Temporary ileostomy is commonly performed to protect rectal anastomosis after rectal surgery, yet the optimal timing of reversal remains unsettled. This study compared postoperative outcomes among early, intermediate, and late ileostomy closure and assessed the safety of early closure in selected patients. PubMed, Web of Science, and Embase were systematically searched from inception to June 21, 2026. Randomized controlled trials comparing different timing strategies for ileostomy closure were included. Closure timing was categorized according to the reported median or mean interval from primary surgery to reversal as early (≤ 35 days), intermediate (> 35 and ≤ 120 days), and late (> 120 days) based on the median or mean interval. The primary outcomes were total postoperative complications and major complications. A Bayesian network meta-analysis was performed. Ten randomized trials involving 871 patients (316 early, 312 intermediate, and 243 late) were included. Total postoperative complications were comparable among the three groups. Compared with intermediate closure, early closure was associated with higher risks of major complications (odds ratio [OR] 5.30, 95

The clinical application of conventional chemotherapeutic drugs is often limited by systemic toxicity and insufficient tumor accumulation. Herein, we report a dual-responsive polymeric nanocarrier, PLGA-C8E nanoparticles (NPs), designed to enhance drug delivery by integrating morphology engineering with stimulus-responsive release. The NPs were fabricated via a phase separation-driven assembly process, resulting in a distinctive mushroom-like morphology composed of a poly(lactic-co-glycolic acid) (PLGA) vesicular domain and a Cys-8E-rich solid domain. Owing to the presence of ester and disulfide bonds, the nanocarriers exhibited pH- and glutathione-responsive drug release behavior. In vitro studies demonstrated good biocompatibility and efficient cellular uptake. In vivo biodistribution analysis revealed prolonged blood circulation and enhanced tumor-associated accumulation. Consequently, PTX@PLGA-C8E NPs achieved significantly improved tumor growth inhibition while maintaining favorable systemic biosafety. These results highlighted the critical role of non-spherical, mushroom-like morphology in modulating in vivo behavior and therapeutic efficacy, demonstrated the potential of shape-engineered nanocarriers in advancing nanomedicine design, which may offer valuable insights for the development of next-generation translational drug delivery systems.

The neutrophil percentage-to-albumin ratio (NPAR) is an emerging indicator of inflammation that has been associated with the prognosis of various diseases, including hypertension, diabetes, and cardiovascular disease. NPAR is calculated by multiplying the neutrophil percentage by 100 and dividing it by the albumin value. This study aimed to evaluate the predictive value of NPAR for all-cause mortality in anemic patients. We employed Kaplan–Meier analysis, multiple regression models, restricted cubic splines (RCS), and threshold effect analysis to explore these associations. This study included 3,258 anemic individuals. The relationship between NPAR and all-cause mortality was evaluated using Kaplan–Meier analysis and multiple regression models. RCS curves were employed to assess potential nonlinear relationships, while threshold effect analysis was used to identify breakpoints in the association between NPAR and mortality risk. Subgroup analyses were conducted to examine variations in this relationship across different population strata. The study population included 3,258 anemic patients, with a mean follow-up period of 94.3 months. During follow-up, 904 participants (27.75

