Overexpression of SMAD7 improves the function of EGFR-targeted human CAR-T cells against non-small-cell lung cancer.

BACKGROUND AND OBJECTIVE: Recent advancements in immunotherapy led to the development of Chimeric antigen receptor (CAR) T-cell therapy. CAR-T cell therapy in non-small cell lung cancer (NSCLC) is hindered by overexpression of transforming growth factor (TGFß) in the cancer cells that have a negative regulatory role on T-cells activity. This study characterized CAR-T with overexpression of mothers against decapentaplegic homologue 7 (SMAD), a negative regulator of TGFß downstream signalling. METHODS: We have generated three types of CAR-T: epidermal growth factor receptor (EGFR)-CAR-T, EGFR-dominant-negative TGFbeta receptor 2 (DNR)-CAR-T, and EGFR-SMAD7-CAR-T by transducing human T-cells with the lentivirus constructs. We characterized the proliferation, expression of proinflammatory cytokines, activation profile, and lysis capacity in co-cultures with A549 lung carcinoma cells with and without TGFß neutralizing antibodies. We also tested the therapeutic potential of EGFR-SMAD7-CAR-T in the A549 cells tumour-bearing mice model. RESULTS: Both EGFR-DNR-CAR-T and EGFR-SMAD7-CAR-T demonstrated a higher proliferation rate and lysis capacity to A549 than traditional EGFR-CAR-T. Neutralization of TGFß by the antibodies resulted in increased performance of EGFR-CAR-T. In vivo, both EGFR-DNR-CAR-T and EGFR-SMAD7-CAR-T resulted in complete tumour resorption by day 20, whereas conventional CAR-T only has a partial effect. CONCLUSION: We demonstrated the high efficacy and resistance to negative TGFß regulation of EGFR-SMAD7-CAR-T comparable with EGFR-DNR-CAR-T and without the systemic effect of TGFß inhibition.

Prediction of distant metastasis and specific survival prediction of small intestine cancer patients with metastasis: A population-based study.

BACKGROUND: Small intestine cancer (SIC) is difficult to diagnose early and presents a poor prognosis due to distant metastasis. This study aimed to develop nomograms for diagnosing and assessing the prognosis of SIC with distant metastasis. METHODS: Patients diagnosed with SIC between 2010 and 2015 were included from the Surveillance, Epidemiology and End Results database. Univariate and multifactor analysis determined independent risk factors for distant metastasis and prognostic factors for overall and cancer-specific survival. We then constructed the corresponding three nomograms and assessed the diagnostic accuracy of the nomograms by net reclassification improvement, receiver operating characteristic curves and calibration curves, assessed the clinical utility by decision curve analysis. RESULTS: The cohort consisted of 6697 patients, of whom 1299 had distant metastasis at diagnosis. Tstage, Nstage, age, tumor size, grade, and histological type were independent risk factors for distant metastasis. Age, histological type, T stage, N stage, grade, tumor size, whether receiving surgery, number of lymph nodes removed, and the presence of bone or lung metastases were predictors of both overall survival and cancer-specific survival. The nomograms showed excellent accuracy in predicting distant metastasis and prognosis. CONCLUSION: Nomograms were developed and validated for SIC patients with distant metastasis, aiding physicians in making rational and personalized clinical decisions.

Application of single-port laparoscopic retrograde gastric mobilization during McKeown esophagectomy for esophageal cancer.

BACKGROUND: As a novel alternative to the conventional minimally invasive esophagectomy (MIE) to treat esophageal cancer, single-port laparoscopic retrograde three-step gastric mobilization (SLRM) for esophageal reconstruction during MIE to treat esophageal cancer was attempted in our department. The aim of the present study was to explore the preliminary clinical outcomes and feasibility of this innovative surgery. METHODS: From March 2020 to November 2021, patients undergoing SLRM combined with four-port thoracoscopic McKeown esophagectomy for their esophageal cancers were reviewed. Gastric mobilization with abdominal lymph node dissection was performed through SLRM. The clinical characteristics and short-term outcomes were analyzed retrospectively. RESULTS: A total of 120 patients underwent R0 resection without conversion to open surgery. The mean times needed for the thoracic part, abdominal part, and total operation were 43 ± 6 min, 60 ± 18 min, and 230 ± 20 min, respectively. The numbers of mediastinal and abdominal lymph nodes harvested were 13.2 ± 2.7 and 10.2 ± 2.5, respectively. Postoperative pneumonia was encountered in 10 (8.3%) patients. Anastomotic leakage occurred in 3 (2.5%) cases. Temporary vocal cord paralysis was reported in 20 (16.6%) cases. The mean length of hospital stay was 8.5 ± 4.6 days. CONCLUSIONS: The SLRM is a technically feasible and safe treatment for patients with esophageal cancer. It can be considered an alternative method for patients, especially for the ones with obesity and gastric distension.

