A novel three-round multiplex PCR for SNP genotyping with next generation sequencing.

Owing to the high throughput and low cost, next generation sequencing has attracted much attention for SNP genotyping application for researchers. Here, we introduce a new method based on three-round multiplex PCR to precisely genotype SNPs with next generation sequencing. This method can as much as possible consume the equivalent amount of each pair of specific primers to largely eliminate the amplification discrepancy between different loci. After the PCR amplification, the products can be directly subjected to next generation sequencing platform. We simultaneously amplified 37 SNP loci of 757 samples and sequenced all amplicons on ion torrent PGM platform; 90.5 % of the target SNP loci were accurately genotyped (at least 15×) and 90.4 % amplicons had uniform coverage with a variation less than 50-fold. Ligase detection reaction (LDR) was performed to genotype the 19 SNP loci (as part of the 37 SNP loci) with 91 samples randomly selected from the 757 samples, and 99.5 % genotyping data were consistent with the next generation sequencing results. Our results demonstrate that three-round PCR coupled with next generation sequencing is an efficient and economical genotyping approach. Graphical Abstract The schematic diagram of three-round PCR.

Eucommia ulmoides Oliv. Leaf Extract Improves Erectile Dysfunction in Streptozotocin-Induced Diabetic Rats by Protecting Endothelial Function and Ameliorating Hypothalamic-Pituitary-Gonadal Axis Function.

Erectile dysfunction (ED) is a major complication of diabetes mellitus. Eucommia ulmoides Oliv. is used as a traditional medicine for male impotence, but no systematic study has examined its effect on diabetes-associated ED. In this study, we investigated the effects of Eucommia ulmoides Oliv. leaf extract (EULE) on restoring erectile function in streptozotocin (STZ)-induced diabetic rats model. After 16 weeks of treatment, EULE administration had significantly increased intracavernosal pressure, nitric oxide (NO) levels, and cyclic guanosine monophosphate (cGMP) concentrations. Serum superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) levels were markedly higher and serum malondialdehyde (MDA) levels were lower in the EULE-treated groups than in the diabetic model group. EULE restored NO biosynthesis by significantly increasing protein kinase B (Akt) and endothelial NO synthase (eNOS) activation. Furthermore, EULE is likely to benefit the hypothalamic-pituitary-gonadal (HPG) axis, as it increased gonadotropin-releasing hormone (GnRH), follicle-stimulating hormone (FSH), luteinizing hormone (LH), and testosterone (T) concentrations as well as hormone receptors Gnrhr, Fshr, and Lhr expression levels. Hence, EULE attenuates oxidative stress, increases NO production, and activates the Akt-eNOS pathway to restore endothelial function; moreover, EULE enhances the HPG axis to improve erectile function. These results suggest that EULE may represent a new therapeutic avenue for diabetes-associated ED.

Association of learning and living style with dyslexia in school-aged children / 中国学校卫生

Objective@#To investigate the association between the learning and living style with developmental dyslexia in school-aged children.@*Methods@#Using stratified cluster sampling, a total of 11 668 schoolaged children (grade 2 to 6) in the cities of Wuhan, Hangzhou and Jining were selected to participate in this programme from April 2017 to April 2018. The investigation tools combined the questionnaire on associated factors for reading ability, Dyslexia Checklist for Chinese Children and Pupil Rating Scale Revised Screening for Learning.@*Results@#Pupils with more than 20 minutes of exercise each day (OR=0.43-0.64) and at least 1-2 times per week (OR=0.34-0.48) had a lower risk of dyslexia. The association was observed between going to the library more than 1-2 times per semester (OR=0.41-0.62) and the decrease risk of dyslexia. Lacking active learning (OR=7.76, 95%CI=4.71-12.78), scheduled reading time (OR=2.55, 95%CI=2.01-3.23) and extracurricular training classes (OR=1.62, 95%CI=1.27-2.07) were positively associated with dyslexia. There was no significant difference in screen time duration between dyslexic and non-dyslexic children. Using electronic devices for learning was associated with decreased risk of dyslexia (OR=0.47, 95%CI=0.33-0.67), while playing video games was correlated with increased risk of dyslexia (OR=1.67, 95%CI=1.16-2.41).@*Conclusion@#Physical exercise, good study habits and using the electronic products in a proper way could reduce the risk of dyslexia to a certain extent. Parents and teachers should guide the school-aged children to develop a good learning and living style.