Food intake has a significant effect on systemic physiology. The growth, development and transmission of malaria-causing Plasmodium parasites rely on carbohydrate utilization, and parasites must rapidly adapt to fluctuating host glucose concentrations throughout their life cycle. However, the precise process by which Plasmodium parasites quickly react and adapt to glucose stress, in general, remains elusive. The goal of this investigation was to determine whether glucose starvation affects the RNA secondary structure of parasites. In this study, we applied different short-term glucose stresses on parasites to determine the effect of host-derived glucose nutrients on the dynamics of parasite development and RNA secondary structures. We found that the S-type rRNA expression level in the low glucose-treated group was higher than that in the 2 mg/mL control group. The RNA-seq results indicated that glucose significantly affects the expression of genes involved in RNA-related processes, antigenic variation, transport processes, stimulus response, and DNA-related activities. Metabolomics studies also revealed that several types of sugar compounds exhibit a decrease in metabolic activity when subjected to a reduction in glucose supply. Via analysis of three glucose conditions, we identified the RNA secondary structures of more than 2,721 transcripts throughout the intraerythrocytic trophozoite stage. We discovered a positive correlation between mRNA structural flexibility (icSHAPE scores) and transcript abundance (RPKM values), demonstrating a direct association between RNA structural openness and steady-state mRNA levels. Our results revealed structural alterations in the entire transcriptome, including glucose transporters, metabolic enzymes, and the erythrocyte invasion protein family. These genes were mainly enriched in response to stimuli or preferential biological processes. The total number of genes with changes in icSHAPE scores in the UTRs was significantly greater for the 3′ UTRs than the 5′ UTRs. Furthermore, by comparing icSHAPE reactivity profiles across glucose conditions, we identified candidate cis-acting RNA motifs predominantly located within UTRs, whose structural flexibility changed rapidly in response to glucose fluctuation. We term these glucose-responsive RNA motifs, as they constitute promising regulatory elements for future research into post-transcriptional glucose sensing mechanisms. Our findings clarify glucose stress-triggered remodeling of in vivo RNA secondary structure during the P. falciparum intraerythrocytic stage, reveal the functional relevance of structural dynamics in parasite environmental adaptation, and lay a foundational resource for future nutrient stress and post-transcriptional regulatory research. These findings show that glucose, the primary carbon source for parasite survival, modulates RNA secondary structural dynamics via nutrient-sensing cascades to modulate parasite proliferation and adaptive development, which offers clues for developing novel antimalarial therapies.

TPS271 Background: Locally advanced colorectal cancer (LACRC) is recommended to receive neoadjuvant chemotherapy(NCT) by the NCCN guidelines before surgery. NCT might reduce tumor size, achieve downstaging of the tumor and lymph nodes, improve R0 resection rate, enhance prognosis or even watch-and-wait. Previous studies have shown a 2.07% retrieval rate of lymph nodes beyond 5 cm along the bowel for cases classified as T≤3, with an even lower incidence of positive lymph nodes. Retrospective cohort comparison at our center showed that reducing the extent of bowel resection (3 cm of distal margin and 5 cm of proximal margin) after NCT did not affect the lymph node retrieval and the 1-year DFS. Extending the resection length of the bowel or expanding the scope of regional mesenteric lymph node dissection beyond 5 cm does not improve staging accuracy. Instead, it might increase the risk of complications . For LACRC with ycT≤3N0M0 (stage IIA or lower) after NCT, the optimal extent of bowel resection remains a subject for further investigation. Methods: This is a prospective, multicenter, randomized phase III-controlled trial conducted in collaboration with the Sixth Affiliated Hospital of Sun Yat-sen University and multiple domestic colorectal cancer centers. Patients with upper rectal or rectosigmoid junction tumors at a clinical stage of cT3-4N+M0 will be enrolled. For patients with pMMR/MSS status: Tumor downstaged to T≤3N0M0 after NCT or total neoadjuvant therapy (TNT). Chemotherapy regimens include, but are not limited to, FOLFOX6, CAPOX, or FOLFOXIRI. For patients with dMMR/MSI-H: Tumor downstaged to T≤3N0M0 after immunotherapy. The trial will compare conventional resection margins with reduced margins in patients who have met the ycT ≤3N0M0 regression criteria following NCT or TNT. Reduced resection margins refer to the surgical resection region including 5 cm bowel proximal to the tumor, 3 cm bowel distal to the tumor and lymph node dissection in the corresponding mesentery. Conventional resection margins refer to the surgical resection region including 10 cm bowel proximal to the tumor, 5 cm bowel distal to the tumor, and lymph node dissection in the corresponding mesentery. The goal of this clinical trial is to learn evaluate the safety and efficacy of reduced surgical resection margins in patients with ycT≤3N0M0 regression following NCT. The primary research objective is 3-year disease-free survival (DFS). The secondary research objectives include R0 resection rate, number of lymph nodes dissected, surgical safety metrics and other survival outcomes. This study plans to enroll 874 patients, with 10 patients enrolled as of August 30th, 2025. Registration with ClinicalTrials.gov (NCT 07038122) has been completed. Clinical trial information: NCT 07038122 .

