Biology subjects

Guo, Y.

Publications and source records attributed to Guo, Y..

At least 37 records · Page 2Linked to original sources

bioRxiv2026

Acute caffeine ingestion differentially alters motor unit behaviour during submaximal and fatiguing contractions

Background: Caffeine is widely used as an ergogenic aid, yet its effect on neuromuscular function remains heterogeneous across exercise conditions. Although caffeine is known to enhance central nervous system excitability through adenosine receptor antagonism, the extent to which these effects modify spinal motor output remains unclear. Therefore, the present study aimed to characterise acute caffeine-induced alterations in neuromuscular properties during submaximal and sustained fatiguing contractions of the knee extensors. Methods: Seventeen healthy adults completed a randomised, placebo-controlled crossover study (3 mg/kg caffeine). Circulating serum caffeine concentrations were periodically quantified, while motor unit (MU) discharge behaviour, and delta F were derived from high-density surface electromyography. Statistical significance was set at p = 0.05. Results: No intervention effects were observed for MVC, force steadiness, or time to task failure (all p > 0.05). During graded contractions, vastus lateralis MU discharge rate was higher under caffeine (p < 0.001), and discharge variability was lower (p = 0.016). Recruitment and derecruitment thresholds were significantly greater under caffeine at 50% MVC (both p < 0.012), whereas paired MU analyses of delta F was unchanged (p = 0.083). During sustained contractions, caffeine altered fatigue-related trajectories of both MU discharge and discharge variability (intervention * phase interaction, p < 0.011). Conclusions: Acute caffeine ingestion alters motor unit behaviour without enhancing maximal force or fatigue resistance. These effects vary with contraction intensity and fatigue state but are not accompanied by a detectable change in discharge rate hysteresis in the vastus lateralis.

physiology↗

bioRxiv2026

Discriminating rare disease cases from their controls based on observed and excluded phenotypes

Rare diseases are individually uncommon but collectively prevalent. Their primary clinical challenge lies not in treatment but in diagnosis. In the early stages of clinical management, it is frequently unclear whether the observed phenotypes are associated with a rare disease. Leveraging machine learning methods to mine latent associations between these phenotypes and rare diseases for early diagnosis offers a viable strategy to alleviate this diagnostic dilemma. In the present study, we demonstrated that machine learning can effectively discriminate rare disease cases from their controls using observed and excluded phenotypes as features. Among them, the Random Forest model achieved the best classification performance with a certain degree of generalizability. Based on further analysis of the feature selection results, we conclude that the two factors, specificity and occurrence count, are important for phenotype selection in rare disease discrimination, and comparable importance should be attached to both observed and excluded phenotypes, during feature construction.

bioinformatics↗

bioRxiv2026

Antibody evasion and receptor binding of SARS-CoV-2 RW.1.1

The continued evolution of SARS-CoV-2 is shaped by changes in antibody evasion and receptor engagement that influence viral fitness. RW.1.1, an emerging descendant of XFJ carrying five additional spike substitutions, has recently increased in frequency in North America. Here, we characterized the serum antibody evasion, monoclonal antibody sensitivity, and ACE2 receptor engagement of RW.1.1. Pseudovirus neutralization assays using sera from adults showed that RW.1.1 was not more resistant to serum neutralization than currently circulating variants, with neutralizing titers comparable to XFG, BA.3.2.2, and NB.1.8.1 in the tested cohort. Neutralization of RW.1.1 also did not differ significantly among adults, children, and infants and toddlers. Despite the absence of increased overall serum antibody resistance, monoclonal antibody neutralization assays revealed substantial resistance to several RBD class 1 antibodies and increased resistance to a subset of class 1/4 antibodies. Moreover, RW.1.1 exhibited reduced ACE2 receptor engagement compared with XFG, which itself has reduced receptor engagement relative to earlier JN.1 subvariants. Thus, RW.1.1 has continued to expand despite a further reduction in receptor engagement and without a substantial increase in overall serum antibody evasion. These findings suggest that the fitness of emerging SARS-CoV-2 variants may depend not only on the magnitude of antibody evasion but also on the specific components of the polyclonal antibody response that are evaded, highlighting the increasingly complex interplay between population immunity and receptor engagement in shaping SARS-CoV-2 evolution.

