The Diagnostic Value of Serum miRNA-221-3p, miRNA-382-5p, and miRNA-4271 in Ischemic Stroke.

BACKGROUND: Circulating miRNAs have been demonstrated as biomarkers for a number of diseases. In the present study, we aimed to compare the serum levels of the miRNA-221-3p, miRNA-382-5p, and miRNA-4271 between ischemic stroke and healthy control subjects and explore the potential roles as noninvasive biomarkers in the diagnosis of ischemic stroke. METHODS: Seventy-eight patients with ischemic stroke (60 ± 10.47 years old) and 39 healthy control subjects (61 ± 5.14 years old), age and sex matched, were recruited into the present study. The circulating miRNA levels were determined by quantitative real-time polymerase chain reaction using miRNA qPCR Assay Kit (CW Biotech, Beijing, China). Receiver operating characteristic (ROC) curves were analyzed using the SPSS software package (version 17) (SPSS Inc., Chicago, IL). RESULTS: Circulating serum miRNA-221-3p and miRNA-382-5p levels were significantly lower in patients with ischemic stroke compared to healthy control subjects, whereas there was no significant difference in the serum levels of miRNA-4271 between the stroke patients and healthy controls (P > .05). ROC curves revealed the areas under the curve for circulating miRNA-221-3p, miRNA-382-5p, and miRNA-4271 to be .8106, .7483, and .6317 in ischemic stroke patients compared with healthy volunteers, respectively. There were no correlations between circulating miRNAs and laboratory determinations except that the levels of circulating miRNA-4271 were positively correlated with glucose (r = .274, P = .031). CONCLUSIONS: Our findings suggest that serum circulating miRNA-221-3p and miRNA-382-5p might be used as potential noninvasive biomarkers for the diagnosis of ischemic stroke.

[MiR-211 promotes invasion of hepatoma cells by targeting estrogen receptor alpha].

OBJECTIVE: To investigate the regulatory functions and molecular mechanisms of miR211 in hepatocellular carcinoma (HCC). METHODS: Real-time reverse transcription-PCR was used to analyze the expression of miR-211 in 20 paired clinical specimens of HCC and adjacent noncancerous tissues.QGY7703 and HepG2 cells with stimulation or inhibition of miR-211 expression were used to evaluate the effects on malignant phenotypes with the transwell invasion assay. Candidate target genes of miR-211 were identified by bioinformatic screening and verified by the EGFP report assay, real-time PCR and western blotting. Moreover, the regulatory functions of miR-211 on the target genes were investigated by RNA interference and cell phenotype assays. RESULTS: miR-211 was up-regulated in HCC tissue specimens (t =6.26, P < 0.01).HCC cells overexpressing miR-211 showed greater invasive capacity than cells with inhibited expression (QGY-7703:t =12.59, P < 0.01; HepG2: t =17.82, P < 0.01). Estrogen receptor a (ESR1) was identified and validated as a target gene of miR-211; knockdown of ESR 1 promoted HCC invasive capacity (QGY-7703:t =8.97, P < 0.01; HepG2:t =29.31, P < 0.01). CONCLUSION: miR-211 promotes invasion of carcinoma cells by directly targeting ESR1.

Identification of ferroptosis related biomarkers and immune infiltration in Parkinson's disease by integrated bioinformatic analysis.

BACKGROUND: Increasing evidence has indicated that ferroptosis engages in the progression of Parkinson's disease (PD). This study aimed to explore the role of ferroptosis-related genes (FRGs), immune infiltration and immune checkpoint genes (ICGs) in the pathogenesis and development of PD. METHODS: The microarray data of PD patients and healthy controls (HC) from the Gene Expression Omnibus (GEO) database was downloaded. Weighted gene co-expression network analysis (WGCNA) was processed to identify the significant modules related to PD in the GSE18838 dataset. Machine learning algorithms were used to screen the candidate biomarkers based on the intersect between WGCNA, FRGs and differentially expressed genes. Enrichment analysis of GSVA, GSEA, GO, KEGG, and immune infiltration, group comparison of ICGs were also performed. Next, candidate biomarkers were validated in clinical samples by ELISA and receiver operating characteristic curve (ROC) was used to assess diagnose ability. RESULTS: In this study, FRGs had correlations with ICGs, immune infiltration. Then, plasma levels of LPIN1 in PD was significantly lower than that in healthy controls, while the expression of TNFAIP3 was higher in PD in comparison with HC. ROC curves showed that the area under curve (AUC) of the LPIN1 and TNFAIP3 combination was 0.833 (95% CI: 0.750-0.916). Moreover, each biomarker alone could discriminate the PD from HC (LPIN1: AUC = 0.754, 95% CI: 0.659-0.849; TNFAIP3: AUC = 0.754, 95% CI: 0.660-0.849). For detection of early PD from HC, the model of combination maintained diagnostic accuracy with an AUC of 0.831 (95% CI: 0.734-0.927), LPIN1 also performed well in distinguishing the early PD from HC (AUC = 0.817, 95% CI: 0.717-0.917). However, the diagnostic efficacy was relatively poor in distinguishing the early from middle-advanced PD patients. CONCLUSION: The combination model composed of LPIN1 and TNFAIP3, and each biomarker may serve as an efficient tool for distinguishing PD from HC.

