Green environments have long been recognized to produce calming, anxiolytic effects, although the underlying neural mechanisms remain poorly understood. Here, we find that green light treatment can relieve anxiety-like behaviors by activating a neural pathway from the medial part of the secondary visual cortex (V2M) to the prelimbic cortex (PrL) in mice subjected to acute stress. Viral tracing, fiber photometry, and microendoscopic calcium imaging together show that green light activates glutamatergic inputs from V2M (V2MGlu) to inhibitory neurons expressing corticotropin-releasing factor in the PrL (PrLCRF), which in turn suppress local glutamatergic neurons (V2MGlu→PrLCRF→Glu). Chemogenetic activation of this pathway mimics the anxiolytic effects of green light, whereas artificial inhibition of this pathway or pharmacological blockade of CRF receptor 1 (CRFR1) in the PrL abolishes green-light-induced anxiolysis in acute stress model mice. This study thus indicates a CRF-dependent neural mechanism underlying the anxiolytic effects of green light under stress conditions.

Pharmacological interventions can reduce immediate headache, but long-term effectiveness and suitability for chronic migraine remain limited. Although aromatherapy with lavender essential oil (LEO) reportedly alleviates headaches, the neural mechanisms remain uncertain. Here, we found that LEO inhalation exerts antinociceptive effects by activating orexinergic neurons in the lateral hypothalamus (LHOX) of mice with nitroglycerin-induced chronic migraine. Viral tracing, in vivo fiber photometry, and behavior assays revealed that LEO activates glutamatergic inputs from the anterior olfactory nucleus (AONGlu) to LHOX neurons, which subsequently inhibit the lateral parabrachial nucleus expressing calcitonin gene-related protein (LPBNCGRP) by activating local GABAergic neurons in LPBN (LPBNGABA) through orexin receptor type 2 (OX2R) signaling, ultimately alleviating chronic migraine-like behaviors. Chemogenetic activation of the AONGlu→LHOX→LPBNGABA→CGRP circuit mimics LEO-induced antinociception, while its inhibition or pharmacological OX2R blockade abolishes this effect in migraine mouse models. This study thus uncovers an orexinergic mechanism underlying the antinociceptive effects of LEO.

Despite its long-recognized effects in relieving pain, the neural substrates of auricular stimulation remain elusive. Here, we show that trans-auricular vagus nerve stimulation (taVNS), i.e., electrical stimulation of the auricular concha, effectively induces analgesia in a mouse model of neuropathic pain. Viral tracing, microendoscopic calcium imaging, and multi-electrode recordings reveal that auricular vagal signals travel to the jugular-nodose ganglia (JNG), which in turn connect to pro-opiomelanocortinergic neurons in nucleus tractus solitarius (NTS), subsequently activating glutamatergic neurons in ventrolateral periaqueductal gray (vlPAG). Optogenetic stimulation of central vagus terminals, JNG-derived auricular peripheral fibers, or vlPAG-projecting NTS neurons mimics taVNS-induced analgesia, whereas chemogenetic silencing of central vagus terminals or NTS neurons abolishes this effect. This study identifies an auricle-to-brain circuit underlying taVNS-driven analgesia in mice, with potential for facilitating taVNS optimization for pain management and other neurological conditions.

