The global outbreak of hand, foot, and mouth disease (HFMD), mainly caused by enterovirus A71 (EV71) and coxsackievirus A16 (CA16) of the genus enterovirus, poses a serious threat to the health of young children, and severe neurological manifestations and associated symptoms are primarily attributed to EV71 infection. There is currently no specific antiviral drug available for HFMD. In this study, we identified Moracin M (MM), a flavonoid compound derived from Smilax china, as a potent inhibitor of EV71 and CA16 infection. Mechanistic investigation revealed that MM exerts its antiviral effects primarily by inhibiting viral attachment to host cells by targeting the virus. Furthermore, MM showed excellent safety and significant anti-EV71 activity in neonatal mice, and it also effectively suppressed the inflammatory response and alleviated muscle, lung, and brain tissue damage induced by EV71 infection. In conclusion, our study identified and demonstrated MM as a promising inhibitor of EV71 and CA16, providing important insights for the further development of MM as a potential therapeutic agent for HFMD caused by these viruses.

HIV-1 infection activates microglia and triggers neuroinflammation, which is the primary cause of HIV-associated neurological complications. Our previous study demonstrated that HIV-1 infection upregulates m6A modification in microglia, and in this study we investigated the underlying regulatory mechanisms. Through viral protein screening, we identified Vpr as the protein responsible for increasing m6A modification. Further analysis revealed that HIV-1 infection reduces the level of the m6A demethylase ALKBH5. Vpr deleted HIV-1 infection and Q65R mutant Vpr expression experiments demonstrated that Vpr is capable of degrading ALKBH5 protein via the ubiquitin-proteasome pathway by interacting with ALKBH5. Addition of m6A inhibitors or overexpression of ALKBH5 inhibited Vpr-induced microglial activation and the production of inflammatory cytokines, suggesting that the upregulation of m6A modification might play a crucial role in microglial activation induced by Vpr. As microglial activation is a major cause of neuroinflammation leading to neuronal damage, this study provides new insights for understanding the interactions between HIV-1 and microglia, and might provide new ideas for the prevention strategies study on the neuroinflammation caused by HIV-1 infection.

Cognitive resilience in Alzheimer's disease (AD) requires the maintenance of synaptic integrity despite progressive pathological insults. Reactive astrocytes can switch between neuroprotective and neurotoxic states, and their maladaptive transition significantly accelerates neurodegeneration, yet the molecular drivers of this shift remain elusive. Here, using published single-nucleus transcriptomic data, we identified the sialyltransferase St6galnac5 as a candidate regulator associated with reactive, pro-inflammatory astrocyte states. We further show that astrocyte-specific, AAV-mediated knockdown of St6galnac5 in female 3xTg-AD mice improves spatial learning, memory and anxiety-like behaviors. Neuropathological assessment revealed that this functional recovery was underpinned by a marked reduction in amyloid-β and tau pathologies, alongside the preservation of synaptic integrity. Consistent with a shift toward a less inflammatory astrocyte state, St6galnac5 knockdown decreased A1-associated markers and increased A2-associated markers in vitro and alleviated neurite outgrowth deficits in neuron-astrocyte co-culture. Together, our findings identify St6galnac5 as a critical molecular switch driving astrocytic dysfunction in AD, and further propose that targeted inhibition of this sialylation pathway represents a viable strategy to bolster astrocytic resilience and slow disease progression.

Two new coumarins (1 and 2) and two new chromones (3a and 3b) were isolated from Gerberapiloselloides, and their structures were elucidated by 1D and 2D NMR, HRESIMS, X-ray diffraction, experimental and calculated ECD. All of the isolates were evaluated for the anti-neuroinflammatory effect on LPS induced BV2 cells. As a result, compounds 1 and 2 down-regulated the mRNA level of TNF-alpha and IL-1 beta detected by qPCR. Taken together, 1 and 2 might possess the potential to the development of lead compound for the neuroinflammation treatment.

