Search Results (264)

Lemur tail kinase 3 (LMTK3) is an oncogenic Ser/Thr kinase implicated in breast cancer (BC) progression, therapy resistance, and poor clinical outcomes, yet the molecular mechanisms governing its regulation remain poorly understood, particularly the role of its C-terminal intrinsically disordered region (IDR). Given that IDRs frequently harbour hidden structural motifs that control protein dynamics, we combined computational, biophysical, and biochemical approaches to systematically map regulatory elements within the LMTK3 C-terminus, identifying two regions (residues 688–1095 and 1181–1486) that interact with the LMTK3 kinase domain (LMTK3-KD). Characterisation of these interactions revealed that LMTK3^1181–1486 displays preferential binding to inactive wild-type LMTK3-KD over a constitutively active mutant (LMTK3-KD^L313R), a behaviour consistent with a potential autoinhibitory interaction. Guided by AlphaFold3 modelling, we localised this interaction primarily to a short α-helical motif (α-helix 2; residues 1247–1258) within the C-terminal IDR and subsequently identified Ser1258 within this motif as a candidate regulatory phosphorylation site, using [γ-³²P]-ATP kinase assays and mass spectrometry. Phosphorylation at Ser1258 altered interactions between α-helix 2 and the kinase domain, reducing binding to wild-type LMTK3-KD while increasing affinity for LMTK3-KD^L313R. Functionally, phospho-null mutation of Ser1258 impaired oestrogen receptor alpha (ERα) upregulation, proliferation, migration, and clonogenicity in ER-positive BC cell lines, and reduced tumour growth in female BALB/c nude mice bearing orthotopic MCF7 xenografts. Together, these findings identify a previously uncharacterised regulatory element within the LMTK3 C-terminus and support a model in which Ser1258 phosphorylation modulates kinase domain interactions and LMTK3-driven oncogenic functions in BC. Full article

Malpighia mexicana has been traditionally used in Mexican ethnomedicine for the treatment of central nervous systems disorders. Previous studies have demonstrated neuropharmacological activity of crude extracts, but the contribution of enriched phytochemical fractions remains unclear. A phenolic-enriched fraction (G4) obtained from the aerial parts of M. mexicana was evaluated for antidepressant-like and anticonvulsant activities in mice. Antidepressant-like effects were assessed using the forced swimming test (FST) and tail suspension test (TST), while locomotor activity was evaluated in the open field test. Anticonvulsant activity was determined using the pentylentetrazole (PTZ)-induced seizure model. The phytochemical profile of G4 was characterized by HPLC-DAD analysis. G4 (50 and 200 mg/kg, p.o.) significantly reduced immobility time in both FST and TST compared to the vehicle group (p < 0.05) without affecting locomotor activity in the OFT, suggesting a specific antidepressant-like effect. In the PTZ model, G4 significantly increased latency to clonic and tonic seizures and reduced seizure severity. HPLC-DAD analysis revealed the presence of phenolic compounds, with kaempferol and p-coumaric acid as major constituents. The phenolic-enriched G4 fraction from M. mexicana exhibits antidepressant-like and anticonvulsant effects in murine models, supporting its traditional use. These findings suggest that phenolic constituents, particularly kaempferol and p-coumaric acid, may contribute to the observed neuropharmacological activity. However, since only the G4 fraction was pharmacologically evaluated and not the isolated compounds, the specific role of each constituent remains to be established. Further studies are required to elucidate the underlying mechanisms of action and the contribution of individual compounds and potential synergistic interactions. Full article

