Pan-Cancer Analysis and Experimental Validation of CEND1 as a Prognostic and Immune Infiltration-Associated Biomarker for Gliomas.

Cell cycle exit and neuronal differentiation 1 (CEND1), highly expressed in the brain, is a specific transmembrane protein which plays a tumor suppressor role. This study is performed to investigate the role of CEND1 in various cancers through pan-cancer analysis, and further investigate its functions in gliomas by cell experiments. The expression and subcellular localization of CEND1 in different cancer types were analyzed utilizing the data from the GEPIA, UCSC, UALCAN and HPA databases. Relationships of CEND1 expression with prognosis, immunomodulation-related genes, immune checkpoint genes, microsatellite instability (MSI), tumor mutation burden (TMB) and RNA modifications were analyzed based on the TCGA database. The ESTIMATE algorithm was utilized to evaluate tumors' StromalScore, Immune Score, and ESTIMATES Score. The cBioPortal database was employed to analyze the categories and frequencies of CEND1 gene alterations. Biological functions and co-expression patterns of CEND1 in gliomas were explored using the LinkedOmics database, and Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were conducted. The interactions between CEND1 and drugs were explored employing the Comparative Toxicogenomics Database and molecular docking technology. Cell experiments were conducted to analyze triptonide's effects on glioma cells through CCK-8, flow cytometry and qRT-PCR. CEND1 was lowly expressed in gliomas, and high CEND1 expression was correlated to better overall survival of glioma patients (HR = 0.65, P = 0.02). Deep deletion was the main type of hereditary change of CEND1 mutation. CEND1 expression was markedly associated with immune infiltration, TMB, MSI, and RNA modification in various tumors (r > 0.3, P < 0.05). CEND1 co-expressed genes in gliomas were markedly correlated with immune responses and cell cycle (FDR < 0.05). Triptonide could bind well to CEND1 (-5.0 kcal/mol), and triptonide could facilitate CEND1 expression in glioma cells and cell apoptosis, and block the cell cycle progression (P < 0.05). CEND1 serves as a potential biomarker for pan-cancer. Particularly in gliomas, CEND1 is a key regulator of cell apoptosis and cell cycle, and a potential target for glioma treatment.

Lymphocytic interstitial pneumonia presenting with a ground glass nodule: A case report and literature review.

RATIONALE: Lymphocytic interstitial pneumonia (LIP) is a rare benign lymphoproliferative disorder, often associated with autoimmune diseases. Most LIPs present with multiple bronchial cysts and diffuse interstitial infiltration. It is histologically characterized by widespread diffuse lymphocytic infiltration of the pulmonary interstitium, and the enlargement and widening of the alveolar septum. PATIENT CONCERNS: A 49-year-old woman was admitted to hospital for finding pulmonary nodules for more than 2 months. 3D imaging chest computed tomography (CT) examination of both lungs showed that there was a middle lobe of the right lung with a size of about 1.5 cm × 1.1 cm ground-glass nodules. DIAGNOSES: A single operating port thoracoscopic wedge resection biopsy of a right middle lung nodule was performed. The pathology showed diffuse lymphocytic infiltration with varying numbers of small lymphocytes, plasma cells, macrophages and histiocytes infiltrating the alveolar septa, widened and enlarged alveolar septa, and scattered lymphoid follicles. Immunohistochemically, CD20 positive in follicular area, CD3 positive in interfollicular area. LIP was considered. INTERVENTIONS: The patient was regularly followed without any specific treatment. OUTCOMES: Follow-up chest CT showed no significant abnormalities in the lungs 6 months after surgery. LESSONS: To the best of our knowledge, our case may be the second reported case of a patient with LIP presenting with a ground glass nodule on chest CT, and it is speculated that the ground glass nodule may be an early manifestation of idiopathic LIP.

Circ_0001982 Up-regulates the Expression of E2F1 by Adsorbing miR-1205 to Facilitate the Progression of Glioma.

