Large language model (LLM)-based conversational agents are increasingly used in healthcare, yet their capacity to support genuine multi-turn dialogue remains underexplored. In oncology, where patients and caregivers experience complex informational and emotional needs throughout the disease trajectory, conversational agents may support information provision, symptom consultation, and emotional assistance. However, research specifically examining multi-turn conversational agents designed for cancer patients and informal caregivers remains limited. This scoping review aimed to map the research landscape of LLM-based multi-turn conversational chatbots developed for cancer patients and informal caregivers, focusing on system design, intervention purposes, evaluation approaches, safety considerations, and transparency of LLM-related components. This scoping review followed the Joanna Briggs Institute methodology and PRISMA-ScR guidelines. Eight databases were searched: PubMed, Embase, Scopus, Web of Science, CINAHL, and PsycINFO were searched for studies published between January 2022 and January 2026, with supplementary searches conducted in IEEE Xplore Digital Library and ACM Digital Library in May 2026. Studies were included if they described LLM-based chatbots designed for cancer patients or informal caregivers that supported multi-turn conversational interaction. Two reviewers independently conducted the study selection and data extraction. Eight studies met the inclusion criteria. Most studies focused on prototype development, with limited research evaluating clinical outcomes. ChatGPT-based models were the most used LLMs, and retrieval-augmented generation techniques were applied in several studies. Chatbots were primarily designed for emotional support or information provision. Evaluation approaches varied widely, including response quality, psychological outcomes, and user experience. However, no studies evaluated interaction-level characteristics such as conversational continuity or context retention, and only two studies reported any conversational memory mechanism. Reporting on safety risks, mitigation strategies, prompt design, model parameters, and adherence to LLM reporting guidelines was often limited or absent. This scoping review identified only 8 studies on LLM-based multi-turn conversational chatbots for cancer patients and informal caregivers. The field remains at an early stage characterized by prototype-oriented development, heterogeneous design and evaluation approaches, and inconsistent safety and transparency reporting. Future development should prioritize genuine conversational capability, safety management and transparent reporting.
This study investigates the size and shape variation of cochleae in the Russian population. Also, introduces a new cochlear parameter that defines the longest straight segment of the basal turn, with the implication of a safe cochlear implant electrode insertion. A medical-grade, CE-marked DICOM viewer (OTOPLAN) was used to measure cochlear parameters, including A- and B-values, which define the length and width of the basal turn. The shape of the cochlear basal turn was estimated by the ratio between B-and A-values. The new cochlear parameter, I-value, defines the effective straight segment of the basal turn as the longest straight line connecting the cochlear entrance to the lateral wall at the opposite end, passing tangentially to the inner wall. Ratio between B-, and A-value determines the shape of the cochlear basal turn, with the cut-off value ≥0.75 for round-shaped and <0.75 for elliptical-shaped cochlear basal turn. CT scans of the temporal bone from 112 ears with normal inner ear anatomy were used in this study. The cochlear size of the Russian population, as measured by A-value, ranged from 8.04-10.03 mm, with a mean of 9.03 mm. The B-value ranged from 5.84-7.59 mm, with a mean of 6.70 mm. A round-shaped cochlear basal turn was observed in 48% of the population, leaving the remaining 52% of the population with an elliptical-shaped basal turn. The I-value that defines the effective straight segment of the basal turn ranged from 6.7-9.6 mm, with an average of 8.3 mm. The trend line indicates that the I-value is approximately 0.7-0.8 mm shorter than the A-value. For the first time in the literature, cochlear size, as measured by A-value, is presented for the Russian-speaking population. The round-shaped cochlear basal turn is seen in 48% of the population. The new cochlea parameter, the I-value, is approximately 0.7-0.8 mm shorter than the A-value.
