Inhibition of multiple SARS-CoV-2 variants entry by Lycium barbarum L. polysaccharides through disruption of spike protein-ACE2 interaction.

Viral respiratory infections are major human health concerns. The most striking epidemic disease, COVID-19 is still on going with the emergence of fast mutations and drug resistance of pathogens. A few polysaccharide macromolecules from traditional Chinese medicine (TCM) have been found to have direct anti-SARS-CoV-2 activity but the mechanism remains unclear. In this study, we evaluated the entry inhibition effect of Lycium barbarum polysaccharides (LBP) in vitro and in vivo. We found LBP effectively suppressed multiple SARS-CoV-2 variants entry and protected K18-hACE2 mice from invasion with Omicron pseudovirus (PsV). Moreover, we found LBP interfered with early entry events during infection in time-of-addition (TOA) assay and SEM observation. Further surface plasmon resonance (SPR) study revealed the dual binding of LBP with Spike protein and ACE2, which resulted in the disruption of Spike-ACE2 interaction and subsequently triggered membrane fusion. Therefore, LBP may act as broad-spectrum inhibitors of virus entry and nasal mucosal protective agent against newly emerging respiratory viruses, especially SARS-CoV-2.

Degradative solvent extraction of cyanobacteria: From reaction kinetics to potential organic matter evolution mechanism.

This study proposed a new continuous lumped reaction kinetics model to accurately reveal the control mechanism of cyanobacteria at each stage of degradative solvent extraction and discussed the potential evolution mechanism of organic matter. Results showed that degradation solvent extraction successfully separated nitrogen and phosphorus from cyanobacteria. The solute has high carbon and volatile contents, is almost ash-free, and forms a phosphorus-rich residue. The lowest fitting degree of the continuous lumped reaction model kinetics was 94.5%, suggesting that this model worked well. The depolymerization of the residue dominated between 200 and 350 °C, whereas solute decomposition dominated at 400 °C. Nitrogen-containing compounds, which originate from protein decarboxylation or deamination to generate amides, are the main components of the solute, and amino acids react with reducing sugars to generate nitrogen heterocyclic compounds, which are useful for preparing nitrogen-containing chemicals.

Synergistic Antibacterial Surface with Liquid Repellency and Enhanced Photothermal Bactericidal Activity.

The presence of microorganisms on biomedical devices and food packaging surfaces poses an important threat to human health. Superhydrophobic surfaces, a powerful tool to combat pathogenic bacterial adhesion, are threatened by their poor robustness. As a supplement, photothermal bactericidal surfaces may be expected to kill adhered bacteria. Using copper mesh as a mask, we prepared a superhydrophobic surface with a homogeneous conical array. The surface shows synergistic antibacterial properties, including a superhydrophobic character against bacterial adhesion and photothermal bactericidal activity. As a result of the excellent liquid repellency, the surface could highly repel the adherence of bacteria after immersing in a bacterial suspension for 10 s (95%) and 1 h (57%). Photothermal graphene can easily eliminate most adhered bacteria during the subsequent treatment of near-infrared (NIR) radiation. After a self-cleaning wash, the deactivated bacteria were easily rinsed off the surface. Furthermore, this antibacterial surface exhibited an approximately 99.9% resisted bacterial adhesion rate regardless of planar and various uneven surfaces. The results offer promising advancement of an antibacterial surface combining both adhesion resistance and photothermal bactericidal activity in fighting microbial infections.

Potent insecticidal activity of Eleocharis dulcis (Burm. f.) Trin peel extract and its main components against aphids.

