The strong coupling between hyperelasticity and three-dimensional (3D) large deformations severely complicates the accurate nonlinear mechanical modeling of thin-walled soft pneumatic actuators. This paper presents the systematic design and analytical modeling of a multi-segment pleated soft actuator. While screw theory is introduced to map the spatial deformation of the thin-walled segments, the core contribution lies in rigorously deriving a nonlinear analytical bending model. Specifically, the non-ideal 3D radial expansion inherent to large hyperelastic deformations is integrated as a geometric compensation factor to quantify its coupling effect on macroscopic bending. The proposed theoretical framework is comprehensively verified through multi-source cross-validation involving finite element analysis and physical prototypes. Results demonstrate that the corrected model accurately captures high-pressure strain-stiffening and nonlinear expansion behaviors. The absolute relative error for macroscopic bending angle prediction is constrained within 11%, significantly outperforming traditional uncorrected ideal models. This study lays a rigorous theoretical foundation for the high-fidelity structural design and performance optimization of thin-walled soft structures.

The re-emergence of Monkeypox virus (MPXV) underscores the critical imperative for rapid and deployable diagnostic methods to support effective surveillance and outbreak response. In this study, two recombinase polymerase amplification (RPA)-based assays were developed for MPXV detection by integrating RPA with fluorescence monitoring or lateral flow biosensor (LFB) readouts, referred to as the fluorescent RPA-MPXV and LFB-RPA-MPXV assays, respectively. Both assays were performed under isothermal conditions at 39 °C and efficiently executed within 20 min. The fluorescent RPA-MPXV assay enabled detection via real-time fluorescence monitoring or blue-light-assisted end-point visualization, whereas the LFB-RPA-MPXV assay allowed for instrument-free detection using immunochromatographic strips. Analytical evaluation demonstrated exceptional sensitivity, with a limit of detection of 14 copies using plasmid templates, and uncompromising specificity, as no cross-reactivity was observed against 28 non-MPXV pathogens. Diagnostic applicability was further assessed using simulated clinical specimens, including skin swabs, sputum, and plasma spiked with pseudotyped MPXV. Consistent detection was achieved at concentrations as low as 45 copies across all tested matrices, and all negative controls remained negative, indicating negligible matrix interference and highly stable assay performance. Overall, the fluorescent RPA-MPXV and LFB-RPA-MPXV assays provide rapid and versatile diagnostic tools that complement existing molecular detection methods and support point-of-care testing and field-oriented MPXV surveillance, particularly in settings with limited laboratory infrastructure.

Neonatal sepsis remains a leading cause of infant mortality, yet mechanisms driving concurrent hyperinflammation and immunosuppression remain unclear. Here, we perform single-cell RNA sequencing on 26 blood samples from 18 neonates, spanning acute sepsis, convalescence, and healthy controls. We identify 57 cell subtypes, revealing acute lymphoid depletion and myeloid expansion. S100A8+ myeloid-derived suppressor cell-like (MDSC-like) cells represent a putative cytokine-storm source, potentially amplified by a feedforward S100-TLR4-MYD88 circuit. Innate-like lymphocytes fail to expand, succumbing to apoptosis and exhaustion despite heightened cytotoxicity. CD4+ T cells display mitochondrial dysfunction, while regulatory T cells acquire a hyper-suppressive phenotype via the LGALS9-HAVCR2 axis. CD8+ T cells undergo interferon-driven, innate-like reprogramming before lapsing into exhaustion, and B cells shift toward stress-adaptive, tolerogenic states. Together, our atlas defines a dual pathology in which MDSC-like cell-driven cytokine storm coexists with multi-lineage immunoparalysis, nominating the S100-TLR4 axis and mitochondrial dysregulation as potential therapeutic targets.