Large language model (LLM)-based virtual patients are increasingly used to scale history-taking practice in undergraduate and postgraduate medical education. For clinical simulation, reliability requires not only avoidance of harmful content but also role-consistent case fidelity, dialogue continuity, and adherence to constraints on what the simulated patient should disclose. We evaluated these dimensions as an operational robustness benchmark, not as evidence of deployment readiness. We systematically stress-tested a Japanese LLM-based virtual patient under six robustness conditions: clean inputs, noise, direct contamination, direct contamination with defense, indirect contamination, and indirect contamination with defense. Case fidelity was measured using the slot-level F1 score, a 0-to-1 measure combining precision and recall for expected case-history elements, excluding the opening greeting turn. Information coverage was measured using turn-target hit rate, defined as the fraction of prespecified target slots elicited at the intended interview turns. Proxy constraint-adherence outcomes included refusal, clarification, forbidden information leakage, contradiction, and role drift. Under clean inputs, case fidelity was high (0.947; 95% confidence interval (CI), 0.940-0.955), and turn-target hit rate was 0.946 (95% CI, 0.932-0.959). Noise preserved overall case fidelity (0.941; 95% CI, 0.934-0.949) but reduced turn-target hit rate (0.823; 95% CI, 0.806-0.841). Direct contamination caused severe degradation in fidelity (0.098; 95% CI, 0.052-0.144) and turn-target hit rate (0.077; 95% CI, 0.034-0.119). With preprocessing defense, performance returned to near-clean levels (fidelity, 0.945; 95% CI, 0.938-0.952; hit rate, 0.942; 95% CI, 0.930-0.955). Indirect contamination showed near-clean fidelity (0.946; 95% CI, 0.938-0.954) and hit rate (0.952; 95% CI, 0.940-0.965), with minimal additional benefit from defense. Refusal and clarification rates were 0 across all conditions, and role drift events were not observed. However, forbidden information leakage occurred at approximately 0.60 events per episode under the clean condition (0.600; 95% CI, 0.502-0.698), indicating incomplete constraint adherence despite high case fidelity. In this controlled sequential virtual patient benchmark, direct prompt contamination was the dominant failure mode in terms of case fidelity and information coverage, while noise primarily reduced target-slot acquisition. A simple preprocessing defense mitigated direct-contamination effects on fidelity and information coverage, but persistent forbidden-information leakage indicates that additional safeguards and external validation are required before claims of safe clinical or educational deployment can be made.
The role of cover crops in promoting the abundance of beneficial soil microbes is increasingly gaining attention in tree crops. However, soil fungi remain comparatively understudied despite their roles in nutrient cycling and soil organic matter turnover, functions that are directly influenced by cover crops and may, in turn, affect the cash crop. This study compared the effects of cover crop mixtures to a weedy control on soil bacterial and fungal communities in young (< 10 years) and old (> 20 years) citrus orchards-with different lengths of cover cropping-across three soil depths over 2 years. Soil fungi responded to cover crops, whereas soil bacteria showed minimal changes in both orchards. In the young orchard, non-legume cover crop mixtures altered soil fungal communities three months after planting and after 2 years, both cover crop mixtures resulted in distinct community shifts relative to the control. In the old orchard, soil depth shaped fungal communities, with the strongest cover crop effects in the topsoil and subsurface soil. In both orchards, cover crops enriched potentially plant-beneficial microbial taxa, including arbuscular mycorrhizal fungi, highlighting the importance of cover crops as a management tool for driving fungal-mediated ecological processes and soil health benefits in orchard systems.
