Chinese Medicine Library Research Articles

Computational screening and molecular docking of compounds from Traditional Chinese Medicine (TCM) by targeting DNA topoisomerase I to design potential anticancer drugs.

Each year thousands of people suffer across the globe due to higher cancer incidence and mortality rates. Additionally, the treatment option for cancer patients is also costly, and often cancer drugs suffer from lower efficacy with more side effects. The DNA topoisomerase can function as an established cancer target because Human Topoisomerase (Top1) regulates genetic transcription during the post-mitotic phase and plays a critical role in DNA supercoiling during replication and repair. Therefore, during drug therapy, blocking the Top1 may be crucial for inhibiting the proliferation of cancer cells. Here, the TCM (traditional Chinese medicine) compounds have been screened through the virtual screening. The Chinese medicine library's virtual screening process made it possible to narrow down the compound list to 29 compounds based on binding energy (-7.1 to -9.3Kcal/mol), while following Lipniski filtering, MM/PB (GB) SA filtering was used to screen the remaining 22 compounds and the top four compounds were chosen based on binding free energy. Here, the four compounds; CID-65752 (T2972: Rutaecarpine), CID-5271805 (T4S2126: Ginkgetin), CID-9817839 (T2S2335: Dehydroevodiamine) and CID-51106 (T3054: Daurisoline) had comparatively higher binding energy of -8.2, -8.5, -8.3 and -8.2 respectively during molecular docking than other compounds. Among these four compounds, no toxic profile of the two screened compounds; CID-5271805 and CID-9817839 was found in ADMET filtering. Moreover, the SASA (solvent accessible surface area), Rg (radius of gyrations), RMSD (root mean square deviation), and RMSF (root mean square fluctuation) profile of the drug-protein complex reveals the stability and rigidity of the compounds in molecular dynamics simulation study. However, these studies need to be validated in experimental approaches to develop more potent and effective cancer drugs.

In Silico Discovery of Potential Inhibitors Targeting the RNA Binding Loop of ADAR2 and 5-HT2CR from Traditional Chinese Natural Compounds.

Adenosine deaminase acting on RNA 2 (ADAR2) is an important enzyme involved in RNA editing processes, particularly in the conversion of adenosine to inosine in RNA molecules. Dysregulation of ADAR2 activity has been implicated in various diseases, including neurological disorders (including schizophrenia), inflammatory disorders, viral infections, and cancers. Therefore, targeting ADAR2 with small molecules presents a promising therapeutic strategy for modulating RNA editing and potentially treating associated pathologies. However, there are limited compounds that effectively inhibit ADAR2 reactions. This study therefore employed computational approaches to virtually screen natural compounds from the traditional Chinese medicine (TCM) library. The shortlisted compounds demonstrated a stronger binding affinity to the ADAR2 (<-9.5 kcal/mol) than the known inhibitor, 8-azanebularine (-6.8 kcal/mol). The topmost compounds were also observed to possess high binding affinity towards 5-HT2CR with binding energies ranging from -7.8 to -12.9 kcal/mol. Further subjecting the top ADAR2-ligand complexes to molecular dynamics simulations and molecular mechanics Poisson-Boltzmann surface area (MM/PBSA) calculations revealed that five potential hit compounds comprising ZINC000014637370, ZINC000085593577, ZINC000042890265, ZINC000039183320, and ZINC000101100339 had favorable binding free energies of -174.911, -137.369, -117.236, -67.023, and -64.913 kJ/mol, respectively, with the human ADAR2 protein. Residues Lys350, Cys377, Glu396, Cys451, Arg455, Ser486, Gln488, and Arg510 were also predicted to be crucial in ligand recognition and binding. This finding will provide valuable insights into the molecular interactions between ADAR2 and small molecules, aiding in the design of future ADAR2 inhibitors with potential therapeutic applications. The potential lead compounds were also profiled to have insignificant toxicities. A structural similarity search via DrugBank revealed that ZINC000039183320 and ZINC000014637370 were similar to naringin and naringenin, which are known adenosine deaminase (ADA) inhibitors. These potential novel ADAR2 inhibitors identified herein may be beneficial in treating several neurological disorders, cancers, viral infections, and inflammatory disorders caused by ADAR2 after experimental validation.

