Hepatocellular carcinoma (HCC) is a highly lethal malignancy with high invasiveness and metastasis. Despite progress in its treatment, the high mortality rate persists due to poor prognosis. In this study, we aimed to develop a prognostic model for HCC using platelet-related genes. We downloaded and preprocessed data from the Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) databases. Through differential gene expression analysis, univariate Cox regression, Least absolute shrinkage and selection operator (LASSO) machine learning, and multivariate Cox regression, we constructed a prognostic model consisting of six genes (KIF18A, HRG, TUBA4B, MAFF, SELP, and IGF1). The survival analysis and ROC curve evaluation conducted on both the training and validation sets showed excellent predictive performance of the model. In addition, the model was also associated with the malignancy and risk of metastasis of tumors. Additionally, analysis of half maximal inhibitory concentration (IC50) values between high- and low-risk groups for various drugs showed significant differences, suggesting potential differences in predicted drug sensitivity between risk groups. We identified potential stem-like cell subpopulations in HCC cells, which are mostly in the late stage of malignant cell differentiation. In conclusion, we successfully constructed a novel platelet-related prognostic model for HCC.

Pancreatic cancer (PC) is a highly aggressive malignancy, and zinc ion homeostasis is closely linked to its metastatic potential, though the underlying regulatory mechanisms remain unclear. To investigate this, single-cell RNA sequencing data from primary and metastatic PC were downloaded from the GEO database to analyze differences in zinc ion levels and identify key zinc-regulating proteins. A PC mouse model was further established, and experiments including immunofluorescence, immunohistochemistry, and zinc ion level detection were conducted to comprehensively evaluate the impact of zinc ions and their regulatory proteins on PC metastasis and the associated mechanisms. PC cell models with SLC39A1 knockdown or overexpression were also constructed; Western blot was used to measure protein expression levels, while cell scratch assay, Transwell invasion assay, and 3D cell sphere formation assay were employed to assess cell migration, invasion, and sphere-forming capabilities, respectively. Single-cell sequencing analysis demonstrated that zinc ions and SLC39A1 were significantly enriched in PC metastases, with higher enrichment levels in epithelial cells compared to other cell types. Overexpression of SLC39A1 increased intracellular zinc levels and enhanced the migration and invasion of PC cells, whereas application of zinc chelators or Src/FAK inhibitors effectively suppressed cancer cell migration and invasion induced by elevated zinc levels. Animal model experiments further showed that knockdown of SLC39A1 in PC cells inhibited their liver metastasis capability. Collectively, these findings indicated that SLC39A1 was highly expressed in PC metastases, where its elevated expression increased intracellular zinc levels and activated the Src/FAK signaling pathway, thereby promoting the migration and invasion of PC cells.

Aim: Colorectal cancer (CRC) ranks among the most prevalent malignancies across the globe, with treatment resistance often closely linked to the complexity of the tumor microenvironment (TME). This study aims to establish a gene signature associated with epithelial-mesenchymal transition (EMT) that integrates TME dynamics, prognosis prediction, and drug resistance assessment in CRC. Methods: We employed the Cancer Genome Atlas (TCGA) resource and bulk RNA-sequencing profiles linked to EMT to identify common differentially expressed genes (DEGs). An eight-gene signature was constructed using multivariable Cox regression analysis. The correlations of risk groups (scores) with overall survival, biological characteristics, and drug sensitivity were analyzed in CRC patients. Results: Patients in the high-risk group exhibited significantly worse clinical outcomes than those in the low-risk group. Moreover, a lower risk score was significantly correlated with increased responsiveness to both immune checkpoint inhibitors and 5-fluorouracil in CRC patients. Additionally, experiments in vitro and in vivo confirmed that FABP4 functions as an oncogene in CRC by facilitating cell growth, migration, and chemotherapy resistance. Conclusion: The EMT-associated gene signature holds significant value for predicting clinical prognosis, immunotherapy responsiveness, and chemotherapy sensitivity in CRC.

