Pancreatic cancer exhibits a high mortality rate globally, making precise diagnosis particularly critical. Integrin αvβ6 is a promising target for pancreatic cancer imaging, however, high gastrointestinal uptake has been a significant challenge in the clinical translation of previously developed αvβ6-targeted PET probes. In this study, we produced and purified high-quality 64Cu using a medical cyclotron, and developed a 64Cu-labeled αvβ6-targeted molecular imaging probe. Target specificity and binding affinity were evaluated through cell uptake, blocking, and competitive binding assays. Leveraging the relatively long half-life of 64Cu, multi-time-point Micro-PET/CT imaging was performed to explore the feasibility of obtaining high tumor-to-background ratios with low gastrointestinal uptake at extended time points. 64Cu-αvβ6 exhibits high affinity for integrin αvβ6 both in vitro and in vivo. Micro-PET/CT imaging in BxPC-3 (αvβ6 +) tumor-bearing nude mice showed good tumor visualization at all time points, with radioactivity uptake still observed at 24 h. Gastrointestinal uptake was significantly reduced after 8 h, yielding high-quality images with favorable tumor-to-background ratios.

Background: Accumulating evidence substantiates the capacity of targeted radionuclide therapy (TRT) to potentiate tumor immunotherapy responses. However, combining TRT and immunotherapies exhibits substantial varying efficacy, underscoring the importance of investigating mechanisms through which TRT remodels the tumor microenvironment (TME). Despite recent advancements, the specific signaling molecules mediated by TRT that regulate the functions of cytotoxic T lymphocytes (CTLs) remain unclear. Methods: Given the demonstrated potential of α-radionuclides targeting the fibroblast activation protein (FAP) in TRT, this study employed [225Ac]Ac-FAPI-04 to investigate the mechanisms of TRT-induced TME remodeling. The cytotoxic effects were first evaluated in the B16F10 cell line, followed by an assessment of its in vivo therapeutic efficacy in tumor-bearing mice. RNA-sequencing analysis was utilized to elucidate underlying signaling pathways involving immune responses. Results: [225Ac]Ac-FAPI-04 significantly induced DNA double-strand breaks, further elevated ROS levels, and apoptosis in tumor cells. Transcriptomic profiling revealed that [225Ac]Ac-FAPI-04 activates the IL-27/JAK-STAT pathway to amplify interferon-mediated responses, indicating IL-27 serves as a critical regulatory signal for CTLs during TRT. A concomitant observation was the downregulation of Tnfaip3 following [225Ac]Ac-FAPI-04 treatment. Tnfaip3 is implicated in modulating CTLs' behavior and intratumoral infiltration, while its downregulation was associated with the upregulation of cytotoxic effector molecules, as confirmed in this study. Conclusion: This study identifies a TRT-induced pattern of synergistic signaling mediated by IL-27 and Tnfaip3 , which collectively enhances CTL recruitment and potentiates their cytotoxic function. These findings provide a detailed mechanistic rationale for strategies aimed at augmenting the therapeutic efficacy of TRT and immunotherapy combinations. ### Competing Interest Statement The authors have declared no competing interest. National Natural Science Foundation of China, 82372002, 22507148, 82502399 Nonprofit Central Research Institute Fund of the Chinese Academy of Medical Sciences CAMS Innovation Fund for Medical Sciences, 2024-12M-ZH-009, 2021-I2M-3-001, 2023-I2M-2-006, 2021-I2M-1-026, 2025-I2M-XHJC-026, 2025-I2M-XHXX-098 Fundamental Research Funds for the Central Universities, Peking Union Medical College, 3332025193, 3332025067, 3332025068, 3332025153 China Postdoctoral Science Foundation, 2025M773592, 2025M781897 Postdoctoral Fellowship Program of CPSF, GZB20250842 China National Nuclear Corporation Young Talent Program Medical + X Innovation Team of the Discipline Construction Enhancement Project Second Affiliated Hospital of Soochow University, XKTJ-TD202410 Beijing Natural Science Foundation, L248087, L246051, L234044, 7252206