BACKGROUND: Aphids are significant pests of cash crops and food farm crops. Botanical insecticides are safe for aphid control, especially for organic farming. In this study, Eleocharis dulcis (Burm. f.) Trin. peel extract (EDPE), a new botanical insecticide, was investigated for its active compositions against several agricultural aphids. RESULTS: The results showed that the EDPE had high insecticidal activity against Sitobion avenae Fabricius, Aphis gossypii Glover, Megoura crassicauda Mordvilko, and Acyrthosiphon pisum Harris, with half-lethal concentration (LC50 ) values of 95.92, 81.04, 140.31, and 255.73 mg/L after 48 h of treatment. In the pot culture assay, the aphicidal effects of 25% EDPE soluble liquid (SL) at a concentration of 0.016% were 68.98 ± 5.61%, 79.33 ± 8.27%, and 88.82 ± 3.91% after the first, third, and seventh days of treatment, respectively. Nine compounds were identified by bioactivity-directed fractionation: 4',5'-dimethoxy-6,6-dimethylpyranoisoflavone (1), 3-methoxy-4-hydroxylonchocarpin (2), 4-hydroxylonchocarpin (3), 4-methoxylonchocarpin (4), barbigerone (5), lonchocarpusone (6), 6a,12a-dehydrodeguelin (7), 13-homo-13-oxa-6a, 12a-dehydrodeguelin (8) and deguelin (9). Among them, 4-hydroxylonchocarpin (3) showed the highest aphidicidal activity against M. crassicauda, S. avenae, and A. pisum, with LC50 values of 97.24, 140.63, and 112.31 mg/L, respectively. CONCLUSION: These data contribute to a better understanding of the aphicidal activity of EDPE and its main component, 4-hydroxylonchocarpin. This will help to develop new botanical insecticides to contro aphids. © 2022 Society of Chemical Industry.

Salt reducing and saltiness perception enhancing strategy for shiitake (Lentinus edodes) bud using novel combined treatment of yeast extract and radio frequency.

This study investigated the potential of yeast extract and radio frequency (RF) treatment as a strategy of reducing salt and enhancing saltiness perception for Lentinus edodes bud. The results of E-nose demonstrated yeast extract and RF treatment improved the saltiness of Lentinus edodes bud. Meanwhile, yeast extract and RF treatment significantly decreased the addition of salt (P < 0.05), and led to the formation of special flavor substances, whereas amino acid nitrogen content decreased. On the other hand, sensory attribute, hardness, total flavonoid and phenolic content, antioxidant capacity of L. edodes buds significantly (P < 0.05) increased after the yeast extract combined with RF treatment. In addition, the modification of water distribution, the formation of dense structure, uniform microstructure and Na+ distribution were observed in treated sample, causing the enhancement of saltiness perception. Accordingly, the alteration of properties contributed to higher sensory properties of texture, taste, flavor, and overall acceptability.

Antibacterial mechanisms of star anise essential oil microcapsules encapsulated by rice protein-depolymerized pectin electrostatic complexation and its application in crab meatballs.

A mixed solid matrix of the depolymerized pectin (DP) and rice protein (RP) were investigated to improve the antibacterial activity of star anise essential oil (SAEO) through microencapsulation. The encapsulation was undertaken via electrostatic complexation method followed by spray drying. Under the optimized conditions, SAEO microcapsules with 92.2 % encapsulation efficiency was acquired at 3:2 of DP-to-RP, 1:3 of DP-to-SAEO and pH 4.0. DP-RP/SAEO microcapsules exhibited a spherical-shaped particle with smaller particle size, and sustained release. FTIR and morphology analysis confirmed that SAEO was successfully encapsulated in the solid matrix of microcapsules. DP-RP/SAEO microcapsules caused the destruction of cytomembranes and reduction of membrane proteins, which led to the alteration of cell membrane fluidity and integrity. Meanwhile, DP-RP/SAEO microcapsules repressed the key enzyme in tricarboxylic acid (TCA) and Hexose monophosphate pathway (HMP) cycle pathway of E. coli, S. aureus. The application experiments indicated DP-RP/SAEO microcapsules can effectively control the growth of E. coli and S. aureus in crab meatballs.

Acupuncture for Pain and Function in Patients with Nonspecific Low Back Pain: Study Protocol for an Up-to-Date Systematic Review and Meta-Analysis.