Auxin herbicides exert herbicidal effects by over-inducing auxin response in susceptible plants. This study investigated the molecular interactions between TIR1-IAA7 and distinct chemical classes of auxin herbicides. MCPA (2-methyl-4-chlorophenoxy acetic acid), chloramben (3-Amino-2,5-dichlorobenzoic acid), quinclorac (3,7-dichloro-8-quinolinecarboxylic acid), and aminocyclopyrachlor (6-amino-5-chloro-2-cyclopropylpyrimidine-4-carboxylic acid) exhibited relatively lower docking energies among all the 8 candidate auxin herbicides in the molecular docking simulation and were selected as representatives. In vitro 3H-IAA-binding and Surface Plasmon Resonance experiments indicated MCPA and chloramben showed stronger affinities for TIR1-IAA7, further corroborated by mutant resistance assays. Despite their weak affinity for TIR1-IAA7, quinclorac and aminocyclopyrachlor are likely to interact with an alternative AFB protein in combination with IAA7, thereby disrupting auxin signaling and down-regulate photosynthetic gene expression. These findings demonstrate that different herbicide classes exhibit distinct co-receptor preferences. Such target specificities provide a structural basis for the rational design of novel auxinic herbicides.

Therapeutic resistance represents a formidable bottleneck in colorectal cancer (CRC) management, severely limiting the clinical efficacy of both conventional regimens and novel immunotherapies. Emerging evidence underscores tertiary lymphoid structure (TLS) - ectopic lymphoid aggregates newly sculpted within the tumor microenvironment (TME) - as pivotal orchestrators of localized antitumor immunity and prime therapeutic targets to circumvent resistance. Distinct from secondary lymphoid organs, TLS facilitate in situ immune cell priming, clonal expansion, and functional differentiation, thereby sustaining a robust adaptive immune response. This review systematically dissects the cellular architecture, maturation dynamics, and spatial heterogeneity of TLS in primary and metastatic CRC, clarifying how these attributes dictate clinical outcomes and treatment responsiveness. Mechanistically, we delineate the molecular networks driving TLS neogenesis, encompassing chemokine cascades, tumor necrosis factor (TNF) superfamily signaling, and interactive crosstalk with the gut microbiota. Furthermore, we summarize cutting-edge preclinical strategies - including stimulator of interferon genes agonists, bio-nanovaccines, and hypofractionated radiotherapy combinations - engineered to trigger TLS neogenesis, promote germinal center (GC) maturation, and enrich stem-like effector tumor-infiltrating lymphocytes (TILs) to convert immunologically “cold” tumors into “hot” niches. By integrating mechanistic paradigms with clinical applications, this review provides a definitive framework for leveraging TLS targeting as a transformative modality to modulate drug resistance and optimize therapeutic trajectories in CRC; the graphical abstract is shown below.

Background: Psychoneurological symptom clusters (PNSCs) are common in patients with ovarian cancer and are associated with reduced quality of life, treatment interruption, and poor prognosis. However, effective interventions for PNSCs remain limited. Traditional Chinese medicine may provide comprehensive benefits for symptom management. Objective: This study aims to evaluate the efficacy and safety of the TiaoShenZhiAi (TSZA) regimen in alleviating PNSCs in patients with ovarian cancer and to assess its effects on quality of life and survival outcomes. Methods: A total of 316 patients with ovarian cancer aged 18 to 70 years with PNSCs will be included and randomly divided into 2 parallel groups. Both groups will receive standard treatment for ovarian cancer as the basic treatment. The intervention group will receive the TSZA regimen, that is, Compound Ciwujia Granules (containing Acanthopanax senticosus and Schisandra chinensis) combined with psychological intervention. The control group will receive a low-dose active control (simulated Compound Ciwujia Granules) combined with psychological intervention. The primary outcome is the remission rate of PNSCs at 3 months. The secondary outcome measures include the Pittsburgh Sleep Quality Index, the Patient Health Questionnaire-9, the Generalized Anxiety Disorder-7 scale, the revised Piper Fatigue Scale, the European Organization for Research and Treatment of Cancer Quality of Life Questionnaire - Core 30 Quality of Life Scale, the traditional Chinese medicine syndrome scale, sleep quality, sleep diary, and the 1-year survival analysis. In addition, this study also includes a series of exploratory indicators (including functional magnetic resonance imaging, biomarkers of peripheral blood and tumor tissue, proportion of immune cells, cytokine levels, hypothalamic-pituitary-adrenal axis function, and immune gene expression analysis) and safety indicators (including vital signs, liver and kidney function, and electrocardiogram). The study outcomes will be evaluated based on different indicators during the treatment period (baseline and the 1st, 2nd, and 3rd mo of enrollment) and the follow-up period (the 6th, 9th, and 12th mo of enrollment). Data analysis will be conducted using R (version 4.5.3) software. A one-sided P value of <.03 will be considered statistically significant. Results: This study is designed to enroll a total of 316 participants. Participant enrollment is set to commence in October 2025, with no recruitment having occurred as of April 2026. The recruitment period will extend until September 2028 or until the target enrollment is met. Data analysis is scheduled for November 2028, with submission of the trial results to a peer-reviewed journal anticipated by May 2029. Conclusions: This study will evaluate the efficacy of the TSZA regimen in managing PNSCs in patients with ovarian cancer and generate clinical evidence for a new therapeutic option that improves quality of life and alleviates the symptom burden.

