Long Non-Coding RNA MALAT1 Promotes Acute Cerebral Infarction Through miRNAs-Mediated hs-CRP Regulation Abstract The occurrence of cerebral infarction commonly takes atherosclerosis as the pathophysiological basis, accompanied by chronic inflammation. Hypersensitive C-reactive protein (hs-CRP) is an important inflammatory factor involved in the formation of atherosclerosis. This study aims to investigate the regulation of hs-CRP expression by long-chain non-coding RNA (LncRNA) MALAT1 in acute cerebral infarction patients. Plasma levels of LncRNA MALAT1 and hsa-miR-145-5p, hsa-miR-140-5p, hsa-miR-483-3p, and hsa-miR-338-3p in 256 Chinese Han ACI patients and 256 controls were analyzed. HUVECs were transfected with LncRNA MALAT1, MALAT1 NC, and si-MALAT1, respectively. The expression levels of hsa-miR-145-5p, hsa-miR-140-5p, hsa-miR-483-3p, and hsa-miR-338-3p were analyzed. Then, HUVECs were transfected with hsa-miR-145-5p inhibitor, hsa-miR-140-5p inhibitor, hsa-miR-483-3p inhibitor, hsa-miR-338-3p inhibitor, and hsa-miR-145-5p mimic, hsa-miR-140-5p mimic, hsa-miR-483-3p mimic, hsa-miR-338-3p mimic, and the expression level of hs-CRP was detected by Western blotting. The levels of hsa-miR-145-5p, hsa-miR-140-5p, hsa-miR-483-3p, and hsa-miR-338-3p in the plasma of ACI patients were significantly lower than those in the control group (p < 0.001), and the plasma LncRNA MALAT1 levels were significantly higher in ACI patients than in the control group (p < 0.001). The level of LncRNA MALAT1 in plasma of ACI patients and control group was negatively correlated with hsa-miR-145-5p, hsa-miR-140-5p, hsa-miR-483-3p, and hsa-miR-338-3p (r = − 0.36, − 0.79, − 0.76, − 0.75; − 0.60, − 0.68, − 0.48, − 0.56). Plasma levels of hsa-miR-145-5p, hsa-miR-140-5p, hsa-miR-483-3p, and hsa-miR-338-3p were negatively correlated with hs-CRP levels in patients with ACI and controls (r = − 0.74, − 0.81, − 0.84, − 0.56; − 0.61, − 0.69, − 0.69, − 0.50). MALAT1 transfection resulted in the decreased levels of hsa-miR-145-5p, hsa-miR-140-5p, hsa-miR-483-3p, and hsa-miR-338-3p in HUVECs while overexpression of hsa-miR-145-5p, hsa-miR-140-5p, hsa-miR-483-3p, and hsa-miR-338-3p led to a decrease in hs-CRP levels in HUVECs. LncRNA MALAT1 induced the upregulation of CRP expression through inhibiting the expression of hsa-miR-145-5p, hsa-miR-140-5p, hsa-miR-483-3p, and hsa-miR-338-3p.

Age Matters: an Atypical Association Between Polymorphism of MTHFR and Clinical Phenotypes in Children with Schizophrenia Abstract Methylenetetrahydrofolate reductase (MTHFR) polymorphism may increase the risk of schizophrenia in adults and aggravate related symptoms, while it is unknown whether similar risk applies in children with schizophrenia. While average onset age of schizophrenia is between the ages of 15 and 25, there are no studies on the relationship between MTHFR polymorphism and childhood-onset schizophrenia (COS). Here, we aimed to explore the risk of MTHFR polymorphism in children and examine the effects of MTHFR polymorphism on disease onset and clinical features in the COS patients. Pediatric patients with schizophrenia (n = 97) as well as age- and sex-matched controls (n = 92) were enrolled from the pediatric department. We evaluated clinical features including disease onset age, duration, Positive and Negative Syndrome Scale (PANSS), Personal and Social Performance Scale (PSP), and Clinical Global Impression (CGI). The three major MTHFR genotypes (G1793A, C677T, and A1298C) were examined in all subjects and the association between MTHFR polymorphism and clinical features of schizophrenia was analyzed. The G1793A polymorphism and the total number of MTHFR risk alleles were associated with an increased risk of schizophrenia in children. The A1298C polymorphism contributed to prolong the duration time of schizophrenia. Inconsistent with expectations, no significant associations were found between MTHFR C677T polymorphism and schizophrenia in children. Both G1793A and multi-site MTHFR polymorphisms are associated with an increased risk of schizophrenia in children, while A1298C polymorphism contributes to prolonged disease duration. While C677T is known to play major roles in the risk of adult schizophrenia, our finding for the first time suggests an age-specific association between MTHFR polymorphisms and schizophrenia.

