Search Results (119)

Osteosarcoma is a common malignant bone tissue tumour that frequently occurs in the adolescent population, with recurrence and metastasis rates that remain high under current treatment strategies, necessitating the development of cost-effective and efficient anti-osteosarcoma drugs. Sanguinarine, a benzophenanthridine alkaloid derived from plants in the Papaveraceae family, possesses anti-tumour capabilities; however, its clinical application is limited due to poor water solubility and potential organ toxicity. Through screening 30 sanguinarine analogues, in this study we found that analogue 25 exhibited significant activity and could suppress cell proliferation and arrest the cell cycle at the G0/G1 phase in U2OS and MG63 cells, with IC₅₀ values of 1.484 and 1.954 μM, respectively. Analogue 25 could also induce cell apoptosis by increasing the levels of cleaved caspase-9 and BAX. Subsequently, we demonstrated that analogue 25 increased the ROS level, decreased the mitochondrial membrane potential, and increased lipid peroxidation in osteosarcoma cells. After reducing ROS levels through NAC, analogue 25 was able to regulate mitochondrial-mediated osteosarcoma cell apoptosis by modulating the ROS level. Furthermore, analogue 25 could also regulate the expression of ferroptosis-related proteins (including GPX4, SLC7A11 and PTGS2) through the NRF2/GPX4 pathway, ultimately leading to the accumulation of lipid peroxidation and ferroptosis. In a xenograft mouse tumour model derived from osteosarcoma cell lines, analogue 25 exhibited stronger anti-tumour activity than sanguinarine by regulating apoptosis and ferroptosis without causing liver or kidney toxicity. These findings highlight that sanguinarine analogue 25 has beneficial effects in treating osteosarcoma. Full article

Osteosarcoma (OS) is characterized by high metastatic potential and marked chemoresistance, with cancer stem cells (CSCs) serving as major drivers of malignant progression. Canine osteosarcoma (cOS) is considered an ideal comparative medicine model for human osteosarcoma (hOS). Accumulating evidence indicates that aberrant glucose metabolism and hexosamine biosynthetic pathway (HBP, hexosamine biosynthetic pathway)/O-linked N-acetylglucosamine (O-GlcNAc)ylation are involved in tumor progression; however, the precise mechanisms by which they regulate stemness in canine osteosarcoma cells remain unclear. In this study, we comprehensively employed glucose gradient culture, untargeted metabolomics, O-GlcNAc-modified proteomics, in vitro gene silencing, and a subcutaneous xenograft model in nude mice. Cellular functional assays revealed that high glucose significantly enhanced malignant phenotypes and stemness properties of canine osteosarcoma cells. Metabolomic analyses confirmed aberrant activation of the HBP in osteosarcoma cells. Further experiments demonstrated that high glucose enhances HBP flux and O-GlcNAcylation in a dose-dependent manner; silencing of glutamine-fructose-6-phosphate transaminase 1 (GFPT1), O-GlcNAc transferase (OGT), and O-GlcNAcase (OGA) verified that both the HBP pathway and O-GlcNAcylation positively regulate malignant biological behaviors and stemness maintenance. In vivo tumorigenesis assays demonstrated that OGT knockdown markedly suppressed osteosarcoma growth. O-GlcNAc-modified proteomics identified transducin-like enhancer of split 3 (TLE3), nuclear receptor corepressor 1 (NCOR1), and neurogenic locus notch homolog protein 2 (NOTCH2) as key differentially modified proteins, predominantly enriched in the Wingless/Integrated (Wnt) and Notch signaling pathways. Collectively, our findings demonstrate that high glucose activates the HBP pathway, elevates global O-GlcNAcylation levels, and modifies TLE3/NCOR1/NOTCH2, thereby promoting stemness maintenance in canine osteosarcoma stem cells. This study provides novel metabolic targets for precision therapy of osteosarcoma. Full article