Monkeypox virus (MPXV) continues to present a significant global public health challenge, highlighting the critical necessity for rapid molecular diagnostics capable of simultaneously identifying viral infection and discriminating between circulating clades. In this study, we developed a fluorescent and lateral flow biosensor (LFB)-based duplex recombinase polymerase amplification (RPA) platform for the concurrent detection of clade I and clade II MPXV. By integrating the RPA technique with either fluorescence-based real-time monitoring or LFB-based visual readout, the dual-target amplification process is translated into flexible and informative detection outputs. Under single-target conditions, the assays consistently detected clade I and clade II MPXV at concentrations as low as 6.8 & times; 10-1 and 7.7 & times; 10-1 copies/mu L, respectively. Although sensitivity decreased under duplex conditions, both targets remained reliably detectable within a single reaction. The assays further demonstrated high analytical specificity, exhibiting no detectable cross-reactivity with non-MPXV pathogens. The platform demonstrated consistent qualitative performance in simulated clinical samples, including serum, skin swabs, and nasopharyngeal swabs, while indicating potential matrix effects on detection sensitivity. The entire detection workflow, spanning from amplification to result interpretation, can be completed within 25 min. The provision of two selectable readout formats facilitates adaptation to both centralized laboratory analyses and decentralized or point-of-care testing scenarios. Overall, this duplex RPA platform provides a rapid, sensitive, and versatile diagnostic strategy for MPXV surveillance and differential diagnosis.

Inactivated vaccines are critical to COVID-19 immunization, yet the cellular and molecular mechanisms driving pediatric responses warrant detailed characterization. We use single-cell transcriptomics to delineate the immune landscape of peripheral blood mononuclear cells in children vaccinated with CoronaVac. We uncovered a two-phase program: the primary dose establishes a memory foundation via expansion of survival-programmed memory B cells and a shift in CD8+ T cells toward stress adaptation. The booster triggers an effector response marked by plasma cell differentiation, class-switching, and a pan-B-cell type I interferon signature. Concurrently, vaccination reprograms monocytes into a pro-inflammatory, antigen-presenting state enriched for interferon-stimulated genes. This activation is counterbalanced by expanded myeloid-derived suppressor cells and FOXP3+ regulatory T cells, which employ PGE2 signaling to restrain excessive cytotoxicity and orchestrate helper T-cell differentiation. Together, our atlas demonstrates that pediatric immunity to inactivated vaccines is tightly orchestrated, balancing antiviral programs with regulatory mechanisms to ensure safe protection.

Objectives Mycoplasma pneumoniae (MP) is a leading cause of community-acquired pneumonia. Its epidemiology has shifted during and after the COVID-19 pandemic. Current evidence remains predominantly pediatric, creating a significant gap in systematic, all-age data—particularly among adults and hospitalized patients—which limits the development of tailored management strategies. This study employed a multicenter retrospective design to investigate hospitalized cases of MP infection across all age groups in the Hangzhou region of China. Methods Data were retrospectively obtained from inpatients at seven hospitals and four community health centers in Hangzhou, China. Patients of all age groups with acute respiratory tract infection and laboratory-confirmed MP infection were included. Epidemiological trends, age-specific incidence, co-infection pathogen spectra, and antimicrobial resistance-associated mutations patterns were analyzed. Results MP infection presented a “double-peak” pattern synchronous with acute respiratory infections, accounting for 5.2% of 99,730 included patients. Incidence remained < 5% in 2020–2021, increased rapidly, peaked at 12.3% in May 2024, and return to low levels. School-aged children most affected (48.6%); infections in older adults increased by five-fold. Most cases were single infections (83.5%), while 16.5% were co-infections, most commonly with adenovirus and Chlamydia pneumoniae, and the co-occurrence rate between parainfluenza virus and Chlamydia pneumoniae was as high as 73.8%. Macrolide resistance increased significantly, reaching 64.2% in 2023, primarily due to A2063G and A2064G mutations in the 23S rRNA gene. Conclusion The high burden in children, rising incidence in older adults, significant macrolide resistance, and distinct co-infection patterns highlight the need for enhanced, integrated surveillance and tailored stewardship strategies to manage MP infections.

