Nuclearfactor erythroidderived 2-like 2(NRF2) is a major regulator of the body's antioxidant defense system and a key protein in the process of ferroptosis, which is upregulated in a variety of malignancies. This study aimed to investigate the role of NRF2 in the pathogenesis and progression of Esophageal squamous cell carcinoma (ESCC) from the in vivo and clinical levels. The expression levels of NRF2, Glutamate-Cysteine Ligase Modifier Subunit (GCLM) and Glutathione Peroxidase 4 (GPX4) in ESCC and adjacent normal tissues were detected by immunohistochemistry in 61 tissue biopsies collected from patients diagnosed with ESCC. The xenograft model was used to detect the growth of nude mouse tumors and the changes of ferroptosis-related indexes in different experimental groups. Co-immunoprecipitation was used to demonstrate the downstream interacting proteins of NRF2. The results showed that the expression of NRF2, GCLM and GPX4 was increased in ESCC compared with adjacent non-tumor tissues, and the high expression of NRF2, GCLM and GPX4 was significantly associated with poor prognosis. NRF2 overexpression promotes changes in tumor growth and ferroptosis-related markers in xenograft models. In addition, NRF2 overexpression was associated with upregulation of GCLM and GPX4. CO-IP demonstrated that GCLM is a downstream protein of NRF2. Bio-informatics analysis showed that GCLM was differentially expressed in a variety of tumors, and was significantly correlated with the prognosis of patients, as well as the infiltration of a variety of immune cells. Finally, GCLM promotes tumor growth and radiotherapy resistance to ESCC in vivo, and can therefore be used as a molecular target for tumor therapy.

Immune checkpoint inhibitors (ICIs), either as monotherapy (ICI-Mono) or combined with chemotherapy (ICI-Chemo), improves survival in advanced non-small cell lung cancer (NSCLC). However, prospective guidance for choosing between these options remains limited, and single-feature biomarkers like PD-L1 prove inadequate. We develop a machine learning model using clinicogenomic data from four cohorts (MD Anderson n = 750; Mayo Clinic n = 80; Dana-Farber n = 1077; Stand Up To Cancer n = 393) to predict individual benefit from adding chemotherapy. Benefit scores are calculated using five distinct functions derived from 28 genomic and 6 clinical features. Our integrated model, A-STEP (Attention-based Scoring for Treatment Effect Prediction), estimates heterogeneous treatment effects and achieves the largest reduction in 3-month progression risk, improving weighted risk reduction by 13-23% over stand-alone models. A-STEP recommends treatment changes for over 50% of patients, most often favoring ICI-Chemo. In simulation on external cohort, patients treated in accordance with A-STEP recommendations show improved 2-year progression-free survival (HR = 0.60 for ICI-Mono treatment arm; HR = 0.58 for ICI-Chemo treatment arm). Predictive features include FBXW7, APC, and PD-L1. In this study, we demonstrate how machine learning can fill critical gaps in immunotherapy selection for NSCLC, by modeling treatment heterogeneity with real-world clinicogenomic data, driving precision medicine beyond conventional biomarker boundaries.

It was the first time that a spectrofluorimetric approach for the simultaneous analysis of methotrexate (MET) and 6-mercaptopurine (6-MP) had been achieved. This study aims to propose an accurate and sensitive second derivative synchronous spectrofluorimetric approach for the measurement of MET and 6-MP in different matrices simultaneously without the need for prior separation. The proposed approach measured the synchronous fluorescence intensity of pharmaceuticals under research at a constant wavelength difference (Δλ) = 130 nm. For the quantitative analysis of 6-MP and MET, the second derivative's peak amplitudes were observed at 380 and 400 nm, respectively. The process was fully validated and showed exceptional linearity for either 6-MP or MET in the concentration range of 10-100 ng mL^-1. The new method was used to quantitatively analyze both drugs in a synthetically formulated mixture, dosage forms, and spiked human plasma. A statistical comparison of the results with other published analytical techniques revealed no significant difference. The validation of the procedure was successfully completed in compliance with ICH guidelines. To assess the analytical methodology's greenness, Eco-Scale and GAPI greenness tools were employed.

