Introduction : With the growing expansion of participatory approaches in health, an increasing number of training initiatives have been developed to support these practices (Tourette-Turgis et al., 2019). However, these initiatives remain heterogeneous in their formats, objectives, and target audiences, reflecting a lack of consensus on what constitutes participatory approaches in practice (Paulo et al., 2023). Purpose of the research : This study aims to explore French-speaking training programs that foster collaboration among researchers, health professionals, and citizens. It focuses in particular on the underlying processes of acculturation, especially the ways in which citizens are introduced to research norms and the functioning of the healthcare system. Drawing on a review of grey literature and scientific studies on the topic, the study identified a wide range of training initiatives designed to promote the involvement of diverse groups in health research projects. Results : These programs most commonly seek to equip citizens and patients with the skills needed to participate in collaborative research. They vary according to the type of partnership (clinical, project, or educational) and the expected level of engagement, ranging from online modules to professional degree programs. Few initiatives, however, include specific training for health professionals. Conclusions : Training represents a key lever for supporting collaboration by fostering mutual understanding, adjustment, and recognition. However the imbalance in training opportunities raises questions about power relations, the professionalization of patients, and the epistemological limits of certain participatory approaches, highlighting critical challenges for the co-construction of knowledge in health. Introduction : Dans un contexte d’essor des démarches participatives en santé, les formations se multiplient pour accompagner ces pratiques (Tourette-Turgis et al., 2019). Ces initiatives sont toutefois marquées par une hétérogénéité de formats, d’objectifs et de publics, témoignant d’un manque de consensus concernant les démarches participatives en place (Paulo et al., 2023). But de l’étude : Cette étude vise à explorer les dispositifs de formation francophones qui soutiennent les collaborations entre chercheurs, professionnels de santé et citoyens. Elle interroge en particulier les logiques d’acculturation sous-jacentes, surtout au sujet de l’acculturation des citoyens aux normes de la recherche et au fonctionnement du système de santé. En s’appuyant sur une revue de littérature grise et d’études scientifiques sur le sujet, une diversité de formations a été identifiée, conçue pour favoriser l’implication de publics variés dans les projets de recherche en santé. Résultats : Ces dispositifs visent le plus souvent à doter les citoyens et patients de compétences leur permettant d’intégrer des projets collaboratifs. Ils varient selon le type de partenariat (clinique, recherche, formation) et le niveau d’implication attendu, allant de modules en ligne à des diplômes professionnalisants. Peu intègrent une formation des professionnels. Conclusions : La formation constitue un levier essentiel pour soutenir ces collaborations, en favorisant compréhension, ajustement et reconnaissance mutuelle. Toutefois le déséquilibre des trajectoires de formation questionne les rapports de pouvoir, la professionnalisation des patients et les limites épistémologiques de certaines démarches, soulevant des enjeux cruciaux pour la co-construction des savoirs en santé.
This paper investigates the affine formation maneuver control problem for underactuated multi-autonomous surface vehicle systems subject to limited sensing of neighbor state information and unknown disturbances. First, to address the situation where neighbor velocities are unavailable, a distributed super-twisting sliding mode observer is designed and used to estimate the unavailable velocities. Second, an observer-based affine formation controller is developed, which enables the formation to execute affine maneuvers, such as scaling, rotation, and translation, to navigate through complex environments. Moreover, a radial basis function neural network with an adaptive weight update law is employed to compensate for unknown disturbances. Then, the stability of the observer and the closed-loop system is analyzed, guaranteeing that all tracking errors and the estimation errors of the neural network are uniformly ultimately bounded. Finally, numerical simulations are conducted in a scenario with surface obstacles and disturbances to verify the effectiveness of the proposed observer and controller. The simulation results indicate that the proposed strategy achieves a mean absolute error of 0.310, a root mean square error of 1.201, and an integral absolute error of 21.668 for formation tracking accuracy.
The aggregation of the tau protein into intraneuronal fibrillar tangles is closely associated with the pathology of Alzheimer's disease. The endogenous defense system against this process includes molecular chaperones, among which DNAJB6b has emerged as a key component. Using a tau model system comprising the tau fragment 304-380C322S, which spans the amyloidogenic core of ex vivo Alzheimer's disease fibrils, we investigated the impact of DNAJB6b on tau fibril formation. Here, we show that DNAJB6b potently delays tau aggregation by co-assembling with small tau aggregates and by binding to mature fibrils, thereby reducing their ability to catalyze further fibril growth. This interplay between tau and the chaperone results in greatly reduced fibril formation rate and a lower final fibril mass, which we interpret as increased tau solubility. Moreover, solution-state NMR spectroscopy confirms that DNAJB6b does not interact with tau monomers.