Malaria remains a critical parasitic disease in tropical regions, with environmental temperature significantly influencing the development and transmission of Plasmodium falciparum. While low temperature triggers gametocyte differentiation in mosquito, the molecular mechanisms underlying temperature-responsive chromatin and transcriptional dynamics in asexual stages (ring and trophozoite) remain unclear. This study integrates Assay for Transposase-Accessible Chromatin with Sequencing (ATAC-seq) and RNA-sequencing (RNA-seq) to characterize genome-wide chromatin accessibility and gene expression profiles in P. falciparum under human body temperature (37 °C) and mosquito-mimicking temperature (26 °C). Synchronized ring (45 h post-invasion) and trophozoite (70 h post-invasion) stages were subjected to temperature treatments (37 °C versus 26 °C). ATAC-seq was used to identify accessible chromatin regions, RNA-seq analyzed differentially expressed genes (DEGs), and quantitative real-time polymerase chain reaction (qPCR) validated key gene expression changes. Low temperatures exerts a profound impact on the activation and expression of sexual-stage-specific genes in P. falciparum and induced 1083 differentially accessible regions (DARs) in the ring stage, including 1081 gains and only 2 losses, which were enriched primarily in promoter regions (≤ 3 kb upstream of transcription start sites), whereas no significant DARs were detected in the trophozoite stage, indicating stage-specific sensitivity to temperature. Functional analyses revealed DAR-associated genes enriched in host cell membrane interactions, antigenic variation, and pathways such as symbiont-mediated perturbation of host erythrocyte aggregation, with the identification of temperature-responsive transcription factor motifs (e.g., DEAR-3, ERF096). Integration of ATAC-seq and RNA-seq revealed a positive correlation between chromatin accessibility and gene expression, with 41 genes exhibiting concordant DAR-DEG changes, suggesting that dynamic chromatin remodeling regulates temperature-responsive transcription. Low temperature selectively modulates chromatin accessibility and gene expression in the ring stage, while trophozoites show no chromatin reconfiguration, highlighting stage-specific thermal sensitivity. This study represents the first integrative analysis of ATAC-seq and RNA-seq data from P. falciparum under low temperature stress, identifying critical temperature-responsive regulatory regions, providing insights into the parasite’s environmental adaptation and a foundational resource of temperature-responsive regulatory regions, whose future functional validation could inform the development of novel, chromatin-targeted antimalarial strategies.