microbiology↗

bioRxiv2026

Cryo-EM structure of detergent-bound Vibrio cholerae MakA captures a candidate intermediate between the soluble form and mature assembly of an α-pore-forming toxin

The tripartite toxin MakA/MakB/MakE from Vibrio cholerae belongs to a family of -pore-forming toxins that undergo extensive conformational rearrangements upon interacting with membrane. While soluble structures of MakA, MakB and MakE have been determined, the structural basis of membrane association and pore assembly remains poorly understood. Here, we report a cryo-electron microscopy structure of a detergent-associated form of MakA at a global resolution of 3.13 Angstrom, with focused refinement of a flexible region improving the local resolution to 2.68 Angstrom. This structure consists of a trimer of MakA asymmetric homodimers, in which each subunit adopts a transmembrane -helical conformation distinct from the previously reported soluble structure. The asymmetric homodimer closely resembles the building block observed in the previously reported helical MakA assembly and is structurally similar to the homodimeric building blocks of the Aeromonas hydrophila AhlB and Serratia marcescens SmhA pores, and the heterodimeric building blocks of the Xenorhabdus nematophila XaxAB and Yersinia enterocolitica YaxAB pores, suggesting a conserved assembly principle among distantly related -pore-forming toxins. Complementary hydropathy analysis, AlphaFold2 modeling, and liposome-based assays reveal distinct roles for the three components, with MakA acting as the primary membrane-inserting subunit, MakE exhibiting low but consistent membrane-association, and MakB alone remaining largely soluble. Together, these results support a model in which the MakA asymmetric homodimer represents a plausible early membrane-insertion unit and building block for pore assembly, while the trimer of dimers represents an early assembly intermediate.

microbiology↗

bioRxiv2026

Systemic hypoxia drives glycogen-fueled progression of lung adenocarcinoma

In advanced stages, lung adenocarcinoma obstructs airways and disrupts ventilation-perfusion relationships in the lung, causing systemic hypoxemia and enabling a feed-forward loop that accelerates malignancy. Systemic hypoxemia is also experienced due to common respiratory comorbidities such as chronic obstructive pulmonary disease (COPD) and obstructive sleep apnea (OSA), potentially accelerating malignancy. In a statewide electronic health record network, pre-existing COPD (598 matched pairs) or sleep apnea (235 matched pairs) independently predicted worse survival following incident lung cancer diagnosis. Since the mechanistic basis of the link between malignancy and hypoxia is not well understood, we created systemic hypoxia in KrasLSL-G12D/+;Trp53fl/fl (KP) mice by delivering low inspired oxygen concentrations (8% inspired oxygen; 8 h daily). Hypoxia nearly doubled tumor multiplicity and selectively remodeled cancer central carbon metabolism. Spatially resolved metabolomics revealed marked tumor-compartment glycogen accumulation, elevated tricarboxylic-acid cycle intermediates, and depleted glycolytic pools. Quantitative proteomics across cellular models and autochthonous tumors demonstrated that systemic hypoxia drives glycogen mobilization selectively through the lysosomal enzyme acid -glucosidase (GAA). Tumor-cell-autonomous deletion of GAA eliminated the hypoxia-driven growth advantage and disrupted downstream anabolic biosynthetic pathways. Thus, systemic hypoxia drives lung adenocarcinoma expansion by mobilizing lysosomal glycogen reserves through GAA to sustain proliferative growth.

cancer biology↗

bioRxiv2026

In Vivo Screening for a Promising Antiparasitic Agent Against Neobenedenia melleni in Epinephelus fuscoguttatusfemale x E. lanceolatusmale and Identification of Its Potential Target