School exclusion in England disproportionately affects pupils with social, emotional, and mental health needs. Tabletop roleplaying games (TTRPGs) may strengthen social emotional learning but are rarely evaluated in UK state schools. Young Dragons, a Dungeons & Dragons based programme, was developed to support emotional regulation, teamwork, and engagement among pupils at risk of exclusion in two London boroughs. A convergent mixed methods realist evaluation was delivered across ten schools. Pupils aged 9-16 attended weekly one-hour sessions for 6-8 weeks in small, consistent groups. Pre/post pupil surveys captured wellbeing, self-concept, peer relations, school belonging, and loneliness; matched pair change (n = 22) used Wilcoxon signed rank tests. Semi-structured interviews with school staff and facilitators and observation of multiagency meetings explored implementation and perceived impact. Findings were integrated using joint displays and context-mechanism-outcome (CMO) mapping. No statistically significant within person change was detected across matched items (all p > 0.10). Distributions showed heterogeneous trajectories: many pupils reported better mood, anger regulation, and confidence, while a minority shifted toward greater disengagement or loneliness. Exploratory analysis of routine school records (n = 30) showed a significant reduction in mean suspensions from 0.7 pre-intervention to 0.0 post-intervention (p = 0.022) and a modest, non-significant increase in mean attendance from 90.6% to 92.7% (p = 0.069). Qualitative accounts described strong engagement, psychological safety, and visible gains in self-management, turn taking, and teamwork, with positive spillover into classroom behaviour when groups were stable and facilitation was consistent. Delivery challenges included timetable pressures, space constraints, and stigma around targeted provision. Integration identified skilled facilitation, small-group safety, and structured reflection as key mechanisms enabling co-regulation, perspective taking, and belonging. Young Dragons was feasible and acceptable in high need school settings. Benefits were mechanism-consistent for many participants but contingent on context and facilitator quality. These pilot data justify a larger, controlled evaluation to test causal pathways and longer-term outcomes for inclusion and emotional wellbeing.
The human RNA polymerase II-associated factor 1 complex (PAF1c) functions in transcriptional elongation and mRNA maturation. PAF1c is composed of several subunits: PAF1, CDC73, LEO1, CTR9, RTF1, and SKIC8. Besides its role in transcription, PAF1c subunits have been reported to be associated with tumorigenesis through maintaining cancer genome stability. In this study, we show that depletion of PAF1c leads to a pronounced accumulation of R-loops, which in turn results in an increase in DNA damage. Moreover, we confirmed that PAF1c deficiency increases the cytotoxicity of several DNA-damaging agents, including hydroxyurea (HU), cisplatin (CDDP), methyl methanesulfonate (MMS), and bleomycin (BLM). Unexpectedly, PAF1c depletion confers tolerance specifically to topoisomerase inhibitors, such as camptothecin (CPT), etoposide (ETOP), and doxorubicin (DOX). Further investigation revealed that the resistance to topoisomerase inhibitors induced by PAF1c depletion occurred specifically in G1-but not S-phase cells. Mechanistically, PAF1c depletion paradoxically decreases CPT-induced accumulation of R-loops by impeding mRNA elongation in G1-phase cells, thereby reducing CPT-induced cell death. Collectively, our findings demonstrate that loss of PAF1c subunits not only promotes genomic instability through R-loop accumulation but also alters cellular responses to DNA-damaging agents, conferring resistance particularly to topoisomerase inhibitors. This study underscores the critical role of PAF1c in maintaining genome stability and provides a rationale for developing new therapeutic strategies in cancer treatment.