TPS3646 Background: Immunotherapy has shown promising therapeutic effects in mismatch repair-deficient or microsatellite instability-high (dMMR/MSI-H) colorectal cancer (CRC). However, for patients with mismatch repair-proficient or microsatellite stable (pMMR/MSS) CRC, the efficacy of single-agent PD-1 monoclonal antibody remains limited. Previous studies reported that combining anti-angiogenic drugs with PD-1 monoclonal antibody might improve the efficacy of immunotherapy. Our BASKETII study (NCT04895137) demonstrated that the neoadjuvant therapy regimen of mFOLFOX6 combined with Bevacizumab and sintilimab significantly enhanced the immunotherapy sensitivity of pMMR/MSS locally advanced CRC (LACRC), resulting in improved pathological complete response (pCR) rates and higher R0 resection rates. Methods: BASKETIII is a multicenter, randomized controlled, phase III study with a parallel design conducted in China. This trial aims to evaluate whether the neoadjuvant therapy regimen of mFOLFOX6 combined with Bevacizumab and sintilimab can further improve survival outcomes, and maintain the higher pCR rate and acceptable safety profile compared to mFOLFOX6 in pMMR/MSS LACRC patients. Eligible participants will be randomly assigned in a 1:1 ratio to either the experimental group or the control group. Participants in the experimental group will receive the neoadjuvant therapy regimen of mFOLFOX6 + Bevacizumab + sintilimab. The first five doses will follow the mFOLFOX6 combined with Bevacizumab and sintilimab regimen, and the sixth dose will receive only mFOLFOX6 and sintilimab but without Bevacizumab, in order to avoid delay of surgery. Participants in the control group will receive the neoadjuvant therapy regimen of mFOLFOX6 alone. Participants in both groups will undergo radical surgical treatment after neoadjuvant therapies. Participants who achieve pCR based on postoperative pathology will be regularly followed up. Participants who do not achieve pCR will receive adjuvant therapy with a maxim of six doses and will be regular followed up after the final dose of adjuvant therapy. The primary outcome of this study is to evaluate the 3-year disease-free survival (DFS). The key inclusion criteria include histologically confirmed adenocarcinoma of the colon or upper rectum; tumor biopsy immunohistochemical identified pMMR or MSS identified through next-generation sequencing or polymerase chain reaction; Clinical staging of cT4NxM0. The main exclusion criteria include evidence of distant metastasis beyond the pelvic region; history of pelvic or abdominal radiotherapy; multiple CRC or multiple primary tumors; history of immunotherapy and other malignancies within the past 5 years. A total of 122 patients are planned to be enrolled in this study. This study is registered with ClinicalTrials.gov (NCT06791512) and is recruiting. Clinical trial information: NCT06791512 .

Chemotherapy drug 5-Fluorouracil (5-FU) is a major treatment for many cancers; however, its efficacy is limited by chemoresistance. Here, we investigate the resistance mechanisms to 5-FU and reversal strategies in lung and breast cancer cells. Using multiple 5-FU-resistant lung cancer and breast cancer cell models, we reveal differential cellular and molecular features of 5-FU resistance between different cancer types. We further unravel the implications of immune-related processes, NOTCH and WNT signaling with 5-FU resistance. In lung cancer, the activation of WNT/β-catenin signaling promotes the resistance and blocking this signaling re-sensitizes resistant cells to 5-FU treatment. Our study not only reveals differential features and mechanisms underlying 5-FU resistance across different cancers, but also suggests potential strategies against such resistance.

IntroductionSophorae flavescentis (kushen) preparations are widely used to control malignant pleural effusion (MPE) through intrapleural perfusion.ObjectivesThis analysis aims to verify the therapeutic values of perfusion with kushen preparations for controlling MPE, reveal the optimal treatment plan, suitable population, and usage, and to demonstrate their clinical effectiveness and safety.MethodsWe performed and reported this systematic review/meta-analysis (PROSPERO: CRD42023430139) following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. All randomized controlled trials (RCTs) concerning perfusion with kushen preparation for MPE were collected from Chinese and English databases. We clustered all eligible studies into multiple homogeneous treatment units, assessed their methodological quality using a RoB 2, pooled the data from each unit, and summarized the quality of the evidence.ResultsWe included 83 RCTs reporting three types of kushen preparation: compound kushen injection (CKI), kang’ai injection, and matrine injection. All trials were clustered into perfusion with CKI alone or with the addition of sclerosants, kang’ai, or matrine-plus platinum for controlling MPE. Compared with cisplatin alone, perfusion with CKI alone displayed a similar complete response, pleurodesis failure, and pleural progression (odds ratios =1.10, 95% CI 0.76 to 1.60; 0.80, 0.56 to 1.14; 0.63, 0.33 to 1.21). Of 14 homogeneous treatment plans, perfusion with CKI and cisplatin significantly improved the complete response (2.71, 2.30 to 3.19) and showed low pleurodesis failure (0.26, 0.22 to 0.32), pleural progression (0.22, 0.14 to 0.36), myelosuppression (0.34, 0.24 to 0.47), neutropenia (0.35, 0.26 to 0.46), gastrointestinal reaction (0.36, 0.29 to 0.44), hepatorenal toxicity (0.42, 0.28 to 0.63 and 0.32, 0.24 to 0.44), and fever (0.50, 0.30 to 0.82). These results were moderate quality (⊕⊕⊕Ο) supported by firm or conclusive information. Additionally, perfusion with kang’ai or matrine and cisplatin also improved the complete response (3.04, 1.76 to 5.26 and 1.87, 1.26 to 2.78) and displayed low pleurodesis failure (0.23, 0.14 to 0.41 and 0.27, 0.17 to 0.44). The results were moderate to low quality (⊕⊕⊕Ο to ⊕⊕ΟΟ).ConclusionCurrent moderate evidence demonstrates that CKI may be an effective palliative intervention for MPE which, combined with cisplatin, may be an optimal treatment plan. Kang’ai or matrine may be other potential choices.Systematic Review Registration:https://www.crd.york.ac.uk/PROSPERO/view/CRD42023430139

