Neuroinflammatory and immune-mediated processes are increasingly recognized as important contributors to adult ischemic stroke pathobiology, contributing not only to acute neuronal injury and secondary tissue damage, but also to later phases of repair and recovery. Experimental and early clinical evidence suggests that immunotherapeutic interventions may modulate post-ischemic inflammatory cascades and immune-cell activation, thereby potentially contributing to neuroprotective and neurorestorative responses. However, the clinical efficacy, safety, and translational relevance of these approaches remain incompletely defined. This systematic review aimed to critically synthesize contemporary preclinical and clinical evidence on neuroinflammation-targeted immunotherapeutic strategies in adult ischemic stroke, while distinguishing direct immunotherapy from regenerative cell-based approaches with immunomodulatory relevance. A systematic search of PubMed/MEDLINE and Embase was conducted in October 2025 and updated in January 2026 to identify original English-language studies published between January 2020 and December 2025. Eligible studies included preclinical and clinical investigations evaluating direct immunotherapeutic interventions or regenerative cell-based therapies with immunomodulatory relevance in adult ischemic stroke, compared with placebo, standard care, or control conditions appropriate to study design. In total, 913 records were screened, of which 55 studies met all inclusion criteria and were included in the qualitative synthesis. Of these, three randomized controlled trials (N = 637) provided comparatively compatible, although methodologically heterogeneous, outcome data for an exploratory meta-analysis of excellent functional outcome at day 90. Methodological quality was assessed using validated design-specific tools for randomized controlled trials, observational studies, and preclinical animal experiments. Primary outcomes included functional recovery and infarct-related measures, while secondary outcomes encompassed neuroinflammatory biomarkers, immune modulation, and safety. Therapeutic approaches were categorized into molecular immunotherapy, biological immunotherapy, and regenerative cell-based therapies with immunomodulatory properties, according to their dominant mechanism of action and translational rationale. Molecular interventions targeting inflammatory and immunometabolic signaling pathways were associated with reductions in neuroinflammatory signaling and infarct-related measures in preclinical models in preclinical models, although corresponding human evidence remained limited. Regenerative cell-based therapies, including mesenchymal stromal/stem cells, progenitor cells, and related cellular products, demonstrated neuroregenerative, paracrine, and immunomodulatory characteristics in preclinical and early translational studies with favorable safety profiles; however, clinical functional benefits remained modest, heterogeneous, and inconsistent. Biological approaches, particularly monoclonal antibodies targeting leukocyte adhesion and immune-cell trafficking, demonstrated generally acceptable safety profiles but did not demonstrate consistent functional benefit across currently available randomized clinical trials. In the exploratory quantitative synthesis, no significant pooled clinical benefit was observed (OR = 0.87; 95% CI, 0.27-2.77), with moderate between-study heterogeneity (I2 = 46.3%). Cross-study comparability was further limited by substantial heterogeneity in intervention timing, dosing regimens, patient selection, biological targets, and outcome definitions, precluding definitive conclusions regarding efficacy. Neuroinflammation-targeted immunotherapy in adult ischemic stroke, together with selected regenerative cell-based strategies of immunomodulatory relevance, appears biologically plausible and mechanistically supported by current preclinical and early clinical evidence based on current preclinical and early clinical evidence and has demonstrated generally acceptable safety profiles in early-phase studies; however, current evidence remains insufficient to support consistent clinical benefit, and available trials may be insufficiently powered to reliably detect moderate but clinically meaningful treatment effects. Future research should prioritize mechanism-driven trial designs, standardized outcome measures, biomarker-informed patient stratification, optimized therapeutic windows, and integrative strategies combining immunomodulation with established reperfusion therapies. Well-powered, methodologically harmonized clinical trials aligned with the temporal and biological heterogeneity of post-stroke inflammation are essential to clarify translational potential and define the role of immune-targeted therapies in stroke management. Importantly, the present review supports a mechanistically differentiated framework in which direct immunotherapy and regenerative cell-based therapy should not be treated as interchangeable categories, even when both modulate post-stroke neuroinflammation. Overall, the available evidence may support further investigation of a stage-specific and mechanistically differentiated model of immune-targeted intervention in ischemic stroke.