Colorectal cancer (CRC) exhibits pronounced biological diversity, a feature increasingly attributed to alterations in cellular metabolic reprograms. Serine metabolism supports nucleotide synthesis, redox balance, and epigenetic regulation via one-carbon metabolism, yet its role in shaping the tumor cellular interactions and immune landscape at single-cell level remains unclear. Single-cell transcriptomic profiles (GSE284449) were jointly analyzed with bulk expression datasets (GSE271719), together with Mendelian randomization (MR)-based inference, to dissect serine metabolism in CRC. High-serine-metabolism (HSM) cell populations were identified using scMetabolism, and robust marker genes were selected through Lasso, random forest, XGBoost, and SVM-RFE machine learning approaches. MR was applied to evaluate causal associations with CRC risk. Functional validation included IHC, qRT-PCR, western blot, and LC–MS metabolomics, while CellChat analysis characterized HSM cell interactions with immune and stromal cells. ZBTB21 and TRPM2 were identified as core regulators of HSM cells, with ZBTB21 predominantly expressed in monocytes and pro-B cells. MR analysis suggested a potential inverse association between genetically predicted ZBTB21 expression and CRC risk, indicating that ZBTB21 may exert context-dependent effects at the population level. Cell–cell communication analysis suggested that HSM monocytes may interact with fibroblasts through signaling pathways including the MIF–SPP1 axis; however, these interactions are computationally inferred and require further experimental validation. Functional assays showed that ZBTB21 overexpression upregulated PHGDH and SHMT1, increased intracellular NADPH/NADP⁺ ratios, and influenced monocyte-related phenotypes. ZBTB21 levels were markedly increased in CRC samples relative to matched non-tumorous mucosal tissues. At single-cell resolution, ZBTB21 emerges as a metabolic regulator that strengthens serine biosynthesis and redox homeostasis. While integrative analyses suggest a potential link between ZBTB21-associated metabolic states and immune interactions, further experimental validation is required to establish causal relationships. This integrative framework connects genetic causality, metabolism, and immune interactions, providing mechanistic insights and potential strategies for metabolic-targeted therapies in CRC. Trial registration: Not applicable.

Background:Serum magnesium plays a critical role in modulating immune responses and enhancing anti-tumor immunity in cancer patients. However, its predictive significance in relation to progression-free survival (PFS) and disease control rate (DCR) among hepatocellular carcinoma (HCC) patients undergoing first-line immunotherapy remains uninvestigated. Methods:This retrospective study analyzed HCC patients treated with immune checkpoint inhibitors (ICIs) in a derivation cohort and an independent validation cohort. Patients were enrolled based on predefined inclusion criteria. The primary endpoint was to assess the association between baseline serum magnesium levels prior to immunotherapy initiation and the disease control rate. The optimal cut-off value of serum magnesium for predicting disease control was determined using receiver operating characteristic (ROC) curve analysis. PFS was evaluated using the Kaplan-Meier method. Results:A total of 110 hepatocellular carcinoma patients treated with ICIs were enrolled in the derivation cohort. The DCR was 78.18%. ROC analysis identified a baseline serum magnesium level ≥0.785 mmol/L as predictive of disease control, with a sensitivity of 89.4% and specificity of 87.5% (AUC 0.869). Patients with baseline Mg2+ ≥0.785mmol/L demonstrated significantly longer median PFS compared to those with baseline Mg2+<0.785mmol/L (12.83 months vs. 3.45 months, P< 0.001, hazard ratio: 0.269, 95% confidence interval: 0.165-0.438). The DCR was 30% in the low-Mg2+ group and 96.25% in the high-Mg2+ group. Subgroup analyses revealed that patients with macrovascular invasion or extrahepatic metastasis, as well as those without macrovascular invasion or extrahepatic metastases in the high-Mg2+ group exhibited significantly longer median PFS compared to those in the low-Mg2+ group. Both univariate and multivariate analyses confirmed that serum magnesium level is an independent predictive factor of PFS for patients receiving immunotherapy. In the validation cohort (n=48), patients with Mg2+ ≥0.785 mmol/L showed significantly longer median PFS (17.13 months vs. 5.20 months, P = 0.037, hazard ratio: 0.304, 95% confidence interval: 0.131-0.701) and higher DCR compared to those with Mg2+<0.785mmol/L. Conclusion:Elevated serum magnesium levels prior to first-line immunotherapy are associated with improved clinical outcomes in HCC. Serum magnesium demonstrates significant predictive value and could serve as a cost-effective, accessible biomarker for guiding immunotherapy strategies in HCC patients.

