GLIDR promotes the progression of glioma by regulating the miR-4677-3p/MAGI2 axis.

Gliomas are the most common and fatal primary brain tumors. Growing evidence suggests that long non-coding RNAs (lncRNAs) constitute novel and potential therapeutic targets for glioma. However, the biological role of glioblastoma down-regulated RNA (GLIDR) in glioma remains largely elusive. In the current study, we used quantitative real-time polymerase chain reaction (qRT-PCR) to detect GLIDR expression in glioma cells. Cell counting kit 8 (CCK-8) assay, colony formation assay, JC-1 staining, and flow cytometry were used to evaluate the role of GLIDR in proliferation and apoptosis of glioma cells. Western blotting was performed to assess the effect of GLIDR on the level of apoptosis-related proteins. In addition, bioinformatics prediction, RNA immunoprecipitation (RIP), RNA pull-down, and luciferase reporter gene assays were used to study the regulatory mechanisms of GLIDR in glioma. GLIDR was found to be highly expressed in glioma cells and silencing of GLIDR inhibited cell proliferation and promoted apoptosis. Functionally, GLIDR bound to miR-4677-3p that directly targeted membrane-associated guanylate kinase, WW, and PDZ domain-containing protein 2 (MAGI2). Our data showed that GLIDR affects the proliferation and apoptosis of glioma cells by targeting miR-4677-3p to regulate the expression of MAGI2. In conclusion, our study determined the oncogenic role of GLIDR in glioma, which may provide a new perspective for the treatment of glioma.

DEK proto-oncogene is highly expressed in astrocytic tumors and regulates glioblastoma cell proliferation and apoptosis.

Astrocytic tumors are the most common neuroepithelial neoplasms with high relapse rate after surgery. Understanding the molecular mechanisms for astrocytic tumorigenesis and progression will lead to early diagnosis and effective treatment of astrocytic tumors. The DEK mRNA and protein expression in normal brain tissues and astrocytic tumors was quantified. To investigate DEK functions in tumor cells, DEK gene was silenced with siRNA in U251 glioblastoma cells. Cell proliferation, cell cycle and apoptosis were then measured. The expression and activity of key genes that regulate cell proliferation and apoptosis were also measured. We identified DEK as a high expressed gene in astrocytic tumor tissues. DEK expression level was positively correlated with the pathological grade of astrocytic tumors. Gene silencing of DEK in U251 glioblastomas inhibited cell proliferation and blocked cells at G0/G1 phase of cell cycle. DEK depletion also induced cell apoptosis, with up-regulated expression of P53 and P21 and down-regulated expression of Bcl-2 and C-myc. The Caspase-3 activity in U251 cells was also significantly increased after knockdown. Our results provided evidences that DEK regulates proliferation and apoptosis of glioblastomas. DEK gene silencing may induce apoptosis through P53-dependent pathway. Our data indicated DEK plays multiple roles to facilitate tumor growth and maintenance. It can be used as a potential target for astrocytic tumor diagnosis and gene therapy.

CRM197 in Combination With shRNA Interference of VCAM-1 Displays Enhanced Inhibitory Effects on Human Glioblastoma Cells.

CRM197 is a naturally nontoxic diphtheria toxin mutant that binds and inhibits heparin-binding epidermal growth factor-like growth factor. CRM197 serves as carrier protein for vaccine and other therapeutic agents. CRM197 also inhibits the growth, migration, invasion, and induces apoptosis in various tumors. Vascular cell adhesion molecule-1 (VCAM-1) is an important cell surface adhesion molecule associated with malignancy of gliomas. In this work, we aimed to investigate the role and mechanism of CRM197 combined with shRNA interference of VCAM-1 (shRNA-VCAM-1) on the migration, invasion, and apoptosis of glioblastoma cells. U87 and U251 human glioblastoma cells were treated with CRM197 (10 µg/ml) and shRNA interfering technology was employed to silence VCAM-1 expression. Cell viability, migration, invasiveness, and apoptosis were assessed with CCK8, Transwell and Annexin V-PE/7-AAD staining. Activation of cleaved caspase-3, 8, and 9, activity of matrix metalloproteinase-2/9 (MMP-2/9), and expression of phosphorylated Akt (p-Akt) were also checked. Results showed that CRM197 and shRNA-VCAM-1 not only significantly inhibited the cell proliferation, migration, invasion, but also promoted the apoptosis of U87 and U251 cells. Combined treatment of both displayed enhanced inhibitory effects on the malignant biological behavior of glioma cells. The activation of cleaved caspase-3, 8, 9 was promoted, activity of MMP-2 and MMP-9 and expression of p-Akt were inhibited significantly by the treatment of CRM197 and shRNA-VCAM-1 alone or in combination, indicating that the combination of CRM197 with shRNA-VCAM-1 additively inhibited the malignant behavior of human glioblastoma cells via activating caspase-3, 8, 9 as well as inhibiting MMP-2, MMP-9, and Akt pathway.