Metabolomic analysis of Thai Herbal Analgesic Formula based on ultra-high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry

Sahatsatara formula (STF), a Thai herbal medicine formula which has been commonly used as analgesic drugs to relieve musculoskeletal pain and numbness in Thai traditional medicine. The pharmacological studies of its ingredients have represented that have anti-inflammatory and antioxidant properties. However, the quality markers (Q-markers) for STF are still unknown and require further investigation. The primary goal of this study was to establish the chemical profile of STF though metabolomic analysis. Untargeted metabolomics were used to analyze global components and accurately qualify compounds. Multivariate analysis (MVA) was used to classify STF extract at three different concentrations and a quality control sample. Furthermore, samples' characteristics and identification-related markers were observed and compounds matched to the Traditional Chinese medicine library in UNIFI software. According to the results, chemical analysis revealed 63 compounds in positive mode and 33 compounds in negative mode within STF. Notably, 19 potential Q-markers were tentative identified in all three concentrations of STF, including alkaloids, terpenes, phenols, organic acids, disaccharides, fatty acids, glycosides, quinonoids, and steroids. The compounds exhibited pharmacological effects such as anti-inflammatory activity, anti-oxidant activity, and analgesic properties, which correlated to traditional properties of STF. Consequently, this study provides insights into the chemical profiles of the STF and identifies potential markers that can be utilized for qualitative and quantitative quality control of STF. Additionally, the findings can also be useful for further research into STF's anti-inflammatory properties through in vitro assays, as well as exploring its clinical efficacy to support evidence-based medicine for STF.

Glabridin inhibited the spread of polymyxin-resistant Enterobacterium carrying ICEMmoMP63.

The role of integrative and conjugative elements (ICEs) in antibiotic resistance in Morganella morganii is unknown. This study aimed to determine whether an ICE identified in the M. morganii genome contributed to the polymyxin resistance. Whole-genome sequencing was performed followed by bioinformatics analyses to identify ICEs and antibiotic resistance genes. Conjugation assays were performed to analyze the transferability of a discovered ICE. A drug transporter encoded on the ICE was heterogeneously expressed in Escherichia coli, minimum inhibitory concentrations of antibiotics were determined, and a traditional Chinese medicine library was screened for potential efflux pump inhibitors. An antibiotic resistance-conferring ICE, named ICEMmoMP63, was identified. ICEMmoMP63 was verified to be horizontally transferred among Enterobacteriaceae bacteria. G3577_03020 in ICEMmoMP63 was found to mediate multiple antibiotic resistances, especially polymyxin resistance. However, natural compound glabridin was demonstrated to inhibit polymyxin resistance. Our findings support the need for monitoring dissemination of ICEMmoMP63 in Enterobacteriaceae bacteria. Combined glabridin and polymyxin may have therapeutic potential for treating infections from multi-drug resistant bacteria carrying ICEMmoMP63.

Identification of chemical constituents in pomegranate seeds based on ultra-high-performance supercritical fluid chromatography coupled with quadrupole time-of-flight mass spectrometry.

To reveal the main chemical constituents of pomegranate seeds, a reliable and sensitive method based on ultra-high-performance supercritical fluid chromatography coupled with electrospray ionization and quadrupole time-of-flight mass spectrometry (MS) was developed. A time-dependent MSE data acquisition mode was applied to acquire the mass spectrometric data. The chemical constituents were identified by an automatic retrieval of a traditional Chinese medicine library and relevant literature. A total number of 59 compounds, including fatty acids, sterols, vitamins, cerebrosides, phospholipids, flavonoids, phenylpropanoids, and others, were tentatively identified. Their possible fragmentation pathways and characteristic ions were proposed and elucidated. The findings of this study, along with the developed methodology, could provide a reference for basic research on the pharmacodynamic substances of pomegranate seeds and shed light on their potential nutritional and therapeutic applications in the future.