The formation of robust vascular systems within voluminous scaffolds remains a formidable barrier in the realm of tissue engineering. There is a growing interest in the integration of biomaterial scaffolds with multiple physical and chemical stimuli to augment the process of vascularization. This study aims to investigate the combined impact of macroporous structures and vascular endothelial growth factor (VEGF) on cell migration and vascularization. Heparinized hyaluronic acid (HepHA) macroporous hydrogels with differing pore sizes, composed by methacrylated hyaluronic acid (HAMA) and methacrylated heparin (HepMA), were fabricated by a gelatin microspheres (GMS) template leaching method. After characterization of their physical properties, VEGF was immobilized on the HepHA hydrogels. The in vitro release study indicated that the HepHA hydrogels can provide sustained release of VEGF. Subsequently, cells migration of human umbilical vein endothelial (HUVECs) assessment indicated that HUVECs cultured on VEGF-loaded HepHA hydrogels with larger pores (VEGF@HepHA250) migrated the furthest. Finally, the hydrogels were implanted and evaluated using a dorsal subcutaneous model. The histological analyses conducted in vivo were consistent with the in vitro results, VEGF@HepHA250 hydrogels exhibited the most pronounced vascularization four weeks post-implantation, indicating that hydrogels with expanded pores and an enriched VEGF promoted angiogenesis within the hydrogels. This study sheds light on the synergistic effects of VEGF release on 3D cell migration and vascularization within hydrogels of differing pore sizes, thus providing novel insights into the strategic design and fabrication of tissue-engineered scaffolds that are amenable to vascularization.

OBJECTIVES:To evaluate the efficacy and safety of unrelated donor allogeneic hematopoietic stem cell transplantation (URD-HSCT) combined with umbilical cord blood infusion in the treatment of malignant hematological diseases. METHODS:The clinical data of 11 patients with malignant hematological diseases who received unrelated allogeneic hematopoietic stem cell transplantation combined with umbilical cord blood infusion were retrospectively summarized. RESULTS:After treatment, the median times for neutrophil engraftment were 11 days (11-15 days), and 13 days (12-22 days) for platelets engraftment. Four patients (36%) developed grade II-IV acute graft versus host disease (aGVHD), which mainly affected gastrointestinal tract, the skin, and the liver. One patient (9%) had grade IV aGVHD affecting gastrointestinal tract. Chronic graft versus host disease (cGVHD) occurred in three patients (27%), mainly affecting the skin and liver. The median follow-up time was 26 months (6-50 months). The 3-year overall survival (OS) and GVHD relapse-free survival rates were 80.8% and 60.6%, respectively. None of the 11 patients experienced a relapse by the end of the observation. CONCLUSION:Despite the small number of cases, our data show that URD-HSCT in combination with unrelated cord blood transfusion is a safe and effective therapy for patients with intermediate- to high-risk malignant hematological diseases.