Monogenean ectoparasites, particularly Neobenedenia species, cause severe economic losses in mariculture. Here, ectoparasites isolated from cultured hybrid groupers (Epinephelus fuscoguttatus[female] x E. lanceolatus[male]) were confirmed as Neobenedenia melleni based on ITS1 phylogeny. In vivo screening of six structurally diverse compounds identified compound D (CAS No. 206111-37-7), a 5,6-dihydropyridine derivative, as the most effective antiparasitic agent, achieving 76.54% efficacy at 0.5 mg/L in a 90 min bath treatment. Dose-response assays demonstrated that 0.7 mg/L compound D achieved 95.23% antiparasitic efficacy without causing evident tissue damage or cytotoxicity to GF-1 cells. Ultrastructural observation by scanning electron microscopy revealed marked tegumental alterations, including deep fissures and extensive surface folding, in treated parasites. Molecular docking against ten candidate proteins identified {beta}-tubulin as the most favorable docking target, with a binding energy of -6.53 kcal/mol and three hydrogen-bond interactions, suggesting that {beta}-tubulin may be involved in the antiparasitic activity of compound D. Overall, these findings highlight compound D as a promising lead candidate for short-bath therapy against N. melleni and suggest that cytoskeletal disruption through {beta}-tubulin interaction represents a plausible mechanism of action.

zoology↗

bioRxiv2026

Computer-aided drug screening of anti-Neobenedenia melleni drugs based on annexin B1 in farmed pearl grouper

Monogenean capsalids of the genus Neobenedenia are widespread parasites of wild and farmed marine fish, and represent a great threat to the mariculture of grouper in China. Fishery drug development to screen and find effective compounds to control and prevent the disease is urgent needed, considering the vast production of grouper in China (294 ktons in 2025). Annexins have been discovered in Neobenedenia and other parasites, and marked differences between the parasite annexins and those of the hosts make them potentially attractive drug targets for anti-parasite therapeutics. Herein, we utilized computer-based drug discovery screens using unique Neobenedenia melleni annexin B1 and a database of 1,456,161 small molecules. The 3D structure of annexin B1 was firstly modeled by three different protein prediction tools, namely AlphaFold 3, SWISS-MODEL, and I-TASSER, of which the most accurate protein structure was used as the drug target for the following structure-based virtual screening. In vivo experimental validation of 11 compounds after molecular docking shows that abamectin (Aba) has the most effective anti-Neobenedenia bioactivity at the concentration of 0.16 mg/L as the initial screening concentration. Given its low toxicity to host grouper (24 LC50=0.254 mg/L), abamectin was chose for further investigation. A 24 h bath exposure successfully lowered the parasitic load in infected grouper, yielding an 24 h EC50 of 0.033 mg. To elucidate the anti-parasite mechanism, long-timescale molecular dynamics simulations (1000 ns) of annexin B1 and Aba was conducted, which allowed for atomic and molecular-level analysis of the essential protein motions involved in the interaction of annexin B1 and its substrate. The interaction profile between annexin B1 and abamectin was dominated by hydrophobic contacts and water bridges, involving residues TYR-210, GLU-214, GLU-244, and SER-247, which path a way for further drug optimization.

zoology↗

bioRxiv2026

An AI-assisted platform for quantitative histopathological analysis in interstitial lung disease

