were enrolled. Serum hCG was measured using four immunoassay systems: Beckman UniCel DxI 800 (A), Siemens Atellica IM 1600 (B), Roche Cobas e 801 (C), and Abbott ARCHITECT i2000SR (D). Analytical precision was evaluated according to Clinical and Laboratory Standards Institute (CLSI) EP15-A2. Samples with hCG&#xa0;>&#xa0;5.0&#xa0;IU/L (> 6.0&#xa0;IU/L for System B) underwent polyethylene glycol (PEG) precipitation to exclude macro-complex interference. Spearman rank correlation was used to assess correlations between hCG and age, follicle-stimulating hormone (FSH), and luteinizing hormone (LH). One-sided upper reference limits (97.5th and 99th percentiles) were estimated using the non-parametric method. 355 subjects were included in the final analysis. The imprecision of all four systems met the analytical performance criterion. Serum hCG showed poor positive correlations with age (Spearman r&#xa0;=&#xa0;0.146-0.198, all P&#xa0;<&#xa0;0.01). Fair correlations were observed between hCG and FSH (r&#xa0;=&#xa0;0.341, 95% CI: 0.246-0.430) and between hCG and LH (r&#xa0;=&#xa0;0.428, 95% CI: 0.339-0.509). The 97.5th percentile RIs were&#xa0;<&#xa0;7.1, < 8.2, < 5.7, and&#xa0;<&#xa0;5.9&#xa0;IU/L for Systems A-D, respectively; the 99th percentile thresholds were&#xa0;<&#xa0;8.8, < 9.3, < 7.1, and&#xa0;<&#xa0;7.8&#xa0;IU/L. System B consistently yielded the highest thresholds, while System C produced the lowest. The established 97.5th percentile RIs and 99th percentile thresholds provide reliable, system-specific quantitative criteria for differentiating pituitary-derived from pathological hCG elevations.

Breast cancer (BC) is the leading cause of cancer-related mortality among women worldwide. Early, non-invasive, and accurate detection remains critical to reducing mortality and improving clinical outcomes. Volatilomics, focused on the characterization of volatile organic compounds (VOCs) emitted by the human body, offers a promising approach for disease diagnosis. VOC-based testing has already been translated into routine clinical practice for a number of illnesses, yet such tests for BC remain unavailable. To accelerate translational progress within this field, a comprehensive synthesis of available evidence is urgently required. Existing reviews on volatilomics in BC are either constrained to single matrices (e.g., exhaled breath), focus on exogenous VOCs, provide only cursory coverage of BC within multi-cancer overviews, or fail to reflect recent advances. In this narrative review, we synthesize evidence from 46 studies published over the past two decades, including studies of exhaled breath (n&#x202f;=&#x202f;24), urine (n&#x202f;=&#x202f;10), exhaled breath condensate (EBC, n&#x202f;=&#x202f;1), sweat (n&#x202f;=&#x202f;2), tumor tissue (n&#x202f;=&#x202f;1), cell lines (n&#x202f;=&#x202f;6), and animal tumor models (n&#x202f;=&#x202f;2). We review the application of volatilomics in diagnostic potential, pathological and molecular subtyping, clinical staging, risk prediction, and VOC generation mechanisms. Furthermore, we evaluate the impact of advanced analytical methods, such as the integration of volatilomics with other omics approaches to enhance diagnostic performance. In addition, we outline the major translational barriers restricting clinical transformation, followed by an in-depth discussion of prospective research directions including multi-matrix data integration, AI-assisted volatilomic modeling, and living olfactory biosensors. This review delivers an integrated framework to support subsequent mechanistic exploration and the transition from analytical feasibility toward clinical validation of VOC-based BC detection.