Access to essential medicines is a fundamental human right and a critical pillar of effective healthcare. In Africa, armed conflicts severely weaken health systems, disrupting the availability of essential medicine, leading to gaps in patients' treatment and, in turn, posing a serious risk to overall public health. Therefore, this study aimed to identify the impact of armed conflicts on the availability of essential medicines as a cornerstone of healthcare across African regions, and quantify the extent of this disruption over time and by country. In this systematic review and meta-analysis, data were retrieved from published articles accessible in PubMed, Semantic Scholar, and grey literature covering the period from 1985 to 2025 and 2001 to 2024 for system impact and medicines availability studies, respectively. The literature search was conducted from January to May 2025. Following the Preferred Reporting Items for Systematic Reviews and Meta-analysis guidelines, studies were independently screened by two reviewers and included if they contained information on medicine availability or its relation to health systems in conflict zones and were published in English. The quality of studies was evaluated using the Joanna Briggs Institute criteria. Pooled estimates of medicine availability and their 95% CIs were obtained using a random-effects analysis. Heterogeneity was assessed using the I² statistic. The risk of bias and small study effects were assessed with funnel plots and Egger's test. Additionally, the leave-one-out sensitivity test and influence diagnostics test were considered to evaluate study-level impact. The review included data from 1,581 health facilities and 989 essential medicines across eight conflict-affected African countries. The pooled availability of essential medicines calculated from the data obtained from 41 studies conducted in African countries shouldering frequent armed conflict was estimated at 55% (95% CI, 47-63), with substantial heterogeneity across studies (I² = 93%, p < 0.001). Country-level analysis showed the highest availability in the Central African Republic (79%; 95% CI, 49-95) and the lowest in Nigeria (40%; 95% CI, 1-78). A temporal decline in medicine availability was observed, from 76% (2001-2010) to 46% post-2020. No significant publication bias was detected (funnel plot asymmetry p = 0.9560; Egger's test p = 0.494). Additionally, the influence diagnostic test and the leave-one-out sensitivity analysis indicated that observed heterogeneity likely reflects methodological differences and the sample size across the studies rather than bias or outliers. The systematic review revealed that armed conflicts have profoundly compromised health system functionality through the destruction of healthcare infrastructure, closure of medical facilities, displacement of the health workforce, pervasive insecurity and psychological distress among healthcare providers, and disruptions in supply chains and transportation networks, all of which have adversely affected the accessibility of essential medicines. On average, only 55% of essential medicines were available in conflict-affected regions of Africa, which was below the WHO benchmark of 80%. This finding underscores the severe threat that armed conflict poses to medicine availability and, consequently, to increased indirect morbidity and mortality. Additionally, the conflict undermines medicine availability through multiple, often interconnected mechanisms, indicating the need for a coordinated, multisectoral approach to ensure continuous access to essential medicines. Therefore, strengthening healthcare infrastructure in conflict settings is critical. Furthermore, the international community should enforce accountability mechanisms for violations affecting healthcare services, while donors and regional humanitarian assistance and emergency response mechanisms should increase investment and medicine supply in resilient medicine supply systems capable of maintaining availability during periods of armed conflict. The study protocol was registered in PROSPERO (CRD420251154811), the International Prospective Register of Systematic Reviews, maintained by the National Institute for Health and Care Research (NIHR).
Antiviral immunity profoundly impacts host metabolism, which can, in turn, modulate immune responses and influence disease pathology. The liver orchestrates systemic bile acid (BA) metabolism, a pathway disrupted in chronic liver diseases such as viral hepatitis. BAs are increasingly recognized for their immunomodulatory properties. Thus, improved understanding of the interplay between immunity and BA metabolism may reveal novel therapeutic avenues. Using lymphocytic choriomeningitis virus (LCMV) as a model, we investigated the interplay between chronic virus infection, BA metabolism, and immunity. Chronic LCMV infection increased BA levels and shifted circulating and liver BA composition toward host-derived, conjugated BAs. Hepatic BA transport and synthesis genes were broadly downregulated, in part depending on CD8+ T cells. Pharmacological inhibition of the main hepatic transporter of conjugated BAs, NTCP (Slc10a1), increased hepatic damage, while combined genetic disruption of the BA transporters Slco1a1, Slco1a4, and Slco1b2, responsible for the hepatic reuptake of unconjugated BA, reduced liver pathology and impaired antiviral CD8+ T cell responses. These findings reveal a reciprocal interplay between BA metabolism and CD8+ T cells, expanding our understanding of adaptive immunity in viral hepatitis. They also highlight how immunometabolic changes in liver disease may affect adaptive immune responses against infections.