Accumulating evidence substantiates the capacity of targeted radionuclide therapy (TRT) to potentiate tumor immunotherapy responses. However, combining TRT and immunotherapies exhibits substantial varying efficacy, underscoring the importance of investigating mechanisms through which TRT remodels the tumor microenvironment (TME). Despite recent advancements, the specific signaling molecules mediated by TRT that regulate the functions of cytotoxic T lymphocytes (CTLs) remain unclear. Given the demonstrated potential of α-radionuclides targeting the fibroblast activation protein (FAP) in TRT, this study employed [ 225 Ac]Ac-FAPI-04 to investigate the mechanisms of TRT-induced TME remodeling. The cytotoxic effects were first evaluated in the B16F10 cell line, followed by an assessment of its in vivo therapeutic efficacy in tumor-bearing mice. RNA-sequencing analysis was utilized to elucidate underlying signaling pathways involving immune responses. [ 225 Ac]Ac-FAPI-04 significantly induced DNA double-strand breaks, further elevated ROS levels, and apoptosis in tumor cells. Transcriptomic profiling revealed that [ 225 Ac]Ac-FAPI-04 activates the IL-27/JAK-STAT pathway to amplify interferon-mediated responses, indicating IL-27 serves as a critical regulatory signal for CTLs during TRT. A concomitant observation was the downregulation of Tnfaip3 following [ 225 Ac]Ac-FAPI-04 treatment. Tnfaip3 is implicated in modulating CTLs’ behavior and intratumoral infiltration, while its downregulation was associated with the upregulation of cytotoxic effector molecules, as confirmed in this study. This study identifies a TRT-induced pattern of synergistic signaling mediated by IL-27 and Tnfaip3 , which collectively enhances CTL recruitment and potentiates their cytotoxic function. These findings provide a detailed mechanistic rationale for strategies aimed at augmenting the therapeutic efficacy of TRT and immunotherapy combinations.

Metastatic castration-resistant prostate cancer (mCRPC) remains associated with poor long-term outcomes, and survival under current treatment is modest. Prostate-specific membrane antigen (PSMA)-targeted alpha therapy using 225Ac exhibits superior cytotoxicity compared with beta-emitting radiopharmaceuticals; however, dose-limiting xerostomia from salivary gland accumulation restricts clinical application. 225Ac-PSMA-CY313 is a next-generation radioligand designed to reduce off-target salivary uptake while maintaining tumor-targeting efficacy. We evaluated the safety and preliminary efficacy of 225Ac-PSMA-CY313 in heavily pretreated mCRPC patients. This prospective, open-label, single-arm study enrolled 16 patients with progressive mCRPC refractory to standard therapies. Patients received 225Ac-PSMA-CY313 (200 μCi intravenously every 8 weeks). The primary endpoint was treatment-related toxicity per NCI-CTCAE v5.0. Secondary endpoints included ≥ 50 https://www.chictr.org.cn .