Backgrounds: Bladder cancer shows pronounced biological heterogeneity that underlies its variable clinical course and prognosis. Our study aims to delineate clinically relevant differences in bladder cancer using an immunohistochemistry-based molecular subtyping approach. Methods: This retrospective study included 590 patients with bladder cancer treated at a single center. Tumors were stratified into luminal versus non-luminal categories according to CK20, GATA3, CK5/6, and CK14. Associations between molecular subtype, histopathological growth patterns, pathological response to neoadjuvant chemotherapy (NAC), and clinical survival endpoints were analyzed. Overall survival (OS), recurrence-free survival (RFS), and progression-free survival (PFS) were evaluated through Kaplan–Meier survival curves together with Cox proportional hazards regression analyses. Results: Non-luminal tumors exhibited significantly more aggressive pathological growth patterns, including higher levels of tumor budding (p = 0.002), a predominance of non-cohesive or spindle/single-cell architecture (p = 0.003), and more frequent disseminated spreading patterns (p = 0.001), whereas luminal tumors more commonly displayed a higher frequency of tertiary lymphoid structures (TLSs; p = 0.041). Among patients receiving NAC, non-luminal tumors achieved a significantly higher pathological complete response (pCR) rate compared with luminal tumors (p = 0.007), while no significant inter-subtype difference was detected in pathological downstaging between subtypes (p = 0.126). Despite inferior pathological response, luminal tumors demonstrated significantly improved OS (p = 0.003), RFS (p = 0.002) and PFS (p < 0.001) compared with non-luminal tumors. In multivariable Cox regression analysis, molecular subtype was identified as an independent predictor of OS, with luminal tumors showing a lower mortality risk (HR = 0.51, 95% CI 0.33–0.79, p = 0.003). Conclusions: These findings indicate that pathological response and long-term survival follow distinct, subtype-dependent trajectories in bladder cancer. Favorable pathological response does not necessarily correspond to improved long-term survival across molecular subtypes. Full article

Background and Objectives: The HER2-low designation has emerged as a clinically actionable category in breast cancer following the approval of trastuzumab deruxtecan for HER2-low metastatic disease. However, the clinical relevance of HER2-low expression in the neoadjuvant chemotherapy (NAC) setting remains uncertain. This study aimed to evaluate pathological complete response (pCR) as the primary endpoint and overall survival (OS) and disease-free survival (DFS) as exploratory secondary endpoints across HER2-low, HER2-zero, and HER2-positive subgroups in a NAC-treated cohort. Materials and Methods: This single-center retrospective cohort study included 118 patients with histopathologically confirmed invasive breast cancer who received NAC at our institution between January 2010 and March 2021. Patients were classified as HER2-zero (IHC 0, n = 66), HER2-low (IHC 1+ or IHC 2+/FISH-non-amplified, n = 17), or HER2-positive (IHC 3+ or IHC 2+/FISH-amplified, n = 35). pCR was defined as ypT0/Tis ypN0. Univariate analyses used χ² or Fisher’s exact tests; multivariable logistic and Cox regression were performed for adjusted analyses, and Kaplan–Meier survival curves were compared by log-rank, Breslow, and Tarone–Ware tests. Results: The overall pCR rate was 24.6%, differing significantly across HER2 subgroups (HER2-positive 45.7%, HER2-low 29.4%, HER2-zero 12.1%; p = 0.001). After multivariable adjustment for age, ER, PR, and tumor grade, HER2-positive status retained an independent association with pCR (OR 4.37, 95% CI 1.46–13.10, p = 0.008), whereas HER2-low status did not (OR 2.65, 95% CI 0.60–11.75, p = 0.201). At a median follow-up of 48.8 months, neither OS (log-rank p = 0.567) nor DFS (log-rank p = 0.901) differed significantly across HER2 subgroups, and HER2 subgroup status was not independently associated with survival in exploratory Cox models. Conclusions: In this NAC-treated cohort, the unadjusted pCR advantage of HER2-low over HER2-zero tumors was not retained after adjustment for hormone receptor expression and tumor grade, and no HER2 subgroup-specific survival difference was demonstrated. Within the standard NAC framework, HER2-low disease did not show a pCR or survival pattern clearly distinct from HER2-zero disease after adjustment; the small HER2-low subgroup and wide confidence intervals preclude firm conclusions, and an exploratory hormone receptor-stratified analysis indicated that the apparent pooled HER2-low advantage was confined to the hormone receptor-negative subgroup. Future prospective studies powered for hormone receptor-stratified analyses are warranted. Full article