Autoimmune uveitis (AU) is a sight-threatening ocular autoimmune disorder that often manifests as retinal vasculitis. Increased neutrophil infiltration around retinal vessels has been reported during the progression of AU, while how they function is not fully recognized. Neutrophil extracellular traps (NETs), produced by activated neutrophils, have been suggested to be detrimental in autoimmune diseases. Here, we found that NETs were elevated in patients with active AU, and this was verified in an experimental AU (EAU) mouse model. Depletion of neutrophils or degradation of NETs with deoxyribonuclease-I (DNase I) could decrease CD4+ effector T cell (Teff) infiltration in retina and spleen to alleviate EAU. Moreover, we found that the expression of adhesion molecules, selectin, and antigen-presenting molecules was elevated in EAU retina and in retinal microvascular endothelial cells (RMECs) cocultured with NETs. The stimulated RMECs further facilitated CD4+ T cell adhesion, activation, and differentiation into Teffs. Mechanistically, NETs trigger RMEC activation by hastening cell senescence through the cyclic GMPAMP synthase (cGAS)/stimulator of interferon genes (STING) pathway. Slowing down senescence or inhibiting the cGAS/STING pathway in RMECs reduces the activation and differentiation of CD4+ T cells. These results suggest a deleterious role of NETs in AU. Targeting NETs would offer an effective therapeutic method.

Compared to traditional antibiotics, which often contribute to antibiotic resistance, probiotics and their metabolites have garnered increasing attention for their potential to improve the wound microenvironment. Among these, reuterin, a metabolite produced by Limosilactobacillus reuteri, exhibits notable antioxidant and antimicrobial activities, making it particularly advantageous for medical applications. Inspired by the structural similarity of reuterin to conventional chemical crosslinkers, we explored its dual role for the first time-as both a bioactive component and a crosslinking agent-by combining it with chitosan to develop a novel therapeutic hydrogel, eliminating the need for additional drug loading. At an optimal reuterin-to-chitosan molar ratio of 1:2, the hydrogel demonstrated outstanding performance. Cytotoxicity assays confirmed its safety, with a hemolysis rate below 2 %, indicating excellent biocompatibility. Remarkably, the hydrogel achieved an impressive 99 % antibacterial efficacy and over 60 % DPPH radical scavenging capacity in vitro. In a rat model of infected wounds, the hydrogel exhibited acid-responsive release of reuterin, effectively suppressing bacterial proliferation. These combined effects significantly accelerated wound healing, demonstrating an approximately 10 % improvement in healing rates compared to commercial controls. In conclusion, the reuterin-crosslinked chitosan hydrogel represents a promising, safer and more effective alternative to traditional chemical crosslinked hydrogels for clinical wound management.

Mannatide is an alpha-hemolytic streptococci bio-product. We conducted a comprehensive clustered systematic review and meta-analysis to reveal the clinical values of Mannatide for controlling malignant pleural effusion (MPE), identify the optimal therapeutic regimens, indications and usage, and assess their clinical effectiveness and safety. We collected all eligible studies from both Chinese and English databases and clustered them into homogeneous comparisons or therapeutic regimens. We evaluated the risk of bias in eligible studies using RoB 2, pooled the data from each regimen using Review Manager 5.4, and summarized the quality of evidence. Seventy-eight studies met the eligibility criteria. Among six comparisons of perfusion with sclerosant alone, Mannatide showed only better complete response of MPE and lower pleurodesis failure and disease progression than interleukin-2. Among twelve combination therapeutic regimens, Mannatide and cisplatin might significantly improve the complete response (2.46, 2.07 to 2.91) and result in a low pleurodesis failure (0.28, 0.23 to 0.33) and disease progression (0.32, 0.21 to 0.48). For MPE with primary treatment, moderate to massive effusion, Karnofsky Performance Status ≥ 40 to 60 or anticipated survival time ≥ two to three months, perfusion with Mannatide at 20 to 50 mg/time, once per week and two to four times and cisplatin at 30 to 60 mg/time might achieve ideal clinical response. Most outcomes were moderate to low quality. Mannatide may be a potential pleural sclerosant, which combination with cisplatin may represent a preferred therapeutic approach with a good clinical response and safety in controlling MPE. PROSPERO CRD42022338798.

