Online medical consultation (OMC) services have gained considerable attention as integral components of telemedicine. Recently, artificial intelligence (AI) has been increasingly integrated into OMC platforms, facilitating enhanced consultation efficiency and clinical decision-making capabilities. Despite the availability and potential benefits of AI-driven OMC services, the public acceptance and willingness to pay (WTP) for AI-driven OMC services remain low. This study aimed to explore public perception of the AI-driven OMC services and to further evaluate the WTP for the service. We conducted semi-structured qualitative interviews with patients, caregivers, and healthcare professionals to explore factors influencing public acceptance and WTP for AI-driven OMC services. The study was informed by the theories of perceived risk and perceived benefit, which guided the development of the interview guide. All interviews were audio-recorded and transcribed verbatim. Data were analyzed using NVivo 15 with deductive thematic analysis guided by these theories, and coding was independently conducted and cross-checked by two researchers to ensure credibility and consistency. Thematic analysis of 20 in-depth interviews identified 2 main themes and 11 subthemes. Perceived risks and perceived benefits emerged as two key perspectives influencing participants’ acceptance and WTP. Psychological, privacy, social, functional, health, and financial risks reduced acceptance, whereas convenience, diversity, reliability, efficiency, and educational benefits promoted it. Reported WTP ranged from $0 to $30, with service-experienced participants generally reporting higher values than service-naive participants. This study identified the facilitators and barriers influencing public acceptance and WTP for AI-driven OMC services with theoretical constructs. Our findings offer valuable insights for the development and refinement of AI-driven OMC services, enabling more targeted pricing strategies and tailored services that can address the preferences and concerns of the public.

Background/Objectives: The therapeutic window for growth hormone (GH) therapy in late-pubertal males with advanced bone age (BA ≥ 15 years) is generally considered closed. Yet, open knee physes may indicate residual growth potential. This single-arm retrospective study evaluated the efficacy and safety of letrozole combined with GH in Chinese males with short stature, advanced BA (15 -< 18 years), and open knee physes. Methods: We included 139 male adolescents stratified by baseline BA: 15 ≤ BA < 16 (n = 24), 16 ≤ BA < 17 (n = 31), and 17 ≤ BA < 18 (n = 84). All received subcutaneous GH (0.05-0.07 mg/kg/day) and oral letrozole (2.5 mg/day). The primary outcome was the change in final adult height (FAH) from baseline predicted adult height (PAH). Longitudinal changes in height velocity and height SDS-BA were also assessed. Results: Among 75 patients who reached FAH, combination therapy was associated with a mean gain of 6.58 ± 3.46 cm over baseline PAH (p < 0.001). The gain was inversely correlated with baseline BA; the largest gain occurred in the 15 ≤ BA < 16 subgroup (10.04 ± 4.66 cm), significantly exceeding gains in the 16 ≤ BA < 17 (6.30 ± 3.01 cm) and 17 ≤ BA < 18 (5.70 ± 2.52 cm) subgroups (p < 0.001). In the youngest subgroup, FAH exceeded mid-parental height. The safety profile was acceptable; acne was the most common adverse event (46.76%), and no severe metabolic or endocrine disorders were observed. Conclusions: Letrozole plus GH therapy may be associated with improved FAH in late-pubertal males with advanced BA and open knee physes, with gains varying by baseline skeletal maturity and being numerically greatest in those with BA 15 -< 16 years. Knee epiphyseal status may be a useful consideration when individualizing therapy in this population.