Surface-enhanced Raman spectroscopy (SERS) has emerged as a powerful analytical platform for addressing the critical challenges in breast cancer precision diagnostics. This review systematically examines the recent methodological advances that position SERS as a transformative tool across the entire clinical continuum of breast cancer management-spanning non-invasive liquid biopsy screening, intraoperative surgical margin navigation, and personalized therapeutic guidance-with an emphasis on analytical innovation rather than clinical outcomes alone. We critically assess the core analytical strategies that underpin these applications, including the rational design of high-performance plasmonic nanostructures for enhanced signal reproducibility and amplification; the development of targeted and label-free SERS probes for ultrasensitive detection of circulating biomarkers such as microRNAs, exosomal proteins, and circulating tumor cells in complex biofluids; the integration of SERS with microfluidic platforms to enable sample-in-answer-out automation with reduced sample volumes and accelerated turnaround times; and the application of artificial intelligence and deep learning algorithms for deciphering complex spectral "fingerprints" enabling automated disease classification and molecular subtyping. Furthermore, we discuss the persistent analytical bottlenecks hindering clinical translation, notably the lack of standardized substrate fabrication protocols, signal variability due to heterogeneous biological matrices, and the biosafety evaluation of nanoprobes. By highlighting these methodological advancements and unresolved analytical challenges, this review aims to provide a comprehensive resource for analytical chemists and bioengineers seeking to develop next-generation SERS-based diagnostic platforms, ultimately contributing to the evolution of breast cancer management toward more precise, data-driven, and personalized paradigms.

In pursuit of non-invasive strategies for early cancer diagnosis and improved global health outcomes, we report a sensitive electrochemical sensor based on Gd-doped NiAl-LDH integrated with f-MWCNT for detecting adenosine, a biomarker associated with breast cancer progression. The exponential doping of Gd in NiAl-LDH leads to distortion, as evidenced by a shift towards lower 2&#x3b8; in XRD, progressively increased lattice spacing in HR-TEM, and a shift towards lower binding energy in XPS. The 1% Gd-NiAl LDH/f-MWCNT composite on SPCE comprises multiple electroactive sites that facilitate electron transfer, yielding robust oxidation performance and responsiveness. A low detection limit of 13&#x202f;nM with a linear range of (0.05- 67.7&#x202f;&#x3bc;M) and great stability was achieved with the 1% Gd-NiAl LDH/f-MWCNT sensor in DPV. The proposed sensing platform demonstrated promising analytical performance on pretreated biological samples with a miniaturized, portable Zensor potentiostat, highlighting its potential as a laboratory-scale proof of concept for future point-of-care diagnostic applications. This platform may pave the way for developing point-of-care diagnostics in the early stages and localized therapy.

Human epididymis protein 4 (HE4) serves as a crucial biomarker for the early diagnosis and monitoring of ovarian cancer. Herein, a novel trimetallic metal-organic framework (CoFeZn-MOF) was engineered as a peroxidase-mimicking nanozyme, and its application in the development of a sandwich-type electrochemical immunosensor for the highly sensitive detection of HE4 was demonstrated. Compared to single-metal or bimetallic MOFs, CoFeZn-MOF nanozyme exhibited superior peroxidase-like activity, capable of catalyzing the decomposition of hydrogen peroxide while also efficiently oxidizing 3,3',5,5'-tetramethylbenzidine (TMB) to produce the blue oxTMB product. The process of hydrogen peroxide decomposition catalyzed by CoFeZn-MOF involved redox reactions that induced significant electron transfer. Consequently, detectable changes in reduction current occurred within the current-time curve, enabling signal amplification for quantitative analysis of the tumor marker HE4. Under the optimized conditions, the linear detection range of the immunosensor was 0.01-800&#x202f;ng/mL, with a low detection limit of 4.6&#x202f;pg/mL. In addition, the biosensor exhibited outstanding stability, reproducibility, and interference resistance. Furthermore, through the analysis of actual samples, the constructed biosensor had great potential in clinical applications, providing a novel high-sensitivity analytical tool for tumor marker detection.