Anti-hyperglycemic fraction from Alternanthera sessilis L. leaves gets elucidated following bioassay-guided isolation and mass spectrometry

The anecdotal use of Alternanthera sessilis L. as a relief for diabetes has been known in the Philippines for generations, and antidiabetic activity of similar varieties in other countries is likewise documented. However, the compounds responsible for this activity remain unclear. This study aims to isolate the anti-hyperglycemic fraction of local A. sessilis leaves and identify the compounds in this fraction. Methanol extract of A. sessilis leaves and its hexane, ethyl acetate (ASE), and water fractions were administered to alloxan-induced diabetic mice. ASE (250mg/kg) had the highest anti-hyperglycemic activity at 6-h post-treatment (25.81%±12.72%), with almost similar blood glucose reduction rate as metformin (30.13±3.75%, p=0.767). Repeated fractionation employing chromatographic separation techniques followed by in vivo anti-hyperglycemic assay yielded partially purified subfractions. A. sessilis ethyl acetate subfraction 4-2 (100mg/kg) displayed remarkable suppression of blood glucose rise in diabetic mice at 6-h post-treatment (26.45±3.75%, p<0.0001), with comparable activity with metformin (100mg/kg, 27.87±5.65%, p=0.652). Liquid chromatography/mass spectrometry showed eight distinct peaks, with four peaks annotated via the Traditional Chinese Medicine library and custom library for A. sessilis. Among these, luteolin, apigenin, ononin, and sophorabioside were identified as putative compounds responsible for the anti-hyperglycemic activity. This result provided basis for the reported anecdotal claims and potential utility of the local variety of A. sessilis leaves as sources of anti-hyperglycemic agents.

The traditional chinese medicine monomer Ailanthone improves the therapeutic efficacy of anti-PD-L1 in melanoma cells by targeting c-Jun

BackgroundC-Jun, a critical component of AP-1, exerts essential functions in various tumors, including melanoma, and is believed to be a druggable target for cancer therapy. Unfortunately, no effective c-Jun inhibitors are currently approved for clinical use. The advent of immune checkpoint inhibitor (ICI) has brought a paradigm shift in melanoma therapy, but more than half of patients fail to exhibit clinical responses. The exploration of new combination therapies has become the current pursuit of melanoma treatment strategy. This study aims to screen out Chinese herbal monomers that can target c-Jun, explore the combined effect of c–Jun inhibitor and ICI, and further clarify the related molecular mechanism. MethodsWe adopted a combinatorial screening strategy, including molecular docking, ligand-based online approaches and consensus quantitative structure-activity relationship (QSAR) model, to filter out c-Jun inhibitors from a traditional Chinese medicine (TCM) library. A mouse melanoma model was used to evaluate the therapeutic efficacy of monotherapy and combination therapy. Multicolor flow cytometry was employed to assess the tumor microenvironment (TME). Multiple in vitro assays were performed to verify down-streaming signaling pathway. CD4 + T-cell differentiation assay was applied to investigate Treg differentiation in vitro.ResultsAilanthone (AIL) was screened out as a c-Jun inhibitor, and inhibited melanoma cell growth by directly targeting c-Jun and promoting its degradation. Surprisingly, AIL also facilitated the therapeutic efficacy of anti-programmed death ligand-1 (PD-L1) in melanoma cells by reducing the infiltration of Tregs in TME. Additionally, AIL treatment inhibited c-Jun-induced PD-L1 expression and secretion. As a consequence, Treg differentiation was attenuated after treatment with AIL through the c-Jun/PD-L1 axis.ConclusionOur findings identified AIL as a novel c-Jun inhibitor, and revealed its previously unrecognized anti-melanoma effects and the vital role in regulating TME by Treg suppression, which provides a novel combination therapeutic strategy of c-Jun inhibition by AIL with ICI.Graphical AIL down-regulates c-Jun by reducing its stability, and inhibits the function of Tregs via AIL-c-Jun-PD-L1 pathway, ultimately suppressing melanoma progression and enhancing the efficacy of anti-PD-L1.

Ginkgolic acids inhibit SARS-CoV-2 and its variants by blocking the spike protein/ACE2 interplay

Targeting the interaction between the spike protein receptor binding domain (S-RBD) of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and angiotensin-converting enzyme 2 (ACE2) is a potential therapeutic strategy for treating coronavirus disease 2019 (COVID-19). However, we still lack small-molecule drug candidates for this target due to the missing knowledge in the hot spots for the protein-protein interaction. Here, we used NanoBiT technology to identify three Ginkgolic acids from an in-house traditional Chinese medicine (TCM) library, and they interfere with the S-RBD/ACE2 interplay. Our pseudovirus assay showed that one of the compounds, Ginkgolic acid C17:1 (GA171), significantly inhibits the entry of original SARS-CoV-2 and its variants into the ACE2-overexpressed HEK293T cells. We investigated and proposed the binding sites of GA171 on S-RBD by combining molecular docking and molecular dynamics simulations. Site-directed mutagenesis and surface plasmon resonance revealed that GA171 specifically binds to the pocket near R403 and Y505, critical residues of S-RBD for S-RBD interacting with ACE2. Thus, we provide structural insights into developing new small-molecule inhibitors and vaccines against the proposed S-RBD binding site.