Precision medicine is increasingly constrained not by a lack of molecular data but by the absence of frameworks that can translate multidimensional biological information into actionable clinical decisions. Multi-omics-driven precision medicine (MODPM) addresses this lack by integrating genomics, epigenomics, transcriptomics, proteomics, metabolomics, microbiome, and clinical context into a multiscale framework that links molecular mechanisms, tissue organization, and patient trajectories. In this review, we propose a conceptual framework for MODPM and examine how advances in multi-omics technologies, artificial intelligence (AI), and foundation models are reshaping disease modeling, drug development, and precision intervention. We summarize the biological contributions of major omics layers and discuss how AI supports cross-modal representation learning, contextual modeling, and perturbation-aware prediction. We highlight drug development as a key translational application of MODPM and further discuss its clinical relevance across three major disease contexts: cancer, autoimmune diseases, and metabolic disorders, including cardiometabolic and renal-metabolic diseases. These examples illustrate how MODPM can support target discovery, disease endotyping, treatment response prediction, and clinical monitoring by analyzing shared mechanisms such as immune dysregulation, metabolic remodeling, chronic inflammation, tissue microenvironmental changes, and gene-environment interactions. Across these settings, MODPM enables finer molecular stratification, the identification of pathway-dominant disease states, improved response prediction, and dynamic treatment monitoring. We also discuss key barriers to implementation, including data heterogeneity, limited cohort diversity, polygenic complexity, workflow constraints, cost, and ethical issues related to privacy, consent, and data ownership. Overall, the value of MODPM lies not in stacking additional data layers but in building a multiscale, continuously learnable framework to link biological heterogeneity to clinically interpretable and actionable decisions.

BACKGROUND:We identified significant gaps in perioperative pain management for sports medicine surgery at our hospital, including a high incidence of severe postoperative pain possibly due to a lack of preemptive analgesia. OBJECTIVE:We aimed to improve pain management outcomes by optimizing multimodal analgesia, which was implemented by the "Pain-Free Hospital" project at Peking University Third Hospital. STUDY DESIGN:A single-center retrospective observational study. SETTING:Peking University Third Hospital. METHODS:A total of 6,610 sports medicine surgery patients seen from July 2022 through August 2023 were divided into baseline (the first 3 months) and intervention (the subsequent 11 months) phases. Data on postoperative Numeric Rating Scale (NRS-11) pain scores and analgesic utilization were extracted from the hospital information system. Analgesic consumption was measured by the Analgesic Use Rate (AUR) and Number of Defined Daily Doses (DDDs). The DDD value of each drug is provided by the World Health Organization (WHO). The proportion of Days with Severe Pain (DSP, NRS-11 ≥ 7) was assessed on the first postoperative day and over the first 3 days. RESULTS:We addressed the gap in preemptive analgesia, with its implementation rate increasing from 0% to 53.8%. Etoricoxib (not approved for use in the United States by the FDA) use increased significantly (AUR: 13.4% to 53.2%; DDDs: 116.0 to 433.1), while pethidine use decreased significantly (AUR: 48.5% to 19.6%; DDDs: 47.1 to 17.0). These changes were accompanied by significant reductions in severe postoperative pain: DSP declined from 21.3% to 8.8% on the first postoperative day and from 10.8% to 4.8% over the first 3 days. LIMITATION:As a single-center retrospective study in a tertiary hospital in the People's Republic of China's capital, the regional patient pool may limit the generalizability of our findings. Additionally, the study did not account for the potential effect of COVID-19 on patient severity, surgical patterns, and analgesic prescription diversity during the pandemic. CONCLUSION:Our study demonstrates that implementing a structured perioperative protocol emphasizing preemptive analgesia and multimodal analgesia resulted in pain management effects, including decreased incidences of severe postoperative pain and reduced analgesic opioid consumption.