Background: Secondary lymphedema is a chronic complication of oncologic surgery and radiotherapy for which effective disease-modifying therapies remain lacking. While mesenchymal stem cells (MSCs) have shown partial benefit in experimental models, direct functional comparison with lineage-committed lymphatic endothelial progenitor cells (LEPCs) remains limited. Methods: Secondary lymphedema was induced in C57BL/6J mice by circumferential tail skin excision with disruption of superficial lymphatics. Animals received intradermal phosphate-buffered saline (PBS), MSCs, or adipose-derived LEPCs on postoperative days 1 and 7. Lymphatic function was longitudinally quantified using IVIS-based near-infrared indocyanine green (ICG) imaging with standardized region-of-interest analysis. Tail diameter was measured serially throughout follow-up as a complementary morphometric parameter. Tissue remodeling was assessed by Picrosirius Red staining and qualitative LYVE-1 immunofluorescence. Statistical analysis incorporated mixed-effects modeling to evaluate treatment group, sex, time, and their interaction terms, with estimation of effect sizes and 95% confidence intervals. Results: LEPC-treated mice demonstrated higher IVIS signal intensity than MSC- and PBS-treated animals, consistent with improved lymphatic transport at the injury site. Longitudinal tail diameter analysis showed a more favorable temporal profile in the LEPC group relative to controls, with exploratory analysis suggesting potential variations in temporal profiles between sexes that warrant further investigation in larger cohorts. Histologic analysis showed reduced non-tissue/void area and a more favorable qualitative pattern of LYVE-1 staining in LEPC-treated tissue. Conclusions: LEPC administration enhanced functional and structural recovery in a murine model of secondary lymphedema. These findings support further translational evaluation of LEPC-based approaches as a regenerative strategy for secondary lymphedema. Full article

Background/Objectives: Atopic dermatitis (AD) is a multifactorial inflammatory skin disorder in which epidermal barrier disruption and dysregulated immune responses drive persistent cutaneous inflammation. Owing to their broad spectrum of biological activities, marine-derived peptides have attracted increasing attention as potential therapeutic agents capable of modulating inflammatory and immune pathways. Methods: In this study, a novel peptide, WPN-15 (NGAIADQQPQRPNIV), was isolated from enzymatic hydrolysates of walleye pollock (Gadus chalcogrammus) tail by-products using an activity-guided purification process consisting of dialysis, fast protein liquid chromatography-gel permeation chromatography (FPLC-GPC), reverse-phase high-performance liquid chromatography (RP-HPLC), and electrospray ionization mass spectrometry (ESI-MS). The anti-inflammatory activity of WPN-15 was first examined in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages, and subsequently validated in a 2,4-dinitrochlorobenzene (DNCB)-induced atopic dermatitis model using six-week-old male BALB/c mice. Results: WPN-15 significantly inhibited nitric oxide production in LPS-stimulated macrophages without causing cytotoxic effects. Topical administration of WPN-15 markedly alleviated DNCB-induced AD-like kin lesions, significantly reduced dermatitis severity scores, and decreased serum interleukin (IL)-6 levels. Histological evaluation further demonstrated that WPN-15 attenuated epidermal hyperplasia, dermal thickening, and mast cell infiltration. Furthermore, WPN-15 significantly downregulated the mRNA expression of IL-1β and IL-6 and inhibited signal transducer and activator of transcription 3 (STAT3) phosphorylation in skin tissues, indicating that its protective effects are mediated, at least in part, through the suppression of the IL-6/STAT3 signaling pathway. Conclusions: WPN-15 effectively attenuated inflammatory responses and pathological features associated with experimental AD. These findings demonstrate that walleye pollock tail by-products represent a valuable and sustainable source of bioactive peptides and support the potential application of WPN-15 as a marine-derived therapeutic candidate for the management of AD. Full article