Ultrasound-guided regional anesthesia (UGRA) requires knowledge of anatomy, hand-eye coordination, and safe needling techniques. Extended reality (XR), including augmented reality, virtual reality (VR), and mixed reality, is an emerging simulation modality for UGRA training. We sought to conduct a systematic review to evaluate the current evidence on the effectiveness of XR for UGRA training. We searched MEDLINE, Embase®, the Cochrane Library, Web of Science®, and the Cumulative Index to Nursing and Allied Health Literature (CINAHL) from inception to 15 November 2024. We included randomized controlled trials, observational studies, and case series. We categorized outcomes by Kirkpatrick levels and assessed risk of bias. From 1,703 articles, we included seven studies (N = 137 participants). Four studies reported participant reactions. One trial (n = 29) found higher self-confidence in anatomical landmark identification and procedural performance (P < 0.001). One study assessed knowledge, reporting a score increase (P < 0.001). Augmented reality head-mounted display studies found shorter procedure times (10 vs 7 sec; P = 0.03 and 18 vs 12 sec; P = 0.002), higher needle visibility (34% vs 57%; P < 0.001 and 20% vs 38%; P = 0.001), and fewer head movements (P = 0.02). Immersive VR trials found no between-group differences in ratings or error scores (mean difference, -3.3; 95% confidence interval, -12.7 to 6.1; P = 0.51). The only randomized controlled trial was a pilot study that was terminated early and was underpowered for outcomes. We found substantial variability in XR definition, study design, methodologic rigour, and study quality. The current evidence does not conclusively support widespread adoption of XR for UGRA training, owing to heterogeneity, small samples, and underpowered studies. Currently, XR remains at an early and experimental stage in development. PROSPERO ( CRD42023424194 ); first submitted 18 May 2023. RéSUMé: OBJECTIF: L’anesthésie locorégionale échoguidée requiert des connaissances en anatomie, de la coordination main-œil et la maîtrise de techniques d’insertion d’aiguille sécuritaires. La réalité étendue, qui englobe la réalité augmentée, la réalité virtuelle (RV) et la réalité mixte, constitue une modalité de simulation émergente pour la formation en anesthésie locorégionale échoguidée. Nous avons cherché à mener une revue systématique afin d'évaluer les données probantes actuelles sur l’efficacité de la réalité étendue dans la formation dans ce domaine. Nous avons effectué des recherches dans MEDLINE, Embase®, la bibliothèque Cochrane, Web of Science® et le répertoire CINAHL (Cumulative Index to Nursing and Allied Health Literature), depuis leur création jusqu’au 15 novembre 2024. Nous avons inclus des études randomisées contrôlées, des études observationnelles et des séries de cas. Nous avons classé les critères d'évaluation selon les niveaux de Kirkpatrick et évalué le risque de biais. Sur 1703 articles recensés, sept études ont été retenues (n = 137 personnes). Quatre études faisaient état des réactions des personnes participantes. Une étude (n = 29) a relevé une plus grande confiance en soi dans l’identification des repères anatomiques et dans l’exécution des gestes (P < 0,001). Une étude a évalué les connaissances, rapportant une d’une amélioration du score (P < 0,001). Les études sur les afficheurs tête-haute en réalité augmentée ont mis en évidence des durées d’intervention plus courtes (10 vs 7 s; P = 0,03 et 18 vs 12 s; P = 0,002), une meilleure visibilité de l’aiguille (34 % vs 57 %; P < 0,001 et 20 % vs 38 %; P = 0,001) et moins de mouvements de tête (P = 0,02). Les essais de RV immersive n’ont révélé aucune différence entre les groupes quant aux scores d'évaluation ou d’erreur (différence moyenne, −3,3; intervalle de confiance à 95 %, −12,7 à 6,1; P = 0,51). Le seul essai randomisé contrôlé était une étude pilote interrompue prématurément et dont la puissance était insuffisante pour évaluer ces critères. Nous avons constaté une variabilité importante dans la définition de la réalité étendue, la conception des études, la rigueur méthodologique et la qualité des études. Les données actuelles ne permettent pas de conclure en faveur d’une adoption généralisée de la réalité étendue dans la formation en anesthésie locorégionale échoguidée, en raison de l’hétérogénéité des études, de la petite taille des échantillons et de la puissance insuffisante. À ce jour, la réalité augmentée en est encore à un stade précoce et expérimental de développement. ENREGISTREMENT DE L’éTUDE: PROSPERO ( CRD42023424194 ); première soumission le 18 mai 2023.
Myxococcus xanthus is a Gram-negative bacterium with a complex life cycle that includes vegetative swarming and fruiting-body formation. Previous studies have shown that the exopolysaccharides (EPS) produced by M. xanthus was essential for the developmental process, but the information concerning EPS structure in M. xanthus was lacking. In this study, a novel soluble polysaccharide EPS15M was purified from the extracellular supernatant of M. xanthus DK1622, which was mainly composed of mannose, glucose, glucosamine, and rhamnose. Methylation analysis suggested the prevalence of terminal and 2-linked glycosyl residues, consistent with a branched structure. NMR and liquid chromatography-mass spectrometry analyses confirmed that EPS15M is an α-D-(1,2)-linked mannan backbone with α-D-(1,6)-linked mannose/glucose branches, a structure that differs from any extracellular polysaccharide reported to date from myxobacteria. Besides, EPS15M modestly improves the developmental sporulation of the csgA mutant (approximately 17% of the wild‑type level) at a concentration of 0.1 mg/mL. Collectively, our findings present the first high-resolution structure of a soluble exopolysaccharide from myxobacteria and demonstrate its partial functional compensation of csgA mutant, suggesting that EPS15M may be involved in C‑signal‑mediated developmental sporulation. These results open new avenues for understanding the regulatory role of polysaccharides in the myxobacterial life cycle.