Genomic analyses have revolutionized risk stratification and clinical management of pediatric B-cell acute lymphoblastic leukemia (B-ALL), but these methods are time-consuming and costly. To address this gap, we employed highly sensitive and specific Olink proteomics analysis of bone marrow plasma samples from pediatric B-ALL patients and B-ALL patients with complete molecular remission (CMR) as controls. Fifty pediatric B-ALL patients and 43 controls were randomly divided into a “discovery” cohort (30 B-ALL patients, 26 CMR controls), and a “validation” cohort (20 B-ALL patients and 17 CMR controls). The Olink Target 96 Oncology Response protein panel was used. Normalized log2-scale protein expression values revealed 37 differentially expressed proteins (DEPs) between pediatric B-ALL and CMR control groups. LASSO regression analysis showed nine of these DEPs, including CXCL13, NCR1, ADA, IL6, HO-1, CCL3, CCL4, CD27, and ADGRG1, to be significant in differentiating between patients with B-ALL and CMR controls (validation AUC = 0.98). We further strengthened the study by validating key Olink-identified proteins using independent assays, including ELISA in patient bone marrow plasma and immunohistochemistry in patient-derived xenograft tissues, which confirmed some Olink proteomic findings, such as CCL3, CCL4, and CD27. In addition, we found that higher levels of DEPs, such as CD27, TNF, CCL3, CCL4, IL12RB1, PDCD1, and GZMB, in patients with genetic alterations associated with poor prognosis. Our Olink proteomics analysis identified nine key proteins that were differentially expressed between the pediatric B-ALL and control groups, which may contribute to the diagnosis and/or risk stratification of pediatric B-ALL.

3601 Background: Neoadjuvant immunotherapy provides considerable advantages for patients with dMMR/MSI-H rectal cancer. However, treatment strategies in real-world settings vary depending on tumor characteristics, economic conditions, and the choices made by physicians and patients. Methods: We screened more than 10,000 rectal cancer cases from seven large medical college-affiliated hospitals. We used the Kaplan-Meier curve to compare survival and progression, applied Cox regression to analyze impact factors, and examined tumor regression grades with chi-square analysis. Results: From March 2010 to April 2024, 502 patients were enrolled and diagnosed with dMMR/MSI-H rectal adenocarcinoma through immunohistochemistry or PCR. 100 patients underwent neoadjuvant immunotherapy, demonstrating a 96.34% 5-year overall survival (95% CI: 86.08-99.08%), and 90.74% 5-year disease-free survival (95% CI: 74.67-96.82%). This indicated a 16.50% enhancement in overall survival (p = 0.042) and a 16.87% increase in disease-free survival (p = 0.002) compared to conventional chemoradiotherapy (5-year OS: 79.84%, 95% CI: 71.69-85.87%; 5-year DFS: 73.87%, 95% CI: 65.15-80.73%). Neoadjuvant immunotherapy demonstrated significant superiority in tumor regression (p < 0.0001), however, the combination with chemotherapy did not enhance the effect (p = 0.622), and varying chemotherapeutic agents did not improve tumor regression in conventional chemoradiotherapy either. Additionally, elevated serum CEA levels were associated with an increased risk of both death and disease progression. Patients with advanced age, lower clinical stages, and fewer risk factors were more likely to undergo direct surgical resection. Conclusions: Neoadjuvant immunotherapy is advantageous for dMMR/MSI-H rectal cancer patients, as it leads to better tumor regression and enhanced disease control.