Interstitial lung diseases (ILDs) are heterogeneous pulmonary disorders characterized by chronic inflammation and/or fibrosis. 30-40% of ILD patients develop fibrotic disease that is associated with progressive respiratory decline and poor prognosis, particularly in idiopathic pulmonary fibrosis. Current antifibrotic therapies slow disease progression but do not reverse fibrosis, highlighting the need for improved therapeutic strategies. Robust histopathological evaluation in preclinical models is essential for drug development; however, conventional scoring systems are semi-quantitative, labor-intensive, subject to inter-observer variability, and rely on limited field sampling. Here, we introduce FibroSight, a standalone platform for compartment-resolved quantification of lung remodeling in Sirius Red-stained sections. By integrating deep learning- based structural segmentation with color-based feature extraction, FibroSight enables highly automated whole-lobe analysis without requiring complex computational setup. The platform quantifies complementary remodeling parameters, including parenchymal collagen fraction, parenchymal tissue density, nuclear area fraction, parenchymal airspace fraction, and airway- and vascular-associated remodeling. Validated in the bleomycin-induced fibrosis model, FibroSight-derived metrics strongly correlated with expert Ashcroft scoring and showed stronger associations with histological severity than corresponding outputs from a semi-automated ImageJ-based workflow. The platform further distinguished inflammatory from fibrotic remodeling in influenza-induced lung injury and demonstrated translational proof-of-concept applicability in human ILD biopsy specimens. By enabling scalable, reproducible, and multi-compartment histological quantification, FibroSight provides a practical framework for objective assessment of lung remodeling. This approach expands conventional fibrosis evaluation by integrating fibrotic, inflammatory, airway, and vascular-associated readouts, supporting more precise analysis of disease mechanisms and therapeutic responses in preclinical and translational ILD research.

pathology↗

bioRxiv2026

A modular architecture of human cellular aging across organs and diseases

Aging is a heterogeneous biological process in which different cellular systems undergo molecular remodeling at distinct rates, yet whether human cellular aging follows an organized architecture across organs remains unclear. Here, we integrate a multi-organ human single-cell transcriptomic atlas with plasma proteomic profiles from approximately 50,000 participants to reconstruct cellular aging states at population scale. By projecting cell-type-enriched molecular signatures onto circulating proteins, we characterize aging patterns across 128 organ-cell type pairs and identify 14 cellular aging modules comprising conserved cross-organ programs and organ-specific aging states. These modules reveal cellular identity as a dominant organizing axis of human aging that transcends anatomical boundaries. Module-level aging states uncover substantial inter-individual heterogeneity, with 34% of individuals exhibiting extreme aging deviation in at least one cellular module. Cellular aging modules exhibit distinct temporal trajectories, with structural and tissue-resident modules showing earlier remodeling than immune lineages. The modular organization of cellular aging is reflected in disease susceptibility, with accelerated aging of specific modules, particularly epithelial aging, showing broad associations with disease burden and mortality. Longitudinal analyses further demonstrate the stability and clinical relevance of cellular aging states, whereas lifestyle, metabolic and pharmacological factors show selective relationships with individual aging programs. Together, our study establishes a modular framework for understanding human cellular aging and reveals an organization of biological aging that may help explain individual differences in healthspan.

cell biology↗

bioRxiv2026

Single-cell profiling reveals MMP7-associated epithelial transition and immune-stromal remodeling in esophageal carcinogenesis

The progression from reflux esophagitis (RE) to Barretts esophagus (BE) and esophageal adenocarcinoma (EAC) is a major route of inflammation-associated esophageal tumorigenesis, but the cellular transitions involved remain incompletely understood. We performed single-cell transcriptomic profiling of RE, BE, and EAC tissues and integrated public EAC datasets, yielding 51,283 cells across the RE-BE-EAC continuum. The analysis identified stage-associated epithelial and microenvironmental differences. Pseudotime analysis placed Basal epithelial cells along two differentiation branches, one toward mature epithelial states and the other through proliferative and gastric-type metaplastic states toward copy number alteration-bearing malignant epithelial cells. MMP7 increased along this malignant branch, was enriched in cancer-associated epithelial cells. Its knockdown reduced the proliferation and migration of OE19 and OE33 cells, supporting an association with proliferative and migratory phenotypes. In parallel, the microenvironment shifted toward immune suppression and stromal remodeling, with increased T-cell exhaustion, macrophage polarization toward tumor-associated states, and enrichment of CAF-like myofibroblasts. Ligand-receptor analysis further suggested an epithelial-centered communication network involving MDK-SDC2/LRP1/NCL interactions between malignant epithelial cells and fibroblast, T-cell, and myeloid compartments. These results identify MMP7-associated epithelial states and candidate MDK-related communication pathways for further longitudinal and functional validation.

