OBJECTIVES:To investigate the causal associations between gut microbiota (GM) and Alzheimer's disease (AD) and the mediating role of circulating metabolites using Mendelian randomization (MR) analysis. METHODS:A two-sample MR analysis was conducted based on genome-wide association study (GWAS) summary data. Valid instrumental variables for 473 GM taxa and 233 circulating metabolites were selected, and the inverse-variance weighted (IVW) method was used as the primary analytical approach, with MR-Egger regression and weighted median method as the complementary analyses. Multiple sensitivity analyses were conducted to assess the robustness of the results, and reverse MR analyses were used to verify the direction of causality. Mediation MR analyses were performed to determine the mediating effects of the circulating metabolites. RESULTS:A positive causal association was identified between the abundance of Negativibacillus massiliensis and the risk of AD (OR=1.204, 95% CI: 1.020-1.421,P=0.028), and the results were stable and reliable as confirmed by sensitivity analyses (P>0.05). Reverse MR analysis revealed no significant causal effect of AD on the abundance of Negativibacillus massiliensis (P=0.678). Mediation MR analysis showed that the indirect effect mediated by free cholesterol to total lipid ratio in very small very-low-density lipoprotein (v-VLDL FC/TL) accounted for 6.63% of the total effect of Negativibacillus massiliensis on AD. CONCLUSIONS:From a genetic causal inference perspective, Negativibacillus massiliensis is likely associated with an increased risk of AD, and v-VLDL FC/TL may partially mediate this association, suggesting their potential as targets for AD prevention and treatment.
OBJECTIVES:To investigate outcomes of pregnancy termination in women with pulmonary hypertension (PH) and the correlations of anesthetic management strategies and intraoperative blood pressure variability with postoperative adverse events. METHODS:Sixty-two women underwent termination of pregnancy complicated by PH from June, 2019 to December, 2024 in Guangdong Provincial People's Hospital, including 24 women with mild-to-moderate PH and 38 with severe PH. Perioperative parameters and clinical outcomes of the patients were compared between the two groups. RESULTS:The most common subtype of PH in these women was pulmonary arterial hypertension (82.3%), caused primarily by congenital heart diseases (most frequently atrial or ventricular septal defects). The majority of the women underwent pregnancy termination by dilation and curettage (P=0.02) under general anesthesia (P=0.002). A total of 5 postoperative adverse events were recorded, including PH crisis (3 cases), arrhythmia requiring intervention (1 case), and pulmonary embolism (1 case). After multivariate adjustment, the severity of PH was significantly correlated with both postoperative ICU admission and prolonged postoperative length of stay (P<0.05). Among the patients undergoing dilation and curettage, the incidence of postoperative adverse events was significantly higher in patients receiving intrathecal anesthesia (P=0.014). A high blood pressure variability during surgery was correlated with a high rate of postoperative adverse events (P<0.05), which was significantly lower in patients with a blood pressure variability below the third tertile (OR=10.25, 95% CI=1.06-98.84, P=0.034). CONCLUSIONS:Postoperative recovery following pregnancy termination is associated with the severity of PH. Anesthetic methods for pregnancy termination should be individualized. Patients with a high intraoperative blood pressure variability have an increased risk of postoperative adverse events, and inhaled nitric oxide therapy is effective for reducing pulmonary arterial pressure during surgery.
Neoadjuvant immunotherapy is reshaping the treatment landscape of locally advanced gastric cancer (GC). The central clinical challenge has expanded from whether immunotherapy can enhance treatment efficacy to accurate identification of patients who are most likely to benefit and optimization of the subsequent surgical intervention. Accumulating evidence indicates that perioperative immunotherapy-based regimens can improve pathological complete response (pCR) rates and event-free survival (EFS) of the patients, promoting a shift from conventional chemotherapy plus radical surgery toward biomarker-guided, integrated precision treatment. For therapeutic stratification, microsatellite instability (MSI) status, programmed death-ligand 1 (PD-L1) expression, and refined human epidermal growth factor receptor 2 (HER2) classification have emerged as key indicators for predicting treatment response and selecting eligible patients. Meanwhile, treatment response assessment has evolved beyond conventional radiological evaluation to a multimodal framework incorporating pathological assessment, liquid biopsy, endoscopy, functional imaging, and artificial intelligence (AI)-assisted analysis, thereby providing more comprehensive support for individualized therapeutic decision-making. However, improved treatment efficacy has also raised new questions for surgical management. In particular, discordant responses between primary tumors and regional lymph nodes have challenged whether D2 radical gastrectomy planned according to pretreatment clinical staging remains the optimal surgical strategy. This review systematically summarizes the evolution of neoadjuvant treatment strategies for locally advanced GC, comprehensive response assessment, and redefinition of gastrectomy extent and lymphadenectomy strategy. Looking forward, with the clinical translation of emerging therapeutic targets such as Claudin 18.2 (CLDN18.2) and fibroblast growth factor receptor 2 (FGFR2), together with the maturation of AI-driven multimodal assessment systems, GC surgery may move beyond traditional anatomical boundaries. This transition potentially enables "maximally tolerated radical resection" to "function-preserving surgery guided by biological radicality", ultimately providing truly individualized precision surgical treatment for patients with locally advanced GC.