Salvia guaranitica A.St.-Hil. Ex Benth. (Lamiaceae) is native from South America, traditionally used for releasing tension and treating gastrointestinal cramps. It was evaluated for the presence of flavonoids and isoflavones in the extract, and the hypothesis of being cardioprotective under ischemia and reperfusion (I/R), antidiarrheal, antispasmodic and anxyolitic in respective models. For phytochemistry, GC-MS and HPLC-DAD methodologies were used. Cardiac performance after subchronic oral administration of S. guaranitica tincture (S.g-T, 0.95 mg extract/mL) was evaluated in rat isolated perfused hearts exposed to I/R, and mechanisms were characterized. Antispasmodic effects of S.g-T were evaluated on contractile concentration–response curves (CRCs) of carbachol (CCh) and calcium (Ca²⁺) in rat isolated intestinal tissue. In vivo tests of ricin-oil diarrhea models, open-field (OFT), novel-suppressed feeding (NFT) and tail suspension (TST) tests were performed on mice. Phenolic acids, genistin, luteolin, kaempferol and quercetin glycosides, genins, loliolide, dodecane, farnesene, phytol and isochiapin B were identified. Post-ischemic cardiac recovery was improved by S.g.-T, associated to activation of phosphatidylinositol-3-kinase and β-estrogenic receptor. S.g.-T also induced antispasmodic effects, as a non-competitive inhibitor of the CCh-CRC and Ca²⁺-CRC in the intestine. In vivo, S.g.-T showed antidiarrheal activity (95 mg extract/kg) and anxiolytic-like effects (0.95–9.5 mg extract/kg) without sedation or antidepressant-like effects. Therefore, S. guaranitica leaves have potential therapeutic cardioprotective, antidiarrheal, antispasmodic and anxiolytic effects. Full article

Major depressive disorder (MDD) involves dysregulation of the hypothalamic–pituitary–adrenal axis, neuroinflammation, serotonergic dysfunction, and impaired neurotrophic signaling. Whether combining corticosterone (CORT) with chronic restraint stress (CRS) produces a more comprehensive depression-like phenotype than either model alone remains unexplored. Male C57BL/6N mice were assigned to four groups—Sham (n = 7), CORT (20 mg/kg s.c.; n = 7), CRS (3 h/day; n = 7), and CORT + CRS (C+C; n = 8)—and evaluated by behavioral tests, hippocampal qRT-PCR, Western blot, LC-MS/MS metabolomics, and immunohistochemistry. All experimental groups showed elevated immobility in tail suspension and forced swim tests without inter-group differences. Two-way ANOVA revealed a behavioral–molecular dissociation: FST immobility showed CORT and CRS main effects without a significant interaction, whereas hippocampal LC-MS/MS analytes exhibited strong CORT × CRS interactions. The C+C group showed the strongest 5-HT1A receptor (HTR1A) upregulation, the greatest reductions in Trk-b and DCX, and unique decreases in serum dopamine, glutamine, and adenosine. Representative Western blot densitometry indicated the largest p-ERK/ERK reduction in C+C (54%), while p-CREB/CREB was most suppressed in CRS (52%). BDNF and NeuN proteins were lowest in C+C (68% and 61% reductions). Combined CORT + CRS produces the most comprehensive depression-related molecular profile, integrating multiple pathological domains. Full article

Background: Depression-associated memory impairment is a clinically significant comorbidity that responds poorly to conventional antidepressants. Dihydromyricetin (DHM), a natural dihydroflavonoid, exhibits neuroprotective and antidepressant-like effects, but its impact on stress-induced memory deficits and the underlying mechanisms remain unclear. Methods: A chronic restraint stress (CRS) mouse model was employed. DHM was intraperitoneally administered daily (20 mg/kg bodyweight) for 8 consecutive days. Depression-like behavior and memory functions were assessed with the tail suspension test, the object recognition memory (ORM) task, and the Y-maze test. Network pharmacology, molecular docking, and molecular dynamics (MD) simulations were integrated to predict potential targets and associated signal pathways. Expression of hippocampal PKA, p-CREB, CREB, and BDNF was measured through Western blotting. Results: CRS significantly increased immobility time in the tail suspension test and impaired both recognition memory (ORM) and spatial working memory (Y-maze), without affecting locomotor activity. DHM treatment effectively reversed behavioral despair and memory deficits. Network pharmacology identified 47 intersecting targets between DHM and depression-related memory impairment, with CREB1, ESR1 and NOS1 as core hub nodes. KEGG enrichment particularly implicated the involvement of the cAMP signaling pathway and other signaling pathways, which are associated with depression and/or memory impairment. Molecular docking predicted strong binding affinities of DHM to CREB1 (−9.225 kcal/mol), NOS1 (−9.113 kcal/mol) and ESR1 (−7.903 kcal/mol). MD simulations confirmed the thermodynamic stability of DHM-ESR1 and DHM-NOS1 complexes. Western blot analysis demonstrated that DHM treatment significantly increased the levels of hippocampal PKA, p-CREB and BDNF, without altering total CREB expression in CRS mice. Conclusions: DHM alleviated CRS-induced depression-like behaviors and memory deficits, possibly through regulation of multiple signaling pathways, including the hippocampal PKA-CREB-BDNF signaling pathway. These findings highlight DHM as a promising natural therapeutic candidate for depression-associated cognitive impairment. Full article

