[Theoretical innovation of component-based Chinese medicine and its exemplary practice: the study on creating Guanxinning Tablets].

Upholding the wisdom of traditional Chinese medicine that the therapeutic principle, method, formula and medicine are coherent with each other, we propose the technical methodology for intelligent creation of component-based Chinese medicine by integrating multidisciplinary knowledge such as artificial intelligence, pharmaceutical informatics, system pharmacology and phytochemistry. Taking the creation of Guanxinning Tablets as an example, we expound the technical principle for creating component-based Chinese medicine and briefly describe the design method for optimizing the entity of Chinese medicine efficacy by rational combination of active components. Our research sought to "clarify and explain" the mechanism of its clinical treatment action through multi-modal and multi-scale systematic pharmacology studies. This work emphatically demonstrates the pilot workshop and engineering validation platform based on the intelligent simulation of whole production process, and outlines the design principles of the intelligent production line for innovative Chinese medicine. The results of industrial research show that the ourself established method for evaluating the process quality controllability and intelligent production line can be applied to manufacturing Guanxining Tablets with high quality. Through the innovative research of multidisciplinary cross-border integration, the present work explored a new way for the creation of modern Chinese medicine.

[Mechanism of Guanxinning against cerebral ischemia-reperfusion injury in mice based on transcriptomic analysis].

Guanxinning, a modern Chinese medicine preparation composed of Salviae Miltiorrhizae Radix et Rhizoma and Chuanxiong Rhizoma, has the activities of activating blood circulation, resolving blood stasis, dredging vessels, and nourishing the heart. Clinical studies have demonstrated that Guanxinning has therapeutic effect on ischemic stroke, while the specific mechanism remains to be clarified. In this study, the potential mechanism of Guanxinning against cerebral ischemia-reperfusion injury in mice was explored and then verified in vitro. The mouse model of cerebral ischemia-reperfusion injury was established with middle cerebral artery embolization(MCAO) method. The pharmacological effects of Guanxinning on the model mice were investigated based on neurological function score, cerebral infarction area, pathological morphology, neuron injury, and apoptosis. The results showed that Guanxinning lowered neurological functional score, reduced cerebral infarction area, and ameliorated the histopathological morphology, neuronal damage, and apoptosis in the model mice. RNA samples were extracted from brain tissues and subjected to RNA sequencing(RNA-seq). The differentially expressed genes(DEGs) were screened with the thresholds of ■. GO function enrichment analysis and KEGG pathway enrichment analysis were performed for the 297 common DEGs, which indicated that Guanxinning may regulate the inflammatory response, oxidative stress response, energy metabolism, and apoptosis to treat cerebral ischemia-reperfusion injury in mice. Guanxinning exerted protective effect through inhibiting inflammation and reducing oxidative stress in hypoxia/reoxygenation injured SH-SY5 Y cells. Furthermore, Western blot indicated that Guanxinning down-regulated the protein levels of p-NF-κB p65 and p-p38 MAPK and up-regulated those of PPARγ and PGC-1α. The findings suggested that Guanxinning may inhibit inflammation and reduce oxidative stress by suppressing TNF signaling pathway and activating PPAR signaling pathway, thereby exerting the therapeutic effect on cerebral ischemia-reperfusion injury in mice. This study preliminarily reveals the mechanism of Guanxinning against cerebral ischemia-reperfusion injury and provides a basis for clinical application of Guanxinning.

[Effects of Angong Niuhuang Pills against heart failure based on cross-scale polypharmacological study].