Inhibition of Ubc9-Induced CRMP2 SUMOylation Disrupts Glioblastoma Cell Proliferation Abstract Glioblastoma (GBM) is the most aggressive astrocytoma. Despite maximum treatment, the GBM usually recurs and the patient survival is poor. Thus, understanding the molecular mechanism of GBM progression will be meaningful to ameliorate this situation. In this study, collapsin response mediator protein 2 (CRMP2) and Ubc9 protein levels were evaluated in three GBM cell lines. Sumoylated CRMP2 were enriched and immunoprecipitated using SUMO1 and IgG antibodies. CRMP2-K374A mutant was generated by site-direct mutagenesis. All indicated constructs were transfected into GL15 cells, and the corresponding proliferation-promoting effect was assessed through cell proliferation ratio. The t-CSM peptide was used to disturb Ubc9-CRMP2 interaction. CRMP2 is expressed in all tested GBM cell lines. The Ubc9 protein levels are positively correlated with CRMP2 level, and both can promote GBM cell proliferation. Blocking CRMP2 SUMOylation through SUMOylation-incompetent mutant or small peptide suppresses CRMP2-induced GBM cell proliferation. This study demonstrates that the CRMP2 SUMOylation exists widely in GBM cells and drives glioblastoma proliferation. CRMP2 SUMOylation inhibition can significantly suppress GBM proliferation in vitro.

Effects of NLRP6 in Cerebral Ischemia/Reperfusion (I/R) Injury in Rats Abstract The NOD-like receptor protein 6 (NLRP6), an intracytoplasmic pattern recognition receptor in the nucleotide-binding domain, leucine-rich repeat-containing (NLR) innate immune receptor family, influences the inflammation reaction. The role of NLRP6 in cerebral ischemia-reperfusion (I/R) injury in rats is unclear. We explore the function of NLRP6 in cerebral I/R injury. The investigators used a middle cerebral artery occlusion/reperfusion model (MCAO) to imitate ischemic injury. We found the peak expression of NLRP6 is in 48-h post-cerebral I/R injury. The expression of NLRP6 siRNA, as well as the expression of protein and mRNA, was detected by Western blot and qRT-PCR. The degree of IL-1β and IL-18 was assessed by ELISA. After downregulating NLRP6, the expression of IL-1β, IL-18, cleaved Caspase-1, and myeloperoxidase (MPO) were reduced. In HE and Nissl staining, pathological injury of brain tissue after downregulating NLRP6 was improved. NLRP6 siRNA decreased the NLRP6-ASC binding states by CO-IP. NRP6 has a pro-inflammatory effect in cerebral I/R injury, which may provide a new target for the treatment of cerebral I/R injury.