A 41-Year-Old Woman with a Late Cerebral Metastasis 16 Years After an Initial Diagnosis of Cutaneous Melanoma.

BACKGROUND Late cerebral metastasis more than 10 years after the diagnosis of cutaneous melanoma is very rare. This report is of a woman with late cerebral metastasis 16 years after an initial diagnosis of cutaneous melanoma. CASE REPORT A 41-year-old woman had been diagnosed with malignant melanoma 16 years prior from a biopsy of a dish-pattern tumor on the back, for which she received chemotherapy for 5 times (therapeutic regimen and medications were not available). She had not had a diagnosis of skin melanoma in the past 16 years. Before presentation to the Emergency Department, she had a progressive disturbance of consciousness for 6 weeks and sudden coma for 6 h. A head computed tomography scan indicated intracranial masses located at the right frontal and temporal lobes. The patient underwent surgery for tumor and hematoma removal. During surgery, dural metastasis with widespread dissemination in adjacent temporal bone, temporalis, and hypodermis was confirmed. Postoperative histopathology analysis confirmed the diagnosis of malignant melanoma metastasis. On the second day after surgery, the patient developed recurrent bleeding in the right frontal lobe, which led to deteriorated consciousness. She received hematoma evacuation and craniectomy and lived in a poor condition with drowsiness and hemiplegia of the left limb for 3 months and died 5 months after craniectomy. CONCLUSIONS This report has presented a rare occurrence of late cerebral metastasis 16 years after the initial diagnosis of a primary cutaneous melanoma. More recent primary melanoma of the skin was not identified, which supports the need for long-term follow-up of patients with a history of primary cutaneous melanoma.

KLF11 Expression Predicts Poor Prognosis in Glioma Patients.

PURPOSE: Glioma is a primary intracranial malignant tumor with high recurrence and mortality rates. It is very important to study the prognostic factors. KLF11 can function as an oncogene or a tumor suppressor, depending on the tumor and tissue types and the cancer stage. In this study, we aimed to determine whether KLF11 expression is related to the overall survival of glioma patients. PATIENTS AND METHODS: We investigated KLF11 expression in 116 glioma patients with different grades using Western blot and immunohistochemistry assay. We analyzed the patients with different glioma grades and KLF11 expression levels by Kaplan-Meier survival curves. Independent prognostic factors for poor overall survival were identified by univariate and multivariate analyses. RESULTS: There were 37 patients in KLF11 low expression group and 79 patients in high expression group. There was no difference in gender, age, tumor diameter or tumor location between two groups. The patients in KLF11 high expression group had higher ECOG score (P =0.025) and higher WHO grades (P =0.029). Western blot and immunohistochemistry assay showed KLF11 expression was significantly upregulated in glioma groups compared with normal brain tissues group (P < 0.05), and the expression in grades III-IV was significantly higher than those in grades I-II (P < 0.05). Kaplan-Meier survival curve analysis showed high KLF11 expression tended to reduce the overall survival (P < 0.05). After univariate and multivariate analyses, KLF11 expression (P =0.003) and age (P =0.007) were independent prognostic factors for poor survival in glioma patients. CONCLUSION: KLF11 expression was increased in glioma tissues, and high KLF11 expression was associated with poor prognosis. KLF11 expression was an independent prognostic factor for poor survival in glioma patients. KLF11 may serve as a novel prognostic marker for gliomas and as a novel treatment target.