Quantifying the spatial distribution of immune cells within intact skin tissue is essential for understanding diseases such as vitiligo, in which CD8⁺ T cells selectively destroy epidermal melanocytes within the discrete, parallelogram-shaped epidermal compartments of mouse tail skin, which we term scales. Existing workflows rely on manual region drawing, which is labor-intensive and operator-dependent. Here we present ScaleNet, a three-stage deep-learning pipeline for automated per-scale quantification of whole-mount immunofluorescent images, implemented as an Imaris XTension to enable seamless integration with existing 3D imaging workflows. ScaleNet (i) encodes a 3D confocal volume as a pseudo-RGB projection that preserves height information lost by standard maximum-intensity projection, (ii) applies two independently trained Detectron2 Mask R-CNN models—one for epidermal scales and one for hair follicles—with sliced inference (SAHI) to segment whole-mount images at full resolution, and (iii) maps the resulting 2D mask back into the Imaris 3D coordinate system to quantify user-defined Spot objects per scale. Applied to vitiligo mice imaging, ScaleNet produced per-scale counts of CD8⁺ T cells and DCT⁺ melanocytes, enabling unbiased spatial statistics in the tail epidermis, demonstrating that ScaleNet can provide the quantitative spatial resolution needed to dissect the micro-anatomical dynamics of autoimmune depigmentation. Full article

The tryptophan–kynurenine pathway, mediated by indoleamine 2,3-dioxygenase 1 (IDO1) and tryptophan 2,3-dioxygenase (TDO), is critically involved in the pathogenesis of depression. A series of novel morpholine–coumarin derivatives were designed and synthesized as dual IDO1/TDO inhibitors. Through in vitro enzyme screening, compound 14d exhibited potent inhibitory activity with IC₅₀ values of 0.34 µM and 0.75 µM, respectively. In lipopolysaccharide (LPS)-stimulated BV2 microglial cells, 14d downregulated IDO1/TDO expression, suppressed pro-inflammatory cytokines (IL-1β, COX-2, iNOS, TNF-α), and upregulated the anti-inflammatory cytokine IL-10. In an LPS-induced acute depressive mouse model established in C57BL/6 mice, intraperitoneal administration of 14d (20 mg/kg) significantly reduced immobility time in the forced swim and tail suspension tests, without affecting spontaneous locomotor activity. Mechanistic studies revealed that 14d inhibited microglial activation in the hippocampal dentate gyrus, reduced cerebral kynurenine levels, increased serotonin content, and upregulated BDNF/PKA signaling. Molecular docking further predicted the binding interactions of 14d with the active sites of IDO1 and TDO. These findings suggest that 14d represents a promising dual IDO1/TDO inhibitor lead compound for the treatment of inflammation-associated depression through modulation of the kynurenine pathway and neuroinflammatory responses. Full article