With the rapid rate of industrial modernization, the continuous emissions of carbon dioxide (CO2) are greatly disturbing the natural carbon balance and contributing to global warming. To mitigate this effect, photocatalytic reduction of carbon dioxide has gained popularity as a means of converting it into useful products. This study examined the photocatalytic reduction of aqueous CO2 under ultraviolet irradiation (UV) using green carbon dots (CDs) made from pomegranate peel, nano titanium dioxide (TiO2), TiO2/CDs composite, and metal-loaded carbon dots-modified titanium oxide (Cu-CDs/TiO2). The reduction of CO2 gas in water was carried out in a batch reactor utilizing the four photocatalysts. In order to verify the catalysts' functional groups, nanoscale shape, and composite production, FTIR and TEM were used for characterization. The findings demonstrated that oxygenated products were preferred by pure CDs, but the performance of TiO2 was very pH dependent, with ester formation being promoted in acidic environments and CO formation in alkaline ones. An increase in CO selectivity resulted from the TiO2-CDs composite's enhanced charge separation and electron transfer; a change toward decreased hydrocarbons in product distribution was brought about by the addition of Cu, which opened hydrogenation pathways.
Antibiotic resistance is a global health threat, driving the need for new molecules that kill bacteria via nontraditional mechanisms. Here, we present a computational de novo design strategy for α-helical peptides that self-assemble into large, stable and membrane-spanning nanopores with antimicrobial activity, including in vivo efficacy against drug-resistant pathogens. Molecular dynamics simulations guided the selection of sequences for transmembrane barrel-stave pore formation, which were validated by microscopy, electrophysiology and fluorescence assays. Using computational and experimental analyses, including negative design controls, we developed general design guidelines and 52 modular sequence templates with tunable antimicrobial, pore-forming properties. Mechanistic studies confirmed bacterial cytoplasmic membrane disruption via designed nanopore formation. A tuned lead peptide selectively killed drug-resistant ESKAPEE bacteria, including Acinetobacter baumannii, without harming human cells, and showed anti-infective efficacy in preclinical mouse infection models. The framework presented here enables the design of synthetic peptide nanopores for precision antimicrobials, anticancer agents, molecular sensors and delivery systems.
The bradykinin 2 receptor (B2R) is one of two members of the kinin receptor family and is a G protein-coupled receptor (GPCR) that regulates important physiological processes including pain, inflammation, and cardiovascular homeostasis. Functionally, B2R is activated by the kinin peptides, bradykinin and kallidin. Carboxypeptidase cleavage of bradykinin and kallidin results in the formation of des-Arg9-bradykinin (DABK) and des-Arg10-kallidin (DAKD) respectively, which are classically described as agonists of the bradykinin 1 receptor (B1R), the other member of the kinin family. Affinity binding studies have reported that DABK and DAKD can also bind to B2R. Upon activation, B2R signals through the recruitment of heterotrimeric G proteins, and earlier work has highlighted the promiscuous G protein coupling to B2R in response to bradykinin. However, a comprehensive G protein activation profile of the B2R receptor when activated by the other endogenous agonists has not yet been established. In this study, we used Bioluminescence Resonance Energy Transfer (BRET)-based in vitro assays to monitor the coupling of 14 different Gα proteins upon B2R activation. Our results show that there is a shift in G protein activation profiles and kinetics among the different peptides. By analyzing responses across a range of agonist concentrations, we further identified biased signaling among these peptides, with differential activation of specific Gα subtypes. These results improve understanding of intracellular mechanisms through which the kinin system regulates different aspects of its physiological role and provide valuable information for the design of drugs targeting the kinin receptors.
This study examines a nanosuspension of secondary metabolites (NSM) derived from Streptomyces chrestomyceticus strain ADP4, previously reported for its antifungal efficacy against Candida spp., and extends its evaluation to WHO-listed high-priority bacterial pathogens. The antibacterial potential of the NSM was assessed against Staphylococcus aureus ATCC 25,923, Enterococcus faecium ATCC 49,924, and Enterococcus faecalis ATCC 29,212. Transmission electron microscopy revealed the formation of uniformly dispersed, spherical nanoparticles with an average diameter of 14 ± 4.26 nm. Compared with the crude secondary metabolites (SMs), the NSM demonstrated noticeably enhanced antibacterial performance, reflected by increased zones of inhibition (10%, 16%, and 26%), significant reductions in minimum inhibitory concentrations (53%, 42%, and 71%), and improved minimum biofilm inhibitory concentrations (41%, 30%, and 70%) against S. aureus, E. faecium, and E. faecalis, respectively. The observed enhancement in antimicrobial and antibiofilm activity highlights the role of nano-formulation in improving the bioavailability and biological efficiency of microbial SMs. Also, the flow cytometric analysis of bacterial cells treated with SMs and NSM revealed an increase in membrane permeability in a dose- and time-dependent manner. These findings highlight the potential of actinobacteria-derived NSM as environmentally relevant and support its applicability in macromolecular and surface-associated biomedical applications, including antibiofilm coatings for medical devices, wound dressings, and infection-resistant biomaterials making NSM as a potential strategy for sustainable and microbe-driven innovation.