Aim: To assess the pain and function of acupuncture in patients with nonspecific low back pain (NSLBP). Design: This is a study protocol for an upcoming, updated systematic review and meta-analysis. We have registered on the PROSPERO platform, and the registration number is CRD42020149647. Methods: We will identify studies in seven English databases, including Web of Science, Cochrane Library, CINAHL, Physiotherapy Evidence Database, Medline, Embase, and Scopus, and in four Chinese databases, including China National Knowledge Infrastructure (CNKI), China Biology Medicine disc (CBMdisc), Wanfang Database, and VIP database. We will also manually search key journals about acupuncture, the references of identified relevant trials, and the Clinical Trials Registry Platform. Different search strategies of Medical Subject Headings (MeSH) and non-MeSH terms will be applied alone or integrated. We will evaluate methodological quality by adopting Cochrane's risk of bias criteria and perform a meta-analysis using RevMan V.5.4 software. Results: This study's results will be reported according to the PRISMA checklist and presented at an authoritative scientific conference or an authoritative scientific journal.

Mechanisms of Compound Kushen Injection for the treatment of bladder cancer based on bioinformatics and network pharmacology with experimental validation.

Bladder cancer is the most common malignancy of the urinary system. Compound Kushen Injection (CKI) is a Chinese medicinal preparation that has been widely used in the treatment of various types of cancers in the past two decades. However, the pharmacological effect of CKI on bladder cancer is not still completely understood. In the current study, network pharmacology combined with bioinformatics was used to elucidate the therapeutic mechanism and potential targets of CKI in bladder cancer. The mechanism by which CKI was effective against bladder cancer was further verified in vitro using human bladder cancer cell line T24. Network pharmacology analysis identified 35 active compounds and 268 target genes of CKI. Bioinformatics data indicated 5500 differentially expressed genes associated with bladder cancer. Common genes of CKI and bladder cancer suggested that CKI exerted anti-bladder cancer effects by regulating genes such as MMP-9, JUN, EGFR, and ERK1. Functional enrichment analysis indicated that CKI exerted therapeutic effects on bladder cancer by regulating certain biological processes, including cell proliferation, cell migration, and cell apoptosis. In addition, Kyoto Encyclopedia of Genes and Genomes enrichment analysis implicated pathways related to cancer, bladder cancer, and the PI3K-Akt signaling pathway. Consistently, cell experiments indicated that CKI inhibited the proliferation and migration of T24 cells, and induced their apoptosis. Moreover, RT-qPCR and Western blot results demonstrated that CKI was likely to treat bladder cancer by down-regulating the gene and protein expression of MMP-9, JUN, EGFR, and ERK1. CKI inhibited the proliferation and migration, and induced the apoptosis of T24 bladder cancer cells through multiple biological pathways and targets. CKI also exhibited significant effects on the regulation of key genes and proteins associated with bladder cancer. Overall, our findings provide solid evidence and deepen current understanding of the therapeutic effects of CKI for bladder cancer, and further support its clinical use.

Salvia miltiorrhiza bunge exerts anti-oxidative effects through inhibiting KLF10 expression in vascular smooth muscle cells exposed to high glucose.

