Network Pharmacology-Based Dissection of the Bioactive Compounds and Pharmacological Mechanisms of Yiqi Fumai Lyophilized Injection for the Treatment of Heart Failure

Objective:Yiqi Fumai Lyophilized Injection(YQFM),a Chinese medicine injection,has been widely used for the treatment of cardiovascular diseases,especially heart failure(HF).However,bioactive compounds and underlying mechanisms of YQFM in treating HF remain poorly understood.Materials and Methods:Network pharmacology was employed to investigate the bioactive compounds and mechanisms of YQFM.A compound-target network was constructed to screen bioactive compounds based on contribution index calculation.Then,an adriamycin-induced HF rat model was established to evaluate the cardio-protective effects of YQFM by hematoxylin and eosin staining and enzyme-linked immunosorbent assays.Results:Network pharmacology indicated that YQFM may alleviate HF through 36 compounds and 109 targets.Particularly,ginsenosides Rb1,Rg1,Re,Rf,Rb2,Rh1,schisandrin,and ginsenoside Rc were indicated as the top contributors of YQFM in treating HF.YQFM was predicted to act on multiple targets such as vascular endothelial growth factor A,interleukin-2(IL-2),IL-6,and IL-1β,as well as to regulate signaling pathways such as hypoxia-inducible factor 1,tumor necrosis factor,VEGF,and PI3K-Akt.The pharmacological study suggested that YQFM could attenuate cardiac injury and up-regulate plasma concentrations of VEGFR-1 and NO in HF rats.Ginsenoside Rb1,as the major contributor from network pharmacology analysis,also showed a cardioprotective effect and up-regulation of VEGFR-1 in plasma.Conclusions:Ginsenosides and schisandrin were predicted as the most important contributors to the cardioprotective effect of YQMF.Ginsenoside Rb1 was proved to alleviate HF and increase the plasma concentration of VEGFR-1.

甲状腺未分化癌临床特征及预后分析:单中心20年回顾性研究

目的探究甲状腺未分化癌(anaplastic thyroid carcinoma,ATC)患者临床特征和预后,探索综合治疗对ATC患者预后的价值。方法回顾性分析2001—2020年中国医学科学院肿瘤医院确诊为ATC患者的临床病理资料,并根据治疗方式分为单纯手术组和综合治疗组。生存分析采用Kaplan-Meier法,多因素分析采用Cox风险回归模型。结果共纳入47例患者,男性24例,女性23例,中位年龄63岁。中位随访3.37个月,其中42例患者死亡,均因肿瘤复发进展死亡,全组中位生存时间4.33个月。单因素分析显示,治疗前出现喉返神经受累症状、远处转移、白细胞升高以及单纯手术治疗等因素与预后不良有关(P值均<0.05)。多因素分析显示,治疗前出现喉返神经受累症状(HR=2.49,95%CI:1.16~5.32,P=0.019)、远处转移(HR=2.33,95%CI:1.06~5.16,P=0.036)和白细胞升高(HR=2.50,95%CI:1.16~5.40,P=0.020)是预后不良的独立危险因素,综合治疗较单纯手术可显著改善总生存期(HR=0.22,95%CI:0.10~0.47,P<0.001)。结论ATC患者中,初诊无喉返神经受累症状、白细胞正常、无远处转移的患者预后相对好,综合治疗有助于改善患者预后。

正向×反相离线二维液相色谱-轨道质谱联用检测通脉颗粒中的微量成分

通脉颗粒是由丹参、川芎、葛根三味中药的水提物制成的用于治疗心血管疾病的复方制剂,主要成分为异黄酮苷、异黄酮及酚酸类。低极性化合物如丹参酮和酚酞类也可能对活性有贡献,但很难检测到。因此,本研究建立了正向×反相离线二维液相色谱方法对通脉颗粒中的有效成分进行富集和分离。采用正向SEP-PAK硅胶固相萃取柱分离通脉颗粒的乙酸乙酯萃取物,分别以石油醚、乙酸乙酯–甲醇(6:1,v/v)、甲醇洗脱获得三个流分,然后采用超高效液相反向色谱串联四级杆-静电场轨道阱质谱对其进行检测。葛根中黄酮类成分,丹参中酚酸类成分及川芎中酚酞类成分分布于不同流分中。从通脉颗粒中初步鉴定了131个化合物,其中葛根59个,丹参26个,川芎43个,丹参和川芎的共有成分3个。26个化合物经过对照品比对鉴定。本研究为中药复方成分的全面分析提供了有效方法。

Identification of Nonstructural Protein 8 as the N-Terminus of the RNA-Dependent RNA Polymerase of Porcine Reproductive and Respiratory Syndrome Virus

