A Novel miRNA Detection Method Using Loop-Mediated Isothermal Amplification.

A novel ligation-based loop-mediated isothermal amplification has been developed for miRNA detection. Two stem-loop structure DNA linker A/B probes which hybridized with miRNA were designed to establish a rapid and ultrasensitive miRNA-LAMP system for miRNA detection. Target miR-200a was used to template the ligation of Linker A/B probes with SplintR Ligase and used as a dumbbell-shaped amplicon. By adding BIP/FIP and Bst 2.0 DNA polymerase, the LAMP reaction was carried out, which brought greatly improved amplification efficiency. The double-stranded DNA fluorescent dye EvaGreen was added for the detection of amplification product to achieve the quantification of the target miRNA. This method can detect miRNA in a linear range of seven orders of magnitude, with a detection limit of 100 fM. Therefore, this ultrasensitive miRNA-LAMP assay provides a new path for the highly sensitive quantitative analysis of miRNA, thereby bringing convenience to clinical diagnosis and prognostic research.

[Discussion about risk and management of Chinese patent medicine with double identity].

Chinese patent medicine with double identity was a special phenomenon, and many preparations not only were prescription drugs but also over the counter ( OTC) drugs, which brought a lot of trouble. Based on statistics of list of OTC medicines of CFDA, related varieties, route of administration and functions of these drugs were searched. The causes of insufficient were analyzed and the potential risk was investigated. To ensure the safety of drug usage for the patient, risk management system should be set up by improving the technical requirements for registration, improving the drug labels and manuals, playing the role of pharmacists in pharmacy services and raising awareness of doctor and patient for these drugs.

[Simultaneous determination of phenolic acids in Guanxinning injection by HPLC].

This paper was aim to determine five phenolic acids, sodium danshensu (SD), protocatechuic aldehyde (PA), rosmarinic acid (RA), lithospermic acid (LA) and salvianolic acid B (SAB), in Guanxinning injection. In the test, Kromasil C18 column (4.6 mm x 250 mm, 5 µm) was adopted, with acetonitrile-3% formic acid solution as the mobile phase for gradient elution. The flow rate was 1 mL · min, the column temperature was 30 °C and the detection wavelength was 280 nm. According to the results of the test, SD, PA, RA, LA and SAB showed good linear relations between peak areas and sample sizes in 0.006 06-4.04 (r = 0.999 3), 0.006 15-4.10 (r = 0.999 4), 0.005 94-3.96 (r = 0.999 3), 0.006 06-4.04(r = 0.999 1) and 0.006 09-4.06 (r = 0.999 2) µg, respectively. The average recoveries (n = 6) were 98.9% (RSD 0.75%), 98.1% (RSD 1.2%), 100% (RSD 0.77%), 98.7% (RSD 1.7%), 102% (RSD 0.68%), respectively. The above 5 components were determined in 13 batches of samples by using the established method. The method was simple, accurate and highly reproducible that it could be used for quality control of the components in Guanxinning injection.