ETHNOPHARMACOLOGICAL RELEVANCE: Traditional Chinese medicinal herb Salvia miltiorrhiza Bunge(Danshen) and its components have been widely used to treat cardiovascular diseases for hundreds of years in China, including hypertension, diabetes, atherosclerosis, and chronic heart failure. Salvia miltiorrhiza injection (SMI), an aqueous extracts of Salvia miltiorrhiza Bunge, is one of most widely used traditional Chinese medicine injections. SMI is widely used in the treatment of diabetic vascular complications, However, the mechanisms remain to be defined. AIM OF THE STUDY: To investigate protective mechanism of Salvia miltiorrhiza Bunge against ROS generation in VSMCs of diabetic mice and patients. MATERIALS AND METHODS: Salvia miltiorrhiza injection (hereinafter referred to as SMI, 1.5 g mL-1), which was approved by the State Food and Drug Administration (approval number: Z32020161), was obtained from Shenlong Pharmaceutical Co., Ltd. (batch number: 11040314). SMI or vehicle were intraperitoneally administrated to the HFD-fed db/db mice, artery was harvested after 24weeks later. qRT-PCR and Western blot analysis were used to detect the expression of KLF6, KLF5, KLF4, KLF10, KLF12, and HO-1. DCFH-DA staining detected intracellular ROS production. Loss- and gain-of-function experiments of KLF10 were used to investigate the effect of KLF10 on the expression of HO-1. Dual-luciferase reporter assay evaluated the effect of KLF10 on the activity of the HO-1 promoter. RESULTS: KLF10 expression and ROS generation are significantly increased in the arteries of HFD-fed db/db mice, VSMCs of diabetic patients, as well as in high glucose-treated VSMCs. KLF10 overexpression suppresses, while its knockdown facilitates the expression of heme oxygenase (HO-1) mRNA and protein. Further, Salvia miltiorrhiza injection (SMI) abrogates KLF10 upregulation and reduces ROS generation induced by high glucose in VSMCs. Mechanistically, KLF10 negatively regulates the HO-1 gene transcription via directly binding to its promoter. Accordingly, SMI treatment of VSMCs reduces ROS generation through inhibiting KLF10 expression and thus relieving KLF10 repression of the expression of HO-1 gene, subsequently contributing to upregulation of HO-1. CONCLUSION: SMI exerts anti-oxidative effects on VSMCs exposed to high glucose through inhibiting KLF10 expression and thus upregulating HO-1.

Influence of high hydrostatic pressure pretreatment on properties of vacuum-freeze dried strawberry slices.

Effects of high hydrostatic pressure (HHP) (50, 100, 150, 200 and 250 MPa) pretreatment on water mobility and distribution, drying duration, microstructure, color, cell wall fraction and tissue structure of strawberry slices were investigated. HHP significantly increased water mobility of the strawberry slices, resulting in the reduction of drying duration by 9-24%. As the pretreatment pressure was increased, redness value and anthocyanin content continuously increased, soluble pectin (SBP) content increased and then decreased, while the contents of protopectin (PTP) and cellulose decreased. After the HHP pretreatment, chromoplasts and moisture was distributed more uniformly in the strawberry slices. Microscopy images showed the formation of microscopic holes or channels in the matrix and the breakdown of tissue structure by HHP. Results suggested HHP pretreatment disrupted the integrity of the fresh strawberry which enhanced the drying efficiency and migration of the chromoplasts during the vacuum-freeze drying process.

A network pharmacology approach to investigate the mechanism of Shuxuening injection in the treatment of ischemic stroke.

ETHNOPHARMACOLOGICAL RELEVANCE: Shuxuening injection (SXNI), a popular herbal medicine, is an extract of Ginkgo biloba leaves (GBE), and is used to treat ischemic stroke (IS) in China. However, its specific active ingredients and molecular mechanisms in IS remain unclear. AIM OF THE STUDY: The purpose of the research is to identify the main active ingredients in GBE and explore its molecular mechanisms in the treatment of IS. MATERIALS AND METHODS: The main active components of GBE were discerned through the Traditional Chinese Medicine Systems Pharmacology Database and Analysis (TCMSP), Traditional Chinese Medicine Integrated Database (TCMID), Bioinformatics Analysis Tool for Molecular Mechanism of Traditional Chinese Medicine (BATMAN-TCM) database, and absorption, distribution, metabolism and excretion (ADME) analysis. The targets related to IS were obtained using Genecards, Online Mendelian Inheritance in Man (OMIM), Therapeutic Target Database (TTD), and Disgenet. We discovered an intersection of genes. Subsequently, protein-protein interaction (PPI) networks were constructed with Cytoscape 3.7.1 and the String database. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses were performed to analyze the intersection of targets via the Database for Annotation, Visualization, and Integrated Discovery (DAVID) 6.8. Built on the above analysis, we made a Compound-Target-Pathway (C-T-P) network. Autodock Vina was used for molecular docking analysis. Maestro 11.9 was used to calculate the root-mean-square deviation (RMSD). Animal experiments were performed to verify the core targets. Triphenyl tetrazolium chloride (TTC) staining was used to calculate the infarct volume in rats. Hematoxylin-eosin (HE) staining was employed to observe the morphology of hippocampal neuron cells. RT-qPCR was applied to detect relative mRNA levels, and protein expression was determined using Western blotting. RESULTS: Molecular docking showed that PTGS2, NOS3 and CASP3 docked with small molecule compounds. According to RT-qPCR and Western blotting, mRNA and protein expression of PTGS2 and CASP3 were up-regulated (P < 0.05), and mRNA and protein levels of NOS3 were down-regulated (P < 0.05). CONCLUSIONS: SXNI can treat IS through multiple targets and routes, and reduce the apoptosis of neuron cells in brain tissue by inhibiting inflammation and regulating the level of oxidative stress, thereby protecting rats brain tissue.