Background/Objectives: Colorectal cancer (CRC) is a major cause of cancer- related deaths worldwide and poses a significant global health challenge. Metabolic reprogramming, including transketolase (TKT) upregulation in the pentose phosphate pathway, is a hallmark of CRC. Drugs targeting TKT have shown promise in cancer treatments; however, their mechanisms are poorly understood. Methods: In this study, by integrating bioinformatics analysis of 494 cases from the TCGA database, we identified that transketolase (TKT) may be a potential key target for colorectal cancer. Furthermore, we systematically screened the Natural Product Library for HTS (L6000) TargetMol using molecular docking technology. Results: Anamorelin was identified, which exhibited optimal spatial matching with the ligand in the target protein crystal structure. As a selective growth hormone secretagogue receptor (GHS-R) agonist, anamorelin has been clinically demonstrated to significantly improve appetite and increase lean body mass in patients with CRC by activating the GHS-R signaling pathway and is currently used for treating cancer cachexia in patients with advanced CRC. However, existing research has primarily focused on its metabolic benefits, and its direct effects on tumor cell proliferation, migration, and related molecular mechanisms remain unclear. This study elucidated the regulatory relationship between anamorelin and TKT in CRC, thereby addressing a critical knowledge gap in this field (Fig. 1). Discussion: These findings reveal a previously unrecognized antitumor mechanism of anamorelin through TKT inhibition. These findings provide preliminary evidence of a TKT-associated mechanism of anamorelin in CRC, offering dual benefits in cachexia management and tumor suppression, and warrant further validation in advanced preclinical models. Further preclinical and clinical validation is warranted to translate these insights into practice. Conclusion: By integrating bioinformatics analysis with in vitro cellular experiments, we systematically validated the molecular mechanism whereby anamorelin inhibited tumor cell proliferation and migration via the targeted suppression of TKT. These findings provide a theoretical foundation for the development of novel metabolism-based therapeutic strategies for CRC, ultimately contributing to improved clinical outcomes.

Chronic stress can induce pain and drive development of aversion, although the specific neural circuits that contribute to encoding such behavioral responses remain unclear. Here, we find that noradrenergic neurons in locus coeruleus (NELC) regulate the formation of allodynia and aversion under chronic restraint stress in mice, driven by enhanced activity of NELC-projecting GABAergic neurons in the central amygdala (CeA). Selectively inhibiting this GABAergic CeA→LC pathway significantly alleviates pain sensitization and aversion behaviors, but without impacting chronic stress-associated depression-like behaviors. Additionally, these NELC neurons receiving inputs from CeA monosynaptically activate glutamatergic neurons in the lateral periaqueductal gray (GluLPAG) to mediate pain sensitization and aversion. This study thus defines a specific CeA→LC→LPAG pathway through which chronic stress leads to pain and aversion, without affecting depression-like behaviors, expanding our fundamental understanding of the circuit basis for stress-related pain.

The central nervous system regulates glandular secretions by modulating visceral organ functions. However, the anatomical and functional connections between the brain and digestive enzyme-producing pancreatic acinar cells remain poorly defined. Using the hypertriglyceridemia-associated acute pancreatitis (HTGP) mouse model, we aimed to describe a functional transneuronal circuit connecting the hypothalamus to pancreatic acinar cells in mice. This circuit originates from a subpopulation of oxytocin neurons in the paraventricular hypothalamic nucleus (PVNOXT), and reaches the exocrine pancreas via acetylcholinergic neurons in the dorsal motor nucleus of the vagus (DMVACh) to innervate acinar cells. Silencing of PVNOXT neurons suppresses digestive enzyme secretion and inhibits the inflammatory response in HTGP. Conversely, stimulation of these neurons induces inflammation by dysregulating secretory pathways in acinar cells. Single-cell RNA sequencing revealed that WD repeat and FYVE domain-containing 1 (Wdfy1), a gene expressed in the OXT neuron subpopulation, plays a critical role in the acinar cell-mediated inflammatory response, and its function is essential for the PVNOXT-DMVACh axis.