Berberine, curcumin, biochanin A, cucurbitacin E, and caffeic acid phenethyl ester (CAPE) are plant-derived compounds with long histories of use in traditional medicine for inflammatory and proliferative conditions. Their known capacity to modulate NF-κB signaling makes them candidates for anticancer investigation, particularly in mesenchymal malignancies such as fibrosarcoma, which arise in muscle-rich environments shared with normal myogenic tissue. To evaluate the selective anticancer potential of these compounds in fibrosarcoma (WEHI-164) and normal muscle (L6) cells, with focus on mitochondrial function, mitophagy, cellular senescence, and NF-κB-related metabolic pathways, alongside preliminary in vivo toxicity assessment. IC50 values were determined using MTT and PrestoBlue® assays. Mitochondrial membrane potential was assessed using JC-1 and normalized to the matched untreated control for each cell line, and mitophagy by PINK1/PARKIN immunofluorescence colocalisation together with a mitophagy dye assay. Cellular senescence was measured using a β-galactosidase assay, and ATP levels by a luminescence-based method. Gene expression of NF-κB pathway components and PFKFB3 was analyzed by RT-qPCR. In vivo-like toxicity was assessed using the Galleria mellonella model, including PBS handling, DMSO vehicle, and 70% ethanol utility controls, with survival data analyzed by Kaplan-Meier curves and the log-rank test. The compounds differentially affected normal and cancer cells, indicating selectivity toward malignant phenotypes. Decreased ATP and mitochondrial depolarization suggest disruption of bioenergetic homeostasis, supported by modulation of mitophagy. Stronger effects in WEHI-164 cells indicate higher susceptibility to mitochondrial dysfunction. Increased cellular senescence suggests inhibition of tumor proliferation. These findings indicate that natural NF-κB modulators may exert anticancer effects by targeting mitochondrial and metabolic homeostasis. Differential sensitivity between normal and tumor cells highlights therapeutic potential. In the G. mellonella model, berberine and curcumin did not differ significantly from the PBS or DMSO controls, whereas CAPE, CurE, and particularly biochanin A produced significantly greater larval mortality. The G. mellonella assay should be regarded only as a preliminary acute toxicity screen, and further in vivo studies in mammalian models are required to clarify mechanisms and clinical relevance.
Clostridium perfringens (C. perfringens) disease in Bactrian camels is an acute infectious disease with high mortality, causing serious economic losses to the breeding industry. However, studies on its epidemiological patterns and whole-genome characteristics remain limited. This study aimed to investigate the epidemiological patterns and whole-genome characteristics of C. perfringens disease in Bactrian camels. Epidemiological investigations, physiological parameter measurements, pathological analyses, isolation and identification of C. perfringens, and whole-genome sequencing were performed to elucidate the epidemiological patterns and whole-genome characteristics of C. perfringens disease in Bactrian camels. The incidence of C. perfringens disease in Bactrian camels was 37.89% (72/190), with a mortality rate of 54.17% (39/72). Compared with healthy camels, affected camels exhibited elevated body temperature, respiration, and pulse, along with significantly increased hemoglobin levels and red cell distribution width (p < 0.01). Elevated alkaline phosphatase, amylase, and phosphorus levels were also observed (P < 0.01). Autopsy revealed hemorrhagic gastroenteritis; enlargement of the liver, kidneys, spleen, and other organs; and cerebral congestion and edema. Histopathological examination revealed edema, hemorrhage, and congestion in parenchymal organs, along with reduced neuronal numbers and neuronal shrinkage in brain tissue. Three toxin genes, plc, etx and cpe, were detected by polymerase chain reaction. Whole-genome sequencing analysis revealed that the total genome length of the eight strains of C. perfringens ranged from 3, 195, 592 bp to 3, 595, 833 bp, and the predicted number of coding genes ranged from 2, 897 to 3, 413. Gene Ontology database annotation indicated enrichment in metabolic processes, cellular processes, catalytic activity, binding, and cellular component categories. Kyoto Encyclopedia of Genes and Genomes annotation identified pathways related to cellular community-prokaryotes, signaling molecules and interactions, translation, and metabolism. Virulence Factors of Pathogenic Bacteria Database annotation identified virulence factors, including alpha-toxin, C. perfringens enterotoxin, theta-toxin, alpha-clostripain, kappa-toxin, mu-toxin, and sialidase. Six drug resistance genes (vanh, cpir, vant, vanw, vany and mPRF) were annotated by the Comprehensive Antibiotic Resistance Database. In addition, three novel sequence types (ST1057, ST1058, and ST1059) were identified. Phylogenetic analysis based on the core genome revealed that the strains isolated from Bactrian camel showed substantial genomic divergence relative to strains isolated from other animals. In young Bactrian camels, C. perfringens is predominantly associated with type A and D strains, which exhibit distinct genomic characteristics, providing a foundation for improved diagnosis, prevention, and vaccine development.
