Abstract Small interfering RNAs (siRNAs) have become a transformative class of nucleic acid therapeutics for clinical disease treatment, yet sequence-dependent off-target silencing continues to pose a major safety barrier that hinders their preclinical refinement and large-scale translational application. Existing bioinformatics tools only support partial off-target evaluation, either focusing on basic sequence optimization or simple seed-region scanning, and fail to deliver systematic, multi-dimensional and reproducible safety assessment for siRNA lead screening. To fill this gap, we developed TargetSureR, a lightweight, modular and CRAN-compatible open-source R package dedicated to full-process siRNA off-target risk profiling. This tool accommodates dual sequence-based and precomputed position-based inputs, integrates GTEx multi-tissue expression data and curated cancer, adverse-event and immune gene panels, and establishes a seven-dimensional scoring framework to stratify off-target risks into four hierarchical tiers. It further enables tissue-specific safety characterization and quantitative cross-species model selection, with an Ensembl API fallback mechanism ensuring high transcript annotation resolution. Built purely in R with no external shell dependencies, TargetSureR provides a standardized, robust and user-friendly workflow for systematic siRNA preclinical safety evaluation, and is freely available at https://github.com/nishuai/TargetSureR .

BackgroundCyclovirobuxine D (CVB-D), an alkaloid from Buxus microphylla, exhibits anticancer effects in various tumors, but its role in osteosarcoma remains unexplored. This study investigates its efficacy and mechanism in osteosarcoma.MethodsWe screened a drug library and evaluated CVB-D’s effects on osteosarcoma cell lines using CCK-8 and colony formation assays. Saos2 and K7M2 cells were selected for further analysis. Flow cytometry assessed apoptosis and cell cycle. RNA-seq identified downstream pathways, including NF-kappaB, and stemness markers (CD24, ALDH1A1). Stemness was examined via serum-free suspension culture, and NF-kappaB pathway activator Diprovocim was used in rescue experiments. A xenograft mouse model validated the findings.ResultsCVB-D suppressed proliferation, stemness, and induced apoptosis in osteosarcoma. These effects may be partially mediated through the p-NF-kappaB2/NF-kappaB2 axis and were reversible upon NF-kappaB pathway activation.ConclusionCVB-D inhibits osteosarcoma possibly via the non-canonical NF-kappaB pathway, suggesting a potential therapeutic strategy.

Background: The geriatric nutritional risk index (GNRI), a composite metric of serum albumin and body weight, has emerged as a prognostic tool in various cancers. However, its relevance in nasopharyngeal carcinoma (NPC) patients treated with volumetric modulated arc therapy (VMAT) remains unexplored. The aim of this study was to assess the effect of the GNRI in the prediction of the prognosis of nasopharyngeal carcinoma in the era of VMAT. Methods: This retrospective study analyzed 498 newly diagnosed, non-metastatic NPC patients treated with VMAT between 2010 and 2011. The GNRI was calculated using serum albumin and body weight ratios, with receiver operating characteristic (ROC) curve analysis determining its optimal prognostic cutoff. Patients were stratified into training (70%) and validation (30%) cohorts. Cox regression identified independent prognostic factors, which were integrated into a nomogram predicting 3- and 5-year overall survival (OS). Model performance was assessed via the concordance index (C-index), calibration curves, and decision curve analysis (DCA). Results: In the study, 348 patients were included in the training cohort and 150 patients were included in the validation cohort according to a ratio of 7:3. The median follow-up was 68 months, with 5-year OS rates of 79.3%. A GNRI > 102 independently predicted improved survival (HR = 0.64; p = 0.044), alongside tumor volume, age, and N-stage. The nomogram demonstrated strong discrimination (C-index: 0.757–0.762 for training; 0.737–0.744 for validation) and calibration, aligning closely with observed survival. DCA confirmed superior clinical utility over default strategies. NPC patients treated with VMAT with a high GNRI, female sex, and a lower N-stage exhibited significantly better OS (p < 0.05). Conclusions: The GNRI is a robust prognostic marker for NPC patients receiving VMAT, reflecting the interplay of nutrition, inflammation, and treatment response. The validated nomogram provides a practical tool for individualized risk stratification, enhancing clinical decision-making in the era of precision radiotherapy.

