Gastric adenocarcinoma (GAC) remains a leading cause of cancer-related mortality, particularly in patients with peritoneal carcinomatosis, for whom effective therapies are limited. We investigated the therapeutic efficacy and molecular mechanism of CYD-4-61, a BAX activator, using human GAC cell lines, patient-derived xenograft models, genetically engineered mouse models, and a syngeneic mouse model. CYD-4-61 potently inhibited tumor cell proliferation, induced apoptosis, and suppressed cancer stem cell-like properties, with enhanced activity in radiation-resistant GAC cells. Mechanistically, CYD-4-61 activated the BAX-caspase pathway, leading to SOX9 protein reduction. Integrated bulk and single-cell transcriptomic analyses identified SOX9-dependent transcriptional programs as major targets of CYD-4-61. Functional rescue experiments together with chromatin immunoprecipitation and CUT&RUN analyses supported CDK4 as a SOX9-regulated gene and demonstrated suppression of the SOX9-CDK4 regulatory axis following CYD-4-61 treatment. In multiple preclinical models, CYD-4-61 significantly inhibited tumor growth and improved the therapeutic response to anti-programmed cell death protein 1 (PD-1) therapy while modulating the tumor immune microenvironment. Clinically, co-expression of SOX9 and CDK4 was associated with diffuse-type GAC and poor patient outcomes. These findings identify the BAX-SOX9-CDK4 axis as an important mechanism contributing to the antitumor activity of CYD-4-61 and provide a strong preclinical rationale for its further development as a therapeutic strategy for aggressive GAC.

Intrapancreatic fat deposition (IPFD) is associated with pancreatic diseases, such as pancreatitis, type 2 diabetes mellitus, and pancreatic cancer, and so on. Although non-invasive imaging has been used to quantify IPFD in clinical settings, this approach does not fully reflect the incidence and prevalence of IPFD, especially its occurrence in a cohort of elderly individuals, which is difficult to determine from routine medical visits. This study aims to systematically evaluate IPFD in elderly individuals, classify its subtypes, and assess their relationship with other pancreatic lesions. In the present study, 85 cadaveric pancreatic specimens (median age 88.0 years) without any known pancreatic diseases or prior abdominal surgery were subjected to histopathological and immunohistochemical analyses as needed. IPFD was classified into three predominant types: fatty infiltration of the pancreas (FIP), fatty replacement of the pancreas (FRP), and irregular intralobular fatty degeneration (IIFD), with 81

BACKGROUND:Silent corticotroph adenomas (SCAs) are an aggressive pituitary neuroendocrine tumor (PitNET) subtype lacking effective medical therapies and showing a high rate of recurrence. The molecular mechanisms driving their proliferation remain poorly understood. Although metabolic reprogramming is a hallmark of cancer, neither lipid metabolism nor its regulatory pathways have been systematically investigated in SCAs. METHODS:We performed integrative analyses of bulk and single-cell RNA sequencing datasets from SCAs, functioning corticotroph adenomas (FCAs), and normal pituitary tissue to characterize GAL expression and associated signaling pathways. Mechanistic studies employed GAL gain- and loss-of-function models, RNA sequencing, luciferase reporter assays, lipidomics, and pharmacological inhibition. Regulation of GAL by MYCN was assessed through promoter transactivation assays. The therapeutic efficacy of SREBP1 inhibition (fatostatin) was evaluated both in vitro and in vivo. RESULTS:We identified a marked downregulation of galanin (GAL) in SCAs and demonstrated that GAL loss promotes tumor cell proliferation. GAL deficiency activated the PI3K-Akt-mTOR signaling cascade, resulting in increased activity of SREBP1, a key transcriptional regulator of lipogenesis. Enhanced fatty-acid synthesis provided metabolic support for SCA growth. We further uncovered a linear regulatory axis in which MYCN directly upregulates GAL; however, MYCN downregulation in SCAs suppresses GAL expression, thereby enabling SREBP1-driven lipogenesis. Pharmacological inhibition of SREBP1 with fatostatin reduced lipogenesis and significantly suppressed SCA growth in vitro and in vivo. CONCLUSIONS:Our findings reveal a previously unrecognized MYCN-GAL-SREBP1 lipogenic axis that drives SCA proliferation. SREBP1-dependent lipogenesis represents a promising and druggable therapeutic vulnerability for the treatment of SCAs.