Background: Cold stress is a major abiotic factor limiting rice growth and yield. Elucidating the molecular mechanisms underlying cold tolerance is therefore of great significance for variety improvement. This study focused on the cold-tolerant variety QJ10 and the cold-sensitive variety DHX2, systematically comparing their physiological and transcriptomic differences under cold stress and identifying genes and modules associated with the cold stress response. Methods: After 0, 3, 5, and 7 days of treatment at 4 °C, we measured leaf MDA and Pro contents, as well as SOD and POD activities. We performed multi-time-point transcriptome sequencing using RNA-seq, and conducted data mining and validation through differential expression analysis, Mfuzz trend clustering, WGCNA co-expression network analysis, GENIE3 regulatory network prediction, and qRT-PCR. Results: Compared with DHX2, QJ10 exhibited lower MDA levels and higher levels of Pro, SOD, and POD under cold stress. Transcriptome analysis identified a total of 13,599 differentially expressed genes. Trend clustering revealed that QJ10 primarily maintained genes associated with growth, development, and basal metabolism, whereas DHX2 tended to activate defense responses. WGCNA identified the MEturquoise module, which showed significant positive correlations with both cold treatment duration and the activities of SOD and POD. Genes in this module were significantly enriched in pathways such as carbon metabolism, photosynthesis, and ion transport. Twelve key transcription factors were identified, nine of which were highly expressed at the late stage of cold stress in QJ10. GENIE3 further predicted seven key regulatory factors centered on OsNAC2, OsLBD, and OsARF19; the expression patterns of these factors were validated by qRT-PCR and were consistent with the transcriptomic results. Conclusions: This study revealed that the cold tolerance of QJ10 is associated with enhanced antioxidant capacity, upregulation of genes related to carbon metabolism, and the induced expression of specific transcription factors. The key transcription factors identified here provide candidate genes for studying the molecular mechanisms of cold tolerance in rice. However, their regulatory functions require further experimental validation. Full article

Background and Objectives: Neoadjuvant chemotherapy (NAC) followed by radical cystectomy is the standard of care for eligible patients with locally advanced bladder cancer (LABC). However, adjuvant chemotherapy (AC) remains widely used in real-world practice. Host-related inflammatory and nutritional biomarkers may also influence survival outcomes. This study aimed to compare survival outcomes between NAC and AC and to identify independent prognostic factors for overall survival (OS) and progression-free survival (PFS), with particular emphasis on the Prognostic Nutritional Index (PNI). Methods: This multicenter retrospective study included 262 patients with locally advanced bladder cancer. The median age was 66 years, and 84% of patients were male. Patients were treated with neoadjuvant chemotherapy followed by radical cystectomy or adjuvant chemotherapy after surgery between August 2021 and March 2025. The Prognostic Nutritional Index (PNI) was calculated using pretreatment laboratory values. ROC analysis was used to determine the optimal PNI cut-off for predicting mortality, and the derived threshold (49.97) was applied for stratification in all survival analyses. Survival outcomes were evaluated using the Kaplan–Meier method and compared using the log-rank test. Multivariate Cox proportional hazards regression was used to identify independent prognostic factors. Results: Among 262 patients, 138 (52.7%) received NAC, and 124 (47.3%) received AC. Median follow-up was 33.6 months (95% CI: 29.4–37.8). No statistically significant differences in OS (p = 0.388) or PFS (p = 0.499) were observed between treatment groups. In univariate analyses, nodal stage, pathological complete response (pCR), and PNI were significantly associated with both OS and PFS. In multivariate analysis, low PNI (≤49.97) remained an independent predictor of mortality (HR 1.78, 95% CI 1.04–3.38; p = 0.044), while N3 nodal stage independently predicted disease progression (HR 5.92, 95% CI 1.06–32.84; p = 0.042). Conclusions: In this multicenter real-world cohort, nodal stage and systemic inflammatory-nutritional status were key determinants of prognosis in patients with locally advanced bladder cancer receiving perioperative chemotherapy. PNI emerged as an independent predictor of overall survival, suggesting that host-related biomarkers may improve prognostic stratification beyond traditional clinicopathological factors. Full article