Analysis of the chemical components of pomelo peels (Citrus grandis [L.] Osbeck) from different cultivars by using supercritical CO2 fluid extraction and ultra-high-performance liquid chromatography-tandem mass spectrometry.

Five pomelo cultivars (i.e., Citrus grandis cv. Shatianyou, Citrus grandis cv. Guanximiyou, Citrus grandis cv. Yuhuanyou, Citrus grandis cv. Duweiwendanyou and Citrus grandis cv. Liangpingyou) from different origins in China were selected to analyze their components by using supercritical CO2 fluid extraction coupled with ultra-high-performance liquid chromatography-tandem mass spectrometry. A total of 45 compounds were identified in the supercritical CO2 fluid extracts of the pomelo peels from the five cultivars. These compounds included eight flavonoids, 18 coumarins, four organic acids, three aldehydes, and 12 other compounds, which were identified using the obtained MS data and by comparison with commercial standards, orbitrap Chinese Traditional Medicine Library, and previous literature. Twenty-five of the identified compounds were detected for the first time in the pomelo peel extracts. Results suggested that the pomelo peels of C. grandis cv. Shatianyou contained the most natural chemical compositions. The pooled result may offer scientific evidence for further development and utilization of pomelo peels and a route for screening appropriate varieties for various demands.

Rapid Profiling of Chemical Constituents in Qingfei Paidu Granules Using High Performance Liquid Chromatography Coupled with Q Exactive Mass Spectrometry.

Qingfei Paidu (QFPD) granules have played a critical role during the Coronavirus Disease 2019 (COVID-19) in China. However, worldwide acceptance has been a problem because of the complex ingredients and unique theory of treatment. In this study, high-performance liquid chromatography (HPLC)-Q Exactive Orbitrap-mass spectrometry (MS) and the Orbitrap traditional Chinese medicine library (OTCML) were used to investigate the chemical constituents of QFPD granules. By comparing retention times, masses, isotope ion patterns, and MS2 profiles, 108 compounds were putatively identified using the OTCML combined with manual verification, including 12 alkaloids, 49 flavonoids, 13 terpenoids, 14 phenylpropanoids, 4 phenolic acids, 5 phenols, and 11 other phytochemicals. Of these compounds, 17 were confirmed using reference standards. In addition, representative compounds of these different chemical types were used as examples to analyze the fragmentation pathways and characteristic product ions. Moreover, 20 herbs within the QFPD granules were also identified to establish the sources of these chemical components. This is the first rapid profiling of the chemical constituents of QFPD granules using HPLC-Q Exactive Orbitrap-MS and yields valuable information for further quality control and mechanistic studies of QFPD granules.Supplementary InformationThe online version contains supplementary material available at 10.1007/s10337-021-04085-0.

Molecular dynamics simulations reveal the mechanism of the interactions between the inhibitors and SIRT2 at atom level

ABSTRACTThe sirtuins are members of the histone deacetylase family of proteins that participate in a variety of cellular functions and play a close role in cancers. Exploring the proper inhibitors for SIRT2 has attracted great interest in the experiment. However, the detail of the interaction mechanisms between inhibitors and SIRT2 is not well understood. In our study, we synthesised SIRT2 selective inhibitor TPN0_C7 as a model of the proper inhibitor of SIRT2. With the molecular dynamics (MD) simulations, we found that the hydrophobic interactions play the important roles between the inhibitors and SIRT2. According to the conformation character of the inhibitor TPN0_C7, we also explored the natural product, Ge Gen (Puerarol is one of the components.), which is a very effective herb for cancer described in Traditional Chinese Medicine (TCMs) library. Dramatically, we found that the structurally similar inhibitors, Puerarol and TPN0_C7, have the similar binding sites on SIRT2. The hydrophobic and π-π interactions between inhibitors and SIRT2 are important during the progress of the dynamic simulations. In summary, our study uncovers the interaction mechanisms between the inhibitors and SIRT2 at atom level, which may provide clues to explore more proper inhibitors from TCMs.