Tea consumption and risk of diabetes in the Chinese population: a multi-centre, cross-sectional study.

The aim of the present study was to explore the influence of tea consumption on diabetes mellitus in the Chinese population. This multi-centre, cross-sectional study was conducted in eight sites from south, east, north, west and middle regions in China by enrolling 12 017 subjects aged 20-70 years. Socio-demographic and general information was collected by a standardised questionnaire. A standard procedure was used to measure anthropometric characteristics and to obtain blood samples. The diagnosis of diabetes was determined using a standard 75-g oral glucose tolerance test. In the final analysis, 10 825 participants were included and multiple logistic models and interaction effect analysis were applied for assessing the association between tea drinking with diabetes. Compared with non-tea drinkers, the multivariable-adjusted OR for newly diagnosed diabetes were 0·80 (95 % CI 0·67, 0·97), 0·88 (95 % CI 0·71, 1·09) and 0·86 (95 % CI 0·67, 1·11) for daily tea drinkers, occasional tea drinkers and seldom tea drinkers, respectively. Furthermore, drinking tea daily was related to decreased risk of diabetes in females by 32 %, elderly (>45 years) by 24 % and obese (BMI > 30 kg/m2) by 34 %. Moreover, drinking dark tea was associated with reduced risk of diabetes by 45 % (OR 0·55; 95 % CI 0·42, 0·72; P < 0·01). The results imply that drinking tea daily was negatively related to risk of diabetes in female, elderly and obese people. In addition, drinking dark tea was associated with decreased risk of type 2 diabetes mellitus.

Identifying possible non-thermal effects of radio frequency energy on inactivating food microorganisms.

Radio frequency (RF) heating has been successfully used for inactivating microorganisms in agricultural and food products. Athermal (non-thermal) effects of RF energy on microorganisms have been frequently proposed in the literature, resulting in difficulties for developing effective thermal treatment protocols. The purpose of this study was to identify if the athermal inactivation of microorganisms existed during RF treatments. Escherichia coli and Staphylococcus aureus in apple juice and mashed potato were exposed to both RF and conventional thermal energies to compare their inactivation populations. A thermal death time (TDT) heating block system was used as conventional thermal energy source to simulate the same heating treatment conditions, involving heating temperature, heating rate and uniformity, of a RF treatment at a frequency of 27.12 MHz. Results showed that a similar and uniform temperature distribution in tested samples was achieved in both heating systems, so that the central sample temperature could be used as representative one for evaluating thermal inactivation of microorganisms. The survival patterns of two target microorganisms in two food samples were similar both for RF and heating block treatments since their absolute difference of survival populations was <1 log CFU/ml. The statistical analysis indicated no significant difference (P > 0.05) in inactivating bacteria between the RF and the heating block treatments at each set of temperatures. The solid temperature and microbial inactivation data demonstrated that only thermal effect of RF energy at 27.12 MHz was observed on inactivating microorganisms in foods.

