The transformation of lung adenocarcinoma into large cell neuroendocrine carcinoma (LCNEC) in terms of genotype and histology has been described as a mechanism of acquired resistance to epidermal growth factor receptor tyrosine kinase inhibitor (EGFR-TKI). However, this phenomenon is exceedingly rare in anaplastic lymphoma kinase (ALK)-positive lung adenocarcinoma. Here, we report a case of an ALK-positive lung adenocarcinoma patient who developed resistance following sequential treatment with the ALK-TKI alectinib and lorlatinib, accompanied by histological transformation to LCNEC and concurrent genetic alterations including TP53 deletion, CDKN2A deletion, and MYC amplification. This case expands the spectrum of ALK-TKI resistance mechanisms and highlights the potential value of exploring combinatorial approaches incorporating immunotherapy, antiangiogenic therapy, and chemotherapy for the management of such cases.

Small cell lung cancer (SCLC) accounts for approximately 15% of all lung cancer cases. SCLC is characterized by rapid proliferation, early distant metastasis, high recurrence rate, and poor prognosis, with a 5-year survival rate of less than 7%. Immune checkpoint inhibitor (ICI)-based immunotherapy has been widely applied in SCLC clinical practice, including immunotherapy combined with chemotherapy as first-line therapy, tarlatamab (post-chemo/immunotherapy) and lurbinectedin (post-platinum chemotherapy) as later-line options for extensive-stage SCLC (ES-SCLC), and chemoradiotherapy (CRT) followed by consolidation of immune therapy for limited-stage SCLC (LS-SCLC). However, the efficacy of immune combination therapy in SCLC patients remains limited and studies confirmed that objective response rate (ORR) of first-line immunochemotherapy in advanced SCLC is approximately 60-70%, while the durable response rate remains only 10-20%. The main reason lies in the complex and heterogeneous immune microenvironment of SCLC, where a network of immunosuppressive factors orchestrates an immune-excluded or "cold" phenotype. With the development of genomics application in SCLC research, increasing data have revealed and validated the immune microenvironment features of SCLC. However, there is a lack of systematic reviews in this field. This review focuses on clinical translational research across SCLC stages and subtypes, systematically summarizing the immune microenvironment features of LS-SCLC, ES-SCLC, recurrent/relapse SCLC, and never-smoker SCLC (nsSCLC) by integrating data generated from genomic sequencing, single-cell sequencing, spatial transcriptomics, and other research approaches. We summarize the biological and immune features of SCLC and highlight key biomarkers, with the goal of offering new perspectives to improve immunotherapy outcomes and uncover novel therapeutic vulnerabilities.

Background:RET fusion is a pathogenic driver factor in lung cancer patients. Currently, the conclusions on the clinical factors of RET fusion in NSCLC are inconsistent. Methods:From 2018 to 2024, 6,204 lung cancer patients received next‑generation sequencing (NGS) testing, among whom 102 were confirmed to be positive for RET fusion. The clinical and molecular characteristics of these patients were analyzed and compared. Results:In this cohort, the prevalence of RET fusions was 1.6% (102/6204). Most patients were female (54.90%), <60 years (53.92%), non-smokers (72.55%), with advanced-stage (68.63%), metastatic (69.61%, mostly lymph nodes), adenocarcinoma (98.04%), and PS 0-1 (90.20%). The most common fusion partners of RET were KIF5B (50.00%, 51/102) and CCDC6 (22.55%, 23/102).; noval partners including C16orf95, CARNMT1-AS1, CXCL12, MTUS1 and MYRFL were identified. Common fusion partners were associated with age (P=0.023) and PS score (P=0.040), with higher rates of RET fusion in patients <60 years of age and those with a PS score of 0-1 (81.80% and 76.10%, respectively. TP53 represented the most frequent concomitant alteration in RET fusions, occurring at a rate of 14.71% (15/102). Conclusion:The new discoveries of RET fusion partners were founded in NSCLC. In addition, the broad-panel NGS is essential for NSCLC patients to catch these rare/novel fusions that PCR or small panels might miss.