Angong Niuhuang Pills(AGNHP) are effective in clearing heat, removing the toxin, and eliminating phlegm for resuscitation. Clinically, it is widely used to treat various diseases such as febrile convulsion due to heat attacking pericardium, but its therapeutic effects on heart failure(HF) have not been well recognized. In this study, the profiles of differential metabolites regulated by AGNHP were identified by ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS). The underlying mechanism of AGNHP against HF was illustrated based on the integrated analysis of pharmacological data and metabolic molecular network. The HF model was induced by isoproterenol in mice. After oral administration of AGNHP for one week, cardiac functions in HF mice were evaluated by echocardiography, and serum samples of mice were collected for metabolomics analysis. Eight differential metabolites of AGNHP against HF were screened out through partial least square discriminant analysis(PLS-DA) and input into MetaboAnalyst for the analysis of metabolic pathways. Moreover, the critical metabolic pathways regulated by AGNHP were enriched according to the potential targets of major compounds in AGNHP. After AGNHP treatment, the recovered index of relative content of some metabolites underwent cross-scale fusion analysis with therapeutic efficacy data, followed by "compound-reaction-enzyme-gene" network analysis. It is inferred that the anti-HF effects of AGNHP may be attributed to the metabolism of arachidonic acid, amino acid, glycerophospholipid, and linoleic acid. The cross-scale polypharmacological analysis method developed in this study provides a new method to interpret scientific principles of AGNHP against HF with modern technologies.

[Methodology and applications for multimodal identification of active constituents of traditional Chinese medicine].

The discovery of active constituents of traditional Chinese medicine(TCM) faces multiple challenges, such as limited approaches to evaluate poly-pharmacological effects, and the lack of systematic methods to identify active constituents. Aimed at these bottleneck problems in the field, the present study intensively discussed the key scientific problems in the identification of active constituents of TCM, based on scientific methodologies including systematology, information theory, and synergetics. A comprehensive strategy is herein proposed to investigate the correlations between the chemical composition and biological activities of TCM, from macro-, meso-, and micro-scales. Moreover, in this study, we systematically proposed the methodology of the multimodal identification of TCM active constituents, and thoroughly constructed its core technologies. Its technical framework is suggested to be assessed by multimodal information acquisition, centered on multisource information fusion, and focused on interaction evaluation. Furthermore, the core technologies for the multimodal identification of active constituents of TCM were developed in this study, which is according to the characteristics of the exchanges of between TCM and biological organisms, in the aspects of material, energy and information. Finally, two examples of the application of the proposed method were briefly introduced. The proposed methodology provides a novel way to solve the bottlenecks in the study of active constituents of TCM, and lays the foundation for the multimodal study of TCM.

Dendrimer-Au Nanoparticle Network Covered Alumina Membrane for Ion Rectification and Enhanced Bioanalysis.

Ion transport in an artificial asymmetric nanoporous membrane, which is similar to biological ion channels, can be used for biosensing. Here, a dendrimer-Au nanoparticle network (DAN) is in situ assembled on a nanoporous anodic aluminum oxide (AAO) surface, forming a DAN/AAO hybrid membrane. Benefiting from the high surface area and anion selectivity of DAN, the prepared DAN/AAO hybrid presents selective ion transport. Under a bias potential, a diode-like current-potential (I-V) response is observed. The obtained ionic current rectification (ICR) property can be tuned by the ion valence and pH value of the electrolyte. The rectified ionic current endows the as-prepared DAN/AAO hybrid with the ability of enhanced bioanalysis. Sensitive capture and detection of circulating tumor cells (CTCs) with a detection limit of 80 cells mL-1 as well as excellent reusability can be achieved.

[Intelligent and lean manufacturing for Chinese medicine:theory and its applications].

To cope with the " six major scientific problems" and the " five major technical challenges" of intelligent manufacturing and lean production of Chinese medicine( CM),we systematically proposed strategies,methods and the engineering theory of intelligent and lean manufacturing for CM by integrating the holistic view of traditional Chinese medicine and the concepts inspired from international advanced pharmaceutical technology. Moreover,the translational research of the theory and methods was successfully applied to six CMs such as Xuesaitong Injection. Several intelligent production lines were designed and built on the basis of the theory and methods,which greatly accelerated the digitalization,networking,and intelligence manufacture for CM. As a conclusion,the theory and applications provide technical demonstration for technical upgrading and high-quality development of CM industry.

ω-3 Polyunsaturated Fatty Acid Postconditioning Protects the Isolated Perfused Rat Heart from Ischemia-Reperfusion Injury.

