The Seventh Affiliated Hospital of Sun Yat-sen University

Although endogenous neural stem cells represent a promising target for noninvasive spinal cord injury repair, inflammatory lesion environments frequently trigger their death. Our prior work identified necroptosis as a key death pathway for endogenous neural stem cells migrating to spinal cord injury lesions. Rapamycin-insensitive companion of mTOR (Rictor; a core component of the mechanistic target of rapamycin complex 2 [mTORC2] complex) regulates neural stem cell self-renewal and differentiation, and our preliminary data implicate it in spinal cord injury repair; however, its role in promoting endogenous neural stem cell survival post-spinal cord injury remains unclear. Here, we generated conditional endogenous neural stem cell-specific Rictor knockout mice using the Cre-loxP system. Although the endogenous neural stem cell-specific Rictor knockout mice displayed normal baseline spinal cord morphology and function, they exhibited impaired functional recovery after spinal cord injury compared with wild-type controls. This deficit correlated with elevated inflammatory responses and the increased susceptibility of endogenous neural stem cells to necroptosis. Mechanistically, lentiviral-mediated Rictor knockdown in neural stem cells in vitro impaired lysosomal function, leading to heightened sensitivity to tumor necrosis factor-alpha- and lipopolysaccharide-induced necroptosis. Collectively, these findings indicate that Rictor/mTORC2 signaling protects endogenous neural stem cells against receptor-interacting protein kinase 1-mediated necroptosis following spinal cord injury. Consequently, the modulation of intrinsic Rictor activity represents a potential therapeutic strategy to enhance endogenous neural stem cell survival and functional recovery post-spinal cord injury.

Conventional chemotherapy is significantly hampered by the inherent hydrophobicity of chemotherapeutic agents, limited tumor-specific targeting, and inadequate intratumoral accumulation, all of which undermine its clinical efficacy. Nonselective distribution of cytotoxic agents leads to suboptimal drug concentrations within tumor tissues, causing systemic toxicity in healthy organs. Tumor microenvironment-responsive nanoplatforms offer a promising strategy for enhancing specificity and efficacy. This study demonstrates the successful development of a nanodrug delivery system, CASS@PTX nanoparticles, where CASS is a cinnamaldehyde-based, disulfide-containing polymer engineered with dual-stimulus responsiveness to glutathione (GSH) depletion and reactive oxygen species (ROS) amplification. This system disrupts intracellular redox homeostasis in tumor cells, triggering the release of encapsulated paclitaxel (PTX) while enhancing chemotherapeutic efficacy through redox-dependent sensitization. GSH consumption and ROS overproduction create a prooxidative microenvironment that enhances PTX-induced apoptosis. Preclinical validation using in vitro cytotoxicity assays and in vivo tumor models demonstrates potent synergistic anti-tumor effects with minimal systemic toxicity. This cascading ROS self-generation strategy represents a promising approach for overcoming multidrug resistance and improving the therapeutic outcomes of cancer chemotherapy.

While lipoprotein apheresis (LA) is established for lipid management in primary biliary cholangitis, the role of apheresis techniques in drug-induced cholestatic liver disease remains unexplored. We report the novel application of double filtration plasmapheresis (DFPP) for rapid reversal of severe hypercholesterolemia in iatrogenic cholestasis, where conventional therapies failed or were contraindicated. A 56-year-old male developed severe cholestatic hepatitis following the use of herbal agents. Laboratory investigation revealed extreme hypercholesterolemia, with a total cholesterol (TC) of 41.39 mmol/L and a low-density lipoprotein cholesterol (LDL-c) of 38 mmol/L. Standard medical therapy proved ineffective. Two sessions of DFPP resulted in > 90% reduction in cholesterol levels and simultaneous improvement in bilirubin and liver enzymes, with sustained normal levels at 1-year follow-up. DFPP may serve as a novel rescue therapy for drug-induced cholestatic hypercholesterolemia, providing simultaneous removal of cholesterol and bilirubin where other treatments are ineffective or at high risk.

Neonatal hypoxic-ischemic encephalopathy (HIE) may induce substantial neuronal damage. In particular, an overactivation of microglia following HIE represents a pathogenically important process. Previous studies have shown that microglial inhibitors can exert neuroprotective effects in HIE; however, the specific mechanisms underlying these effects have not yet been elucidated. Ligation of the left common carotid artery and exposure to 5% O2 were utilized to produce an HIE model in rats. A number of experimental approaches were then used to determine the effect of a microglial inhibition, achieved via the administration of GW2580, a Csf1r inhibitor, and investigate the mechanisms involved. Our HIE models exhibited substantial brain infarction and were significantly impaired in motor functions (p < 0.01-0.001, in all tests examined). In the infarction areas, the number of microglia, macrophages, and neural stem cells (NSCs) was all dramatically increased over that in sham-injured rats, respectively (p < 0.05-0.001). The administration of GW2580 significantly reduced the numbers of microglia and macrophages, but increased the number of NSCs when compared to those in vehicle-treated HIE models (p < 0.05-0.001). Furthermore, GW2580 significantly ameliorated both the histological and behavioral phenotypes in HIE rats and increased STAT phosphorylation (p < 0.05-0.001). Finally, the inhibition or activation of STAT3 respectively decreased or increased the neuroprotective effects of GW2580 (p < 0.05-0.001). Collectively, our findings demonstrate that the STAT3 signaling pathway plays a critical role in the neuroprotective effects of microglial inhibition and may facilitate the development of novel therapeutic strategies to treat stroke.