This study was aimed at probing into the regulatory effects of circular RNA (circRNA)_0001982 on glioma cell proliferation, migration, invasion, and cell cycle, and its underlying mechanism. CircRNA expression profile of glioma tissues/normal brain tissues was downloaded from the Gene Expression Omnibus (GEO) database and analyzed. Circ_0001982, microRNA (miRNA, miR)-1205, and E2F transcription factor 1 (E2F1) expressions in glioma tissues and cell lines were quantified using quantitative real-time polymerase chain reaction (qRT-PCR) and/or Western blot. Glioma cell proliferation, migration, invasion, and cell cycle were detected employing cell counting kit-8 (CCK-8), 5-Ethynyl-2'-deoxyuridine (EdU), scratch-healing, Transwell, and flow cytometry assays, respectively. The targeting relationships between miR-1205 and circ_0001982, and miR-1205 and E2F1 3'UTR were verified using bioinformatics, dual-luciferase reporter experiments, and RNA immunoprecipitation (RIP) assay. Pearson's correlation analysis was applied to detect the correlations among circ_0001982, miR-1205, and E2F1 expression levels. Circ_0001982 expression level was increased in glioma tissues and correlated with larger tumor size. Circ_0001982 overexpression enhanced glioma cell proliferation, migration, and invasion, and accelerated cell cycle progression while knocking down circ_0001982 exerted opposite effects. Circ_0001982 directly targeted miR-1205, and miR-1205 directly targeted E2F1. Besides, circ_0001982 could up-regulate E2F1 expression via repressing miR-1205 expression. Circ_0001982 accelerates glioma progression by modulating the miR-1205/E2F1 axis.

Up-regulation of miR-663a inhibits the cancer stem cell-like properties of glioma via repressing the KDM2A-mediated TGF-ß/SMAD signaling pathway.

Emerging reports have shown that microRNAs (miRNAs) function as vital regulators in tumor development via modulating gene expression at the posttranscriptional level. Here, we explored the role and underlying mechanism of miR-663a in the proliferation, migration, invasion, and cancer stem cell-like (CSC) properties of glioma cells. Quantitative reverse transcription PCR (qRT-PCR) was implemented to detect miR-663a expression in glioblastoma tissues and the adjacent normal tissues. Additionally, gain- and loss-of-function assays of miR-633a were performed on U-251 MG cells or human primary glioblastoma cancer cells (pGBMC1). Cell proliferation, migration, invasion, CSC properties, and profiles of stem cell markers (including CD133, CD44) were examined by the MTT assay, Transwell assay, tumorsphere experiment, and Western blotting, respectively. The dual-luciferase reporter gene assay was performed to testify the targeted relationship between miR-663a and lysine demethylase 2A (KDM2A). The results showed that miR-663a was down-regulated in glioblastoma tissues and cells. Overexpressing miR-663a repressed the proliferation, migration, invasion, CSC properties of U-251 MG cells and pGBMC1, while miR-663a knockdown had the opposite effects. The in-vivo experiment confirmed that miR-663a repressed the growth of U-251 MG cells in nude mice. When cocultured with THP1 cells, U-251 MG cells gained enhanced proliferation, migration, invasion, and CSC properties. MiR-633a overexpression reversed THP1-mediated effects on U-251 MG cells, and reduced the "M2" polarization of THP1 cells. What's more, Mechanistically, KDM2A was targeted by miR-663a. KDM2A knockdown suppressed the progression and CSC properties of U-251 MG cells in vitro, and dampened TGF-ß. Overall, those data revealed that up-regulating miR-663a reduced glioma progression by inhibiting the KDM2A-mediated TGF-ß/Smad pathway.

Hyalinizing trabecular tumor of the thyroid: a clinicopathological analysis of four cases and review of the literature.