Neodymium oxide (Nd2O3), a rare-earth oxide, has been shown to induce lung fibrosis. Our previous research indicated that glycolysis plays a key role in the occurrence and development of lung fibrosis following exposure to Nd2O3. Nevertheless, the role of lactate-mediated histone lactylation, a metabolic product of glycolysis, in lung fibrosis remains unclear. In the present study, we used a co-culture model of macrophage-like cells (THP-M) treated with Nd2O3 and human embryonic lung fibroblasts (HELF). We found that Nd2O3 significantly increases glycolytic activity, lactate production, and global protein lactylation (pan-Kla), particularly the lactylation of histone H4 at lysine 12 (H4K12la) in macrophages. Chromatin immunoprecipitation sequencing and chromatin immunoprecipitation-quantitative polymerase chain reaction showed that lactylation modification in the promoter region of the phosphoinositide-3-kinase regulatory subunit 2 (PIK3R2) gene in macrophages was significantly enriched. This enrichment was accompanied by upregulation of PIK3R2, activation of the PI3K/Akt/mTOR-signaling pathway, and increased secretion of transforming growth factor-β1 (TGF-β1). Further experiments showed that knockdown of PIK3R2 reversed the Nd2O3-induced profibrotic phenotype in macrophages, thereby significantly inhibiting TGF-β1 secretion and fibroblast activation and proliferation. Treatment with 2-deoxy-D-glucose (2-DG) inhibited glycolytic lactate production in THP-M cells, leading to reductions in overall histone lactylation, H4K12la levels, PIK3R2 expression, and TGF-β1 secretion. In addition, 2-DG also suppressed the PI3K/Akt/mTOR pathway and reversed fibrosis. Altogether, our findings reveal the role of histone lactylation in Nd2O3-induced macrophage-fibroblast interactions and elucidate the molecular mechanism underlying the regulation of PIK3R2 expression by H4K12 lactylation. This regulation, in turn, activates the PI3K/Akt/mTOR-signaling pathway and the fibrotic process, thus providing novel potential epigenetic targets for the prevention and treatment of lung fibrosis.
Adult mammals exhibit a limited capacity for tissue regeneration following injury and typically heal through scar formation. Mesenchymal stem/stroma cells (MSCs), which are phenotypically plastic and ubiquitous across tissues, play a critical role in maintaining tissue architecture during repair. We hypothesized that early events in regenerative and non-regenerative repair involve changes in MSC heterogeneity, which in turn determine repair outcomes. To test this hypothesis, we performed extensive single-cell RNA sequencing (scRNA-seq) in a mouse model of tissue injury. This model standardizes the comparison of non-regenerative and regenerative repair in adults with identical developmental stages and genetic backgrounds. Our analysis of MSCs during the early phases of tissue repair in adult mammals enabled the identification of distinct regenerative and non-regenerative MSC clusters, suggesting that specific MSC states may actively drive tissue repair outcomes. Furthermore, unsupervised approaches allowed us to revisit the functional signatures of MSCs centering on their impact on tissue structure (S), inflammation/immunity (I) and metabolism (M). By integrating these S, I and M functions, the SIM framework provides a conceptual model to interpret MSC behavior as a coordinated tissue-level response rather than a collection of isolated pleiotropic activities. This work positions MSCs at the center of the "SIM" triad, underscoring their pivotal role in tissue repair.
In our previous publications, we reported on esterifications carried out in a circulating flow MW system. It was found that circulation enables direct insight into the reactions from both chemical and energy perspectives. We examined productivity and scalability in terms of substrate, temperature, flow rate, and catalyst, and also analyzed the processes from the point of view of energy. Compared to the conventional linear setup, the circulation resulted in a significantly higher reaction rate, and thus a higher productivity, due to a drastic increase in flow rate. However, it also became clear that higher flow rates impose a substantially higher load on both the MW equipment and the overall system, which may easily lead to pipe perforations or instantaneous seal wear due to the higher operating pressure generated by the back pressure regulator (BPR) control unit. As higher pressures are associated with elevated temperatures, this raises considerable safety concerns. In this article, we aimed to eliminate the BPR unit. While this approach sacrifices a key advantage typically associated with the MW technology by restricting the operation to atmospheric pressure, the capabilities offered by the circulation solution may compensate for the inherent limitations of the new system, and, in turn, the safety risk may be reduced.
