Arenaviruses are enveloped RNA viruses, and eight members in this family are known to cause human hemorrhagic fever. Treatments for the viral hemorrhagic fever (VHF) by arenaviruses are very limited. We have identified the first flavone, tangeretin, with broad-spectrum inhibitory activities on VHF-arenaviruses infection by blocking viral entry. In this study, we evaluated thirty-four tangeretin analogues and found 3, 5, 6, 7, 4'-pentamethoxyflavone as a Lassa virus entry inhibitor, with EC₅₀ of 5.2 μmol·L^-1, by blocking the viral fusion pro cess. The compound 3, 5, 6, 7, 4'-pentamethoxyflavone is effective on all known VHF-arenaviruses, with EC₅₀ range of 0.84-10.2 μmol·L^-1. These results suggest that 3, 5, 6, 7, 4'-pentamethoxyflavone is able to serve as a start point for discovery of arenavirus entry inhibitors from flavone natural products.

Among various technologies used in drug design and discovery, deep learning is still in its infancy. Recently, deep learning approaches have been rapidly developed and applied to address various problems in drug discovery, including generation of virtual compound library, prediction of compound activity, metabolism and toxicity, and prediction of organic synthesis routes. Compared with the traditional machine learning methods, the prediction power of deep learning did not show significant improvement. However, proactively learning and automatically feature extraction bring advantages for deep learning approaches. Compared to first principle-based computational chemistry methods, deep learning can not be generalized because it depends on large-scale and highquality annotated data sets. But its molecular representation with single-atom atomic environment vectors could be useful for computational chemists. As an emerging technology, deep learning, especially the unsupervised learning method that does not rely on large datasets with labels, is gradually improving. It is expected that someday deep learning method will become practical for drug discovery.

Snake venom has special pharmacological activities and contains a array of small polypeptides that can antagonize integrins, therefore called disintegrins. Disintegrins can block integrin-dependent platelet aggrega tion, tumor growth, and tumor metastasis. A disintegrin fraction was isolated and purified from the venom of snake Gloydius brevicaudus (GBV). Its physical and chemical properties were characterized, and its biological activities were investigated. The crude venom of GBV were isolated by Superdex 75 gel filtration chromatography. The antiplatelet aggregation activity of the fractions was screened by the Born method. The fraction that shown anti-platelet activity was further purified with Sephadex G-25 gel filtration, DEAE Sepharose Fast Flow ion exchange chroma tography, and Lichrospher C18 reversed-phase chromatography respectively. The purity of the active component was analyzed with SDS-PAGE (Tris-Tricine system) and high-performance liquid-phase chromatography (HPLC), with protein concentration determined by the Bradford method. The molecular weight was evaluated by the gel imaging method and mass spectrometry, and the isoelectric point was measured by disc isoelectric focusing electro phoresis. The protease activity was measured with the Rick method. The phospholipase A activity was determined by the automatic potentiometric titration method. Amino acid sequencing results were subjected to homology comparison using the BLAST program. Seven fractions (Ⅰ-Ⅶ) were isolated from GBV by gel filtration chroma tography on Superdex 75 column. The fraction Ⅳ inhibited the platelet aggregation induced by ADP with molecular weight lower than 10 000 Da, suggesting a disintegrin component. A disintegrin named GBV-Ⅳ 4 was purified from the fraction by Sephadex G-25 gel filtration, DEAE Sepharose Fast Flow ion-exchange and Lichrospher C18 reverse chromatography. It was homogeneous shown as a single band on SDS-polyacrylmide gel electrophoresis (SDS-PAGE, Tris-Tricine system) with molecular weight 8 746 Da as calculated by Image Master VDS system. The isoelectric point of GBV-Ⅳ4 was 6.3 by disc polyacrylamide gel electrophoresis. GBV-Ⅳ4 exhibited no detectable phospholipase A₂ (PLA₂) activity with the pH-stat technique or proteinase activity according to the method of Rick. GBV-Ⅳ4 is composed of 70 amino acids with RGD (Arg-Gly-Asp) active region and a molecular weight of 7 442 Dalton as assayed by Mass Spectrography. Characterization of GBV-Ⅳ4 is consistent with metachain disintegrin (70 amino acid sequence, six pairs of disulfide bond). Retrieved by Genbank, GBV-Ⅳ4 has high homology with other disintegrins. We concluded that GBV-Ⅳ 4 is a novel disintegrin contained RGD. GBV-Ⅳ4 showed dose-dependent inhibition of ADP-or thrombin-induced platelet aggregation with IC₅₀ 0.339 or 0.577 μg·mL^-1 respectively. In conclusion, a new disintegrin derived from the GBV snake venom and named GBV-Ⅳ4 containing RGD tripeptide sequence could inhibit platelet aggregation.