Chronic obstructive pulmonary disease (COPD) is a progressive respiratory disorder characterized by persistent airflow limitation and chronic airway inflammation. Current therapeutic strategies primarily offer symptomatic relief and are often limited by systemic side effects, inadequate lung deposition, and poor patient compliance. Naringin (NAR), a natural flavonoid with strong antioxidant, anti-inflammatory, and anti-fibrotic activities, has demonstrated potential in mitigating COPD-associated pathophysiology. However, its therapeutic application is restricted by poor water solubility, low bioavailability, and rapid metabolism. Nanotechnology-based drug delivery systems, particularly poly(lactic-co-glycolic acid) (PLGA) nanoparticles, provide an effective approach for lung-targeted therapy. Their nanoscale size promotes deep lung deposition, enhanced cellular uptake, reduced lung clearance, improved therapeutic efficacy, and reduced systemic side effects. The present study aimed to develop NAR-loaded PLGA nanoparticles (NAR PLGA NP) for enhanced cell-targeting in inflammatory lung conditions. NAR PLGA NP were prepared using the emulsion solvent evaporation method, with PLGA in the organic phase and soya lecithin (SL) with poly(vinyl alcohol) (PVA) as surfactants in the aqueous phase. A face-centered central composite design was employed to optimize the formulation. The optimized nanoparticles were characterized for size distribution by dynamic light scattering, entrapment efficiency, Transmission Electron Microscopy (TEM), Fourier Transform Infrared (FTIR), Differential Scanning Calorimetry (DSC), X-Ray Diffraction (XRD), and in vitro drug release. The safety of PLGA and lecithin-coated PLGA nanoparticles (LC PLGA NP) was assessed using an MTT assay on lung epithelial cells, followed by cellular uptake studies, angiogenesis by chick Yolk Sac Membrane (YSM) assay, and in vitro evaluation of reactive oxidative stress (ROS) and anti-inflammatory activity. The optimized PLGA formulation showed a hydrodynamic diameter of 201 ± 1 nm with PDI 0.20 ± 0.03 and EE of 76.11 ± 2.1%, and 81.7 ± 4.9% drug release at 72 h, whereas LC PLGA NP showed a hydrodynamic diameter of 308 ± 3 nm, PDI of 0.21 ± 0.05, entrapment efficiency of 82.45 ± 4.8%, and 71.4 ± 3.2% drug release at 72 h. Both PLGA NP and LC PLGA NP demonstrated good cytocompatibility with lung epithelial cells, efficient cellular uptake, and a significant reduction in intracellular reactive oxygen species (ROS) levels (**** p value < 0.0001). Moreover, the formulations markedly suppressed pro-inflammatory cytokines, including TNF-α, IL-6, and IL-1β, indicating anti-inflammatory activity. The angiogenesis assay further suggested their ability for lung tissue repair and remodeling. These findings support the potential of LC PLGA NP as a promising cell-specific targeting system for naringin in inflammatory lung conditions.
Urotensin-II is a powerful vasoconstrictor peptide. Both the peptide and its receptor, GPR14, affect central nervous system, cardiovascular system, and kidney functions. Urotensin-II was found to be involved in the development of many solid organ tumors. The peptide and its receptor have been demonstrated to be present in breast cancer tissue. Moreover, its human plasma level is increased in breast cancer patients. It has also been reported that some cancer patients may have urotensin gene polymorphisms. Antagonists of this peptide receptor with different chemical structures have been produced and studied in different disease models. There are no experimental or clinical studies investigating the effects of urotensin-II receptor antagonists on breast cancer. In this study, MCF-7 cells were grown in an appropriate environment and then plated in wells. WST-8 was used to detect cell proliferation and inhibition effects. The expression levels of the MTA-1, ESR1, Kiss1, UT-II and UTR genes were measured via quantitative RT‒PCR. This study is the first to show the effects of urotensin-II peptide and its receptor antagonists on breast cancer. While urotensin-II increased cell proliferation, all the antagonists had inhibitory effects. When the expression of genes affecting the metastasis or suppression of tumor cells was investigated, urotensin-II increased MTA-1 and ESR1 expression and decreased Kiss1 expression. Urotensin-II receptor antagonists reversed these effects. As a result, while urotensin-II triggered breast cancer cell proliferation, its receptor antagonists had inhibitory effects on proliferation and positive regulatory effects on tumor suppressor genes. These results suggest that urotensin-II receptor antagonists may provide effective results for breast cancer treatment.
Cisplatin is a key treatment for head and neck squamous cell carcinoma (HNSCC), but the development of resistance severely limits its effectiveness. The molecular determinants underlying cisplatin resistance in HNSCC remain unclear. Multiple databases were used to screen the core genes related to cisplatin resistance in HNSCC patients. Tumor tissue samples from HNSCC patients were collected and the expression of FOXA2 was verified through various pathological tests to establish the correlation between FOXA2 expression and the clinical characteristics of the patients. The in vitro and patient-derived organoids (PDOs) models were used to verify the regulatory effect of FOXA2 on the cisplatin resistance of HNSCC. Transcriptome sequencing combined with multi-omics analysis demonstrated that LAMC2 is a downstream target of FOXA2 in regulating cisplatin resistance. Bioinformatic screening of cisplatin-resistant cohorts revealed that FOXA2 was the only gene significantly associated with poor survival outcomes in TCGA-HNSCC patients. Transcriptomic profiling and pathway enrichment analyses revealed the activation of the PI3K/AKT signaling cascade. We identified LAMC2 as a direct transcriptional target of FOXA2. Chromatin immunoprecipitation and luciferase reporter assays confirmed FOXA2 binding to the LAMC2 promoter, resulting in transcriptional activation. FOXA2-mediated upregulation of LAMC2 increased PI3K and AKT phosphorylation, and LAMC2 overexpression reversed the impaired malignant phenotypes caused by FOXA2 silencing. In xenograft models and PDO systems, FOXA2 overexpression reduced responsiveness to cisplatin, whereas FOXA2 inhibition significantly increased therapeutic sensitivity. Our research has identified a previously unrecognized regulatory axis involving FOXA2, LAMC2, and PI3K/AKT, which plays a crucial role in the progression and resistance to cisplatin in HNSCC. Therefore, targeting the FOXA2-LAMC2 axis may represent a novel therapeutic strategy to overcome cisplatin resistance in HNSCC.