Essential oils are multicomponent natural products with potential neurobehavioral activity, but integrated comparative studies remain limited. This study compared the essential oils of Satureja brevicalyx, Peperomia dolabriformis, and Rosmarinus officinalis in relation to their chemical profiles, predicted target interactions, preliminary acute oral safety, anxiolytic-like and antidepressant-like effects, antagonist-sensitive behavioral patterns, and exploratory serum biomarkers. Oils were characterized by GC-MS, and their constituents were screened by molecular docking against anxiety-, depression-, sleep-, and stress-related targets. Independent cohorts of male BALB/c mice received oral essential oils (25–100 mg/kg) and were assessed in anxiety-related, depression-related, and locomotor behavioral paradigms, including the elevated plus maze, light–dark box, marble burying, tail suspension, forced swim, and open field tests. Flumazenil and WAY-100635 were used to examine whether the behavioral responses were sensitive to γ-aminobutyric acid type A (GABA-A)/benzodiazepine- and serotonin 1A (5-HT1A)-related pharmacological modulation, respectively. In a preliminary 24-h acute oral toxicity screen, no mortality was observed up to 5000 mg/kg. The three oils produced anxiolytic-like and antidepressant-like effects without reducing spontaneous locomotor activity. Within its experimental block, S. brevicalyx showed the most consistent flumazenil-sensitive anxiolytic-like pattern and FDR-significant reductions in corticosterone and TNF-α, together with increased IL-4. P. dolabriformis showed a broader predicted multitarget docking profile and antagonist-sensitive behavioral attenuation compatible with mixed pathway participation. R. officinalis produced significant but more moderate behavioral effects. WAY-100635 partially attenuated the antidepressant-like effects of all three oils. These findings support differentiated but convergent functional neurobehavioral profiles among the oils. The docking, antagonist, and biomarker results should be interpreted as hypothesis-generating evidence of possible pathway involvement, supporting further validation in chronic stress models, receptor-specific assays, pharmacokinetic studies, and expanded safety evaluations. Full article

Background: Schizophrenia is a chronic neuropsychiatric disorder characterized by cognitive impairment, behavioral abnormalities, neuroinflammation, and oxidative stress. Increasing evidence suggests that dysregulated inflammatory cytokines and impaired antioxidant defenses contribute to schizophrenia pathophysiology. This study investigated the neuroprotective and anti-inflammatory effects of Gallic Acid (GA) in a ketamine-induced experimental schizophrenia model. Methods: Thirty male Balb/C mice were randomly divided into control, ketamine, and ketamine + GA groups. Schizophrenia was induced with ketamine (25 mg/kg/day) for 7 days, while the treatment group additionally received GA (60 mg/kg/day) for another 7 days. Behavioral tests, including open field, novel object recognition, and tail suspension tests, were performed to evaluate locomotor activity, cognition, and depressive-like behavior. Tumor necrosis factor-alpha (TNF-α), interleukin-1 beta (IL-1β), interleukin-18 (IL-18), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px), total antioxidant status (TAS), and total oxidant status (TOS) levels were analyzed in hippocampal and prefrontal cortex tissues to assess inflammatory and oxidative stress-related alterations. Results: Ketamine induced schizophrenia-like behaviors, including hyperlocomotion, memory impairment, and increased immobility. These behavioral alterations were accompanied by significantly elevated TNF-α, IL-1β, IL-18, and TOS levels, alongside reduced SOD, CAT, GSH-Px, and TAS levels in the hippocampus and prefrontal cortex. GA treatment ameliorated behavioral impairments, restored antioxidant enzymes, increased TAS levels, and reduced pro-inflammatory cytokines and TOS in these brain regions. Conclusions: GA exerted neuroprotective effects in the ketamine-induced schizophrenia model by reducing oxidative stress, neuroinflammation, and behavioral deficits. These findings suggest that Gallic Acid may serve as a promising therapeutic candidate for schizophrenia through modulation of inflammatory and oxidative stress pathways. Full article