AIMS: This study aimed to evaluate the cardioprotective effects of ω-3 polyunsaturated fatty acids (PUFAs) postconditioning against ischemia-reperfusion (I/R) injury. METHODS: Sixty Sprague-Dawley rats were randomly divided into 4 groups (n = 15 for each) and used to generate the Langendorff isolated perfused rat heart model. The sham group received a continuous perfusion of 150 min. The remaining three I/R-treated groups sequentially received a 30-min perfusion, a 30-min cardioplegia, and a 90-min reperfusion. The I/R-ischemic preconditioning (IP) group additionally received three cycles of 20-s reperfusion and 20-s coronary reocclusion preceded the 90 min of reperfusion. The I/R-ω group were perfused with ω-3 PUFAs for 15 min before the 90 min of reperfusion. The myocardial infarct size, the degree of mitochondrial damage, the antioxidant capacity of the myocardium, and the cardiac functions during reperfusion were compared among groups. RESULTS: Compared with the I/R group, the I/R-ω group had significantly reduced myocardial infarct size, reduced levels of lactate dehydrogenase and malondialdehyde, elevated superoxide dismutase level, and elevated rising (+dp/dtmax) and descending (-dp/dtmax) rate of left ventricular pressure. The I/R-ω group had a significantly lower rate of mitochondrial damage in myocardial tissue compared with the I/R and I/R-IP groups. CONCLUSION: ω-3 PUFA postconditioning possesses good cardioprotective effects and may be developed into a therapeutic strategy for myocardial I/R injury.

Asymmetric Nanochannel-Ionchannel Hybrid for Ultrasensitive and Label-Free Detection of Copper Ions in Blood.

Nanochannel/nanopre based analysis methods have attracted increasing interest in recent years due to their exquisite ability to reveal changes in molecular volume. In this work, a highly asymmetric nanochannel-ionchannel hybrid coupled with an electrochemical technique was developed for copper ion (Cu2+) detection. Polyglutamic acid (PGA) was modified in a nanochannel array of porous anodic alumina (PAA). When different concentrations of Cu2+ were introduced into the nanochannel-ionchannel hybrid in a neutral environment, a Cu2+-PGA chelation reaction occurs, resulting in varied current-potential (I-V) properties of the nanochannel-ionchannel hybrid. When PAA was immersed in a low pH solution, the Cu2+-PGA complex dissociated. On the basis of the change in ionic current, a label-free assay for Cu2+ was achieved along with the ability to regenerate and reuse the constructed platform. Because of the unique mass transfer property of the nanochannel-ionchannel hybrid combined with the highly amplified ionic current magnitude of the nanochannel array, significantly increased assay sensitivity was achieved, as expected. To evaluate the applicability of the present methodology for detecting Cu2+ in a real sample, the Cu2+ content in real blood samples was analyzed. The results demonstrate that the present method shows excellent selectivity with high sensitivity toward Cu2+ detection in real blood samples.

Autophagy Induced by Areca Nut Extract Contributes to Decreasing Cisplatin Toxicity in Oral Squamous Cell Carcinoma Cells: Roles of Reactive Oxygen Species/AMPK Signaling.

Chewing areca nut is closely associated with oral squamous cell carcinoma (OSCC). The current study aimed to investigate potential associations between areca nut extract (ANE) and cisplatin toxicity in OSCC cells. OSCC cells (Cal-27 and Scc-9) viability and apoptosis were analyzed after treatment with ANE and/or cisplatin. The expressions of proteins associated with autophagy and the AMP-activated protein kinase (AMPK) signaling network were evaluated. We revealed that advanced OSCC patients with areca nut chewing habits presented higher LC3 expression and poorer prognosis. Reactive oxygen species (ROS)-mediated autophagy was induced after pro-longed treatment of ANE (six days, 3 µg). Cisplatin toxicity (IC50, 48 h) was decreased in OSCC cells after ANE treatment (six days, 3 µg). Cisplatin toxicity could be enhanced by reversed autophagy by pretreatment of 3-methyladenine (3-MA), N-acetyl-l-cysteine (NAC), or Compound C. Cleaved-Poly-(ADP-ribose) polymerase (cl-PARP) and cleaved-caspase 3 (cl-caspase 3) were downregulated in ANE-treated OSCC cells in the presence of cisplatin, which was also reversed by NAC and Compound C. Collectively, ANE could decrease cisplatin toxicity of OSCC by inducing autophagy, which involves the ROS and AMPK/mTOR signaling pathway.