In the central nervous system, Ca2⁺ signaling plays a pivotal role in various cellular processes, including neuronal development and maturation. Disruption of Ca2⁺ homeostasis impairs auditory circuit formation, compromising auditory processing and behavior. Here, we aimed to investigate systematically the development of Ca2⁺ signaling in neurons of the medial nucleus of the trapezoid body (MNTB), a brainstem nucleus critical for sound localization. Using Ca2⁺ imaging in brain slices from mice genetically expressing the Ca2⁺ indicator GCaMP in glycinergic cells, we studied Ca2⁺ signaling in MNTB neurons from mice of either sex at three developmental milestone stages, in response to both bath-applied agonists of various neurotransmitter receptors and synaptic activation of glutamatergic afferents. Prior to hearing onset at postnatal 7 days (P7), robust Ca2⁺ responses were evoked in MNTB neurons upon activation of glutamate receptors (NMDARs, AMPARs, and group I (Gp-I) mGluRs by 200 μM NMDA, 100 μM AMPA, and 200 μM 3,5-DHPG, respectively), whereas after hearing onset (P14 and P21), responses induced by NMDA and 3,5-DHPG declined markedly while AMPA-induced responses remained relatively strong. Ca2⁺ responses upon application of GABA (100 μM) and glycine (200 μM) were detected in neonatal mice, diminished in a few days after birth, and almost completely disappeared by P7. Whole-cell patch-clamp recordings showed that stimulation of excitatory afferents evoked action potentials across all ages with no differences in firing frequency up to 100 Hz, but Ca2⁺ responses varied in a stimulus intensity- and frequency-dependent manner and exhibited developmental downregulation. Ca2⁺ signaling induced by activation of the major transmitter receptors in MNTB neurons is highly developmentally down-regulated.
Fungi frequently encounter nutrient limitation, osmotic stress, oxidative pressure, light fluctuations, temperature shifts, and host-associated stresses. Rather than responding passively to deteriorating environments, many fungi actively redirect development from vegetative growth toward asexual sporulation, sexual reproduction, or resting structure formation. These transitions promote dispersal, dormancy, stress resistance, or genetic diversification, but they are not governed by a single conserved stress-response pathway. Instead, conserved nutrient- and stress-sensing modules are integrated with lineage-specific developmental circuits. This review synthesizes recent advances in understanding how environmental stress, particularly nutrient limitation, regulates fungal reproductive development. We discuss nutrient limitation as both a physiological constraint and a developmental signal, and compare regulatory mechanisms across yeasts, filamentous ascomycetes, and pathogenic fungi. Major pathways and regulatory layers include cAMP-PKA, TOR, HOG MAPK signaling, light-responsive systems, the Velvet complex, chromatin regulation, diffusible chemical signals, and sexual-stage-specific adenosine-to-inosine (A-to-I) mRNA editing. Together, these mechanisms determine whether fungi remain vegetative or commit to reproduction. We also evaluate why sexual reproduction may be favored under stressful or low-fitness conditions. While asexual spores and resting structures can enhance survival and dispersal, costly meiotic reproduction may be promoted when recombination allows offspring or alleles to escape maladapted genetic backgrounds. Fitness-associated sex and abandon-ship models therefore provide useful evolutionary frameworks for interpreting stress-induced reproduction. Understanding these mechanisms is timely and important because fungal reproductive switching affects industrial spore production, pathogen transmission, disease management, and evolutionary potential. Future progress will require causal, ecologically grounded models linking environmental perception, molecular regulation, reproductive output, and fitness consequences.
In this study, silver nanoparticles were green synthesized using Cotoneaster lacteus and characterized for the first time in a comprehensive way. The phytochemical composition of the extract was analyzed by GC-MS/MS and LC-ESI-MS/MS, and the main constituents were identified as linoleic acid (63.56%) and trans-ferulic acid (55.94 ng/mL). The C. lacteus extract-based AgNPs were characterized using UV-Vis, FT-IR, XRD, SEM-EDX, STEM, and zeta potential analyses. XRD results showed a face-centered cubic crystal structure with an average crystallite size of 25.42 nm, while STEM images indicated predominantly spherical particles with an average size of 20.75 ± 3.79 nm. In the biological assays, C. lacteus extract-based AgNPs showed noticeably higher antibacterial activity compared to both the crude extract and AgNO₃. Antibiofilm inhibition rates also exceeded 50%. Similarly, antioxidant assays (DPPH˙, ABTS˙⁺, and FRAP) confirmed stronger radical scavenging and reducing ability of the AgNPs compared to the extract. DFT calculations suggest that trans-ferulic acid and hesperidin interact with silver atoms through oxygen-containing functional groups, which likely contribute to nanoparticle formation and stabilization. Molecular docking results further showed favorable interactions between ferulic acid, hesperidin, and Ag complexes with key antioxidant and antibacterial target proteins, supporting the experimental findings. Overall, plant-mediated AgNPs appear promising for biomedical and biotechnological applications due to their antibacterial, antibiofilm, and antioxidant properties.