PURPOSE:To delineate age-related changes in visual, structural, and electrophysiologic measures in X-linked retinoschisis (XLRS) and to assess automated cyst-cavity volume (CCV) as a quantitative biomarker. DESIGN:Single-center mixed retrospective-prospective cohort study. METHOD:This study included 440 visits from 107 individuals with clinically diagnosed XLRS and confirmed pathogenic RS1 variants. Comprehensive ophthalmic evaluations were performed at baseline and during follow-up, including visual acuity (VA), fundus examination, electroretinogram (ERG), and spectral-domain optical coherence tomography (SD-OCT), with microperimetry (MP) conducted in a subset of patients. Among SD-OCT parameters, CCV was automatically quantified using self-developed DEEP-OCT-CCSEG system. Annual progression rates were estimated using restricted cubic spline or linear regression models, and interocular agreement was evaluated with Bland-Altman and intraclass correlation analyses. RESULTS:In this cohort, 70% patients presented by age 10. The median age at first examination was 7.3 years (range, 1.9-78.5) over a mean follow-up of 1.9 years. Baseline means were: VA 0.82 ± 0.47 logMAR, central foveal thickness (CFT) 495 ± 186 µm, and CCV 1.54 ± 1.16 mm³. Genetic analysis identified 64 distinct RS1 variants, including 5 that were novel. Longitudinally, visual acuity followed a triphasic pattern: slight improvement in childhood, a prolonged plateau through mid-adulthood, and a more rapid decline thereafter, with Kaplan-Meier analysis projecting 40% of patients will be blind by age 60. Scotopic ERG a- and b-wave amplitudes peaked in early childhood before declining with age, while the b/a ratio gradually decreased. In OCT, CFT showed a modest, significant age-related decrease. In contrast, CCV followed a non-linear age-related pattern, increasing to a peak in the third decade of life before progressively declining. CONCLUSIONS:In this XLRS cohort, most patients showed gradual structural and functional decline beginning in the second decade of life, suggesting an optimal therapeutic window within early adulthood. By applying CCV as a quantitative biomarker in a large-scale longitudinal setting for the first time, this study quantitatively characterized 3-dimensional retinal remodeling and delineated age-related structural changes in XLRS. These findings refine the understanding of XLRS natural history and provide a robust framework for therapeutic evaluation and clinical trial design.

Hypoxia exerts a far-reaching influence on cellular metabolic activities, which in turn has profound implications on cell functions. Emerging evidence has demonstrated that cell-free mitochondrial DNA (cf-mtDNA) exhibits a prompt response to acute stress within a relatively short span. Nevertheless, to date, there is a lack of reports on how hypoxic stress affects cf-mtDNA. Hypoxic stress cell models and mouse models were established. Western blot was used to examine the expression levels of HIF-1α and HIF-2α protein, and droplet digital PCR (ddPCR) was used to determine the copy numbers of cf-mtDNA in cell supernatants or in mice sera. Exosomes were isolated from the cell supernatants, with their characteristics analyzed by electron microscopy, nanoparticle tracking analysis (NTA), and Western blot. ELISA method was used to analyze the cortisol levels in mice sera. Receiver Operating Characteristic (ROC) curve was established to interpretate the ability of cf-mtDNA evaluating hypoxic stress. The cf-mtDNA copy number alterations are associated with hypoxic stress. Cf-mtDNA levels were found to increase under hypoxic stress, which was primarily derived from exosomes. Cf-mtDNA and cortisol levels exhibited an ascending tendency in hypoxic mice sera, with the AUC values for cf-mtDNA being 0.8690 on Day 3 and 0.8976 on Day 7, indicating that cf-mtDNA could serve as a potential predictor for hypoxic stress. The copy numbers of cf-mtDNA were increased under hypoxic stress. The extracellular release of cf-mtDNA was mainly derived from exosomes. Cf-mtDNA was proposed as a potential diagnostic predictor for acute mild hypoxic stress.