Paraformaldehyde-fixed mouse liver samples are sectioned on a vibratome at 100 µm. The free-floating sections are permeabilized, followed by exposure to one of two heat-induced antigen retrieval methods selected based on the antigen of interest. Antigen retrieval is carried out in a mechanical stabilization apparatus developed here to prevent free-floating sections from deforming. The immunostaining is then carried out with noted modifications to previously established protocols. Spatial organ biology widely relies on slicing tissues into one cell-thick sections. These studies have advanced our understanding of organ structure and function; yet, thin sections have been insufficient to investigate three-dimensional organization in complex organs. A more recent approach utilizes thicker sections, enabling visualization of volumetric tissue architecture. Currently, few protocols exist to strengthen immunofluorescence signal on thick tissue sections while maintaining tissue integrity, particularly for difficult antigens. Here, we developed a simple, affordable method that mechanically stabilizes thick tissue sections and preserves tissue integrity during heat-mediated antigen retrieval used to enhance antigen accessibility. This method widens the utility of thick tissue sections and facilitates holistic analysis of complex tissues, opening new possibilities for spatial organ research.
Gouty arthritis (GA) is an inflammatory joint disease caused by the deposition of monosodium urate (MSU) crystals within the joint space and surrounding tissues. In traditional Chinese medicine, Rhizoma Drynariae (Gusuibu) has long been widely used in the clinical treatment of GA, and flavonoids are considered its key bioactive constituents. This research employed network pharmacology to construct a component-target network of total flavonoids of Rhizoma Drynariae (TFRD) against GA, thereby identifying key components, core targets, and related pathways. Rat models were established by intra-articular injection of a monosodium urate crystal suspension and treated with TFRD or the positive control, colchicine, by oral gavage. After sample collection, network pharmacology-based prediction results were subsequently validated using rat serum metabolomics, enzyme-linked immunosorbent assay (ELISA), and Western blot analysis. Network pharmacology analysis indicated that the anti-GA effects of TFRD are mediated through key targets, including IL6, AKT1, TNF, EGFR, JUN, and PTGS2, and are mainly associated with inflammation, immune, and apoptosis-related pathways, such as the IL-17, TNF, NF-κB, MAPK, PI3K-AKT, JAK-STAT, and T-cell receptor signaling pathways. Similarly, metabolomics also uncovered the pivotal roles of the inflammatory response. Hematoxylin and eosin (H&E) staining confirmed that TFRD reduced infiltration of inflammatory cells. ELISA assay confirmed that the TFRD group significantly inhibited the expression of inflammatory factors TNF-α, IL-6, and IL-17A in synovial tissue. Western blot analysis revealed that TFRD inhibited the GA-induced hyperphosphorylation of AKT, MAPK p38, and NF-κB p65 in rat synovial tissue. TFRD can effectively ameliorate the inflammation-triggered changes in the GA rats by directly modulating related inflammatory factors and pathways.
The postoperative wound-healing response significantly affects the outcome of glaucoma filtering surgery. Understanding this response at the cellular level is crucial. This study investigated the effects of the Rho kinase inhibitor ripasudil on immune cells activated by monocyte chemoattractant protein-1 (MCP-1), a known risk factor for glaucoma filtering surgery failure. Single-cell RNA sequencing and immunocytochemical analyses were performed to identify the type of lysozyme M (LysM)-positive cells. LysM-eGFP knock-in mice were observed using multiphoton microscopy after MCP-1 loading. Raw image data were analyzed with Imaris software to quantify the number and velocity of LysM-positive cells. In the Boyden chamber assay, THP-1 and RAW264.7 cells were loaded into the upper chambers, with MCP-1 in the lower wells acting as a chemoattractant. The effect of ripasudil was examined in each experiment. LysM-positive cells were identified as macrophages in mouse subconjunctival tissue. MCP-1 significantly increased both the velocity and number of LysM-positive cells compared with the intact state. Pretreatment with ripasudil significantly reduced cell velocity from 6.48 ± 3.87 to 2.97 ± 1.59 µm/min (P < 0.0001, Welch's t-test), although the change in cell number was not significant. The Boyden chamber assay showed that MCP-1 induced THP-1 and RAW264.7 cell migration, which was almost completely inhibited by 10 µM ripasudil. Ripasudil attenuated MCP-1-activated macrophage behavior in both intravital imaging and in vitro experiments. These findings suggest that ripasudil may have therapeutic potential for preventing excessive wound healing after glaucoma filtering surgery.