Exogenous progesterone supplementation is widely applied for the treatment of menopausal syndrome and assisted reproductive technology (ART). Nevertheless, conventional oral and injectable progesterone formulations suffer from low bioavailability, severe hepatic first-pass metabolism and injection-related pain. A novel progesterone-loaded ultrafine fiber patch was fabricated via solvent-free multi-nozzle melt electrospinning. Key fabrication parameters were systematically optimized to obtain uniform fibrous morphology and high drug loading capacity. In vitro drug release, ex vivo transdermal permeation, in vivo pharmacokinetics, skin adhesion performance and skin irritation tests were sequentially conducted for comprehensive evaluation. The average fiber diameter was 13.44 ± 3.46 μm with a low coefficient of variation. The patch realized sustained drug release for more than 168 hours after removal, together with prolonged (55.34 ± 36.95 h) elimination half-life. For patches loaded with 800 mg progesterone, the main pharmacokinetic parameters were determined as follows, Cmax = 101.44 ± 61.60 ng/mL, Tmax = 100.80 ± 35.27 h and AUClast = 4759.79 ± 1580.35 h·ng/mL, which showed promising potential for subsequent preclinical development. The patch maintained stable adhesion for 7 days with a 180° peel strength of 1.2 N/cm, and induced no obvious skin irritation. In vivo tissue distribution results revealed high progesterone accumulation in the skin (607.57 ± 39.77ng/g) for prolonged drug release. Moreover, the progesterone concentration in the uterus reached 23.17 ng/g, equivalent to the peak level during the luteal phase. This melt-electrospun fibrous patch is a promising transdermal delivery platform that integrates superior skin permeation, long-term sustained release, favorable adhesion property and excellent biosafety, showing a great potential for clinical translation.

Abstract Precision medicine is increasingly constrained not by a lack of molecular data but by the absence of frameworks that can translate multidimensional biological information into actionable clinical decisions. Multi‐omics‐driven precision medicine (MODPM) addresses this lack by integrating genomics, epigenomics, transcriptomics, proteomics, metabolomics, microbiome, and clinical context into a multiscale framework that links molecular mechanisms, tissue organization, and patient trajectories. In this review, we propose a conceptual framework for MODPM and examine how advances in multi‐omics technologies, artificial intelligence (AI), and foundation models are reshaping disease modeling, drug development, and precision intervention. We summarize the biological contributions of major omics layers and discuss how AI supports cross‐modal representation learning, contextual modeling, and perturbation‐aware prediction. We highlight drug development as a key translational application of MODPM and further discuss its clinical relevance across three major disease contexts: cancer, autoimmune diseases, and metabolic disorders, including cardiometabolic and renal–metabolic diseases. These examples illustrate how MODPM can support target discovery, disease endotyping, treatment response prediction, and clinical monitoring by analyzing shared mechanisms such as immune dysregulation, metabolic remodeling, chronic inflammation, tissue microenvironmental changes, and gene–environment interactions. Across these settings, MODPM enables finer molecular stratification, the identification of pathway‐dominant disease states, improved response prediction, and dynamic treatment monitoring. We also discuss key barriers to implementation, including data heterogeneity, limited cohort diversity, polygenic complexity, workflow constraints, cost, and ethical issues related to privacy, consent, and data ownership. Overall, the value of MODPM lies not in stacking additional data layers but in building a multiscale, continuously learnable framework to link biological heterogeneity to clinically interpretable and actionable decisions.

OBJECTIVES:Surgical site infections (SSIs) represent a significant postoperative challenge. Although timely perioperative prophylaxis with cephalosporins is essential to prevention, adherence to the recommended 30-60 min administration window may be challenging with traditional intravenous infusion (IVI) in settings with high surgical turnover, as is the case in China. Intravenous push (IVP) has been proposed as a more time-efficient alternative. This scoping review aims to map the available evidence comparing IVP with IVI for perioperative cephalosporin administration across four domains: safety, pharmacokinetics/pharmacodynamics (PK/PD), efficacy, and economic impact. METHODS:A systematic search was conducted across PubMed, Embase, Web of Science, the Cochrane Library, and gray literature up to February 2026. Data were systematically charted and extracted using a standardized form. RESULTS:Of the 14 included sources, only 3 were peer-reviewed comparative studies; the remaining 11 (78.6%) were gray literature documents. Among the gray literature, 72.7% (8/11) permitted or recommended IVP for cephalosporin prophylaxis; however, this proportion reflected practice patterns of heterogeneous methodological rigor. The 3 peer-reviewed studies focused on the safety, PK/PD, and economic outcomes. Two studies-in orthopedic and bariatric surgery, respectively-found no significant difference in adverse event rates between IVP and IVI, though both were limited by small samples. A single small study suggested similar PK/PD target attainment between IVI and IVP cefazolin. No study directly compared SSI rates between the two routes. One study suggested potential cost savings with IVP, but the evidence was dated and based on limited patient numbers. CONCLUSIONS:The available evidence for IVP is predominantly derived from gray literature, while peer-reviewed articles suggest that safety and PK/PD profiles do not differ markedly from IVI in the limited populations, surgical procedures, and agents studied; economic data are suggestive but dated. Direct comparative data on clinical efficacy outcomes, such as SSI rates, are absent. Well-powered, multi-center comparative studies comparing IVP and IVI with SSI as a primary endpoint are needed.