Purpose:To characterize retinal structural alterations and elucidate underlying pathogenesis in female carriers of choroideremia (CHM) using multimodal imaging. Methods:This single-center retrospective study analyzed 100 eyes of 50 genetically confirmed female CHM carriers. Clinical evaluation comprised genotype, age, visual acuity, color fundus photography, fundus autofluorescence (FAF), infrared reflectance, and optical coherence tomography (OCT). Retinal lesions were identified and classified using point-to-point co-localization analysis across modalities. Results:The mean age of carriers was 39.6 ± 17.8 years (range 3.2-80.3 years). All eyes exhibited varying degrees of retinal abnormalities. Most common findings included yellowish changes on color photographs, mottled areas of hyper- and hypo-autofluorescence on FAF, and isolated interdigitation zone (IZ) loss on OCT. Chorioretinal atrophy beyond the peripapillary region, indicating a severe phenotype, was observed in 18% of eyes. Five characteristic degenerative lesions were identified: Type A, isolated IZ loss; Type B, hyper-autofluorescent spots corresponding to presumed photoreceptor loss; Type C, drusen-like deposits; Type D, degeneration of the retinal pigment epithelium (RPE) and RPE-photoreceptor interface; and Type E, outer retinal degeneration. Type B lesions suggest that photoreceptor loss may precede RPE loss at specific loci. Type A lesions were the earliest abnormality, significantly associated with milder phenotypes, indicating the RPE-photoreceptor interface is the initial site of pathology. In contrast, Type D and E lesions were associated with severe phenotypes. Conclusions:Identified hyper-autofluorescence lesions corresponding to presumed photoreceptor loss indicate independent degeneration of photoreceptors and RPE. These findings suggest that future therapeutic approaches should concurrently target both cellular components.

Astrocytes act as crucial cellular centres of cholesterol synthesis and metabolism and help maintain homeostasis in the healthy CNS. Spinal cord injury (SCI) results in abnormalities in astrocytic cholesterol metabolism and excessive oxysterol accumulation, contributing to the activation of inflammation. However, the relevant regulatory mechanism involved in aberrant cholesterol metabolism by astrocytes has not been fully elucidated. In the present study, we demonstrated that SCI-induced d-DT protein levels increased synchronously with CH25H expression. Administration of the d-DT inhibitor 4-CPPC markedly decreased CH25H expression in astrocytes following SCI. d-DT facilitates CH25H production in astrocytes by activating the intracellular ERK/NF-κB pathway through binding to the CD74 receptor. Conditioned culture medium from astrocytes following the knockdown of astrocyte CH25H expression by siRNA reduced microglial migration. The inhibition of d-DT or CH25H activity reduces microglia/macrophage accumulation at the lesion site and improves motor functional recovery following SCI. Our results reveal a novel function of d-DT-mediated astrocytic CH25H activation, which modulates pathological microenvironments through the activation of inflammation. These data may provide a potential therapeutic strategy for CNS inflammation-associated diseases.

Choroideremia is an X-linked chorioretinal dystrophy with well-characterized progression in affected males but variable phenotypes in female carriers. Understanding the phenotypic spectrum in female carriers is important for prognosis, monitoring, and trial design. This study aims to delineate the natural history of retinal phenotypes and visual function loss in female choroideremia carriers and establish an improved fundus grading system for disease stratification and prognostic prediction. This single-center, longitudinal and cross-sectional, retrospective study included 64 genetically confirmed female choroideremia carriers. Clinical data included genotype, age, best-corrected visual acuity, color fundus photography, fundus autofluorescence, visual field testing, and full-field electroretinography. A novel fundus phenotypic grading system was proposed based on fundus autofluorescence and fundus color photographs, which included four types: granular (merged fine/coarse patterns), severe peripapillary atrophy (highlighting severe peripapillary atrophy as a crucial feature), localized atrophy, and widespread atrophy. The agreement between measurement-based grading and visual grading was assessed. Visual acuity and fundus phenotypes showed moderate interocular symmetry, while visual field and electroretinography metrics showed high interocular symmetry. At baseline, phenotypes included granular (76.3