Porcine astrovirus 4 (PoAstV4) is an emerging pathogen that has previously been associated with respiratory disease in piglets. The objective of this study was to inoculate naïve piglets with PoAstV4 and characterize the host-pathogen interaction and pathological lesions. Cesarean-derived colostrum-deprived (CDCD) piglets were inoculated intratracheally and intranasally with PoAstV4 PCR-positive tissue homogenate previously screened for the presence of other primary pathogens with next-generation sequencing (NGS). Nasal and fecal swabs for PCR identification of PoAstV4 were collected pre and postinoculation. Animals were euthanized at 5, 8, and 21 days postinoculation (DPI) and tissues were collected for histopathologic and digital image analysis. Nasal swabs were PCR positive for PoAstV4 from 2 DPI to 10 DPI. Microscopic lesions of epitheliotropic viral infection of the trachea and bronchi, including epithelial attenuation and lymphoplasmacytic infiltration, were found in inoculated pigs at 5 DPI and 8 DPI. PoAstV4 was localized to the lesions demonstrated by in situ hybridization and quantified by digital image analysis. In addition, PoAstV4-inoculated pigs at 8 DPI had an abundance of infiltrating lymphocytes predominated by T cell lymphocytes characterized by immunohistochemistry (IHC) and quantified using digital image analysis. Both anti-PoAstV4 IgM and IgG were detected in serum, with IgG levels first detectable at 14 DPI and increasing to the end of the study at 21 DPI. The presented findings confirm the respiratory epithelial tropism of PoAstV4 while also characterizing viral shedding and host immune response dynamics.
To identify in situ stages of entosis in non-small cell lung cancer (NSCLC) tissue and to assess the impact of neoadjuvant chemotherapy (NACT) on their frequency. The study included 9 patients with morphologically verified NSCLC (stage T1-2N0-1M0-1) classified according to the WHO classification (2021). Four patients received NACT (vinorelbine + carboplatin), while five underwent surgery without prior therapy. Histological specimens were examined using Whole Slide Imaging (Pannoramic Mirax Midi, Carl Zeiss, Germany) and analyzed with Panoramic Viewer software. Entotic events were identified according to Mackay's criteria. Statistical analysis was performed using the Mann-Whitney U test. All five lysosomal stages of entosis were successfully identified in situ. Stage II was the most frequently observed. In patients treated with NACT, Stage III was more common, suggesting a slowing of this phase, while the frequency of Stage V was significantly lower (p=0.01). A trend toward an overall increase in entotic events after NACT was noted; however, when normalized to tumor section area, no significant differences between groups were detected. For the first time in situ, variants of entomammoptosis were identified, including binucleated "loser" cells and mitosis of the internalized cell. Neoadjuvant chemotherapy does not increase the overall frequency of entosis in NSCLC tissue but alters the distribution of stages, prolonging Stage III and reducing the completion of the process at Stage V. The results expand current understanding of the role of entosis in tumor cell adaptation and the development of drug resistance in NSCLC. Энтоз представляет собой форму программируемой клеточной гибели, характеризующуюся внедрением одной клетки в другую. Большинство данных об энтозе получено на клеточных культурах, тогда как исследования in situ в опухолевой ткани ограничены. Выявить in situ стадии энтоза в ткани немелкоклеточного рака легкого (НМРЛ) и оценить влияние неоадъювантной химиотерапии (НАХТ) на их частоту. В исследование включено 9 пациентов с НМРЛ стадии T1—2N0—1M0—1, подтвержденным морфологически в соответствии с классификацией ВОЗ (2021). При этом 4 пациента получили НАХТ (винорельбин + карбоплатин), 5 оперированы без предшествующей терапии. Гистологические препараты исследовали с использованием Whole Slide Imaging (Pannoramic Mirax Midi, Carl Zeiss, Германия) и программы Panoramic Viewer. Энтотические события идентифицировали по критериям Маккея. Статистическую обработку проводили с применением критерия Манна—Уитни. Удалось идентифицировать все 5 стадий лизосомального энтоза in situ. Наиболее часто фиксировалась II стадия. У пациентов после НАХТ чаще наблюдалась III стадия, что указывает на замедление этого этапа, тогда как частота V стадии была достоверно ниже (p=0,01). На уровне тенденции отмечено повышение суммарного числа энтотических событий после НАХТ, однако при нормировании на площадь среза различий между группами не выявлено. Впервые in situ обнаружены варианты энтомаммоптоза — двухъядерные клетки-«лузеры» и митоз внутренней клетки. Предоперационная химиотерапия не увеличивает общую частоту энтоза в ткани НМРЛ, но изменяет распределение стадий, замедляя III стадию и снижая завершенность процесса на V стадии. Эти данные позволяют предположить существование паразитического варианта энтоза, при котором внутренняя клетка избегает действия химиопрепаратов.