Nitidine chloride (NC) is a compound with prominent anti-tumor activity. To determine potential cardiotoxicity of NC, this study was designed to investigate the distribution of NC in rat heart and the underlying mechanism. The animal studies were approved by Institutional Animal Care and Use Committee of Zhejiang University Medical Center (2015-380-01) and complied with the standards of animal welfare in China. At 0.25, 0.5 and 2 h after a single intravenous injection (iv) of 5 mg·kg^-1 NC, the concentrations of NC in rat heart were 47.7, 71.1 and 63.2 μg·g^-1 respectively, which were 576, 1 352 and 1 212 folds of that in plasma. This study also revealed that the NC concentration in heart was 458.5 μg·g^-1 (7 336 folds of that in plasma) at 2 h after the last dose in rats, after daily iv administration of NC at 5 mg·kg^-1·day^-1 for successive 20 days. Further studies showed that the accumulations of NC in MDCK-hOCT1 and MDCK-hOCT3 cells were 16.1 and 4.99 folds higher than that of the mock cells, respectively. There is no significant difference between the accumulations of NC in MDCK cells transfected with hOCTN1, hOCTN2 or hPMAT and the mock cells. Additionally, quinidine, L-tetrahydropalmatine and Decynium 22, the inhibitors of OCTs, clearly reduced the accumulations of NC in primary cardiomyocytes and cardiac fibroblasts from neonatal rats. MTT assay showed that the LC₅₀ of NC on cardiomyocytes and cardiac fibroblasts were 10.9 and 10.4 μmol·L^-1, respectively. Moreover, treatment of the primary cardiomyocytes and cardiac fibroblasts with NC (1~15 μmol·L^-1) for 48 h resulted in significantly increased LDH enzyme leakage. These results indicated that NC can be highly accumulated in the heart, and accumulation is mediated by OCT1 and OCT3, but not by OCTN1, OCTN2 and PMAT. The accumulated NC has potential cytotoxicity as shown in the results from primary cardiomyocytes and cardiac fibroblasts.

Based on special scientific facts demonstrated in traditional Chinese medicine (TCM) complex system, this paper proposes a hypothesis of "one output multi-source", discussing the concept, features, structures and the scope. The law of interaction between the integrity and multiple components of TCM complex system was examined. Feasibility, technical methods and evidence supporting the hypothesis have been presented here. We present a basic model of the hypothesis, i.e. artificial neural network (ANN) model. This hypothesis promotes a deeper modern scientific understanding towards the TCM complex system and advancement in research of the material basis. TCM compatibility and quality control will serve as the theoretical foundation for guiding the research on drug combination including chemical, biological and herbal medicines.

Tuberculosis (TB) is a serious infectious disease caused by Mycobacterium. tuberculosis. In recent years, with the emergence of drug-resistant forms, the development of new anti-tuberculosis drugs is urgently needed. In this study, we used Mycobacterium marinum (M. marinum), which is highly similar to M. tuberculosis, to establish a M. marinum infected-zebrafish model and quantitative PCR (qPCR) method for bacterial count analysis. The results showed that injecting M. marinum into the yolk sac is an efficient and convenient way to infect zebrafish embryos. By counting the survival rate of infected zebrafish and the number of bacteria in zebrafish by ZiehlNeelsen staining, we analyzed the efficacy of isoniazid and rifampicin as anti-tuberculosis drugs and the synergistic effect of drugs. The results suggested that three evaluation methods exhibit good consistency. This study demon strated that zebrafish-M. marinum infection model combined with qPCR analysis is a simple and efficient method for in vivo screening and evaluation of anti-tuberculosis drugs. Animal experiments were carried out in accordance with the provisions for animal ethics in the Regulations on Laboratory Animals of Institute of Medicinal Biotech nology, Chinese Academy of Medical Sciences.

The compatibility of traditional Chinese medicines (TCM) includes the compatibility of single herb, effective parts and the effective ingredients of TCM herbs. Compared with the compatibility of Chinese herbal medicine, the compatibility between effective parts and effective ingredients of TCM has become a breakthrough point in the study of compatibility. It provides a new method for precise medication research of TCM, which is bound to become an important driving force in the process of precision medicine research. In this paper, we elabo rate the strategy of the compatibility and proportion in TCM components from five aspects:the compatibility mode of effective components in TCM, the screening method of compatibility of components in TCM, the selection of compatibility component dose, the design method of proportion test and the optimization research of compatibility and proportion.