Structural Insight into the Interactions between Structurally Similar Inhibitors and SIRT6.

Sirtuin 6 (SIRT6) is an NAD+-dependent deacetylase with a significant role in 20% of all cancers, such as colon cancers and rectal adenocarcinoma. However, there is currently no effective drug for cancers related to SIRT6. To explore potential inhibitors of SIRT6, it is essential to reveal details of the interaction mechanisms between inhibitors and SIRT6 at the atomic level. The nature of small molecules from herbs have many advantages as inhibitors. Based on the conformational characteristics of the inhibitor Compound 9 (Asinex ID: BAS13555470), we explored the natural molecule Scutellarin, one compound of Huang Qin, which is an effective herb for curing cancer that has been described in the Traditional Chinese Medicine (TCMS) library. We investigated the interactions between SIRT6 and the inhibitors using molecular dynamics (MD) simulations. We illustrated that the structurally similar inhibitors have a similar binding mode to SIRT6 with residues—Leu9, Phe64, Val115, His133 and Trp188. Hydrophobic and π-stacking interactions play important roles in the interactions between SIRT6 and inhibitors. In summary, our results reveal the interactive mechanism of SIRT6 and the inhibitors and we also provide Scutellarin as a new potential inhibitor of SIRT6. Our study provides a new potential way to explore potential inhibitors from TCMS.

Prim-O-glucosylcimifugin enhances the antitumour effect of PD-1 inhibition by targeting myeloid-derived suppressor cells

BackgroundMyeloid-derived suppressor cells (MDSCs) are immunosuppressive cells that play an important role in immune evasion, PD-1/PD-L1 inhibitor tolerance and tumour progression. Therefore, MDSCs are potential targets for cancer immunotherapy. In this study, we screened an effective polymorphonuclear MDSC (PMN-MDSC) inhibitor from the Traditional Chinese Medicine Library and evaluated its synergistic antitumour effects with PD-1 inhibitor.MethodsIn the present study, we found that PMN-MDSCs accumulate heavily in the spleen and bone marrow of melanoma (B16-F10) tumour-bearing mice. Then, we determined the top 10 key proteins in the upregulated KEGG pathways of PMN-MDSCs in tumour-bearing mice through proteomics and Cytoscape analysis. The key proteins were then used as targets for the screening of PMN-MDSC inhibitors from the traditional Chinese Medicine Library (20000 compounds) through molecular docking and weight calculation of the docking score. Finally, the inhibitory effect of the inhibitor was verified through proteomics and metabolomics analysis in vitro and melanoma (B16-F10) and triple-negative breast cancer (4 T1) mouse tumour models in vivo.ResultsTraditional Chinese medicine saposhnikovia root extract Prim-O-glucosylcimifugin (POG) could bind well to the target proteins and inhibit the proliferation, metabolism and immunosuppressive ability of PMN-MDSCs by inhibiting arginine metabolism and the tricarboxylic acid cycle (TCA cycle). POG could also increase CD8 T-lymphocyte infiltration in the tumours and enhance the antitumour effect of PD-1 inhibitor in B16-F10 and 4 T1 mouse tumour models.ConclusionsPOG was successfully screened from the traditional Chinese Medicine library as a PMN-MDSC inhibitor. POG exhibited a good synergistic antitumour effect with PD-1 inhibitor. This study provided a potential option for enhancing the efficacy of PD-1 inhibitors in clinical applications.

Higenamine, a Dual Agonist for β 1- and β 2-Adrenergic Receptors Identified by Screening a Traditional Chinese Medicine Library.

Chronic heart failure is the terminal stage of various cardiovascular diseases. Despite the availability of several classes of drugs, there is still an unmet need for effective treatment. Based on bench work during the past two decades, we have proposed that enhancement of β 2-adrenergic receptor signaling in combination with the presently preferred β 1-adrenergic receptor blockade would be a promising strategy. Chinese herbal medicines have been shown to be effective in the treatment of heart failure, although the mechanisms largely remain unknown. In the present study, we screened an herbal medicine compound/extract library for β-adrenergic receptor ligands to determine the target of certain effective botanical remedies and seek a leading compound(s) for chronic heart failure treatment. Using a high-throughput screening assay, we identified higenamine, which has a long history in chronic heart failure treatment in traditional Chinese medicine, to be a potent β-adrenergic receptor agonist. Further experiments using specific inhibitors showed that higenamine activated both β 1-adrenergic receptor and β 2-adrenergic receptor. Inhibition of its action by pertussis toxin (a Gi inhibitor) indicated that it is a β 2-adrenergic receptor Gs/Gi dual agonist. Contractility experiments demonstrated a positive inotropic effect of higenamine. In conclusion, we found an herbal compound, higenamine, to be a dual agonist for β 1/β 2-adrenergic receptors with no preference in stimulating the Gs and Gi pathways in β 2-adrenergic receptor signaling. Our results elucidated not only the target of higenamine to explain its pharmacological effect in treating chronic heart failure, but also the mechanisms of its cardiac toxicity.