Oral anticoagulant therapy is widely used for the prevention and treatment of thromboembolic diseases, yet it imposes significant burdens on patients’ health-related quality of life (HRQoL). This study aimed to compare the performance of two generic preference-based measures (GPBMs), the EQ-5D-5 L and SF-6D, in assessing HRQoL among patients receiving oral anticoagulant therapy, and to further explore the factors influencing their HRQoL. A multicenter, cross-sectional survey was conducted across 14 hospitals in China. Patients receiving oral anticoagulant therapy were enrolled and completed the EQ-5D-5 L, SF-6D, anticoagulation satisfaction (assessed by DASS), health literacy (assessed by HLS-SF9), and self-efficacy (assessed by NGSES-SF). Ceiling and floor effects, the association and agreement between the two measures, known-group validity and multi-factor analysis were assessed. A total of 379 patients receiving oral anticoagulant therapy were included in the analysis. The mean (standard deviation) utility scores were 0.837 (0.187) for the EQ-5D-5 L and 0.729 (0.132) for SF-6D. The EQ-5D-5 L demonstrated a higher ceiling effect (21.11

Accurate quantification of circulating androgens, particularly the biologically active free fraction, is analytically challenging due to ultra-low concentrations and extensive protein binding in human serum. Here, we developed and validated an integrated ultrafiltration-LC-MS/MS workflow for simultaneous quantification of nine endogenous serum free androgens. Free fractions were isolated by phosphate-buffered saline dilution followed by temperature-controlled centrifugal ultrafiltration (37 °C) using pretreated regenerated-cellulose membranes to minimize non-specific adsorption. Ultrafiltrates were purified and enriched by magnetic solid-phase extraction (MSPE), and in-situ oximation derivatization with a quaternary aminooxy reagent was applied to enhance electrospray response. The validation data demonstrated pg/mL-level sensitivity with LLOQ of 0.5-10 pg/mL, good linearity (r > 0.99), acceptable matrix effects, and high recovery. The method was applied to serum samples from 86 female patients (21-50 years) undergoing infertility evaluation, showing higher free testosterone, 11β-hydroxyandrostenedione, and androstenedione in an AMH-enriched PCOS-suspicion subgroup versus a control group. Overall, this ultrafiltration-MSPE-in-situ derivatization LC-MS/MS method provides a practical and robust platform for multiplexed measurement of serum free androgens at pg/mL levels for clinical research applications.