Synthetic riboswitches provide protein-independent, modular control of gene expression, yet selecting aptamers that reliably couple ligand binding to regulatory switching remains challenging. Here, we identify and mechanistically characterise G12, a doxycycline-binding aptamer with remarkably high regulatory performance in yeast and human cells. We provide evidence that RNA Capture-SELEX efficiently enriches aptamers with ligand-responsive conformational switching. We compared conventional SELEX and RNA Capture-SELEX using the same starting library followed by NGS analysis and in vivo screening, which led to the identification of G12. G12 binds doxycycline with low-nanomolar affinity and strict discrimination against close derivatives, thus enabling high-dynamic-range riboswitch control of translation in yeast and splicing in human cells. Single-molecule force spectroscopy with optical tweezers revealed that doxycycline stabilises a folding intermediate independent of the closing stem P1, which primarily acts as a scaffold for correct aptamer folding. Mutational analysis and chemical probing identified tertiary contacts between loops L2 and L3 in this intermediate state. Stopped-flow fluorescence spectroscopy further supported a two-step binding mechanism consistent with efficient regulatory switching. Together, these findings deepen our understanding of regulatory aptamer selection and function and expand the synthetic biology toolbox with a high-performance doxycycline-responsive riboswitch.
Post-radiation head and neck cancer (HNC) patients with xerostomia are predisposed to opportunistic oral candidiasis. While resistance to fluconazole, a first-line antifungal agent, is increasingly prevalent, it is not well-characterized among these patients. Thus, we aimed to evaluate fluconazole resistance and associated factors among Candida isolates from this population. Oral Candida isolates from xerostomic post-radiation HNC patients and healthy controls, with demographic, clinical, and biofilm-forming data, were obtained from previous studies. Fluconazole susceptibility was tested using E-test™ and interpreted according to CLSI-M60 guidelines. Data were analyzed using Chi-square test, Mann-Whitney-U test and multivariate regression with significance level at p < 0.05. Fluconazole resistance was observed in 8.8% of C.albicans and 16.7% of C.tropicalis isolates from HNC patients, but none from healthy controls, except for the intrinsically resistant C.krusei. The prevalence of fluconazole resistance was significantly higher in HNC group at both isolate and patient levels (p = 0.037 and 0.027, respectively). This association persisted after adjusted for potential confounders. Interestingly, fluconazole resistance was significantly associated with higher Candida load in cancer patients, while it was associated with lower biofilm formation in controls. Fluconazole resistance maybe more common in xerostomic post-radiation HNC patients, thus, cautions are warranted when prescribing fluconazole.
The colonization of the periodontal sulcus by pathogenic oral bacteria may be promoted by the eruption of permanent teeth. To analyse the evolution of the oral microbiota during teeth eruption. Sub-gingival microbiome analyses (16S) of primary teeth and eruption permanent teeth sulci (case-matched) were carried out with periodontitis sample controls. A number of disease-associated bacteria including Tannerella forsythia, Treponema denticola and Fretibacterium sp. HMT 360 have been detected in erupting teeth samples but were absent in primary teeth. The formation of deeper sulcus during permanent teeth eruption may create an early colonization niche for periodontal pathogens in older children or young adults which may be implicated later in periodontal diseases.