BACKGROUND:Small cell lung carcinoma (SCLC) is a highly aggressive, rapidly proliferating malignancy largely corresponding to immune-cold tumor type. The role of the key oncogene c-Myc in regulating the complex immune microenvironment of SCLC remains unknown. METHODS:Immunohistochemistry and multiplex immunofluorescence techniques were used to analyze c-Myc expression and immune cell infiltration. c-Myc and β-catenin expression were analyzed using western blotting. Flow cytometry was used to quantify the expression of immunosuppressive ligands-including programmed death ligand 1 (PD-L1), CD47, CD155, and human leukocyte antigen-E (HLA-E)-and characterize the function of T cells and macrophages. Cytokine levels were evaluated using enzyme-linked immunosorbent assays. RESULTS:c-Myc expression in high tumor-infiltrating lymphocyte infiltration (TILhigh) tumor exceeded that in TILlow tumors (n = 24) and was positively correlated with CD163+ macrophage infiltration (p = 0.0180). Immunosuppressive ligands were universally expressed on SCLC cells. 10058-F4-a c-Myc inhibitor-substantially downregulated the expression of PD-L1, CD47, and CD155 and upregulated HLA-E expression. In the coculture system, the inhibition of c-Myc in SCLC cells significantly upregulated interferon-gamma production in Jurkat cells, inhibited macrophage phagocytosis, and decreased cytokine concentrations in the supernatant. Further mechanistic studies revealed that c-Myc inhibition modulated the Notch signaling pathway and reduced the expression of E-cadherin, vascular endothelial (VE)-cadherin, and β-catenin. A β-catenin agonist improved the regulatory effect of 10058-F4 on the immunosuppressive ligands in SCLC cells. CONCLUSIONS:c-Myc expression in SCLC is associated with immune infiltration and may regulate the expression of immune-related ligands on SCLC cells via WNT/β-catenin signaling.

Esophageal squamous cell carcinoma (ESCC) is a highly prevalent and aggressive malignancy worldwide, associated with poor prognosis. Most patients are diagnosed at an advanced stage, where conventional chemotherapy offers limited therapeutic efficacy and is often accompanied by substantial toxicity. In recent years, immune checkpoint inhibitors (ICIs), particularly those targeting PD-(L)1 and CTLA-4, have emerged as cornerstone therapies in both first-line and subsequent treatment settings for advanced ESCC. Nevertheless, significant clinical challenges persist, including the complexity of mechanisms underlying immune resistance, suboptimal predictive performance of existing biomarkers, difficulties in the management of immune-related adverse events (irAEs), and underrepresentation of elderly patients in clinical trials. This review summarizes recent advances in immunotherapy for advanced ESCC, evaluating the clinical evidence supporting ICIs as monotherapy or in combination with agents such as anti-angiogenic drugs and tyrosine kinase inhibitors. It further discusses the therapeutic potential of novel approaches, including bispecific antibodies, CAR-T cell therapy, and next-generation ICIs, while addressing current treatment paradigms for elderly patients. The importance of comprehensive, longitudinal management of irAEs is emphasized. Additionally, this article provides an in-depth analysis of mechanisms contributing to immune resistance—such as loss of tumor neoantigens and dysregulation of key signaling pathways—and critically appraises the limitations of established biomarkers, including PD-L1 expression and tumor mutational burden (TMB), alongside emerging developments in biomarker discovery. In conclusion, while immunotherapy has significantly improved outcomes and expanded therapeutic prospects for patients with advanced ESCC, further research is required to elucidate resistance mechanisms, refine treatment strategies, and identify robust predictive biomarkers. These efforts are essential to advance precision medicine in ESCC and ultimately enhance long-term survival outcomes.

Objective To investigate the clinical characteristics and impact of SMARCA4 mutations in patients with non-small cell lung cancer (NSCLC).Methods A total of 2,821 patients with NSCLC who underwent next-generation sequencing were retrospectively included. The frequency and types of SMARCA4 mutations and co-mutations were determined, and the clinical outcomes were assessed.Results SMARCA4 mutations were identified in 100 samples (3.54%), and 36% were missense mutations. The most frequent co-mutations were TP53 (67%) and EGFR (31%); 13% of SMARCA4 mutations occurred in samples carried EGFR and TP53 mutations. Notably, 63% SMARCA4 mutations did not present druggable driver mutations. SMARCA4 mutations were most prevalent in males and smokers. Patients with SMARCA4 mutant lung adenocarcinoma (LUAD) and EGFR mutations who received EGFR-tyrosine kinase inhibitors (EGFR-TKI) as first-line therapy had a lower objective response rate (ORR, 52.94%). In SMARCA4 mutation and EGFR wild-type (wt) NSCLC cohort who received first-line chemotherapy, age (hazard ratio [HR], 3.090; p = 0.026) and performance score (HR, 5.848; p = 0.045) were identified as independent predictors of progression-free survival (PFS). Conversely, brain metastasis was an independent predictor of superior overall survival (HR, 0.188; p = 0.011). The patients with EGFR wt and SMARCA4 mutant Stage IV LUAD who received chemotherapy plus anti-angiogenic therapy significantly improved median PFS compared to chemotherapy alone (p = 0.04).Conclusions SMARCA4 mutations were predominantly males and smokers in NSCLC. SMARCA4 mutations conferred a poorer response for EGFR-mutant LUAD subgroups who received EGFR-TKIs. Additionally, chemotherapy plus anti-angiogenesis as first-line therapy may be more effective for Stage IV-SMARCA4 mutant LUAD with EGFR wt.

