The Effect of High CDC6 Levels on Predicting Poor Prognosis in Colorectal Cancer

<b><i>Introduction:</i></b> We investigated the function of cell division cycle 6 (CDC6) on the prognosis in colorectal carcinoma (CRC). <b><i>Methods:</i></b> CDC6 protein expression levels in 121 patients with colorectal cancer and adjacent normal mucosa were detected by immunohistochemistry. <b><i>Results:</i></b> Compared to adjacent normal tissues, CDC6 mRNA level was overexpressed in CRC tissues. Moreover, CDC6 protein levels were expressed up to 93.39% (113/121) in CRC tissues in the cell nucleus or cytoplasm. However, there were only 5.79% (7/121) in normal mucosal tissues with nuclear expression. CDC6 expression was significantly correlated with TNM stage and tumor metastasis. The 5-year survival rate was lower in the high CDC6 expression group than the low group. After silencing of CDC6 expression in SW620 cells, cell proliferation was slowed, the tumor clones were decreased, and the cell cycle was arrested in G1 phase. In multivariate analysis, increased CDC6 protein expression levels in colon cancer tissues were associated with cancer metastasis, TNM stage, and patient survival time. <b><i>Conclusion:</i></b> CDC6 is highly expressed in CRC, and downregulation of CDC6 can slow the growth of CRC cells in vitro. It is also an independent predictor for poor prognosis and may be a useful biomarker for targeted therapy and prognostic evaluation.

Colonic Mucosa Barrier Defects in Collagenous and Ischemic Colitis

Aims The subepithelial myofibroblasts (SEMFs) and the subepithelial band of macrophages (SEBM) are major components of the colonic mucosa barrier. Although their role in homeostasis is widely recognized, their contribution to disease states is largely unknown. The aim of the study was to explore histological characteristics of SEMFs and SEBM in collagenous and ischemic colitis.Methods Ten colonic biopsies of collagenous colitis, 10 of ischemic colitis and 10 control biopsies of normal mucosa were examined. SEMFs, SEBM and lamina propria macrophages were identified immunohistochemically by aSMA and CD68 respectively.ResultsIn collagenous colitis, SEMFs were rarely detectable in the collagenous band while in the lower lamina propria cell processes were formed. SEBM was preserved in areas with a collagenous layer up to 20μm. In thicker layers, it was fragmented and gradually disappeared in parallel with engulfment of enlarged macrophages. In the lower lamina propria macrophages were usually increased.In ischemic colitis, rounding, disintegration and extinction of SEMFs constituted successive alterations coinciding with crypt shrinkage and denudation. SEBM displayed total or almost total abolishment in areas with crypt damage and stroma fibrosis but also in sights with minimal changes.ConclusionIn collagenous colitis, alterations of mucosa barrier are related to collagenous layer thickness. SEMFs changes probably reflect derangement of differentiation and migration while SEMB alterations seem to be compensated by macrophage activation and numerical increase in lamina propria. The striking damage of mucosa barrier in ischemic colitis is indicative of its high sensitivity to hypoxia and hypoperfusion. The histological differences between collagenous colitis and ischemic colitis may be proven of differential diagnostic significance.

Deep Learning and Device-Assisted Enteroscopy: Automatic Detection of Gastrointestinal Angioectasia

Background and Objectives: Device-assisted enteroscopy (DAE) allows deep exploration of the small bowel and combines diagnostic and therapeutic capacities. Suspected mid-gastrointestinal bleeding is the most frequent indication for DAE, and vascular lesions, particularly angioectasia, are the most common etiology. Nevertheless, the diagnostic yield of DAE for the detection of these lesions is suboptimal. Deep learning algorithms have shown great potential for automatic detection of lesions in endoscopy. We aimed to develop an artificial intelligence (AI) model for the automatic detection of angioectasia DAE images. Materials and Methods: A convolutional neural network (CNN) was developed using DAE images. Each frame was labeled as normal/mucosa or angioectasia. The image dataset was split for the constitution of training and validation datasets. The latter was used for assessing the performance of the CNN. Results: A total of 72 DAE exams were included, and 6740 images were extracted (5345 of normal mucosa and 1395 of angioectasia). The model had a sensitivity of 88.5%, a specificity of 97.1% and an AUC of 0.988. The image processing speed was 6.4 ms/frame. Conclusions: The application of AI to DAE may have a significant impact on the management of patients with suspected mid-gastrointestinal bleeding.