BACKGROUND:The FLASH effect is dose-dependent, and fractional dose optimization may enhance it, improving normal tissue sparing. PURPOSE:This study investigates the performance of fractional dose optimization in enhancing normal tissue sparing for Bragg peak FLASH radiotherapy (FLASH-RT). METHODS:15 lung cancer patients, including eight with peripherally located tumors and seven with centrally located tumors, were retrospectively analyzed. A uniform fractionation prescription of 50 Gy in five fractions was utilized, corresponding to a biological equivalent dose (BED) of 100 Gy, calculated using an α/β value of 10 Gy. For each patient, uniform (UFD) and nonuniform fractional dose (non-UFD) plans were designed. In UFD FLASH plans, five multi-energy Bragg peak beams were optimized using single-field optimization, each delivering 10 Gy to the target. In non-UFD FLASH plans, fractional doses were optimized to enhance sparing effects while ensuring the target received a BED comparable to UFD plans. A dose-dependent FLASH enhancement ratio (FER) was integrated with the BED to form the FER-BED metric to compare the UFD and non-UFD plans. An α/β value of 3 Gy was applied for normal tissues in the calculations. RESULTS:Bragg peak FLASH plans showed high dose conformality for both peripheral and central tumors, with all plans achieving a conformality index (the ratio of the volume receiving the prescribed dose to the CTV volume) below 1.2. In non-UFD plans, fractional doses ranged from 5.0 Gy to 20.0 Gy. Compared to UFD plans, non-UFD plans achieved similar BED coverage (BED98%: 96.6 Gy vs. 97.1 Gy, p = 0.256), while offering improved organ-at-risk sparing. Specifically, the FER-BED15cc for the heart reduced by 10.5% (9.4 Gy vs. 10.5 Gy, p = 0.017) and the V6.7GyFER-BED for the ipsilateral lung decreased by 4.3% (29 .1% vs. 30.4%, p = 0.008). No significant difference was observed in FER-BED0.25cc of spinal cord (UFD: 7.1 Gy, non-UFD: 6.9 Gy, p = 0.626) and FER-BED5cc in esophagus (UFD: 0.4 Gy, non-UFD: 0.4 Gy, p = 0.831). CONCLUSIONS:Bragg peak FLASH-RT achieved high dose conformality for both peripheral and central tumors. Fractional dose optimization, using a single beam per fraction delivery mode, enhanced normal tissue sparing by leveraging both fractionation and FLASH effects.

This paper aims to propose a noninvasive radiotherapy patient positioning system based on structured light surface imaging, and evaluate its clinical feasibility. First, structured light sensors were used to obtain the panoramic point clouds during radiotherapy positioning in real time. The fusion of different point clouds and coordinate transformation were realized based on optical calibration and pose estimation, and the body surface was segmented referring to the preset region of interest (ROI). Then, the global-local registration of cross-source point cloud was achieved based on algorithms such as random sample consensus (RANSAC) and iterative closest point (ICP), to calculate 6 degrees of freedom (DoF) positioning deviation and provide guidance for the correction of couch shifts. The evaluation of the system was carried out based on a rigid adult phantom and volunteers' body, which included positioning error, correlation analysis, and receiver operating characteristic (ROC) analysis. Using Cone Beam CT (CBCT) as the gold standard, the maximum translation and rotation errors of this system were (1.5 ± 0.9) mm along Vrt direction (chest) and (0.7 ± 0.3) ° along Pitch direction (head and neck). The Pearson correlation coefficient between results of system outputs and CBCT verification distributed in an interval of [0.80, 0.84]. Results of ROC analysis showed that the translational and rotational AUC values were 0.82 and 0.85, respectively. In the 4D freedom accuracy test on the human body of volunteers, the maximum translation and rotation errors were (2.6 ± 1.1) mm (Vrt direction, chest and abdomen) and (0.8 ± 0.4)° (Rtn direction, chest and abdomen) respectively. In summary, the positioning system based on structured light body surface imaging proposed in this article can ensure positioning accuracy without surface markers and additional doses, and is feasible for clinical application.