OBJECTIVES:To investigate the protective effect of lipocalin-2 (LCN2) gene knockout against sepsis-induced acute lung injury (ALI) and vascular endothelial dysfunction in mice and the mediating role of the nuclear factor-κB (NF-κB) signaling pathway. METHODS:Wild-type (WT) C57BL/6 mice receiving sham operation or cecal ligation and puncture (CLP) to induce sepsis were randomized into 3 subgroups for intraperitoneal injections of saline, PDTC, or Bay 11-7082 (n=10). Twenty LCN2 knockout (LCN2KO) mice were randomized for sham operation or CLP modeling. Pulmonary histopathological changes, lung wet-to-dry (W/D) ratio, lung index, Evans blue extravasation, and serum TNF-α and IL-6 levels in the mice were assessed. The colocalization of NF-κB, ICAM-1, and VCAM-1 with CD31 were analyzed using immunofluorescence staining, and endothelial cell apoptosis was examined with TUNEL/CD31 double staining. Western blotting and co-immunoprecipitation assay were performed to analyze the protein expressions of LCN2, p-P65/P65, ICAM-1, VCAM-1 and VE-cadherin and the interaction between LCN2 and P65. RESULTS:The WT mice receiving CLP showed severe lung structural damage with obvious edema and inflammation. In contrast, both the LCN2KO mouse and PDTC-treated WT mouse models of CLP showed milder lung injury with lower W/D ratio, lung index, Evans blue leakage, serum TNF‑α and IL-6 levels and markedly reduced colocalization signals of NF-κB, ICAM-1, and VCAM-1 with CD31. CLP resulted in significantly increased expression levels of LCN2, p-P65/P65, ICAM-1, and VCAM-1, lowered VE-cadherin levels, and enhanced endothelial cell apoptosis, and all these changes were significantly ameliorated in LCN2KO mice and WT mice with NF‑κB inhibitor treatment. Co-immunoprecipitation results suggested the interaction between lung LCN2 and P65. Bay 11-7082 significantly reduced inflammatory cytokine production, downregulated p-P65 expression and its downstream adhesion molecules, and restored VE-cadherin expression. CONCLUSIONS:LCN2 knockout alleviates sepsis-induced ALI and vascular endothelial dysfunction in mice by inhibiting NF-κB signaling.
OBJECTIVES:To investigate the therapeutic mechanism of Chaihu Shugan San (CSS) for treatment of menopausal syndrome (MPS) with liver qi stagnation. METHODS:Fifty-six female C57BL/6 mice were randomized into sham-operated group (n=13) and bilateral ovariectomy (OVX) groups (n=43), and the latter group was further randomized equally into menopausal group, menopausal liver qi stagnation (MPS+CUMS) group subjected to chronic unpredictable mild stress (CUMS), and CSS treatment group (with CSS gavage at daily dose of 6.47 g/kg for 28 days). MPS model establishment was validated by vaginal smears and behavior tests. The levels of sex hormones, neurotransmitters, and inflammatory cytokines in the serum and hippocampus were measured by ELISA. Hippocampal morphology was observed using HE and Golgi staining, and immunofluorescence staining and Western blotting were used to detect the expressions of microglial markers, synaptic plasticity proteins and C3/C3aR pathway proteins in the hippocampal CA1 region. RESULTS:The OVX mice showed disrupted estrous cycles. The MPS+CUMS mice exhibited obvious depression-like behaviors, abnormal sex hormones and neurotransmitters, elevated inflammatory factors, hippocampal CA1 neuronal loss with disordered arrangement and pyknosis, reduced dendritic spines, increased microglial markers, decreased synaptic plasticity proteins, and upregulated C3/C3aR pathway. All these changes were significantly improved or corrected after CSS treatment in MPS+CUMS mice. CONCLUSIONS:CSS alleviates MPS with liver qi stagnation possibly by inhibiting C3/C3aR-mediated M1 microglial polarization and improving synaptic plasticity.