Ischemia-percussion (I/R) injury describes a paradoxical phenomenon in which blood flow restoration following ischemia not only fails to salvage the tissue but also exacerbates tissue injury. Due to the unique physiological functions, cell types, and metabolic requirements of different tissues, their responses and vulnerability to ischemia/reperfusion injuries vary significantly. The limited understanding of complex mechanisms underlying I/R injury in different organs further thwarts the advancement of effective therapeutic interventions, the translational application of promising interventional strategies still faces many obstacles. Salidroside, a natural bioactive constituent of traditional medicinal plants, demonstrates great potential in combating I/R injuries in various organs. Salidroside exerts organ-specific regulatory effects based on the unique characteristics of damage in different organs, simultaneously alleviates I/R injuries by common mechanism include inhibiting oxidative stress, reducing inflammatory responses, regulating cell apoptosis, and inhibiting ferroptosis. To summarize the evidence, we searched the PubMed and Web of Science databases from inception to February 28, 2026, using "salidroside" (and its synonym "rhodioloside") and "ischemia-reperfusion injury" (and its MeSH term "reperfusion injury") as core keywords. This narrative review summarizes the common and organ-specific protective mechanisms of salidroside against I/R injury across different organs and points out the limitations of existing studies in terms of evidence quality, dose standardization, thereby providing research directions for further development of related pharmaceutical formulations and their clinical translation.
Capecitabine (CAP), a chemotherapy prodrug, is associated with a significant dose-limiting hepatotoxicity. This study evaluated the protective efficacy of melatonin (MEL) and cerium oxide nanoparticles (CeO₂ NPs) against CAP-induced toxicity. Fifty male Wistar rats were divided into five groups: control, CAP-only (500 mg/kg/week), CAP + MEL (10 mg/kg/day), CAP + CeO₂ NPs (30 mg/kg/day), and a combination therapy group (CAP + MEL + CeO₂ NPs). After a six-week treatment period, assessments included alanine and aspartate transaminases (ALT, AST); hepatic oxidative stress markers [glutathione peroxidase-1 (GPx-1), superoxide dismutase (SOD), and malondialdehyde (MDA)]; apoptotic markers [B-cell lymphoma 2 (Bcl-2), Bcl-2-associated X protein (BAX) mRNA, cleaved caspase-3 (c-caspase-3), and protein 53 (p53)]; pro-survival signaling [extracellular signal-regulated kinase (ERK1/2)]; and histopathological evaluation of liver tissue. CAP administration induced significant liver damage, characterized by elevated ALT and AST, depletion of GPx-1 and SOD, increased MDA, and activation of the mitochondrial apoptotic pathway, indicated by an elevated BAX/Bcl-2 ratio and increased levels of p53 and c-caspase-3. This was accompanied by suppression of ERK1/2 signaling and severe histopathological alterations. Both MEL and CeO₂ NPs, significantly attenuated these deleterious effects. The combination therapy demonstrated a comparatively superior effect, resulting in a near-complete normalization of all measured parameters, including oxidative stress, apoptosis, ERK1/2 activity, and liver histoarchitecture. Melatonin and CeO₂ NPs provide hepatoprotection via distinct yet complementary mechanisms. Their combined administration presents a superior strategy to mitigate CAP-induced liver injury by concurrently counteracting oxidative stress, inhibiting apoptosis, and enhancing pro-survival signaling.
Left atrial remodeling (LAR) critically contributes to the progression of heart failure (HF) and the development of atrial fibrillation (AF) following myocardial infarction (MI). The protein α2δ1, primarily known for its role in neuropathic pain, is abundantly expressed in atrial tissue, but its involvement in post-MI LAR remains unclear. Here, LAR models were established in rats post-MI, and atrial hypertrophy was induced in HL-1 cells using angiotensin II (AngII). The role of α2δ1 in atrial hypertrophy was examined through treatment with either the α2δ1 inhibitor gabapentin or a C-terminal interfering peptide (α2δ1 CT-pep). A significant upregulation of α2δ1 expression was observed in the left atrium (LA) of MI rats and in AngII-treated HL-1 cells. Western blot analysis revealed increased α2δ1 levels in membrane fractions and decreased levels in the cytoplasmic fractions compared to controls. Both gabapentin and α2δ1 CT-pep treatment significantly reduced HL-1 cell hypertrophy and inhibited CAMKII and HDAC4 phosphorylation. Co-immunoprecipitation assays demonstrated an interaction between α2δ1 and GluN1, which was enhanced by AngII stimulation. Inhibition of α2δ1 attenuated the α2δ1-GluN1 interaction and reduced GluN1 translocation to the plasma membrane. In MI-induced HF rats, gabapentin treatment diminished atrial hypertrophy, suppressed AF inducibility and duration, and decreased membrane-associated α2δ1 and GluN1 levels. These findings suggest that the C-terminal domain of α2δ1 may contribute to left atrial hypertrophy in chronic ischemic heart failure and is associated with altered membrane GluN1 abundance and p-CAMKII/p-HDAC4 signaling. α2δ1 may therefore represent a potential therapeutic target for left atrial remodeling in ischemic heart failure.