Elderly patients exhibit heightened susceptibility to postoperative complications following general anesthesia and surgery, yet the molecular mechanisms driving this age-dependent vulnerability remain poorly defined. We performed RNA sequencing on olfactory bulb (OB), hippocampus (HI), lung, and spleen from young (3-month, m), late middle-aged (17 m), and geriatric (27 m) male C57BL/6 mice 24 h after 2 h of exposure to isoflurane anesthesia and laparotomy (ISO/OP). Short-term ISO/OP elicited pronounced, age-dependent transcriptional remodeling across tissues. Late middle-aged mice exhibited robust activation of stress- and metabolism-associated pathways in the OB and HI, accompanied by suppression of lipid, synaptic, and structural maintenance programs. In contrast, young adults displayed limited responses, characterized by modest and adaptive synaptic remodeling in the HI. Peripheral organs showed a parallel age-dependent divergence. Late middle-aged mice exhibited amplified immune and inflammatory signaling in the lung and spleen alongside suppression of structural, regulatory, and metabolic homeostatic programs, whereas young adults demonstrated attenuated, metabolically adaptive transcriptional responses. Circulating extracellular vesicles (EVs) mirrored tissue-level shifts, indicating a systemic transition from adaptive plasticity in 3 m to stress and immune dominant signaling by 17 m. Geriatric mice displayed a distinct response pattern, characterized by activation of stress and detoxification programs in brain tissues, altered circadian gene expression in lung and spleen, and extensive remodeling of EV protein cargo enriched for inflammatory and growth factor-related signatures. Together, these findings indicate that late middle-age is associated with amplified peri-anesthetic biological reactivity across central and peripheral systems, suggesting an under-recognized window for perioperative risk stratification and preventative intervention.

Cryopreserved peripheral blood mononuclear cells (PBMCs) are widely used in RNA sequencing (RNA-seq), and the quality of their RNA directly influences test outcomes. However, cryopreservation can readily lead to RNA degradation in PBMCs, and the issue of how to improve RNA quality during long-term cryopreservation remains unresolved. PBMCS from fresh blood samples collected from patients with advanced non-small cell lung cancer (NSCLC) were utilized as controls. The RNA preservation efficacy of PBMC samples was compared between the classical cell cryopreservation solution (90% fetal bovine serum [FBS] + 10% dimethyl sulfoxide [DMSO]) and the common RNA extraction reagent (TRIzol) at -80 ˚C for 1, 3, and 6 months. RNA concentration, purity, and integrity (as measured by the RNA integrity number [RIN]) were assessed using a multi-sample microvolume UV-Vis spectrophotometer and an automated microfluidic electrophoretic bioanalyzer to provide a theoretical foundation for the preservation of high-quality RNA samples. This study demonstrated that the concentration, purity, and integrity of RNA in PBMCs decreased with extended cryopreservation duration. While the two processing methods had no effect on RNA concentration, they did affect RNA purity and integrity. The RNA integrity of PBMCs cryopreserved with TRIzol for 6 months remained satisfactory. Our results indicate that TRIzol contributes to effective cryopreservation and maintains high-quality RNA.

