The development of peptides and proteins is hampered by their rapid clearance in liver and other body tissues by proteolytic enzymes, so these drugs are difficult to administer except for the injection. Here, we designed and fabricated a novel biodegradable and redox-responsive submicron capsules through the layer-by-layer technique with poly(L-aspartic acid) and chitosan for transmucosal delivery of proteins and peptides. TEM graphs reveal that the intact submicron capsules were obtained and the shell of submicron capsules was about 40 nm. The mucoadhesion test indicates that the adsorption amount of the mucin could achieve up to 96.2 μg per 2 mg. The cell viability test shows that all types of submicron capsules had good cytocompatibility and the cell viability was above 90 %. As a drug model, the insulin could be loaded in the submicron capsules, and the loading efficiency was about 5 %. The release amount of insulin could be regulated by the levels of GSH. Therefore, the mucoadhesive submicron capsules as vehicles have a potential for the mucosal delivery (e.g. nasal and buccal) of therapeutic peptide and protein drugs.
Loop-mediated isothermal amplification (LAMP) is a novel nucleic acid amplification method with high specificity and rapidity under isothermal conditions. According to the LAMP method, a rapid and simple detection system was established for bovine embryo sexing. Two sets of primers were designed by targeting the bovine male-specific sequence and bovine common sequence respectively. The reaction condition of the detection system was optimized within 60 min under isothermal conditions of 65 degrees C by detection of the reaction mixture on agarose gel. Especially, the primers F2 and B2 could replace the F3 and B3 as outer primers, making the primer design simpler and the amplification efficiency higher. Additionally, codeposition of dNTPs was firstly performed to detect the reaction products by addition of 1 microl 0.1 mM CuSO(4), the visible ring-shaped deposit was found in the middle of the reaction tube with negative mixture. It could be employed as an alternative method in the detection of the reaction products in place of the time-consuming electrophoresis or the turbidity meter. Furthermore, the embryo sexing system was carried out in the embryo transfer and achieved 98% of efficiency and 99.5% of accuracy.
A crude ethanol extract of roots of SaMa przewalskii Masim was studied by micellar electrokinetic chromatography. Optimum conditions for the separation were established. Three peaks belonging to cryptotanshinone. dihydrotanshinone I and tanshinone IIA were identified by known standards. The three substances are valuable pharmaceutical ingredients of the plant.
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