Immune Infiltrates of m6A RNA Methylation-Related lncRNAs and Identification of PD-L1 in Patients With Primary Head and Neck Squamous Cell Carcinoma.

Background: The purpose of this study was to determine the association between m6A-modified lncRNAs, immune infiltration, and PD-L1 expression in patients with primary head and neck squamous cell carcinoma (HNSCC) and the prognostic value of m6A RNA methylation-related lncRNAs in HNSCC. Methods: We downloaded the RNA-seq transcriptome data and the clinical information for HNSCC from the TCGA databases and used consensus clustering analysis to divide the samples into two groups. To identify a risk signature, least absolute shrinkage and selection operator (LASSO) analyses were conducted. the association between m6A-modified lncRNAs, immune infiltration, and PD-L1 expression were detected by using the R packages. What is more, we used cBioPortal tools to identify genomic alterations and PD-L1 mutations and Gene set enrichment analysis (GSEA) was utilized to predict downstream access of two clusters. Results: Notably, lncRNAs play significant roles in tumorigenesis and development. In total, we identified two subtypes of HNSCC according to consensus clustering of the m6A RNA methylation-related lncRNAs, and the T, grade and age were proven to be related to the subtypes. The Cox regression and LASSO analyses identified a risk signature including GRHL3-AS1, AL121845.4, AC116914.2, AL513190.1. The prognostic value of the risk signature was then proven. The selected gene PD-L1 mutations and the immune infiltration in both groups were further explored. Conclusion: Collectively, our study elucidated the important role of m6A RNA methylation- related lncRNAs in tumor microenvironment of HNSCC. The proposed m6A RNA methylation- related lncRNAs might serve as crucial mediators of tumor microenvironment of HNSCC, representing promising therapeutic targets in improving immunotherapeutic efficacy.

Drinking habits and water sources with the incidence of cognitive impairment in Chinese elderly population: The Chinese Longitudinal Healthy Longevity Survey.

BACKGROUND: Little was known on whether drinking habits and water sources affected cognitive function. This study aimed to examine the associations of drinking habits and water sources with the incidence of cognitive impairment in the Chinese elderly population. METHODS: Data were drawn from the Chinese Longitudinal Healthy Longevity Survey. All participants aged ≥ 60 years at baseline were potential eligible. Cognitive function was measured using the Chinese version of the Mini-Mental State Examination (CMMSE). Participants with the CMMSE score ≤ 24 were identified as cognitive impairment. Drinking habits included preferring to drink boiled water or un-boiled water. Water sources included well, surface water, spring, and tap water. RESULTS: This study included 18034 participants. Participants drinking un- boiled water were more likely to develop cognitive impairment than those drinking boiled water (P< 0.001; HR: 1.269; and 95% CI: 1.128-1.427). Compared to drinking from a well, drinking from tap water at childhood, around aged 60 years, and present was associated with a lower incidence of cognitive impairment (all P< 0.001; HR: 0.672, 0.735, and 0.765; and 95% CI: 0.540-0.836, 0.686-0.788, and 0.723-0.810, respectively). LIMITATIONS: The underline mechanisms behind the associations of drinking habits and water sources with the incidence of cognitive impairment were not fully explained. CONCLUSIONS: Participants drinking un- boiled water were more likely to develop cognitive impairment. Meanwhile, compared to drinking from a well, drinking from tap water was associated with a lower incidence of cognitive impairment.

Ventriculoperitoneal shunt malfunction due to chronic cholecystitis: A case report.

RATIONALE: Ventriculoperitoneal shunt (VPS) is the most common treatment for idiopathic normal pressure hydrocephalus, a subtype of hydrocephalus characterized by gait disturbance, dementia, and urinary incontinence. However, while the malfunction of VPS is reported at a high rate, the involvement of chronic cholecystitis in shunt malfunction is rare. PATIENT CONCERNS: A 73-year-old woman with idiopathic normal pressure hydrocephalus who received a VPS but subsequently developed chronic cholecystitis. The patient suffered from drowsiness and was unable to walk. Her family found that she presented with poor appetite and was bloated. DIAGNOSES: Chronic cholecystitis was confirmed through abdominal computed tomography, which showed a swollen, and enlarged gallbladder, and flatulence. A head computed tomography scan indicated hydrocephalus with enlarged ventricular system and paraventricular edema. INTERVENTIONS: Laparoscopic cholecystectomy was performed successfully, requiring no further shunt manipulation. OUTCOMES: The patient's memory and cognitive ability were slightly impaired without a positive sign in the abdomen. No catheter or abdominal infection signs were observed during the following 3 months of follow-up. CONCLUSION: To the best of our knowledge, this report is the first to reveal that shunt malfunction may result from chronic cholecystitis, which induced the presently observed intra-abdominal hypertension.