Identification and validation of key molecules associated with humoral immune modulation in Parkinson's disease based on bioinformatics.

Objective: Parkinson's disease (PD) is the most common neurodegenerative movement disorder and immune-mediated mechanism is considered to be crucial to pathogenesis. Here, we investigated the role of humoral immune regulatory molecules in the pathogenesis of PD. Methods: Firstly, we performed a series of bioinformatic analyses utilizing the expression profile of the peripheral blood mononuclear cell (PBMC) obtained from the GEO database (GSE100054, GSE49126, and GSE22491) to identify differentially expressed genes related to humoral immune regulatory mechanisms between PD and healthy controls. Subsequently, we verified the results using quantitative polymerase chain reaction (Q-PCR) and enzyme-linked immunosorbent assay (ELISA) in clinical blood specimen. Lastly, receiver operating characteristic (ROC) curve analysis was performed to determine the diagnostic effects of verified molecules. Results: We obtained 13 genes that were mainly associated with immune-related biological processes in PD using bioinformatic analysis. Then, we selected PPBP, PROS1, and LCN2 for further exploration. Fascinatingly, our experimental results don't always coincide with the expression profile. PROS1 and LCN2 plasma levels were significantly higher in PD patients compared to controls (p < 0.01 and p < 0.0001). However, the PPBP plasma level and expression in the PBMC of PD patients was significantly decreased compared to controls (p < 0.01 and p < 0.01). We found that PPBP, PROS1, and LCN2 had an area under the curve (AUC) of 0.663 (95%CI: 0.551-0.776), 0.674 (95%CI: 0.569-0.780), and 0.885 (95%CI: 0.814-0.955). Furthermore, in the biological process analysis of gene ontology (GO), the three molecules were all involved in humoral immune response (GO:0006959). Conclusions: In general, PPBP, PROS1, and LCN2 were identified and validated to be related to PD and PPBP, LCN2 may potentially be biomarkers or therapeutic targets for PD. Our findings also provide some new insights on the humoral immune modulation mechanisms in PD.

Exploring the potential value of miR-148b-3p, miR-151b and miR-27b-3p as biomarkers in acute ischemic stroke.

Cerebrovascular disease is the main cause of death in the world. Here, we explored whether circulating serum miR-148b-3p, miR-151b and miR-27b-3p could be as potential diagnostic biomarkers for diagnosing acute ischemic stroke. Seventy-seven IS patients and forty-two healthy controls matched for age and sex were enrolled in the present study. Blood samples were drawn from IS patients within the 24 h. The correlation analysis was performed by Spearman. The ability to distinguish patients from healthy controls was determined by receiver operating characteristic (ROC) curve. The expression of circulating serum miR-148b-3p was significantly decreased, whereas miR-151b and miR-27b-3p were elevated significantly compared with controls. ROC analysis showed area under the ROC curve (AUC) of miR-148b-3p, miR-151b and miR-27b-3p to be 0.6647, 0.6852 and 0.6657, respectively. While the AUC increased to 0.8103 for the combination of miR-148b-3p and miR-27b-3p. Blood miR-151b level was negatively correlated with insulin-like growth factor-1 (IGF-1), and miR-27b-3p level was negatively correlated with IGF-1 and insulin-like growth factor binding protein-3, respectively. Our findings suggest that miR-148b-3p, miR-151b and miR-27b-3p may serve as blood-based biomarkers for diagnosing ischemic stroke patients, and the combination of miR-148b-3p and miR-27b-3p may be more powerful.

