Construction of Multiple DNAzymes Driven by Single Base Elongation and Ligation for Single-Molecule Monitoring of FTO in Cancer Tissues.

Fat mass and obesity-associated proteins (FTO) play an essential role in the reversible regulation of N6-methyladenosine (m6A) epigenetic modification, and the overexpression of FTO is closely associated with the occurrence of diverse human diseases (e.g., obesity and cancers). Herein, we demonstrate the construction of multiple DNAzymes driven by single base elongation and ligation for the single-molecule monitoring of FTO in cancer tissues. When target FTO is present, the m6A-RNA is specifically demethylated and subsequently acts as a primer to combine with the padlock probe, initiating single-base elongation and ligation reaction to generate a closed template probe. Upon the addition of phi29 DNA polymerase, a rolling circle amplification (RCA) reaction is initiated to produce large numbers of Mg2+-dependent DNAzyme repeats. Subsequently, the DNAzymes cyclically digest the signal probes, liberating numerous Cy5 molecules that can be precisely counted by single-molecule imaging. Taking advantage of the sequence specificity of the polymerase/ligase-mediated gap-filling and ligation as well as the high amplification efficiency of RCA, this biosensor shows excellent specificity and high sensitivity with a detection limit of 5.96 × 10-16 M. It can be applied to screen FTO inhibitors and quantify FTO activity at the single-cell level. Moreover, the proposed strategy can accurately distinguish the FTO expression level in tissues of healthy individuals and breast cancer patients, providing a new platform for drug discovery, m6A modification-related research, and clinical diagnostics.

Markers of right ventricular dysfunction predict 30-day adverse prognosis of pulmonary embolism on pulmonary computed tomographic angiography.

To investigate the value of parameters of the pulmonary artery and right ventricular function in predicting the 30-day poor prognosis of patients with acute pulmonary embolism (APE). The heart rate, respiratory rate, systolic blood pressure, Wells score for APE, history of recent operation or immobilization, history of cancer, respiratory failure, smoking were significantly (P < .05) different among the control, good prognosis, and poor prognosis groups. The maximal short diameter of the right and left ventricle (RVD/LVD) ratio (P < .001) and left pulmonary artery (LPA) (P = .01) were significantly different between the good and poor prognosis groups. Systolic blood pressure (odds ratio [OR]: 0.98, P = .045) and the RVD/LVD ratio (OR: 12.57, P = .02) were significant independent risk factors for poor prognosis. The risk for poor prognosis significantly increased when the RVD/LVD ratio was >1.11 (cutoff value) with the area under the curve (AUC) of 0.71 (95% confidence interval [CI]: 0.61-0.80, P < .001). LPA (OR: 9.12, P = .01) and RVD/LVD (OR: 4.62, P = .012) were the significant independent risk factors for poor prognosis in the central pulmonary embolism. The LPA of 2.1 cm had the highest predictive value for poor prognosis in the central APE (AUC: 0.68; sensitivity 84.6%; specificity 53.1%). The RVD/LVD ratio and systolic blood pressure are significant risk factors for short-term prognosis in patients with APE. When the LPA is >2.1 cm in the central APE or the RVD/LVD is >1.11, the risk of poor prognosis increases, which can be used as important indicators for predicting the prognosis of patients with APE. Two hundred forty-three APE patients and 61 patients without APE who underwent computed tomographic pulmonary angiography (CTPA) were retrospectively enrolled as the experimental and the control group, respectively. APE patients who were followed up at the 30-day time point were divided into the good prognosis (n = 195) and poor prognosis group (n = 32). The main pulmonary artery (MPA) to the aorta (AO) ratio, maximal diameter of the LPA and right pulmonary artery (RPA), ratio of the RVD/LVD and the height and volume of the pulmonary artery (PAh and PAV, respectively) were analyzed after indexing to the body surface area.

Transcriptionally amplified synthesis of fluorogenic RNA aptamers for label-free DNA glycosylase assay.

We demonstrate for the first time the utilization of fluorogenic RNA aptamers for label-free uracil-DNA glycosylase (UDG) assay. Through rationally engineering the transcription machine with dU substitution, this assay requires only a single probe to simultaneously sense and amplify the UDG signal, achieving a low detection limit of 6.3 × 10-6 U mL-1. Moreover, it can be applied for screening UDG inhibitors and measuring endogenous UDG activity in different cells.

Normal diameters of abdominal aorta and common iliac artery in middle-aged and elderly Chinese Han people based on CTA.