ABSTRACT Escherichia coli (E. coli) is a predominant pathogen causing neonatal sepsis, and rapid pathogen identification is critical for timely antimicrobial therapy. However, conventional diagnostic methods are often limited by prolonged turnaround time, or insufficient sensitivity. In this study, we developed a multiple cross-displacement amplification assay coupled with a lateral flow biosensor (MCDA-LFB) targeting the uidA gene for rapid visual detection of E. coli. Using a set of 10 specific primers, the reaction conditions were optimized at 63°C for 40 min. The analytical limit of detection was 50 fg genomic DNA per reaction, and no cross-reactivity was observed among the tested non–E. coli strains. Clinical performance was evaluated using 75 residual sterile body fluid specimens, including 58 blood and 17 cerebrospinal fluid samples, collected from neonates who were clinically diagnosed with sepsis or bacterial meningitis, and in whom E. coli infection was considered highly likely on the basis of the overall clinical assessment. Results were compared with quantitative PCR (qPCR) and conventional culture. In this cohort, the MCDA-LFB assay showed complete agreement with qPCR (κ = 1.0) and detected 56.0% (42/75) positive samples, compared with 14.7% (11/75) by culture (P < 0.001). The entire workflow, including nucleic acid extraction, amplification, and visual readout, was completed within approximately 1 h. These findings suggest that the E. coli-MCDA-LFB assay provides a rapid, sensitive, and cost-effective approach for E. coli detection and may be useful for early diagnosis in clinical neonatal sepsis, particularly in point-of-care and resource-limited settings.IMPORTANCENeonatal sepsis requires rapid and accurate pathogen identification to support timely antimicrobial therapy, yet conventional blood culture is often slow and has limited sensitivity, particularly in neonates with small sample volume or prior antibiotic exposure. In this study, we developed a visual multiple cross displacement amplification–lateral flow biosensor (MCDA-LFB) assay for rapid detection of Escherichia coli, a leading cause of neonatal sepsis. The assay enables visual readout within approximately 1 h, shows femtogram-level analytical sensitivity, and demonstrates complete agreement with quantitative PCR (qPCR) in our clinical validation cohort. Owing to its operational simplicity and minimal equipment requirements, this method may serve as a useful approach for rapid detection of neonatal E. coli infection, particularly in resource-limited clinical microbiology settings.

Salmonella enterica can persist in low-moisture foods and shows enhanced dry-heat resistance under low water activity, posing significant food safety challenges. However, the genetic basis of extreme dry-heat resistance and its relationship with other processing stresses remain unclear. In this study, twelve S. enterica strains were screened for dry-heat treatment at 60 °C and 80 °C, with S. Infantis CICC21649 identified as the most resistant strain. Comparative genomics and transcriptional analysis identified candidate genes related to envelope integrity and regulation, including gtrB and dam. Deletion of the chromosomal gtrB homolog reduced dry-heat resistance, producing an additional 0.91-log10 reduction relative to the parent strain at 80 °C. Deletion of dam caused broader stress sensitivity, reducing resistance to both dry heat and high hydrostatic pressure, with the stronger phenotype observed under high hydrostatic pressure. Proteomic analysis of the chromosomal gtrB homolog mutant revealed broad alterations in envelope-associated proteins, transport functions, oxidative stress pathways, and central metabolism under dry-heat stress. These findings indicate that the chromosomal gtrB homolog is an important contributor to extreme dry-heat resistance, whereas dam contributes to resistance against both dry-heat and high hydrostatic pressure, likely through a broader regulatory role in stress adaptation. These results reveal distinct structural and regulatory layers underlying stress adaptation in S. enterica and provide practical guidance for low-moisture food processing by highlighting the need to account for strain-dependent and stress-specific resistance during process validation.

RATIONALE:The host immune determinants that distinguish protective from life-threatening responses to influenza are poorly understood. Identifying drivers of immunopathology in the human lung is critical for developing potential therapies. OBJECTIVES:To define the cellular and molecular immune landscape of the lung in mild vs severe influenza and to identify key cellular states and pathways associated with disease severity. METHODS:We generated a large-scale single-cell atlas by sequencing more than 520 000 cells from the bronchoalveolar lavage fluid of 88 nonimmunocompromised adult individuals with mild or severe influenza A and healthy controls. Key findings were validated by flow cytometry and protein quantification, and machine-learning models were used to identify predictive signatures. MAIN RESULTS:Severe influenza was characterized by profound pulmonary lymphopenia and a massive influx of functionally dysregulated neutrophils. The infiltrating neutrophils were primed for extracellular trap formation, driving a cytokine storm via the S100A8/A9/A12-TLR4 and CXCL8-CXCR1/2 axes. This pathology coincided with the depletion and functional impairment of resident alveolar macrophages and an expansion of pro-inflammatory, monocyte-derived macrophages that amplified neutrophil recruitment. Lymphopenia in severe disease arose from synergistic cell-death programs, while remaining lymphocytes exhibited a dysfunctional state of concurrent exhaustion and hypercytotoxicity. Mild influenza featured a coordinated adaptive immune response, distinguished by an enrichment of T follicular helper cells and plasma cells. Machine-learning models identified robust cellular and transcriptional signatures predictive of disease severity. CONCLUSIONS:Our atlas defines the divergent immune trajectories in influenza, revealing specific cellular states and pathways that drive immunopathology and provide novel targets for host-directed therapies.