[Establishment pharmacological research platform for "Concurrent treatment of the brain and heart" and its application on dissecting mechanism of Danhong injection].

The Concurrent treatment of the brain and heart (CTBH) theory is proposed based on traditional Chinese medical theory and clinical practice. In this study, a framework for the pharmacological research platform was established to investigate the principles of concurrent treatment of the brain and heart. The platform for CTBH includes several key techniques for network modeling, discovery of active substances, dissecting mechanism of action and investigation of pharmacokinetic property of TCM. Taking network modeling of CTBH as an example, using database search, literature mining, network construction and module analysis, the that network modules closely associated with the pathological progress of cardiovascular and cerebrovascular diseases were identified, while further functional enrichment analysis of these modules indicated that the key biological processes included oxidative stress, metabolism and inflammation. GSK3B, NOTCH1, CDK4 were identified as key nodes in these network modules. The above-mentioned platform was applied to construct component-biomolecules network of Danhong injection for the identification of common targets and pathways. Among them, GSK3B had the highest correlation with the composition of Danhong injection in the network, and the biological function of whose cluster was related to cell oxidative stress. Based upon results of network analysis, validation experiments suggested that Danhong injection significantly improved the survival rate of oxidative injured myocardial cells and nerve cells, and the protective effect was related to the increase of phosphorylated GSK3ß protein expression. Moreover, extracts of Salviae Miltiorrhizae Radix et Rhizoma and Carthami Flos exerted the synergisticcytoprotective effect. The results indicated that the mechanism of treatment of cardiovascular and cerebrovascular diseases of Danhong injection could be studied through network modeling and other methods. In summary, the proposed pharmacological platform provided a feasible way for revealing the mechanism of CTBH by using modern scientific methods.

[Screening and identification of DPP-4 inhibitors from Xiaokean formula by a fluorescent probe].

Fluorescent bio-probes have attracted increasing attentions in studies for screening bioactive compounds from traditional Chinese medicines. In this study, a new-type fluorescent probe with the function of aggregation-induced emission (AIE) was used to screen dipeptidyl peptidase-4 (DPP-4) inhibitor from Xiaokean formula, which has been clinically used for the treatment of type 2 diabetes mellitus. Potential DPP-4 inhibitors were screened by the fluorescent probe, with diprotin A as the positive control; totally 43 components were isolated from Xiaokean formula by systematic separation. The results showed that 13 components can exert inhibitory effects on DPP-4 activity; 16 compounds were further identified by liquid chromatography-mass spectrometry (LC-MS) from those active components. The inhibitory effects of 14 compounds were further verified, while five of them showed significant inhibition against DPP-4. Salvianolicacid C, ginsenoside Rg5 and timosaponin AI inhibited DPP-4 activity at the concentration of 5-50 µmol•L⁻¹ in a dose-dependent manner. Thus, our study provided a successful example for screening bioactive compounds from traditional Chinese medicines by using a novelfluorescent probe.

[Study on main pharmacodynamic effects for Schisandra lignans based upon network pharmacology].

