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Two new chemical constituents, japopenoid D (1), and japopenoid E (2), were isolated and identified from the flower buds of Lonicera japonica Thunb. The structures of these compounds were elucidated based on spectroscopic analysis (HR-ESI-MS, NMR), and the absolute configurations of 1 and 2 were determined by comparison of their electronic circular dichroism (ECD) spectra with literature and theoretical calculation. The anti-inflammatory activities of the isolates were evaluated by measuring their inhibitory effects on PGE2 and IL-6 production in LPS stimulated RAW 264.7 macrophages. As a result, compound 1 could reduce PGE2 and IL-6 levels in LPS-activated RAW 264.7 macrophages with IC50 values of 6.78 and 42.07 μM, respectively.

Eight previously undescribed diarylheptanoids (1–8), together with fifteen known analogues (9–23), were isolated from the rhizomes of Zingiber officinale. Their structures were unambiguously determined by comprehensive spectroscopic analyses and electronic circular dichroism (ECD) calculations. It is worth mentioning that 1–3 are the first reported structures of diaryl ether heptanoids in Z. officinale, whereas 15–17 were isolated from Zingiber for the first time. Furthermore, a gene enrichment analysis of the interacting targets indicated that diarylheptanoids were mainly associated with the anti-tumor activity by affecting DNA damage signaling pathway. The results showed that 6, 16–19 had remarkable inhibitory effects against five tumor cell lines (A549, HepG2, HeLa, MDA-MB-231, and HCT116) with IC50 values ranging from 6.69–33.46 μM, and showed down-regulating the expression of ATR (ataxia telangiectasia mutated and RAD3-related) and CHK1 (checkpoint kinase 1) levels in HCT116 and A549 cell lines. Our studies not only enrich the structural diversity of diarylheptanoids in nature, but also discover several natural compounds for anti-tumor agents.

Phytochemical investigations on the dry leaves of Illicium dunnianum have led to the isolation of 24 lignans. Illiciumlignans G–K (1–5) were five undescribed benzofuran lignans, illiciumlignan L (6) was one undescribed ditetrahydrofuran lignan, illiciumlignans M–O (7–9) were three new sesquilignans, and compounds 10, 12, 13, 15, and 18–21 were firstly isolated from the genus Illicium. Their structures were elucidated by detailed spectroscopic analyses (UV, IR, HR-ESI-MS, and NMR) and CD experiments. All isolates were evaluated by measuring their inhibitory effects on PGE2, and NO production in LPS-stimulated RAW 264.7 macrophages.

Cadmium (Cd) as a non‐essential toxic metal has become one of the seriously environmental problems. Overload of Cd into plant shoots, particularly the addible parts (i.e., grains), jeopardizes crop production and food safety. Isolating and identifying genotypic variations in Cd accumulation of rapeseed (Brassica napus) cultivars is an efficient approach for phytoremediation and developing lower Cd‐accumulating plants. In this study, a trial was conducted under natural condition in Nanjing, China, from 2014 to 2017, and identified 64 rapeseed cultivars collected from the areas of Gui Zhou province. Rapeseed grew under moderate Cd exposure (5 mg kg⁻¹) for 5 months, and shoots were harvested for Cd quantification. A great variation of total Cd concentrations in shoots, ranking from 0.16 to 17.03 mg Cd kg⁻¹, was found. Following the initial examination of all cultivars, two sets of plants with high (#138 and #177) and low (#208 and #244) Cd concentrations were further investigated. Throughout the growth period, cultivars #138 and #177 accumulated more Cd during vegetative (30, 60, and 120 d) and late developmental (180 d) stages than cultivars #208 and #244. The higher Cd concentration in shoots of #138 and #177 was associated with the higher Cd concentration in xylem sap, suggesting the greater capability of Cd translocation from roots to shoots. Compared to #208 and #244, Cd exposure moderately reduced zinc and iron concentrations in some tissues of #138 and #177, whereas the manganese and magnesium concentrations showed no change. Although #138 and #177 cultivars accumulated more Cd in their shoots, no Cd toxicity was detected. Moreover, both #138 and #177 cultivars had a similar biomass to #208 or #244. These results suggest that #138 and #177 rapeseeds are tolerant to Cd stress.