The Xingxiao Pill (XXP), a representative traditional Chinese medicine (TCM) formula used to promote blood circulation and remove blood stasis in tumor therapy, has shown promising clinical potential. According to the book Famous Traditional Chinese Medicine Formulas and Case Studies for Cancer Treatment, XXP has demonstrated favorable efficacy in the treatment of breast cancer. The therapeutic effects and mechanisms of action of XXP in breast cancer have not been reported. The aim of the study was to evaluate the therapeutic efficacy of XXP in breast cancer and to investigate its potential mechanisms of action and active components. First, the proliferation-inhibiting activity of XXP against eight types of breast cancer cell lines was evaluated using the CCK-8 assay. Subsequently, the effects of XXP on the biological behavior of 4T1 cells were further investigated using cloning assays, EDU assays, and flow cytometric apoptosis assays. In addition, the mechanism of apoptosis was examined by western blotting (WB). A 4T1 subcutaneous tumor model was established to evaluate the in vivo antitumor activity of XXP. Concurrently, the safety of XXP was assessed through H&E staining of tissue specimens from major organs and by determining serum biochemical markers of liver and kidney function. Furthermore, weighted gene co-expression network analysis (WGCNA) was used to integrate metabolomics and transcriptomics data, to elucidate the mechanisms responsible for the anti-breast cancer activity of XXP. These findings were validated through WB experiments. Finally, bioactive components of XXP were identified and screened using UPLC-Q-TOF-MS/MS (UPLC-MS) and the SwissADME database, and their activity was validated using CCK-8 assays. Bioactive components with potential binding to mTOR protein were screened and identified using Biolayer Interferometry (BLI) combined with UPLC-MS technology and further confirmed through affinity experiments. XXP exhibited proliferation-inhibiting activity against various subtypes of breast cancer cells. In further studies, XXP demonstrated both proliferation-inhibiting and apoptosis-promoting activities against 4T1 breast cancer cells in vitro and in vivo. XXP induced mitochondrial apoptosis by activating the cytochrome C/caspase-9/caspase-3/PARP signaling cascade. Subsequently, integrated metabolomic and transcriptomic analyses revealed that XXP reshaped amino acid metabolism. WGCNA also identified a gene module highly correlated with differential changes in amino acid metabolites, which was enriched in the ErbB3/PI3K/AKT/mTOR signaling pathway. WB analysis revealed reduced levels of ErbB3 and PI3K proteins, as well as decreased phosphorylation levels of AKT/mTOR, although total AKT/mTOR levels remained unchanged. Consequently, we hypothesized that XXP regulates amino acid metabolism and promotes apoptosis by inhibiting the ErbB3/PI3K/AKT/mTOR signaling pathway. Finally, seven bioactive components of XXP were identified by UPLC-MS analysis and screening against the SwissADME database. Among these, the inhibitory activity of cinnamic acid, palmitic acid, eugenol, 11-keto-&#x3b2;-boswellic acid, and asiatic acid against 4T1 breast cancer cell proliferation was validated using CCK-8 assays. Furthermore, cinnamic and palmitic acid were identified as active components that may bind to mTOR. XXP exhibits pharmacological activity that inhibits 4T1 breast cancer cell proliferation and promotes apoptosis in vitro and in vivo. XXP may exert these effects by inhibiting the ErbB3/PI3K/AKT/mTOR signaling pathway, regulating amino acid metabolism, and promoting caspase-mediated endogenous apoptosis. Cinnamic acid, palmitic acid, eugenol, 11-keto-&#x3b2;-boswellic acid, and asiatic acid were identified and validated as active compounds in XXP. Among these, cinnamic acid and palmitic acid are active compounds that potentially bind to mTOR. This study provides experimental data supporting the use of XXP for the treatment of breast cancer.

Mitochondria are emerging as critical hubs for cancer therapeutics, yet the development of agents that simultaneously enable real-time imaging and targeted therapy remains a formidable challenge. Here we report LN1, a smart theranostic small molecule designed to selectively target mitochondrial G-quadruplex (mtG4) DNA through a triphenylphosphonium (TPP) moiety integrated with a triphenylamine-derived fluorescent scaffold. LN1 exhibits high-affinity binding to mtG4 structures (K d &#xa0;=&#xa0;0.2&#xa0;&#x3bc;M) with a 1:2 stoichiometry, leading to a pronounced fluorescence turn-on response via restriction of intramolecular rotation. This optical property enables real-time, subcellular tracking of LN1 localization to mitochondria in live cancer cells with exceptional specificity. Functionally, LN1 stabilizes mtG4s, disrupts mitochondrial homeostasis, elevates reactive oxygen species, depletes ATP, and induces S-phase arrest, ultimately triggering robust apoptosis through activation of the caspase-3/7/9 and PARP pathways. In vivo, LN1 achieves selective tumor accumulation, enables high-contrast fluorescence imaging of xenografted tumors, and exhibits antitumor efficacy comparable to that of topotecan. By integrating diagnostic imaging and targeted anticancer activity within a single molecular framework, this work establishes a new paradigm for mitochondria-targeted theranostics and highlights mtG4 DNA as a clinically actionable vulnerability in cancer.