Dual-specificity phosphatase 5 (DUSP5) is a key regulator of the mitogen-activated protein kinase (MAPK) pathway, with established roles in various types of cancer. However, its function in esophageal squamous cell carcinoma (ESCC) remains unclear. This study combines single-cell transcriptomics with in vitro and in vivo models to investigate the role of DUSP5 in ESCC. Single-cell RNA sequencing revealed tumor-infiltrating myeloid populations, including apolipoprotein C-positive (APOC⁺) macrophages, which interact with tumor cells via the amphiregulin-epidermal growth factor receptor (AREG-EGFR) axis, activating MAPK/extracellular signal-regulated kinase (ERK) signaling to promote tumor growth and immune modulation. We identified a prognostic gene signature linked to these macrophages. DUSP5 expression was downregulated in ESCC tissues, and its overexpression inhibited cell proliferation, induced senescence and apoptosis, and suppressed migration and invasion. In mouse xenografts, overexpression of DUSP5 reduced tumor growth and metastasis. Mechanistically, DUSP5 inhibited ERK1/2 activation, and its tumor-suppressive effects were reversed by ERK1/2 activation. Moreover, ETS Like-1 protein (ELK1), an ERK1/2 downstream transcription factor, was identified as a negative regulator of DUSP5. In a carcinogen-induced model, DUSP5 knockout increased tumor burden, effects reversed by ERK1/2 inhibition. Our findings indicate that the DUSP5-ERK1/2-ELK1 signaling axis, modulated by tumor-infiltrating myeloid cells, contributes to ESCC progression and represents a promising source of biomarkers and therapeutic targets.

M2-like macrophages and CD8+T cells are key immune components that influence tumor behavior and treatment response. Ubiquitin-specific protease 32 (USP32) is established as a key oncogenic factor in gastric cancer (GC). This study aimed to investigate the role of USP32 in regulating M2 macrophage polarization and CD8+T cell dysfunction in GC. Macrophages derived from THP1 cells (THP1-M0) or CD8+T cells were co-cultured with transfected AGS and HGC-27 GC cells. The proportion of CD206+ M2 macrophages and the apoptosis of CD8+T cells were assessed by flow cytometry. Cell invasion was analyzed by transwell assay. The interaction between USP32 and death-associated protein kinase 1 (DAPK1) was verified by GST pull down and Co-immunoprecipitation (Co-IP) experiments. The effect on tumor growth was tested by subcutaneous xenograft studies. USP32 and DAPK1 were overexpressed in GC tissues and cell lines. Mechanistically, USP32 stabilized DAPK1 protein through deubiquitination. DAPK1 downregulation reversed USP32-mediated enhancement in GC cell invasion, macrophage M2 polarization, and CD8+T cell apoptosis in vitro. USP32 depletion exhibited an in vivo anti-growth effect on AGS subcutaneous xenografts. This study identifies the USP32/DAPK1 cascade as a crucial regulator of M2 macrophage polarization and CD8+T cell apoptosis in GC, providing a novel mechanistic link between post-translational regulation and tumor immune evasion.

Tumor innervation (TIN) and perineural invasion (PNI) are well-established pathological features of pancreatic ductal adenocarcinoma (PDAC) that drive its aggressiveness and associated pain. Here, we reveal that regenerating islet-derived (Reg) proteins, secreted by peritumoral exocrine acinar cells, facilitate TIN and PNI through two paracrine mechanisms. In PDAC cells, Reg proteins drive cancer invasiveness along nerves via autocrine transforming growth factor β (TGF-β) signaling. In neurons, Reg proteins are neurotrophic and potentiate neuronal excitability, resulting in hyperinnervation and pain. Interleukin-22, primarily produced by CD4+ T cells, triggers Reg expression. Exostosin-like glycosyltransferase 3 (EXTL3) is the functional receptor for Reg proteins in both cell types. Genetic silencing of Reg or EXTL3 reduces TIN, nerve-cancer proximity, PDAC progression, and pain behavior in mice. Clinically, the Reg-EXTL3-TGF-β axis correlates with increased TIN and PNI severity, poor prognosis, and greater pain. Thus, targeting the Reg-EXTL3 axis may be an attractive strategy for mitigating neural-associated adverse consequences in PDAC.