Intelligent Diagnosis of Multiple Peripheral Retinal Lesions in Ultra-widefield Fundus Images Based on Deep Learning.

INTRODUCTION: Compared with traditional fundus examination techniques, ultra-widefield fundus (UWF) images provide 200° panoramic images of the retina, which allows better detection of peripheral retinal lesions. The advent of UWF provides effective solutions only for detection but still lacks efficient diagnostic capabilities. This study proposed a retinal lesion detection model to automatically locate and identify six relatively typical and high-incidence peripheral retinal lesions from UWF images which will enable early screening and rapid diagnosis. METHODS: A total of 24,602 augmented ultra-widefield fundus images with labels corresponding to 6 peripheral retinal lesions and normal manifestation labelled by 5 ophthalmologists were included in this study. An object detection model named You Only Look Once X (YOLOX) was modified and trained to locate and classify the six peripheral retinal lesions including rhegmatogenous retinal detachment (RRD), retinal breaks (RB), white without pressure (WWOP), cystic retinal tuft (CRT), lattice degeneration (LD), and paving-stone degeneration (PSD). We applied coordinate attention block and generalized intersection over union (GIOU) loss to YOLOX and evaluated it for accuracy, sensitivity, specificity, precision, F1 score, and average precision (AP). This model was able to show the exact location and saliency map of the retinal lesions detected by the model thus contributing to efficient screening and diagnosis. RESULTS: The model reached an average accuracy of 96.64%, sensitivity of 87.97%, specificity of 98.04%, precision of 87.01%, F1 score of 87.39%, and mAP of 86.03% on test dataset 1 including 248 UWF images and reached an average accuracy of 95.04%, sensitivity of 83.90%, specificity of 96.70%, precision of 78.73%, F1 score of 81.96%, and mAP of 80.59% on external test dataset 2 including 586 UWF images, showing this system performs well in distinguishing the six peripheral retinal lesions. CONCLUSION: Focusing on peripheral retinal lesions, this work proposed a deep learning model, which automatically recognized multiple peripheral retinal lesions from UWF images and localized exact positions of lesions. Therefore, it has certain potential for early screening and intelligent diagnosis of peripheral retinal lesions.

Utility of single-port laparoscopic retrograde gastric mobilization during McKeown esophagectomy for esophageal cancer: a 2-year experience with 120 cases in a single institution.

Background: As a novel alternative to the conventional minimally invasive esophagectomy (MIE), more minimally invasive single-port laparoscopic retrograde 3-step gastric mobilization (SLRM) for esophageal reconstruction during MIE to treat esophageal cancer was attempted by our department. This study explored the preliminary clinical outcomes and feasibility of this innovative surgery. Methods: The data of 120 patients who had undergone SLRM combined with 4-port thoracoscopic McKeown esophagectomy for their esophageal cancers from March 2020 to November 2021 were reviewed. Gastric mobilization with abdominal lymph node dissection was performed via SLRM. The clinical characteristics and short-term outcomes were retrospectively analyzed. The data of operating time, blood loss, harvested lymph nodes, postoperative hospital stay and complications are presented as the mean and standard deviation. Results: A total of 120 patients underwent R0 resection without conversion to open surgery. The mean times for the thoracic procedure, abdominal procedure, and total operation were 43±6, 60±18, and 195±20 min, respectively. The numbers of mediastinal and abdominal lymph nodes harvested were 13.2±2.7, and 10.2±2.5, respectively. Postoperative pneumonia occurred in 10 patients (8.3%). Anastomotic leakage occurred in 3 patients (2.5%). Temporary vocal cord paralysis was reported in 20 patients (16.6%). The mean length of hospital stay was 8.5±4.6 days. Conclusions: SLRM is a technically feasible and safe treatment for patients with esophageal cancer. It can be considered an alternative method for patients, especially those with obesity and gastric distension.

ERO1L promotes the proliferation and metastasis of lung adenocarcinoma via the Wnt2/ß-catenin signaling pathway.