cancer biology↗

bioRxiv2026

Hologenomics of xylotrophic bivalves reveals a minimalist, remote-acting evolutionary strategy of wood digestion

Wood constitutes the largest reservoir of biogenic carbon on Earth, yet remarkably few animals can exploit it. While terrestrial wood-feeders like termites rely on highly diverse gut microbiomes, xylotrophic marine bivalves have evolved a fundamentally different approach: a spatially segregated system where intracellular gill symbionts produce enzymes that act remotely within a nearly sterile cecum. However, the genetic and evolutionary basis of this unique symbiosis remains largely elusive. Here, we integrate hologenomics, transcriptomics, and biochemistry of a shallow-water shipworm (Teredo navalis) and a deep-sea borer (Xyloredo sp.). We find that despite diverging approximately 147 million years ago and occupying drastically different habitats, these bivalves maintain a strictly conserved ancestral karyotype and a shared genomic architecture for wood digestion. Our models reveal a clear host-symbiont division of labor. The host genome is specialized for lignin modification and targeted enzyme transport, whereas a highly streamlined symbiont community is responsible for core polysaccharide degradation. Central to this minimalist strategy is a lineage-specific GH5-GH6 dual-catalytic enzyme. By sharing amino acids across proximal binding pockets, this fusion protein unites endo- and exo-cellulase activities, enabling highly synergistic cellulose cleavage without the need for complex microbial communities. Ultimately, our comparative analysis with terrestrial models demonstrates that these marine invertebrates achieve efficient biomass degradation not through microbial expansion, but through extreme functional streamlining and molecular innovation, offering a distinct evolutionary paradigm for marine carbon cycling.

evolutionary biology↗

bioRxiv2026

Effect of nuclear electrostatic potential on the nucleoplasmic distribution of histones and nucleosome stability

Differences in the density of heterochromatic and euchromatic regions are often assumed to play an important role in determining the transcriptional state of chromatin by influencing its accessibility to transcription factors. Yet, experiments show that even the most densely packed chromatin domains are readily accessible to transcription factors. Thus, the molecular mechanisms underlying differences in the transcriptional states of hetero- and euchromatin remain poorly understood. In this study, using electrically charged mEGFP probes, we demonstrated that heterochromatin and euchromatin differ not only in density but also in the magnitude of the local electrostatic potential. Furthermore, the nucleoplasmic distribution of electrically charged proteins, such as histone-chaperone complexes, was found to correlate with the local electrostatic potential. Estimates based on experimental data have also shown that the difference in electrostatic potentials between heterochromatic and euchromatic regions could lead to unequal nucleosome stability in them, which was successfully confirmed experimentally. Subsequent theoretical calculations showed that this could shift the balance in the DNA-binding competition between transcription factors and nucleosomes, thereby explaining experimental observations that heterochromatin is less transcriptionally active than euchromatin. Overall, our study suggests the existence of a nuclear electrostatic potential-mediated pathway that may be involved in the regulation of gene transcription.

biophysics↗

bioRxiv2026

pHaseMD4AI: Phase-Space Dynamics Dataset with Chemical and pH Perturbations for Physically and Kinetically Consistent Biomolecular AI

Protein function emerges from dynamic conformational ensembles and transitions that are challenging to characterize experimentally and computationally. Recent advances in generative AI have created new opportunities for learning molecular thermodynamics, kinetics, and conformational evolution directly from simulation data, but progress is limited by the availability of large-scale datasets that combine rigorous sampling, complete phase-space information, and diverse physicochemical perturbations. Here, we present pHaseMD4AI, a molecular dynamics dataset that combines a globally equilibrated peptide branch with a protein-scale constant-pH molecular dynamics (CpHMD) branch spanning hundreds of soluble proteins. The peptide branch includes a complete set of canonical tripeptide and tetrapeptide systems together with post-translationally modified (PTM) and protonation-state datasets, providing synchronized atomic coordinates (R), velocities (V), forces (F), and Markov state model-based kinetic annotations. An accompanying web portal (https://isb.zju.edu.cn/md4ai/) enables users to browse, visualize, and download trajectories, annotations, and metadata. As an example application, we demonstrate a sequence-based model that can predict residue-level equilibrium dihedral distributions from sequence. pHaseMD4AI provides a resource for developing and benchmarking molecular machine learning methods while supporting broader studies of biomolecular dynamics under sequence, post-translational modification, and protonation-state perturbations.