Administration of selective serotonin reuptake inhibitors (SSRIs) is associated with a reduced cancer risk and shows significant anti-tumor effects across multiple tumor types, suggesting the potential for repurposing SSRIs in cancer therapy. Nonetheless, the specific molecular target and mechanism of action of SSRIs remain to be fully elucidated. Here, we reveal that citalopram exerts an immune-dependent anti-tumor effect in hepatocellular carcinoma (HCC). Interestingly, the anti-HCC effects of citalopram are not reliant on its conventional target, the serotonin transporter. Through various drug repurposing approaches, including global reverse gene expression profiling, drug affinity responsive target stability assay, and molecular docking, the complement component 5a receptor 1 (C5aR1) is identified as a new target of citalopram. C5aR1 is predominantly expressed by tumor-associated macrophages, and citalopram treatment enhances local macrophage phagocytosis and elicits CD8+ T anti-tumor immunity. C5aR1 deficiency or depletion of CD8+ T cells hinders the anti-HCC effects of citalopram. Collectively, our study reveals the immunomodulatory roles of citalopram in inducing anti-tumor immunity and provides a basis for considering the repurposing of SSRIs as promising anticancer agents for HCC treatment.

Metabolic reprogramming is a hallmark of cancer. The“Warburg effect”, also known as aerobic glycolysis, is an essential part of metabolic reprogramming and a central contributor to cancer progression. Moreover, hypoxia is one of the significant features of pancreatic ductal adenocarcinoma (PDAC). Under hypoxic conditions, the “Warburg effect” occurs to meet the nutrient and energy demands of rapid genome replication, remodeling the tumor microenvironment (TME) and influencing tumor immunity. α-Enolase (ENO1) is a multifunctional protein, acting as a glycolytic enzyme that catalyzes the conversion of 2-phosphoglyceric acid to phosphoenolpyruvic acid. ENO1 was found to be overexpressed in multiple types of cancers. Here, we investigated the role of ENO1 in modulating the PDAC microenvironment.Using bioinformatic analyses, we demonstrated that ENO1 was highly expressed in PDAC patients, which was related to a poor prognosis. In vitro, Eno1 knockdown resulted in reduced PDAC cell proliferation and colony formation, along with enhanced apoptosis in PDAC cells. In vivo, tumorigenesis was suppressed in mouse PDAC models by Eno1 knockdown. Flow cytometry analysis revealed that high expression of Eno1 altered the tumor immune microenvironment (TIME), particularly the impaired tumor infiltration and function of CD8+ T cells. Mechanistic studies revealed that ENO1 upregulated PD-L1 to prevent CD8+ T cells infiltration through the hypoxia-inducible factor (HIF)-1α signaling pathway, leading to PDAC progression.In conclusion, our findings indicate that ENO1 might serve as a potential biomarker for PDAC and a novel onco-immunotherapeutic target via its role in altering the TIME.

Pituitary neuroendocrine tumors (PitNETs) are pathologically characterized by dysregulation of neuroendocrine function and systemic disruption of hormonal homeostasis, yet their regulatory effects on peripheral immune networks remain poorly characterized. Here, we systematically analyzed bulk RNA sequencing (RNA‑seq) from 883 PitNET tumors, 108 PitNET‑associated peripheral blood mononuclear cells (PBMC) samples, and 175 healthy PBMC controls, combined with 69 single‑cell RNA sequencing (scRNA-seq) samples covering tumors, normal pituitaries, as well as tumor‑derived and normal PBMCs. We identified a systemic immune disequilibrium in PitNET patients, characterized by increased circulating lymphocyte proportions, accompanied by upregulated cytokine-receptor interaction signatures. Notably, tumor resection reversed this imbalance, as supported by the normalization of monocyte and neutrophil counts, validated by flow cytometry and routine blood data from 600 samples (200 healthy controls and 200 PitNET patients with paired pre- and post-surgery follow‑up). Trajectory analysis identified terminally differentiated, secretory-specialized cell populations with lineage-specific hormone and cytokine hypersecretion. Ligand-receptor inference suggested these tumor-derived factors potentially engage circulating immune cell receptors. A random‑forest classifier based on PBMC transcriptomes distinguished PitNET subtypes, underscoring the diagnostic potential of peripheral immune signatures. Furthermore, in an estrogen-induced rat model, elevated PRL level coincided with the same peripheral immune skewing. Overall, our work provides a valuable resource and demonstrates PitNETs can be systemic immune modulators, where intrinsic hormone secretory activity and monocyte-lymphocyte imbalance collectively drive peripheral immune dysfunction.