Hulunbuir short-tailed sheep (HSTS) and Hu sheep (HS) exhibit distinct tail phenotypes linked to ecological adaptation, with HSTS carrying a loss-of-function mutation (c.G334T) in the T gene while HS retain the wild-type allele. However, the cellular and molecular mechanisms underlying T-mediated tail development remain unclear. Here, we performed single-cell RNA sequencing on HSTS and HS embryos at embryonic days 16 and 19 (E16 and E19), complemented by cross-species validation using a CRISPR/Cas9 mouse model carrying the same mutation. We identified 12 cell types in E16 HSTS and E16 HS embryos, and 15 cell types in E19 HSTS and E19 HS embryos and found that the MDK_ITGA6+ITGB1 ligand–receptor pair consistently mediated core intercellular communication. The MDK_ITGA6+ITGB1 axis mediates intercellular communication critical for tail bud formation; BMP activation and FGF repression disrupt AER survival, leading to tail shortening. Developmental trajectories showed a shift from early progenitor states at E16 to terminal differentiation at E19. Crucially, HSTS embryos showed transcriptomic signatures consistent with premature AER regression. The T mutation showed transcriptomic signatures of increased BMP pathway activity and reduced FGF8 expression, which may disrupt AER survival and contribute to the short-tail phenotype. In the mouse model, mutant T expression was reduced, and expression dynamics of WNT5B and FGF8 were perturbed, corroborating the sheep findings; however, homozygous T mutation causes embryonic lethality in mice but not in sheep, indicating species-specific differences. This study provides single-cell transcriptomic evidence linking the T c.G334T mutation to premature AER regression in sheep, complemented by cross-species validation in a CRISPR/Cas9 mouse model, offering new insights into the cellular mechanisms of tail development and may provide a basis for future investigations into tail-related breeding markers, pending experimental validation. These changes are associated with AER maintenance and tail outgrowth. Full article

Objective: In this study, Dehydroepiandrosterone (DHEA-induced Polycystic Ovary Syndrome (PCOS) mice were used as models to evaluate the improvement effect of Vitexin (Vit) on ovarian fibrosis and explore the mechanism of action of the NR4A1/NLRP3 signaling pathway. Method: Sixty 4-week-old female ICR mice of the same batch number were selected and their systems were divided into 6 groups (n = 10): normal (Control, Ctrl) group, model (Polycystic Ovary Syndrome, PCOS) group, treatment (Vitexin, The Vit group, normal NR4A1 gene silencing group (Ctrl NR4A1^-/-), NR4A1 gene silencing model group (PCOS NR4A1^-/-), and NR4A1 gene silencing treatment group (Vit NR4A1^-/-). Silencing gene modeling was performed by tail vein injection of adeno-associated virus (serotype AAV-8), and the mouse genotypes were detected by qRT-PCR technology 14 days after injection. After the genotype was determined, the PCOS group and the PCOS NR4A1^-/- group were administered dehydroepandrosterone (6 mg/100 g/d) by gavage for 28 consecutive days for modeling, while the Vit group and the Vit NR4A1^-/- group were treated with dehydroepandrosterone + vitexin (10 mg/kg/d) by gavage for 28 consecutive days. All mice were raised with pure water and regular maintenance food. After 4 weeks of drug intervention, the mice were euthanized and samples were collected. The pathological changes in ovarian tissue were observed by H&E staining, and the degree of ovarian tissue fibrosis was observed by Masson staining. The levels of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px), malondialdehyde (MDA), total cholesterol (TC), triglycerides (TG), high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C) in mouse serum were detected by biochemical kits. The levels of inflammatory factors (IL-1β, IL-6, IL-18, TNF-α) in mouse serum were determined by enzyme-linked immunosorbent assay. Real-time fluorescence quantitative PCR (qRT-PCR) was used to detect oxidative kinase (Gsta4, Prdx3, Mgst1, Gpx3, Gsr), inflammatory factors (Nlrp3, Caspase-1, Asc, Il-1β, Il-18, Tnf-α) and fibrotic pathway-related genes (Tgf-β1, Smad3, Collagen1, CTGF, α-SMA, Mmp-13, and β-catenin) in ovarian tissues. The levels of inflammatory factors (NLRP3, Caspase-1, ASC, IL-1β, IL-18, TNF-α, IκBα) and fibrosis in mice were determined by Western blot method, and statistical description and analysis were performed using SPSS software. Result: In the wild-type genotype group, compared with the PCOS group, Vit treatment could effectively regulate the metabolic abnormalities of PCOS mice, including inhibiting excessive weight gain, restoring normal glucose tolerance, and reducing body fat content. After Vit treatment, the levels of MDA, TC, TG, LDL, IL-1β, IL-6, IL-18 and TNF-α in the serum of PCOS mice were significantly reduced, while the levels of SOD and HDL in the serum of PCOS mice were increased. The staining results indicated that Vit treatment could significantly inhibit the process of ovarian fibrosis in PCOS mice. The results of WB and PCR demonstrated that after Vit gavage treatment in mice, inflammatory and fibrotic factors such as Nlrp3, Caspase-1, Asc, Il-1β, Il-18, Tgf-β1, Smad3, Collagen1, CTGF, and α-SMA in ovarian tissues could be significantly down-regulated, and the fibrotic level of ovarian tissues could be reduced. Among the same measurement indicators, the silenced NR4A1 group showed a certain degree of increase compared with the wild genotype group, but there was no significant difference. Conclusions: Vit intervention can restore the sex hormone levels and follicular development in ovarian tissues of PCOS mice, regulate reproductive endocrine disorders and abnormal lipid metabolism levels, and regulate the expression of Collagen I, a-SMA and CTGF in the ovaries by inhibiting the NR4A1/NLRP3 signaling pathway, thereby improving the ovarian fibrosis level of PCOS mice. It is suggested that it may play a key role in the treatment of PCOS and the prevention and delay of its long-term complications. Full article