Plasma proteins have been reported as predictors and potential targets for reducing colorectal cancer (CRC) risk. However, their potential roles in CRC prognosis remain unexplored. We measured plasma levels of 367 neuro-related proteins in CRC patients from the West China Hospital (WCH) cohort (N = 150, median follow-up = 46.72 months) via proximity extension assay. The least absolute shrinkage and selection operator penalized Cox regression identified five overall survival (OS)-and eleven disease-free survival-associated proteins, and the multiprotein signature for OS prediction was then validated in the UK Biobank (UKB) cohort (N = 1133). To overcome possible effects from confounders, we then employed Mendelian randomization analysis leveraging protein quantitative trait loci to investigate associations between genetically determined protein concentration and OS and cancer-specific survival of CRC in the UKB. We found that multiprotein signature developed in the WCH cohort (c-index = 0.784, 95% CI = 0.713-0.855) showed significant discriminative ability in the external UKB cohort (c-index = 0.616, 95% CI = 0.559-0.673). A significant association between genetically determined PD-L1 and OS (P = 0.043, HR = 1.53, 95% CI = 1.01-2.29) was observed, although we did not find strong evidence for colocalization. Additionally, single-cell and spatial transcriptome analyses illustrated PD-L1 expression localized predominantly to epithelial cells and immune cells (especially myeloid cells) in CRC tissue. The potential interactions of identified proteins were evaluated in the STRING database. Druggability evaluation also supported PD-L1 as a potential therapeutic target for CRC. Taken together, this study established multiprotein signatures for CRC prognosis and identified plasma PD-L1 as a possible biomarker and therapeutic target.

Purpose:The purpose of this study was to explore the underlying mechanism that Th17-like T follicular helper cells (Tfh) orchestrated by STING signaling have a pathogenic role in experimental autoimmune uveitis (EAU). Methods:The differences of transcriptome and gene ontology (GO) pathway of Tfh between EAU and control mice were analyzed by single-cell RNA sequence (scRNA-seq) and bulk RNA sequence. Additionally, draining lymph nodes (DLNs) were extracted to verify the expression of IL-17A and IFN-γ in Tfh from EAU and control mice by flow cytometry. Then, the scRNA-seq and flow cytometry were used to explore the different proportion of Tfh between STING deficiency (Sting-/-) mice and wild type (WT) mice. In vitro, naïve CD4+ T cells were isolated from Sting-/- mice and WT mice to induce the Tfh under the induction condition. In addition, flow cytometry was used to detect the different induction ratio and the IL-17A expression between 2 groups of naïve CD4+ T cells. Results:Compared with control mice, marked increase of Tfh was observed in EAU, accompanied by elevated levels of Th1 and Th17 cells. Moreover, Th17-related genes, such as Rorc, Il22, Il23r, Il17a, and Il17f, and the corresponding GO pathways were upregulated in Tfh from EAU. The scRNA-seq showed that a higher proportion of Tfh was observed in the DLNs from Sting-/- mice than WT mice, which was verified by flow cytometry. When STING was knocked out, the Tfh was characterized with upregulated Th17-related phenotype in vivo, and there was a higher induction ratio of Tfh whose IL-17A expression was significantly increased in vitro. Notably, the STING expression of CD4+ T cells was downregulated in the EAU. STING-deficient EAU mice displayed more severe retinal inflammation, characterized by massive infiltration of CD4+ T cells, including Th1 and Th17 subsets. Importantly, treatment with a STING agonist alleviated inflammation of EAU. Conclusions:Th17-like Tfh cells play a pathogenic role in the EAU. STING deficiency promotes the differentiation and phenotypic transformation of Th17-like Tfh cells, exacerbating the inflammatory response in EAU. These findings highlight the potential of targeting STING to modulate Tfh cells as a therapeutic strategy for uveitis.