Mitochondrial uncouplers dissipate proton gradients and deplete ATP production from oxidative phosphorylation (OXPHOS). While the growth of prostate cancer depends on OXPHOS-generated ATP, the oncogenic pathway mediated by the transcription factor E2F1 is crucial for the progression of this deadly disease. Here, we report that mitochondrial uncouplers, including tizoxanide (TIZ), the active metabolite of the Food and Drug Administration (FDA)-approved anthelmintic nitazoxanide (NTZ), inhibit E2F1-mediated expression of genes involved in cell cycle progression, DNA synthesis, and lipid synthesis. Consequently, NTZ/TIZ induces S-phase kinase-associated protein 2 (SKP2)-mediated G1 arrest while impeding DNA synthesis, lipogenesis, and the growth of prostate cancer cells. The anti-cancer activity of TIZ correlates with its OXPHOS-uncoupling activity. NTZ/TIZ appears to inhibit ATP production, thereby activating the AMP-activated kinase (AMPK)-p38 pathway, leading to cyclin D1 degradation, Rb dephosphorylation, and subsequent E2F1 inhibition. Our results thus connect OXPHOS uncoupling to the inhibition of an essential oncogenic pathway, supporting repositioning NTZ and other mitochondrial uncouplers for prostate cancer therapy.

Survival critically depends on maintaining blood glucose levels to provide essential energy, especially during emergencies such as the fight-or-flight response, when timely glucose control via neural integration is vital. However, pancreatic islets constitute only a small fraction of the pancreas and are dispersed throughout the organ, raising the fundamental question of how the nervous system coordinates synchronized control of multiple islets. Using whole-organ clearing and 3D imaging, we mapped pancreatic sympathetic innervation, revealing specialized anatomical integration between sympathetic nerves and islets. Transplanted islets intrinsically attracted sympathetic nerves independent of their native environment. Chronic islet injury disrupted sympathetic innervation and markedly impaired nerve regeneration after denervation. Sympathetic denervation markedly elevated islet-derived Reg2 and Reg3β; administration of these proteins accelerated sympathetic regeneration and improved islet graft function. Our findings identify an islet-sympathetic architecture actively maintained by islets, uncovering an endocrine-driven mechanism for neural regulation, highlighting Reg2 and Reg3β as therapeutic candidates for diabetes management. ### Competing Interest Statement The authors have declared no competing interest. This study was supported by the National Natural Science Foundation of China, 82230087, 82350123, 82203228 the Shanghai Municipal Education Commission-Gaofeng Clinical Medicine Grant Support, 20181708 Innovative research team of high-level local universities in Shanghai, SHSMU-ZDCX20210802 Shanghai Pilot Program for Basic Research - Shanghai Jiao Tong University, 21TQ1400225 111 project, no. B21024 Shenyang Science and Technology Plan in 2022, 22 - 101 - 0 - 22

BACKGROUND:Pancreatic neuroendocrine microtumors (PNEMTs) are small (< 5 mm), non-functioning, well-differentiated neuroendocrine neoplasms. Although they are rare, they are not invariably benign. PNEMTs are typically discovered incidentally during autopsy. However, data regarding the occurrence of PNEMTs in the elderly population, particularly those identified incidentally in cadaveric studies, remain limited. AIM:To investigate the prevalence and histopathological characteristics of PNEMTs in elderly individuals by analyzing cadaveric pancreatic tissues. METHODS:We conducted a retrospective analysis of 85 pancreatic specimens (age range: 58-109 years) obtained from cadavers for anatomical education and research at the Department of Life Dentistry, Nippon Dental University. Paraffin sections of the pancreatic head, body, and tail were prepared for histological and immunohistochemical analysis. RESULTS:Five cases with PNEMTs (5/85, 5.9%; male, n = 33; female, n = 52; mean age: 85.8 ± 12.1 years) were identified. The tumors were solitary, well circumscribed, and located within the pancreatic parenchyma (body: n = 4; tail: n = 1), and all were < 5 mm (range: 0.54-2.20 mm) in size. All tumors showed strong chromogranin A and synaptophysin positivity, and were predominantly glucagon (GLU)-positive. Ki-67 immunostaining indicated minimal proliferative activity; therefore, these tumors were considered non-functioning, GLU-producing, well-differentiated grade 1 PNEMTs. CONCLUSION:Small, predominantly low-grade, GLU-secreting PNEMTs were present in 5.9% of elderly individuals, highlighting the prevalence of subclinical PNEMTs and the need for careful follow-up.