ANALYSIS OF EXPRESSION OF MIRNA AND MRNA IN THE CELLULAR MATERIAL OF THE STOMACH LINING OBTAINED BY ESOPHAGOGASTRODUODENOSCOPY IN ORDER TO DETECT DYSPLASIA AND STOMACH CANCER

miRNA and mRNA are highly specific molecular markers, with their own unique expression profile for each type of tissue. They can become a valuable tool in clinical practice in addition to routine esophagogastroduodenoscopy.Purpose. To study the prospects of using a classifier based on miRNA and mRNA profiling in cytological samples for detecting dysplasia and stomach cancer, as well as to compare the obtained data with the results of profiling of 7 miRNAs used for the analysis of both histological and cytological material.Materials and methods. The study included 221 cytological preparations: 108 samples of adenocarcinoma, 27 samples of dysplasia, 86 samples of normal mucosa. The expression level of miRNA-145-5p, -150-5p, -21,-20a, -31-5p, -34a-5p, -375, -125b, -196b, -106b and mRNA of the following genes: TERT, CDKN2A, CKS2, FN1, was determined using real-time reverse transcription polymerase chain reaction. Quantitative comparison of two independent samples for was performed using the Mann-Whitney test. The samples were stratified into different groups using the C5.0 algorithm.Results: when examining cytological samples in cases of miRNA-125b, 145, -196b, -21, -375 and TERT, СKS2, FN1 mRNA, there was achieved a high level of significance of differences in the cancer/norm group; miRNA-375 and FN1 mRNA in the cancer/dysplasia group and miRNA-145, -196b, -20a, and СKS2, TERT mRNA in the norm/dysplasia groups. When comparing the results for the 7 miRNAs that were used to analyze both histological and cytological material, several differences was noted.Conclusion. The practical use of miRNA and mRNA expression (in cytological material data mining technology does not allow differentiating dysplasia and cancer but can help in identifying "high-risk patients" who should repeat esophagogastroduodenoscopy with multifocal forceps biopsy, with mandatory molecular genetic research.

A Gene-Based Machine Learning Classifier Associated to the Colorectal Adenoma—Carcinoma Sequence

Colorectal cancer (CRC) carcinogenesis is generally the result of the sequential mutation and deletion of various genes; this is known as the normal mucosa–adenoma–carcinoma sequence. The aim of this study was to develop a predictor-classifier during the “adenoma-carcinoma” sequence using microarray gene expression profiles of primary CRC, adenoma, and normal colon epithelial tissues. Four gene expression profiles from the Gene Expression Omnibus database, containing 465 samples (105 normal, 155 adenoma, and 205 CRC), were preprocessed to identify differentially expressed genes (DEGs) between adenoma tissue and primary CRC. The feature selection procedure, using the sequential Boruta algorithm and Stepwise Regression, determined 56 highly important genes. K-Means methods showed that, using the selected 56 DEGs, the three groups were clearly separate. The classification was performed with machine learning algorithms such as Linear Model (LM), Random Forest (RF), k-Nearest Neighbors (k-NN), and Artificial Neural Network (ANN). The best classification method in terms of accuracy (88.06 ± 0.70) and AUC (92.04 ± 0.47) was k-NN. To confirm the relevance of the predictive models, we applied the four models on a validation cohort: the k-NN model remained the best model in terms of performance, with 91.11% accuracy. Among the 56 DEGs, we identified 17 genes with an ascending or descending trend through the normal mucosa–adenoma–carcinoma sequence. Moreover, using the survival information of the TCGA database, we selected six DEGs related to patient prognosis (SCARA5, PKIB, CWH43, TEX11, METTL7A, and VEGFA). The six-gene-based classifier described in the current study could be used as a potential biomarker for the early diagnosis of CRC.

The Expression of ephrinA1/ephA2 Receptor Increases in Chronic Rhinosinusitis and ephrinA1/ephA2 Signaling Affects Rhinovirus-Induced Innate Immunity in Human Sinonasal Epithelial Cells

EphA2 receptor and its ephrin ligands are involved in virus infection, epithelial permeability, and chemokine secretion. We hypothesized that ephrinA1/ephA2 signaling participates in rhinovirus (RV)-induced antiviral immune response in sinonasal mucosa of patients with chronic rhinosinusitis (CRS). Therefore, we investigated the expression of ephrinA1/ephA2 in normal and inflamed sinonasal mucosa and evaluated whether they regulate chemokine secretion and the production of antiviral immune mediators including interferons (IFNs) in RV-infected human primary sinonasal epithelial cells. For this purpose, the expression and distribution of ephrinA1/ephA2 in sinonasal mucosa were evaluated with RT-qPCR, immunofluorescence, and western blot. Their roles in chemokine secretion and the production of antiviral immune mediators such as type I and III IFNs, and interferon stimulated genes were evaluated by stimulating ephA2 with ephrinA1 and inactivating ephA2 with ephA2 siRNA or inhibitor in cells exposed to RV and poly(I:C). We found that ephrinA1/ephA2 were expressed in normal mucosa and their levels increased in inflamed sinonasal mucosa of CRS patients. RV infection or poly(I:C) treatment induced chemokine secretion which were attenuated by blocking the action of ephA2 with ephA2 siRNA or inhibitor. The production of antiviral immune mediators enhanced by rhinovirus or poly (I:C) is increased by blocking ephA2 compared with that of cells stimulated by either rhinovirus or poly(I:C) alone. In addition, blocking ephA2 attenuated RV replication in cultured cells. Taken together, these results describe a novel role of ephrinA1/ephA2 signaling in antiviral innate immune response in sinonasal epithelium, suggesting their participation in RV-induced development and exacerbations of CRS.