Background: Advanced glycation end products (AGEs) and oxidative stress (OS) have been linked to bone complications related to type 2 diabetes mellitus (T2DM). However, the effects of AGEs and OS on bone marrow mesenchymal stromal cells (BMMSCs), which play a key role in bone homeostasis and repair, remain unclear. Objectives: This study aimed to investigate the effects of AGEs on BMMSCs function and the ability of N-acetylcysteine (NAC) to alleviate AGE-induced OS in a T2DM context. Methods: Bone marrow (BM) and BMMSCs were isolated from Zucker diabetic fatty (ZDF) rats, which serve as a T2DM model, and their lean littermates (ZL, controls) at 24 weeks of age. Results: The results show that long-standing T2DM leads to changes in the BM’s cellular composition and BMMSCs function that are distinct from age-related changes. In vitro, AGEs decreased BMMSCs viability, proliferation, and migration. The effects of AGEs were stronger in BMMSCs derived from a T2DM microenvironment. In both T2DM- and ZL-BMMSCs, AGEs induced cytoplasmic ROS, which was differentially reduced by NAC. The effect of NAC on T2DM-BMMSCs was greater when the cells were pre-treated with NAC 24 h before exposure to AGEs, whereas simultaneous exposure to both resulted in a smaller effect. Conclusions: These results show that AGEs impair BMMSCs expansion and functionality. AGE-induced ROS generation may be a critical factor in this impairment, while NAC was able to reduce OS in BMMSCs from a T2DM context. These findings highlight the vicious negative effects of the T2DM microenvironment on BMMSCs and underscore the need for further studies to better understand the underlying mechanisms and to explore strategies aimed at mitigating OS in the T2DM context. Full article

Jacalin-related lectins play crucial roles in plant adaptation to abiotic and biotic stresses. The rice genome encodes four putative jacalin-related lectin kinase genes (OsJacLKs), but their functions toward environmental stresses remain largely uncharacterized. This study demonstrates that a putative jacalin-related lectin kinase, OsJacLK1, conferred resistance to the rice blast fungus Magnaporthe oryzae rather than salt stress. OsJacLK1 protein exhibited agglutination activities and affinity toward chitin, fungal cell wall, and mannose. OsJacLK1 was transcriptionally activated by stress-related phytohormones salicylic acid (SA), methyl jasmonate (MeJA), abscisic acid (ABA), and indoleacetic acid (IAA), as well as salinity, chitin, and M. oryzae inoculation, suggesting its involvement in broad stress-responsive signaling pathways. Overexpression of OsJacLK1 in rice led to reduced susceptibility to rice blast disease, whereas loss-of-function osjaclk1 lines showed no significant phenotypic difference from wild-type plants upon infection. Enhanced resistance in OsJacLK1-overexpressing lines was associated with a stronger reactive oxygen species (ROS) burst and elevated hydrogen peroxide accumulation, accompanied by the up-regulation of defense-related genes (OsRac1, OsSGT1, OsMAPK6, OsPAL1, OsNAC4, OsPBZ1, OsAOS2, and OsJAZ8). Collectively, our findings establish that OsJacLK1 acts as a positive regulator of rice immunity against M. oryzae, modulating the cellular redox state, highlighting its potential as a candidate for genetic improvement of disease resistance in rice. Full article