Small-cell lung cancer (SCLC) is widely recognized as a notoriously refractory tumor, characterized by rapid proliferation, high aggressiveness, and early metastasis. Although immunotherapy combined with chemotherapy has become the standard of care for patients with extensive-stage SCLC, significant unmet needs, especially referring to lack of targeted therapy, persist in the clinic. The c-MYC proto-oncogene, one of the critical oncogenes, has been shown to be heavily associated with malignant tumor growth and drug resistance. c-MYC is amplified in SCLC and is further implicated in SCLC phenotypic plasticity, immune microenvironment composition, metabolic reprogramming, and therapy vulnerability, thereby playing the most key role in SCLC recurrence and devastating survival outcomes. This review summarized current knowledge on the mechanisms by which c-MYC regulates the SCLC oncogenic network and the status of drug development efforts targeted c-MYC. We also explore in detail that targeting c-Myc protein directly or indirectly represents a promising strategy for novel SCLC therapies.

PURPOSE:To characterize the clinical and genetic spectrum of TTLL5-related retinal dystrophy. DESIGN:Retrospective observational study. SUBJECTS:Twenty-one affected individuals from 19 unrelated families carrying biallelic TTLL5 variants. METHODS:Patients with inherited retinal dystrophy and confirmed biallelic TTLL5 variants were retrospectively reviewed. Genetic diagnosis was established using whole-exome sequencing followed by Sanger confirmation and cosegregation analysis. Clinical assessments included best-corrected visual acuity, multimodal retinal imaging, visual field testing, and full-field electroretinography. MAIN OUTCOME MEASURES:Visual function, retinal structural and functional characteristics, and genotype-phenotype correlation. RESULTS:The cohort included 10 males and 11 females, aged 6 to 66 years. Based on phenotypic classification, 2 patients were diagnosed with cone dystrophy (CD), 12 with cone-rod dystrophy (CRD), and 7 with rod-cone dystrophy (RCD). Best-corrected visual acuity ranged from 0 to 2.70 logarithm of the minimum angle of resolution (LogMAR) (median, 0.6 LogMAR; interquartile range, 1.55) and was significantly correlated with age (Spearman R = 0.704; P < 0.001). All patients were myopic, with spherical equivalent values ranging from -1.13 to -18.00 diopters, and 14 of 21 (66.67%) had bilateral high myopia (≤-6.0 diopters). Retinal involvement was characterized by macular degeneration with progressive extension beyond the posterior pole. Fundus autofluorescence demonstrated parafoveal or macular hyperautofluorescent rings followed by expanding hypoautofluorescent areas, whereas OCT revealed parafoveal-to-diffuse disruption or loss of the ellipsoid zone with outer nuclear layer thinning. Genetic analysis identified 24 pathogenic variants, of which 19 (79.17%) were novel. The recurrent missense variant c.437G>A (p.Gly146Glu) was the most frequent allele in this cohort and was consistently associated with a CRD phenotype. Extraocular manifestations were uncommon, with sensorineural hearing loss observed in 1 patient. CONCLUSIONS:TTLL5-associated retinal dystrophy exhibits a broad clinical spectrum encompassing CD, CRD, and RCD, with a high prevalence of myopia as a shared feature. The recurrent p.Gly146Glu variant showed a consistent association with the CRD phenotype, suggesting specific allele-phenotype correlations. These findings expand the recognized phenotypic and genetic spectrum of TTLL5-related disease and provide an important clinical framework for diagnosis, counseling, natural history, and future therapeutic studies. FINANCIAL DISCLOSURE(S):The authors have no proprietary or commercial interest in any materials discussed in this article.