Acute kidney injury (AKI) is a common syndrome among critically ill patients with high incidence and high mortality, characterized by elevated serum creatinine (SCr) and uric acid (UA) levels. Given that xanthine is a key intermediate in nucleotide metabolism and the direct precursor of UA, profiling of nucleotide metabolic dysregulation in AKI remains largely unexplored. Using ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS), we simultaneously quantified 29 nucleotide intermediates both in plasma and matched urine samples from 58 propensity score-matched pairs of AKI and non-AKI (NAKI) critically ill patients. Compared with NAKI controls, AKI patients showed decreased levels of 7 urinary nucleotide intermediates and increased levels of 4 plasma metabolites. Urinary nucleotide levels correlated more strongly with SCr and estimated Glomerular Filtration Rate (eGFR) than their plasma counterparts. Elevated urinary xanthine and other 5 metabolites were identified as protective factors for AKI, while elevated plasma adenine, thymine and cytosine were risk factors. A combination of urinary guanine and xanthine with plasma thymine discriminated AKI risk with an area under the curve (AUC = 0.880, 95% CI = 0.800-0.934). Interestingly, alterations in nucleotide intermediates influenced AKI occurrence mediated by SCr, eGFR, UA, urea and aspartate aminotransferase (AST), with hypoxanthine and thymidine exerting specifically through AST. In summary, dysregulated nucleotide metabolism was closely associated with AKI onset and participated in kidney-liver crosstalk through modulation of liver function, providing new mechanistic insights into AKI pathogenesis.

OBJECTIVE:To develop an evidence-based patient version of guideline (PVG) for fibromyalgia, aiming to improve patients' understanding of disease symptoms and therapeutic options and to enhance their self-management abilities. METHODS:Following the World Health Organization Handbook for Guideline Development (2014), a multidisciplinary working group was established, including patient representatives, physicians, pharmacists, nurses, and methodologists. The process comprised (a) systematic retrieval of clinical practice guidelines and expert consensus statements to establish the evidence base; (b) integration of large language models (LLMs) with expert review to identify and refine key patient-centered concerns; (c) a three-round Delphi consensus guided by the Grading of Recommendations Assessment, Development and Evaluation approach to finalize clinical questions and formulate recommendations; and (d) evaluation of understandability using the Patient Education Materials Assessment Tool for Print Materials (PEMAT-P). RESULTS:The final PVG covers 13 clinical questions across seven domains, including disease awareness, diagnostic evaluation, pharmacological and non-pharmacological interventions, and long-term management. All recommendations were rated as strong. The guideline emphasizes the importance of pharmacological management, emotional regulation, and exercise in the comprehensive management of fibromyalgia. The PEMAT-P assessment showed an understandability score of 100%. CONCLUSIONS:Developed collaboratively by a multidisciplinary team and patient representatives, this PVG is based on 13 evidence-based fibromyalgia guidelines. Combining LLMs with expert review enhanced question generation and readability. The PVG provides a practical and accessible tool to support early self-management in fibromyalgia.

BACKGROUND:With the extensive utilization of immune checkpoint inhibitors (ICIs) across various cancers, ICIs-related thyroid dysfunction (ICI-TD) has become a growing concern in clinical practice. This study aimed to devise an individualized management strategy for ICI-TD to enhance the early identification and proactive management in cancer patients. METHODS:We designed and conducted a three-phase study. Initially, we analyzed the influencing factors through a systematic review and meta-analysis, which adhered to the Preferred Reporting Items for Systematic Reviews and Meta-analyses (PRISMA) guidelines. Moreover, the study protocol was registered with PROSPERO (CRD42019131133). Subsequently, prediction models for ICI-TD were developed utilizing 11 algorithms based on the real-world cohort data from July 20, 2018 (the approval date of the first ICIs, Pembrolizumab in China), to October 31, 2022. Considering discrimination, calibration, and clinical utility, we selected the model with the best performance for web calculator development. Finally, individualized management strategies for ICI-TD were proposed by combining evidence-based analysis with practical considerations. RESULTS:The systematic review encompassed 21 observational studies involving 4,145 patients, revealing associations between ICI-TD and factors such as female gender, age, receipt of Pembrolizumab (versus other ICIs), and baseline levels of thyroid-stimulating hormone, free thyroxine, and antithyroid antibodies. In the prediction model development phase, 621 participants were enrolled, with 36 patients developing ICI-TD. The model based on the LightGBM algorithm demonstrated superior performance, leading to the development of a web calculator. Based on these findings and existing guidelines, individualized monitoring and treatment pathways for pharmacists were devised. CONCLUSION:This study offers comprehensive insights into managing ICI-TD, potentially enhancing tailored cancer immunotherapy management.