Although pain sensitization is among the most common conditions affecting the elderly, its underlying neural mechanisms remain unclear. Here, we found that mechanical hypersensitivity arises from enhanced glutamatergic excitability in primary somatosensory cortex due to bradykinin receptor B2 (BDKRB2) upregulation by dystrophic microglia during aging. Specifically, in vivo fiber photometry in aged mice revealed hyperactivity of glutamatergic neurons in primary somatosensory cortex of hindlimb (S1HLGlu), while chemogenetic inhibition of these neurons reverses pain hypersensitivity in aged mice. BDKRB2 expression on S1HLGlu neurons is significantly increased in aged mice, whereas its conditional knockdown restores pain sensitization, and its overexpression leads to nociceptive hypersensitivity in young mice. Moreover, chemogenetic or pharmacological inhibition of dystrophic microglia in aged mice reduces BDKRB2 levels, alleviating pain hypersensitivity. The present study thus demonstrates that microglia-mediated S1HLGlu hyperactivity drives development of aging-related pain via BDKRB2 signaling, suggesting several potentially effective therapeutic targets for treating geriatric pain.

The neural mechanisms underlying sex-specific pain, in which males and females exhibit distinct responses to pain, remain poorly understood. Here we show that in a mouse model of male-specific pain hypersensitivity response to pain conditioning environments (contextual pain hypersensitivity model), elevated free-testosterone leads to hyperactivity of glutamatergic neurons in the medial preoptic area (GlumPOA) through activation of androgen receptor signaling, which in turn induces contextual pain hypersensitivity in male mice. Although not observed in naïve female mice, this pain phenotype could be induced in females via chronic administration of testosterone propionate. In addition, GlumPOA neurons send excitatory inputs to GABAergic neurons in the ventrolateral periaqueductal gray (GABAvlPAG) that are required for contextual pain hypersensitivity. Our study thus demonstrates that testosterone/androgen receptor signaling enhances GlumPOA → GABAvlPAG pathway activity, which drives a male-specific contextual pain hypersensitivity, providing insight into the basis of sexually dimorphic pain response.

Pain in one part of the body profoundly diminishes the sensation of pain in other parts of the body in humans. Here, we found that pain-related behaviors in hindpaw are inhibited by noxious stimuli from diverse body regions in mice. Using activity-dependent cell labeling in male FosTRAP2 mice, we captured a neuronal ensemble in the layers 2-4 of secondary somatosensory cortex (S2) that was activated during pain at diverse body regions induced analgesia. Single-cell projection analysis showed that these S2 neurons receive projections from the contralateral S2 and specifically innervate the layer 4 of primary somatosensory cortex (S1). Microendoscopic calcium imaging and chemogenetic manipulation in freely moving mice showed that this S2 -> S1 feedforward inhibitory circuit mediates ipsilateral pain-induced analgesia, whereas contralateral S2 innervation of the S2 -> S1 circuit mediates contralateral pain-induced analgesia. Our study defines the intra-somatosensory cortical circuits underlying "pain inhibiting pain", expanding the scope of known circuit mechanisms involved in pain relief.

Prosocial behaviors are advantageous to social species, but the neural mechanism(s) through which others receive benefit remain unknown. Here, we found that bystander mice display rescue-like behavior (tongue dragging) toward anesthetized cagemates and found that this tongue dragging promotes arousal from anesthesia through a direct tongue-brain circuit. We found that a direct circuit from the tongue → glutamatergic neurons in the mesencephalic trigeminal nucleus (MTN Glu ) → noradrenergic neurons in the locus coeruleus (LC NE ) drives rapid arousal in the anesthetized mice that receive the rescue-like behavior from bystanders. Artificial inhibition of this circuit abolishes the rapid arousal effect induced by the rescue-like behavior. Further, we revealed that glutamatergic neurons in the paraventricular nucleus of the thalamus (PVT Glu ) that project to the nucleus accumbens shell (NAcSh) mediate the rescue-like behavior. These findings reveal a tongue-brain connection underlying the rapid arousal effects induced by rescue-like behavior and the circuit basis governing this specific form of prosocial behavior.