Soft actuators, with their compliance, adaptability, and lightweight characteristics, have become a key research focus driving the development of soft robotics. This paper provides a systematic review of recent advances in pneumatic soft actuators, with emphasis on actuation design, fabrication techniques, and pose measurement methods. Common structural types are categorized and summarized, and the latest design strategies involving different materials and motion modes are introduced, together with an analysis of how structural variations influence actuator performance. Current fabrication methods are further examined, highlighting both the advantages and limitations of molding, 3D printing, and other techniques in the development of complex, high-performance actuators. Various measurement approaches are systematically reviewed, including contact-based and non-contact methods. Overall, the design, fabrication, and measurement of pneumatic soft actuators are closely related to their performance, controllability, and application potential, making them of significant importance for research in this field. This review establishes an accessible reference framework, summarizes key challenges, and outlines future directions such as intelligent design, multifunctional system integration, and cross-disciplinary applications, thereby providing a comprehensive overview of the state of the art and fostering future innovation.

Human adenovirus (HAdV) is a leading cause of acute respiratory tract infections (ARTIs) in children. The high prevalence of HAdV serotypes 3 and 7 in regions such as China presents a significant public health challenge. Here, we propose a one-pot assay that integrates multiple cross displacement amplification (MCDA) with CRISPR-Cas12b for the detection of HAdV-3 and HAdV-7, termed HAdV-MCDA-One. In this system, MCDA provides exponential target amplification, while the collateral cleavage activity of Cas12b enables secondary signal amplification. The entire reaction is performed isothermally at 60°C in a single tube, providing a fluorescent readout within 50 min, making the assay suitable for point-of-care testing (POCT). Leveraging the single-base recognition capability of CRISPR-Cas12b, the assay demonstrates high specificity, with no cross-reactivity observed against the other 13 identified pathogens. The limit of detection was determined to be 1.59 copies per reaction using target plasmids. Moreover, when evaluated with 96 clinical pharyngeal swabs, the assay showed 100% concordance with quantitative PCR (qPCR), confirming its clinical reliability. These results demonstrate HAdV-MCDA-One as a rapid and robust tool for HAdV-3 and HAdV-7 detection, with significant potential for clinical diagnosis and public health surveillance.

Influenza A virus (IAV) infection has a wide clinical spectrum, from mild illness to life-threatening pneumonia, yet the underlying immune determinants of disease remain poorly defined. Here, we generated a large-scale single-cell transcriptomic atlas from peripheral blood, profiling more than 612,010 cells from 97 individuals, including healthy controls, and patients with mild, severe, or convalescent IAV infection. Our findings uncovered a core immune dichotomy that determines clinical severity: a protective, monocyte-centric antiviral state in mild disease versus a pathological, neutrophil- and myeloid-derived suppressor cell (MDSC)-driven hyperinflammatory state in severe infection. Severe disease was marked by a peripheral hyperinflammatory state, driven by specific monocyte and neutrophil subsets via the S100A8/9/12-TLR4/RAGE signaling axis, and was coupled with the expansion of granulocytic MDSCs that likely contribute to T cell paralysis. In contrast, mild disease was associated with a protective, monocyte-centric response characterized by robust antiviral interferon signaling and enhanced antigen presentation. This functional divergence extends to the adaptive immune system, where mild disease was associated with CD8+ T cells displaying a balance of high cytotoxicity and regulated exhaustion. In severe illness, however, T cells become profoundly dysfunctional, exhibiting signatures of metabolic stress and apoptosis alongside the emergence of pathogenic, pro-inflammatory regulatory T cells. Together, our atlas provides a high-resolution immunological blueprint of human IAV infection, delineates the cellular states and pathways that govern clinical trajectories and offers a critical resource for developing host-directed therapies.