Lymph nodes (LNs) are critical peripheral immune organs extensively innervated by both sympathetic and sensory nerves. During tumor metastasis, LNs undergo significant structural remodeling and enlargement; however, the role of neural innervation in this process remains unclear. Here, using whole-organ three-dimensional (3D) imaging, we observed pronounced elongation and increased branching specifically in sympathetic nerve fibers, but not sensory nerves, during tumor-induced LN enlargement (TLNE), suggesting adaptive neural remodeling. Single-nucleus RNA sequencing further revealed activation of fibroblastic reticular cells (FRCs) during TLNE, characterized by enriched neuro-related signaling pathways and substantial secretion of hepatocyte growth factor (HGF). Functional validation using targeted HGF inhibitors and adeno-associated virus (AAV)-mediated HGF silencing confirmed that FRC-derived HGF critically drives sympathetic nerve growth. Additionally, both HGF inhibition and sympathetic nerve denervation significantly reduced TLNE and tumor-induced LN metastasis, highlighting the importance of adaptive sympathetic innervation in tumor-associated LN remodeling. These findings identify a previously unrecognized FRC-HGF-sympathetic nerve axis and propose neural regulation as a potential therapeutic strategy for tumor-induced LN metastasis.

Following the publication of the above paper, it was drawn to the Editors' attention by a concerned reader that the western blot data shown in Fig. 2C on p. 5863, the cell adhesion assay data in Fig. 3C on p. 5864, and scratch‑wound assay data shown in Figs. 3D and 4B on p. 5865 were strikingly similar to data appearing in different form in other articles written by different authors at different research institutes that had already been published elsewhere prior to the submission of this paper to Molecular Medicine Reports. In view of the fact that the abovementioned data had already apparently been published previously, the Editor of Molecular Medicine Reports has decided that this paper should be retracted from the Journal. The authors were asked for an explanation to account for these concerns, but the Editorial Office did not receive a reply. The Editor apologizes to the readership for any inconvenience caused. [Molecular Medicine Reports 17: 5860‑5868, 2018; DOI: 10.3892/mmr.2018.8606].

BackgroundPacinian corpuscles (PCs) are pressure- and vibration-sensitive mechanoreceptors found in hairless skin, external genitalia, joints, ligaments, lymph nodes, prostate, bladder, etc. While they are documented in the pancreas of cats, their presence in the normal pancreas remains speculative.PurposeThe present study therefore investigated the distribution of PCs in the normal human pancreas and compared the findings with those in several other animal species.MethodsThe study subjects included 74 human cadaver specimens, 3 Cynictis penicillata, 2 Saguinus mystaxs, 1 Felis domesticus, and 10 Suncus murinus. Pancreatic tissues were prepared as paraffin sections for histological and immunohistochemical analyses of the main constituents of PCs (central axon, inner core, and outer core capsule).ResultsPCs were found in the pancreas of five human cadavers (7%), as well as in one C. penicillata, one S. mystax and one F. domesticus but not in S. murinus. The PCs varied in size, with the largest in the human pancreas measuring up to 1,106 μm—far exceeding those in animal pancreata, but less numerous than those in animals. Morphologically, animal PCs were mainly typical oval shapes, whereas PCs in the human pancreas were mostly irregular in shape. In addition, we found that PCs in animals and human pancreata had similar structures, with consistent expression of protein gene product 9.5, in axonic profiles, and diffuse vimentin immunoreactivity in the inner core, outer core, and capsule.ConclusionThis study confirmed the presence of PCs in a small number of healthy humans and some animal pancreata. The number, distribution characteristics, and morphology of PCs in the pancreata of animals and humans are quite different; however, their structures and immunohistochemical profiles are similar. The presence of PCs in the normal human pancreas is also a mystery, and the physiological role of PCs in the human pancreas requires further clarification.

Lymph node (LN) metastasis of gastric cancer (GC) is one of the important pathways of GC metastasis, indicating the clinical staging and prognosis of patients. To investigate the underlying mechanism during the process of GC-induced LN metastasis, 7 pairs of GC tissues, paracancerous (PC) tissues, GC-positive LN (LN.P) and GC-negative LN (LN.N) tissues from GC patients with homogeneity were selected for RNA sequencing (RNA-seq) analysis. Tensin 4 (TNS4) was screened out and found to be significantly upregulated in LN.P tissues and closely related with the characteristics of GC. In vitro and in vivo experiments demonstrated that knockdown of TNS4 could significantly inhibit LN metastasis of GC cells and activation of fibroblastic reticular cells (FRCs) in LNs, thus inhibiting LN expansion induced by tumor cell invasion. Moreover, TNS4 was found to be interacted with integrin beta 1 (ITGB1) on FRCs, thereby affecting the binding of transforming growth factor β1 (TGF-β1) to ITGB1 and subsequently regulating downstream signaling molecules, and supporting the GC cell-induced LN metastasis.