In this study, we focused on the study of pharmacodynamic effects for 6 major bioactive lignans of Schisandra chinensis, namely deoxyschizandrin, schisandrin B, schisandrin C, schisandrin, schizandrol B and schisantherin A. A compound-gene-pathway network, which contained 124 related genes and 88 pathways, was constructed by collecting drug genes through mining relevant literatures and network pharmacology analysis. Based on the network analysis, 32 pathways and 80 related genes were associated with inflammation, which implied that anti-inflammatory might be the major pharmacodynamic effects of these compounds. All lignans except schizandrol B reduced LPS-induced NO production in RAW264.7 cells, which validated the anti-inflammatory hypothesis generated from network analysis. Furthermore, we investigated the effects of deoxyschizandrin, schisandrin C, schisandrin and schisantherin A on the secretion of inflammatory cytokines TNF-α, IL-1ß, IL-6, PGE2 and protein expressions of iNOS, COX-2. As a result, deoxyschizandrin showed the strongest anti-inflammatory activity with inhibitory effect on all 4 inflammatory cytokines secretions and protein expressions of iNOS, COX-2. This study provided evidences for systematic exploration on the pharmacolgical actions and mechanisms of schisandra.

[Study of screening nephroprotective bioactive substances based on triple-color fluorescence probes in Carthami flos].

In this study, an approach based on triple-color fluorescence probes was developed for screening potential nephro-protective bioactive substances. Three fluorescent probes (i. e. FDA, MTR and Hoechst 33342) were used to label HK-2 cells injured by doxorubicin hydrochloride, and cellular fluorescence images were subsequently acquired and analyzed by a cellular-fluorescence image microscopy platform. The established method was applied to screening 53 components of Carthami Flos, and three components C17, C18 and C19 were found to exhibit nephroprotective effects against doxorubicin hydrochloride induced injury on HK-2 cells. Eight compounds (i. e. hydroxysafflor yellow A, 6-hydroxykaempferol-3-O-rutinoside-6-O-glucoside, 6-hydroxykaempferol-3,6-di-O-gluco-side or 6-hydroxykaempferol-6, 7-di-O-glucoside, 6-hydroxykaempferol-3-O-rutinoside, 6-hydroxykaempferol-3-O-glucoside or 6-hydroxykaempferol-7-O-glucoside, rutin, isoquercetin, and kaempferol-3-O-rutinoside) in components C17, C18 and C19 were preliminarily identified by liquid chromatography-mass spectrometry (LC-MS). Isoquercetin, rutin, kaempferol-3-O-rutinoside, and hydroxysafflor yellow A were confirmed by comparing with reference substances, Further study indicated that these four compounds had moderate nephroprotective effects, while isoquercetin showed a significant nephroprotective effect in a dose-dependent manner. These results suggest that isoquercetin, rutin, kaempferol-3-O-rutinoside and hydroxysafflor yellow A might be the nephroprotective bioactive substances in Carthami Flos.

[Network pharmacology study of mechanism on xuesaitong injection against retinal vein occlusion].

Retinal vein occlusion (RVO) is a common clinical disease causing vision loss. Risk factors such as diabetes, atherosclerosis are closely associated with RVO. Xuesaitong injection is used extensively in clinical treatment of RVO, however the mechanism is still unclear. In this study, we investigated the protective effect of Xuesaitong injection on RVO rat model. Using a compound-target network of Xuesaitong on anti-RVO constructed by literature mining, we aim to elucidate the multi-compound, multi-target effect of Xuesaitong injection. Fifteen potential targets of Xuesaitong injection associated with inflammation, angiogenesis, apoptosis, and coagulation were identified in this study. VEGF, IL-1beta and IL-6, three important targets in the compound-target network were further experimentally validated. This study provided experimental evidence for Xuesaitong injection being effective in treating RVO and a network view on its anti-RVO mode of action through a multi-compound and multiple-target mechanism.

[Strategies and key technologies for risk control and management in quality of Chinese medicine injections based on integrated pharmacology].

This paper focuses on the quality risk control and management of Chinese medicine (CM) injections. The most important technological requirements are analyzed, and a strategy for integrated pharmacology to study CM mechanism is proposed. A key technology system for quality risk control and management was further constructed. The strategy and technology system was finally applied to Shengmai injection for quality risk control and management.

[Study on rapid screening and identifying hepatotoxic compounds in toosendan fructus].