OBJECTIVES:To explore the association between mean arterial pressure variability (MBPV) and in-hospital mortality in patients with non-traumatic subarachnoid hemorrhage (NSAH). METHODS:This study was conducted using the data of NSAH patients from the Medical Information Mart for Intensive Care IV (MIMIC-IV) database and the eICU-CRD, with the final sample size ranging from 1345 to 1820 subjects across different MBPV indicators. Mean arterial pressure (MAP) data within 24, 48, and 72 h after admission were extracted, and the standard deviation (SD), average real variability (ARV), range, and successive variability (SV) of MBPV were calculated. Restricted cubic spline (RCS) combined with Cox regression analysis was adopted to evaluate the nonlinear correlation and determine the risk inflection point using piecewise regression. Time-dependent Cox regression was performed only for variables violating the proportional hazards assumption. Two sensitivity analyses were conducted (excluding patients with surgery within 72 h of admission and those with missing data of medication) to validate the robustness of the results. The primary outcome was all-cause in-hospital mortality. RESULTS:MBPV at all time windows showed a U-shaped nonlinear association with in-hospital mortality of the patients (all P<0.05), and the strongest association was found at 24 h. The inflection point of MBP SD was 8.9, and either excessively low or high fluctuation increased the mortality risk. Time-dependent analysis showed that ARV and SV of MBPV became more hazardous after 4-7 days, while its SD and range were risky mainly within the first 14 days. Both sensitivity analyses confirmed the robustness of the main results. CONCLUSIONS:Early MBPV presents a U-shaped association with in-hospital mortality from NSAH, and moderate fluctuation predicts better prognosis. MBP SDs within 24 and 72 h are the most robust indicators, and the former can serve as the preferred indicator for early blood pressure monitoring.
OBJECTIVES:To investigate the effects of exosomes derived from human umbilical venous endothelial cells (HUVECs) cultured in normoxic and hypoxic conditions (N-exo and H-exo, respectively) on hypoxia-induced injury of human cardiomyocyte AC16 cells and activation of rat cardiac fibroblasts (RCFs) in vitro. METHODS:N-exo and H-exo were extracted from HUVECs by differential centrifugation and sucrose density gradient centrifugation and characterized. AC16 cells and RCFs in normoxic culture or hypoxic culture (1% O₂ for 24 h) were treated with N-exo or H-exo, and the changes in protein and mRNA levels were examined with Western blotting and RT-qPCR. Intracellular ROS levels and apoptosis in AC16 cells were assessed by DCFH-DA staining and TUNEL staining, and the changes in proliferation of RCFs were evaluated with EdU assay. RESULTS:N-exo and H-exo with densities of 1.5×10⁹ and 7.2×10⁸ mL-1 were obtained, respectively, which showed abundant protein components and expressed the exosomal marker HSP70. In AC16 cells, hypoxic exposure significantly upregulated CK-MB mRNA and downregulated Cx43 and GPX4 mRNA levels, causing also enhanced expressions of ALP, IL-1β and cTnT proteins and increased intracellular ROS content and cell apoptosis. These changes were effectively alleviated by treatment with 10⁷ mL-1 N-exo but worsened after treatment with H-exo. In RCFs, hypoxia significantly increased the expressions of collagen I, collagen III and α‑SMA and cell proliferation, which were suppressed by N-exo treatment but further enhanced by H-exo treatment. Hypoxia exposure increased HIF-1α and VEGF expression and phosphorylation level of AKT in both AC16 cells and RCFs; N-exo obviously inhibited while H-exo further enhanced these changes. Px-478, a specific HIF-1α inhibitor, significantly inhibited hypoxia-induced AC16 cell injury and RCF activation. CONCLUSIONS:N-exo derived from HUVECs inhibits while H-exo exacerbates hypoxia-induced AC16 cell injury and RCF activation possibly via the HIF-1α pathway.
OBJECTIVES:To investigate the mechanism of Wuyou Decoction (WYD) for ameliorating hippocampal neuron damage and protecting synaptic plasticity in rats with chronic sleep deprivation (CSD). METHODS:Fifty SD rats were randomly divided into normal control group, CSD model group, and low-, medium-, and high-dose WYD treatment groups (n=10). Except for those in the control group, all the rats were subjected to CSD modeling using the horizontal platform method. Cognitive function of the rats was assessed with Morris water mazetest, and hippocampal pathology, synaptic structure, iron content, oxidative stress markers, and ferroptosis-related protein expressions were evaluated. Molecular docking study was performed to explore the interactions between WYD components and ferroptosis-related proteins. RESULTS:CSD induced significant cognitive impairment and neuronal and synaptic damage in the rat models, causing also increased oxidative stress and iron deposition and upregulated Nrf2 and HO-1 expressions. Treatment with WYD, especially at the medium dose, significantly improved learning and memory abilities of the rat models, alleviated neuronal degeneration, restored synaptic structure, reduced iron accumulation and oxidative stress, and upregulated ferroptosis-related markers as well as Nrf2 and HO-1 expressions. CONCLUSIONS:WYD, optimally at the medium dose, alleviates CSD-induced cognitive impairment in rats by inhibiting ferroptosis, reducing neuronal damage, and enhancing synaptic repair via activation of the Nrf2/HO-1 signaling pathway.