Association between Endoscopic Findings Vs. Serology Findings of Patients with Suspected Celiac Disease

Objective: To determine the association between endoscopic findings vs. serology findings of patients with suspected celiac disease Methods: All the suspected cases (based on their clinical manifestations) of celiac disease were initially recruited having age >14 years and <40 years of both gender. Patients who did not willing to participate, patients already taking gluten diet for more than 3 months, patients with other causes of chronic diarrhea and alternate diagnosis like thyrotoxicosis, whipple’s disease, giardiasis, patients with drug induced diarrhea, patients in whom we cannot perform endoscopy, pregnant women, and patients already diagnosed cases of celiac disease were excluded from this study. Celiac disease was confirmed based on positive anti-tTG antibodies. Endoscopic evaluation of duodenum was performed in all positive cases. Results: A total of 50 patients were recruited for final analysis. Diagnostic accuracy of endoscopy was 34.6%. Young population (31.14±6.07 years) with females predominance (72%, n=36) were more common than males. The most common symptoms were presence of chronic diarrhea (74%, n=37) followed by abdominal pain (52%, n=26), nausea & vomiting (34%, n=17), and least common was presence of constipation (2%, n=1). On endoscopic evaluation, out of 50 positive anti-tTG antibodies cases, 24 had normal mucosa while partial villous atrophy observed in 15 (30%) cases and total villous atrophy observed in 11 cases (22%). Conclusions: Celiac disease was more prevalent in young females and patients usually presents with history of chronic diarrhea. Anti-tTG antibodies have more diagnostic value than duodenal endoscopy. Villous atrophy was found in more than 50% of the patients who were diagnosed with celiac disease.

Modulating Properties of Piroxicam, Meloxicam and Oxicam Analogues against Macrophage-Associated Chemokines in Colorectal Cancer

The mechanisms underlying the antineoplastic effects of oxicams have not been fully elucidated. We aimed to assess the effect of classic and novel oxicams on the expression/secretion of macrophage-associated chemokines (RTqPCR/Luminex xMAP) in colorectal adenocarcinoma cells, and on the expression of upstream the non-steroidal anti-inflammatory drug (NSAID)-activated genes NAG1, NFKBIA, MYD88, and RELA, as well as at the chemokine profiling in colorectal tumors. Meloxicam downregulated CCL4 9.9-fold, but otherwise the classic oxicams had a negligible/non-significant effect. Novel analogues with a thiazine ring substituted with arylpiperazine and benzoyl moieties significantly modulated chemokine expression to varying degree, upregulated NAG1 and NFKBIA, and downregulated MYD88. They inhibited CCL3 and CCL4, and their effect on CCL2 and CXCL2 depended on the dose and exposure. The propylene linker between thiazine and piperazine nitrogens and one arylpiperazine fluorine substituent characterized the most effective analogue. Only CCL19 and CXCL2 were not upregulated in tumors, nor was CXCL2 in tumor-adjacent tissue compared to normal mucosa. Compared to adjacent tissue, CCL4 and CXCL2 were upregulated, while CCL2, CCL8, and CCL19 were downregulated in tumors. Tumor CCL2 and CCL7 increased along with advancing T and CCL3, and CCL4 along with the N stage. The introduction of arylpiperazine and benzoyl moieties into the oxicam scaffold yields effective modulators of chemokine expression, which act by upregulating NAG1 and interfering with NF-κB signaling.

Lymphoma presented as dysphagia: a diagnosis hard to swallow

An 18-year-old woman presented with progressive oesophageal dysphagia, weight loss and night sweats over a 6-month period. Oesophagogastroduodenoscopy revealed a diffuse luminal narrowing with normal mucosa, whose biopsies were inconclusive. A cervical and thoracic CT scan showed a thickening of the upper oesophagus, densification of the mediastinal fat, several adenopathies and a 4.3×2.4 cm mass with infiltrative appearance and heterogeneous enhancement in right cervical paravertebral location. Positron emission tomography-CT showed marked increased fluorodeoxyglucose uptake in supradiaphragmatic lymph nodes, pleuropulmonary tissue, paraspinal musculature and bone marrow. Imaging-guided and surgical incisional biopsies of the paravertebral mass were inconclusive. During hospitalisation, she developed right cervicobrachial paraesthesia. Only excisional biopsy of the mass allowed the diagnosis of high-grade B-cell lymphoma not otherwise specified, Ann Arbor stage IV-B. The patient underwent chemotherapy with R-CHOP (rituximab, cyclophosphamide, doxorubicin, vincristine and prednisolone), followed by R-EPOCH (rituximab, etoposide, prednisone, vincristine, cyclophosphamide and doxorubicin hydrochloride). Follow-up at 12 months revealed complete response.