Gestational diabetes mellitus (GDM) is associated with adverse maternal and neonatal outcomes, but molecular alterations across the maternal-placental-fetal interface remain incompletely defined. We applied data-independent acquisition mass spectrometry to matched maternal blood (MB), umbilical cord blood (UCB) and placental tissue collected at delivery from 28 women with GDM and 21 controls. Differential protein abundance analysis, Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analysis, gene set enrichment analysis, weighted co-expression network analysis and pathway-level corroboration using two independent public placental transcriptomic cohorts were performed. Proteomic changes were compartment specific. Placental tissue showed the greatest number of differentially abundant proteins, whereas maternal blood and umbilical cord blood showed more limited alterations. Exploratory pathway-level analyses in MB suggested metabolic stress- and extracellular matrix-related patterns, whereas UCB suggested immune- and coagulation-related patterns. Placental tissue showed more prominent alterations involving extracellular matrix remodeling and ribosome-related pathways. A GDM-associated placental co-expression module contained both extracellular matrix proteins and ribosomal proteins. Independent transcriptomic datasets provided additional pathway-level support for ribosome-related and inflammatory processes. These findings suggest that GDM is associated with distinct proteomic changes in maternal blood, umbilical cord blood and placental tissue at delivery, with the most prominent alterations involving placental extracellular matrix remodeling and ribosome-related processes. SIGNIFICANCE: This matched multi-compartment proteomic study provides a placenta-centered view of GDM-associated molecular alterations at delivery. By integrating DIA proteomics, co-expression network analysis and pathway-level corroboration using independent transcriptomic datasets, the study identifies placental extracellular matrix remodeling and ribosome-related processes, while exploratory analyses of UCB suggest immune- and coagulation-related pathway patterns. These findings provide a basis for future hypothesis-driven studies of GDM-related placental dysfunction.

Protein profiles of amniotic fluid extracellular vesicle-enriched preparations may provide information on the fetal and gestational intrauterine environment, but their normal variation remains incompletely characterized. We used data-independent acquisition-based quantitative proteomics to compare AF-EV-enriched preparations sampled at approximately 19 and 24&#xa0;weeks from pregnancies with normal fetal cytogenetic findings and uncomplicated outcomes. Of 3321 quantified proteins, 2611 detected in at least three of four samples in each group were included in the primary analysis without imputation. Primary analysis identified 904 candidates. Of these, 698 were also supported by the KNN sensitivity analysis with concordant direction, including 385 higher at 19&#xa0;W and 313 higher at 24&#xa0;W. A separate complete-case analysis identified 596 candidates, all within the 698-protein set. Nominal over-representation analysis using the 2611 eligible protein groups as the background highlighted metabolic/digestion and apical/brush-border annotations among 19&#xa0;W-higher candidates and proteasome-centered annotations among 24&#xa0;W-higher candidates. These enrichment findings are exploratory because the displayed P values were not adjusted for multiple comparisons. Given the small cohort and absence of independent validation and comprehensive EV characterization, the results are hypothesis-generating and cannot establish EV-specific gestational remodeling. SIGNIFICANCE: Gestational age is a recognized source of variation in total amniotic fluid, but its influence on proteins recovered from AF-EV-enriched preparations during mid-pregnancy is less well defined. Using DIA proteomics, we compared preparations sampled at approximately 19 and 24&#xa0;weeks from pregnancies with normal fetal cytogenetic findings and uncomplicated outcomes. Of 3321 quantified proteins, 698 were supported by both the primary analysis and KNN sensitivity analysis and were retained as hypothesis-generating candidates. Nominal over-representation analysis using the 2611 protein groups eligible for the primary analysis as the background highlighted metabolic/digestion and apical/brush-border annotations among 19&#xa0;W-higher candidates and proteasome-centered annotations among 24&#xa0;W-higher candidates. These unadjusted enrichment results are descriptive and do not establish EV-specific remodeling. The findings nevertheless indicate that gestational age should be considered when designing and interpreting AF-EV proteomic studies and provide candidates for validation in larger independent cohorts.