OBJECTIVES:To construct a machine learning-based predictive model for fatty liver in patients with Wilson disease (WD). METHODS:Clinical data retrospectively collected from 1862 WD patients at the First Affiliated Hospital of Anhui University of Chinese Medicine were divided into a training set (70%) and a validation set (30%). The least absolute shrinkage and selection operator (LASSO) was employed to screen the key predictive variables. Seven algorithms, namely logistic regression (LR), decision tree (DT), random forest (RF), extreme gradient boosting (XGBoost), light gradient boosting machine (LightGBM), support vector machine (SVM), and artificial neural network (ANN), were compared for their performance using the area under the receiver-operating characteristic (ROC) curve (AUC), precision-recall (PR) curve, calibration curves, and decision curve analysis (DCA). The contribution of each feature to model prediction was assessed using SHAP analysis. RESULTS:Among the 1862 WD patients, 1296 (69.60%) were complicated with fatty liver. LASSO regression identified platelet count (PLT), red cell distribution width (RDW), alanine aminotransferase (ALT), total bile acids (TBA), type IV collagen (CIV), and indirect bilirubin (IBIL) as the key predictive variables. The LightGBM model demonstrated optimal overall performance, with a training set AUC of 0.826 (95% CI: 0.801-0.849) and good calibration (Brier score 0.143); its validation set AUC was 0.815 (95% CI: 0.776-0.852), and the PR curve showed a high average precision (AP=0.903) with good calibration (Brier score 0.138) and significant clinical net benefit across all the diagnostic thresholds as confirmed by DCA. SHAP analysis indicated that ALT, IBIL, TBA, and CIV all had significant positive effects on model outputs, while RDW and PLT contributed minimally to the cumulative predictive outcomes. CONCLUSIONS:Among the 7 predictive models for fatty liver in WD patients, the LightGBM model demonstrates superior performance to potentially facilitate early screening and risk stratification of WD patients at high risk of fatty liver.
OBJECTIVES:To explore the molecular mechanisms and key pharmacodynamic basis of Baishao for alleviating cancer-related fatigue (CRF) during chemotherapy for breast cancer. METHODS:Bioinformatics analyses were used to explore the active ingredients and potential targets of Baishao, CRF-related targets, and the differentially expressed core genes between chemotherapy and non-chemotherapy groups. Four machine learning algorithms (Random Forest, SVM, XGBoost, and GLM) were used to screen the key feature genes to construct a diagnostic nomogram model with subsequent survival and immunohistochemical analyses using Kaplan-Meier Plotter and HPA databases. In an IL-17-induced Py230 breast cancer cell model of CRF, the regulatory effects of paeoniflorin (a major active ingredient of Baishao) on the core targets were validated using CCK-8 assay, qRT-PCR, Western blotting, and immunofluorescence staining. RESULTS:Thirteen active ingredients (including paeoniflorin, albiflorin, and kaempferol) and 475 drug targets of Baishao were identified, yielding 65 core intersection genes enriched in the MAPK signaling cascade, circadian rhythm, and cell cycle regulation and showing significant correlations with immune cells. The SVM model demonstrated the best diagnostic performance and identified PSMB8, EZH2, CCNE1, PSEN2, and CDK1 as the key feature genes. The SVM-based nomogram achieved a C-index of 0.911, showing excellent calibration and clinical net benefit. Molecular docking confirmed strong binding affinities between the core ingredients of Baishao and the key targets. Survival analysis linked high EZH2 and CCNE1 expressions to poor breast cancer prognosis. In IL-17-induced Py230 cells, chemotherapy resulted in significant upregulation of EZH2 and CCNE1 expressions, which were effectively reversed by paeoniflorin treatment with an efficacy comparable to specific positive inhibitors. CONCLUSIONS:Paeoniflorin alleviates chemotherapy-induced CRF in breast cancer by targeting and inhibiting abnormal expressions of EZH2 and CCNE1 to regulate cell cycle and inflammatory microenvironment, highlighting the therapeutic potential of Baishao for attenuating toxicity and enhancing efficacy of chemotherapy.