Background: In some children with immunodeficiency, Bacille Calmette–Guérin (BCG) vaccination can lead to dissemination and severe infection, including severe intracranial infection, called disseminated BCG disease (BCGosis), which is characterized by high rates of disability and mortality. However, the specific routes by which BCG crosses CNS barriers and the patterns of temporal remodeling of the CNS immune microenvironment during infection have yet to be fully elucidated. Methods: Mice were infected with BCG through tail vein injection to construct an intracerebral mycobacterial infection mouse model, wherein the brain was collected and analyzed using single-cell RNA sequencing. We profiled temporal transcriptomic changes in cell populations, pathways, and cell–cell communication associated with anti-mycobacterial activity and inflammation-induced disturbance of physiological brain activities. Results: After BCG was injected via tail vein, histopathological images and cultured colonies of brain tissue confirmed successful brain infection. Then, whole-brain tissue was dissected for 10× Genomics single-cell sequencing, and we acquired 15 cell types. Dysfunction and inflammatory responses were observed in endothelial and ependymal cells. Infection induced dynamic state transitions in microglia, enabling their differentiation into disease-related and interferon-responsive states. Along with peripheral immune cells, microglia formed temporally structured communication networks that mediated early events such as chemokine recruitment and inflammatory storms, and facilitated late-stage immune checkpoint upregulation. Conclusions: This study proposes BCSFB as a possible pathway of mycobacteria invasion and reveals the temporality of immune response processes in the pathogenesis of intracerebral mycobacterial infection. Full article

Salivary adenoid cystic carcinoma (SACC) is a malignant salivary gland neoplasm characterized by aggressive local invasion and a marked propensity for metastasis. However, the role of MEOX1 in SACC progression remains poorly defined. In this study, we examined the effects of MEOX1 overexpression on the malignant behavior of SACC cells in vitro and in vivo. Human SACC-83 and SACC-LM cells were transduced with lentiviral vectors encoding MEOX1 or an empty vector control, and cell proliferation, migration, invasion, and cell cycle distribution were assessed using CCK-8, wound healing, Transwell, and flow cytometric assays, respectively. RNA sequencing was performed to characterize transcriptional changes associated with MEOX1 overexpression. In vivo, tumor growth was evaluated in BALB/c nude mice bearing subcutaneous xenografts, and pulmonary metastatic colonization was assessed using a tail vein injection model. MEOX1 overexpression reduced the proliferation, migration, and invasion of SACC cells in vitro and increased the G2/M phase fraction. In xenograft models, MEOX1-overexpressing cells formed smaller tumors and showed lower Ki67 staining than control cells. In the experimental lung metastasis model, mice injected with MEOX1-overexpressing cells developed fewer pulmonary metastatic nodules. RNA-seq identified 588 differentially expressed genes associated with MEOX1 overexpression, with enrichment in pathways including cytokine–cytokine receptor interaction, Toll-like receptor signaling, and G protein-coupled receptor signaling. Together, these findings indicate that enforced MEOX1 expression is associated with reduced malignant phenotypes in SACC models and with transcriptomic alterations in pathways related to immune response, G protein-coupled receptor signaling, and DNA damage response. Full article