OBJECTIVE Ischemic stroke remains a significant cause of death and disability in industrialized nations. Janus tyrosine kinase (JAK) and signal transducer and activator of transcription (STAT) of the JAK2/STAT3 pathway play important roles in the downstream signal pathway regulation of ischemic stroke-related inflammatory neuronal damage. Recently, microRNAs (miRNAs) have emerged as major regulators in cerebral ischemic injury; therefore, the authors aimed to investigate the underlying molecular mechanism between miRNAs and ischemic stroke, which may provide potential therapeutic targets for ischemic stroke. METHODS The JAK2- and JAK3-related miRNA (miR-135, miR-216a, and miR-433) expression levels were detected by real-time quantitative reverse-transcriptase polymerase chain reaction (qRT-PCR) and Western blot analysis in both oxygen-glucose deprivation (OGD)-treated primary cultured neuronal cells and mouse brain with middle cerebral artery occlusion (MCAO)-induced ischemic stroke. The miR-135, miR-216a, and miR-433 were determined by bioinformatics analysis that may target JAK2, and miR-216a was further confirmed by 3' untranslated region (3'UTR) dual-luciferase assay. The study further detected cell apoptosis, the level of lactate dehydrogenase, and inflammatory mediators (inducible nitric oxide synthase [iNOS], matrix metalloproteinase-9 [MMP-9], tumor necrosis factor-α [TNF-α], and interleukin-1β [IL-1β]) after cells were transfected with miR-NC (miRNA negative control) or miR-216a mimics and subjected to oxygen-glucose deprivation/reoxygenation (OGD/R) damage with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, annexin V-FITC/PI, Western blots, and enzyme-linked immunosorbent assay detection. Furthermore, neurological deficit detection and neurological behavior grading were performed to determine the infarction area and neurological deficits. RESULTS JAK2 showed its highest level while miR-216a showed its lowest level at day 1 after ischemic reperfusion. However, miR-135 and miR-433 had no obvious change during the process. The luciferase assay data further confirmed that miR-216a can directly target the 3'UTR of JAK2, and overexpression of miR-216a repressed JAK2 protein levels in OGD/R-treated neuronal cells as well as in the MCAO model ischemic region. In addition, overexpression of miR-216a mitigated cell apoptosis both in vitro and in vivo, which was consistent with the effect of knockdown of JAK2. Furthermore, the study found that miR-216a obviously inhibited the inflammatory mediators after OGD/R, including inflammatory enzymes (iNOS and MMP-9) and cytokines (TNF-α and IL-1β). Upregulating miR-216a levels reduced ischemic infarction and improved neurological deficit. CONCLUSIONS These findings suggest that upregulation of miR-216a, which targets JAK2, could induce neuroprotection against ischemic injury in vitro and in vivo, which provides a potential therapeutic target for ischemic stroke.

PurposeThe Questionnaire – Children with Difficulties (QCD) has been developed and used to evaluate daily-life problems in children during specified periods of the day. The objective of this study was to evaluate the reliability and validity of the QCD for Chinese children or adolescents with attention-deficit/hyperactivity disorder (ADHD).Patients and methodsOutpatients with ADHD aged 6–18 years who visited psychiatry clinics were enrolled at four study centers in China. Patients with severe psychiatric disorders were excluded. Parents of all enrolled patients were given the QCD, the Swanson, Nolan and Pelham IV (SNAP-IV), and the Weiss Functional Impairment Scale-Parent (WFIRS-P) questionnaires and were asked to complete all three questionnaires. The reliability of the QCD was examined by Cronbach’s alpha, which assessed the internal consistency of the questionnaire. Concurrent criterion validity of QCD scores was examined by Spearman’s correlation of QCD with SNAP-IV and WFIRS-P scores.ResultsA total of 200 Chinese patients were analyzed (average age, 10.4±2.66 years). The majority of patients were male (77.5%), and 49.0% had the combined ADHD subtype. Cronbach’s alpha for QCD was 0.88. Correlation coefficients of the QCD total score with SNAP-IV total score and WFIRS-P average score were −0.47 and −0.57, respectively. Correlations for the QCD with SNAP-IV and WFIRS-P were statistically significant (P<0.01). The area under the curve for sensitivity and specificity of the QCD compared with the SNAP-IV and WFIRS-P was 0.70 and 0.71, respectively. The ADHD severity discrimination threshold range of the QCD total score was 30–35.ConclusionOur study results found the QCD to be a reliable and valid instrument and recommend its use in clinical practice to identify and evaluate daily-life problems of ADHD patients during specified periods of the day in China.