Background The nectin cell adhesion molecule 4 (NECTIN4) has been implicated in tumor progression and immune evasion, yet its role and translational targeted imaging potential in lung cancer remain unclear. Therefore, this study aims to elucidate the significance of NECTIN4 by integrating multi-omics analyses, and to evaluate the diagnostic efficacy of the NECTIN4-targeted PET/CT imaging in lung cancer. Methods Transcriptomic and proteomic datasets from The Cancer Genome Atlas (TCGA), Genotype-Tissue Expression (GTEx), Gene Expression Omnibus (GEO) and other bioinformatic tools were used to characterize NECTIN4 expression, genomic alterations, epigenetic regulation, and prognostic relevance in lung cancer. Subsequently, in a prospective clinical cohort study involving 20 patients with suspected primary lung cancer, paired PET/CT imaging using Ga-68-N188 and F-18-FDG was conducted. Diagnostic performances were assessed by quantitatively comparing the tumor-to-blood pool ratio between malignant and inflammatory lesions. Results Bioinformatics analyses indicated that NECTIN4 was significantly upregulated across multiple cancer types and correlated with genomic instability and poor prognosis in non-small cell lung cancer (NSCLC). NECTIN4 expression was positively associated with DNA methyltransferases and RNA modifications, suggesting that it may be regulated by epigenetic and post-transcriptional. As for NECTIN4-targeted imaging, Ga-6(8)-N188 PET/CT exhibited superior specificity (100% vs. 50%) and comparable sensitivity (87.5% vs. 93.8%) to & sup1;F-8-FDG PET/CT in differentiating malignant from inflammatory lung lesions, but with lower sensitivity (42.2% vs. 100.0%) for detecting lymph node metastases and fewer identified distant metastatic lesions (21 vs. 51). Conclusion Integrated bioinformatics analyses prove that overexpression of NECTIN4 is associated with occurrence and progression of lung cancer. Further preliminary clinical translation study suggests the potential of NECTIN4-targeted radiotracer Ga-6(8)-N188 to aid in the differential diagnosis of lung cancer, highlighting a promising clinical application warrants further validation.

Abstract Background Endothelial cell-specific molecule-1 (ESM1) is a secreted protein involved in a variety of tumors. Nevertheless, the role of ESM1 in oral squamous cell carcinoma (OSCC) is still unclear. This study aimed to elucidate the function and mechanisms of ESM1 in OSCC. Methods Differences in ESM1 expression in OSCC and para-carcinoma tissues were detected by immunohistochemistry, and an ESM1 overexpression human umbilical vein endothelial cell (HUVEC) model was constructed. CCK-8, EdU staining, and colony formation assays were employed to evaluate cell proliferation capacity, while Transwell and scratch assays were performed to assess cell invasion and migration capabilities. Quantitative polymerase chain reaction and western blotting assays were used to analyze the changes in endothelial–mesenchymal transition (EndMT) markers, and bioinformatics was conducted to verify the role of the NOTCH1/DLL4 pathway. The effects of ESM1 on the malignant phenotype of tumor cells and epithelial–mesenchymal transition (EMT) were detected by a co-culture model of HUVEC and OSCC cells. Results ESM1 was significantly upregulated in OSCC tissues and correlated with advanced T-stage, N-stage, and low immune score. ESM1 was secreted by endothelial cells, and overexpression of ESM1 enhanced the proliferation, migration, and invasion ability of HUVEC, as well as the activation of EndMT. Mechanistically, ESM1 induced EndMT through activating the NOTCH1/DLL4 pathway. Co-culture experiments showed that overexpression of ESM1 in HUVEC significantly promoted the proliferation, migration, invasion, and EMT processes of OSCC cells. Conclusion ESM1 mediates EndMT in endothelial cells through the NOTCH1/DLL4 pathway and promotes EMT and the malignant phenotype of OSCC cells through paracrine effects, suggesting its potential as a therapeutic target for OSCC.