Background: Neoadjuvant therapy (NAT) is increasingly investigated in operable pancreatic ductal adenocarcinoma (PDAC), yet its role in strictly resectable disease remains controversial. Randomized trials have been conducted both in borderline resectable and resectable PDAC and have demonstrated survival advantages, while evidence in strictly resectable tumors remains poor. We conducted an umbrella review of systematic reviews and meta-analyses (SRMAs) to comprehensively evaluate the highest level of available evidence on NAT versus upfront surgery in operable PDAC. Methods: We performed an umbrella review of completed SRMAs assessing neoadjuvant chemotherapy (NAC) and/or chemoradiotherapy (NACRT) in resectable and borderline resectable PDAC. MEDLINE/PubMed, Embase, and Cochrane Library were searched from inception through November 2025. Eligible SRMAs reported at least one clinical outcome, including overall survival (OS), disease-free/event-free survival (DFS/EFS), resection rate, R0 resection, nodal status, or perioperative outcomes. Methodological quality was appraised using AMSTAR-2 and ROBIS tools. Overlap among SRMAs was quantified using the Corrected Covered Area (CCA), and RCT-only evidence was prioritized for causal inference. Evidence credibility was graded using an Ioannidis-style classification framework. Results: Thirty-four SRMAs published between 2010 and 2025 were included. In strictly resectable PDAC, RCT-only meta-analyses showed no definitive OS benefit for NAT compared with upfront surgery (pooled HR approximately 0.85, 95% CI 0.68–1.05), although a significant improvement in EFS was observed (HR approximately 0.77, 95% CI 0.65–0.90). Trial sequential analyses suggested insufficient information size for conclusive OS benefit in resectable disease. Conversely, in pooled resectable and borderline resectable populations, NAT significantly improved OS (HR approximately 0.66, 95% CI 0.52–0.85), with subgroup analyses indicating that the survival advantage was primarily driven by borderline resectable tumors. NAT consistently increased R0 resection and node-negative (pN0) rates and reduced non-curative explorations. However, neoadjuvant strategies were associated with treatment-related attrition and, in some analyses, lower overall resection rates. Comparative evidence suggested improved pathological outcomes with chemoradiotherapy versus chemotherapy alone, without a consistent survival advantage. Conclusions: Current high-level evidence supports NAT as the preferred strategy for borderline resectable PDAC, demonstrating consistent survival and pathological benefits. In strictly resectable disease, NAT improves disease-control endpoints and pathological surrogates, but a definitive OS advantage has not been consistently demonstrated in RCT-only syntheses. This should not be interpreted as evidence of equivalence between NAT and a surgery-first strategy, given the heterogeneity, limited power, and therapeutic-era effects of the available literature. Treatment decisions in resectable PDAC should therefore be individualized, balancing potential oncologic benefits against attrition risk. Future adequately powered randomized trials employing contemporary multi-agent regimens are needed to clarify the survival impact of NAT in strictly resectable disease. Full article

Background: Obesity is increasingly prevalent and has been associated with poorer outcomes and higher toxicity in breast cancer treatment. Its impact on tolerance to neoadjuvant chemotherapy (NAC) remains understudied. Objectives: This study aimed to determine the prevalence of overweight and obesity among patients with high-risk early-stage breast cancer (BC) treated with NAC. It evaluated the association between obesity and clinically significant chemotherapy-related toxicities. Pathological complete response (pCR), progression-free survival (PFS), and overall survival (OS) were explored. Methods: This is a retrospective cohort study on stage I–III BC assigned for NAC between January 2020 and December 2022. Patients were categorized by body mass index. Patients with obesity were compared with an age-matched non-obese cohort (1:1). The toxicities evaluated were hospitalization, dose reduction, treatment delay, or treatment discontinuation. Multivariable logistic regression models were performed, adjusting for age, comorbidity burden, and carboplatin use. Results: Of 487 eligible patients, 31.2% were overweight and 27.7% were obese. Peripheral neuropathy was more frequent and severe in obese patients (63.7% vs. 36.3%, p < 0.001). In multivariable analysis, obesity was independently associated with increased odds of global toxicity (OR = 1.83; 95% CI: 1.08–3.15; p = 0.027) and treatment discontinuation (OR = 2.30; 95% CI: 1.18–4.59; p = 0.016). Conclusions: Obesity is highly prevalent among high-risk early BC patients. It is independently associated with increased treatment-related toxicity, particularly discontinuation. These findings highlight the need for closer toxicity monitoring in patients with obesity. Reliance on BMI alone may be insufficient, underscoring the potential value of incorporating body composition assessment into clinical practice. Full article