Objective: To investigate the effects and mechanisms of polyvinyl alcohol/ionic liquid-tannic acid composite hydrogel (PIT) on wound healing of full-thickness skin defects in diabetic mice. Methods: This study was an experimental research involving grouped design and repeated measurements. An ionic hydrogel matrix crosslinked by polyvinyl alcohol-4-(1H)-vinylimidazole-1-methylene benzoic acid and oxidized hyaluronic acid was prepared, and tannic acid was loaded via Cu2+ chelation to construct PIT. A 1,1-diphenyl-2-picrylhydrazyl (DPPH) solution was prepared and reacted respectively with tannic acid and PIT for 24 hours. An ultraviolet spectrophotometer was used to detect the DPPH radical scavenging rate. According to the random number table method (the same grouping method below), mouse macrophage RAW264.7 cells were divided into a phosphate buffered saline (PBS) group cultured with PBS, as well as a hydrogen peroxide group and a PIT group, in which cells were first treated with hydrogen peroxide for 12 hours and then respectively cultured under routine condition and with PIT. After 24 hours of culture, the fluorescent probe method was adopted to detect the intracellular reactive oxygen species (ROS) level. Escherichia coli ATCC 25922, Staphylococcus aureus ATCC 25923, methicillin-resistant Staphylococcus aureus (MRSA) BNCC 337371, and human umbilical vein endothelial cells (HUVECs) were collected and all divided into PBS group, tannic acid (TA) group, and PIT group, which were cultured with PBS, TA solution, and PIT, respectively. After 12 hours of bacterial culture, the plate counting method was used to count bacterial colonies; after 24 hours of cell culture, the tube formation assay was performed to measure the total tube length, the number of branching nodes, and the number of branches. The sample size of all the above experiments was 3. Eighteen 8-week-old male Kunming mice were selected and divided into PBS group, TA group, and PIT group (with 6 mice in each group) to establish a full-thickness skin defect wound model of diabetes (with one wound in each mouse). At post injury day (PID) 0 (immediately), the wounds of mice in PBS group, TA group, and PIT group were treated respectively with PBS, TA solution, and PIT by topical application, and then the dressings were changed daily. The wound healing status was observed at PID 0, 4, 8, and 12, and the wound healing rates at PID 4, 8, and 12 were calculated. At PID 12, wound tissue was harvested. Hematoxylin-eosin staining was performed to observe the status of wound re-epithelialization and to measure the thickness of newly formed epithelium. Masson staining was performed to observe the deposition of collagen fibers in wounds and to calculate the proportion of collagen fiber-positive area. Results: After 24 hours of reaction, the DPPH radical scavenging rate of PIT was significantly higher than that of TA (t=16.35, P<0.05). After 24 hours of culture, the ROS level of RAW264.7 cells in hydrogen peroxide group was significantly higher than that in PBS group (P<0.05), and the ROS level of RAW264.7 cells in PIT group was significantly lower than that in hydrogen peroxide group (P<0.05). After 12 hours of culture, the bacterial colony counts of Escherichia coli, Staphylococcus aureus, and MRSA in PIT group were significantly less than those in PBS group and TA group (P<0.05). After 24 hours of culture, compared with those in PBS group and TA group, the total tube length of HUVECs in PIT group was significantly increased (with P values both <0.05), and the number of branching nodes and the number of branches increased significantly (with P values all <0.05). From PID 0 to 12, the wounds of mice in all three groups healed gradually. At PID 4, 8, and 12, the wound healing rates of mice in PIT group were (31.6±2.0)%, (51.8±2.5)%, and (97.9±1.5)%, respectively, which were significantly higher than (18.6±0.6)%, (39.5±2.0)%, and (74.6±2.0)% in PBS group and (21.5±1.1)%, (40.7±0.8)%, and (85.3±2.1)% in TA group (P<0.05). At PID 12, the wound re-epithelialization of mice in PBS group was incomplete, and collagen fibers were sparsely distributed with disordered arrangement; the degree of wound re-epithelialization of mice in TA group was higher than that in PBS group, and collagen fibers were distributed in bundles with loose arrangement; the degree of wound re-epithelialization of mice in PIT group was higher than that in TA group, and collagen fibers were densely and orderly arranged in layers. At PID 12, compared with those in PBS group and TA group, the thickness of newly formed epithelium in wounds of mice in PIT group was significantly increased (with P values both <0.05), and the proportion of collagen fiber-positive area was significantly increased (with P values both <0.05). Conclusions: PIT significantly accelerates the wound healing of full-thickness skin defects in diabetic mice through multiple mechanisms driven by metal ion chelation including antibacterial, antioxidant, and pro-angiogenesis activities, thereby improving the quality of tissue repair. 