OBJECTIVE:Pulmonary fibrosis(PF) is an abnormal wound-healing response resulting from recurrent alveolar injury, characterized by persistent inflammation and excessive collagen deposition. Given the limited clinical treatment options, novel therapeutic strategies are urgently needed. Urolithin A (UA), a secondary metabolite produced by intestinal microbiota from natural polyphenols, has attracted attention due to its anti-inflammatory, antioxidant, and anti-aging properties. However, the therapeutic efficacy and mechanisms of UA in PF remain unclear. This study aimed to investigate the protective effects and underlying molecular mechanisms of UA in PF. METHODS:This study integrated network pharmacology analysis, molecular docking, and in vitro/in vivo experiments to elucidate the anti-fibrotic mechanisms of UA. Firstly, a mouse model of PF was established via intratracheal instillation of bleomycin. Mice in the UA treatment group received daily oral administration of UA (20 mg/kg/day) starting on day 10 post-modeling and continuing until day 21, at which point lung tissues were collected. Histopathological alterations and collagen deposition in the lungs were assessed using Masson's trichrome staining and hydroxyproline content analysis. Furthermore, network pharmacology was employed to predict the potential molecular targets and pathways of UA, which were further validated through molecular docking and in vitro fibroblast experiments to verify the underlying mechanisms. RESULTS:UA treatment significantly alleviated PF in mice, evidenced by reduced collagen deposition, diminished structural damage, and notably decreased excessive extracellular matrix accumulation. Network pharmacology analysis and molecular docking indicated that the PI3K/AKT/mTOR signaling pathway is the primary pharmacological target of UA's anti-fibrotic effect. Further in vitro experiments demonstrated that UA significantly suppressed fibroblast activation by inhibiting AKT1 phosphorylation. Moreover, the inhibitory effects of UA on fibroblasts were reversed upon reactivation of the AKT pathway by the AKT agonist SC79, further confirming the crucial role of the AKT signaling pathway in UA's anti-fibrotic mechanism. CONCLUSION:UA exerts therapeutic effects on PF by targeting the PI3K/AKT/mTOR pathway, particularly through the inhibition of AKT1 phosphorylation. These findings indicate that UA has potential as a therapeutic candidate for PF and provide a novel perspective for utilizing gut microbiota metabolites in the treatment of fibrotic diseases. Further studies are needed to elucidate the precise mechanisms of UA.

Di-(2-ethylhexyl) phthalate (DEHP), a ubiquitous plasticizer in consumer and medical products, readily leaches into the environment and is metabolized into its bioactive derivative mono-(2-ethylhexyl) phthalate (MEHP). Emerging evidence links DEHP/MEHP exposure to tumor progression and drug resistance in multiple cancers. While environmental factors are hypothesized to contribute to the pathogenesis of multiple myeloma (MM)-a currently incurable plasma cell malignancy-the specific role of DEHP and MEHP in MM remains unexplored. This study aimed to investigate the effects of prolonged DEHP exposure on MM progression and drug resistance. Comparative analysis of MM cells with and without DEHP exposure demonstrated that DEHP promotes tumorigenesis and induces resistance to bortezomib, a frontline MM therapy. Mechanistically, DEHP activates the Wnt/β-catenin and NF-κB signaling pathways, and MEHP recapitulates these pro-tumorigenic effects. Our findings not only identify DEHP/MEHP as potential environmental risk factors for MM but also implicate Wnt/β-catenin and NF-κB as actionable targets for therapeutic intervention. These results underscore the need to reevaluate DEHP exposure risks in clinical settings and public health policies.