Background: Accumulating documents have demonstrated that long noncoding RNAs (lncRNAs) play critical roles in tumorigenesis. As an lncRNA, nuclear-enriched abundant transcript 1 (NEAT1) has been identified to be involved in the progression of many types of cancers. However, the biological function of NEAT1 in cervical cancer is not fully investigated. The aim of this study was to disclose the specific biological function of lncRNA NEAT1 in cervical cancer progression. Methods: Quantitative real-time polymerase chain reaction (qRT-PCR) was utilized to identify the expression of lncRNA NEAT1 in the cervical cancer tissues and cell lines. All cervical cancer samples used in this study were collected from the Affiliated Suzhou Hospital of Nanjing Medical University between September 2012 and September 2017. The correlation between NEAT1 expression and the overall survival rate of cervical cancer patients was analyzed by Kaplan-Meier analysis. The effects of NEAT1 knockdown or overexpression on cell proliferation were tested by performing MTT assays and colony formation assays. Transwell assays were conducted to detect the migratory ability of cervical cancer cells, in which NEAT1 was silenced or overexpressed. Western blotting was utilized to validate whether NEAT1 promotes cervical cancer progression through activating PI3K-Akt signaling pathway. Results: High expression of NEAT1 predicted poor prognosis of cervical cancer patients (χ2 = 0.735, P = 0.005). Knockdown of NEAT1 decreased the number of colonies in CaSki cell from 136.667 ± 13.503 to 71.667 ± 7.506 (t= −18.76, P = 0.003) and decreased the number of colonies in HeLa cell from 128.667 ± 13.317 to 65.667 ± 7.024 (t = −5.54, P = 0.031). However, overexpression of NEAT1 increased the number of colonies in SiHa cell from 84.667 ± 12.014 to 150.667 ± 18.037 (t = 7.27, P = 0.018). Knockdown of NEAT1 decreased the migratory number of CaSki cell from 100.333 ± 9.866 to 58.333 ± 5.859 (t = −8.08, P = 0.015) and reduced the migratory number in HeLa cell from 123.667 ± 12.097 to 67.667 ± 7.095 (t = −6.03, P = 0.026). Overexpression of NEAT1 increased the migratory number of SiHa cell from 127.333 ± 16.042 to 231.333 ± 31.786 (t = 4.92, P = 0.039). Conclusion: NEAT1 may exert oncogenic function in cervical cancer and serve as a novel therapeutic target for cervical cancer. Key words: Cervical Cancer; Long Noncoding RNA Nuclear-Enriched Abundant Transcript 1; Migration; PI3K/Akt Signaling Pathway; Proliferation

Four previously unreported sesquiterpenoid diasteromers, arteannoides U–X (1–4), together with one new norsesquiterpenoid 5 (arteannoide Y) and one undescribed rearranged cadinene sesquiterpenoid 6 (arteannoide Z) were obtained from the dried aerial parts of Artemisia annua (Qinghao). Notably, arteannoides U–X (1–4) are four stereoisomers that possess the same molecules and the same planar connectivity, but differ from each other in configuration at a certain stereocenter. Their accurate structures were unambiguously identified and distinguished by extensive spectroscopic analyses, NMR calculations with DP4+ analysis, electronic circular dichroism (ECD) calculations and X-ray diffraction analyses. Compounds 1, 3, and 4 showed inhibitory activities against the production of inflammatory cytokines (PGE2, NO, IL-6 and TNF-α) in lipopolysaccharide (LPS)-induced RAW 264.7 macrophages.

Cryoprotective saccharides are widely accepted additives that reduce thawing loss, maintain texture, and retard protein denaturation in the frozen seafood. The present study aimed to investigate the roles of trehalose and alginate oligosaccharides on cryoprotection of frozen shrimp, primarily focusing on the interactions between myosin and saccharide molecules using a molecular dynamics (MD) simulation analysis. The results indicated that soaking in the trehalose and alginate oligosaccharides solutions markedly reduced thawing and cooking losses in frozen shrimp, with respective values decreasing to 6.02%, 8.14%, and 5.99%, 8.19% after 9weeks of storage, which were significantly lower than that of fresh water treatment (9.75% and 15.09%). Our assumption was that water replacement played a leading role in cryoprotection, as shown in previous experimental results and reports. Furthermore, homology modeling and MD simulations confirmed that trehalose and alginate oligosaccharides substituted the water molecules around the myosin surface by forming hydrogen bonds with polar residues of amino acids, thereby stabilizing the structures in the absence of water during frozen storage. These conditions affected the flexibility of particular amino acid residues, enhanced the residue cross correlations within the two chains of myosin, and also increased the total interaction energy between myosin and water/saccharide molecules, thereby leading to an increase in protein stability. Finally, by comparing the experimental results to that of MD simulation, significant positive correlation existed between saccharides and the stabilization of myosin in shrimp muscle. The findings of the present study may help better understand the cryoprotective mechanisms of saccharides in frozen shrimp, and the two saccharides may be potentially used as alternative additives in seafood to maintain better quality during frozen storage.