Abstract Prostate-specific membrane antigen (PSMA) PET imaging is widely used for diagnosing and staging prostate cancer, yet the currently approved 68Ga- and 18F-labeled tracers are limited by short half-lives and high urinary excretion, which can obscure small pelvic lesions. To overcome these challenges, 64Cu-RAX301 was developed as a high-affinity PSMA PET tracer designed for improved biochemical stability and extended imaging flexibility. The RAX301 precursor and its natCu-RAX301 analog were synthesized with high purity, and radiolabeling with 64Cu was optimized to achieve high molar activity (≥2000 mCi/μmol) and radiochemical purity (>95%), and remaining stable for over 48 hours post-formulation. Surface plasmon resonance revealed sub-picomolar binding affinity (Kd < 1 pM) for both the precursor and natCu-RAX301, markedly stronger than PSMA-617 (22 pM). Consistent with this, cell-based assays showed two-fold higher affinity, 1.5-fold greater uptake, and four-fold higher internalization compared with 64Cu-PSMA-617 (≈80% vs. ≈20%). In LNCaP xenograft mice, 64Cu-RAX301 demonstrated intense and persistent tumor uptake, with tumor-to-muscle ratios exceeding one hundred at 4 hours post-injection. PET imaging in Macaca fascicularis revealed primarily renal clearance with minimal uptake in other organs. In a first-in-human study of ten patients with metastatic castration-resistant prostate cancer (mCRPC), sequential PET/CT scans were performed first with 68Ga-PSMA-11 (5 mCi; imaging at 1 hour post-injection) and then with 64Cu-RAX301 (5 mCi; imaging at 4 and 24 hours post-injection), separated by 1-7 days. 64Cu-RAX301 detected all lesions identified by 68Ga-PSMA-11 while providing higher lesion uptake and superior tumor-to-background contrast. Notably, 64Cu-RAX301 revealed numerous additional small lesions at various sites, including pelvic lymph node metastases that were frequently missed by 68Ga-PSMA-11 due to early bladder activity. The tracer exhibited an excellent safety profile with no significant adverse events. Collectively, these preclinical and early clinical findings demonstrate that 64Cu-RAX301 possesses ultra-high PSMA affinity, favorable pharmacokinetics, and enhanced lesion visualization compared with 68Ga-PSMA-11, supporting its potential as a next-generation PSMA PET imaging agent with improved sensitivity and diagnostic performance in metastatic prostate cancer. Ongoing clinical studies aim to further define its diagnostic impact and broader clinical utility. Citation Format: Fei Chen, Min Hong, Xupeng Hu, Yang Cao, Jie Li, Zhao Li, Shuanglong Liu, Guangzhou Han, Suping Li, Gang Chen. Development and clinical evaluation of 64Cu-RAX301: A next-generation PSMA PET tracer for enhanced detection of metastatic prostate cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2026; Part 1 (Regular Abstracts); 2026 Apr 17-22; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2026;86(7 Suppl):Abstract nr 2610.

This study aimed to establish a fully automated radiosynthesis procedure for [18F] AlF-NOTA-FAPI-04 using the CFN200 multifunctional module, optimize key reaction parameters to improve labeling efficiency, and preliminarily validate its in vivo biological affinity and quality via a translational proof-of-concept PET/CT study. Reaction parameters including precursor amount, AlCl₃ volume, reaction temperature and reaction time were sequentially evaluated and optimized. Quality control including appearance, pH, radiochemical purity, radionuclidic identity, activity, bacterial endotoxins, sterility, ethanol content and residual acetonitrile were performed. Preliminary PET/CT imaging was conducted in a patient with pancreatic cancer for comparison with [18F] FDG. The study was registered with the Chinese Clinical Trial Registry (ChiCTR2300077391). The total synthesis time was approximately 25 min, yielding a decay-corrected radiochemical yield of 29.38 ± 3.3

Copper-64(T1/2 = 12.701 h) is a useful radioisotope for positron emission tomography (PET). This study explores its efficient preparation process through FLUKA simulation calculations combined with the actual production of a 10 MeV low-energy cyclotron. Using 64Ni as the electroplating raw material to prepare the target, a 30 μA proton beam currents was used to bombard for 4–12 h, and 64Cu was obtained through radiochemical separation with an anion-exchange column. The quality control results show that the production capacity of 64Cu is 7.92–34.3 GBq, the radionuclidic purity is > 99.9