Associations of health behaviors, food preferences, and obesity patterns with the incidence of mild cognitive impairment in the middle-aged and elderly population: An 18-year cohort study.

BACKGROUND: There were few studies to examine the associations of food preferences and obesity patterns with the incident mild cognitive impairment (MCI). Therefore, this study aimed to examine the associations of health behaviors, food preferences, and with the incidence of MCI. METHODS: All participants aged ≥ 55 years were potential eligible. The types of health behaviors and food preferences were recorded using the valid questionnaire. The obesity patterns were defined as follows:G-/A-, G+/A- or G-/A+, and G+/A+. The cognition tests included immediate and delayed recall, counting backward from 20, and serial 7 subtraction. The total cognitive score ranged from 0 to 27. Subjects with a score < 7 were considered as MCI. RESULTS: There were 8236 subjects included in this study. Martial arts, ping pong, and reading or writing were associated with the lower incident MCI (P = 0.039, 0.006, and 0.016, respectively). However, TV or computer usage was associated with the higher incident MCI (P = 0.029; HR: 1.455; and HR 95% CI: 1.040- 2.036). Fast foods, soft/sugared drinks, and salty snack foods increased the incident MCI (P< 0.001, = 0.032, and 0.002, respectively). G+/A- or G-/A+ and G+/A+ were associated with the lower incident MCI (P = 0.018 and < 0.001, respectively). LIMITATIONS: The basic mechanisms of health behaviors, food preferences, and obesity patterns on the risk of MCI were not fully explained. CONCLUSION: Reading or writing and G+/A+ were associated with the lower incident MCI. However, TV or computer usage and fast foods were associated with the higher incident MCI.

Advances and Prospects of Vasculogenic Mimicry in Glioma: A Potential New Therapeutic Target?

Vasculogenic mimicry (VM) is the formation of a "vessel-like" structure without endothelial cells. VM exists in vascular-dependent solid tumors and is a special blood supply source involved in the highly invasive tumor progression. VM is observed in a variety of human malignant tumors and is closely related to tumor proliferation, invasion, and recurrence. Here, we review the mechanism, related signaling pathways, and molecular regulation of VM in glioma and discuss current research problems and the potential future applications of VM in glioma treatment. This review may provide a new viewpoint for glioma therapy.

SUMOylation of PUM2 promotes the vasculogenic mimicry of glioma cells via regulating CEBPD.

Glioma is the most common form of primary central nervous malignant tumors. Vasculogenic mimicry (VM) is a blood supply channel that is different from endothelial blood vessels in glioma. VM is related to tumor invasion and metastasis. Therefore, it plays an important role to target therapy for glioma VM. Our experimental results showed abnormal expression of UBE2I, PUM2, CEBPD, and DSG2 in glioma cells. The Co-IP and Immunofluorescence staining were used to detect that PUM2 can be modified by SUMO2/3. The interaction between PUM2 and CEBPD mRNA was detected by the RIP assays. The interaction between transcription factor CEBPD and promoter region of DSG2 was detected by the ChIP assays and luciferase assays. The capacity for migration in glioma cells was observed by the laser holographic microscope. The capacity for invasion in glioma cells was detected by Transwell method. The VM in glioma cells was detected by three-dimensional cell culture method. The experimental results found that the upregulation of UBE2I in glioma tissues and cells promotes the SUMOylation of PUM2, which decreases not only the stability of PUM2 protein but also decreases the inhibitory effect of PUM2 on CEBPD mRNA. The upregulation of CEBPD promotes the binding to the upstream promoter region of DSG2 gene, further upregulates the expression of DSG2, and finally promotes the development of glioma VM. In conclusion, this study found that the UBE2I/PUM2/CEBPD/DSG2 played crucial roles in regulating glioma VM. It also provides potential targets and alternative strategies for combined treatment of glioma.