Cadmium (Cd) toxicity threatens global food security and agricultural sustainability. Transcription factors (TFs) act as master regulators of the complex molecular networks involved in Cd detoxification. This review provides a focused synthesis of the molecular mechanisms governing Cd tolerance in plants, encompassing antioxidant defense, Cd chelation and sequestration, Cd uptake and transport, signal transduction, and damage repair pathways. We highlight the pivotal roles of key TFs in these specific processes, such as OsMYB45 in antioxidant defense, OsIRO2 in regulating chelation and storage, OsNAC5 in modulating Cd transport, and OsE2F in facilitating the repair of DNA and protein damage. Furthermore, we evaluate the potential of harnessing these TF-mediated regulatory mechanisms for developing low-Cd rice varieties. By delineating precise correlations between specific TFs and detoxification pathways, this review proposes actionable molecular strategies to mitigate Cd contamination, thereby contributing to ecological and food safety. Full article

Background/Objectives: Oropharyngeal squamous cell carcinoma (OPSCC) management has shifted following recognition of HPV-driven disease. Neoadjuvant chemotherapy (NAC) has historically failed to improve overall survival (OS) in mixed head and neck cohorts, although contemporary HPV-stratified series suggest NAC may enable treatment de-escalation. We aimed to narratively synthesize OPSCC-specific evidence on NAC focusing on primary and nodal response, pathologic complete response (pCR), survival, and functional outcomes. Methods: We conducted a narrative review of PubMed, selecting primary studies in which OPSCC outcomes were reported separately (surgery- or chemoradiotherapy [CRT]-based strategies; HPV status when available). We extracted study design, treatment regimens, response outcomes, survival, and toxicity data. Results: Pre-HPV studies showed variable responses and no consistent OS advantage over locoregional therapy. In the HPV era, non-comparative cohorts of NAC followed by transoral surgery reported substantial downstaging and high pCR rates at both the primary site and regional nodes, with 3–5-year OS frequently ≥80%. NAC+CRT paradigms demonstrated high clinical CR rates and OS exceeding 80–90%, and lower feeding-tube dependence and reduced swallowing morbidity in de-escalated regimens. Comparative retrospective series suggest NAC + surgery may be associated with lower rates of distant metastases and feeding-tube use compared with CRT or upfront surgery, although interpretation is limited by selection bias, regimen heterogeneity, and small sample sizes. Conclusions: While randomized trials have not established an OS advantage for NAC over standard CRT in head and neck cancer overall, HPV-positive OPSCC shows emerging evidence that systemic intensification with NAC may enable surgical and/or radiation de-escalation with promising oncologic and functional outcomes. Full article

Background: Neuropathic pain is a complex condition often resistant to current therapies due to limited efficacy and adverse effects. Nutraceuticals offer promising alternatives, combining antioxidant and anti-inflammatory properties with good tolerability. This study aimed to compare the effects of a commercial nutraceutical formulation, SUPERALA CARNITINE^® (Pharma Suisse Laboratories SpA, Milan, Italy), containing Alpha-Lipoic Acid (ALA), with a novel formulation, called SUPERALA CARNITINE^® Forte, where ALA and vitamin B6 were replaced by N-acetylcysteine (NAC), Glutathione (GSH), and Uridine monophosphate (UMP). Methods: An indirect gut–peripheral nerve axis was employed to simulate oral absorption, metabolism, and effect on nervous tissues using 3D in vitro models. Both formulations and their individual components were assessed for cytotoxicity and permeability in the gut model (Caco-2 cells in Transwell^®) and, after gut metabolism, for antioxidant capacity, anti-inflammatory activity, and neuroprotective potential in the peripheral nerve model. Results: SUPERALA CARNITINE^® Forte improved cell viability and favoured the maintenance of intestinal integrity, showing enhanced permeability, and significantly reduced oxidative stress (OS) and pro-inflammatory cytokines (TNF-α, IL-2) at the peripheral nervous system. In addition, it increased levels of neuronal markers (p75, MPZ, NRG1, ERβ) and decreased NaV1.7 and NaV1.8 activity, indicating greater neuroprotection and analgesic modulation than the ALA-based formula. Conclusions: The replacement of ALA and vitamin B6 with NAC, GSH, and UMP produced favorable responses in vitro on neuronal cells, supporting a hypothetical potential interest in this nutraceutical combination and justifying further future in vivo investigations. Full article