Background:The prospective association between C-reactive protein (CRP) and gestational diabetes mellitus (GDM) is inconsistent, and the independent association of tumor necrosis factor-α (TNF-α) with GDM risk remains unclear. We aimed to prospectively evaluate associations of early-pregnancy CRP and TNF-α with incident GDM and updated the evidence through a meta-analysis. Methods:Serum CRP and TNF-α levels were measured at 6-15 weeks of gestation among 332 GDM and 664 matched controls. Conditional logistic regression was used to compute odds ratios (ORs) and 95% confidence intervals (CIs). PubMed and EMBASE were searched up to June 18, 2025, to identify prospective studies that investigated associations of CRP and TNF-α levels with GDM. Results:After multivariable adjustment, OR for GDM was 1.75 (95% CI 1.07, 2.84) for the extreme-quartile comparison of CRP and a non-linear association was noted (P-nonlinear = 0.03). TNF-α was associated with a 1.82-fold (95% CI 1.07, 3.09; P-trend = 0.04) higher risk of GDM. CRP and TNF-α levels were positively associated with the risk of GDM, independent of each other and of pre-pregnancy body mass index (BMI). The meta-analysis confirmed the positive association between CRP and GDM risk, independent of BMI. CRP levels were associated with adverse metabolic traits (e.g. fasting insulin, homeostasis model assessment for insulin resistance [HOMA-IR], and HOMA-β), while TNF-α was not. Conclusion:Elevated CRP and TNF-α levels in early pregnancy were independently associated with increased risks of GDM. While our findings support potential roles of inflammation in GDM, specific pathways involved should be examined to identify novel targets for the prevention of GDM.

OBJECTIVE: This study analyzed the frequency and patterns of pediatric neurological disorders and investigated changes in the inpatient disease spectrum following the COVID-19 pandemic. METHODS: A retrospective analysis was conducted on 2,981 children admitted to the Department of Neurology at the Seventh Medical Center of PLA General Hospital in Beijing, China, between January 2019 and December 2024. RESULTS: According to the ICD-10 criteria, the most common diagnoses were “nervous system diseases” (39.74%) and “mental and behavioral disorders” (31.98%). The pandemic led to a significant increase in the proportion of inpatients with mental and behavioral disorders, whereas the proportion of hospitalizations for nervous system diseases among all pediatric neurology admissions decreased. Furthermore, an increase in the number of autism spectrum disorder (ASD) inpatients was observed starting in 2022, with a more notable increase from 2023 to 2024. CONCLUSION: This study provides detailed insights into shifts in the inpatient disease spectrum of pediatric neurology from 2019 to 2024. These findings, which are based on hospitalized patients, highlight changes in healthcare utilization patterns and may inform hospital resource planning. These findings provide a foundation for future research that incorporates outpatient data to understand the pandemic’s impact on pediatric neurological disorders.

The seamless and efficient integration of multiple detection modes─such as electrochemical, colorimetric, magnetic, thermal, and fluorescent signaling─into a single sensing platform remains a significant challenge in advanced sensing technology. In this work, we developed a trimodal biosensor based on homogeneous polydopamine (PDA)/Fe3O4/CP (CP: capture probe) nanocomposites and three-dimensional hybridization chain reaction-templated silver nanoclusters (3D-HCR/AgNCs) for the ultrasensitive and precise detection of nucleic acids. Briefly, the presence of the target KRAS gene segment initiates a hybridization chain reaction (HCR), which acts as a bridge linking capture probes (PDA/Fe3O4/CP) and signal probes (3D-HCR/AgNCs). The HCR bridge hybridizes with both components, forming a PDA/Fe3O4/CP/HCR-bridge/3D-HCR/AgNCs complex that can be magnetically separated. Since DNA-templated AgNCs generate measurable signals across three distinct modes─electrochemistry, fluorescence, and visible/UV-vis spectroscopy─a fully integrated trimodal sensing platform is realized. The results from these three modes can be cross-verified, significantly enhancing the reliability of the assay. Leveraging homogeneous PDA/Fe3O4/CP nanocomposites and a strategically designed blocker, this strategy enables straightforward, one-pot detection of target nucleic acids without the need for vibration, electrode modification, or multiple washing and separation steps. Importantly, beyond serving as a signal amplifier, the 3D-HCR structure also promotes aggregation-induced emission enhancement of AgNCs, thereby increasing the sensitivity of the fluorescence mode. This work introduces a facile and efficient approach that paves the way for the construction of integrated trimodal sensing platforms.