Relapsed/metastatic anaplastic thyroid carcinoma (R/M ATC) is associated with poor prognosis. Approximately 40% of ATCs harbor the BRAFV600E mutation, for which targeted therapy is available. However, the clinical characteristics of BRAFV600E-mutant versus wild-type ATC and the role of liquid biopsy (LB) for molecular assessment and monitoring remain incompletely defined. Consecutive patients with R/M ATCs treated at a tertiary referral center were stratified as BRAF wild-type (Cohort A) or BRAFV600E-mutant (Cohort B). Patients received chemotherapy, dabrafenib and trametinib (BRAF/MEKi), immune checkpoint inhibitors (ICI) or best supportive care. Tumor tissue (TB) was analyzed by RT-PCR and/or next-generation sequencing (NGS). Circulating cell-free DNA was assessed by droplet digital PCR (ddPCR) and/or a 52-gene NGS panel on LB at baseline in both cohorts and longitudinally in Cohort B. Clinical characteristics, treatment outcomes, and concordance between TB and LB were evaluated. Between 2018 and 2021, 24 patients were included (13 Cohort A, 11 Cohort B). BRAFV600E tumors more frequently presented with metastatic disease at diagnosis (81.8% vs. 15.4%, p = 0.003). Overall, 79% received first-line therapy and 41.6% second-line treatment. All patients in Cohort B received BRAF/MEK inhibitors, achieving a median progression-free survival of 3.2 months (95% CI 1.2-17.5). Concordance for BRAFV600E status between TB and LB assessed by ddPCR was 93.7% (specificity 100%, sensitivity 85.7%). Among 12 paired TB/LB NGS analyses, overall mutation concordance was 30%. In longitudinal assessment, LB anticipated disease progression in 3 of 6 cases and, in one case, identified emerging secondary alterations during BRAF/MEK inhibitor therapy. BRAFV600E-mutant ATC displays distinct clinical features and improved survival when treated with targeted therapy; ddPCR-based liquid biopsy provides a rapid and sensitive method for BRAFV600E detection and may support timely therapeutic decision-making. Serial LB analysis may contribute to disease monitoring and detection of resistance mechanisms in selected patients.
Microglia are the resident immune cells of the human central nervous system and play key roles in development, homeostasis, and disease. These functions are mediated by a broad repertoire of cell-surface receptors, including G protein-coupled receptors such as the ADP receptor P2Y12 and GPR34, a receptor for lysophosphatidylserine. While GPR34 deficiency has been linked to impaired microglial phagocytosis, its regulation in relation to amyloid-β (Aβ) and tau pathology in Alzheimer's disease (AD) remains unclear. We performed a quantitative analysis of microglial density, morphology, and GPR34 expression in the medial temporal lobe cortex (MTLC) of elderly human body and tissue donors across the AD spectrum. Using fluorescence in situ hybridization and immunolabeling, we analyzed 187,670 microglial cells and correlated microglial parameters with the severity and spatial proximity of Aβ plaques and tau inclusions. In parallel, we analyzed human single-nucleus RNA sequencing data from 236,002 cells to assess GPR34 expression across microglial subtypes, brain regions, and neuropathological stages. Microglial density and overall morphology in the MTLC were largely preserved, independent of local Aβ or hyperphosphorylated tau burdens. Apart from a moderate shortening of microglial processes in the immediate vicinity of Aβ plaques, no consistent pathology-associated morphological changes were detected. GPR34 expression showed pronounced cell-to-cell variability and differed across microglial subtypes and brain regions, but neither expression intensity nor the proportion of GPR34-positive microglia correlated consistently with Braak stage or Thal phase. These findings suggest that GPR34 regulation in human microglia is highly context-dependent and shaped by regional and cellular heterogeneity rather than AD-associated pathology alone.
The heart of Phasmida (stick insects) remains poorly understood, particularly regarding its overall physiology and associated cells. Here, we integrate the morphology of the heart and associated cells in Cladomorphus phyllinus with transcriptomic profiling of corresponding structures in Cladomorphus trimariensis. The heart is a tubular structure composed of cardiomyocytes physically associated with alary muscles, with a location and structure similar to that described in other insects. Numerous pericardial cells are organized in cord-like aggregations around the heart, displaying abundant membrane invaginations and an extensive endocytic and lysosomal machinery, consistent with their role in hemolymph filtration. Given their abundance, these cells likely represent the main source of the numerous transcripts related to endocytosis, vesicular trafficking and lysosome-mediated digestion. Hemocytes are also present around the heart, often forming aggregations indicative of immune responses. The transcriptomic analysis indicates immune activity, including melanization and antibacterial defense, predominantly driven by the hemocytes, with the pericardial cells providing a supportive role. Our results confirm that heart and associated cells act not only in hemolymph propulsion, but also as a site involved in maintaining hemolymph homeostasis and immune defense. This work provides an integrated morphological and functional framework for the circulatory system of Phasmida, expanding comparative perspectives across insects.