Knockdown of BCL6 Inhibited Malignant Phenotype and Enhanced Sensitivity of Glioblastoma Cells to TMZ through AKT Pathway.

BACKGROUND: BCL6 was a critical prooncogene of human B-cell lymphomas which promoted tumor progress and contributed to malignant behavior in several kinds of cancers. This study was to detect the expression of BCL6 and its biological effect on glioma. METHODS: RT-PCR and Western blot were used to detect the expression of BCL6 mRNA and protein in tissues and glioblastoma cell lines. The expression of BCL6 was knockdown in two glioblastoma cell lines (U87 and U251) using BCL6 shRNA. The CCK8, colony-formation, flow cytometry, Transwell, and wound-healing assays were used to evaluate the malignant phenotypic change of glioblastoma cells. RESULTS: The expression of BCL6 was higher in glioma tissues and glioblastoma cell lines than normal tissues. Knockdown of BCL6 expression reduced the proliferation, migration, and invasion of glioblastoma cells. Moreover, knockdown of BCL6 changed expression of proteins related to malignant behaviors of glioblastoma cells. The suppression of BCL6 could increase chemosensitivity of U87 and U251 to temozolomide. Downregulation of BCL6 levels suppressed the expression of BCL2, cyclin D1, MMP2, and MMP9 proteins as well as two classic signaling pathway proteins p-AKT and p-ERK. Simultaneously, BAX and p21 protein levels were upregulated along with knockdown of BCL6. CONCLUSIONS: Our results indicated that BCL6 may be a tumor oncogene involved in the progression of glioma via affecting AKT and MAPK signaling pathways.

Astragaloside IV reversed the autophagy and oxidative stress induced by the intestinal microbiota of AIS in mice.

Acute ischaemic stroke (AIS) seriously affects patient quality of life. We explored the role of the intestinal microbiota on oxidative stress and autophagy in stroke, and Astragaloside IV (AS-IV) reversed the changes induced by intestinal microbiota. We determined the characteristics of the intestinal microbiota of AIS and transient ischaemic attack (TIA) patients by 16S sequencing and found that the structure and diversity of the intestinal microbiota in patients with AIS and TIA were significantly different from those in healthy subjects. Specifically, the abundance of genus Bifidobacterium, Megamonas, Blautia, Holdemanella, and Clostridium, content of homocysteine and triglyceride was increased significantly, thus it may be as a potential mechanism of AIS and TIA. Furthermore, germ-free mice were infused intracolonically with fecal supernatants of TIA and AIS with/without feed AS-IV for 12 weeks, and we found that the feces of AIS up-regulated the autophagy markers Beclin-1, light chain 3 (LC3)-II and autophagy-related gene (Atg)12, and the expression of reactive oxygen species (ROS) and NADPH oxidase 2/4 (NOX2/4), malondialdehyde (MDA), however, the expression of total antioxidant capacity (T-AOC) and activity of superoxide dismutase (SOD) and glutathione (GSH) was down-regulated in brain tissue, the content of homocysteine and free fatty acids (FFA) in serum of the mice. Meanwhile, AS-IV could reverse the above phenomenon, however, it does not affect the motor function of mice. AS-IV reversed these changes and it may be a potential drug for AIS therapeutics.

Overexpression of Epsin 3 enhances migration and invasion of glioma cells by inducing epithelial­mesenchymal transition.

Epsin 3 (EPN3) expression is limited to gastric parietal cells and wounded or pathological tissue rather than normal brain tissue, and although it has been identified as an oncogene in estrogen receptor­positive breast cancer and non­small cell lung cancer, its function in cancer is poorly understood. The present study aimed to investigate the association of EPN3 expression with the clinicopathological features of patients with glioma, as well as the effects of EPN3 on glioblastoma cells and the potential molecular mechanisms for its effects on glioblastoma cell behavior. EPN3 expression was assessed by immunohistochemistry in tissue samples from 167 patients with glioma, as well as by western blotting in 5 glioblastoma cell lines. The U87 and U251 glioblastoma cell lines were used to investigate the effects of EPN3 on glioblastoma cell invasion and migration through gain and loss of EPN3 expression experiments; expression levels were further investigated by reverse transcription­quantitative polymerase chain reaction (RT­qPCR) and western blot analyses. The results demonstrated that EPN3 expression levels were upregulated in high­grade glioma tissues compared with low­grade tissues, and there were varying expression levels of EPN3 in the five glioblastoma cell lines. No significant differences were observed in EPN3 expression in relation to patient age, sex or tumor size. Overexpression of EPN3 promoted glioblastoma cell migration and invasion, which we hypothesized was through affecting epithelial­mesenchymal transition (EMT). RT­qPCR and western blotting revealed that EPN3 upregulation increased the expression of Notch1 intracellular domain, ß­catenin, Slug, Twist and zinc­finger E­box­binding homeobox (ZEB)­1. These results suggested that EPN3 enhances the migration and invasion of glioblastoma cells by activating the transcription factors Slug, Twist and ZEB1, but not Snail 1 or ZEB2, to induce EMT in glioma cells; EPN3 involvement in the Notch and WNT/ß­catenin signaling pathways may contribute to this process.

