Electroacupuncture alleviates cerebral ischemia and reperfusion injury via modulation of the ERK1/2 signaling pathway

Electroacupuncture（EA） has anti-oxidative and anti-inflammatory actions,but whether the neuroprotective effect of EA against cerebral ischemia-reperfusion（I/R） injury involves modulation of the extracellular regulated kinase 1/2（ERK1/2） signaling pathway is unclear.Middle cerebral artery occlusion（MCAO） was performed in Sprague-Dawley rats for 2 hours followed by reperfusion for 24 hours.A 30-minute period of EA stimulation was applied to both Baihui（DU20） and Dazhui（DU14） acupoints in each rat（10 mm EA penetration depth,continuous wave with a frequency of 3 Hz,and a current intensity of 1–3 m A） when reperfusion was initiated.EA significantly reduced infarct volume,alleviated neuronal injury,and improved neurological function in rats with MCAO.Furthermore,high m RNA expression of Bax and low m RNA expression of Bcl-2 induced by MCAO was prevented by EA.EA substantially restored total glutathione reductase（GR）,glutathione（GSH） and glutathione peroxidase（GSH-Px） levels.Additionally,Nrf2 and glutamylcysteine synthetase（GCS） expression levels were markedly increased by EA.Interestingly,the neuroprotective effects of EA were attenuated when ERK1/2 activity was blocked by PD98059（a specific MEK inhibitor）.Collectively,our findings indicate that activation of the ERK1/2 signaling pathway contributes to the neuroprotective effects of EA.Our study provides a better understanding of the regulatory mechanisms underlying the therapeutic effectiveness of EA.

Comparative studies of two methods for miRNA isolation from milk whey

Micro RNAs（mi RNAs） from milk whey have been considered for their potential as noninvasive biomarkers for milk quality control and disease diagnosis. However, standard protocols for mi RNA isolation and quantification from milk whey are not well established. The objective of this study was to compare two methods for the isolation of mi RNAs from milk whey. These two methods were modified phenol-based technique（Trizol LS followed by phenol precipitation, the TP method） and combined phenol and column-based approach（Trizol LS followed by cleanup using the mi RNeasy kit, the TM method）. Yield and quality of RNA were rigorously measured using a Nano Drop ND-1000 spectrophotometer and then the distribution of RNA was precisely detected in a Bioanalyzer 2100 instrument by microchip gel electrophoresis. Several endogenous mi RNAs（bta-mi R-141, bta-mi R-146 a, bta-mi R-148 a, bta-mi R-200 c, bta-mi R-362, and bta-mi R-375） and an exogenous spike-in synthetic control mi RNA（cel-mi R-39） were quantified by real-time polymerase chain reaction（PCR） to examine the apparent recovery efficiency of milk whey mi RNAs. Both methods could successfully isolate sufficient small RNA（200 nt） from milk whey, and their yields were quite similar. However, the quantification results show that the total mi RNA recovery efficiency by the TM method is superior to that by the TP method. The TM method performed better than the TP for recovery of milk whey mi RNA due to its consistency and good repeatability in endogenous and spike-in mi RNA recovery. Additionally, quantitative recovery analysis of a spike-in mi RNA may be more accurate to reflect the milk whey mi RNA recovery efficiency than using traditional RNA quality analysis instruments（Nano Drop or Bioanalyzer 2100）.