目的： 探讨聚乙烯醇/离子液体-单宁酸复合水凝胶（PIT）对糖尿病小鼠全层皮肤缺损创面愈合的影响及其机制。 方法： 该研究为成组设计及重复测量设计实验研究。制备聚乙烯醇-4-（1H-乙烯基咪唑）-1-亚甲基苯甲酸与氧化透明质酸交联的离子水凝胶基质，通过铜离子螯合负载单宁酸，构建PIT。配制1，1-二苯基-2-三硝基苯肼（DPPH）溶液，分别与单宁酸和PIT反应24 h后，采用紫外分光光度计检测DPPH自由基清除率。取小鼠巨噬细胞RAW264.7，采用随机数字表法（分组方法下同）分为加入磷酸盐缓冲液（PBS）培养的PBS组，以及均经过氧化氢处理12 h后分别常规培养、加入PIT培养的过氧化氢组、PIT组，培养24 h后，采用荧光探针法检测细胞中活性氧水平。取大肠埃希菌ATCC 25922、金黄色葡萄球菌ATCC 25923、耐甲氧西林金黄色葡萄球菌（MRSA）BNCC 337371、人脐静脉内皮细胞（HUVEC），将每种材料分别分为PBS组、单宁酸组、PIT组，各组分别加入PBS、单宁酸溶液、PIT进行培养，细菌培养12 h后，采用平板计数法计数菌落；细胞培养24 h后，行细胞成管实验，测算成管总长度、分支节点数、分支数。前述实验样本数均为3。取18只8周龄雄性昆明小鼠，分为PBS组、单宁酸组、PIT组（每组6只），构建为糖尿病全层皮肤缺损创面模型（每只小鼠1个创面），伤后0 d（即刻），分别于PBS组、单宁酸组、PIT组小鼠创面滴加PBS、单宁酸溶液、PIT处理，之后每天换药。观察伤后0、4、8、12 d创面愈合情况，并计算伤后4、8、12 d创面愈合率。伤后12 d，取创面组织，行苏木精-伊红染色，观察创面上皮再生情况，并且测量新生上皮厚度；行Masson染色，观察创面胶原纤维沉积情况，并且计算胶原纤维阳性面积占比。 结果： 反应24 h后，PIT的DPPH自由基清除率显著高于单宁酸（t=16.35，P<0.05）。培养24 h后，过氧化氢组RAW264.7细胞中活性氧水平显著高于PBS组（P<0.05），PIT组RAW264.7细胞中活性氧水平显著低于过氧化氢组（P<0.05）。培养12 h后，PIT组大肠埃希菌、金黄色葡萄球菌、MRSA菌落数均显著少于PBS组及单宁酸组（P<0.05）。培养24 h后，与PBS组及单宁酸组比较，PIT组HUVEC成管总长度显著延长（P值均<0.05），分支节点数与分支数显著增多（P值均<0.05）。伤后0~12 d，3组小鼠创面均逐步愈合。伤后4、8、12 d，PIT组小鼠创面愈合率分别为（31.6±2.0）%、（51.8±2.5）%、（97.9±1.5）%，均显著高于PBS组的（18.6±0.6）%、（39.5±2.0）%、（74.6±2.0）%和单宁酸组的（21.5±1.1）%、（40.7±0.8）%、（85.3±2.1）%（P<0.05）。伤后12 d，PBS组小鼠创面再上皮化不完全，胶原纤维分布稀疏且排列紊乱；单宁酸组小鼠创面再上皮化程度高于PBS组，胶原纤维呈束状分布但排列仍较松散；PIT组小鼠创面再上皮化程度高于单宁酸组，胶原纤维呈致密有序的层状排列。伤后12 d，与PBS组及单宁酸组比较，PIT组小鼠创面新生上皮厚度显著增加（P值均<0.05），胶原纤维阳性面积占比显著增大（P值均<0.05）。 结论： PIT通过金属离子螯合驱动的抗菌、抗氧化、促血管生成多重机制，显著加速糖尿病小鼠全层皮肤缺损创面的愈合，并提高组织修复质量。.
Ankle clonus is a sustained, involuntary, rhythmic muscle contraction frequently observed in humans with spinal cord injury (SCI). Although its pathophysiology remains incompletely understood, converging evidence suggests a role for brainstem systems in its generation. Following SCI, brainstem neuromodulatory inputs partially compensate for the loss of descending motor pathways by regulating motoneuron excitability during involuntary contractions, suggesting their involvement in the generation of clonus. To test this hypothesis, motoneuron excitability in response to Ia synaptic input was quantified using the soleus H-reflex and maximal motor response (H/M ratio). Brainstem involvement was assessed using the long-latency component of the cutaneous reflex (LLR), which is influenced by monoaminergic pathways, and the StartReact response, in which reductions in reaction time to a loud acoustic stimulus are thought to reflect descending drive from the reticular formation, measured in the tibialis anterior and soleus muscles. We studied males and females with chronic SCI, both with and without ankle clonus, using standardized clinical tests across two days. Participants with clonus showed elevated H/M ratios, indicating increased motoneuron excitability, whereas those without clonus exhibited lower values than controls. Additionally, individuals with clonus exhibited longer LLR duration and greater LLR magnitude in both muscles, along with shorter reaction times to startle stimuli, consistent with enhanced monoaminergic and reticulospinal contributions. Notably, LLR duration was positively correlated with both StartReact response and H/M ratio. Together, these findings suggest a potential contribution of descending brainstem systems to the maintenance of clonus after chronic SCI.Significance Statement Ankle clonus is a repetitive, involuntary muscle contraction often seen after spinal cord injury (SCI). Its cause is not fully understood, but brainstem pathways may contribute by increasing the excitability of spinal motoneurons. In this study, we measured reflex responses in leg muscles and reactions to sensory and startling stimuli in people with chronic SCI, with and without clonus. Individuals with clonus showed stronger and longer reflex responses and faster reactions to startling sounds. These findings suggest that brainstem pathways involved in movement control may help maintain ankle clonus after SCI.