Beta-hemoglobin (HBB) is expressed in neurons, though its function remains incompletely understood. Prenatal stress (PS) is a well-established risk factor for neuropsychiatric disorders; however, the molecular mechanisms underlying its long-term effects are still unclear. In this study, we demonstrated that PS induces persistent anxiety- and depression-like behaviors in adult rat offspring, along with a significant downregulation of HBB in prefrontal cortical neurons. Deficiency of HBB resulted in neuronal hypoxia, impairing dendritic development, synaptic function, and neuronal connectivity; whereas overexpression of HBB rescued these behavioral and neuronal deficits. In healthy control rat offspring, knockdown of HBB in prefrontal cortical neurons recapitulated the phenotypes induced by PS. To intervene in these pathogenic processes, we found that early-life hyperbaric oxygen therapy (HBOT) restored HBB expression, alleviated neuronal hypoxia, and prevented the development of affective disorders in adulthood. Our findings identify cortical HBB as a key mediator linking PS to neurodevelopmental impairments and suggest hyperbaric oxygen therapy as a potential therapeutic strategy for stress-related psychiatric disorders.

Alzheimer's disease (AD) research has primarily focused on amyloid beta (Aβ) and tau protein; however, drug development targeting these two proteins has been disappointing. Therefore, there is an urgent need to explore the novel pathogenic mechanisms underlying AD. Recently, we found that expression of the K670N/M671L-mutated amyloid precursor protein (APP) in 293T cells significantly reduced membrane ferroportin (FPN) levels. Furthermore, 2-month-old APP/PS1 mice exhibited a marked decrease in membrane FPN levels, while total FPN expression and Aβ levels remained unchanged. Further studies revealed that features of ferroptosis were present in the brains of 2-month-old APP/PS1 mice, and that treatment with ferroptosis inhibitors or iron chelation significantly alleviated early pathological changes and cognitive impairment in these animals. In addition, supplementation with an APP-FPN binding peptide during the early phase ameliorated AD-related pathologies, including Aβ deposition, neuroinflammation, oxidative stress, and synapse-associated protein deficits, in APP/PS1 mice. Collectively, our findings suggest that APP mutations may contribute to early brain pathological changes and subsequent memory impairment in AD by downregulating membrane trafficking of FPN and inducing ferroptosis, thereby providing new molecular targets for drug development.

High mobility group box 1 (HMGB1), when released extracellularly, plays a pivotal role in the development of spinal cord synapses and exacerbates autoimmune diseases within the central nervous system. In experimental autoimmune encephalomyelitis (EAE), a condition that models multiple sclerosis, the levels of extracellular HMGB1 and interleukin-33 (IL-33) have been found to be inversely correlated. However, the mechanism by which IL-33 deficiency enhances HMGB1 release during EAE remains elusive. Our study elucidates a potential signaling pathway whereby the absence of IL-33 leads to increased binding of P300/CBP-associated factor with HMGB1 in the nuclei of astrocytes, upregulating HMGB1 acetylation and promoting its release from astrocyte nuclei in the spinal cord of EAE mice. Conversely, the addition of IL-33 counteracts the TNF-α-induced increase in HMGB1 and acetylated HMGB1 levels in primary astrocytes. These findings underscore the potential of IL-33-associated signaling pathways as a therapeutic target for EAE treatment.

Brain damage induced by ischemia promotes the development of cognitive dysfunction, thus increasing the risk of dementia such as Alzheimer's disease (AD). Studies indicate that cellular acidification-triggered activation of asparagine endopeptidase (AEP) plays a key role in ischemic brain injury, through multiple molecular pathways, including cleavage of its substrates such as SET (inhibitor 2 of PP2A, I2 PP2A) and Tau. However, whether direct targeting AEP can effectively prevent post-stroke cognitive impairment (PSCI) remains unanswered. Here, we explored the therapeutic effect and underlying mechanism of the AEP inhibitor AENK on cognitive impairment of the rats with middle cerebral artery occlusion (MCAO) and on neuronal damage in cultured primary neurons exposed to oxygen and glucose deprivation (OGD). We found that the administration of AENK significantly reduces activated AEP levels in ischemic rat brains, attenuates cognitive deficits, and rescues synaptic dysfunction. For the mechanism, with AEP inhibition, cleavage of SET, inhibition of protein phosphatase 2A (PP2A), and Tau hyperphosphorylation resulted from PP2A inhibition, were all completely or partially reversed. In primary neurons, AENK effectively prevents AEP activation, SET cleavage and cytoplasmic retention, tau hyperphosphorylation and synaptic damage induced by OGD. We conclude that AENK ameliorates cognitive impairment and prevents tau hyperphosphorylation, through inhibiting AEP-mediated cleavage of SET in ischemic brain injury, and direct inhibition of AEP might be a potential therapeutic strategy for preventing synaptic damage and cognitive impairment after stroke.