Continuous liquid fermentation of pretreated waste activated sludge for high rate volatile fatty acids production and online nutrients recovery.

Raw sludge was pretreated, and the separated sludge liquid was used as substrate in a continuous operated up-flow anaerobic sludge blanket reactor to produce volatile fatty acids (VFAs). The highest VFA productivity of continuous fermentation with sludge liquid at an organic loading rate (OLR) of 10.0 kg COD/m3/d was about 5.0-fold and 4.0-fold higher than batch and semi-continuous fermentation with pretreated sludge slurry, respectively. Moreover, the liquid fermentation with an OLR of 10.0 kg COD/m3/d consumed the least energy, which was about 10.57% and 12.12% of batch and semi-continuous sludge fermentation, respectively. When combined with online nitrogen and phosphorus recovery, VFA production further increased by 20.67% and struvite recovery efficiency reached 1.98  ±â€¯â€¯0.28 g/g PO43-. The process showed high VFA production, low energy consumption and good nutrients recovery by continuous liquid anaerobic fermentation, significantly increasing the economic potential of VFA production from waste activated sludge.

Verification of radio frequency pasteurization treatment for controlling Aspergillus parasiticus on corn grains.

Radio frequency (RF) heating has been proposed and tested to achieve a required anti-fungal efficacy on various food samples due to its advantage of deeper penetration depth and better heating uniformity. The purpose of this study was to validate applications of RF treatments for controlling Aspergillus parasiticus in corn while maintaining product quality. A pilot-scale, 27.12MHz, 6kW RF heating system together with hot air heating was used to rapidly pasteurize 3.0kg corn samples. Results showed that the pasteurizing effect of RF heating on Aspergillus parasiticus increased with increasing heating temperature and holding time, and RF heating at 70°C holding in hot air for at least 12min resulted in 5-6 log reduction of Aspergillus parasiticus in corn samples with the moisture content of 15.0% w.b. Furthermore, thermal resistance of Aspergillus parasiticus decreased with increasing moisture content (MC) of corn samples. Quality (MC, water activity - aw, protein, starch, ash, fat, fatty acid, color, electrical conductivity and germination rate) of RF treated corn met the required quality standard used in cereal industry. Therefore, RF treatments can provide an effective and rapid heating method to control Aspergillus parasiticus and maintain acceptable corn quality.

Effects of troxerutin on cognitive deficits and glutamate cysteine ligase subunits in the hippocampus of streptozotocin-induced type 1 diabetes mellitus rats.

Increasing evidence demonstrates an association between diabetes and hippocampal neuron damage. This study aimed to determine the effects of troxerutin on cognitive deficits and glutamate cysteine ligase subunits (GCLM and GCLC) in the hippocampus of streptozotocin-induced type 1 diabetes mellitus (T1DM) rats. At 12weeks after streptozotocin injection, T1DM rats were randomly divided into 4 groups (n=15 each group) to receive no treatment (T1DM), saline (T1DM+saline), alpha-lipoic acid (T1DM+alpha-lipoic acid), and troxerutin (T1DM+troxerutin), respectively, for 6weeks. Meanwhile, 10 control animals (NC group) were assessed in parallel. Learning performance was evaluated by the Morris water maze. After treatment, hippocampi were collected for pathological examination by hematoxylin and eosin (H&E) staining. Next, hippocampal superoxide dismutase (SOD) activity, and malondialdehyde (MDA) and glutathione (GSH) levels were assessed. Finally, glutamate cysteine ligase catalytic (GCLC) and glutamate cysteine ligase modifier (GCLM) subunit mRNA and protein levels were quantified by reverse transcription polymerase chain reaction (RT-PCR) and Western blot, respectively. Compared with T1DM and T1DM+saline groups, escape latency was overtly reduced in T1DM+alpha-lipoic acid and T1DM+troxerutin groups. Significantly increased GCLM and GCLC mRNA levels, GCLC protein amounts, SOD activity, and GSH levels, and reduced MDA amounts were observed in T1DM+alpha-lipoic acid and T1DM+troxerutin groups. In T1DM and T1DM+saline groups, H&E staining showed less pyramidal cells in the hippocampus, with disorganized layers, karyopyknosis, decreased endochylema, and cavitation, effects relieved in T1DM+alpha-lipoic acid and T1DM+troxerutin groups. Troxerutin alleviates oxidative stress and promotes learning in streptozotocin-induced T1DM rats, a process involving GCLC expression.