Dendritic cells (DCs) can activate T cells to trigger sustained antitumor immune responses, a process in which the continuous migration of antigen-loaded DCs from the tumor microenvironment to tumor-draining lymph nodes is critical. DCs have evolved a complex and dynamic regulatory network to mediate their migration to specific locations, with multiple transporters including amino acid transporters reported to participate in this process. Slc1a2 is highly expressed in the nervous system, where it mediates the clearance of extracellular glutamate, primarily in astrocytes; however, its role in the immune system remains unclear. In this study, we showed that activated DCs upregulated Slc1a2 to boost glutamate uptake, which in turn promoted DC functionality and antitumor vaccine potency. Furthermore, glutamate signaling induced Sema3A, which elevated small GTPase signaling pathways (e.g., RhoA/Rac1/Cdc42) and drove dynamic cytoskeletal remodeling, thereby providing the necessary molecular machinery for DC migration during antitumor immunity in vivo. We first demonstrated that Slc1a2-mediated glutamate metabolism functioned as a metabolic checkpoint for DC migration and antigen-specific immunity in vivo, with the Sema3A/small GTPase axis serving as the core mechanism linking glutamate signaling to cytoskeletal reorganization.
Chronic hepatitis B virus (HBV) infection remains a major global public health challenge, partly because of significant gaps in our understanding of its pathogenesis. Host factors play critical roles in regulating specific stages of the HBV life cycle, including replication. ZBTB7A, a member of the POK family of zinc-finger transcription factors, is involved in carcinogenesis and the modulation of certain viral infections. However, its role in HBV replication remains unclear, and this study aims to address this. We observed that ZBTB7A mRNA levels were higher in the immune-control phase than in the immune-tolerant phase and were inversely correlated with serum HBV DNA levels. ZBTB7A mRNA and protein levels were significantly downregulated in liver tissues from patients with chronic HBV infection, as well as in HBV-expressing hepatoma cells and primary human hepatocytes (PHHs). ZBTB7A overexpression markedly reduced HBV transcription and replication in HBV-infected hepatoma cells, PHHs, and an HBV-persistent mouse model. Mechanistically, ZBTB7A significantly repressed HBV transcription by suppressing viral core promoter activity, which in turn was achieved by reducing FXRα expression. ZBTB7A also physically interacted with FXRα, contributing to the inhibition of HBV transcription. In addition, ZBTB7A upregulated APOBEC3A expression, a mechanism that likely contributed to the observed reduction in covalently closed circular DNA (cccDNA) levels. Collectively, our data demonstrate that ZBTB7A suppresses HBV transcription and replication by diminishing both the amount and transcriptional activity of cccDNA. These findings suggest that ZBTB7A is a promising novel host-directed target for therapeutic intervention against HBV infection.
Stroke is a cerebrovascular condition that has emerged as the major cause of death globally, among other non-communicable diseases, due to its pathophysiological complexity and varied etiologies. Ischemic stroke (IS) represents the most prevalent subtype accounting for more than 70% of all stroke cases worldwide. The gut microbiome is considered the most abundant pool of microorganisms, which release metabolic products that facilitate gut-brain signaling. Despite advancements, the precise processes by which gut microbes influence stroke pathogenesis are not entirely understood. This review explores the immunomodulatory impact of gut-derived metabolites in IS, with a focus on short-chain fatty acids (SCFAs) and other bioactive substances. A comprehensive literature search was undertaken using MeSH terms such as gut microbiome, ischemic stroke, cerebral ischemia, brain infarction, intestinal flora, and gut-brain axis. We address how metabolites such as short-chain fatty acids produced by gut bacteria influence the host's immune system, which in turn affects the activity of neural networks linked to atherosclerosis, a major underlying mechanism of IS. Furthermore, this review also looks at new research opportunities and highlights evidence from studies conducted on humans and animals.