Background:In therapeutic drug monitoring (TDM) of tacrolimus, trough concentration is the most commonly used monitoring indicator, which has limited association with clinical outcomes. Intrapatient variability (IPV), as a novel monitoring marker for describing the extent of fluctuation in blood concentration, holds promise as an additional monitoring tool. The effect of tacrolimus IPV on Chinese kidney transplantation outcomes is unclear, and the aim of this study is to explore the association between IPV and poorer outcomes of Chinese kidney transplantation recipients and to discuss the influence of genotype. Methods:A total of 152 patients were enrolled, whose kidney transplantation operations were carried out from January 2015 to February 2022 in Peking University Third Hospital. IPV was calculated by coefficient of variation (CV) of tacrolimus whole blood concentrations during 6 and 12 months after transplantation. Clinical outcomes were analyzed between patients with high IPV and low IPV, including graft loss, acute rejection (AR), elevated serum creatinine (Scr), infection, hemogram abnormality and electrolyte disturbance. Moreover, the associations were compared according to different genotypes. Results:High IPV was associated with AR [hazard ratio (HR) =3.420; 95% confidence interval (CI): 1.142-10.245], hyperuricemia, and hypocalcemia (P<0.05). In cytochrome P450 (CYP)3A5 nonexpressers, high IPV was associated with graft loss, elevated Scr, hyperuricemia, hypocalcemia, hypomagnesemia and leukopenia (P<0.05), whereas no significant association was detected in CYP3A5 expressers (P>0.05). Conclusions:IPV is associated with clinical outcomes such as AR in Chinese kidney transplantation recipients. More attention should be paid to the association between high IPV and adverse outcomes in CYP3A5 nonexpresser recipients.

Introduction: Bruton's tyrosine kinase inhibitors (BTKis) have transformed the treatment landscape of B-cell malignancies, demonstrating both proven efficacy and an acceptable safety profile. BTKis are primarily metabolized by cytochrome P450 (CYP) 3A4, making it susceptible to drug-drug interactions (DDIs) with CYP3A4 inhibitors or inducers like voriconazole.Therapeutic drug monitoring (TDM) quantifies drug concentrations in patient samples using modern analytical techniques and computational methods, enabling individualized dosing that has significantly improved patient outcomes. This study aimed to describe the effect of voriconazole on BTKis concentrations and guide dose adjustment based on TDM when used in combination. Methods: Four patients with B-cell malignancies received BTKis in combination with voriconazole [200 mg twice a day (BID)] for the treatment of fungal or respiratory infections. The regimens included ibrutinib [560 mg once a day (QD), reduced to 140 mg QD], zanubrutinib (160 mg BID, reduced to 80 mg QD), orelabrutinib (150 mg QD, reduced to 50 mg QD), and acalabrutinib (100 mg BID, reduced to 100 mg QD). Dose reductions for ibrutinib and zanubrutinib followed prescribing information, while adjustments for orelabrutinib and acalabrutinib-lacking formal recommendations-were based on clinical judgment. TDM of BTKis was carried out by liquid chromatography tandem mass spectrometry (LC-MS/MS). Results: TDM revealed that trough concentrations of BTKis remained within a comparable range before and after dose adjustments. Trough concentrations of BTKis before and after dose reduction were as follows: ibrutinib, 4.28 mg/L and 5.27 mg/L; zanubrutinib, 15.85 mg/L and 18.96 mg/L; orelabrutinib, 21.91 mg/L and 25.05 mg/L; and acalabrutinib, 12.89 mg/L and 13.61 mg/L. These findings support the appropriateness of dose reduction in the presence of CYP3A4 inhibition. And no adverse drug events (ADEs) were observed during the treatment with BTKis in combination with voriconazole. Conclusion: The study suggests that coadministration of BTKis with CYP3A inhibitors can significantly affect its pharmacokinetic properties, and TDM has been shown to optimize the individualized dose adjustments of BTKis in the presence of DDIs. We are currently conducting a prospective real-world study to further elucidate exposure-response relationships, aiming to transition BTKi therapy from empirical to precision medicine. These efforts will provide valuable insights for optimizing dosing regimens, guiding personalized treatment decisions, and improving clinical outcomes.