Fluorescein diacetate-labeled HepG2 cells model and flouresence automatic microscopy screening assay were used for fast screening 23 components from Toosendan Fructus, in which 5 components showed significant toxicity on HepG2 cells. The 10 compounds in the 2 components were tentatively identified with LC-MS(n), and 3 of them (meliasenin B, trichilinin D and 1-O-tigloy-1-O-debenzoylohchinal) were prepared and identified. Further experiments showed that the 3 compounds displayed dose-dependent toxicity on HepG2 cells, suggesting that these compounds in Toosendan Fructus may cause hepatotoxicity.

[An approach to screen nephrotoxic components in traditional Chinese medicine based on fluorescent probe and HK-2 cells and its application].

Using fluorescein diacetate labeled HK-2 cells, a fast method for screening nephrotoxic components in traditional Chinese medicines (TCMs) was proposed in this study. The methodology validation showed that the linearity, stability and accuracy of the proposed method were suitable for screening nephrotoxic components in vitro . This method was further applied on screen 352 components from 32 Chinese Pharmacopoeia-indexed toxic TCMs. The results indicated that 31 components from 14 toxic TCMs, including Badou, had significant toxicity on HK-2 cells, which suggested these components may cause nephrotoxicity. The components from the other 18 toxic TCMs had no significant toxicity on HK-2 cells.

[Development of a rapid screening method for discovering neuroprotective components from traditional Chinese medicine].

This study developed a method for rapid screening neuroprotective compounds with FDA (fluorescein diacetate) labeled SH-SYSY cells, which was injuried by Glu (L-glutamic acid). The cell viability was determined by fluorescence automatic mi-croscopy screening system. Then, neuroprotective components from Gegen Qinlian decoction were screened by the proposed method. The results demonstrated that 4 chemical components, C15, D06, D07 and E05 from Gegen Qinlian decoction, showed significantly neuroprotective effects. Furthermore, 8 compounds (i. e. daidzin, 3'-methoxydaidzin, liquiritin apioside, 6-C-L-alpha-arabinopyranosyl-8-C-beta-D-glucopyranosyl chrysin, isoliquiritin apioside, baicalin, oroxylin-A-7-O-fP-D-glucuronide and wogonoside) were identified from these active components by LC-Q/TOF-MS technology (liquid chromatography quadrupole time of flight tandem mass spectrometry). These 8 compounds may be the potential neuroprotective substances in Gegen Qinlian decoction.

[Study on screening cardioprotective components from Zhishi Xiebai Guizhi Tang].

Zhishi Xiebai Guizhi Tang (ZSXBGZT), a famous traditional Chinese formula from Synopsis of Golden Chamber, composing of five herbal medicines, has been routinely used for the therapy of cardiovascular diseases in clinical. However, the active components of ZSXBGZT have not been investigated thoroughly. A reliable cell model was established by H2O2-induced injury in neonatal rat cardiomyocytes (NRCMs), with fluorescein diacetate (FDA) labelling the live cells, to screen cardioprotective substances. Seven components were found to have obvious cardioprotective effects through screening ZSXBGZT, Vc as the positive control. Eleven compounds were identified by liquid chromatography-mass spectrometry (LC-MS) from the active components C18, D14, D15, D16 and E09, and 6 of them were investigated further, which showed that hesperidin, neohesperidin and eriodictyol could protect NRCMs from H2O2-induced injury. In particular, eriodictyol showed significant cardioprotective effects in a dose-response relationship.

[Screening and identifying hepatotoxic components in Aucklandiae Radix with GC-MS].

OBJECTIVE: To screen the potential hepatotoxic components in Chinese herb medicine Aucklandiae Radix. METHODS: The potential hepatotoxic components were screened using HepG2 cells labeled with fluorescein diacetate from 25 fractions of Aucklandiae Radix, in which the hepatotoxic compounds were further identified with GC-MS. RESULTS: Ten potential hepatotoxic fractions were screened. The identification results by GC-MS indicated that the main compounds in C09 were dehydrocostuslactone, santamarine (or magnolialide) and reynosin, and in C11 were α-costol and elemol. CONCLUSION: Dehydrocostuslactone, santamarine (or magnolialide), reynosin, α-costol and elemol are potential hepatotoxic compounds in Aucklandiae Radix.