bioinformatics↗

bioRxiv2026

Thiooxazole Formation on a Nontypeable Haemophilus influenzae Virulence Factor Requires a Mixed-Valent Diiron Cofactor

The multinuclear nonheme iron-dependent oxidative enzyme (MNIO) family employs a multi-iron cofactor to catalyze a range of post-translational modifications (PTMs) in the biosynthesis of ribosomally synthesized, post-translationally modified peptide (RiPP) natural products. While significant progress has been made toward understanding the range of chemical transformations performed by MNIOs, the nature of the iron cofactor has only been investigated in one instance. Here, we examine the MNIO involved in oxazolin biosynthesis to gain further insight into the metallocofactors employed by this impressive family of enzymes. Oxazolin, a RiPP virulence factor from nontypeable Haemophilus influenzae, contains six copper-binding 5-thiooxazole groups installed by the MNIO HvfB. Weak interactions between HvfB and its required partner protein, HvfC, motivated genetic fusion of the two proteins, which yielded an effective mimic of the protein complex with high enzymatic activity. While HvfB binds up to three iron ions, concerted EPR, ENDOR, and Mossbauer spectroscopic characterization of the active protein reveals that accumulation of a mixed-valent diiron(II/III) cluster correlates with 5-thiooxazole product formation. This oxidation state is attained only in the presence of HvfC, revealing a new role for the partner protein in modulating the iron cofactor. Site-directed mutagenesis of metal-coordinating residues was used to probe the function of the third iron-binding site. This work clarifies the nature of the active iron cofactor for oxazolin maturation, providing a second example of a mixed-valent diiron oxidase in RiPP biosynthesis. TOC Graphic O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=104 SRC="FIGDIR/small/739211v1_ufig1.gif" ALT="Figure 1"> View larger version (25K): org.highwire.dtl.DTLVardef@8cabdorg.highwire.dtl.DTLVardef@1364ba5org.highwire.dtl.DTLVardef@1a059f4org.highwire.dtl.DTLVardef@71c9fc_HPS_FORMAT_FIGEXP M_FIG C_FIG

biochemistry↗

bioRxiv2026

Concurrent Stereotactic Body Radiation Therapy and KRAS Inhibition Synergistically Improve Pre-clinical Pancreatic Cancer Treatment

Pancreatic ductal adenocarcinoma (PDAC) is one of the most challenging cancers to treat due to the dismal survival rate, poor post-treatment outcome and profound resistance to a wide range of therapies. With mutant KRAS being a key driver, small molecule inhibitors targeting KRAS or pan-RAS (KRASi) have demonstrated exciting preclinical and early clinical anti-tumor efficacy, and the pan-RAS(ON) inhibitor daraxonrasib (RMC-6236) recently achieved Phase 3 clinically meaningful improvements in patient survival compared to chemotherapy. But resistance to RAS/KRAS inhibitor inevitably develops, which limits and compromises the treatment outcome. In this study, we investigated the combination of stereotactic body radiation therapy (SBRT) and KRAS inhibition (MRTX1133 and daraxonrasib) in the treatment of preclinical PDAC models. We found that this combination strategy synergistically suppresses PDAC cell growth in vitro and enhances tumor control while minimizing local recurrence in orthotopically implanted KPC (LSL-KrasG12D/+;Trp53R172H/+;Pdx1-Cre) murine PDAC tumors in vivo. As radiation therapy (RT) induces ferroptosis in multiple cancer types and mutant KRAS promotes various anti-ferroptotic mechanisms, we tested the role of ferroptosis in promoting tumor-control efficacy. Intriguingly, the addition of a ferroptosis inhibitor, liproxstatin-1, to the combination therapy significantly abrogated the in vivo synergism between SBRT and KRAS inhibition, suggesting that treatment-induced ferroptosis at least partially drives the synergistic efficacy of this combination strategy. Our study indicates that this SBRT-KRASi combination has the potential to overcome treatment resistance and improve outcomes in PDAC patients. These data directly support the design of a planned multi-center Phase 2 clinical trial with this combination strategy in locally advanced PDAC.