This study introduces a Fragment-Informed Structure-Activity Relationship (FI-SAR) paradigm for developing cholinesterase inhibitors through strategic coupling of amino-functionalized fragments with quinoline scaffolds. A library of 105 conjugates was synthesized and comprehensively assessed to delineate fragment-to-conjugate activity transfer. Quantitative analysis revealed a positive correlation between fragment potency and conjugate inhibitory activity, with phenolic Mannich base derivatives showing the strongest interdependence. Notably, representative conjugate L4R1-3 achieved sub-nanomolar AChE inhibition (IC₅₀ = 4.6 nM), outperforming clinical references such as donepezil. Furthermore, dual-target inhibitors were constructed (e.g., L1-3, AChE/BChE IC₅₀ = 8.1/0.58 μM) by using FI-SAR strategy. Molecular simulations confirmed stable binding modes of high-activity conjugates within the AChE/BChE active site. This work establishes a streamlined FI-SAR framework, emphasizing that strategic fusion of privileged pharmacophores (e.g., phenolic Mannich bases) with versatile scaffolds like quinoline can accelerate multi-target drug discovery, suggesting broader applicability to enzyme-driven diseases.

Guipi wan (GPW) is a traditional Chinese medicine commonly used in clinical practice, typically to treat neurological diseases such as neurasthenia and traumatic brain injury. It may have positive effects on cerebral ischemia‒reperfusion injury (cI/R). This study aimed to assess the effects of GPW in a mouse model of cI/R and find its possible targets. C57BL/6J mice were used to establish the cI/R model, and the laser speckle doppler was used to determine the success of the model. GPW was administered intragastrically for 7 days, brain tissue sections were stained with TTC, HE, and TUNEL, Western blot assay was performed to detect the effect of apoptosis-related proteins. Furthermore, we screened active ingredients from the TCM Database and constructed a compound‒target network using the Cytoscape 3.8.0 software. Moreover, we employed protein‒protein interaction and component‒target‒pathway network analyses to determine the potential components of GPW and its target genes, the key target was verified through molecular docking. Finally, we detected the influence of the downstream signaling pathway of the target through Western blot. The results showed that GPW decreased the cerebral infarction area, neurological function scores, and neuronal apoptosis in mice by regulating PI3K/AKT signaling pathway. Network analysis indicated that gamma-aminobutyric acid B receptor 1 (GABBR1) might be a potential target for the treatment of cI/R. Molecular docking indicated that 9 active components in GPW could bind to GABBR1 with desirable binding energy. This study represented the demonstratable effect of GPW in the treatment of cI/R injury and suggested GABBR1 as a potential target using network analysis.

Anxiety-associated symptoms following acute stress usually become extinct gradually within a period of time. However, the mechanisms underlying how individuals cope with stress to achieve the extinction of anxiety are not clear. Here we show that acute restraint stress causes an increase in the activity of GABAergic neurons in the CeA (GABA CeA ) in male mice, resulting in anxiety-like behaviors within 12 hours; meanwhile, elevated GABA CeA neuronal CX3CL1 secretion via MST4 (mammalian sterile-20-like kinase 4)-NF-κB-CX3CL1 signaling consequently activates microglia in the CeA. Activated microglia in turn inhibit GABA CeA neuronal activity via the engulfment of their dendritic spines, ultimately leading to the extinction of anxiety-like behaviors induced by restraint stress. These findings reveal a dynamic molecular and cellular mechanism in which microglia drive a negative feedback to inhibit GABA CeA neuronal activity, thus facilitating maintenance of brain homeostasis in response to acute stress.

The newly identified estrogen receptor, G protein-coupled receptor 30 (GPR30), is prevalent in the brain and has been shown to provide significant neuroprotection. Recent studies have linked ferroptosis, a newly characterized form of programmed cell death, closely with cerebral ischemia-reperfusion injury (CIRI), highlighting it as a major contributing factor. Consequently, our research aimed to explore the potential of GPR30 targeting in controlling neuronal ferroptosis and lessening CIRI impacts. Results indicated that GPR30 activation not only improved neurological outcomes and decreased infarct size in a mouse model but also lessened iron accumulation and malondialdehyde formation post-middle cerebral artery occlusion (MCAO). This protective effect extended to increased levels of Nrf2 and GPX4 proteins. Similar protective results were replicated in PC12 cells subjected to Oxygen Glucose Deprivation and Reoxygenation (OGD/R) using the GPR30-specific agonist G1. Importantly, inhibition of Nrf2 with ML385 curtailed the neuroprotective effects of GPR30 activation, suggesting that GPR30 mitigates CIRI primarily through inhibition of neuronal ferroptosis via upregulation of Nrf2 and GPX4.