Locally advanced colorectal cancer patients are characterized by poor prognosis and high recurrence rates, with peritoneal metastasis rates as high as 20%-30%. Despite curative resection and chemotherapy being the main treatment methods, challenges remain in preventing peritoneal metastasis. Neoadjuvant therapy and immunotherapy are hot topics of research, and hyperthermic intraperitoneal chemotherapy (HIPEC) is one of the new approaches for preventing peritoneal metastasis, yet its value and safety are still controversial. HIPEC can directly target free tumor cells in the abdominal cavity through hyperthermic effects and high concentrations of chemotherapeutic drugs, but its prophylactic use requires further exploration regarding effectiveness and risks. Early intervention and identification of high-risk factors are crucial for improving therapeutic outcomes, and tests such as circulating tumor DNA and free peritoneal cell DNA provide new avenues for early screening. The value of prophylactic HIPEC varies across different studies, and its complications and risks should not be overlooked. The selection of chemotherapy drugs, dosage, and personalized treatment plans are key factors affecting therapeutic efficacy. Other prevention strategies, such as pressurized intraperitoneal aerosol chemotherapy and neoadjuvant chemotherapy, are also being explored. In summary, prophylactic HIPEC shows some potential in controlling peritoneal metastasis, but its application requires individualized assessment and optimization.

Immunotherapy based on immune checkpoint inhibitor (ICI) has shown remarkable efficacy in the treatment of microsatellite instability (MSI)-high CRC. However, the monotherapy of ICI in microsatellite stable (MSS) CRC has not been satisfactory. Some patients with MSS CRC can benefit from various combination immunotherapy regimens. Identifying appropriate biomarkers to select MSS-type CRC patients who will benefit from ICI treatment prior to therapy initiation and dynamically monitoring treatment efficacy during the therapeutic course have become crucial components of precision medicine in clinical practice. This article reviews the current research status of traditional biomarkers such as tumor mutation burden (TMB) and PD-L1 expression. It also explores the latest research progress and clinical translation potential of emerging biomarkers, including POLE/POLD1 mutations, immune score, circulating tumor DNA, and gut microbiome. Furthermore, it addresses the challenges in the clinical application of biomarkers, such as the controversy over TMB cutoff values and the heterogeneity of PD-L1 expression. Finally, it outlines future research directions with the aim of providing a basis for clinical decision-making in immunotherapy and facilitating the realization of precision medicine.

Total mesorectal excision (total mesorectal excision, TME) has currently become the standard procedure for the treatment of rectal cancer. However, whether to preserve the left colic artery (LCA) during TME surgery remains controversial. The debates mainly focus on whether preserving the LCA can achieve complete dissection of No. 253 lymph nodes and its impacts on patients' defecation, urination, and sexual functions, and prognosis. This article systematically reviews the recent research progress necessity surgical techniques for LCA preservation by combining the anatomical characteristics of the inferior mesenteric artery (IMA) and LCA with clinical experience.

Total pelvic exenteration (TPE) is a critical surgical procedure for treating locally advanced rectal cancer (LARC) at stage T4b and locally recurrent rectal cancer (LRRC), where the resectability of pelvic tumors depends on precise preoperative imaging evaluation. Laparotomy primarily aims to exclude abdominopelvic peritoneal metastases. Preoperative assessment involves contrast-enhanced chest/abdominal CT, contrast-enhanced liver MRI (as a supplement when CT findings are unclear), contrast-enhanced pelvic MRI (the preferred modality for evaluating soft tissue planes, organ involvement, and resection scope), and PET-CT (useful for systemic metastasis detection and differentiating scar/fibrosis from tumor). Key focuses include identifying invasion of pelvic wall structures (vascular, neural, muscular planes in lateral, posterior, and floor regions) and the "high-risk zone for major hemorrhage" at the confluence of internal iliac veins. Multidisciplinary team discussions involving radiology, surgery, oncology, and other specialties are essential. These discussions emphasize "en bloc resection" principles, using imaging to define resection planes layer-by-layer to assess R0 resection feasibility, reconstructive strategies, and neoadjuvant therapy. The "Changzheng Surgical Classification" proposed by our center categorizes PE into intra-pelvic exenteration (resecting ≥50% of tissues from ≥2 systems within the bony pelvis) and combined pelvic wall exenteration (involving ≥50% tissues from ≥1 pelvic system plus ≥1 of the 5 pelvic wall regions or ≥2 pelvic wall regions). Preoperative planning based on detailed pelvic anatomical zoning ensures standardized resection and reconstruction, promoting procedural consistency and improving R0 resection rates.

Tianhe procedure^® is a functional sphincter-preserving surgical approach developed for rectal cancer patients following radiotherapy. This technique involves extended proximal resection of the colon beyond the pelvic cavity, followed by anastomosis of the non-irradiated proximal colon to the distal rectum or anal canal. This strategy aims to reduce the incidence of anastomotic complications and postoperative bowel dysfunction. However, there is currently a lack of standardized practice guideline for implementing Tianhe procedure^® in China. Therefore, the Chinese Radiation Intestinal Injury Research Group, the Colorectal Surgery Group of Surgery Branch of the Chinese Medical Association, the Anorectal Branch of Chinese Medical Doctor Association, the Colorectal Cancer Committee of the Chinese Medical Doctor Association, and the Colorectal Cancer Committee of China Anti-cancer Association, and the Gastrointestinal Surgical Branch of Guangdong Medical Doctor Association, have jointly convened a panel of national experts to discuss and establish this standardized surgical procedure. This standard, based on the latest evidence from literature, research advancements, and expert experience, focuses on key aspects of the Tianhe procedure^®, including its precise definition, indications, critical procedural steps, postoperative complications, and functional rehabilitation strategies. It aims to promote standardized implementation and broader clinical adoption of this innovative surgical technique.