cancer biology↗

bioRxiv2026

A pesticide hijacks a neurohormonal circuit to block ovulation in insects

The global decline of insect populations threatens ecosystem stability, yet the hidden physiological impacts of sublethal pesticide exposure remain largely uncharted. Here we show that emamectin benzoate (EB), a widely used insecticide, suppresses female fertility not by disrupting egg development, but by specifically blocking ovulation. Through a systematic genetic screen in Drosophila, we identify the glutamate-gated chloride channel GluCl in a discrete set of octopaminergic neurons as the primary neural target. EB subverts the activity of this circuit, thereby impairing octopamine-dependent follicle rupture, suppressing ovarian ecdysteroid signaling, and altering oviduct muscle dynamics--three coordinated processes essential for egg release. Strikingly, this ovulatory blockade is recapitulated in diverse dipteran pests and disease-vector mosquitoes, revealing a conserved vulnerability. Our findings establish a neurohormonal mechanism by which a common agrochemical hijacks reproductive control, and they expose ovulation as a critical, hitherto unrecognized nexus between environmental chemicals and insect population dynamics. SignificanceSublethal pesticide exposure can profoundly alter insect physiology beyond acute toxicity, yet the underlying mechanisms remain unclear. Here, we show that emamectin benzoate (EB), a widely used avermectin insecticide, suppresses ovulation and female fecundity in Drosophila by disrupting octopaminergic signaling, follicle rupture, and oviduct muscle contraction. Mechanistically, EB acts through GluCl-dependent neural pathways to impair ovulation-related processes and reproductive muscle dynamics. Notably, these inhibitory effects are conserved across multiple dipteran species, suggesting that reproductive suppression represents a broader physiological consequence of EB exposure. Our findings reveal a previously unrecognized neuro-reproductive mechanism of pesticide action and provide new insight into how sublethal insecticide exposure may influence insect population dynamics.

neuroscience↗

bioRxiv2026

A CSF Proteomic Clock Reveals Opposing Brain-Aging Programs and Predicts Neurological Disease Progression

Aging is the risk factor for neurological disease, yet the molecular architecture of human brain aging remains poorly defined. Here, we analyzed more than 10,000 cerebrospinal fluid (CSF) proteomes across multiple cohorts and proteomic platforms to developed a 249-protein CSF aging clock that accurately predicts age and generalizes across independent datasets. CSF brain-age acceleration was increased across diverse neurological diseases, associated with blood-brain barrier (BBB) dysfunction, and predictive of longitudinal cognitive decline, neuroimaging progression, and dementia conversion. A simplified 30-protein panel retained similar prognostic performance. Biologically, clock proteins resolved two opposing programs: pro-aging activation of immune, vascular/BBB, ECM and coagulation pathways marked by CHI3L1, CD14, VWF, LRG1 and LTBP2, and anti-aging collapse of neuronal, synaptic, axonal and structural-maintenance programs marked by NPTX2, COL1A2, NID1, CDH8 and PENK. Brain-wide single-cell and regional mapping linked these programs to disease-vulnerable compartments. These findings establish a molecular framework for biological brain aging and neurological disease vulnerability.

neuroscience↗

bioRxiv2026

Ginsenoside Ro ameliorates diabetic cardiomyopathy by maintaining ciliary homeostasis and enhancing antioxidation