Two new iridoid glycosides, 2′-O-cis-coumaroylgardoside (1), and 6′-O-caffeoylioxide (2), were isolated from the fruit of Gardenia jasminoides. The structures of these compounds were elucidated based on spectroscopic analysis (HR-ESI-MS, NMR) and chemical methods. The anti-inflammatory activities of the isolates were evaluated by measuring their inhibitory effects on PGE2 production in LPS stimulated RAW 264.7 macrophages, compounds 1 and 2 could reduce PGE2 levels in LPS-activated RAW 264.7 macrophages with IC50 values of 121.4 and 83.38 μM, respectively.

New vadimezan hydrazones were synthesized as E/Z isomers and characterized by IR, 1H NMR, MS, and HRMS techniques; the ratio of isomers was analyzed by NOESY spectra. All compounds were evaluated for their cytotoxic activities against endometrial Ishikawa, lung A549 and NCI-460, chorion Bewo, cervical HeLa and Siha, breast MCF-7, leukemia HL-60, liver BEL-7402, and stomach BGC-823 cancer cell lines in vitro by standard MTT assay. The preliminary pharmacological results indicated that compound 5c exhibited most promising cytotoxic activity against NCI-460 cell line with IC50 value of 7.01 ± 0.53 μM.

Thirteen new sesquiterpenoids, arteannoides F-R (1-13), along with 13 known analogues (14-26), were isolated from the dried aerial parts of Artemisia annua L. Their structures, including absolute configurations, were unambiguously determined by a combination of physical data analyses (HRESIMS, 1D and 2D NMR, and ECD) as well as the crystal structures of 1, 5, 6, 15, 19, and 23. Among the isolated compounds, 1 features an unusual 11-oxatricyclo[6.2.1.04,9]undecan-2-ene ring system, 5 possesses an uncommon 4,11-ether bridged tricyclic framework, whereas 6 is a new eudesmane-type sesquiterpenoid formed via rearrangement of its carbon backbone. The systemically anti-inflammatory activities of all isolates were evaluated by measuring their inhibitory effects on PGE2, NO, TNF-α, and IL-6 production in LPS-stimulated RAW 264.7 macrophages. Moreover, the structure activity relationships of some compounds are summarized, this study will provide new structural templates for discovering potential anti-inflammatory agents.

F-box protein is a subunit of Skp1-Rbx1-Cul1-F-box protein (SCF) complex with typically conserved F-box motifs of approximately 40 amino acids and is one of the largest protein families in eukaryotes. F-box proteins play critical roles in selective and specific protein degradation through the 26S proteasome. In this study, we bioinformatically identified 972 putative F-box proteins from Medicago truncatula genome. Our analysis showed that in addition to the conserved motif, the F-box proteins have several other functional domains in their C-terminal regions (e.g., LRRs, Kelch, FBA, and PP2), some of which were found to be M. truncatula species-specific. By phylogenetic analysis of the F-box motifs, these proteins can be classified into three major families, and each family can be further grouped into more subgroups. Analysis of the genomic distribution of F-box genes on M. truncatula chromosomes revealed that the evolutional expansion of F-box genes in M. truncatula was probably due to localized gene duplications. To investigate the possible response of the F-box genes to abiotic stresses, both publicly available and customer-prepared microarrays were analyzed. Most of the F-box protein genes can be responding to salt and heavy metal stresses. Real-time PCR analysis confirmed that some of the F-box protein genes containing heat, drought, salicylic acid, and abscisic acid responsive cis-elements were able to respond to the abiotic stresses.

Arteannoide A (1), an unusual cadinane dimer featuring a rare fused 6,8-dioxabicyclo[3.2.l]octan-7-one ring system, arteannoides B and C (2 and 3), two novel heterodimers incorporating a rearranged cadinene sesquiterpenoid and a phenylpropanoid, together with two new rearranged cadinene sesquiterpenoids 4 and 5, were isolated from Artemisia annua L. Their structures were determined by a combination of NMR spectroscopy, electronic circular dichroism calculations, and X-ray diffraction analyses. Compounds 2 and 3 showed inhibition of nitric oxide production in lipopolysaccharide-induced RAW 264.7 mouse macrophage cell lines with IC50 values of 4.5 and 2.9 μM, respectively.