Aberrant phosphorylation of the Tau protein represents a critical event in the pathogenesis of Alzheimer's disease (AD); however, therapeutic interventions specifically targeting this modification remain limited. Therefore, a thorough understanding of the molecular mechanisms underlying Tau hyperphosphorylation is essential for the development of effective preventive and therapeutic strategies against AD. The RNA-binding protein MUSASHI1 (MSI1) is recognized for its significant role in neurodevelopment, and previous studies have reported its dysregulated overexpression in the brains of AD patients. In the current investigation, we demonstrate that MSI1 expression progressively increases in parallel with the advancement of Tau pathology in P301S transgenic mouse models. Furthermore, our findings suggest that MSI1 activates the p38 mitogen-activated protein kinase (MAPK) signaling pathway, thereby promoting Tau phosphorylation. Additionally, we have identified two microtubule-associated proteins as novel potential interaction partners of MSI1 within neuronal cells. Collectively, these results reveal a previously uncharacterized mechanism that may contribute to aberrant Tau phosphorylation in AD, offering new directions for future research in this field.

High-salt (HS) diet is an established risk factor for cognitive impairment, but the underlying mechanisms remain unclear. This study reveals that HS diet reduces SHANK1, a key postsynaptic scaffolding protein, via downregulation of the PKA/CREB pathway, leading to synaptic dysfunction and cognitive deficits in rats. RNA sequencing of HS-fed rat hippocampi showed downregulation of cAMP signaling and SHANK1 expression. Pharmacological inhibition of PKA/CREB reduced SHANK1 levels and impaired dendritic structure and synaptic function, while PKA activation restored CREB activity and SHANK1 expression, reversing HS-induced deficits. Notably, CREB activation is essential for SHANK1 regulation, as a CREB mutant (S133A) blocked the effects of PKA activation, and a constitutively active CREB (S133D) prevented SHANK1 downregulation. These findings highlight the PKA/CREB/SHANK1 pathway as a potential therapeutic target for HS-induced cognitive dysfunction.

BackgroundThe association between serum iron concentration and cognitive ageing remains inconsistent, particularly in rural Chinese populations where both iron deficiency and excess may coexist.MethodsIn this cross-sectional analysis of 737 community-dwelling adults aged ≥60 years from rural Taiyuan, China, cognitive impairment was assessed with the Chinese-language Mini-Mental State Examination (MMSE) using education-specific cut-offs. Serum iron was quantified by inductively coupled plasma-mass spectrometry and categorized into tertiles (low, medium, high). Multivariable logistic regression models adjusted for sociodemographic, lifestyle and clinical covariates examined independent and joint associations; restricted cubic splines (RCS) evaluated dose–response patterns.ResultsAfter comprehensive adjustment, serum iron demonstrated a non-linear relationship with cognitive impairment risk. Relative to the lowest tertile, medium (OR = 0.61, 95% CI: 0.39–0.95) and high (OR = 0.59, 95% CI: 0.37–0.93) serum iron concentrations were associated with reduced odds of cognitive impairment. Spline analysis suggested a U-shaped relationship: risk decreased with rising serum iron up to a nadir and then trended upward; however, the departure from linearity was not statistically significant (P-nonlinear = 0.194).ConclusionBoth low and high serum iron levels are linked to poorer cognitive performance in this rural cohort, supporting the need to maintain iron within an optimal range for healthy cognitive ageing.

Alzheimer’s disease (AD) is a complex neurodegenerative disorder with multifaceted pathogenesis, which has been extensively investigated, yet effective treatments remain lacking. Splicing factor proline and glutamine rich (SFPQ) is known to play a crucial role in neurodegenerative diseases, including antioxidant-related functions and regulating gene expression within brain neurons. However, the specific role of SFPQ in AD pathology is not well understood. In this study, an AD mouse model was established through lateral ventricular injection of amyloid-beta 1–42 (A β 1–42 ). Subsequently, adeno-associated virus was administered to overexpress SFPQ in the hippocampus of AD mice. The results demonstrate that SFPQ overexpression improves recognition and memory in AD mice, while reducing AD-related marker proteins such as amyloid precursor protein (APP) and Tau. Additionally, synaptic and memory-associated proteins, as well as antioxidant proteins like glutathione S-transferase (GST) and heme oxygenase-1 (HO-1), were upregulated. The ratio of antiapoptotic protein Bcl-2 to proapoptotic protein Bax also increased. Furthermore, phosphorylated phosphoinositide 3-kinase (p-PI3K)/PI3K and phosphorylated protein kinase B (p-AKT)/AKT ratios were elevated, indicating activation of the PI3K/AKT signaling pathway. These findings suggest that SFPQ may serve as a promising molecular target for the prevention and treatment of AD.