Background Musculoskeletal sarcomas are rare neoplasms originating from mesenchymal cells and are generally malignant. The presence and persistence of circulating tumor cells (CTCs) are increasingly investigated for their potential significance in tumor evaluation. Currently, various methods have been reported to efficiently extract and examine CTCs in patients’ circulation. However, sarcomas lack classical biomarkers and the ultimate method which could both realize efficiency and activity preservation in sarcomas is still yet to be investigated. Here, we designed a combined strategy of Microporous Filter Membrane and Immunofluorescence (MPMF-IF) for both detection and characterization of CTCs in patients with sarcomas. Methods We conducted numerical experiments to evaluate capture efficiency and purity of the present method through testing sarcoma cell lines and blood samples. Subsequently, the characteristics of the strategy were further investigated in vivo. Also, the device’s performance was validated in patients of different clinical stages. Results Satisfying results of high capture efficiency, purity and specificity at the same time preserving cellular activity were confirmed in experiments in vitro. Experiments in vivo certified that the device could functionally analyze tumor burden. In clinical samples, the significant difference was detected between metastatic patients and non-metastatic patients and the results illustrated that CTC counts by MPMF-IF could predict the prognosis of patients. Conclusions Our MPMF-IF method for the detection and characterization of CTCs enables satisfying accuracy and efficiency. A great potential could be expected in further applying MPMF-IF to predict the prognosis and evaluate the therapeutic effect for patients with musculoskeletal sarcomas.

Whole lung lavage (WLL) remains the standard treatment for patients with pulmonary alveolar proteinosis (PAP). However, WLL procedures are not fully standardized, and there is no global consensus on the best approach, such as performing one-session versus two-session WLL. This study aimed to assess and compare the safety and effectiveness of these two methods in a real-world clinical environment. In this retrospective cohort study, we included patients with PAP who underwent WLL at a tertiary hospital in China from September 2009 to May 2024. Patients were categorized into one-session and two-session WLL groups. The primary outcome was the incidence of WLL-related complications. Secondary outcomes included improvements in ΔPaO2, procedure duration, the rate of extubation within 24 hours, length of hospital stay, and hospitalization costs. A total of 26 patients with PAP underwent 37 WLL procedures, with 15 procedures in the one-session group (15 patients) and 22 procedures in the two-session group (11 patients). The incidence of WLL-related complications, including acid-base disorders, desaturation, and pleural effusions, did not significantly differ between the groups. However, compared to the two-session WLL group, the one-session group demonstrated significantly greater improvement in ΔPaO2 (14.20 [2.6, 29.2] mmHg vs 2.50 [-3.2, 5.45] mmHg, p=0.015) and shorter hospital stay (10 [7, 14] days vs 18 [10, 26] days, p=0.020). No significant differences were observed in the procedure duration, extubation rate within 24 hours, or hospitalization costs between the groups. For patients with PAP, one-session WLL offers comparable safety to two-session WLL, with significantly greater improvement in ΔPaO2 and shorter hospital stays, without increasing hospitalization costs. These results suggest that adopting one-session WLL may optimize patient outcomes and resource utilization in clinical practice.

R-loop accumulation has emerged as a critical factor that induces DNA damage and compromises genomic integrity. However, the regulatory mechanisms governing the R-loop-induced DNA damage remain unclear. Here, FTSJ3 was determined to be a pivotal regulator of R-loop homeostasis and genomic stability. We demonstrated that FTSJ3 was specifically recruited to R-loop structures, where it prevented DNA damage by suppressing excessive R-loop formation. FTSJ3 expression was significantly upregulated in multiple cancer types, and its elevated expression levels correlated with unfavorable survival in patients with lung adenocarcinoma (LUAD). FTSJ3 depletion increased R-loop-dependent DNA damage. Inhibiting FTSJ3 expression sensitized lung cancer cells to cisplatin both in vitro and in vivo. FTSJ3 could be a genome guardian that limits R-loop-associated damage, suggesting its potential role as a cancer intervention therapeutic target and a predictive biomarker for chemotherapy responsiveness.

In vitro models coupled with multimodal approaches are needed to dissect the dynamic response of local tumor immune microenvironment (TIME) to immunotherapy. Here the patient-derived primary lung cancer organoids (pLCOs) are generated by isolating tumor cell clusters, including the infiltrated immune cells. A function-associated single-cell RNA sequencing (FascRNA-seq) platform allowing both phenotypic evaluation and scRNA-seq at single-organoid level is developed to dissect the TIME of individual pLCOs. The analysis of 171 individual pLCOs derived from seven patients reveals that pLCOs retain the TIME heterogeneity in the parenchyma of parental tumor tissues, providing models with identical genetic background but various TIME. Linking the scRNA-seq data of individual pLCOs with their responses to anti-PD-1 (αPD-1) immune checkpoint blockade (ICB) allows to confirm the central role of CD8+ T cells in anti-tumor immunity, to identify potential tumor-reactive T cells with a set of 10 genes, and to unravel the factors regulating T cell activity, including CD99 gene. In summary, the study constructs a joint phenotypic and transcriptomic FascRNA-seq platform to dissect the dynamic response of local TIME under ICB treatment, providing a promising approach to evaluate novel immunotherapies and to understand the underlying molecular mechanisms.