Screening and identification of hepatotoxic component in Evodia rutaecarpa based on spectrum-effect relationship and UPLC-Q-TOFMS.

Evodia rutaecarpa (E. rutaecarpa) has been used to treat aches, vomiting and dysentery in traditional Chinese medicine. However, as a mildly toxic herb its toxic components have not been elucidated. An attempt was made to illuminate the hepatotoxic constituents of E. rutaecarpa. The 50% ethanol extracts of E. rutaecarpa from 19 different sources were used to establish UPLC fingerprints and administered to mice at a dose of 35 g/kg (crude medicine weight/mouse weight) once daily for 14 days. Serum levels of alanine transaminase, aspartate aminotransferase and liver coefficient were used as indices of liver injury. Additionally, the characteristic peaks of 19 fingerprints were identified. Spectrum-effect relationships between fingerprints and hepatotoxic indicators were analyzed using bivariate correlation analysis (BCA). The UPLC fingerprints were established and a total of 28 main compounds were identified. Because of the inherent variations in chemical compositions, the liver injury levels were different among the E. rutaecarpa samples from 19 sites of production. BCA results indicated that compounds dihydrorutaecarpine, 6-acetoxy-5-epilimonin, goshuyuamide I, 1-methyl-2-[(Z)-5-undecenyl]-4(1H)-quinolone, 1-methyl-2-[(4Z,7Z)-4,7-tridecadienyl]-4(1H)-quinolone, evocarpine and 1-methyl-2-[(6Z,9Z)-6,9-pentadecadienyl]-4(1H)-quinolone were tentatively determined as the primary hepatotoxic components. The present study provides a valuable method for the discovery of hepatotoxic constituents by combination of fingerprints and hepatotoxicity index.

Aurantiamide acetate suppresses the growth of malignant gliomas in vitro and in vivo by inhibiting autophagic flux.

We aim to investigate the effect of aurantiamide acetate isolated from the aerial parts of Clematis terniflora DC against gliomas. Human malignant glioma U87 and U251 cells were incubated with different concentrations (0-100 µM) of aurantiamide acetate. Aurantiamide acetate greatly decreased the cell viability in a dose- and time-dependent manner. It induced moderate mitochondrial fragmentation and the loss of mitochondrial membrane potential. No significant difference was found in the alternation of other intracellular organelles, although F-actin structure was slightly disturbed. Apparent ultrastructure alternation with increased autophagosome and autolysosome accumulation was observed in aurantiamide acetate-treated cells. The expression of LC3-II was greatly up-regulated in cells exposed to aurantiamide acetate (P < 0.05 compared with control). The cytoplasmic accumulation of autophagosomes and autolysosomes induced by aurantiamide acetate treatment was confirmed by fluorescent reporter protein labelling. Administration of chloroquine (CQ), which inhibits the fusion step of autophagosomes, further increased the accumulation of autophagosomes in the cytoplasm of U87 cells. Autophagy inhibition by 3-methyladenine, Bafilomycin A1 or CQ had no influence on aurantiamide acetate-induced cytotoxicity, whereas autophagy stimulator rapamycin significantly suppressed aurantiamide acetate-induced cell death. The anti-tumour effects of aurantiamide acetate were further evaluated in tumour-bearing nude mice. Intratumoural injection of aurantiamide acetate obviously suppressed tumour growth, and increased number of autophagic vacuoles was observed in tumour tissues of animals receiving aurantiamide acetate. Our findings suggest that aurantiamide acetate may suppress the growth of malignant gliomas by blocking autophagic flux.