Structure-Based in Silico Screening Identifies a Potent Ebolavirus Inhibitor from a Traditional Chinese Medicine Library.

Potent Ebolavirus (EBOV) inhibitors will help to curtail outbreaks such as that which occurred in 2014–16 in West Africa. EBOV has on its surface a single glycoprotein (GP) critical for viral entry and membrane fusion. Recent high-resolution complexes of EBOV GP with a variety of approved drugs revealed that binding to a common cavity prevented fusion of the virus and endosomal membranes, inhibiting virus infection. We performed docking experiments, screening a database of natural compounds to identify those likely to bind at this site. Using both inhibition assays of HIV-1-derived pseudovirus cell entry and structural analyses of the complexes of the compounds with GP, we show here that two of these compounds attach in the common binding cavity, out of eight tested. In both cases, two molecules bind in the cavity. The two compounds are chemically similar, but the tighter binder has an additional chlorine atom that forms good halogen bonds to the protein and achieves an IC50 of 50 nM, making it the most potent GP-binding EBOV inhibitor yet identified, validating our screening approach for the discovery of novel antiviral compounds.

Aldehyde dehydrogenase 3A1 activation prevents radiation-induced xerostomia by protecting salivary stem cells from toxic aldehydes

Xerostomia (dry mouth) is the most common side effect of radiation therapy in patients with head and neck cancer and causes difficulty speaking and swallowing. Since aldehyde dehydrogenase 3A1 (ALDH3A1) is highly expressed in mouse salivary stem/progenitor cells (SSPCs), we sought to determine the role of ALDH3A1 in SSPCs using genetic loss-of-function and pharmacologic gain-of-function studies. Using DarkZone dye to measure intracellular aldehydes, we observed higher aldehyde accumulation in irradiated Aldh3a1-/- adult murine salisphere cells and in situ in whole murine embryonic salivary glands enriched in SSPCs compared with wild-type glands. To identify a safe ALDH3A1 activator for potential clinical testing, we screened a traditional Chinese medicine library and isolated d-limonene, commonly used as a food-flavoring agent, as a single constituent activator. ALDH3A1 activation by d-limonene significantly reduced aldehyde accumulation in SSPCs and whole embryonic glands, increased sphere-forming ability, decreased apoptosis, and improved submandibular gland structure and function in vivo after radiation. A phase 0 study in patients with salivary gland tumors showed effective delivery of d-limonene into human salivary glands following daily oral dosing. Given its safety and bioavailability, d-limonene may be a good clinical candidate for mitigating xerostomia in patients with head and neck cancer receiving radiation therapy.

Identification of cardioprotective agents from traditional Chinese medicine against oxidative damage.

Reactive oxygen species are damaging to cardiomyocytes. H9c2 cardiomyocytes are commonly used to study the cellular mechanisms and signal transduction in cardiomyocytes, and to evaluate the cardioprotective effects of drugs following oxidative damage. The present study developed a robust, automated high throughput screening (HTS) assay to identify cardioprotective agents from a traditional Chinese medicine (TCM) library using a H2O2-induced oxidative damage model in H9c2 cells. Using this HTS format, several hits were identified as cardioprotective by detecting changes to cell viability using the cell counting kit (CCK)-8 assay. Two TCM extracts, KY-0520 and KY-0538, were further investigated. The results of the present study demonstrated that treatment of oxidatively damaged cells with KY-0520 or KY-0538 markedly increased the cell viability and superoxide dismutase activity, decreased lactate dehydrogenase activity and malondialdehyde levels, and inhibited early growth response-1 (Egr-1) protein expression. The present study also demonstrated that KY-0520 or KY-0538 treatment protected H9c2 cells from H2O2-induced apoptosis by altering the Bcl-2/Bax protein expression ratio, and decreasing the levels of cleaved caspase-3. In addition, KY-0520 and KY-0538 reduced the phosphorylation of ERK1/2 and p38-MAPK proteins, and inhibited the translocation of Egr-1 from the cytoplasm to nucleus in H2O2-treated H9c2 cells. These findings suggested that oxidatively damaged H9c2 cells can be used for the identification of cardioprotective agents that reduce oxidative stress by measuring cell viabilities using CCK-8 in an HTS format. The underlying mechanism of the cardioprotective activities of KY-0520 and KY-0538 may be attributed to their antioxidative activity, regulation of Egr-1 and apoptosis-associated proteins, and the inhibition of ERK1/2, p38-MAPK and Egr-1 signaling pathways.