ObjectiveTo investigate and clarify the role of Ginsenoside Ro (GRo) in diabetic cardiomyopathy (DiaCM) and to elucidate the molecular mechanism by which GRo ameliorates DiaCM. Methods[circled1] The construct of type 2 diabetic mouse model. The bought C57BL/6 male mice were housed in a specific pathogen-free (SPF) animal facility and randomly divided into control, STZ (model), STZ + GRo, and control+GRo groups. The STZ (model) and STZ + GRo groups were fed a high-fat and high-glucose diet combined with intraperitoneal injection of streptozotocin (STZ). The control and control + GRo groups were fed a normal diet, while the control + GRo and STZ + GRo groups were treated with GRo via oral gavage. Then, all groups were evaluated for cardiac function and structure by small animal echocardiography and histological staining including hematoxylin and eosin (HE) and Massons trichrome staining to screen and confirm diabetic cardiomyopathy in mice. Finally, immunofluorescence staining of cilia in mouse heart tissue was performed to determine whether GRo inhibits abnormal ciliary growth. [circled2] The construct of cell models. First, the CCK-8 (Cell Counting Kit-8) assay was used to separately evaluate the cytotoxicity of GRo and the combination of TGF-{beta}1 and PA in myocardial fibroblasts and cardiomyocytes. Subsequently, mouse myocardial fibroblast lines (MCFs) were treated with transforming growth factor-beta 1 (TGF-{beta}1), and H9c2 cardiomyocytes were treated with palmitic acid (PA). Both cell types then received the GRo treatment. [circled3] Molecular and cellular testing. Firstly, we measured serum levels of cardiac injury markers (CK-MB, MYO, and TNNI3), glutathione (GSH), and malondialdehyde (MDA). Secondly, we examined the expression of myocardial fibrosis-related genes (Col1a1, etc.), myocardial hypertrophy markers (Nppa, etc.), cilia-specific genes (Pkd1, etc.), and oxidative stress-related genes (Nrf2, etc.) in both animal and cell samples by Western blotting and RT-qPCR. Finally, we used immunofluorescence staining of myocardial fibroblasts to detect cilia length and phalloidin staining of cardiomyocytes to measure their cross-sectional area. [circled4] The correlation mechanism. Firstly, the cilia-specific inhibitory drug HIP-4 was used to disrupt cilia homeostasis by inhibiting cilia growth. Secondly, small activating RNA (saRNA) was used to upregulate the Pkd1 gene to verify whether GRo exerts its anti-fibrotic effects through the inhibition of PC1. Results[circled1] Animal level. A diabetic cardiomyopathy mouse model was successfully established by combining STZ injection with a high-fat and high-glucose diet, and treatment with GRo significantly ameliorated the associated symptoms. [circled2] Cellular level. We successfully established a myocardial fibrosis model by treating myocardial fibroblasts with TGF-{beta}1, and a myocardial hypertrophy model by treating cardiomyocytes with PA. Immunofluorescence staining demonstrated that GRo significantly decreased cilia length in the fibrosis model, while phalloidin staining showed that GRo significantly attenuated the increase in cardiomyocyte cross-sectional area. [circled3] Molecular level. Compared with the model group, GRo treatment significantly reduced serum levels of cardiac injury markers (CK-MB, MYO and TNNI3), glutathione (GSH) and malondialdehyde (MDA). Western blotting and RT-qPCR analyses of both animal and cell samples revealed that GRo markedly alleviated indicators of myocardial fibrosis and hypertrophy, while also suppressing cilia-specific genes and oxidative stress-related genes. Overall, GRo significantly ameliorated the markers associated with myocardial fibrosis and hypertrophy, and inhibited cilia-specific protein expression as well as oxidative stress parameters. [circled4] The correlation mechanism. The cilia-specific drug hedgehog pathway inhibitor 4 (HPI-4) was used to revealed that cilia homeostasis is closely linked to myocardial fibrosis and shortened cilia inhibit the fibrosis progression. Furthermore, upregulation of the Pkd1 gene by small activating RNA demonstrated that PC1 overexpression abrogates the therapeutic effect of GRo. Finally, GRo can alleviate DiaCM.

Molecular Biology↗