Inflammatory bowel disease (IBD) commonly coexists with non-alcoholic fatty liver disease (NAFLD). Despite metabolic factors being less involved, IBD patients exhibit a higher risk of developing NAFLD compared to non-IBD individuals. Given the shared role of gut dysbiosis in the pathogenesis of both diseases, this study investigated the involvement of gut microbiota and associated metabolic pathways in IBD-associated NAFLD (COMO). A retrospective analysis of clinical profiles from 490 IBD, 89 NAFLD, and 68 COMO patients was conducted. Fecal samples from 30 IBD, 32 NAFLD, 26 COMO patients and 29 healthy controls were prospectively collected and subjected to 16 S rRNA gene sequencing for microbial community analysis and functional pathway prediction. Subsequently, machine learning modeling was employed for feature importance analysis and identification of COMO patients. Demographic analysis revealed that COMO patients developed NAFLD earlier than NAFLD alone, with fewer metabolic associations with hypertension, hyperlipidemia and glucose dysregulation. Compared with IBD and NAFLD groups, COMO microbiota exhibited lower alpha diversity, with beta diversity aligning with IBD but distinct from NAFLD group. Shared microbial signatures included increased Lactococcus and decreased Coprococcus 3 and Ruminococcus 2, which was correlated with 11 metabolic pathways: five vitamin B pathways (thiamine, vitamin B6, biotin, folate and riboflavin), isoflavonoid, caffeine, phosphonate, cyanoamino acid, lipoic acid and ubiquinone pathways. Integrated microbial-metabolic machine learning models (logistic regression, random forest, support vector machine, and XGBoost) achieved AUC of 0.818–0.864 for COMO identification. Our findings implicate microbiota-mediated metabolic reprogramming in IBD-associated NAFLD pathogenesis, highlighting potential therapeutic targets for the treatment and prevention of NAFLD in IBD.

Objective: To evaluate the efficacy and safety of neoadjuvant chemotherapy (NAC) compared to NAC combined with immune checkpoint inhibitors (ICI) in patients with muscle-invasive bladder cancer (MIBC). Propensity score matching (PSM) was employed to assess the impact of these two treatment regimens on the pathological complete response rate (pCR) and overall survival (OS). Methods: A retrospective analysis was conducted on 320 MIBC patients treated at the Cancer Hospital affiliated to Sun Yat-sen University Gansu Hospital between January 2017 and June 2022. Patients were categorized into the NAC group (n=194) and the NAC+ICI group (n=126) based on their treatment regimens. After PSM, 154 patients were included, with 77 in each group. Baseline characteristics, clinical efficacy, and prognosis were analyzed using various statistical methods. Results: Before PSM, significant differences were observed between the groups in baseline characteristics, including tumor diameter, tumor number, and adjuvant treatment (all P<0.05). After PSM, these differences were no longer statistically significant (all P>0.05). The NAC+ICI group demonstrated a significantly higher pCR rate both before and after PSM (both P<0.001). Similarly, pathological downstaging rates were higher in the NAC+ICI group before and after PSM (both P<0.001). However, there was no significant difference in disease control rates between the two groups before (P=0.057) and after PSM (P=0.240). Logistic regression analysis identified the treatment regimen (before PSM: P<0.001, OR=0.161; after PSM: P<0.001, OR=0.141) and complications (before PSM: P=0.005, OR=2.339; after PSM: P=0.019, OR=2.753) as independent risk factors for pCR. Cox regression analysis revealed that age (before PSM: P<0.001, HR=1.059; after PSM: P=0.011, HR=1.066), pretreatment T stage (before PSM: P<0.001, HR=2.342; after PSM: P<0.001, HR=3.244), tumor diameter (before PSM: P=0.005, HR=1.810; after PSM: P=0.025, HR=2.077), and treatment outcome (before PSM: P<0.001, HR=1.722; after PSM: P=0.020, HR=1.444) were independent prognostic factors for OS. Conclusion: NAC combined with ICI significantly improves pCR and pathological downstaging rates in MIBC patients. Independent prognostic factors affecting OS include age, pretreatment T stage, tumor diameter, and treatment outcome.