Exosomes can be used to mediate the delivery of nucleic acids such as microRNA-125b-5p (miR-125b-5p), a tumor-suppressor in certain types of cancer, into tumor cells. The present study investigated the use of bone mesenchymal stem cells-derived exosome (BMSCs-Exo) delivery of miR-125b-5p in ovarian cancer (OC). BMSCs were transfected with miR-125b-5p mimic, from which exosomes termed Exo-miR-125b-5p mimic were extracted. The expression levels of miR-125b-5p in OC tissue samples, BMSCs, exosomes and SKOV3 cells were quantified using reverse transcription-quantitative PCR. The influence of Exo-miR-125b-5p mimic on the biological functions of OC was evaluated through cell proliferation, invasion, migration and apoptosis assays. The targeting relationship between miR-125b-5p and DEAD-box helicase 5 (DDX5) was verified, and the expression levels of DDX5 in OC samples and SKOV3 cells were quantified using western blotting. miR-125b-5p was downregulated in tumor tissue samples from patients with OC. BMSCs-Exo reduced the malignant properties of SKOV3 cells in vitro, and these effects were be advanced by miR-125b-5p upregulation. miR-125b-5p targeted and inhibited DDX5 expression. DDX5 overexpression inhibited Exo-miR-125b-5p-induced suppression of OC development. Overall, this study highlights that BMSCs-Exo-encapsulated miR-125b-5p inhibited OC progression via DDX5 downregulation, providing insight into the molecular mechanisms underlying OC.

Abstract Purpose This research designeded to: 1. Analyze the efficacy and safety of Palbociclib treatment in HR-positive and HER2-negative (HR + /HER2-) metastatic breast cancer(MBC) patients. 2. Establish and validate a nomogram model for predicting the progression-free survival (PFS) rates of 6 months, 12 months, and 18 months in HR + /HER2- MBC patients after receiving Palbociclib plus endocrine therapy (ET). Patients and methods 1. This research retrospectively analyzed the efficacy and safety of Palbociclib combined with ET in 214 patients with HR + /HER2- MBC. 2. A nomogram was designed and constructed with the retrospective clinical data of 214 patients with HR + /HER2- MBC who received Palbociclib plus ET at Zhejiang Cancer Hospital in China from August 2018 to August 2022. Among these patients, 161 were randomly assigned to the training cohort, while 53 to the validation cohort. The predictive accuracy of the nomogram was assessed through the analysis the area under the receiver operating characteristic(ROC) curve, calibration curve, and decision curve analysis(DCA). Results 1. Median PFS was 7.17 months (95% CI: 7.61—10.05 months), with an objective response rate (ORR) of 2.80% and a disease control rate (DCR) of 34.58%. The most prevalent grade 3–4 adverse event was neutropenia (38.79%). 2. Multiple variable analysis of the training set revealed that age < 60 years old, PR < 20%, Ki-67 ≥ 20%, luminal B molecular subtype, primary resistance to ET, receipt of late-stage chemotherapy, and presence of liver metastasis or ≥ 2 visceral metastases were independent prognostic factors associated with poor PFS (P < 0.05). Then, the predictive model underwent development and validation utilizing the aforementioned parameters. On the one hand, the area under the ROC curve (AUC) values of the training set at 6 months, 12 months, and 18 months were 0.771, 0.783, and 0.790, respectively, indicating a strong predictive ability of the developed model. On the other hand, the AUC of the validation set at 6 months, 12 months, and 18 months were 0.720, 0.766, and 0.754, respectively, suggesting the favorable discriminatory ability of the model. The calibration curves also exhibited a good fit with the ideal curves, and the DCA demonstrated the clinical applicability of the model. The nomogram's different scores could distinguish PFS. Conclusion This retrospective study demonstrates the efficacy of Palbociclib in Chinese breast cancer patients. Moreover, the clinical parameters showed a significant association with the prognosis of HR + /HER2- MBC, and the prognostic models constructed based on these variables also displayed robust predictive power, which could offer more intuitive and convenient references for clinical doctors to formulate follow-up treatment plans.