OBJECTIVES:To explore the mechanism of Liuwei Dihuang Pill (LWDHW) for improving postmenopausal osteoporosis (PMOP). METHODS:Thirty female C57BL/6 mice were randomized equally into sham-operated group, PMOP model group, and LWDHW treatment group. The mice in the latter two groups underwent ovariectomy to simulate PMOP, followed by gavage of normal saline and LWDHW (twice daily) for 60 days, respectively. Bone mineral density (BMD) of the mice was detected, and TRAP staining was used to observe osteoclast and osteoclast maturation in the tibia tissue. In RANKL-induced RAW264.7 cells with or without lentivirus-mediated CLCF1 knockdown, the effects of incubation with normal rat serum or LWDHW-medicated rat serum on cell membrane fusion rate and F-actin loop formation were observed using immunofluorescence staining with phalloidin peptide; the mRNA and protein expressions of CLCF1, NFATc1, DC-STAMP, OC-STAMP and ATP6V0D2 were detected using RT-qPCR and Western blotting. RESULTS:In PMOP mice, LWDHW treatment significantly increased BMD, reduced osteoclast maturation, osteoclast membrane fusion rate, and F-actin ring formation, upregulated the mRNA and protein expressions of CLCF1 in tibial tissues, and lowered the expressions of NFATc1, DC-STAMP, OC-STAMP and ATP6V0D2. In RANKL-induced RAW264.7 cells with CLCF1 knockdown, treatment with LWDHW-medicated rat serum significantly decreased osteoclast membrane fusion rate, F-actin ring formation, and the expressions of NFATc1, DC-STAMP, OC-STAMP and ATP6V0D2. CONCLUSIONS:LWDHW improves PMOP in mice by regulating osteoclast membrane fusion through upregulating CLCF1 expression and downregulating NFATc1, DC-STAMP, OC-STAMP and ATP6V0D2 expressions.
OBJECTIVES:To investigate the regulatory effect of miR-124 on MK-801-induced anxiety-like behavior in mice. METHODS:Sixty 6-week-old male C57BL/6J mice were randomly divided into control group, MK-801+Saline group, MK-801+Vehicle group, and MK-801+miR-124 group (n=15). Anxiety-like models were established in the latter 3 groups by intraperitoneal injection of MK-801, and stereotaxic brain injection of mmu-miR-124-3p mimics was performed for local miR-124 overexpression in the hippocampus, and successful miR-124 transfection was verified using RT-qPCR. Behavioral tests were conducted to assess the effect of miR-124 overexpression on anxiety-like behaviors of the mice. Western blotting and immunofluorescence staining were used to examine the effects of miR-124 overexpression on hippocampal neuronal protein expressions, neural stem cell number, number of new neurons, and neuronal differentiation. RESULTS:The mice in MK-801+miR-124 group showed significantly increased hippocampal miR-124 levels after the injection. The mice receiving miR-124 microinjection exhibited significant increases in both the total and central zone distance traveled in open field test and in percentages of time spent in the open arms in elevated plus maze test. Hippocampal Tuj1 protein expression and the numbers of hippocampal neural stem cells and newborn neurons, along with the fluorescence area ratio of newborn to mature neurons, increased significantly in the mice in MK-801+miR-124 group. Conclusion miR-124 overexpression ameliorates MK-801-induced anxiety-like behaviors in mice by promoting hippocampal neural stem cell proliferation and differentiation and increasing hippocampal neuron number, suggesting the value of miR-124 as a potential therapeutic target for anxiety disorders.
OBJECTIVES:To investigate palmitoyl-protein thioesterase-1 (PPT1) expression in gastric cancer (GC), its prognostic significance, and its regulatory role in tumor cell lipid metabolism. METHODS:PPT1 expression was analyzed using TCGA-STAD data and immunohistochemistry of clinical samples. In a retrospective cohort of 136 GC patients undergoing radical resection, the association between PPT1 and 5-year survival was evaluated using Kaplan-Meier and Cox regression analyses, and a nomogram was constructed and validated by ROC analysis. GO and REACTOME enrichment analyses were conducted to identify the pathways involved. In cultured GC cell lines with PPT1 knockdown or overexpression, the changes in cell proliferation, migration and invasion were assessed using CCK-8 and Transwell assays. Oil Red O staining, Western blotting and U0126 rescue experiments were performed to explore the role of PPT1-mediated ERK signaling in tumor cell lipid metabolism. RESULTS:PPT1 protein, localized in the cytoplasm, was significantly upregulated in GC tissues compared with adjacent normal tissues. High PPT1 expression was closely correlated with elevated serum CEA and CA19-9 levels and advanced T and N stages (P<0.05), and was an independent risk factor for reduced 5-year postoperative survival. The nomogram incorporating PPT1 showed good predictive accuracy (C-index=0.812) with an AUC of 0.837. In cultured GC cells, PPT1 knockdown significantly suppressed while PPT1 overexpression promoted cell proliferation, migration and invasion. PPT1 overexpression induced abnormal intracellular lipid accumulation in GC cells and significantly upregulated the expressions of the key lipogenic proteins SREBP 1 and FASN, and this pro-lipogenic effect and subsequent ERK signaling activation were effectively reversed by U0126. CONCLUSIONS:PPT1 is overexpressed in GC tissues and predicts poor prognosis. PPT1 overexpression promotes GC progression by activating the ERK signaling pathway and thereby modulating lipid metabolism, suggesting its potential as a novel prognostic biomarker and a therapeutic target for GC.