Purpose There were fewer data to guide the application of enhanced recovery after surgery (ERAS) theory into sacral tumour surgery. In the present study, we were aiming to describe a multidisciplinary program of ERAS and evaluate the availability in sacral tumour surgery. Methods This was a prospective study of patients with sacral tumour between March 2021 and September 2021 at a single centre. We proposed a multidisciplinary program of ERAS for pre-admission, preoperative, intraoperative, postoperative, and post-discharge clinical care which positively influenced outcomes of patients with sacral tumour. All patients were prospectively assigned into two groups, ERAS group in which patients received ERAS protocols ( n = 63), No-ERAS group in which patients had conventional clinical pathways ( n = 62). Patient data were collected which included demographics, preoperative preparation, detailed information of surgical procedure, 60-day reoperation rate, 60-day readmission, postoperative length of stay (PLOS), time to first ambulation and flatus after surgery, time to removal of last drainage tube, and visual analogue scale (VAS) score at first ambulation and discharge. Complications referred to ones that occurred within 60 days after surgery. The above parameters were compared between ERAS group and No-ERAS group. Results Time to first ambulation after surgery in ERAS group (mean 20.9 h) was significantly shorter than that in No-ERAS group (mean 104.3 ho). Meanwhile, time to first flatus after surgery in ERAS group (mean 26.7 h) was also significantly shorter than that in No-ERAS group (mean 37.3 h). Patients in ERAS group had statistically shorter PLOS (10.7 days) as compared to that in No-ERAS group (13.8 days). In ERAS group, 19 of 63 patients (30.2%) were discharged within seven days after surgery as compared to seven of 62 patients (11.3%) in No-ERAS group. VAS score at first ambulation in ERAS group was not obviously higher than that in No-ERAS group though the time of first ambulation in ERAS group was statistically earlier than one in No-ERAS group. Furthermore, VAS score at discharge in ERAS group was significantly lower than that in No-ERAS group. The rate of postoperative incision necrosis was 6.3% (4/63) in ERAS group and 8.1% (5/62) in No-ERAS group and all of these nine patients underwent reoperation before discharge. The difference was not statistically significant in the wound complication of incision necrosis and 60-day reoperation rate. Only one readmission occurred in No-ERAS group due to the surgical site infection and also there was no significant difference of 60-day readmission rate between these two groups. Furthermore, there was no statistical difference of complications of femoral artery thrombosis and rectal rupture between ERAS group and No-ERAS group. Conclusions Our proposed ERAS pathway for sacral tumour surgery and early walking facilitate safe and prompt discharge. ERAS protocols of sacral tumour surgery could decrease PLOS without significantly increasing postoperative complications, 60-day readmission rate and 60-day reoperation rate. The application of ERAS pathway in the field of sacral tumour surgery should have personalized feature with regard to resection type.

Figure S1. (A) Strategy for plasma detection of oncogenic mutations. The exonic positions and nature of the hotspot mutations are shown. Two sets of bidirectional reverse primers (arrows) were positioned to robust sequences on either side of each mutation site to target and amplify each allele by inverse PCR. For detection of ALK fusions, targeting primers were progressively spaced across exon 19, intron 19, and exon 20, spanning the known ALK breakpoint region. (B) Hot Spot Gene Mutations and Diagnostic cSMART Primers. Figure S2. Comparison of the distributions of driver mutations identified in 26 matched tDNA and plasma ctDNA samples. Thirty-two mutations were detected in the plasma at time A, including 14 TP53 mutations, 8 EGFR mutations, 6 PIK3CA mutations, 3 KRAS mutations and 1 ALK mutation. Five patients had two mutations simultaneously in the plasma ctDNA: case 3 had PIK3CA and EGFR mutations; case 6 had EGFR and TP53 mutations; case 15 had two different TP53 mutations, namely, a base substitution mutation in exon 5 and a base substitution mutation in exon 8; case 17 had EGFR and TP53 mutations; and case 20 had KRAS and PIK3CA mutations simultaneously. A single mutation was detected in the other 21 cases. For the corresponding tumor tissues, a total of 29 gene mutations were detected: 14 TP53 mutations, 8 EGFR mutations, 5 PIK3CA mutations and 2 KRAS mutation. More than one mutation was detected in the tumors of 5 patients, which was completely consistent with the ctDNA results. (A) Numbers of gene mutations in tDNA vs. plasma ctDNA from matched sample pairs. (B) The inner circle shows the distribution of gene mutations, and the outer ring shows specific alterations in amino acids. Figure S3. Excluded two positive plasma A patients with CHIP validated by normal lung tissue and white blood cell. Figure S4. The plasma ctDNA concentration at each time point during the perioperative period (from time A to time P2). The X-axis represents the logarithmic value of time. Figure S5. Longitudinal ctDNA profiles of non-relapse or non-progression cases. Red curve represents the fluctuation of ctDNA concentration over time. The X-axis represents the days of post-operation. The Y-axis represents the ctDNA MAF (%). In this figure, rectangular frames represent the treatment that patients have received, as shown at the bottom, green frame: surgical treatment; yellow frame: chemotherapy; blue frame: radiotherapy and red frame: target therapy. Figure S6. The RFS (left) and OS (right）between different ctDNA statuses at time P3 (Three cases with no blood samples at this time point were excluded). Figure S7. The RFS between patients who received postoperative therapy or not based on different strategy. (A) Patients with stage II, III and above lung cancer according to TNM stage in our study. (B) Patients with positive MRD detection in our study. (C) Patient with positive ctDNA at time P2.