Abstract Background Pancreatic ductal adenocarcinoma (PDAC) remains one of the most aggressive human malignancies, characterized by a five-year survival rate of approximately 13 percent and limited therapeutic efficacy. Although Claudin18.2 (CLDN18.2)-directed chimeric antigen receptor (CAR) T cells have demonstrated promising antitumor activity, physiological CLDN18.2 expression in normal gastric epithelium poses a risk of severe on-target, off-tumor toxicity. Methods An AND-gated CAR platform incorporating LAT and SLP-76 signaling modules was generated to restrict T-cell activation to concurrent recognition of CLDN18.2 and Mesothelin (MSLN). Antitumor efficacy, specificity, and safety were evaluated using PDAC cell lines and murine PDAC xenograft models. Durability-associated transcriptional programs and functional stability were assessed by transcriptomic profiling with quantitative PCR validation and multi-round killing assays. Results The AND-gated CAR-T cells exhibited stringent antigen dependency, selectively eliminating dual-antigen-positive tumor cells while sparing single-antigen or low antigen density targets. In addition, transcriptomic profiling revealed enrichment of memory- and durability-associated programs in AND-gated CAR-T cells, which was supported by increased expression of C-X-C motif chemokine receptor 6 (CXCR6), interleukin-7 receptor (IL7R), and C-C motif chemokine ligand 5 (CCL5), along with reduced expression of interleukin-2 receptor alpha (IL2RA) and nuclear receptor subfamily 4 group A member 1 (NR4A1) in both CD4⁺ and CD8⁺ subsets. Consistent with this profile, AND-gated CAR-T cells maintained cytotoxic activity across repeated antigen challenges, whereas conventional CLDN18.2 CAR-T cells exhibited progressive functional decline. Importantly, in murine PDAC models, AND-gated CAR-T cells achieved pronounced tumor regression while preserving gastric tissue integrity and without overt toxicity, corroborating their translational potential. Conclusions These findings demonstrate that AND-gated CLDN18.2&amp;MSLN CAR-T cells achieve tumor-restricted cytotoxicity in PDAC while exhibiting durable antitumor function and transcriptional programs associated with T-cell functional stability, supporting AND-gated CAR-T strategies to improve the safety and durability of CAR-T therapy in solid tumors.

This study aims to develop and validate a nomogram model based on surgical parameters and clinical features for predicting the risk of poor pulmonary rehabilitation outcomes in patients with non-small cell lung cancer (NSCLC) after radical surgery. This retrospective cohort study included 320 patients who underwent radical lung cancer surgery between June 2021 and December 2024. Patients were divided into a development cohort (n = 224) and a validation cohort (n = 96) at a 7:3 ratio. A composite surgical trauma score (STS) was developed to integrate key intraoperative variables. Independent predictors were identified through multivariate logistic regression analysis, including surgical trauma score, operative time, intraoperative blood loss, and other clinical factors. A nomogram model was constructed and validated using the bootstrap resampling (1000 repetitions). Model performance was assessed using the area under the curve (AUC) and calibration metrics. The incidence of poor pulmonary rehabilitation was 38.4% in the development cohort and 36.5% in the validation cohort. In the development cohort, 11.2% of patients had a forced expiratory volume in 1 second (FEV1) recovery rate <80% alone, 15.2% had a modified Medical Research Council (mMRC) score ≥2 alone, and 12.0% met both criteria. Multivariable analysis predicted five independent variables: high versus low surgical trauma score (odds ratio (OR) = 3.09, 95% confidence interval (CI): 1.71-5.58), preoperative FEV1% predicted <70% (OR = 2.67, 95% CI: 1.53-4.67), operative time ≥180 minutes (OR = 2.52, 95% CI: 1.41-4.50), intraoperative blood loss ≥80 mL (OR = 2.25, 95% CI: 1.27-3.99), and age ≥65 years (OR = 1.92, 95% CI: 1.10-3.35). The nomogram demonstrated good discrimination, with an AUC of 0.84 (95% CI: 0.78-0.90) in the development cohort and 0.79 (95% CI: 0.70-0.88) in the validation cohort. Calibration was satisfactory in both cohorts (p = 0.23 in the development cohort; p = 0.31 in the validation cohort). Decision curve analysis revealed meaningful net benefits across a wide range of threshold probabilities. Predictive performance remained consistent across subgroups defined by age groups, surgical trauma strata, and tumor-node-metastasis (TNM) stages. Furthermore, sensitivity analyses using alternative endpoint definitions and various approaches for handling missing data further confirmed the robustness of the model. We developed and validated a predictive nomogram for poor pulmonary rehabilitation outcomes after radical lung cancer surgery. This model incorporates readily available clinical and surgical parameters, providing individualized risk assessment that may facilitate personalized rehabilitation strategies and improve postoperative management.