This study aimed to develop and evaluate [Cu-64]Cu-PSMA-Q as a novel positron emission tomography (PET) imaging agent for prostate cancer detection, assessing its diagnostic accuracy and clinical applicability in comparison to [F-18]FDG PET imaging. [Cu-64]Cu-PSMA-Q was synthesized, purified, and subjected to comprehensive quality control. Its binding affinity, cellular uptake, and internalization were assessed in vitro using prostate-specific membrane antigen (PSMA)-positive LNCaP C4-2B cells. In vivo toxicity studies were conducted in 12 mouse models (6 per group). Small-animal PET/CT (positron emission tomography/computed tomography) imaging and biodistribution studies were performed on tumor-bearing mice. Clinical evaluation involved PET/CT imaging with [Cu-64]Cu-PSMA-Q in 29 prostate cancer patients, with comparative analysis against [F-18]FDG PET/CT imaging. Radiation dosimetry was calculated using OLINDA/EXM software, and diagnostic performance metrics, including maximum standardized uptake value (SUVmax), mean standardized uptake value (SUVmean), and tumor-to-background ratio, were analyzed using SPSS v24.0, with P < 0.05 considered statistically significant. Comparative analyses utilized t-tests or Mann-Whitney U tests as appropriate. [Cu-64]Cu-PSMA-Q achieved over 99% radiochemical purity and a specific activity of 20.5 +/- 1 GBq/mu mol. In vitro studies demonstrated a dissociation constant (Kd) of 4.083 nM, along with high cellular uptake and internalization in LNCaP C4-2B cells. No significant toxicity was observed in mouse models. Small -animal PET/CT imaging revealed peak tumor uptake at 4 h post-injection in LNCaP C4-2B tumor xenografts. In clinical evaluations, [Cu-64]Cu-PSMA-Q PET/CT detected more lesions than [F-18]FDG, with significantly higher SUVmax, SUVmean, and tumor-to-background ratios. The mean effective radiation dose was calculated as 4.48 +/- 0.99 mSv. [Cu-64]Cu-PSMA-Q PET/CT demonstrated superior lesion detection and higher tumor-to-background ratios compared to [F-18]FDG PET/CT for prostate cancer visualization. Its advantageous properties, including a favorable half-life, excellent safety profile, and enhanced diagnostic accuracy, support its potential for broad clinical adoption. This study establishes a foundation for further validation of [Cu-64]Cu-PSMA-Q in prostate cancer management.

The development of ideal peptide-based radiopharmaceuticals faces critical bottlenecks, primarily due to limited cellular internalization and insufficient deep tissue penetration of peptide carriers. To address this, we developed an intracellular targeting and DNA-adjacent radiotherapeutic strategy using stapled peptides. The MDM2/MDMX-targeting stapled peptide-based radiopharmaceuticals, denoted as [64Cu]Cu-DOTA-STP, exhibited prolonged circulation in the bloodstream and slow systemic clearance. Furthermore, in vitro studies demonstrated that nearly 50% of administered [64Cu]Cu-DOTA-STP was internalized, achieving efficient intracellular accumulation. In addition, [64Cu]Cu-DOTA-STP demonstrated high tumor accumulation, with a standard uptake value of up to 9.39 ± 1.52%ID/g. Finally, targeted radionuclide therapy confirmed that [64Cu]Cu-DOTA-STP effectively inhibited tumor growth, irrespective of p53 phenotypes. Taken together, this study leveraged PET imaging as a noninvasive and longitudinal tool to elucidate the in vivo fate of stapled peptides and demonstrated that stapled peptides can serve as ideal vehicles for developing intracellular protein-targeting radiopharmaceuticals, achieving efficient 64Cu-based targeted radionuclide therapy.

Herein, we report a base-promoted, silylborane-mediated strategy for the transformation of fluorinated carbenes derived from fluoroalkyl ketones, enabling the efficient and stereoselective synthesis of diverse fluorinated 1,3-dienes and alkenes. This metal-free protocol exhibits broad functional group tolerance, accommodating halogens, heteroarenes, and various perfluoroalkyl groups, providing streamlined access to fluorinated frameworks that are otherwise challenging to obtain using conventional methods. Mechanistic studies support a silyl radical-initiated pathway, wherein t-BuONa promotes the in-situ generation of silyl radicals that abstract allylic C-H bonds, triggering downstream selective defluorination. This work establishes a practical platform for the synthesis of valuable fluorinated building blocks and offers insights into silyl radical-driven transformations under metal-free conditions.

This study aims to investigate the diagnostic value of 18F-NaF micro PET/CT imaging in mouse models of acute gouty arthritis (AGA). Three male Balb/c mice were designated as the normal control group (Group A), and 18 male Balb/c mice were used to establish the AGA model (Group B). Group A and model groups B (B 1h, B 3h, B 6h, B 8h, B 12h, B 24h) underwent micro PET/CT imaging 40 minutes after injection of the radiotracer. All groups of mice underwent complete blood count, blood uric acid testing, and pathological biopsy of the ankle joint. The results showed that the counts of inflammatory cells in the blood routine of Group B were higher than those of Group A, and there were statistically significant differences between Group B 6h and B 8h compared to Group A ( P < 0.05). 18F-NaF micro PET/CT imaging revealed abnormal tracer accumulation in the right ankle joints of group B, but no bone destruction were observed on CT at the lesion sites; In group A, there was no obvious abnormal gathering of tracer in the left ankle joint. The ratios of maximum standardized uptake value (SUVmax) of the right and left ankle joints (R/L SUVmax) in Group B were higher than those in Group A, and the difference between Group B 6h and Group A was statistically significant ( P < 0.05). The R/L SUVmax ratios were positively correlated with the counts of white blood cells and neutrophils in the blood routine and microscopic inflammatory cells ( R = 0.79, P < 0.01; R = 0.72, P < 0.01; R = 0.79, P < 0.01, respectively). Overall, 18F-NaF micro PET/CT imaging can detect early bone metabolism changes in AGA and visually monitor its dynamic pathophysiological progression.