Maternal depression has been consistently linked to child ADHD symptoms, though much of this research is cross-sectional, single informant, and focused on prenatal or early postnatal depression. Consequently, the direction and temporality of this association is poorly understood. We used Random-Intercepts, Cross-Lagged Panel models (RI-CLPM) to examine bidirectional and transactional associations between maternal depression and child ADHD across pre-adolescence. Mothers and children (n = 1,806; 52% male, 77% White) participated across four waves during the COVID-19 pandemic (ages 10-13). At each timepoint, mothers self-reported depressive symptoms and children self-reported hyperactivity and inattention symptoms. Mothers also reported children's symptoms at the latter three waves. Parallel RI-CLPMs were estimated separately by informant and symptom dimension. Across both informants, maternal depression was associated with child hyperactivity and inattention at the between-dyad level (correlations ranging from .27 to .39). The clearest evidence of within-dyad association emerged for child-reported hyperactivity: higher child-reported hyperactivity at age 10 predicted higher maternal depressive symptoms at age 11, which in turn predicted higher child hyperactivity at age 12. Indirect effect analyses supported this transactional pathway. Maternal-reported models provided more limited but broadly convergent evidence for within-dyad effects on hyperactivity. Within-dyad effects were not observed for maternal- or child-reported inattention. Sociodemographic factors did not moderate cross-lagged parameters. Findings support transactional models of child development, with reciprocal associations between maternal depressive symptoms and child ADHD symptoms appearing most evident for hyperactivity. However, substantial overlap across ADHD symptom dimensions warrants caution in drawing strong conclusions about symptom-specific differences.
Pulmonary arterial hypertension (PAH) is a hemodynamic disorder that can progress to right heart failure and result in death. This study investigated the molecular mechanisms underlying the onset and progression of PAH to identify potential therapeutic targets. Peripheral blood samples from PAH patients were analyzed to assess serum levels of DKK1 and CKAP4, as well as NF-κB pathway activation. Supernatants from hypoxia-treated pulmonary artery endothelial cells (PAECs), plasmid-transfected cells, and SC75741-treated cells were used to modulate pulmonary artery smooth muscle cells (PASMCs). RT-qPCR, Western blot, and ELISA were employed to quantify DKK1 and CKAP4 expression and evaluate NF-κB pathway activation in PASMCs. EdU staining and CCK-8 viability assay were performed to assess cell proliferation, while DCFH-DA staining and ELISA were used to measure ROS, SOD, and MDA levels. DKK1 and CKAP4 expression were positively correlated, and both were upregulated with increasing pulmonary artery systolic pressure (PASP) in PAH patients. The supernatant from hypoxia-exposed PAECs induced NF-κB pathway activation, cell proliferation, and oxidative stress in PASMCs, effects that were inhibited by siDKK1, siCKAP4, and SC75741. Hypoxia stimulated PAECs to secrete DKK1, which in turn upregulated CKAP4 expression and activated the NF-κB pathway in PASMCs, promoting cell proliferation and oxidative stress.
The perioperative period is a critical and acute phase during which host-microbiota interactions play an essential role in determining susceptibility to anesthetic exposure and post-surgical stress. Immune homeostasis, gut barrier integrity, and metabolic regulation, as well as gut-brain and gut-liver axis, are highly dependent on the gut microbiota. However, this microbial ecosystem is disrupted during the perioperative period due to the combined effects of fasting, bowel preparation, surgical stress, hemodynamic alterations, and antibiotic and opioid use. Growing evidence has linked perioperative dysbiosis to a wide range of adverse outcomes, including infectious complications, anastomotic leakage, postoperative ileus, organ dysfunction, and perioperative neurocognitive disorders. Meanwhile, anesthetic and analgesic agents do not act in isolation from this ecosystem; rather, they engage in a bidirectional chemical interaction with the microbiota. These interactions can alter microbial structure and metabolite profiles, thereby influencing host metabolic processes, while microbial activity, in turn, affects drug disposition, immune response, and neuroinflammation. In this review, we first describe the vulnerability of gut microbiota homeostasis during the perioperative period and its associated clinical consequences. We subsequently elaborate on the mechanistic framework of anesthetic-microbiota crosstalk, highlighting pathways involving vascular, epithelial, immune and neural signaling. Then, we summarize emerging evidence demonstrating that different anesthetic and analgesic regimens generate discrete microbiota-metabolite signatures, which may underlie intersubject differences in postoperative recovery trajectories. Finally, we describe perioperative microbiota-targeted strategies, including probiotics and synbiotics, postbiotics and microbial consortia, nutritional optimization, and microbiome-based customization of anesthetic and analgesic protocols. Collectively, existing data suggest that preserving and actively modulating gut microbiota homeostasis represents a promising yet underexplored strategy for improving the safety of anesthesia and postoperative outcomes.