Analysis of intestinal microbial communities of cerebral infarction and ischemia patients based on high throughput sequencing technology and glucose and lipid metabolism.

Currently, cerebral infarction (CI) is the leading cause of disability and the second leading cause of mortality in China, seriously affecting patient quality of life. Ischemia (IS) is considered to be the early stage of CI. The present study aims to investigate the variation of intestinal microbial communities in patients with CI and IS using high throughput sequencing technology, and then analyze the results to identify a novel potential pathogenic mechanism of CI and IS. In total, 8 patients with CI, 2 patients with IS and 10 healthy volunteers as a control were selected. Throughput sequencing technology was used to analyze the character and microbial population of the gut. The abundance of Escherichia, Bacteroides, Megamonas, Parabacteroides, Akkermansia, Prevotella, Faecalibacterium, Dialister, Bifidobacterium and Ruminococcus was the significant difference in the intestinal microbial communities of the CI and IS patients compared with the healthy group. It was also observed that CI and IS were closely associated with internal glucose metabolism. The intestinal gut disturbance of CI patients may be one of the causes inducing CI by glucose metabolism and maybe considered as a potential method to predict the disease.

MicroRNA-613 regulates the expression of brain-derived neurotrophic factor in Alzheimer's disease.

Alzheimer's disease (AD) is a neurodegenerative disorder characterized by progressive loss of memory and other cognitive functions and presents an increasing clinical challenge in terms of diagnosis and treatment. Brain-derived neurotrophic factor (BDNF) plays an important role in neuronal survival and proliferation. In the present study, the mRNA and protein expression level of BDNF was detected in serum, and cerebrospinal fluid (CSF) of patients with mild cognitive impairment (MCI), dementia of Alzheimer's type (DAT), and hippocampus in APP/PS1 mice. A significant decrease of BDNF mRNA and protein expression was observed in serum and CSF of patients and hippocampus in APP/PS1 mice compared with the corresponding controls. miR-613, which is predicted to target the 3'-UTR of BDNF, was also detected in patients and the mouse model. Opposite to the alteration of BDNF, miR-613 expression in serum, CSF and hippocampus were obviously increased compared to the controls. In conclusion, these findings showed that miR-613 may function in the development of AD and may provide new insights in diagnosis and treatment of AD.

The Effects and Molecular Mechanisms of MiR-106a in Multidrug Resistance Reversal in Human Glioma U87/DDP and U251/G Cell Lines.

Chemotherapy resistance is one of the major obstacles to effective glioma therapy. Currently, the mechanism underlying chemotherapy resistance is unclear. A recent study showed that miR-106a is an important molecule involved in chemotherapy resistance. To explore the effects and mechanisms of miR-106a on multidrug resistance reversal in human glioma cells, we silenced miR-106a expression in the cisplatin-resistant U87 (U87/DDP) and the gefitinib-resistant U251 (U251/G) glioma cell lines and measured the resulting drug sensitivity, cell apoptosis rate and rhodamine 123 content. In addition, we detected decreased expression of P-glycoprotein, MDR1, MRP1, GST-π, CDX2, ERCC1, RhoE, Bcl-2, Survivin and Topo-II, as well as reduced production of IL-6, IL-8 and TGF-ß in these cell lines. Furthermore, we found decreased expression of p-AKT and transcriptional activation of NF-κB, Twist, AP-1 and Snail in these cell lines. These results suggest that miR-106a is a promising therapeutic target for the treatment of human multidrug resistant glioma.