Hashimoto’s thyroiditis is associated with reduced lymph node metastasis in papillary thyroid carcinoma, suggesting that the autoimmune microenvironment may influence tumor progression. We utilized single-cell RNA sequencing and intratumoral 16S ribosomal RNA gene sequencing to investigate malignant epithelial cell states and microbial features in PTC with and without Hashimoto’s thyroiditis. Tumors with Hashimoto’s thyroiditis exhibited malignant epithelial populations enriched for innate immune signaling programs and showed increased intratumoral microbial diversity with reduced abundance of several Gram-negative bacteria, particularly Pseudomonas. Within the Hashimoto’s thyroiditis subgroup, lymph node metastasis was associated with increased Gram-negative bacterial abundance. Functional experiments further showed that Pseudomonas-derived lipopolysaccharide enhanced thyroid cancer cell migration in vitro and in vivo. These findings suggest that Hashimoto’s thyroiditis reshapes both epithelial transcriptional states and the intratumoral microbial environment, potentially contributing to a less aggressive tumor phenotype.

The simultaneous modulation of oxidative stress and autophagy represents a potential therapeutic strategy for amyotrophic lateral sclerosis (ALS), yet agents capable of coordinately regulating both processes remain scarce. The Keap1‑Nrf2‑ARE pathway serves as a critical hub linking redox homeostasis and autophagic regulation, making it an attractive target for ALS intervention. JWA is a stress‑responsive protein involved in cellular protection against oxidative injury, and its neuroprotective effects have been shown to depend on activation of the MEK/ERK‑Nrf2 axis. JP1 is a functional oligopeptide derived from the JWA protein that has been engineered to cross the blood–brain barrier and specifically target integrin αVβ3. Based on the link between JWA and Nrf2 signaling, we hypothesized that JP1 activates the Keap1‑Nrf2‑ARE pathway to coordinate antioxidant defense and autophagic clearance. Here, we evaluated this hypothesis in the SOD1‑G93A mouse model, a well‑established transgenic model of familial ALS, and elucidated the underlying mechanisms. We evaluated the efficacy of JP1 in the SOD1-G93A mice model using behavioral phenotyping and survival analysis. The coordinated mechanism was investigated in spinal cord tissues by profiling the Keap1-Nrf2-ARE pathway and oxidative stress, quantifying autophagic flux (by Western blotting and transmission electron microscopy) and neuronal apoptosis, and evaluating histology (by Nissl staining and immunofluorescence). Integrated transcriptomic and proteomic analyses further elucidated the global molecular landscape underlying the therapeutic effects of JP1. JP1 treatment ameliorated motor deficits and extended survival in SOD1-G93A mice without adversely affecting liver or kidney function. JP1 crossed the blood–brain barrier, targeted motor neurons expressing integrin αVβ3, and activated the ERK pathway. This promoted Keap1/Cul3 degradation and Nrf2 nuclear translocation, thereby activating the Keap1-Nrf2-ARE pathway to alleviate oxidative stress. Concurrently, JP1 restored autophagic flux, increased autophagic activity, attenuated motor neuron injury, suppressed neuronal apoptosis, and preserved neuronal structural integrity. The Nrf2 inhibitor ML385 reversed the protective effects of JP1 on survival, motor function, autophagy, oxidative stress, and neuronal apoptosis, which confirms that JP1 acts via the Nrf2 pathway. JP1 acts as a promising coordinator of antioxidant and autophagic processes by targeting the Keap1-Nrf2-ARE pathway, thus highlighting its therapeutic potential for ALS.