Ductal carcinoma in situ (DCIS) is the precursor of infiltrating ductal carcinoma (IDC); however, mechanisms underlying the invasive program of breast tumor epithelial cells remain largely unknown. Different assays and cell models have been developed to evaluate the invasive capacities of breast tumor cells (spheroids, organoids, tumor explants&#x2026;). These models recapitulate basic interactions between the neoplastic cells and their microenvironment in a simplified setup. Whenever possible, hypotheses and mechanisms should be validated based on the observation of tumor cells in their native environment in the mammary gland using animal models. Here, we describe specific tools to study early breast tumor progression, taking into consideration reducing the number of xenografted animals and having the capacity to follow tumor progression over time. Combinations of these approaches have revealed new and exciting insights as to explain how DCIS progresses into a more aggressive invasive lesion. This chapter details technical procedures to generating in situ tumors after intraductal inoculation, visualizing the tumor xenograft directly in the mammary gland of the injected mouse using a mammary window for intravital multi-photon imaging of tumor cells in their environment. Finally, we illustrate how this procedure can be useful to analyze morphological changes in the matrix microenvironment during early breast cancer progression.

HPV-associated cervical cancer remains a leading cause of cancer-related death in women worldwide. The role of bone marrow mesenchymal stem cells (BMSCs) and miR-129 in tumor progression remains unclear. To investigate miR-129 expression and its clinical significance in HPV-associated cervical cancer and to explore whether BMSCs regulate malignant phenotypes through miR-129. miR-129 expression was measured by qRT-PCR in 77 cervical cancer and 30 normal cervical tissues. Associations with clinicopathological features and overall survival (OS) were analyzed. Diagnostic value was assessed by ROC curve. Rat BMSCs were co-cultured with SiHa and HeLa cells. Following miR-129 overexpression or knockdown, cell proliferation (CCK-8, colony formation), invasion (Transwell) and expression of E-cadherin, N-cadherin and MRP1 (Western blot) were examined. miR-129 expression was significantly higher in cervical cancer tissues than in normal controls (3.24&#xb1;0.56 vs. 1.00&#xb1;0.32, P<0.05). High miR-129 expression correlated with tumor size &#x2265;3 cm (P=0.009), FIGO stage III-IV (P=0.0034) and lymph node metastasis (P=0.038). High expression was associated with shorter OS (median: 28.4 vs. 46.2 months; HR=2.944, 95%CI:1.484-5.842, P=0.002). The AUC was 0.788 (95%CI:0.693-0.884), with a cut-off of 1.85 (sensitivity:76.9%, specificity:70.4%). BMSC co-culture upregulated miR-129 expression (SiHa: 3.52&#xb1;0.42 vs. 1.00&#xb1;0.08, P<0.01). miR-129 overexpression promoted proliferation, colony formation and invasion, while knockdown suppressed these phenotypes. Overexpression of miR-129 reduced E-cadherin (0.42&#xb1;0.05 vs. 1.00&#xb1;0.08, P<0.01) and increased N-cadherin (2.85&#xb1;0.22 vs. 1.00&#xb1;0.10, P<0.01) and MRP1 (2.46&#xb1;0.18 vs. 1.00&#xb1;0.09, P<0.01). Knockdown of miR-129 partially reversed BMSC-induced changes in these proteins. miR-129 is significantly upregulated in HPV-associated cervical cancer and shows potential as a diagnostic and prognostic biomarker. BMSCs upregulate miR-129 in cervical cancer cells and miR-129 modulates proliferation, invasion, EMT marker expression and MRP1 expression. These findings suggest that the BMSCs/miR-129 axis may contribute to cervical cancer progression, but further in-vivo and mechanistic studies are needed.

This paper critically examines livelihood recovery in the peripheral resettlement sites of Chennai, India, following the 2004 Indian Ocean tsunami. Despite the implementation of various gender-inclusive livelihood programmes as a part of disaster recovery, our longitudinal empirical research reveals a persistent gap between formal recovery measures and the everyday conditions required for livelihoods to become gender-inclusive and sustainable over time. Drawing on in-depth fieldwork, we explore how women respond in the long term to these limitations: not simply through individual coping strategies but also by developing and maintaining collective, care-centred practices, including adapting domestic spaces to work, sharing childcare responsibilities, and building networks of mutual support. These practices are often invisible in official recovery frameworks, yet they serve as critical choices for women both to sustain their livelihoods and perform their care roles. We build on Joan C. Tronto and Berenice Fisher's (1990) theory of caring to conceptualise these acts as forms of 'caring with', where care becomes a collective, political, and relational response to systemic exclusion. In doing so, we reposition disaster recovery not merely as a return to normalcy or economic productivity but also as a deeply social and gendered process that is shaped by care and solidarity.