OBJECTIVE: To explore the clinicopathological features, diagnosis and differential diagnosis of hyalinizing trabecular tumor (HTT) of the thyroid. METHODS: The four HTT specimens were collected including demographics, clinical information, relevant images, the extent of thyroidectomy, the follow-up and representative pathological data of tumors were available for analysis. In addition, the immumohistochemical staining related to the tumor as well as the BRAF and N-ras mutation analysis were analysed. RESULTS: The mean age of four patients was 47 years old and the mean size of the tumor was 2.8 cm. Most of the patients were asymptomatic, while detecting incidentally by using neck ultrasound test. Ultrasound imaging of all cases showed demarcated substantial hypoechoic nodules in ipsilateral thyroid lobe. Computed Tomography (CT) showed a clear low density shadow in the affected thyroid lobe. Tumors of three cases were located at the left, but the other one was located at the right thyroid gland with a complete fibrous capsule. The cytological features resembled papillary thyroid carcinoma (PTC). The histological test indicated that the tumors had characteristic of trabecular growth pattern with hyalinizing material. The tumor cells were in shape of polygonal, oval or high columnar with an acidophilic or clear cytoplasm. The nuclei were oval with inconspicuous small nucleoli, prominent grooves and pseudoinclusion body in cell nucleus. Mitosis and psammoma bodies were rare to be observed. Cytoplasmic "yellow bodies" were frequently observed. The hyaline material was prominent, with positive periodic acid-Schiff (PAS) and negative Congo red staining. Immunohistochemically, tumor cells were positive for thyroglobulin (Tg), thyroid transcription factor-1 (TTF-1), CD56 and negative for calcitonin, cytokeratin 19 (CK19), HBME-1, S-100 and synaptophysin (SyN). Chromogranin A (CgA) and galectin-3 were expressed weakly in some cases. Staining with the MIB-1 antibody showed membranous/cytoplasmic immunoreactivity. Whereas, another clone of Ki-67 (SP6) showed a common nuclear pattern with an index of <1%. None of the four cases exhibited the BRAF V600E protein reactivity. Gene mutation analysis demonstrated no BRAF and N-ras mutation. There was no evidence of local recurrence or metastasis after 6 to 36 months of follow-up. CONCLUSIONS: HTT is an uncommon thyroid tumor with very low malignant potential. It has no particular clinical features, so it's often misdiagnosed in fine needle aspiration cytology (FNAC)/Ultrasonography-guided fine needle aspiration cytology (US-FNAC) and frozen section (FS). Its final diagnosis mainly relies on typical histopathological features and characteristic expression pattern of MIB-1 immunohistochemical staining.

Co-expression of midkine and pleiotrophin predicts poor survival in human glioma.

The aim of this study was to investigate whether co-expression of midkine (MK) and pleiotrophin (PTN) has prognostic relevance in human gliomas. Immunohistochemistry was used to investigate the expression of MK and PTN proteins in 168 patients with gliomas. The levels of MK and PTN mRNA in glioma tissues and paratumor tissues were evaluated in 45 paired cases by quantitative real-time polymerase chain reaction (qRT-PCR). Kaplan-Meier survival analysis was performed to assess prognostic significance. The expression levels of MK and PTN proteins in glioma tissue were both significantly higher (both p<0.001) than those in paratumor tissues on immunohistochemistry analysis, which was confirmed by qRT-PCR analysis. Additionally, the overexpression of either MK or PTN was significantly associated with the World Health Organization Grade (p=0.001 and 0.034, respectively), low Karnofsky Performance Status (KPS) score (p=0.022 and 0.001, respectively), time to recurrence (p=0.043 and 0.011, respectively) and poor overall survival (p=0.018 and 0.001, respectively). Multivariate Cox proportional-hazards regression analysis revealed that increased expressions of MK and PTN were both independent prognostic factors for poor overall survival (p=0.030 and 0.022, respectively). Furthermore, the co-expression of MK and PTN was more significantly (p=0.003) associated with adverse prognosis in patients with gliomas than the respective expression of MK or PTN alone. To our knowledge, these findings are the first to indicate that the co-expression of MK and PTN is significantly correlated with prognosis in glioma patients, suggesting that the co-expression of these proteins may be used as both an early diagnostic and independent prognostic marker.