Objectives: This bibliometric analysis investigates recent research trends in biologics and small molecules for treating inflammatory bowel disease (IBD) based on literature from the past decade. Methods: This cross-sectional study involved analyzing data retrieved from the Web of Science Core Collection (WoSCC) database to examine the evolution and thematic trends of biological agents and small-molecular drugs for IBD conducted between 1 January 2014, and 20 September 2024. VOSviewer software was utilized to assess co-authorship, co-occurrence, co-citation, and network visualization, followed by a further discussion on significant sub-themes. Results: From 2014 to 20 September 2024, the annual number of global publications increased by 23%, reflecting an acceleration in research activity. The journal “Inflammatory Bowel Diseases” published the highest number of manuscripts (579 publications) and garnered the most citations (13,632 citations), followed by the “Journal of Crohn’s & Colitis” (480 publications) and “Alimentary Pharmacology & Therapeutics” (250 publications). The United States led in productivity with 1943 publications and 66,320 citations, with UC San Diego (291) and authors Sandborn and Vermeire (180) topping the list. The co-occurrence cluster analysis of the top 100 keywords resulted in the formation of six distinct clusters: Disease Mechanisms, Drug Development, Surgical Interventions, Therapeutic Drug Monitoring (TDM), Immunological Targets, and Emerging Therapies. Burst terms (TNF-α inhibitors, JAK inhibitors, and trough-level optimization) highlight trends toward personalized biologics and small-molecule regimens. Conclusions: The bibliometric analysis indicates that IBD therapeutic research and clinical applications focus on biologics and small molecules, with research trends leaning toward precise therapy conversion or the combination in non-responders. Future work will assess monotherapy, the combination, and conversion therapies and investigate new drugs targeting inflammatory pathways.

The accumulation of classic or 11-oxygenated androgens is a hallmark of polycystic ovary syndrome (PCOS). Traditional electrochemiluminescence immunoassays (ECLIA) are limited in their ability to measure a broad spectrum of androgens and may suffer from potential cross-reactivity, leading to inaccurate results. In this study, we developed a highly sensitive HPLC-MS/MS method for the simultaneous determination of nine androgens, including four classic and five 11-oxygenated androgens in human serum. Magnetic solid-phase extraction (MSPE) using magnetic graphene oxide nanoparticles followed by in-situ derivatization further improved the detection efficiency. A novel quaternary ammonium aminooxy (QAO) reagent was employed as the derivatization agent, featuring a permanently charged MS-tag and an aminooxy group that reacts with steroid ketone groups. This derivatization significantly enhanced MS sensitivity compared to traditional derivatization agents. The linear ranges for different targets were set based on clinical use and the lower limit of quantification (LLOQ) of the target androgens ranged from 0.005 to 1 ng/mL. The calibration curves exhibited excellent linear correlation (r ≥ 0.99) across the validated ranges for all the analytes. The intra- and inter-run precision was below 9.8 % and 9.7 % for the lower limit of quantification (LLOQ), and 7.5 % and 6.9 % for quality controls (QCs). Intra- and inter-day relative errors for LLOQ ranged from -8.2 % to 10.7 %, and from -7.4 % to 13.8 % for QCs. Extraction recoveries varied from 41.8 % to 80.2 %. The method was applied to 107 clinical samples, demonstrating it as a fast, sensitive and accurate method for androgens’ quantification. By offering the ability to detect and quantify a broader range of androgens, including those not measurable by ECLIA, and avoiding potential cross-reactions. This method provides a superior analytical tool for the diagnosis and management of PCOS in clinical settings.