Clinical Presentation and Pathologic Characteristics of Pituitary Metastasis from Breast Carcinoma: Cases and a Systematic Review of the Literature.

BACKGROUND: It is rare for breast carcinoma to metastasize to the pituitary gland; this finding indicates extensive metastasis of the primary tumor. CASE DESCRIPTION: Herein, we present a 57-year-old patient with pituitary gland metastasis from breast cancer that was treated with extensive radical mastectomy 16 years prior. The pituitary was the sole site of metastasis. The patient was admitted with the chief complaint of blurred vision for 1 year and episodic headaches for 1 month. Magnetic resonance imaging revealed a solid mass in the sellar region with heterogenous contrast enhancement. The preoperative diagnosis was a pituitary adenoma. Neuroendoscopy-assisted tumor resection was conducted through a single-nostril sphenoid sinus approach. A pinkish-white, firm neoplasm was found, with an abundant blood supply and an indistinct boundary between the neoplasm and normal pituitary tissue; complete resection was achieved. The results of immunohistochemical analysis were positive for cytokeratin, Ki-67antigen, estrogen receptors, progesterone receptors, and prolactin-induced protein. The neoplasm was negative for spalt-like transcription factor 4, mammaglobin, and the alpha subunit of the glycoprotein hormones. These results were used to reach a final diagnosis of pituitary gland metastasis from a primary breast carcinoma. The patient's vision improved significantly after surgery, and no recurrence was detected during 1 year of follow-up. CONCLUSIONS: Pituitary gland metastasis is rare and difficult to differentiate from a pituitary adenoma without a pathologic diagnosis. Surgery is the first choice for treatment. Surgery, radiotherapy, and chemotherapy are combined with endocrine therapy to tailor treatment to the results of immunohistochemistry.

Protective effects of nigranoic acid on cerebral ischemia-reperfusion injury and its mechanism involving apoptotic signaling pathway.

The goal of this study was to assess the expression of poly ADP-ribose polymerase (PARP) and apoptosis-inducing factor (AIF) in the hippocampal CA1 region, and to find out whether nigranoic acid treatment exhibits protective effects on brain through PARP/AIF signaling pathway in cerebral ischemia-reperfusion animal model. Rats were randomly divided into three groups: Sham-surgery, ischemia-reperfusion, and nigranoic acid-treated. Rat models of middle cerebral artery occlusion were prepared using a way of thread occlusion. Rats in the nigranoic acid group were administered with 1 mg/kg intragastric nigranoic acid 6 and 2 h before brain ischemia, respectively. Following reperfusion, samples were collected at different time-points (6, 24, and 72 h) and each group was further divided into three subgroups. Apoptosis was measured using the terminal deoxynucleotidyl-transferase-mediated dUTP nick end labeling method. The protein expression levels of AIF and PARP were detected using Western blot and AIF mRNA quantity was evaluated using the reverse transcription-polymerase chain reaction. Apoptosis, levels of AIF and PARP protein expression, and levels of AIF mRNA expression were significantly increased in the ischemia-reperfusion group compared with the sham-surgery group. However, apoptosis and the expression levels of AIF protein, PARP protein, and AIF mRNA at different time-points were significantly decreased in the nigranoic acid-treated group compared with the model group. We can judge that nigranoic acid has a strong protective effect on rat cerebral ischemia-reperfusion injury, and acts by downregulating nerve cell apoptosis by preventing the overactivation of PARP and AIF nuclear translocation.

Suprasellar hemangioblastoma without von Hippel-Lindau disease: a case report and literature review.