Purpose:It is well established that volatile anesthetics and surgery induce acute and subacute changes in the cellular and molecular landscape of the brain and peripheral circulation and can impair neurological function. However, the chronic neurological sequelae of isoflurane (Iso) anesthesia combined with surgical operation (OP), as well as the underlying mechanisms of postoperative neurological dysfunction, remain poorly understood. Methods:Young adult male (M) and female (F) C57BL/6 mice underwent 4 h of 2% Iso plus laparotomy or sham treatment. At 12 weeks, olfactory and cognitive function were assessed by odor memory, buried food, Y-maze, and active avoidance tests. Olfactory bulbs (OB) and hippocampi (HI) were collected for RNAseq, while plasma extracellular vesicles (EVs) were isolated, characterized by NanoFCM, and profiled by Olink proteomics. Lastly, EVs were injected into the HI of naïve male mice, and cytokine/chemokine responses were measured 24 h later. Results:Both sexes showed olfactory impairment after chronic Iso/OP, with greater deficits in females. Iso/OP mice, especially females, exhibited impaired odor recognition in the OM test and longer latencies to locate buried food. Female mice also showed greater hippocampal-dependent spatial working memory deficits in the Y-maze, with more arm returns and fewer alternations than Sham/F mice, whereas Iso/OP/M mice performed similarly to controls. Likewise, female, but not male, Iso/OP mice displayed impaired associative learning in the active avoidance test, evidenced by fewer avoided trials and more escape responses. These long-term behavioral abnormalities were accompanied by sex-divergent transcriptomic remodeling in the OB and HI, including altered synaptic, neurodevelopmental, extracellular matrix, stress-response, and chemotaxis-related pathways. Iso/OP reduced plasma EV particle numbers in both sexes and shifted EV size distributions, with prominent reductions in the 40-100 nm EV fraction. Proteomics revealed distinct sex-and condition-specific EV profiles, with several EV-associated proteins showing opposing sex-dependent expression patterns. Hippocampal injection of Iso/OP-derived EVs induced donor sex-dependent cytokine remodeling, confirming inflammatory bioactivity. Conclusions:Four-hour isoflurane (Iso) exposure combined with laparotomy in young adult mice induces chronic, sex-dependent neurological deficits with distinct transcriptomic remodeling across brain subregions. Persistent alterations in circulating EV abundance and inflammatory cargo may drive chronic neuroinflammation and long-term brain dysfunction.

Background and Objective:Lung cancer remains the leading cause of cancer-related mortality worldwide. Despite advances in targeted and immunotherapy, challenges such as drug resistance persist. Traditional experimental models often fail to recapitulate human tumor complexity, limiting translational success. Patient-derived organoids (PDOs), which retain key characteristics of original tumors, have emerged as powerful tools. This review aims to systematically examine the methodologies, characterizations, and applications of lung cancer organoids (LCOs) in both basic and translational research. Methods:A narrative review was conducted based on literature retrieved from PubMed, Scopus, and Web of Science up to March 2026. Search terms included "lung cancer organoid/LCO", "patient-derived organoid/PDO", "three-dimensional (3D) culture", "2.5-dimensional (2.5D) culture", "drug susceptibility testing", and "precision medicine". Studies focusing on culture techniques, molecular characterization, and preclinical/clinical applications were included. Key Content and Findings:This study summarizes current LCO culture systems, comparing conventional 3D platforms and emerging 2.5D systems that offer significant advantages in cost, operational simplicity, and imaging convenience for rapid clinical applications. We detail multi-dimensional characterization approaches (morphological, molecular, functional) and discuss critical quality control standards. LCOs have been instrumental in studying tumorigenesis mechanisms, signaling pathways, and metabolic reprogramming. In translational research, LCOs show high predictive value for drug susceptibility to targeted agents, chemotherapy, and immunotherapy, and serve as platforms for developing novel therapeutics such as antibody-drug conjugates (ADCs). Conclusions:LCO models faithfully recapitulate tumor heterogeneity and are increasingly integral to precision medicine and drug development. While challenges remain in vascularization, immune microenvironment reconstitution, and standardization, ongoing integration with bioengineering and artificial intelligence (AI) promises to enhance their translational utility. This review underscores the potential of LCOs to bridge basic research and clinical practice, accelerating personalized therapeutic strategies in lung cancer.