This study aims to investigate the therapeutic efficacy and molecular mechanism of cinobufagin in non-small cell lung cancer (NSCLC) via pyroptosis induction. Bronchial epithelial cells and NSCLC cell lines were treated with gradient concentrations of cinobufagin. Cell viability was evaluated using Cell Counting Kit-8 (CCK-8) assay. RNA-sequencing was performed to identify differentially expressed genes. Lactate dehydrogenase (LDH) release was measured via cytotoxicity detection kit. Pyroptotic morphological changes were observed by transmission electron microscopy. Western blotting analysed expression levels of pyroptosis-related proteins. In vivo efficacy was validated in nude mouse xenograft models. Immunohistochemistry evaluated tumour pyroptosis markers, whilst flow cytometry analysed tumour-infiltrating CD8⁺ T cells and natural killer (NK) cells. Cinobufagin demonstrated selective cytotoxicity against NSCLC cells with minimal toxicity to normal bronchial epithelium. RNA-seq analysis revealed significant enrichment of pyroptosis-related pathways. Functional experiments confirmed cinobufagin-induced LDH release, characteristic pyroptotic morphological changes and upregulation of cleaved caspase-3 and Gasdermin E (GSDME)-NT in NSCLC cells. In xenograft models, cinobufagin treatment reduced tumour volume compared to controls. Mechanistically, this was associated with enhanced caspase-3 activation and GSDME-NT accumulation in tumour tissues. Notably, cinobufagin treatment significantly increased NK cell infiltration and activity. Cinobufagin exerts antitumor effects in NSCLC through caspase-3/GSDME-mediated pyroptosis induction, accompanied by immune microenvironment modulation. These findings provide preclinical evidence for cinobufagin as a potential therapeutic agent targeting pyroptosis in NSCLC.

Patients with head and neck squamous cell carcinoma (HNSCC), particularly the human papillomavirus negative (HPV^-) subset, have a dismal prognosis. Furthermore, patients with Fanconi anemia (FA) have a genetic predisposition with a 500-fold to 700-fold higher incidence of HNSCC. Thus, novel and more efficacious therapies are needed. As current immunotherapies often fail due to suppressive elements in the tumor microenvironment (TME), we developed a trispecific killer engager (TriKE) to direct multiple signals to natural killer (NK) cells to overcome the hypoxic TME. This TriKE is comprised of a camelid nanobody that binds to CD16 on NK cells, an interleukin (IL)-15 moiety, and another novel camelid nanobody that binds to the B7-H3 antigen, which is highly and specifically expressed on the tumor cell surface.

The B7H3 TriKE was generated using a mammalian expression system. Its functionality was evaluated using flow cytometry-based NK cell degranulation, cytokine production, proliferation and live cell imaging cytotoxicity assays. Models of acute and prolonged hypoxia (1% oxygen) were carried out to assess tumor killing. Tumor progression, NK cell persistence, and survival differences between IL-15-treated and TriKE-treated mice were studied using NOD-scidIL2Rg^null (NSG) mice engrafted with human HNSCC.

High B7-H3 expression was found in HPV^- HNSCC cell lines, even when the FA gene was knocked out, and The Cancer Genome Atlas patient data showed that high B7-H3 expression predicted poor survival in patients with HPV^- HNSCC. Similar to the NK cell activity seen with healthy donors, the B7H3 TriKE enhanced activation, expansion and cytotoxicity of NK cells from patients with HPV^- HNSCC, a target population for this therapeutic. Additionally, the B7H3 TriKE improved NK cell cytotoxicity in a three-dimensional spheroid model of HNSCC. In both acute and prolonged hypoxia (1% oxygen), the B7H3 TriKE mediated enhanced tumor killing, mitigating impairment of NK cell cytotoxicity in hypoxia. In vivo, the B7H3 TriKE-treated mice demonstrated substantial antitumor activity and prolonged survival.

The B7H3 TriKE is a novel immunotherapeutic approach that can overcome hypoxic suppression of NK cells in the HNSCC TME. These highly translational studies present an innovative therapy for patients with HNSCC and will be developed further for clinical application.