Background/Objectives: Optimal selection for perioperative therapy in urothelial carcinoma (UC) remains uncertain. We evaluated the efficacy of neoadjuvant and/or adjuvant chemotherapy (NAC/AC) for patients with bladder cancer (BC) and upper tract UC (UTUC), examined the role of lymphovascular invasion (LVI), and considered the implications for adjuvant nivolumab. Methods: We retrospectively analyzed consecutive patients who underwent radical cystectomy or radical nephroureterectomy at a single center (July 1998–April 2021; observation to 31 March 2025). After exclusions, 252 BC and 153 UTUC patients were included. Endpoints were cancer-specific survival, progression-free survival (PFS; BC), non-urinary-tract recurrence-free survival (NUTRFS; UTUC), and overall survival (OS). Survival was estimated by Kaplan–Meier analysis and compared by log-rank tests. Results: For BC, AC did not improve the PFS or OS in the overall pT ≥ 2 population, whereas node-positive (pN+) disease derived significant benefits in both endpoints among NAC-naïve patients (PFS and OS, p = 0.002 and p = 0.008). For UTUC, AC conferred no advantage in NUTRFS or OS for the overall pT ≥ 2 population. However, NUTRFS benefits emerged in the pN+ subset (p = 0.049), although the OS was not improved. Among NAC-treated BC, the outcomes were poorest for ≥ypT3 and ypN+, whereas ypT ≤ 2 fared better. LVI was associated with adverse outcomes and was borderline higher in pN+ versus pT ≥ 2/pN− for BC (p = 0.056) and significantly higher for UTUC (p = 0.012). Conclusions: In this retrospective, single-center cohort, our exploratory analyses suggest that perioperative benefit is largely node-dependent, supporting prioritizing systemic therapy for pN+ disease and cautioning against routine AC for pT2/ypT2 without nodal involvement. After NAC, adjuvant therapy appeared most justified for ≥ypT3/ypN+. Prospective biomarker-integrated validation is warranted and, given the small and underpowered subgroups and the potential for selection and immortal time biases, these observations should be interpreted as hypothesis-generating rather than causal. Full article

The plant NAC (NAM, ATAF1/2, and CUC2) transcription factor family plays an important regulatory role in stress response. In this study, we analyzed the rice transcription factor OsNAC113 and elucidated its tissue-specific characteristics and stress response regulatory mechanisms. qRT-PCR results showed that under laboratory-simulated drought, high salt, temperature stress, and hormone treatments, such as abscisic acid (ABA) and gibberellic acid (GA₃), the expression level of OsNAC113 significantly changed, indicating that OsNAC113 responds to various stress conditions. Targeted creation of the rice (Oryza sativa L. spp. japonica) OsNAC113 (LOC_os08g10080.1) mutant based on the CRISPR-Cas9 genome editing strategy revealed its response to salt stress (200 mM). The growth status and survival rate of the mutant under high-salt stress were significantly higher than those of the wild type. Testing showed that the mutant exhibited increased relative water, chlorophyll, and soluble sugar contents under salt stress than the wild type. The malondialdehyde content in the mutant was lower, and the activities of superoxide dismutase, peroxidase, and catalase were higher than those in the wild type, indicating that the mutant with functional loss caused by knocking out OsNAC113 had a significantly enhanced tolerance to salt treatment. Using RNA-seq to detect genome-wide changes in OsNAC113 mutant materials under stress, KEGG annotation showed that knocking out OsNAC113 resulted in regulatory changes in “plant hormone signaling pathway” and “MAPK signaling pathway,” and GO and KEGG annotations showed significant changes in “amino acid transport and metabolism,” “carbohydrate transport and metabolism,” “lipid transport and metabolism,” and “replication, recombination, and repair.” OsNAC113 may be involved in the response to salt stress by regulating these signaling pathways. Using comparative metabolomic analysis, we further elucidated the function of OsNAC113 in physiological metabolic pathways. The knockout of OsNAC113 resulted in changes in various important metabolic pathways in plants, including flavonoid biosynthesis and ABC transporters. Therefore, it is suggested that OsNAC113 is involved in these metabolic processes and affects their regulation in high-salt environments. These results provide a theoretical foundation and reliable material for the molecular breeding of rice. Full article