To investigate the differences between lung adenocarcinoma with the pleural invasion that has EGFR (epidermal growth factor receptor) 19-del or 21L858R mutations in terms of clinical characteristics and outcomes. EGFR mutation-positive patients with pleural metastasis of lung adenocarcinoma diagnosed in the Department of Respiratory Medicine of Yuhuangding Hospital of Yantai City, Shandong Province, from January 2014 to January 2022 were selected. The clinical data of the patients were collected to retrospectively analyze whether the clinical characteristics and prognosis of patients with 19-del or 21L858R mutation subtype were different and analyze the impact of clinical characteristics on the prognosis of patients. The difference in clinical characteristics between the 2 groups was analyzed by SPSS, P < .05. There was statistical significance. Univariate and multivariate regression analysis was performed with R soft. To establish a 2-year overall survival predictive model for patients with EGFR gene 19-del and 21L858R mutations in patients with pleural invasion of lung adenomas and to provide predictive model maps. Receiver operating characteristic curve, calibration curve, and decision curve analysis were used to evaluate the value of the prediction model in this study. Of the 74 patients included, the 19-del mutation group had a higher incidence of pleural thickening (P = .023) and a lower Ki-67 level (P = .035). There was no difference in 2-year overall survival and progression-free survival between the 2 mutations. There were differences in pleural thickening and Ki-67 index between the 2 groups, but no differences in disease outcome between the 2 groups. The nomogram model established based on gender, treatment regimen, CEA, lymph node metastasis, and pleural changes is accurate and feasible.

In 2021, breast cancer accounted for a substantial proportion of cancer cases and represented the second leading cause of cancer deaths among women worldwide. Although tumor cells originate from normal cells in the human body, they possess distinct biological characteristics resulting from changes in gene structure and function of cancer cells in contrast with normal cells. These distinguishing features, known as hallmarks of cancer cells, differ from those of normal cells. The hallmarks primarily include high metabolic activity, mitochondrial dysfunction, and resistance to cell death. Current evidence suggests that the fundamental hallmarks of tumor cells affect the tissue structure, function, and metabolism of tumor cells and their internal and external environment. Therefore, these fundamental hallmarks of tumor cells enable tumor cells to proliferate, invade and avoid apoptosis. Modifying these hallmarks of tumor cells represents a new and potentially promising approach to tumor treatment. The key to breast cancer treatment lies in identifying the optimal therapeutic agent with minimal toxicity to normal cells, considering the specific types of tumor cells in patients. Some herbal medicines contain active ingredients which can precisely achieve this purpose. In this review, we introduce Ginsenoside's mechanism and research significance in achieving the therapeutic effect of breast cancer by changing the functional hallmarks of tumor cells, providing a new perspective for the potential application of Ginsenoside as a therapeutic drug for breast cancer.

The PHLPP1/AKT/HIF-1A/VEGF-C axis does not exist in EBV-associated GC

Recent studies have demonstrated that the combination of Cyclin-Dependent Kinase 4/6 Inhibitor (CDK4/6i) and endocrine therapy (ET) is more effective than ET alone and significantly improves progression-free survival (PFS) and overall survival (OS) in patients with hormone receptor-positive (HR+)/human epidermal growth factor receptor-2 negative (HER2-) breast cancer (BC). Palbociclib is the first CDK4/6i approved for use, and its clinical advantages have been shown. However, 30% of patients will continue to develop secondary drug resistance. Therefore, exploring the parameters that can predict the efficacy of Palbociclib and developing a clinical prediction model is essential for evaluating the prognosis of patients.