Colorectal cancer is one of the most common malignancies worldwide, and microbiome research has strong potential to advance understanding of tumor biology and to support biomarker development and microbiome-targeted interventions. Most colorectal cancer microbiome studies rely on stool or mucosal sampling, but routine pathology archives contain abundant formalin-fixed, paraffin-embedded primary tumor tissue that could enable scalable assessment of tumor-associated microorganisms. We profiled tumor-associated microorganisms in primary colorectal adenocarcinoma specimens from 192 patients using a targeted quantitative PCR panel and evaluated associations with clinicopathological variables (tumor differentiation grade, primary tumor T category, and disease stage) using non-parametric tests, ordinal regression, regularized logistic regression, and within-panel profile ordination with permutation-based inference. The most consistent signals were linked to tumor differentiation rather than stage. Tumors in the G2-G3 versus G1 comparison showed higher total bacterial load and a higher detection frequency of Ruminococcus spp., with both associations remaining significant after false discovery rate control; regression analyses corroborated these findings. A regularized logistic model achieved moderate discrimination for high-grade (G2-G3) disease (mean area under the receiver operating characteristic curve ≈0.71) and yielded a compact signature dominated by total bacterial load and detection of Ruminococcus spp., with additional contributions from low-frequency taxa that warrant cautious interpretation. In contrast, models targeting primary tumor T category, invasiveness (T1-T2 vs T3-T4), or overall stage showed low discriminative performance (area under the curve ≤0.59), and within-panel profile distances did not reveal robust global separation of groups. Targeted quantitative PCR profiling of archived primary tumor tissue identifies reproducible microbiome signals that track tumor differentiation grade more strongly than stage, suggesting that tissue bacterial burden and selected taxa may reflect microenvironmental features associated with the G2-G3 versus G1 contrast. Broader tumor microbiome profiling should be required to capture diversity and refine clinically informative signatures.
Head and neck squamous cell carcinoma (HNSCC) remains an immunosuppressive and metabolically dysregulated malignancy, contributing to tumor progression and resistance to conventional therapies. Natural compounds offer a unique multi-target opportunity to address these challenges, with berry-derived phytochemicals emerging as particularly promising candidates. Preclinical evidence demonstrates that these compounds modulate dendritic cell activation, macrophage polarization, regulatory T cell function, and cytokine signaling, restoring immune balance while simultaneously regulating tumor metabolism and reducing chronic inflammation. Beyond these immunometabolic effects, berry-derived compounds influence glucocorticoid signaling at the endocrine-immune interface, alleviating additional immunosuppressive pressures within the tumor microenvironment. Early clinical studies support the feasibility of standardized berry-derived formulations as adjunctive agents. In patients with oral premalignant lesions and HNSCC, black-raspberry-based interventions including topical gels and oral troches, have demonstrated favorable safety profiles, measurable tissue uptake of bioactive phytochemicals, modulation of proliferation and inflammation-associated biomarkers (e.g., Ki-67, COX-2, and NF-κB), and partial histologic regression in a subset of lesions. Collectively, these pleiotropic actions highlight chemopreventive potential and provide a mechanistic rationale for combinatorial strategies with immune checkpoint inhibitors targeting PD-1/PD-L1 and CTLA-4. Opportunities for both local and systemic delivery may further enhance therapeutic efficacy. Integrating these natural compounds into precision chemoprevention and immunotherapy paradigms could inform rational drug discovery, biomarker-driven patient stratification, and combination therapy design. This review highlights the convergent immunologic, metabolic, and endocrine-targeted mechanisms of berry-derived phytochemicals in HNSCC and emphasizes their translational potential as integrative modulators of antitumor immunity.
Salmonella enterica is a gastrointestinal pathogen that causes a variety of intestinal and systemic clinical disease. Vaccine development for systemic salmonellosis requires a deeper understanding of protective mechanisms and the relative contribution of mucosal and systemic host responses to bacterial elimination. Here, we examined the relative importance of mucosal and systemic immunity in the control of systemic salmonellosis. Delivery of a live-attenuated vaccine strain via a parenteral route did not elicit a substantial mucosal immune response, but was still sufficient to protect mice from systemic Salmonella infection. A prominent feature of the protective systemic response was the establishment of Salmonella-specific tissue resident memory (TRM) CD4 T cells in the liver. Although adoptive transfer of hepatic TRMs was sufficient to protect mice from challenge infection, TRMs from the intestinal lamina propria reduced bacterial loads but failed to fully protect mice from infection. The protective contribution of liver and LP TRMs was partially dependent on the expression of IL-18R and the corresponding ability to elicit noncognate CD4 T cell responses in vivo. Overall, these data suggest that a parenteral vaccination approach that efficiently induces liver TRMs expressing IL-18R should provide robust protection against mucosal or systemic infection with Salmonella.