[64Cu]Cu-NOTA-JR11 was successfully prepared using a low-energy medical cyclotron and a fully automated solid target purification and synthesis platform. The tracer exhibited excellent radiochemical purity, in vitro stability, and PET imaging properties in preliminary animal studies. The method supports scalable production of 64Cu-labeled SSTR antagonists and provides a basis for further preclinical development and clinical translation.

Mucin 17 (MUC17), a transmembrane mucin, is overexpressed in pancreatic cancer and is associated with tumor proliferation and metastasis. CD3 is an indispensable molecule on the surface of T lymphocytes, which is associated with T cell activation and participates in immune responses. Here, we developed a bispecific T-cell engager radiotracer, 89Zr-M17C3, targeting MUC17 and CD3, to enable noninvasive PET imaging of both tumor cells and T-cell infiltration in pancreatic cancer. 89Zr-M17C3 was synthesized by conjugating AMG199 with zirconium-89 and verified for its radiochemical purity and in vitro stability. The 89Zr-M17C3 probe demonstrated excellent radiochemical purity (>99%) and stability (maintained ≥99% over 120 h). Cellular uptake assays and binding affinity studies were conducted to evaluate the probe's specificity for MUC17 and CD3. Micro-PET/CT imaging and biodistribution studies were performed in MUC17-expressing nude mice and CD3 humanized mice to assess probe uptake in tumors and T-cell-infiltrated tissues. In MUC17-expressing AsPC-1 tumors, probe uptake was significantly higher than in MUC17-negative PANC-1 tumors (SUVmax: 2.26 ± 0.18 vs 1.13 ± 0.14, P < 0.001) and was confirmed to be MUC17-dependent through blocking studies. In CD3 humanized mice, the probe was able to visualize both T-cell infiltration and MUC17-positive tumors, with peak uptake in AsPC-1 tumors (SUVmax: 2.35 ± 0.46) and spleen (SUVmax: 2.19 ± 0.40) at 216 h. Immunohistochemical analysis confirmed the spatial correlation between MUC17 expression and CD3-positive T-cell infiltration in AsPC-1 tumors but not in PANC-1 tumors. In summary, the 89Zr-M17C3 radiotracer exhibited high affinity for MUC17 and CD3 and successfully differentiated MUC17-positive tumors from MUC17-negative tumors while simultaneously providing insight into the T-cell distribution. This study highlights the potential of 89Zr-M17C3 as a versatile imaging tool to support patient stratification and therapeutic monitoring in tumor-targeted immunotherapy, particularly for bispecific T-cell engager-based approaches such as AMG199.