Background Post‐stroke depression (PSD) is a prevalent neuropsychiatric complication of stroke. However, the mechanisms underlying PSD are still unclear. Here, we aimed to investigate the role of HCN1 (hyperpolarization‐activated cyclic nucleotide‐gated cation channel 1) in the pathogenesis of PSD and its underlying mechanisms. Methods The PSD mice model was established by middle cerebral artery occlusion in vivo. Four weeks after middle cerebral artery occlusion, anxiety‐ and depression‐like behaviors of mice were evaluated by various behavioral tests. HCN channels were downregulated by pharmacological inhibitor or neuron‐specific adeno‐associated virus. The oxygen–glucose deprivation/reoxygenation model in SY5Y cells was used to study the pathogenesis of PSD in vitro. Results Mice exhibited anxiety‐ and depression‐like behavior 4 weeks after middle cerebral artery occlusion, along with a significant increase in HCN1 protein expression in the ischemic hippocampus. Furthermore, the Ih current on neurons in the hippocampus was notably enhanced, whereas neuronal excitability was decreased in PSD mice. Treatment with HCN channel selective inhibitor ZD7288 protected SY5Y cells against oxygen–glucose deprivation/reoxygenation injury by suppressing K+ efflux. Additionally, we observed a significant increase in protein expressions of NLRP3 (nucleotide‐binding domain‐like receptor protein 3) inflammasome pathway‐related molecules in the ischemic hippocampus of PSD mice. Knockdown of HCN1 channels via virus injection into the hippocampus resulted in decreased protein expressions of NLRP3 inflammasome‐related molecules and improvement in anxiety‐ and depression‐like behaviors in PSD mice. Conclusions Downregulation of HCN1 channels has a beneficial effect on PSD by suppressing the NLRP3 inflammasome pathway, thus offering promise as a strategy for preventing and treating PSD.

One previously undescribed 2-arylbenzofuran derivative (1), three stilbenes (2-4), three acetophenone glycosides (5-7), three phenylpropanoids (8-10), and eight flavonoids (11-18) were isolated from the rhizome of Smilax china L. in this study. NMR spectra elucidated their structures and were compared with previously reported data. Compounds 1, 5-7, 10, and 18 were identified for the first time in Smilax china L. The cytotoxicity of the isolated compounds was detected on HeLa cells by the CCK-8 method, and compound 4 was found to possess the highest cytotoxic activity with an IC50 value of 34.81 +/- 1.50 mu g/mL. Further studies revealed that the cytotoxic effect of compound 4 on HeLa cells was related to apoptosis and the cell cycle. In addition, the chemotaxonomic significance of the isolated compounds was discussed.

Adult neurogenesis plays a crucial role in maintaining brain homeostasis and can respond to neurogenic injuries. However, the adult mammalian spinal cord has extremely limited intrinsic neurogenic ability. Here, we show that in vivo astrocyte-to-neuron conversion can regenerate functional neurons after spinal cord injury (SCI) through CRISPRa-mediated activation of endogenous transcription factors Ngn2 and Isl1. Lineage tracing confirms that the origin of these induced neurons is reactive astrocytes, rather than endogenous neurons. Furthermore, these induced neurons express specific markers of motor neurons and glutamatergic neurons and form synaptic connections with ascending and descending spinal pathways. Importantly, astrocyte-to-neuron conversion promotes propriospinal axon regeneration, improves the neuromuscular junction (NMJ) morphology and function of muscle, and finally promotes motor functional recovery after SCI. In summary, our results would contribute to resolving the controversy surrounding lineage reprogramming and demonstrate that in vivo cell conversion may be a potential therapeutic strategy for treating SCI.