PURPOSE: This study aimed to explore the role and underlying mechanism of action of Endoplasmic reticulum oxidoreductin-1 L (ERO1L) in lung adenocarcinoma (LUAD). MATERIALS AND METHODS: The Gene expression profiling interactive analysis database was used to analyze the expression of ERO1L in LUAD cases. The expression of ERO1L and Wnt2 in LUAD tissue was evaluated using immunohistochemistry. We also used western blotting to assess the expression of ERO1L or Wnt2 and the phosphorylation of ß-catenin in LUAD cell lines. Plasmid transfection and small interfering RNA were used for overexpression and knockdown of ERO1L in LUAD cells, respectively. The proliferation, invasion and migration of LUAD cells were analyzed using cell viability, Transwell invasion and wound healing assays. The growth of LUAD tumors in animal models was assessed using tumor xenograft experiments. RESULTS: This study revealed that elevated ERO1L expression was associated with a poor prognosis in LUAD patients. In addition, ERO1L expression was significantly associated with lymph-node metastasis, TNM stage and tumor size. The expression of Wnt2 was positively associated with ERO1L expression in LUAD tissue samples and cell lines. ERO1L overexpression upregulated the expression of Wnt2 and ß-catenin phosphorylation in vitro. Additionally, ERO1L was essential for the ubiquitination of Wnt2. Last, ERO1L promoted the proliferation and metastasis of LUAD via the Wnt2 signaling pathway in vitro and in vivo. CONCLUSION: These findings suggest that ERO1L was highly expressed in LUAD tissue, and it promoted the proliferation and metastasis of LUAD by activating the Wnt2/ß-catenin signaling pathway.

A lightweight network for automatic thyroid nodules location and recognition with high speed and accuracy in ultrasound images.

BACKGROUND: The intelligent diagnosis of thyroid nodules in ultrasound image is an important research issue. Automatically locating the region of interest (ROI) of thyroid nodules and providing pre-diagnosis results can help doctors to diagnose faster and more accurate. OBJECTIVES: This study aims to propose a model, which can detect multiple nodules stably and accurately in order to avoid missed detection and misjudgment. In addition, the detection speed of the model needs to be fast for real-time diagnosis in ultrasound images. METHODS: Based on the object detection technology, we propose an accurate, robust and high-speed network with multiscale fusion strategy called Efficient-YOLO, which can realize the localization and recognition of nodules at the same time. Finally, multiple metrics are used to measure the diagnostic ability of the model. RESULTS: Experimental results conducted on 3,562 ultrasound images show that our new model greatly increases the accuracy and speed of the detection compared with the baseline model. The best mAP is 92.64%, and the fastest detection speed is 45.1 frames per second. CONCLUSIONS: This study proposed an effective method to diagnosis thyroid nodules automatically, which can meet the real-time requirements, indicating that its effectiveness and feasibility for future clinical application.

Ruthenium(II) polypyridyl complexes as dual inhibitors of telomerase and topoisomerase.

One novel ruthenium polypyridyl complex, [Ru(bpy)2(icip)](2+) (1), and two previously reported ruthenium polypyridyl complexes, [Ru(bpy)2(pdppz)](2+) ()2 and [Ru(bpy)2(tactp)](2+) (3) (bpy = 2,2'-bipyridine, icip = 2-(indeno[2,1-b]chromen-6-yl)-1H-imidazo[4,5-f][1,10]phenanthroline, pdppz = phenanthro[4,5-abc]dipyrido[3,2-h:2',3'-j]phenazine, tactp = 4,5,9,18-tetraazachryseno[9,10-b]-triphenylene), have been synthesised. As expected, these complexes show inhibition towards telomerase by inducing and stabilising the G-quadruplex structure, and behave as topoisomerase I/II poisons at the same time. Additionally, the acute and chronic cytotoxicities of the complexes are considered. Furthermore, cell apoptosis experiments are used to briefly study the mechanism. Because studies involving multi-target inhibition towards topoisomerase and telomerase of Ru(II) complexes have not been reported previously, the present research may help to develop innovative chemical strategies and therapies.

Novel ruthenium(II) polypyridyl complexes as G-quadruplex stabilisers and telomerase inhibitors.

Two novel ruthenium polypyridyl complexes, Ru[(bpy)2(pedppz)](2+) (1) and Ru[(bpy)2(pemitatp)](2+) (2) (bpy = 2'2-bipyridine, pdeppz = 10-(2-(piperidin-1-yl)ethoxy)dipyrido[3,2-a:2',3'-c]phenazine, pemitatp = 5-methoxy-1-(2-(piperidin-1-yl)ethyl)-isatino[1,2-b]-1,4,8,9-tetraazatriphenylene), bearing large planar π-delocalized aromatic systems with flexible chains have been synthesised and characterised. As expected, these complexes show inhibition towards telomerase by inducing and stabilising the G-quadruplex structure.

Visual specific luminescent probing of hybrid G-quadruplex DNA by a ruthenium polypyridyl complex.

A novel ruthenium(II) complex with specific luminescent selectivity towards hybrid G-quadruplex DNA was developed that can easily be distinguished by the naked eye without further treatment.