Aromatic essential oils have been shown to relieve anxiety and enhance relaxation, although the neural circuits underlying these effects have remained unknown. Here, it is found that treatment with 1.0% bergamot essential oil (BEO) exerts anxiolytic-like effects through a neural circuit projecting from the anterior olfactory nucleus (AON) to the anterior cingulate cortex (ACC) in acute restraint stress model mice. Collectively, in vivo two-photon calcium imaging, viral tracing, and whole-cell patch clamp recordings show that inhalation exposure to 1.0% BEO can activate glutamatergic projections from the AON to GABAergic neurons in the ACC, which drives inhibition of local glutamatergic neurons (AONGlu→ACCGABA→Glu). Optogenetic or chemogenetic manipulation of this pathway can recapitulate or abolish the BEO-induced anxiolytic-like behavioral effects in mice with ARS. Beyond depicting a previously unrecognized pathway involved in stress response, this study provides a circuit mechanism for the effects of BEO and suggests a potential target for anxiety treatment.

Narirutin (Nar) is a flavonoid that is abundantly present in citrus fruits and has attracted considerable attention because of its diverse pharmacological activities and low toxicity. Here, we evaluated the preventive effects of Nar in middle cerebral artery occlusion/reperfusion (MCAO/R)-injured mice and oxygen–glucose deprivation/reperfusion (OGD/R)-injured bEnd.3 cells. Pretreatment with Nar (150 mg/kg) for 7 days effectively reduced infarct volume, improved neurological deficits, and significantly inhibited neuronal death in the hippocampus and cortex in MCAO/R-injured mice. Moreover, anti-apoptotic effects of Nar (50 µM) were observed in OGD/R-injured bEnd.3 cells. In addition, Nar pre-administration regulated blood-brain barrier function by increasing tight junction-related protein expression after MCAO/R and OGD/R injury. Nar also inhibited NOD-like receptor protein 3 (NLRP3) inflammasome activation by reducing the expression of thioredoxin-interacting protein (TXNIP) in vivo and in vitro. Taken together, these results provide new evidence for the use of Nar in the prevention and treatment of ischemic stroke.

Chitinase-3-like protein 1 (CHI3L1) is a secreted glycoprotein characterized by its ability to regulate multiple biological processes, such as the inflammatory response and gene transcriptional signaling activation. Abnormal CHI3L1 expression has been associated with multiple neurological disorders and serves as a biomarker for the early detection of several neurodegenerative diseases. Aberrant CHI3L1 expression is also reportedly associated with brain tumor migration and metastasis, as well as contributions to immune escape, playing important roles in brain tumor progression. CHI3L1 is synthesized and secreted mainly by reactive astrocytes in the central nervous system. Thus, targeting astrocytic CHI3L1 could be a promising approach for the treatment of neurological diseases, such as traumatic brain injury, ischemic stroke, Alzheimer's disease, Parkinson's disease, multiple sclerosis, amyotrophic lateral sclerosis, and glioma. Based on current knowledge of CHI3L1, we assume that it acts as a molecule mediating several signaling pathways driving the initiation and progression of neurological disorders. This narrative review is the first to introduce the potential roles of astrocytic CHI3L1 in neurological disorders. We also equally explore astrocytic CHI3L1 mRNA expression under physiological and pathological conditions. Inhibiting CHI3L1 and disrupting its interaction with its receptors through multiple mechanisms of action are briefly discussed. These endeavors highlight the pivotal roles of astrocytic CHI3L1 in neurological disorders and could contribute to the development of effective inhibitors based on the strategy of structure-based drug discovery, which could be an attractive therapeutic approach for neurological disease treatment.

