A new strategy for choosing “Q-markers” via network pharmacology,application to the quality control of a Chinese medical preparation

Due to its chemical complexity, proper quality control for a Chinese medical preparation (CMP) has been a great challenge. Choosing the appropriate quality markers (Q-markers) for quality control of CMP is an important work. Best of all, the chosen Q-markers are the main chemical compounds from the herbals as well as the active constituents of this CMP. Only in this way the established quality control system can really achieve the purpose of controlling the quality of CMP and ensuring the safely and effectively use of CMP. To achieve the purpose, network pharmacology combined with the contents of chemical compounds in the CMP has been used in this research. We took an anti-arrhythmic CMP, Shenxian-Shengmai oral liquid (SSOL), as an example. Firstly, UPLC-QTOF-MS/MS method was used to analyze the main components of SSOL. A total of 64 compounds were unambiguously or tentatively identified and 32 of them were further validated by reference compounds. Secondly, the network was constructed based on the identified compounds to predict the effective compounds related to cardiac arrhythmias. Based on the existing database and the operation method of topology, a method of double network analysis (DNAA) was proposed, from which 10 important targets in the pathway of arrhythmia were screened out, and 26 compounds had good antiarrhythmic activity. Based on the prediction results of network pharmacology along with the contents of the compounds in this CMP, ten representative compounds were chosen as the Q-markers for the quality control of SSOL. We find that five of these ten compounds, including danshensu, rosmarinic acid, salvianolic acid A, epimedin A and icariin, have antiarrhythmic activity. Then, the UPLC-DAD method was established as the control method for SSOL.     

Delivery of radix ophiopogonis polysaccharide via sucrose acetate isobutyrate-based in situ forming systems alone or combined with its mono-PEGylation

This work aimed to achieve long-lasting delivery of radix ophiopogonis polysaccharide (ROP) by sucrose acetate isobutyrate (SAIB)-based in situ forming systems (ISFSs) alone or combined with mono-PEGylation of ROP. When the ‘90%SAIB/10% solvent’ system was used, the mean residence time (MRT) of ROP was prolonged by 4.3 5?～?7.00 times and the initial release rate was reduced significantly. However, this system was only suitable for days-long sustained release of ROP in short-term therapy. As to the ‘SAIB/additives/solvent’ system containing mono-PEGylated ROP, the results indicated that SAIB/poly(d,l-lactide-co-glycolide) (PLGA)/N-methyl-2-pyrrolidone (NMP) was superior to SAIB/polylactic acid (PLA)/NMP and SAIB/PLA/ethanol in controlled release. Moreover, weeks- to months-long (16–60 d) smooth release of ROP could be achieved by varying the concentration (10–30%) and molecular weight (MW) of PLGA (10–50?kDa) or by employing a moderate MW of PEGylated ROP (～20 or ～30?kDa). With further increasing the conjugate MW to ～40?kDa, the contribution of drug elimination to its plasma retention seemed to surpass that of the SAIB-based system, resulting in that the system no longer had an obvious influence on the in vivo behavior of the conjugate. Besides, the results of host response confirmed that with less solvent being used, the SAIB-based systems showed a higher biocompatibility than the PLGA-based systems, suggesting that they could be freely chosen in the prevention and/or cure of chronic diseases.     

百草枯对A549细胞中解整合素-金属蛋白酶17表达的影响

目的构建体外百草枯（paraquat,PQ）细胞纤维化模型,观察PQ对A549细胞中解整合素-金属蛋白酶17（ADAM17）表达的影响,探讨ADAM17在PQ中毒致肺纤维化中的作用。方法体外培养A549细胞,分为正常对照组、不同浓度PQ组,应用CCK-8检测细胞活力,筛选PQ浓度和时间,显微镜下观察细胞形态,ELISA测定各组纤维化标志物I型胶原（type I collagen,Col I）和纤连蛋白（fibronectin,FN）的表达,建立细胞纤维化模型;免疫细胞化学检测A549细胞中ADAM17的分布情况,RT-PCR和Western blot分别半定量检测ADAM17 mRNA及蛋白水平的表达情况。结果（1）随着PQ浓度增加及作用时间延长,A549细胞活力呈下降趋势,差异有统计学意义（P〈0.05）,呈剂量依赖性和时间依赖性。（2）正常的A549细胞融合呈铺路石样生长,排列比较紧密,经PQ诱导后细胞排列较松散,细胞间连接变疏松,部分细胞溶解、死亡。（3）ELISA显示,随PQ浓度增加,Col I和FN表达增强,差异有统计学意义（P〈0.05）;随PQ时间延长,Col I和FN表达也逐渐增强,差异有统计学意义（P〈0.05）,成功建立PQ细胞纤维化模型。（4）免疫细胞化学显示,ADAM17在A549细胞胞浆表达。（5）RT-PCR和Western blot表明,随着PQ浓度增加,ADAM17 mRNA及蛋白水平表达明显增加,差异有统计学意义（P〈0.05）,以PQ 200 umol/L时最为明显。随着PQ作用时间延长,ADAM17 mRNA及蛋白表达水平也明显增加,差异有统计学意义（P〈0.05）,在24 h达到高峰。结论百草枯可引起肺泡上皮细胞形态学改变,导致细胞损伤,成功建立细胞的纤维化模型,对A549细胞的毒性作用具有剂量和时间依赖性。ADAM17在PQ诱导的A549细胞中过表达,可能参与了百草枯诱导的肺纤维化过程。     

SimPEL: Simulation‐based power estimation for sequencing studies of low‐prevalence conditions

Power estimations are important for optimizing genotype‐phenotype association study designs. However, existing frameworks are designed for common disorders, and thus ill‐suited for the inherent challenges of studies for low‐prevalence conditions such as rare diseases and infrequent adverse drug reactions. These challenges include small sample sizes and the need to leverage genetic annotation resources in association analyses for the purpose of ranking potential causal genes. We present SimPEL, a simulation‐based program providing power estimations for the design of low‐prevalence condition studies. SimPEL integrates the usage of gene annotation resources for association analyses. Customizable parameters, including the penetrance of the putative causal allele and the employed pathogenic scoring system, allow SimPEL to realistically model a large range of study designs. To demonstrate the effects of various parameters on power, we estimated the power of several simulated designs using SimPEL and captured power trends in agreement with observations from current literature on low‐frequency condition studies. SimPEL, as a tool, provides researchers studying low‐frequency conditions with an intuitive and highly flexible avenue for statistical power estimation. The platform‐independent “batteries included” executable and default input files are available at https://github.com/precisionomics/SimPEL .