Abstract 4556: Identification of novel agent for HCC treatment by GNMT-promoter orientated cell-based drug screening platform

Abstract Glycine N-methyltransferase (GNMT) a tumor suppressor gene for hepatocellular carcinoma (HCC) plays very important role in liver detoxification and tumorigenisis. GNMT expression is highly down-regulated in HCC. Recent studies have shown that overexpression of GNMT reduce tumor growth and sensitize HuH-7 cells to rapamycin treatment in vitro and in vivo. Therefore, enhancing GNMT expression is an attractive approach for HCC therapy. In present study we developed a GNMT-promoter driven luciferase reporter assay as a drug screen platform for the identification of novel drugs for prevention and treatment of HCC. We identified crude extract 317 as highest activator of GNMT promoter activity from a Chinese medicine library containing 472 herbal extracts. Partial purified fraction “F2-6” was isolated by bioassay-guided fractionation from crude extract 317 and validated for enhancement of endogenous GNMT expression at both mRNA and protein levels. Fraction F2-6 inhibited growth of liver cancer cell lines in vitro. Moreover, orally administrated F2-6 can delay and reduce growth of HCC xenograft tumor in vivo. Next, we identified the active component in fraction F2-6 named as compound P. Furthermore, we demonstrated that combination of sorafenib with compound P led to additive toxicity on HCC cells in vitro and in vivo. The cDNA microarray and real time PCR analysis showed that compound P altered cell cycle-associated genes such as CDK6, CCND1, CDKN1C, MYC and PLK1. Finally, Compound P treatment induced apoptosis in huh7 cells were confirmed by increased sub-G1 population and caspase 3/7 activity. Collectively, our results suggest that compound P has potential to be a novel agent for prevention and treatment of HCC. Citation Format: Rajni Kant, Lucas Tung, Chung-Kuang Lu, Yi-Ming Chen, Chia-Hung Yen. Identification of novel agent for HCC treatment by GNMT-promoter orientated cell-based drug screening platform. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4556. doi:10.1158/1538-7445.AM2015-4556

Methods on application of Chinese medicine to protect ancient books

Objective Pest control is one of the measures in protecting ancient books.To conclude pest control methods of Heilongjiang University of Chinese Medicine Library,a suitable method for promotion can be found.Methods By integrating the practical experience of protecting ancient books in Heilongjiang University,this article discussed concrete applications of Lysimachia foenum-gracum Hance to protect ancient books from infestation.Results The pest control mechanism of Lysimachia foenum-gracum was analyzed.The best pest control effects can be achieved when we identify the origin,choose the right picking season,identify authenticity of the medicine,and correctly use it.Conclusion Using Chinese traditional medicine to keep away insect is an environmental friendly way,which is worthy of generalizing. Key words: Lysimachia foenum-gracum; Chinese Medicine; Protection of ancient books on traditional Chinese medicine

The challenges and development strategies of traditional Chinese medicine professional library in the era of big data

With the rapid development of computer technology and communication technology,especially with the rapid rise and popularity of the technologies on Internet,Internet of Things and Cloud Computing,the variety of data produced by the human society grow exponentially,the human society has entered the era of Big Data.This paper summarizes the concepts,features and strategic significances of Big Data,and studies the Big Data characteristics of the library,discusses the challenges of library and the development strategies of professional library in the era of Big Data with the example of traditional Chinese medicine professional library. Key words: Big data ; Library ; Traditional Chinese medicine library ; Knowledge services ; Cloud computing ; Cloud services of digital library ;