Suprasellar hemangioblastoma (HBL) without von Hippel-Lindau (VHL) disease is extremely rare. A 51-year-old woman presented with headache and progressively deteriorating bilateral visual disturbance for 4 months. Magnetic resonance imaging (MRI) revealed a 2.5-cm solid mass in the suprasellar region with homogeneous contrast enhancement. Our preoperative presumptive diagnosis was meningioma. Resection of the tumor was achieved via a left pterional craniotomy. The tumor was reddish in appearance and relatively firm, and was extremely vascularized, which might provide extensive blood supply through small branches of the internal carotid artery. There was a clear border between the tumor and the pituitary stalk and optic nerves. Histopathologic examination showed that the tumor was well vascularized, consisting of a reticular mesh of numerous thin-walled capillaries and abundant stromal cells. Immunohistochemistry demonstrated the positive staining for CD34, vimentin (VIM), and neuron specific enolase (NSE) in the intratumoral capillaries, while negative staining of epithelial membrane antigen (EMA) and glial fibrillary acidic protein (GFAP) was observed. Based on these results, the patient was diagnosed as HBL. After the resection, the visual field defect in the left eye was markedly improved, and no tumor recurrence was noted in 1 year follow-up. When solid lesions are highly vascularized in the suprasellar region of patients, even though no VHL disease is present, the possibility of HBL should be taken into consideration. Moreover, craniotomy is a better treatment option for suprasellar HBL without VHL disease.

miR-101 acts as a tumor suppressor by targeting Kruppel-like factor 6 in glioblastoma stem cells.

BACKGROUND AND AIMS: Great interest persists in useful therapeutic targets in glioblastoma (GBM). Deregulation of microRNAs (miRNAs) expression has been associated with cancer formation through alterations in gene targets. In this study, we reported the role of miR-101 in human glioblastoma stem cells (GSCs) and the potential mechanisms. METHODS AND RESULTS: Quantitative real-time PCR showed that miR-101 expression was decreased in GSCs. Overexpression of miR-101 reduced the proliferation, migration, invasion, and promoted apoptosis of GSCs. One direct target of miR-101, the transcription factor Kruppel-like factor 6 (KLF6), was identified using the Dual-Luciferase Reporter Assay System, which mediated the tumor suppressor activity of miR-101. This process was coincided with the reduced expression of Chitinase-3-like protein 1 (CHI3L1) whose promoter could be bound with and be promoted by KLF6 demonstrated by luciferase assays and chromatin immunoprecipitation assays. The downregulation of CHI3L1 led to the inactivation of MEK1/2 and PI3K signal pathways. Furthermore, nude mice carrying the tumors of overexpressed miR-101 combined with knockdown of KLF6 produced the smallest tumors and showed the highest survival rate. CONCLUSIONS: Our findings provided a comprehensive analysis of miR-101 and further defining it as a potential therapeutic candidate for GBM.

Silencing of the DEK gene induces apoptosis and senescence in CaSki cervical carcinoma cells via the up-regulation of NF-κB p65.

The human DEK proto-oncogene has been found to play an important role in autoimmune disease, viral infection and human carcinogenesis. Although it is transcriptionally up-regulated in cervical cancer, its intracellular function and regulation is still unexplored. In the present study, DEK and IκBα [inhibitor of NF-κB (nuclear factor κB) α] shRNAs (short hairpin RNAs) were constructed and transfected into CaSki cells using Lipofectamine™. The stable cell line CaSki-DEK was obtained after G418 selection. CaSki-IκB cells were observed at 48 h after psiRNA-IκB transfection. The inhibitory efficiency of shRNAs were detected by RT (reverse transcription)-PCR and Western blot analysis. The proliferation activity of cells were measured using an MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide] assay, cell apoptosis was measured using an Annexin V/PI (propidium iodide) kit, the cell cycle was analysed by flow cytometry and cell senescence was detected using senescence ß-galactosidase staining. The intracellular expression of NF-κB p65 protein was studied by cytochemistry. The expression levels of NF-κB p65, p50, c-Rel, IκBα and phospho-IκBα protein were analysed by immunoblotting in whole-cell lysates, cytosolic fractions and nuclear extracts. The protein expression and activity of p38 and JNK (c-Jun N-terminal kinase) were also assayed. In addition, the NF-κB p65 DNA-binding activity was measured by ELISA. Following the silencing of DEK and IκBα, cell proliferation was inhibited, apoptosis was increased, the cell cycle was blocked in the G0/G1-phase with a corresponding decrease in the G2/M-phase, and cell senescence was induced. All of these effects may be related to the up-regulation of NF-κB p65 expression and its nuclear translocation.