OBJECTIVES:To construct a high-value region-guided dual-network semi-supervised segmentation method (HVASS) under extremely low labeling rates for addressing the challenges of heterogeneity and blurred boundaries in MRI tumor subregions and improving the accuracy and reliability of complex boundary segmentation for the heart and glioma. METHODS:HVASS employs a dual-network collaborative learning architecture that leverages prediction inconsistency to automatically identify two clinically significant high-risk regions: the high-confidence ambiguity zones, where predictions between the two networks diverge despite at least one network exhibiting high confidence; and the low-confidence stable zones, where predictions agree but with consistently low confidence across both networks. To refine pseudo-label quality, an adaptive dual-teacher mutual distillation mechanism was introduced for dynamically leveraging complementary knowledge from both networks. A high-value region-aware convolutional module was integrated to strengthen feature representation at the tumor margins and the heterogeneous areas, while a wavelet-based frequency-domain refinement module was incorporated to preserve the fine-grained edge details. The framework was evaluated on two publicly available datasets: BraTS2019 for brain glioma MRI and ACDC for cardiac MRI. RESULTS:On the ACDC dataset, HVASS achieved a mean Dice score of 90.34% and a 95th percentile Hausdorff Distance (95HD) of 2.46 mm, representing a 3.24% improvement in Dice and a 2.68 mm reduction in 95HD compared to the state-of-the-art model. On BraTS2019, the model attained a Dice score of 85.07% and a 95HD of 7.68 mm, with a 3.8% increase in Dice for enhancing tumor sub-region and a substantial reduction of boundary localization error. CONCLUSIONS:HVASS demonstrates superior segmentation performance under minimal annotation settings and allows effective capture of fuzzy boundaries and heterogeneous tumor regions in MRI. The method shows particular strength in segmenting small lesions and ill-defined edges and thus lessens the annotation burden of the radiologists.
OBJECTIVES:To investigate whether tetrahydrocurcumin (THC) ameliorates depression-like behaviors in mice by modulating neuroinflammation and neurotrophic balance via the SIRT1/NF‑κB pathway. METHODS:A mouse model of depression was established using chronic restraint stress (CRS). The mice were treated with THC (2.5, 5, or 10 mg/k) or sertraline (10 mg/kg) on a daily basis for 21 consecutive days. Anxiety- and depression-like behaviors of the mice were assessed using open field, elevated plus maze, tail suspension, and forced swimming tests. Serum levels and hippocampal expression levels of SIRT1, p-NF-κB/NF-κB, inflammatory factors (TNF-α and iNOS), and neurotrophic factors (BDNF, GDNF, and TGF-β1) were measured using ELISA, Western blotting, or immunohistochemistry. The expression levels of hippocampal CA3 microglial activation marker (Iba1) and silent information regulator 1 (SIRT1) were analyzed using immunofluorescence staining. In the in vitro experiment, BV2 microglial cells were stimulated with lipopolysaccharide (LPS) and treated with THC with or without the SIRT1 inhibitor EX-527 for validating the pathway mediating the effect of THC. RESULTS:THC treatment significantly alleviated CRS-induced anxiety- and depression-like behaviors and increased body weight gain of the mice. THC obviously reduced serum and hippocampal levels of TNF-α and iNOS, increased the expression levels of SIRT1, BDNF, GDNF, and TGF-β1, and inhibited NF‑κB activation in the hippocampus. The THC-treated mice showed significantly reduced Iba1 expression and Iba1+/SIRT1+ cells and increased SIRT1 expression in the hippocampal CA3 region. In cultured BV2 cells, THC effectively attenuated LPS-induced NF-κB activation, lowered expressions of TNF-α and iNOS, and promoted SIRT1, BDNF, GDNF, and TGF-β1 expressions; these effects were obviously blocked by treatment with EX-527, confirming SIRT1 dependence of the effect of THC. CONCLUSIONS:THC produces significant antidepressant-like effects in CRS mice likely by activating SIRT1 to suppress NF‑κB-mediated neuroinflammation and restore neurotrophic factor expressions. These findings highlight the SIRT1/NF-κB pathway as a key mechanism mediating the therapeutic effect of THC against depression.