Background: For limited-stage small cell lung cancer (LS-SCLC), immunotherapy combined with chemotherapy as induction followed by subsequent radiotherapy is still under clinical investigation. Some patients who receive treatment have undergone radiotherapy following immunochemotherapy in real-world. However, the preliminary efficacy and safety remain unknown. In this study, we will explore a novel treatment approach for LS-SCLC. Methods: This study analyzed 32 patients with LS-SCLC treated at Jilin Cancer Hospital between January 2022 and June 2024. Eligible patients received Immune checkpoint inhibitors (ICIs) combined with chemotherapy, followed by sequential or concurrent radiotherapy and maintenance chemo-immunotherapy. Tumor response was evaluated every two cycles per Response Evaluation Criteria in Solid Tumors (RECIST) v1.1, with progression-free survival (PFS) and overall survival (OS) as endpoints. Adverse events (AEs) were graded according to National Cancer Institute-Common Terminology Criteria (NCI-CTC) 5.0. Blood samples were collected at baseline and during treatment to assess immune function, neuron-specific enolase (NSE), pro-gastrin-releasing peptide (ProGRP), neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR), and lactic dehydrogenase (LDH). Results: All patients received a chemotherapy regimen of 4-6 cycles of carboplatin plus etoposide (EC) or cisplatin plus etoposide (EP). In the latest efficacy assessments, 3.1% achieved complete response (CR), 90.6% achieved partial response (PR), 6.3% achieved stable disease (SD), with overall response rate (ORR) of 93.7% and disease control rate (DCR) of 100%. Adverse reactions related to treatment included grade 3/4 myelosuppression occurring in 15.6%, grade 4 thrombocytopenia in 3.12% and radiation pneumonitis in 18.7%. Follow-up through June 2025 showed median PFS of 16.3 months [95% confidence interval (CI): 11.2-21.3] and median OS of 37 months (95% CI: 30.0-44.0) for first-line treatment. The incidence of abnormal baseline immune function was 78.9%, of which 80% presented as immune suppression. PLR demonstrated significant prognostic value: patients with baseline PLR >101.75 had longer PFS. Similarly, at the end of the second cycle or before radiotherapy, patients with PLR >60.1 also showed prolonged PFS. In contrast, dynamic changes in NLR did not demonstrate prognostic significance [hazard ratio (HR)=1.05, 95% CI: 0.32-3.53, P=0.93]. LDH testing suggested that patients with normal baseline LDH had longer PFS than those with elevated levels (18.5 vs. 10.0 months), although the difference was not statistically significant (P=0.15). Notably, in some patients, LDH levels decreased from elevated to normal during treatment, suggesting a potential association with treatment response. Conclusions: Chemotherapy combined with ICIs followed by sequential or concurrent radiotherapy is feasible, safe, and effective for LS-SCLC, achieving high short-term response rates and prolonged PFS. PLR strongly predicts PFS, while normal or decreasing LDH indicates favorable treatment response. Elevated NSE and ProGRP show trends toward shorter PFS. These biomarkers can guide prognosis and therapy monitoring, supporting personalized management in LS-SCLC.

Background:Colorectal cancer (CRC) is frequently diagnosed at advanced stages owing to the insidious nature of its early clinical manifestations and the lack of validated screening modalities. This diagnostic delay presents a substantial therapeutic challenge, underscoring the need to develop population-based screening strategies. Based on laboratory data, this study constructed a diagnostic model for CRC to explore a screening strategy for population-based tumor census. Methods:In this retrospective case-control study, we analyzed anonymized laboratory parameters and baseline demographic and clinical characteristics of patients with histologically confirmed CRC and age-matched healthy controls undergoing routine health checkups at our tertiary care center between January 2021 and December 2022. Non-parametric comparisons between groups were conducted using the Mann-Whitney U test. Receiver operating characteristic (ROC) curve analysis was performed to evaluate the diagnostic performance of candidate biomarkers. Following variable selection via least absolute shrinkage and selection operator (LASSO), the dataset was partitioned into training and internal validation sets via stratified randomization at a 7:3 ratio. Multiple machine learning algorithms were evaluated, and the optimal predictive model for this study was selected based on this comparative assessment. Subsequently, the influence of individual features on the model predictions was analyzed using SHapley Additive exPlanations (SHAP) values. Results:After rigorous screening, this study included 1,068 patients with histologically confirmed CRC and 1,068 age- and sex-matched healthy controls. Comparative analysis using the Mann-Whitney U test revealed statistically significant intergroup differences (P<0.05) in age and serum biomarkers. ROC curve analysis identified four predictors with superior diagnostic performances for CRC: total protein (TP), carcinoembryonic antigen (CEA), age, and apolipoprotein A1 (ApoA1). LASSO regression selected nine clinically relevant predictors, and various machine learning models were used to classify the data samples. Based on a comprehensive judgment, the Random Forest model [area under the curve (AUC) =0.946, Brier score =0.101] was the best model for the diagnosis of CRC in this study. Conclusions:An effective risk stratification model for CRC screening was developed using routine clinical laboratory parameters and the readily available demographic variable of age. However, being derived from a single-center retrospective cohort, the model requires further validation in multi-center, prospective studies to confirm its generalizability and mitigate potential overfitting.