Chemical isotope labeling (CIL) coupled with liquid chromatography-mass spectrometry (LC-MS) is a highly effective approach for comprehensive metabolomic profiling, offering excellent quantification precision. An isotope reagent, d(4)-dimethylethylenediamine, can be readily synthesized, and together with its d(0)-labeled counterpart, offered a straightforward and reliable method for labeling bile acids with carboxyl functional groups. The light and heavy isotope-tagged bile acid analogs co-eluted and were detected concurrently via mass spectrometry, allowing for precise quantification and robust metabolite identification. Relative quantification demonstrated an average relative standard deviation (RSD) of peak area ratios of 15.4 % over 3 days, with more than 89.0 % of the bile acids exhibiting peak area ratio values between 0.8 and 1.2. In total, 108 bile acids were confidently or tentatively identified in pooled human serum samples, with 55 of them verified using authentic reference standards. The method was further employed to explore bile acid metabolic alterations in human serum associated with intrahepatic cholestasis of pregnancy (ICP) based on a case-control study. Fifteen bile acid species demonstrated statistically significant alterations (p < 0.001) when comparing the ICP group to the control group, with each displaying a variable importance in projection (VIP) score exceeding 1.5 in partial least squares discriminant analysis (PLS-DA). Ten metabolite ratios were screened as the potential biomarkers for ICP diagnosis, achieving sensitivities greater than 92.0 % and specificities greater than 94.0 %. The presented strategy marbly enhanced analytical coverage and robustness, offering significant utility in discovering reliable potential biomarkers for various diseases associated with bile acid metabolism disorders.

Patients with sepsis are at a heightened risk of long-term cognitive impairment, including neurodegenerative diseases; however, the underlying pathophysiological mechanisms remain incompletely understood. This study examines key genes associated with sepsis and Alzheimer’s disease (AD), as well as their potential molecular mechanisms. We downloaded the GSE135838 dataset from the Gene Expression Omnibus (GEO) database and performed comparative analysis of differentially expressed genes (DEGs) using the AlzData database to identify co-expressed DEGs. Functional and protein–protein interaction (PPI) network analyses were used to identify hub genes and their associated molecular mechanisms. Animal experiments were conducted to validate the role of the central gene C5aR1 in the pathological processes of sepsis-related cognitive impairment, blood–brain barrier (BBB) disruption, and microglial activation. Co-culture experiments were performed to assess the protective effect of C5aR1 against inflammation-induced neuronal damage. In GSE135838, 25 DEGs exhibited consistent expression changes in the brain tissue of AD patients. Notably, LYZ, C5AR1, ZFP36, MPZL2, APOL4, CD163, SERPINA3, and CCL2 showed significant differential expression in the cortex and hippocampus of AD patients. KEGG pathway enrichment analysis revealed that among the 14 pathways meeting the criteria, the TNF signaling pathway demonstrated the highest significance. Key intersections of multiple GO enrichment terms included IL-6, ICAM1, CLEC4E, and PCK1. The top ten hub genes identified from the PPI network analysis included IL6, CCL2, ICAM1, CXCL1, CD163, C5AR1, SOCS3, CLEC4E, HSPB1, and HSPA1A. Pharmacological inhibition of the hub gene product C5aR1 using PMX205 improved cognitive and emotional dysfunction in CLP-induced septic mice and reduced BBB damage and microglial activation. Inhibition of C5aR1 also alleviated microglia-induced neuronal injury. In summary, the neuroimmune dysregulation caused by sepsis is correlated with potential pathological mechanisms in AD. This study provides additional molecular evidence for potential biomarkers and therapeutic targets for drug intervention in the risk of AD among sepsis survivors.