The Impact of Morphine on Reproductive Activity in Male Rats Is Regulated by Rf-Amid-Related Peptide-3 and Substance P Adjusting Hypothalamic Kisspeptin Expression Abstract Obviously, opiates (e.g., morphine) are associated with the suppression and dysfunction of reproductive axis. It has been reported that substance P (SP) and RF-amid-related peptide-3 (RFRP-3) can exhibit anti-opioid effects in some regions of the nervous system. Moreover, SP and RFRP-3 are deemed as neuropeptides which exert modulatory and regulatory impacts on the function of the reproductive axis. The precise interactions of morphine with SP or RFRP-3 on the parameters of the reproductive activity, however, are not fully known. The present study was aimed to determine the impacts of the interaction of morphine either with SP or RFRP-3 on the hormonal and behavioral parameters of reproductive activity in male rats. In addition, it was aimed at determining whether the effects of these interactions rely on kisspeptin/G protein coupled receptor 54 (GPR54) pathway as the main upstream pulse generator and the mediator of the function of many inputs of gonadotropin-releasing hormone (GnRH)/luteinizing hormone (LH) system or not. Altogether, the resulted data from the sexual behavior tests, radioimmunoassay of LH/testosterone, and real-time quantitative PCR for the assessment of the expression of hypothalamic Kiss1, Gpr54, and Gnrh1 genes following concomitant administration of morphine with SP or RFRP-3 revealed that the suppressing effects of morphine on the parameters of reproductive axis activity can be affected by the administration of either RFRP-3 or SP. It is advocated that SP and RFRP-3, by the modulation of the expression of hypothalamic Kiss1, can possibly antagonize the effects of morphine on GnRH/LH system and sexual behavior.

Cerebral MAO Activity Is Not Altered by a Novel Herbal Antidepressant Treatment Abstract Inhibition of monoamine oxidase (MAO)-A/B can ameliorate depressive- and anxiety-related symptoms via increase of monoamine extracellular levels. However, such inhibition can also instigate hypertensive response following exposure to dietary tyramine (i.e., “the cheese effect”). Novel herbal treatment (NHT) is an herbal formula that has been demonstrated to reduce depressive- and anxiety-like symptoms in pre-clinical studies. The aim of the current study was to examine whether the therapeutic potential of NHT is underlain by inhibition of MAO-A/B and whether NHT poses a risk for tyramine hyper-potentiation. Unpredictable chronic mild stress (UCMS)–exposed mice and naïve mice were treated for 3 weeks with NHT (30 mg/kg; i.p.), the selective serotonin reuptake inhibitor (SSRI) escitalopram (15 mg/kg; i.p.), or saline. Subsequently, MAO-A/B activities in the hypothalamus, striatum, and prefrontal cortex (PFC) were assessed. Exposure to UCMS led to significant increases in both MAO-A and MAO-B activities in the hypothalamus (p < 0.001) and in the PFC (p < 0.01 for MAO-A; p < 0.001 for MAO-B). Neither NHT nor escitalopram had any notable effects. Treatment with NHT was supported as safe in terms of risk for inducing a hypertensive response. The antidepressant- and anxiolytic-like effects of NHT are mediated via pathways other than MAO-A/B inhibition.

Association Between Cystatin C and the Risk of Ischemic Stroke: a Systematic Review and Meta-analysis Abstract Ischemic stroke is a disease that affects people’s health and quality of life. Cystatin C has been found as a new biomarker of cardiovascular disease. We performed this meta-analysis to assess the relationship between cystatin C and the risk of ischemic stroke. The studies on looking at the association between cystatin C and ischemic stroke were identified from inception to November 18, 2018. We performed a random-effects meta-analysis using mean difference. Nine studies with a total of 3773 ischemic stroke patients were included into the meta-analysis. Our results showed that patients with ischemic stroke had significantly higher serum cystatin C concentrations compared with the participants without ischemic stroke (pooled mean difference, 0.11; 95% confidence interval (CI), 0.00–0.22; P = 0.04), in particular acute ischemic stroke and subclinical cerebral infarction (mean difference, 0.23; 95% CI, 0.11–0.36; P = 0.0003 and mean difference, 0.07; 95% CI, 0.05–0.09; P < 0.00001, respectively). Cystatin C was associated with ischemic stroke, and it could be considered a predictor for the risk of ischemic stroke, especially in acute ischemic stroke and subclinical cerebral infarction.