Actinium-225 (²²⁵Ac)-labeled prostate-specific membrane antigen (PSMA) radiopharmaceuticals represent a promising therapeutic approach for metastatic castration-resistant prostate cancer (mCRPC), yet clinical implementation remains limited by the absence of accurate dosimetric assessment methods. The complex decay chain and non-imaging alpha emissions of ²²⁵Ac pose substantial challenges for quantitative imaging. We aimed to evaluate the feasibility of quantitative single photon emission computed tomography (SPECT)-based dosimetry for ²²⁵Ac-PSMA-CY313 therapy by exploiting gamma emissions from daughter radionuclides francium-221 (²²¹Fr) and bismuth-213 (²¹³Bi). Four mCRPC patients received 185.8 ± 11.7 µCi ²²⁵Ac-PSMA-CY313 and underwent multi-timepoint SPECT/CT and whole-body planar imaging at 6, 24, 48, and 96 h post-injection. Quantitative SPECT reconstruction used ordered-subsets expectation-maximization with comprehensive corrections for attenuation, scatter, resolution blur, and crosstalk. Volume of interest were defined using co-registered ¹⁸F-PSMA-CY313 positron emission tomography /computed tomography (PET/CT). Time-activity curves were fitted with mono- or bi-exponential models, and absorbed doses were calculated using validated Monte Carlo-based software and International Commission on Radiological Protection reference phantoms. High-quality quantitative imaging was successfully achieved across all timepoints. Among normal organs, kidneys and liver exhibited the highest absorbed doses (1.55 ± 0.38 Gy and 1.07 ± 0.19 Gy, respectively), corresponding to dose coefficients of 0.23 ± 0.07 Gy/MBq and 0.16 ± 0.03 Gy/MBq. Soft-tissue lesions exhibited higher absorbed doses than bone metastases (5.03 ± 5.51 Gy versus 1.61 ± 2.28 Gy), with corresponding dose coefficients of 0.73 ± 0.80 Gy/MBq and 0.25 ± 0.33 Gy/MBq. Tumor-to-critical organ dose ratios indicated favorable therapeutic windows, with red marrow showing the highest ratio (14.84), followed by adrenal glands (6.35) and salivary glands (4.96), while the dose-limiting kidneys demonstrated a ratio of 1.55. Quantitative SPECT-based dosimetry for ²²⁵Ac-PSMA-CY313 therapy is clinically feasible using standard imaging systems. This methodology demonstrates preferential tumor targeting with acceptable organ-at-risk dose distributions, supporting the therapeutic potential of ²²⁵Ac-PSMA-CY313 for mCRPC and providing a practical framework for personalized dosimetry in targeted alpha therapy.

Background/aim: Papillary thyroid carcinoma (PTC) is the most common form of thyroid cancer. The critical importance of circular RNA (circRNA) in a range of cancer types has been lately recognized. However, research on the functions of circRNAs in PTC has been limited thus far. Therefore, this research aimed at exploring the function and mechanism of circ-methyltransferase-like 15 (METTL15) in PTC cells. Materials and methods: Quantitative measurements of circ-METTL15, miR-200c-3p, and X-linked inhibitor of apoptosis protein (XIAP) in PTC cells were conducted using reverse transcription-quantitative polymerase chain reaction or Western blot analysis. To investigate cell growth, cell counting kit-8 and colony formation tests were employed, apoptosis was analyzed using flow cytometry, and migration and invasion were studied through Transwell assays. The targeted binding sites between miR-200c-3p and circ-METTL15 or XIAP were predicted by starBase and then verified by dual luciferase reporter assay. Results: circ-METTL15 and XIAP were upregulated in the PTC cells, while miR-200c-3p was downregulated. Downregulating circMETTL15 or upregulating miR-200c-3p resulted in inhibited proliferation, migration, and invasion of PTC cells, while promoting apoptosis. miR-200c-3p was the downstream molecule of circ-METTL15, and XIAP was the direct target of miR-200c-3p. Forcing XIAP expression obstructed circ-METTL15 silencing to inhibit PTC cell activity. Conclusion: By coopting miR-200c-3p/XIAP, Circ-METTL15 stimulates aggressive behavior in PTC cells.

BACKGROUND:Noninvasively and accurately predicting subcarinal lymph node metastasis (SLNM) for patients with non-small cell lung cancer (NSCLC) remains challenging. This study was designed to develop and validate a tumor and subcarinal lymph nodes (tumor-SLNs) dual-region computed tomography (CT) radiomics model for predicting SLNM in NSCLC. METHODS:This retrospective study included NSCLC patients who underwent lung resection and SLNs dissection between January 2017 and December 2020. The radiomic features of the tumor and SLNs were extracted from preoperative CT, respectively. Ninety machine learning (ML) models were developed based on tumor region, SLNs region, and tumor-SLNs dual-region. The model performance was assessed by the area under the curve (AUC) and validated internally by fivefold cross-validation. RESULTS:In total, 202 patients were included in this study. ML models based on dual-region radiomics showed good performance for SLNM prediction, with a median AUC of 0.794 (range, 0.686-0.880), which was superior to those of models based on tumor region (median AUC, 0.746; range, 0.630-0.811) and SLNs region (median AUC, 0.700; range, 0.610-0.842). The ML model, which is developed by using the naive Bayes algorithm and dual-region features, had the highest AUC of 0.880 (range of cross-validation, 0.825-0.937) among all ML models. The optimal logistic regression model was inferior to the optimal ML model for predicting SLNM, with an AUC of 0.727. CONCLUSIONS:The CT radiomics showed the potential for accurately predicting SLNM in NSCLC patients. The ML model with dual-region radiomic features has better performance than the logistic regression or single-region models.