BACKGROUND:We compare the application of intravenous indocyanine green (ICG) fluorescence imaging in lung cancer with near-infrared-I (NIR-I) and near-infrared-II (NIR-II) windows.METHODS:From March to December 2022, we enrolled patients who received an intravenous injection of ICG (5 mg/kg) 1 day before the planned lung cancer surgery. The lung cancer nodules were imaged by NIR-I/II fluorescence imaging systems, and the tumor-to-normal-tissue ratio (TNR) was calculated. In addition, the fluorescence intensity and signal-to-background ratio (SBR) of capillary glass tubes containing ICG covered with different thicknesses of lung tissue were measured by NIR-I/II fluorescence imaging systems.RESULTS:In this study, 102 patients were enrolled, and the mean age was 59.9 ± 9.2 years. A total of 96 (94.1%) and 98 (96.1%) lung nodules were successfully imaged with NIR-I and NIR-II fluorescence, and the TNR of NIR-II was significantly higher than that of NIR-I (3.9 ± 1.3 versus 2.4 ± 0.6, P < 0.001). In multiple linear regression, solid nodules (P < 0.001) and squamous cell carcinoma (P < 0.001) were independent predictors of a higher TNR of NIR-I/II. When capillary glass tubes were covered with lung tissue whose thickness was more than 2 mm, the fluorescence intensity and the SBR of NIR-II were significantly higher than those of NIR-I.CONCLUSIONS:We verified the feasibility of NIR-II fluorescence imaging in intravenous ICG lung cancer imaging for the first time. NIR-II fluorescence can improve the TNR and penetration depth of lung cancer with promising clinical prospects.

Our study aimed to investigate whether serum total IgE and blood eosinophils were associated with radiological features of bronchiectasis in a Chinese cohort. We retrospectively enrolled bronchiectasis patients who visited Peking University Third Hospital from Jan 1st, 2012 to Oct 7th, 2021. The clinical, laboratory and chest CT characteristics were analyzed in association with serum total IgE level and blood eosinophil count. A total of 125 bronchiectasis patients were enrolled, with 50.4

In the era of histopathology-based diagnosis, the discrimination between multiple lung cancers (MLCs) poses significant uncertainties and has thus become a clinical dilemma. However, recent significant advances and increased application of molecular technologies in clonal relatedness assessment have led to more precision in distinguishing between multiple primary lung cancers (MPLCs) and intrapulmonary metastasis (IPMs). This review summarizes recent advances in the molecular identification of MLCs and compares various methods based on somatic mutations, chromosome alterations, microRNAs, and tumor microenvironment markers. The paper also discusses current challenges at the forefront of genomics-based discrimination, including the selection of detection technology, application of next-generation sequencing, and intratumoral heterogeneity (ITH). In summary, this paper highlights an entrance into the primary stage of molecule-based diagnostics.

Circulating tumor DNA (ctDNA) is DNA fragment shed from tumor cells and can be isolated from peripheral blood. ctDNA carries genomic information of cancer and is the most widely utilized indicator in liquid biopsy. The application of ctDNA is gaining traction due to its reproducible, non-invasive, and easy-to-obtain characteristics. With the development of detection approaches, ctDNA plays an important role in the management of patients with cancers. In this review, we summarize the basic ctDNA detection measurements and review its application in early screening, prognosis evaluation after the surgery, and efficacy prediction of different therapies in non-small cell lung cancer.

All data were from the cited references.