Long Non-coding RNA BACE1-AS May Serve as an Alzheimer’s Disease Blood-Based Biomarker Abstract Circulating long noncoding RNAs (lncRNAs) might serve as biomarkers for different pathological conditions. BACE1-AS lncRNA upregulates in the brain of people with Alzheimer’s disease (AD) and might be detected in the bloodstream. To reveal if lncRNA BACE1-AS may serve as a blood-based biomarker for AD, we compared its levels in plasma and plasma-derived exosomes between AD (n = 45) and healthy people (n = 36). Exosomes were purified from plasma by Invitrogen™ Total Exosome Isolation Kit and characterized by scanning electron microscopy (SEM) and dynamic light scattering (DLS). Total RNA was extracted from whole plasma, and plasma-derived exosomes using TRIzol® LS or TRIzol® Reagents respectively were then reverse transcribed to the cDNA using PrimeScript II cDNA synthesis kit. The BACE1-AS levels were quantified by real-time PCR, and their biomarker potencies were evaluated using ROC curve analysis. Results obtained verified the presence of BACE1-AS in the plasma samples of both AD and healthy controls. We did not observe any significant differences between the levels of BACE1-AS in the plasma or plasma-derived exosomes of AD and control people. However, there were significant differences between AD subgroups and control in the whole plasma samples. The BACE1-AS level was low in pre-AD subgroup but it was high in full-AD people compared to the healthy controls. Moreover, ROC curve analysis revealed that lncRNA BACE1-AS may discriminate pre-AD and healthy control (75% sensitivity and 100% specificity), full-AD and healthy control (68% sensitivity and 100% specificity), and pre-AD and full-AD subgroups (78% sensitivity and 100% specificity), highlighting its potential as a biomarker for AD development. In conclusion, plasma BACE1-AS level may serve as a potent blood-based biomarker for Alzheimer’s disease.

Identification of Potential Biomarkers with Diagnostic Value in Pituitary Adenomas Using Prediction Analysis for Microarrays Method Abstract Pituitary adenomas are the most common intrasellar tumors. Patients should be identified at an early stage so that effective treatment can be implemented. The study aims at detecting the potential biomarkers with diagnostic value of pituitary adenomas. Using a total of seven gene expression profiles (GEPs) of the datasets from the Gene Expression Omnibus (GEO) database, we first screened 1980 significant differentially expressed genes (DEGs). Then, we employed the prediction analysis for microarray (PAM) algorithm to identify 340 significant DEGs able to differ pituitary tumor from normal samples, which include 208 upregulated DEGs and 132 downregulated DEGs. DAVID database was used to carry out the enrichment analysis on Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO) pathways. We found that upregulated candidates were enriched in protein folding and metabolic pathways. Downregulated DEGs saw a significant enrichment in insulin receptor signaling pathway and hedgehog signaling pathway. Based on the protein-protein interaction (PPI) network as well as module analysis, we determined ten hub genes including PHLPP, ENO2, ACTR1A, EHHADH, EHMT2, FOXO1, DLD, CCT2, CSNK1D, and CETN2 that could be potential biomarkers with diagnostic value in pituitary adenomas. In conclusion, the study contributes to reliable and potential molecular biomarkers with diagnostic value. Moreover, these potential biomarkers may be used for prognosis and new therapeutic targets for the pituitary adenomas.

Age and Sex-Related Changes to Gene Expression in the Mouse Spinal Cord Abstract The spinal cord is essential for neuronal communication between the brain and rest of the body. To gain further insight into the molecular changes underpinning maturation of the mouse spinal cord, we analysed gene expression differences between 4 weeks of age (prior to puberty onset) and adulthood (8 weeks). We found 800 genes were significantly differentially expressed between juvenile and adult spinal cords. Gene ontology analysis revealed an overrepresentation of genes with roles in myelination and signal transduction among others. The expression of a further 19 genes was sexually dimorphic; these included both autosomal and sex-linked genes. Given the presence of steroid hormone receptors in the spinal cord, we also looked at the impact of two major steroid hormones, oestradiol and dihydrotestosterone (DHT) on spinal cord gene expression for selected genes. In gonadectomised male animals, implants with oestradiol and DHT produced significant changes to spinal cord gene expression. This study provides an overview of the global gene expression changes that occur as the spinal cord matures, over a key period of maturation. This confirms that both age and sex are important considerations in studies involving the spinal cord.