Casticin, the flavonoid extracted from Vitex rotundifolia L, exerts various biological effects, including anti-inflammatory and anti-cancer activity. The aim of this study is to investigate the effects and mechanisms of casticin in human gallbladder cancer cells. Human NOZ and SGC996 cells were used to perform the experiments. CCK-8 assay and colony formation assay were performed to evaluate cell viability. Cell cycle analyses and annexin V/PI staining assay for apoptosis were measured using flow cytometry. Western blot analysis was used to evaluate the changes in protein expression, and the effect of casticin treatment in vivo was experimented with xenografted tumors. In this study, we found that casticin significantly inhibited gallbladder cancer cell proliferation in a dose- and time-dependent manner. Casticin also induced G0/G1 arrest and mitochondrial-related apoptosis by upregulating Bax, cleaved caspase-3, cleaved caspase-9 and cleaved poly ADP-ribose polymerase expression, and by downregulating Bcl-2 expression. Moreover, casticin induced cycle arrest and apoptosis by upregulating p27 and downregulating cyclinD1/cyclin-dependent kinase4 and phosphorylated protein kinase B. In vivo, casticin inhibited tumor growth. Casticin induces G0/G1 arrest and apoptosis in gallbladder cancer, suggesting that casticin might represent a novel and effective agent against gallbladder cancer.

Background: Despite years of research, pancreatic ductal adenocarcinoma (PDAC) remains one of the deadliest cancers. A highly fibrotic, immunosuppressive microenvironment poses a significant barrier to effective immunotherapy. As key innate immune effectors of antitumor immunity, the functional status of natural killer (NK) cells within this environment remains poorly understood. This study aims to explore the role of stromal–immune interactions, particularly myofibroblastic cancer-associated fibroblasts (myCAFs) and NK cells, in shaping immune dysfunction in PDAC. Methods: We integrated spatial, single-cell and bulk transcriptomic analyses to characterize NK-cell heterogeneity and its cellular and molecular regulation in PDAC. CellChat-based communication analysis, spatial co-localization, and multiplex immunofluorescence were utilized to identify regulatory interactions and validate spatial associations. Results: A tumor-associated NK (TaNK) subset with impaired cytotoxicity was identified. myCAFs strongly interacted with TaNK cells through collagen signaling. Within ligand–receptor pairs, COL1A2–CD44 and COL1A1–LAIR1 served as dominant mediators linking stromal remodeling to impaired NK cell function. Spatial and histological validation confirmed their co-localization, while tumors co-enriched with myCAFs and TaNKs exhibited immune exclusion and poor response to immunotherapy. Conclusions: Our study demonstrates that collagen signaling from myCAFs impairs NK-cell cytotoxicity and promotes immune exclusion in PDAC. These findings advance the understanding of stromal–immune mechanisms underlying immunotherapy resistance and highlight that targeting the myCAF–NK axis may help improve the response to immunotherapy in PDAC.

Fail to reach the bilioenteric anastomosis is the main cause of treatment failure during single-balloon enteroscopy-assisted endoscopic retrograde cholangiopancreatography (SBE-assisted ERCP) in patients after bilioenteric Roux-en-Y anastomosis. We aim to evaluate factors influencing the endoscopic insertion failure. We retrospectively reviewed the clinical data of 231 cases undergoing SBE-assisted ERCP from January 2016 to December 2021. Treatment details and outcomes were studied to analyze the factors involved in endoscopic insertion failure. The enteroscopy success rate and procedural success rate were 88.3% and 84.4%. Incidence of postoperative adverse events was 3.9%. No serious adverse events occurred. Risk factors of endoscopic insertion failure include first ERCP attempt, side to side anastomosis at the Rou-Y anastomosis, the use of Endo-GIA anastomosis, three bowel lumens seen at the Rou-Y anastomosis under endoscopy, steep angle of the afferent loop at the Rou-Y anastomosis with a U-shape, length of the afferent loop ≥ 50 cm, and twisted afferent loop. Among which the multifactorial analysis suggested that the presence of three bowel lumens at the Rou-Y anastomosis and twisted afferent loop were independent risk factors for enteroscopy failure. For case with twisted afferent loop, the use of a transparent cap with X-ray-assisted guidance during insertion is an effective strategy to improve the success rate. SBE-assisted ERCP is safe and effective in patients after bilioenteric Roux-en-Y anastomosis. The severity of afferent loop twisting and Rou-Y anastomosis shape were risk factors for endoscopic insertion failure. Surgeons should take into account the feasibility of postoperative ERCP treatment at the time of operation.