Tuberculosis (TB), caused by Mycobacterium tuberculosis (Mtb), is associated with clinical diversity and outcomes ranging from latent TB to active TB with distinct pathophysiologies. However, our understanding of the innate immune mechanisms related to the protection or progression of TB is limited. Among innate immune cells, the role of neutrophils is not fully elucidated, as they have been shown to exhibit both protective and harmful capacities in TB. We hypothesized that Mtb infection induces changes in neutrophil phenotype and function, influencing the infection outcomes. Based on clinical, bacteriological, and positron emission tomography with x-ray computed tomography (PET/CT) scan parameters, cynomolgus macaques infected by Mtb were stratified into two categories: animals that rapidly progressed to an active form of TB, designated as "fast progressors," and "slow progressors," which include low symptomatic or asymptomatic animals. In this study, we identified transcriptomic signatures of type I interferons and neutrophil degranulation in macaques with fast progression to active TB, which were not observed in animals with slow TB progression. Unsupervised mass cytometry analysis showed the emergence of blood immature neutrophils (CD101+ CD10-) in fast-progressing animals. In addition, bulk blood neutrophils from infected animals displayed capacities to modulate TNF-α production and cytotoxic function of CD8 T cells in a contact-dependent mechanism. In lung granulomas, neutrophils showed a tissue-specific phenotype (CD101- CD10+), with greater infiltration in animals with active TB. These data suggest that neutrophil subpopulations are associated with disease progression, with capacities to modulate CD8 T cells' functions, which in turn may contribute to disease progression.
The incorporation of phosphonate moieties into azaarenes via C-H functionalization is of paramount importance in pharmaceutical research, yet it remains a formidable synthetic challenge. Herein, we report an intermolecular cyclizative rearrangement approach for the efficient site-selective methylenephosphonylation of diverse azaarenes. This reaction proceeds via an orchestrated cascade sequence comprising regioselective 1,3-dipolar cycloaddition, [3,5]-sigmatropic rearrangement, and selective C-C bond cleavages. As a Lewis acid catalyst, ZnBr2 is crucial to this transformation: it not only delivers high conversion but also controls the regioselectivity of the initial cyclization, which in turn dictates the chemoselectivity of the entire process. This protocol is applicable to late-stage functionalization of various pharmaceuticals and ligands and facilitates further product diversification for advanced uses. This reaction represents the first example of the C-H methylenephosphonylation of arenes.
Microviscosity is an underexploited pathological cue that can be harnessed to regulate excited-state energy dissipation in organic near-infrared II (NIR-II, 1000-1700 nm) fluorophores. Here we introduce a twisted intramolecular charge transfer molecular rotor that uses microviscosity to gate radiative versus nonradiative decay, delivering a fluorescence "off" state in low-viscosity environments and a weak-but-sufficient NIR-II fluorescence turn-on in viscosity-elevated pathology while keeping nonradiative dissipation dominant for photothermal heating. A tunable donor-π-acceptor library was synthesized to drive absorption/emission toward the NIR-II window and calibrate a "dim-but-hot" photophysical profile. Molecularly dispersed FMR-1105-PEG enables in vivo NIR-II imaging in nonalcoholic fatty liver disease and acetaminophen-induced liver injury models. For tumor translation, micellization retains a measurable viscosity dependence (Förster-Hoffmann-type) while markedly increasing the apparent molar extinction coefficient, thereby enabling efficient 1064 nm photothermal activation, immunogenic cell death hallmarks in vitro, and imaging-guided tumor ablation even under tissue coverage, with favorable biosafety.