Objective: To evaluate the effects of pulsed dye laser (PDL) combined with fractional carbon dioxide laser in the treatment of hypertrophic scars (HSs) in children. Methods: This study was a meta-analysis. Databases including PubMed, Embase, Cochrane Library, Web of Science, China National Knowledge Infrastructure, Wanfang Data Knowledge Service Platform, and VIP Chinese Journal Service Platform were retrieved to obtain the publicly published retrospective and prospective studies on the effects of PDL combined with fractional carbon dioxide laser for pediatric HS from the establishment of each database to October 31, 2024. The outcome indexes included the total score of the patient and observer scar assessment scale (POSAS) and laser treatment-related adverse reactions. Stata version 16.0 software was used to conduct a meta-analysis. Results: A total of 7 studies were included, involving 348 children with HSs. Meta-analysis results showed that the total POSAS scores in HS children who received PDL combined with fractional carbon dioxide laser treatment were significantly lower than those before treatment (with a standardized mean difference of -7.76, a 95% CI of -10.91 to-4.61, P<0.05). Subgroup analysis results suggested that geographic distribution, scar duration (divided into ≤3 months group and >3 months group), intervention measures, and fractional carbon dioxide laser energy parameters might be sources of heterogeneity for the total POSAS score. The incidence of adverse reactions of PDL combined with fractional carbon dioxide laser for pediatric HS was 3.50% (with a 95% CI of 1.60% to 6.00%, P<0.05). There was no publication bias in the total POSAS score, its subgroup analyses (including geographic distribution, scar duration, intervention measures, fractional carbon dioxide laser energy parameters, and assessor), or adverse reactions related to laser treatment (P>0.05). Conclusions: PDL combined with fractional carbon dioxide laser is beneficial for the recovery of pediatric HS, with a low rate of adverse reactions and a favorable safety profile. For pediatric HS, the early intervention window within 3 months after scar formation should be prioritized, and a regimen combining low-energy fractional carbon dioxide laser flexibly with PDL should be adopted to achieve better therapeutic outcomes. 目的： 评价脉冲染料激光（PDL）联合点阵二氧化碳激光治疗儿童增生性瘢痕（HS）的效果。 方法： 该研究为荟萃分析。检索PubMed、Embase、Cochrane Library、Web of Science、中国知网、万方数据知识服务平台和维普中文期刊服务平台等数据库自建库至2024年10月31日公开发表的关于PDL联合点阵二氧化碳激光治疗儿童HS效果的回顾性研究和前瞻性研究，结局指标包括患者与观察者瘢痕评估量表（POSAS）总评分和激光治疗相关不良反应。采用Stata 16.0软件进行荟萃分析。 结果： 共纳入7篇文献348例HS患儿。荟萃分析结果显示，HS患儿采用PDL联合点阵二氧化碳激光治疗后POSAS总评分明显低于治疗前（标准化均数差为-7.76，95%CI为-10.91~-4.61，P<0.05）。亚组分析结果提示，地域分布、瘢痕病程（分为≤3个月组和>3个月组）、干预措施以及点阵二氧化碳激光能量参数可能是POSAS总评分的异质性来源。HS患儿采用PDL联合点阵二氧化碳激光治疗的不良反应发生率为3.50%（95%CI为1.60%~6.00%，P<0.05）。POSAS总评分及其各亚组分析（包括地域分布、瘢痕病程、干预措施、点阵二氧化碳激光能量参数、评估者）和激光治疗相关不良反应均不存在发表偏倚（P>0.05）。 结论： PDL联合点阵二氧化碳激光治疗有利于儿童HS的恢复，并且不良反应发生率较低，具有良好的安全性。针对儿童HS，应优先把握瘢痕形成后3个月及以内的早期干预窗口期，采用低能量点阵二氧化碳激光灵活搭配PDL方案，以取得更佳疗效。.
We present a high-speed force microscopy platform mounted on a confocal microscope with a z sample stage enabling long-range, high-speed force spectroscopy measurements on biological samples. The control software is built on a field-programmable gate array (FPGA)-based data acquisition and processing system, complemented with a custom graphical user insterface (GUI). We introduce smart algorithms based on probe-engagement prediction algorithms that leverage previously measured probe-sample contact to accelerate the probe engagement to mm/s and decelerate it in proximity of contact to user-defined μm/s velocities. This significantly reduces long-range force curve acquisition time and data density. Using this system, we provide proof-of-concept mechanical maps of cell clusters to extract topography and viscoelastic parameters. To further explore the versatility of our system, we probed the forces required to extract membrane tethers from monocytic cells using ultrashort cantilevers functionalised with adhesion molecules. Our system allowed us to cover retract velocities from ~1 μm/s up to ~6000 μm/s, and to explore the dynamics of tether formation at physiologically relevant velocities. Our results show that coupling extended z displacement with the prediction-based engagement algorithms enables rapid, quantitative mechanical mapping of heterogeneous biological samples with large topography variation and supports measurements requiring long distances at high velocities.
Amorphous solid dispersions (ASDs) have been shown to be an effective formulation strategy for improving apparent solubility, dissolution rate, and hence oral bioavailability of poorly aqueous soluble drugs. In this perspective, we review recent progress in understanding the role of polymer concentration, particularly the overlap concentration, c*, in governing crystallization of high drug loaded ASDs. The overlap concentration, c*, is a threshold above which adjacent polymer coils start to contact. Recent work has shown that when polymer concentration is below c*, the presence of the continuous polymer-free amorphous drug domain allows crystallization to proceed similarly to neat drug, resulting in limited inhibition. Above c*, formation of a homogeneous polymer rich matrix suppresses crystallization, primarily by delaying the first nucleation event. Representative case studies demonstrate the generality of this framework across diverse drug-polymer systems, although limitations arise for sufficiently high molecular weight polymers. Overall, the c*-guided rheological approach provides a mechanistic basis for rational polymer selection and optimization in high drug loaded, and therefore reduced overall volume, ASD formulations.