OBJECTIVES:To investigate the regulatory effect of everolimus on biological behaviors of rdafitinib-resistant bladder cancer cells and the underlying mechanism. METHODS:An erdafitinib-resistant RT-112-RS cell model was established by continuous induction of RT-112 cells with erdafitinib at the optimized concentration (5 μmol/L) for 16 weeks. The resistant cells were treated with PDGF-BB alone or in combination with 0.01, 0.1, or 1 μmol/L everolimus, and the cytotoxicity (IC50) of everolimus was determined with CCK-8 assay. The changes in cell migration and invasion were evaluated using scratch wound healing assay and Transwell assays. The expressions of the genes of interest in the cells treated with everolimus (0.001- 1 μmol/L) were detected using RT-qPCR, and cell apoptosis was analyzed with flow cytometry. Western blotting was performed to assess phosphorylation levels of the key proteins in the mTORC1/p70S6K, PI3K/Akt, and Raf1/MEK/ERK pathways. RESULTS:Everolimus within the concentration range of 0.001-0.1 μmol/L dose-dependently inhibited RT-112-RS cell proliferation. RT-112-RS cells showed significantly enhanced migration and invasion compared with their parental RT-112 cells. Treatment with 1 μmol/L everolimus significantly suppressed p70S6K phosphorylation but did not result in an enhanced inhibition of cell proliferation effect. Everolimus at 1 μmol/L significantly activated the Raf1/MEK/ERK pathway without affecting PI3K/Akt (Thr308) or mTORC2/Akt (Ser473) phosphorylation. Treatment of the cells with the MEK inhibitor U-0126 obviously reversed everolimus-induced ERK1/2 activation, while the PI3K inhibitor LY294002 only partially reduced ERK1/2 activation in everolimus-treated cells. CONCLUSIONS:Everolimus inhibits RT-112-RS cell proliferation by suppressing p70S6K activity, and at high concentrations, everolimus can result in activation of the PI3K-Raf1/MEK/ERK pathway possibly by relieving negative feedback inhibition to partially offset its inhibitory effect on cell proliferation.
OBJECTIVES:To construct a knowledge graph-based drug repurposing model for predicting potential therapeutic drugs for triple-negative breast cancer (TNBC). METHODS:Drug-target interaction (DTI) affinity data were collected from the BindingDB database and filtered (including data of Kd, Ki, EC50 and IC50). The proposed KGNN model integrates graph convolutional network (GCN)‑extracted drug molecular graph features, ProtBERT-pretrained protein sequence representations, and STRING-derived protein-protein interaction (PPI) knowledge graphs. Multi-head attention mechanisms and gated fusion modules were used to model interaction dependencies. Model performance was evaluated using mean squared error (MSE), Pearson correlation coefficient (PCC), and concordance index (CI). Ablation studies were performed to assess module contributions, and cold-start experiments were conducted to test generalization ability of the model. Using data from TCGA, 1340 TNBC-associated pathogenic genes were screened by bioinformatics analyses and mapped to targets using UniProt. KGNN was applied to predict the candidate drugs, which were validated through molecular docking and molecular dynamics simulations. RESULTS:In the DTI affinity prediction task, KGNN outperformed the benchmark models including KronRLS, SimBoost, DeepDTA, FusionDTA, and GraphDTA (MSE=3.2697, PCC=0.8037, and CI=0.7862). Ablation studies confirmed the critical roles of the modules for enhancing model performance (multi-head attention increased MSE by 5.60%; PPI fusion increased MSE by 10.82%). In cold-start scenarios, KGNN maintained superior performance over the comparators in unseen drug/target settings, demonstrating robust generalization. The TNBC candidate drug predictions well aligned with docking affinities and dynamics simulations (Pearson correlation coefficient>0.85), while attention visualization highlighted the efficacy hotspots. CONCLUSIONS:The KGNN model can effectively predict drug-target interactions to facilitate drug repurposing and the design of multi-target drugs while reducing the screening space and experimental validation costs.
OBJECTIVES:To investigate the effect of electroacupuncture (EA) on chondrocyte apoptosis in rats with knee osteoarthritis (KOA) and the underlying mechanism. METHODS:Twenty-seven SD rats were randomly divided into blank group, KOA model group, and EA group (n=9). Rat models of KOA were established by intra-articular injection of monosodium iodoacetate, and EA treatment was administered at the acupoints "Neixiyan", "Dubi", "Xuehai", and "Yanglingquan" (dense-sparse wave, 2/100 Hz, 0.5-1.0 mA, 20 min/day for 14 days). Knee function of the rats was assessed using Lequesne MG scale, and cartilage pathologies and ultrastructural changes were observed with HE staining and transmission electron microscopy. The protein expressions of TLR4, MyD88, NF-κB p65, COL2, COL10, Bax, Bcl-2, and caspase-3 and mRNA levels of TLR4, MyD88, and NF‑κB p65 were detected using Western blotting and RT-qPCR; serum levels of IL-1β and TNF‑α were determined with ELISA. Another 15 rats were randomized into blank control, model, TLR4 inhibitor (TAK-242), EA, and sham-EA groups (n=3) for analysis of HMGB1 and key pathway protein expressions and NF‑κB p65 nuclear translocation. RESULTS:The KOA rats showed increased Lequesne MG scores, obvious chondrocyte injuries, upregulated TLR4, MyD88, NF‑κB p65, COL10, Bax, and caspase-3 expressions, increased serum IL-1β and TNF‑α levels with decreased COL2 and Bcl-2 expressions. EA significantly reduced Lequesne MG scores, improved cartilage pathologies and reversed the abnormal changes in protein expressions and inflammatory cytokines. The KOA rats showed increased HMGB1 expression and enhanced NF‑κB p65 nuclear translocation, and EA treatment effectively decreased HMGB1 expression, reduced nuclear NF‑κB p65, and increased cytoplasmic NF‑κB p65. The TLR4 inhibitor produced similar effects, while sham EA produced no significant effect. CONCLUSIONS:EA alleviates cartilage injury in KOA rats possibly by inhibiting HMGB1-mediated activation of TLR4/MyD88/NF‑κB signaling and NF‑κB p65 nuclear translocation to regulate Bcl-2/Bax balance and suppress mitochondria-dependent chondrocyte apoptosis.