Rice blast, caused by Magnaporthe oryzae (M. oryzae), is a severe threat to rice production globally. The pathogen counters rice immunity by secreting effectors that disrupt host defenses. In this study, we conducted a comprehensive genome-wide screening to identify candidate secreted effector proteins (CSEPs) in M. oryzae. Using a new bioinformatics pipeline, we predicted 577 CSEPs and analyzed their sequence features and functional annotations. We found that these effectors have distinct sequence signatures, such as high cysteine content, and are involved in infection and immune suppression. Phylogenetic analysis revealed M. oryzae's close relationship with other pathogenic fungi and the conservation of certain CSEPs across species. Expression analysis during infection indicated a role of CSEPs in the pathogenic process and the ability to inhibit plant necrosis. Finally, we validated the function of three candidate effector proteins through gene disruption mutant analysis including pathogenesis testing in rice. This study provides a foundation for understanding M. oryzae pathogenicity and may aid in developing resistance strategies against rice blast.

The aim of this study was to develop a method based on multiple cross displacement amplification (MCDA) and real-time fluorescence technique for rapid, highly sensitive and specific detection of Streptococcus pyogenes (Group A Streptococcus, GAS). A set of 10 primers targeting the speB gene of GAS was designed for the MCDA reaction. According to the principle of real-time MCDA detection, the core primer was further modified with a restriction endonuclease recognition sequence, a fluorophore, and a quencher. The optimal reaction temperature for the assay was determined based on the performance of the MCDA amplification products. The detection limit was evaluated using tenfold serial dilutions of GAS genomic DNA templates. To assess specificity, the assay was tested using genomic DNA from 3 GAS strains and 29 non-GAS strains. To evaluate clinical application of the GAS real-time MCDA assay, 56 clinical samples were analyzed and compared with the lateral flow biosensors (LFB) method and PCR. The GAS real-time MCDA method was performed using a fluorescence instrument at 63℃ for 40 min. The method demonstrated high sensitivity, with a detection limit of 50 fg, and showed no cross-reactivity with other pathogens. The GAS real-time MCDA assay described here offers a new and valuable diagnostic tool for the reliable and rapid detection of GAS.

Acute respiratory infections (ARIs) in children present complex interactions between respiratory pathogens and host, contributing to high morbidity, hospitalization, and mortality. In this cross-sectional study, 2,044 pediatric in-patient samples were tested for 198 pathogens using targeted next-generation sequencing (tNGS). Pathogen spectrum and co-infection pattern were analyzed alongside blood-test results and clinical data. Pediatric ARIs presented staggered viral epidemics and age-specific infection patterns from 2022 to 2023. Viral infections were predominant in children under 3 years (67.5%). Specific viral-bacterial co-infections were associated with disease severity, including human metapneumovirus (HMPV)-Streptococcus pneumoniae (OR = 3.172, 95% CI: 1.257-8.009), human parainfluenza virus (HPIV)-Fusobacterium nucleatum (3.016, 1.051-8.653), and HPIV-S. pneumoniae (2.825, 1.007-7.927). Subnormal levels of prealbumin and creatinine served as biomarkers for homeostasis disruption in viral-bacterial co-infections. From June to December 2023, Mycoplasma pneumoniae (MP) became the dominant pathogen in preschool (31.3%) and school-age (45.3%) children. High MP loads were correlated with elevated eosinophil, IL-4, and IL-6 and decreased IgG and IL-2, linking to airway hyperreactivity and inflammation. At low MP loads, patients co-detected with human herpesvirus (HHV) (50.0%), or Haemophilus influenzae (40.0%) had a high proportion of severe cases. This study delineated the complex pathogen landscape of pediatric ARIs and highlighted the major role of viral-bacterial and MP-bacterial co-infections. Monitoring these pathogens through tNGS can aid in precise diagnosis and targeted treatment to improve clinical outcomes in children.IMPORTANCEMoving beyond the outdated "one germ, one disease" model, this study characterizes the etiological and epidemiological landscape and highlights co-infection patterns of virus-bacterium or MP-bacterium in 2044 pediatric respiratory in-patient children across three epidemic phases. Specific pathogen combinations are associated with clinical severity, as well as host immune and metabolic profiles. Our findings underscore the necessity of detecting pathogen-pathogen and host-pathogen dynamics, rather than individual pathogens, thereby informing precision diagnostics and targeted intervention strategies.