Based on the concept of network pharmacology, the main nephroprotective components in Erzhi Pill reported in previous studies, were used to predict the targets through the PharmMapper method. Molecular docking was applied to screen for potential targets and biological information annotation databases (DAVID) was used to analyze the molecular function and biological process of the action targets. The Cytoscape software was used to construct the "ingredient-target-pathway" network of Erzhi Pill for renal injury treatment. TTD and GAD database were then applied to screen for the targets of renal disease for building "ingredient-core target" network. We found that 17 major active ingredients of Erzhi Pill regulated 32 targets (including ESR1, ESR2, GCK, MMP3) and affected 6 pathways, such as PI3K-Akt signaling pathway, estrogen signaling pathway and purine metabolism. This study reflected the nature of traditional Chinese medicine as multi-ingredients, multi-targets and multi-path ways, providing new clues for basic science research on the nephroprotective pharmacological mechanism of Erzhi Pill.

The purpose of this research is to investigate the effects of acacetin on serum lipid metabolism and atherosclerosis in mice and explore its molecular mechanism. HepG2 cells were treated with different concentrations of acacetin. The expression of LDL receptor (LDLR) and sterol-regulatory element binding protein-2 (SREBP-2) were detected by RT-qPCR and/or Western blot. C57BL/6J mice were given acacetin (50 mg·kg^-1) for 5 weeks by gavage. Serum total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C) and triglyceride (TG) were analyzed by an automatic biochemical analyzer. The expression of LDLR or SREBP-2 was detected by Western blot. After 12 weeks of intragastric administration of acacetin (30 mg·kg^-1) in apolipoprotein E knockout (ApoE KO) mice, the serum lipid levels were determined by an automatic biochemical analyzer. The lipid deposition in aortic plaque (en face) and aortic root plaque were stained with oil red O. The expression of LDLR and SREBP-2 were detected by RT-qPCR and/or Western blot. The intestinal content microflora was analyzed by 16S rDNA sequencing (All animal studies were approved by the Animal Experimentation Ethics Committee of Institute of Medicinal Biotechnology, CAMS & PUMC). In vitro results indicated that acacetin significantly up-regulated LDLR mRNA and protein levels, and stimulated LDLR transcription factor SREBP-2 protein expression. As indicated from in vivo studies, compared with control group, acacetin significantly decreased the serum levels of TC and LDL-C in C57BL/6J mice by 34% and 57% (P < 0.01), respectively. Furthermore, mechanic study showed that acacetin significantly increased the protein expression of hepatic LDLR and SREBP-2. Although the results of serum lipid profiles, hepatic LDLR/SREBP-2 expression and area of atherosclerotic lesions in aorta and aortic root in ApoE KO mice showed differences between acacetin and high-fat diet group, the differences did not reach statistical significance. Nevertheless, acacetin exhibited a profound influence on the composition of the intestinal microbiota as indicated by 16s rDNA sequencing analysis. In conclusion, these results demonstrated that acacetin can decrease the serum lipid levels in C57BL/6J mice through up-regulation of hepatic LDLR and SREBP-2, and alter gut microflora in high-fat diet fed Apo KO mice. This study suggests the possibility that acacetin has a potential role in inhibiting the progression of atherosclerosis.

The study was designed to synthesize a novel dendritic copolymer composed of polyamidoamine dendrimer G0 as the inner core and poly(L-glutamic acid) grafted low molecular weight polyethylenimine (PGLP) as surrounding arms for gene delivery vector. The molecular structure of PGLP was confirmed by ¹H NMR (proton nuclear magnetic resonance spectroscopy). The DNA combination capability of PGLP was examined by gel retarda tion electrophoresis. The particle sizes and zeta potentials of PGLP/pDNA complexes were determined by dynamic light scattering (DLS). The cytotoxicity of PGLP was evaluated by Cell Counting Kit-8 (CCK-8) and hemolysis assays, which was approved by Research Ethics Committee of the First Affiliated Hospital of Nanchang University. The in vitro transfection efficiency of PGLP was measured by a flow cytometry. The results of physicochemical properties suggested that PGLP could self-assemble with DNA to form complexes with average particle sizes of about 105-200 nm and zeta potentials of about +10-+28 mV, which could protect DNA from serum degradation. The results of biological properties suggested that PGLP showed more higher transfection efficiencies but lower cytotoxicity than PEI 25K or Lipofectamine 2000 in various cell lines (HEK 293T, HeLa, BEL 7402, RASMC). Importantly, it was found that PGLP/pDNA complexes at w/w=8 showed more strong serum-resistant capacity than PEI 25K/pDNA complexes. Therefore, PGLP is a promising candidate vector for gene delivery.