Two novel phenylpropanoids (compounds 1 and 2) and 11 known compounds were isolated from Smilax china L. Their structures were determined by NMR (1D and 2D) and high-resolution electrospray ionization mass spectrometry. Further, the cytotoxic activity of all the isolated compounds against HeLa, 4T1, and U251 tumor cells was evaluated using the cell counting kit-8 assay, revealing that compound 13 showed significant cytotoxicity toward HeLa cells. Further investigations explored the impact of compound 13 on the mitochondrial membrane potential, concentration of reactive oxygen species, wound-healing distance, and cell cycle of HeLa cells. Notably, compound 13 significantly decreased mitochondrial membrane potential, suppressed cell migration, and increased intracellular reactive oxygen species levels in HeLa cells. Furthermore, compound 13 inhibited HeLa cell-cycle progression in the S phase. These findings indicate that compound 13 is a potential drug lead for the treatment of cervical cancer.

Ambient fine particulate matter (PM2.5) is a threat to public health. The P2 X 7purinergic receptor (P2X7R) is a modulator that responds to inflammation. Yet the role of P2X7R in the mediation of PM2.5-induced pulmonary cytotoxicity is rarely investigated. In this study, the expression of P2X7R and its effect on cell viability, oxidative damage, apoptosis, mitochondrial dysfunction and underlying mechanism following PM2.5 treatment in rat alveolar macrophages (NR8383) were analyzed. The outcome indicated that PM2.5 exposure significantly increased the expression of P2X7R, while P2X7R antagonist oATP markedly alleviate the production of reactive oxygen species (ROS), Nitrite Oxidation (NO), mitochondrial membrane potential, apoptosis rate, and release of inflammatory cytokines. In contrast, P2X7 agonist BzATP showed opposite effect in PM2.5-treated NR8383 cells. Therefore, these results demonstrated that P2X7R participated in PM2.5-induced pulmonary toxicity, while the blockade of P2X7R is a promising therapeutic approach of treating PM2.5-induced lung diseases.

AimsThis study was designed to investigate the role of growth arrest and DNA damage-inducible β (GADD45B) in modulating fear memory acquisition and elucidate its underlying mechanisms.Main methodsAdeno-associated virus (AAV) that knockdown or overexpression GADD45B were injected into ventral hippocampal CA1 (vCA1) by stereotactic, and verified by fluorescence and Western blot. The contextual fear conditioning paradigm was employed to examine the involvement of GADD45B in modulating aversive memory acquisition. The Y-maze and novel location recognition (NLR) tests were used to examine non-aversive cognition. The synaptic plasticity and electrophysiological properties of neurons were measured by slice patch clamp.Key findingsKnockdown of GADD45B in the vCA1 significantly enhanced fear memory acquisition, accompanied by an upregulation of long-term potentiation (LTP) expression and intrinsic excitability of vCA1 pyramidal neurons (PNs). Conversely, overexpression of GADD45B produced the opposite effects. Notably, silencing the activity of vCA1 neurons abolished the impact of GADD45B knockdown on fear memory development. Moreover, mice with vCA1 GADD45B overexpression exhibited impaired spatial cognition, whereas mice with GADD45B knockdown did not display such impairment.SignificanceThese results provided compelling evidence for the crucial involvement of GADD45B in the formation of aversive memory and spatial cognition.

In the process of searching for anti-breast cancer agents, five sesquiterpene lactones (1-5), including two previously undescribed ones, yjaponica B-C (1-2), were isolated from the herb of Youngia japonica. Their structures were elucidated by spectroscopic data analyses and Marfey's method. Cytotoxic activities of all compounds against A549, U87, and 4T1 cell lines were tested using the CCK8 assay. The result showed that compound 3 possessed the highest cytotoxic activity against 4T1 cells with an IC50 value of 10.60 μM. Furthermore, compound 3 distinctly induced apoptosis, inhibited immigration, and blocked the cell cycle of 4T1 cells. In addition, compound 3 induced the production of reactive oxygen species. Further anticancer mechanism studies showed that compound 3 significantly upregulated expression of the cleaved caspase 3 and PARP, whereas it downregulated the expression of Bcl-2, cyclin D1, cyclin A2, CDK4, and CDK2. Taken together, our results demonstrate that compound 3 has a high potential of being used as a leading compound for the discovery of new anti-breast cancer agent.