Abstract Background Pancreatic cancer-associated fibroblasts (CAFs) play a crucial role in tumor progression and immune evasion. Asperuloside (ASP) is an iridoid glycoside with potential anti-tumor properties. This study aimed to explore the molecular mechanisms of ASP on CAFs, particularly focusing on its effects on activating transcription factor 6 (ATF6), a key regulator of endoplasmic reticulum stress. Method CAFs were treated with different concentrations of ASP (0, 1, 3, and 5 mM), and the role of ATF6 was investigated by over-expressing it in CAFs. Subsequently, western blot was used to detect ATF6, α-smooth muscle actin (α-SMA), fibroblast activating protein (FAP), and vimentin protein levels in CAFs. The collagen gel contraction assay and Transwell assay were applied to evaluate the contraction and migration ability of CAFs. In addition, the interleukin (IL)-6, C–C motif chemokine ligand (CCL)-2, and C-X-C motif chemokine ligand (CXCL)-10 levels were detected by reverse transcription-quantitative polymerase chain reaction (RT-qPCR). Results CAFs had significantly higher expression levels of α-SMA, FAP, and vimentin compared to normal fibroblasts (NFs). ASP significantly inhibited the activation, contraction, and migration of CAFs in a concentration-dependent manner. ASP treatment also reduced the expression of cytokines (IL-6, CCL2, and CXCL10) and down-regulated ATF6 levels. Over-expression of ATF6 mitigated the inhibitory effects of ASP. Conclusion ASP exerts its anti-tumor effects by down-regulating ATF6, thereby inhibiting the activation and function of pancreatic CAFs. These findings suggest that ASP could be a promising therapeutic agent for pancreatic cancer by modulating the tumor microenvironment.

The abnormal activation of the nuclear factor-kappa B (NF-κB)/nod-like receptor family-pyrin domain-containing 3 (NLRP3) signaling pathway is closely related to early brain injury after subarachnoid hemorrhage (SAH). Targeting the NLRP3-inflammasome has been considered an efficient therapy for the local inflammatory response after SAH. Tanshinone IIA (Tan IIA), a major component extracted from Salvia miltiorrhiza, has been reported to have anti-inflammatory effects. The aim of this study was to investigate the effect and mechanism of Tan IIA on early brain injury after SAH. In vivo SAH injury was established by endovascular perforation technique in Sprague-Dawley rats. Limb-placement test and corner turning test were used to measure the behavior. Terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling (TUNEL) staining, hematoxylin-eosin (H&E) staining, and immunofluorescence were used to evaluate the nerve damage. Real-time RT quantitative PCR (RT-qPCR) was used to quantify the levels of inflammatory factors. Western blot was performed for the activation of the NF-κB/NLRP3 pathway. An in vitro SAH model was used to validate the conclusion. We found that the neurobehavioral impairment and cerebral edema in SAH model rats given Tan IIA were alleviated. Further study demonstrated that Tan IIA could inhibit SAH-secondary neuronal apoptosis around hematoma and alleviate brain injury. Tan IIA down-regulated the expression of interleukin-6 (IL)-6, monocyte chemoattractant protein-1 (MCP-1), and tumor necrosis factor (TNF)-α, and inhibited the activation of NF-κB. And the overexpression of pro-inflammatory factors NLRP3, IL-1β, and IL-18 induced after SAH was also reversed by Tan IIA. In conclusions, Tan IIA could inhibit the NF-κB/NLRP3 inflammasome activation to protect and ameliorate SAH-followed early brain injury, and may be a preventive and therapeutic strategy against SAH.