Porcine reproductive and respiratory syndrome virus (PRRSV) is a member within the family Arteriviridae of the order Nidovirales. Replication of this positive-stranded RNA virus within the host cell involves expression of viral replicase proteins encoded by two ORFs, namely ORF1 a and ORF1 b. In particular, translation of ORF1 b depends on a-1-ribosomal frameshift strategy. Thus, nonstructural protein 9 (nsp9), the first protein within ORF1 b that specifies the function of the viral RNA-dependent RNA polymerase, is expressed as the C-terminal extension of nsp8, a small nsp that is encoded by ORF1 a. However, it has remained unclear whether the mature form of nsp9 in virus-infected cells still retains nsp8,addressing which is clearly critical to understand the biological function of nsp9. By taking advantage of specific antibodies to both nsp8 and nsp9, we report the following findings. (1) In infected cells, PRRSV nsp9 was identified as a major product with a size between 72 and 95 k Da (72–95 KDa form), which exhibited the similar mobility on the gel to the in vitro expressed nsp8–9 ORF1 b, but not the ORF1 b-coded portion (nsp9 ORF1 b). (2) The antibodies to nsp8, but not to nsp7 or nsp10, could detect a major product that had the similar mobility to the 72–95 KDa form of nsp9. Moreover, nsp9 could be co-immunoprecipitated by antibodies to nsp8, and vice versa. (3) Neither nsp4 nor nsp2 PLP2 was able to cleave nsp8–nsp9 in vitro. Together, our studies provide experimental evidence to suggest that nsp8 is an N-terminal extension of nsp9.Our findings here paves way for further charactering the biological function of PRRSV nsp9.

Schaftoside inhibits 3CL^(pro)and PL^(pro)of SARS-CoV-2 virus and regulates immune response and inflammation of host cells for the treatment of COVID-19

It is an urgent demand worldwide to control the coronavirus disease 2019(COVID-19)pandemic caused by the severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)virus.The 3-chymotrypsin-like protease(3CL^(pro))and papain-like protease(PL^(pro))are key targets to discover SARS-CoV-2 inhibitors.After screening 12 Chinese herbal medicines and 125 compounds from licorice,we found that a popular natural product schaftoside inhibited 3CL^(pro)and PL^(pro)with IC_(50)values of 1.73±0.22 and 3.91±0.19μmol/L,respectively,and inhibited SARS CoV-2 virus in Vero E6 cells with EC_(50)of 11.83±3. 23μmol/L. Hydrogen-deuterium exchange mass spectrometry analysis, quantum mechanics/molecular mechanics calculations, together with site-directed mutagenesis indicated the antiviral activities of schaftoside were related with non-covalent interactions with H41, G143 and R188 of3CLpro, and K157, E167 and A246 of PLpro. Moreover, proteomics analysis and cytokine assay revealed that schaftoside also regulated immune response and inflammation of the host cells. The antiinflammatory activities of schaftoside were confirmed on lipopolysaccharide-induced acute lung injury mice. Schaftoside showed good safety and pharmacokinetic property, and could be a promising drug candidate for the prevention and treatment of COVID-19.

Differentiation of Belamcandae Rhizoma and Iridis Tectori Rhizoma by Thin-Layer Chromatography and High-Performance Liquid Chromatography

Objective:Belamcandae Rhizoma and Iridis Tectori Rhizoma are easily confused with each other.The main objective of this study is to distinguish them using chemical analysis.Materials and Methods:Thin-layer chromatography(TLC)and high-performance liquid chromatography(HPLC)fingerprint methods were established to compare the chemical profile,while HPLC quantitation was used to determine the contents of three isoflavones in thirty batches of Belamcandae Rhizoma and Iridis Tectori Rhizoma samples.Results:The two herbs could be distinguished by TLC using acetic acid-n hexane-ethyl acetate(1:90:80 v/v/v)as the mobile phase,according to the fluorescent band under 366 nm at R_(f) 0.2.In total,12 compounds were identified in the 24-min HPLC fingerprint.The similarity coefficient between the two herbs was 0.54±0.01.Mangiferin(1),tectoridin(2),iridin(3),irigenin(5),irisflorentin(6),and iristectorin A(9)were the main peaks in Belamcandae Rhizoma,while tectoridin(2)and tectorigenin(4)were the major peaks in Iridis Tectori Rhizoma.The contents of 2 in Iridis Tectori Rhizoma(2.50±0.20%)were 8.93 times higher than that of Belamcandae Rhizoma(0.28±0.08%),while the ones of 5 and 6 were slightly lower in Iridis Tectori Rhizoma.Conclusions:The study established fast and effective methods to distinguish Belamcandae Rhizoma from Iridis Tectori Rhizoma.