Protective effect of the extract of Yi-Qi-Fu-Mai preparation on hypoxia-induced heart injury in mice

Yi-Qi-Fu-Mai(YQFM) is extensively used clinically to treat cardiovascular diseases in China. To explore the anti-hypoxia effect of the extract of YQFM preparation(EYQFM), the EYQFM(1.4, 2.8, and 5.5 g·kg-1·d-1) was assessed for its heart-protective effect in a chronic intermittent hypoxia(CIH) animal model(oxygen pressure 7%-8%, 20 min per day) for 28 days of treatment. Betaloc(0.151 6 g·kg^(-1)·d^(-1)) was used as a positive control. The histopathological analyses of heart in CIH mice were conducted. Several cardiac state parameters, such as left ventricular ejection fractions(EF), stroke volume(SV), expression of creatine kinase(CK), lactate dehydrogenase(LDH), superoxide dismutase(SOD), and malondialdehyde(MDA) were measured. The results showed that treatment with EYQFM markedly reversed swelling of the endothelial cells and vacuolization in the heart when compared with the model group. Further study demonstrated that EYQFM significantly improved ventricular myocardial contractility by increasing EF and SV. In addition, EYQFM inhibited the activity of CK, LDH, decreased the level of MDA and improved SOD activity. The results demonstrated that EYQFM significantly improved the tolerability of myocardium to hypoxia and ameliorated the cardiac damage in the CIH model.

Simplified methodology for a modular and genetically expanded protein synthesis in cell-free systems

Genetic code expansion,which enables the site-specific incorporation of unnatural amino acids into proteins,has emerged as a new and powerful tool for protein engineering.Currently,it is mainly utilized inside living cells for a myriad of applications.However,the utilization of this technology in a cell-free,reconstituted platform has several advantages over living systems.The typical limitations to the employment of these systems are the laborious and complex nature of its preparation and utilization.Herein,we describe a simplified method for the preparation of this system from Escherichia coli cells,which is specifically adapted for the expression of the components needed for cell-free genetic code expansion.Besides,we propose and demonstrate a modular approach to its utilization.By this approach,it is possible to prepare and store different extracts,harboring various translational components,and mix and match them as needed for more than four years retaining its high efficiency.We demonstrate this with the simultaneous incorporation of two different unnatural amino acids into a reporter protein.Finally,we demonstrate the advantage of cell-free systems over living cells for the incorporation ofδ-thio-boc-lysine into ubiquitin by using the methanosarcina mazei wild-type pyrrolysyl tRNACUA and tRNA-synthetase pair,which could not be achieved in a living cell.