To investigate the normal diameter of the abdominal aorta and common iliac arteries of the middle-aged and elderly people in China and the relationship of the diameters with age, sex, height, weight, body mass index (BMI), and body surface area (BSA). This retrospective study enrolled 625 patients including 380 males and 245 females aged 60.00 years (interquartile range 13.00 years). All clinical data and the diameters of the abdominal aorta and common iliac arteries were analyzed. The diameter of the abdominal aorta was 21.49 ± 2.49 mm at the proximal, 16.94 (interquartile range 2.39) mm at the middle, and 15.65 (interquartile range 2.90) mm at the distal segment. The diameter of the common iliac artery was 10.76 (interquartile range 1.99) mm at the right proximal, 10.41 (interquartile range 2.05) mm at the left proximal, 10.74 (interquartile range 2.25) mm at the right distal, and 10.67 (interquartile range 2.22) mm at the left distal segment. The height, weight, BSA, BMI, diameters of the proximal, middle and distal abdominal aorta as well as the proximal and distal left and right common iliac arteries were significantly higher in males than those in females (P < .001). Height, weight, BSA, and BMI were significantly (P < .001) positively correlated with the diameter of the abdominal aorta and common iliac artery at the proximal, middle, and distal segments. The middle and distal diameters of the abdominal aorta were significantly higher in males than those in females (P < .05). The diameter of the abdominal aorta at the proximal, middle, and distal segment as well as the diameter of the left and right common iliac artery at the distal segment were significantly (P < .05) increased with age. The normal values of the diameter of the abdominal aorta and common iliac arteries are suggested for the middle-aged and elderly Chinese people for clinical reference. The diameters are gender related and significantly positively correlated with BSA, height, weight, and BMI, which is beneficial for the diagnosis and treatment planning of relevant vascular diseases.

Calcium phosphate-based nanomedicine mediated CRISPR/Cas9 delivery for prostate cancer therapy.

Introduction: Erythropoietin producing hepatocyte receptor A2 (EphA2) is widely presented in the tumor cells, closely related to tumor cell migration, not cell apoptosis and proliferation. Based on its high expression in castration-resistant prostate cancer (CRPC), we herein develop a CRISPR-Cas9-based genome-editing nanomedicine to target erythropoietin producing hepatocyte receptor A2 for the treatment of castration-resistant prostate cancer. Methods: To this end, TAT was designed to stabilize the distribution of calcium, and then bound to ribonucleoprotein (RNP) to form nanoparticles RNP@CaP-TAT. Results: This nanoparticle has a simple synthesis process with good biocompatible, to achieve the knockout of tumor cells (PC-3) targeting erythropoietin producing hepatocyte receptor A2 gene and to effectively suppress the migration of tumor cells. Discussion: This delivery genome editing system provides a promising gene therapy strategy for the treatment of castration-resistant prostate cancer, showing good potential against castration-resistant prostate cancer tumor metastasis. In addition, it can be extended to other types of cancer with highly heterogeneous gene expression.

The Efficacy of Vitamin D Supplementation for Migraine: A Meta-Analysis of Randomized Controlled Studies.

INTRODUCTION: The efficacy of vitamin D for migraine remains controversial. We conduct a systematic review and meta-analysis to explore the influence of vitamin D versus placebo on treatment in migraine patients. METHODS: We search PubMed, EMbase, Web of Science, EBSCO, and Cochrane library databases through April 2020 for randomized controlled trials assessing the effect of vitamin D versus placebo on treatment efficacy in migraine patients. This meta-analysis is performed using the random-effect model. RESULTS: Five randomized controlled trials are included in the meta-analysis. Overall, compared with control group in migraine patients, vitamin D treatment is associated with substantially reduced number of headache days (standard mean difference [SMD], -0.53; 95% confidence interval [CI], -0.83 to -0.23; P = 0.0006), frequency of headache attacks (SMD, -1.09; 95% CI, -1.86 to -0.32; P = 0.006), headache severity (SMD, -0.55; 95% CI, -0.91 to -0.19; P = 0.0003), and Migraine Disability Assessment score (SMD, -0.76; 95% CI, -1.11 to -0.40; P < 0.0001). CONCLUSIONS: Vitamin D treatment is effective to alleviate migraine.

Internet-based cognitive-behavioural therapy for insomnia (ICBT-i): a meta-analysis of randomised controlled trials.