OBJECTIVE:Small cell carcinoma of the cervix (SCCC) is a rare and highly aggressive malignancy with limited evidence to guide surgical management. We therefore aimed to evaluate the prognostic value of regional lymph node (LN) surgery and LN status in patients with SCCC undergoing primary tumor resection. METHODS:In this retrospective cohort study, data were obtained from the Surveillance, Epidemiology, and End Results database (January 1, 2000 to December 31, 2021). A total of 204 patients with histologically confirmed SCCC who underwent primary tumor resection were included and classified according to receipt of regional LN surgery. An independent validation cohort with 121 patients was retrospectively identified from our center (January 1, 2013 to December 31, 2020). Kaplan-Meier curves, multivariable Cox proportional hazards models, and stratified subgroup analyses were performed. RESULTS:Of the 204 included patients, 141 (69.1%) underwent regional LN surgery and 63 (30.9%) did not. Baseline demographic and clinicopathologic characteristics were similar between groups. In patients with International Federation of Gynecology and Obstetrics (FIGO) stage I-IVA disease, regional LN surgery was not associated with improved cancer-specific survival (CSS) (adjusted hazard ratio [HR], 1.16; 95% confidence interval [CI], 0.52-2.56; P = 0.722), and LN-negative status was not prognostic. Among those with stage IVB disease, LN surgery was associated with significantly better survival (adjusted HR, 0.24; 95% CI, 0.08-0.77; P = 0.016). In the independent validation cohort, LN status was not significantly associated with recurrence-free survival (HR, 1.163; 95% CI, 0.37-3.61; P = 0.794) or CSS (HR, 1.39; 95% CI, 0.44-4.40; P = 0.575). CONCLUSION:Regional LN surgery did not improve survival among patients with FIGO stage I-IVA SCCC, and LN status alone was not an independent prognostic indicator. However, LN surgery may confer potential benefit in FIGO stage IVB disease. These findings suggest that LN dissection should be considered carefully and highlight the need for individualized surgical decision-making and prospective validation.

In the original publication [...].

Small cell carcinoma of cervix (SCCC) was a highly aggressive tumor with dismal prognosis. Current treatment strategies manifested poor survival outcomes and novel treatment options were needed exploration. We aimed to investigate several prognostic biomarkers for SCCC and conducted a novel risk-score system to predict cancer specific survival (CSS) in early-stage SCCC. Seven cell-cycle proteins were detected by immunohistochemistry in 88 SCCCs. Univariate and multivariate analysis were performed to identify prognostic proteins and establish a predicting model. Total patients were divided into two groups by the median risk-score: the high-risk group and the low-risk group. Logistic regression and Wilcoxon test were used to investigate the association between clinical variables and the risk-score system. Seven cell cycle proteins were overexpressed in SCCC. The expression of CDC20, MAD2L1, MCM2 and BUBR1 were correlated to survival outcomes with P < 0.05. A novel risk-score system consisting of CDC20, MAD2L1 and BUBR1 was significantly an independent prognostic factor for CSS and the high-risk group possessed worse survival (P < 0.001). The c-indexes for clinical model, risk-score system and the combined model were 0.668, 0.718 and 0.727, respectively. The AUCs for these three models were 0.730, 0.775 and 0.823, respectively. Furthermore, we discovered that patients with high-risk scores were inclined to possessing older age, parametrial invasion and higher FIGO stage (IIA vs IA/IB) with P < 0.05. This risk-score system consisting of CDC20, MAD2L1 and BUBR1 presented good discrimination and predictability for SCCC. Novel biomarkers in this study might have some merits in providing guidance of novel treatment strategies for SCCC.

Ubiquitin conjugating enzyme UBE2S is an important enzyme in ubiquitin mediated proteolysis. The mechanisms of UBE2S in the colorectal cancer (CRC) proliferation need further exploration for new ubiquitination-related drugs exploitation. Firstly, we performed immunohistochemistry of UBE2S protein on CRC and adjacent tissues and found that UBE2S protein expressed higher in adjacent tissues than in tumor. TCGA and GTEx databases revealed a positive correlation between UBE2S expression and overall survival rate in CRC (P < 0.05). Cell proliferation experiments on HCT116 and RKO cell lines invalidated that UBE2S overexpression inhibited CRC cell proliferation and progression from G0/1 phase to G2 phase while knocking down UBE2S enhanced it. Mechanistically, combined with enrichment analysis based on RNA sequencing, it was identified molecularly that UBE2S simultaneously inhibited the activation of MAPK/ERK and PI3K/AKT pathways through promoting the ubiquitin-mediated degradation of TRAF6 protein, an upstream molecule to activate the both pathways, which can be blocked by inhibitor MG132. Animal experiments demonstrated that knocking down UBE2S resulted in larger transplanted tumor volumes than the control group, while overexpressing UBE2S resulted in the smaller ones in vivo. Summarily, we firstly identified that the UBE2S inhibited CRC proliferation by negatively regulating MAPK/ERK and PI3K/AKT/mTOR pathways through ubiquitinating TRAF6 protein.