Background Previous researches have demonstrated that adaptive replanning during intensity-modulated radiation therapy (IMRT) could enhance the prognosis of patients with nasopharyngeal carcinoma (NPC). However, the delineation of replanning target volumes remains unclear. This study aimed to evaluate the feasibility of reducing target volumes through adaptive replanning during IMRT by analyzing long-term survival outcomes and failure patterns of locoregional recurrence in NPC. Methods This study enrolled consecutive NPC patients who received IMRT at our hospital between August 2011 and April 2018. Patients with initially diagnosed, histologically verified, non-metastatic nasopharyngeal cancer were eligible for participation in this study. The location and extent of locoregional recurrences were transferred to pretreatment planning computed tomography for dosimetry analysis. Results Among 274 patients, 100 (36.5%) received IMRT without replanning and 174 (63.5%) received IMRT with replanning. Five-year rates of locoregional recurrence-free survival (LRFS) were 90.1% (95%CI, 84.8% to 95.4%) and 80.8% (95%CI, 72.0% to 89.6%) for patients with and without replanning, P = 0.045. There were 17 locoregional recurrences in 15 patients among patients with replanning, of which 1 (5.9%) was out-field and 16 (94.1%) were in-field. Among patients without replanning, 19 patients developed locoregional recurrences, of which 1 (5.3%) was out-field, 2 (10.5%) were marginal, and 16 (84.2%) were in-field. Conclusions In-field failure inside the high dose area was the most common locoregional recurrent pattern for non-metastatic NPC. Adapting the target volumes and modifying the radiation dose prescribed to the area of tumor reduction during IMRT was feasible and would not cause additional recurrence in the shrunken area.

EBV is not correlated with lymphangiogenesis in GC

Identification of EBV infection and genome clearance efficiency in the EBV-negative and positive-NPC cell lines

Radiotherapy (RT) is one of the most widely used treatment strategies in cancer treatment in non-small cell lung cancer (NSCLC), but radioresistance remains a major clinical challenge. Ferroptosis is a recently identified form of non-apoptosis regulated cell death (RCD) driven by iron-dependent iron-dependent lipid peroxidation, and recent studies showed ferroptosis plays an important role in RT-induced cell death. However, the dysregulation of ferroptosis in radioresistance is poorly understood.

Abstract Lymphatic metastasis is a common clinical symptom in nasopharyngeal carcinoma (NPC), the most common Epstein-Barr virus (EBV)-associated head and neck malignancy. However, the effect of EBV on NPC lymph node (LN) metastasis is still unclear. In this study, we demonstrated that EBV infection is strongly associated with advanced clinical N stage and lymphangiogenesis of NPC. We found that NPC cells infected with EBV promote LN metastasis by inducing cancer-associated lymphangiogenesis, whereas these changes were abolished upon clearance of EBV genomes. Mechanistically, EBV-induced VEGF-C contributed to lymphangiogenesis and LN metastasis, and PHLPP1, a target of miR-BART15, partially contributed to AKT/HIF1a hyperactivity and subsequent VEGF-C transcriptional activation. In addition, administration of anti-VEGF-C antibody or HIF1α inhibitors attenuated the lymphangiogenesis and LN metastasis induced by EBV. Finally, we verified the clinical significance of this prometastatic EBV/VEGF-C axis by determining the expression of PHLPP1, AKT, HIF1a, and VEGF-C in NPC specimens with and without EBV. These results uncover a reasonable mechanism for the EBV-modulated LN metastasis microenvironment in NPC, indicating that EBV is a potential therapeutic target for NPC with lymphatic metastasis. Implications: This research demonstrates that EBV induces lymphangiogenesis in NPC by regulating PHLPP1/p-AKT/HIF1a/VEGF-C, providing a new therapeutic target for NPC with lymphatic metastasis.