OBJECTIVES:To investigate the inhibitory effect of Escherichia coli outer membrane vesicles (E. coli-OMVs) on glioma proliferation and the underlying mechanism. METHODS:CCK-8 assay and EdU incorporation assay were used to determine the optimal concentrations of E. coli-OMVs and chlorpromazine (CPZ) for cell treatment. C6 glioma cells treated with 10 μg/mL E. coli-OMVs and 10 μmol/L CPZ were examined for PCNA and Ki67 expressions and cell cycle distribution using qRT-PCR, Western blotting, immunocytochemistry and flow cytometry. The expressions of β-catenin, cyclin D1, and c-Myc proteins in C6 cells were detected following treatment with OMVs in the presence or absence of CHIR99021. In a SD rat model bearing orthotopic glioma, the effects of intranasal PBS or OMVs (0.25, 0.5, 1.0 mg/kg) administration on tumor growth and PCNA and Ki67 expressions were observed using fluorescence imaging and immunohistochemistry. RESULTS:E. coli-OMVs within the concentrations of 2.5-10 μg/mL dose-dependently inhibited C6 cell viability, and such inhibitory effect was attenuated by 2.5, 5, and 10 μmol/L CPZ, which was the most effective at 10 μmol/L. The OMVs at 10 μg/mL significantly reduced EdU-positive cells, downregulated mRNA and protein expressions of PCNA and Ki67, and increased G0/G1-phase and decreased S-phase cell populations. These alterations were largely reversed by 10 μmol/L CPZ. Treatment with OMVs downregulated cellular expressions of β‑catenin, Cyclin D1, and c-Myc, which were partially restored by application of CHIR99021. In the tumor-bearing mice, intranasal OMVs administration dose-dependently decreased the relative fluorescence signals, slowed tumor growth, and reduced PCNA and Ki67 expressions. CONCLUSIONS:E. coli-OMVs inhibit glioma cell proliferation and delay orthotopic tumor growth in rats likely via endocytic uptake of the OMVs to cause suppression of Wnt/β-catenin signaling and cell cycle arrest.
OBJECTIVES:To investigate the effects of electroacupuncture (EA) on cognitive function and neuroinflammation in a rat model of vascular dementia (VD) and the underlying mechanism. METHODS:Sixty male SD rats were randomly assigned to sham-operated group (n=10) and VD model group (n=50) receiving bilateral common carotid artery occlusion. Thirty rats with successful VD modeling were randomized into model group, EA group, and donepezil treatment group (n=10). EA treatment was administered at the acupoints Baihui (GV20) and Shenting (GV24) with a disperse-dense wave (2/15 Hz, 1 mA, 30 min/day), and donepezil was given by gavage at 0.45 mg/kg. Both interventions lasted 28 days. Cognitive function of the rats was assessed using Morris water maze test, and neuronal pathologies were observed using HE and Nissl staining. GFAP-labeled astrocyte activation was assessed by immunohistochemistry, and astrocytic ultrastructure was examined with transmission electron microscopy. GFAP/p-NF-κB colocalization was detected by immunofluorescence staining. Hippocampal IL-1β, IL-6, and TNF-α levels were measured by ELISA, and the protein expression levels of C3, S100A10, TLR4, and MyD88 and the p-NF-κB/NF‑κB ratio were detected by Western blotting. RESULTS:Compared with the sham-operated rats, VD rats showed significant cognitive impairment, obvious neuronal disorganization and pyknosis in the hippocampus, excessive astrocyte activation, increased GFAP/p-NF‑κB colocalization, inflammatory cytokine levels and expressions of C3 and TLR4/MyD88/NF-κB pathway proteins, and decreased expression of S100A10. Treatment with EA and donepezil significantly improved the performance of the rats in Morris water maze test, alleviated neuronal injury, inhibited astrocyte overactivation and ultrastructural damage, reduced inflammatory cytokine levels, expressions of C3, TLR4, and MyD88 proteins and the p-NF-κB/NF-κB ratio, and increased the expression of S100A10 in the hippocampus. CONCLUSIONS:EA at GV20 and GV24 improves cognitive impairment and attenuate neuroinflammation in VD rats possibly by inhibiting TLR4/MyD88/NF-κB signaling and regulating astrocytic A1/A2-like phenotypic imbalance.