Pain is commonly comorbid with anxiety; however, the neural and molecular mechanisms underlying the comorbid anxiety symptoms in pain (CASP) have not been fully elucidated. In this study, we explored the role of acid-sensing ion channel 1a (ASIC1a), located in GABAergic neurons from the central nucleus of the amygdala (GABACeA), in the regulation of CASP in an acute pain mouse model. We found that the mice displayed significant mechanical pain sensitization and anxiety-like behaviors one day post injection of complete Freud's adjuvant (CFA1D). Electrophysiological recordings from acute brain slices showed that the activity of GABACeA neurons increased in the CFA1D mice compared with that in the saline mice. In addition, chemogenetic inhibition of GABACeA neurons relieved mechanical pain sensitization and anxiety-like behaviors in the CFA1D mice. Interestingly, through pharmacological inhibition and genetic knockdown of ASIC1a in the central nucleus amygdala, we found that downregulation of ASIC1a relieved the hypersensitization of mechanical stimuli and alleviated anxiety-related behaviors, accompanied with reversing the hyperactivity of GABACeA neurons in the CFA 1D mice. In conclusion, our results provide novel insights that ASIC1a in GABACeA neurons regulates anxiety-like behaviors in a mouse model of acute pain.

Diabetic encephalopathy is a common consequence of diabetes mellitus that causes cognitive dysfunction and neuropsychiatric disorders. Praeruptorin C (Pra-C) from the traditional Chinese medicinal herb Peucedanum praeruptorum Dunn. is a potential antioxidant and neuroprotective agent. This study was conducted to investigate the molecular mechanisms underlying the effect of Pra-C on diabetic cognitive impairment. A novel object recognition test and the Morris water maze test were performed to assess the behavioral performance of mice. Electrophysiological recordings were made to monitor synaptic plasticity in the hippocampus. A protein-protein interaction network of putative Pra-C targets was constructed, and molecular docking simulations were performed to predict the potential mechanisms of the action of Pra-C. Protein expression levels were detected by western blotting. Pra-C administration significantly lowered body weight and fasting blood glucose levels and alleviated learning and memory deficits in type 2 diabetic mice. Network pharmacology and molecular docking results suggested that Pra-C affects the PI3K/AKT/GSK3β signaling pathway. Western blot analysis confirmed significant increases in phosphorylated PI3K, AKT, and GSK3β levels in vivo and in vitro upon Pra-C administration. Pra-C alleviated cognitive impairment in type 2 diabetic mice by activating PI3K/AKT/GSK3β pathway.

Hepatocellular carcinoma (HCC) is a common malignant tumor of the digestive system with a low early diagnosis rate. Owing to the side effects, tolerance, and patient contraindications of existing therapies, effective drug treatments for HCC remain a major clinical challenge. However, using approved or investigational drugs not initially intended for cancer therapy is a promising strategy for resolving this problem because their safety have been tested in clinic. Therefore, this study evaluated differentially expressed genes between liver cancer and normal tissues in a cohort of patients with HCC from The Cancer Genome Atlas and applied them to query a connectivity map to identify candidate anti-HCC drugs. As a result, fluphenazine was identified as a candidate for anti-HCC therapy in vitro and in vivo. Fluphenazine suppressed HCC cell proliferation and migration and induced cell cycle arrest and apoptosis, possibly owing to disrupted lysosomal function, blocking autophagy flux. Additionally, in vivo studies demonstrated that fluphenazine suppresses HCC subcutaneous xenografts growth without causing severe side effects. Strikingly, fluphenazine could be used as an analgesic to alleviate oxaliplatin-induced pain as well as pain related anxiety-like behavior. Therefore, fluphenazine could be a novel liver cancer treatment candidate.