OBJECTIVE: To evaluate the effectiveness of internet-based cognitive-behavioural therapy for insomnia (ICBT-i) in adults. DESIGN: A meta-analysis of ICBT-i. DATA SOURCES: Systematic searches of randomised controlled trials of ICBT-i were performed in the PubMed, EMBASE, PsycINFO and Cochrane Library databases up to 19 June 2016. REVIEW METHOD: 2 reviewers independently performed study selection, quality assessment and data extraction. Outcomes of interest included sleep onset latency (SOL), total sleep time (TST), sleep efficiency (SE), wake after sleep onset (WASO), number of nocturnal awakenings (NWAK), and Insomnia Severity Index (ISI). RevMan 5.2 and Stata 13.0 meta-analysis software were used to perform statistical analysis. RESULTS: 14 records for 15 studies (1013 experimental group participants, 591 waiting list group participants) were included. The meta-analysis indicated that, at the post-test time point, SOL decreased by 18.41 min (95% CI 13.60 to 23.21), TST increased by 22.30 min (95% CI 16.38 to 28.23), SE increased by 9.58% (95% CI 7.30% to 11.85%), WASO decreased by 22.31 min (95% CI 13.50 to 31.11), NWAK decreased by 0.52 (95% CI 0.28 to 0.76), and ISI decreased by 5.88 points (95% CI 4.29 to 7.46). Additionally SOL, TST, SE, and WASO exhibited statistically significant improvements at follow-up versus before treatment. CONCLUSIONS: ICBT-i is an effective treatment for adults with insomnia. This conclusion should be verified in further studies.

Internet-Based Cognitive Behavioral Therapy for Insomnia (ICBT-i) Improves Comorbid Anxiety and Depression-A Meta-Analysis of Randomized Controlled Trials.

As the internet has become popularized in recent years, cognitive behavioral therapy for insomnia (CBT-i) has shifted from a face-to-face approach to delivery via the internet (internet-based CBT-i, ICBT-i). Several studies have investigated the effects of ICBT-i on comorbid anxiety and depression; however, the results remain inconclusive. Thus, a meta-analysis was conducted to determine the effects of ICBT-i on anxiety and depression. Electronic databases, including PubMed, EMBASE, PsycINFO and the Cochrane Library (throughout May 28, 2015), were systematically searched for randomized controlled trials (RCTs) of ICBT-i. Data were extracted from the qualified studies and pooled together. The standardized mean difference (SMD) and 95% confidence interval (95% CI) were calculated to assess the effects of ICBT-i on comorbid anxiety and depression. Nine records that included ten studies were ultimately qualified. The effect sizes (ESs) were -0.35 [-0.46, -0.25] for anxiety and -0.36 [-0.47, -0.26] for depression, which were stable using a between-group or within-group comparison and suggest positive effects of ICBT-i on both comorbid disorders. Although positive results were identified in this meta-analysis, additional high-quality studies with larger sample sizes are needed in the future.

Alpha lipoic acid protects lens from H(2)O(2)-induced cataract by inhibiting apoptosis of lens epithelial cells and inducing activation of anti-oxidative enzymes.

OBJECTIVE: To determine whether alpha lipoic acid (LA) can effectively protect lenses from hydrogen peroxide (H2O2)-induced cataract. METHODS: Lens from adult Sprague-Dawley rats were cultured in 24-well plates and treated without or with 0.2 mM of H2O2, 0.2 mM of H2O2 plus 0.5 mM, 1.0 mM, or 2.0 mM of LA for 24 h. Cataract was assessed using cross line grey scale measurement. Superoxide dismutase (SOD), glutathione (GSH-Px), lactate dehydrogenase (LDH), and malondialdehyde (MDA) activity or level in lens homogenates was measured. Apoptosis of lens epithelial cells in each group were detected by Terminal Deoxynucleotidyl Transferase dUTP Nick End Labeling (TUNEL) Assay. RESULTS: A total of 0.2 mM of H2O2 induced obvious cataract formation and apoptosis in lens' epithelial cells, but 0.5-2.0 mM of LA could block the effect of 0.2 mM H2O2 in inducing cataract and apoptosis. Furthermore, 0.2 mM of H2O2 significantly decreased SOD, GSH-Px, and LDH activity and significant increased MDA level in the lens, but 0.5-2.0 mM of LA blocked the effect of 0.2 mM H2O2. One mM of LA was found to be the most effective. CONCLUSIONS: LA can protect lens from H2O2-induced cataract. LA exerts protective effects through inhibition of lens' epithelial cell apoptosis and activation of anti-oxidative enzymes.