BACKGROUND:Tertiary lymphoid structures (TLS) are emerging biomarkers for enhanced anti-tumoral immunity and treatment efficacy in colorectal cancer (CRC). This study aimed to develop a CRC-specific TLS signature and evaluate its predictive roles in survival prognosis and therapeutic responses to chemotherapy, and PD-1 blockade. MATERIALS AND METHODS:Transcriptomic data from TCGA database were analyzed to establish a TLS signature. Survival and treatment responses were validated in multiple cohorts, including TCGA cohort, RJ cohort (neoadjuvant FOLFOX, n = 16), CHcohort (neoadjuvant sintilimab, n = 17), and several GEO datasets. Genetic mutations, tumor microenvironment (TME) features, and its spatial interactions were assessed via maftools, Bayesprism, CIBERSORT, IBOR, TIMER 2.0, NicheNet analysis and multiplex immunofluorescence. Tumor-immune cells co-cultured miniPDX/PDO models were used to validate PD-1 blockade in MSS CRC. RESULTS:In TCGA cohort, patients in TLS_high group exhibited significantly longer overall survival ( P = 0.008) and demonstrated a higher proportion of complete response/partial response (CR/PR) ( P = 0.017). Further analyses on TME revealed that tumor mutation burden was positively correlated with TLS signature scores ( R = 0.31, P < 0.001). High TLS scores depicted a higher proportion of CD8 + T cells, B cells, and dendritic cells, increased activation of anti-tumoral signaling pathways, enhanced cytokines secretion, and higher immune checkpoints expression, including CTLA4, LAG3, and TIM3. Sensitive group in RJ cohort and clinical CR group in CH cohort scored higher of TLS signature ( P < 0.05), and depicted activation of signaling pathways related to TLS formation. Mature TLS and enhanced intercellular receptor-ligand interactions were identified. miniPDX/PDO models confirmed TLS-driven PD-1 blockade sensitivity in MSS CRC. CONCLUSIONS:The CRC-specific TLS signature identifies patients with favorable prognosis and enhanced responses to chemotherapy and PD-1 blockade, including MSS CRC resistant to immune checkpoint blockade. TLS abundance reflects an immunogenic TME, offering a biomarker for personalized treatment strategies.

e15624 Background: Surgical resection remains the cornerstone treatment for locally advanced rectal cancer. However, for patients at high risk of recurrence, even with resectable tumors, the prognosis is still unfavorable. PD-1 inhibitors have shown promise in treating dMMR/MSI-H advanced metastatic rectal cancer, achieving an objective response rate of 30%-40% in patients who have previously failed treatments. This clinical trial aims to evaluate the efficacy and safety of combining tislelizumab (an anti-PD-1 antibody) with concurrent chemoradiotherapy as neoadjuvant therapy for patients with locally advanced rectal cancer. Methods: In this phase II clinical trial (ChiCTR2400094540), patients with resectable rectal cancer were divided into two parallel cohorts (1:1) based on different neoadjuvant treatment protocols. Patients in group A received chemoradiotherapy combined with tislelizumab, while patients in Group B received chemoradiotherapy alone. All patients were scheduled to undergo radical surgery after completing neoadjuvant treatment. The primary endpoint was the pathological complete response (pCR) rate. In addition, whole-exome, transcriptome, and immune repertoire sequencing technologies were utilized to explore potential biomarkers and changes in the immune microenvironment associated with the prognosis of neoadjuvant treatment in rectal cancer patients. Results: As of November 30, 2024, a total of 28 patients were enrolled in this study, with 16 in the experimental group and 12 in the control group. Of these, 26 patients completed surgical treatment, while 2 patients in the control group did not undergo surgery. There were notable differences in tumor regression grade (TRG) and lymph node remnants between the two groups. Due to the small sample size, no significant difference was observed in pCR rates between the groups (31.2% vs 30%). However, a difference was noted in major pathological response (MPR) rates (68.8% vs 30%). Tumor-associated macrophage M2 may influence prognosis. Both a low positive rate of tumor parenchyma and a high positive rate of tumor interstitial tissue were indicative of a better prognosis, although no significant difference was found. Conclusions: For patients with locally advanced rectal cancer, the addition of tislelizumab to neoadjuvant chemoradiotherapy appears to increase the rates of pathological complete response (pCR) and major pathological response (MPR), although further research is needed to confirm these findings. Clinical trial information: ChiCTR2400094540 .