A new dimeric secoiridoids derivative, named japonicaside E, together with six known compounds were isolated from the flower buds of Lonicera japonica. The structures of these compounds were elucidated by the analyses of mass spectrometry and NMR spectroscopy. All compounds were evaluated for anti-inflammatory activity in vitro.

Two new labdane diterpenoids, Leojaponin E (1) and F (2), together with three known compounds were isolated from the dried herb of Leonurus japonicus Houtt., Lamiaceae. Their structures were determined based on extensive spectroscopic analyses. The absolute configurations of 1 and 2 were elucidated on the basis of experimental and calculated electronic circular dichroism spectra. In addition, compounds 1 and 2 exerted inhibition of LPS-induced PGE2 production in a dose-dependent manner at concentrations ranging from 5 to 20 μM.

Keywords: NMR; 1H NMR; 13C NMR; 2D NMR; Compositae; Artemisia annua; sesquiterpenoid

To evaluate the protective effects of Reduning Injection (热毒宁注射液, RDN), a patent Chinese medicine, on lipopolysaccharide (LPS)-induced acute lung injury (ALI) in rats and its underlying mechanisms of action. Sixty male Sprague-Dawley rats were randomly divided into 6 groups, including normal control, model, dexamethasone (DEX, 5 mg/kg), RDN-H (720 mg/kg), RDN-M (360 mg/kg) and RDN-L (180 mg/kg) groups, with 10 rats in each group. Rats were challenged with intravenous injection of LPS 1 h after intraperitoneal treatment with RDN or DEX. At 6 h after LPS challenge, lung tissues and bronchoalveolar lavage fluid (BALF) were collected, and the number of inflammatory cells was determined. The right lungs were collected for histopathologic examination, measurement of gene and protein expressions, superoxide dismutase (SOD) and myeloperoxidase (MPO) activities. In vivo pretreatment of RDN (360, 720 mg/kg) significantly reduced the weight of wet to dry (W/D) ratio of lung, protein content in BALF, and led to remarkable attenuation of LPS-induced histopathological changes in the lungs. Meanwhile, RDN enormously decreased BALF total inflammatory cells, especially neutrophil and macrophage cell numbers. Moreover, RDN increased SOD activity, inhibited MPO activity, alleviated LPS-induced tumor neurosis factor-α (TNF-α) and inducible nitric oxide synthase (iNOS) expression in lung tissues. Furthermore, RDN (720 mg/kg) efficiently weakened nuclear factorkappa B (NF-κB) gene and protein expression. Anti-inflammatory effects of RDN was demonstrated to be preventing pulmonary neutrophil infiltration, lowering MPO activity, TNF-α and iNOS gene expression by inhibiting NF-κB activity in LPS-induced ALI.

A 3 factor, 3 level Box-Behnken factorial design combined with response surface methodology was used to optimize the fermentation process for production of extracellular polysaccharide (EPS) from Hirsutella sinensis. The 3 independent variables were temperature (X1), initial pH (X2), and the glucose concentration (X3). Experimental data were fitted to a 2nd order polynomial equation using multiple regression. The optimal fermentation conditions of the production of EPS were a temperature of 18.21°C, an initial pH of 5.81, and a glucose concentration of 7.39 g/L. The maximum predicted EPS production of 2.41 g/L was close to the actual experimental EPS yield (2.42±0.038%), demonstrating the model validity. EPS from H. sinensis had a direct in vitro immuno-stimulating activity using murine macrophage RAW264.7 cells, and stimulated release of several major cytokines (IL-1β, NF-κB, TNF-α, and iNOS) in a dose-dependent manner.

Ethnopharmacological relevance Cuscuta chinensis ( C. chinensis ) is a well-known traditional Chinese herb that has been used to treat heart disease, diabetes, liver injury, cancer, and aging. Materials and methods Murine osteoblastic MC3T3-E1 cells were treated with various concentrations of C. chinensis water extraction at different time intervals. The antioxidant effect of C. chinensis on MC3T3-E1 cells was evaluated using MTT and TUNEL assays. The effect of C. chinensis on cell cycle was analyzed by flow cytometry with propidium iodide. Lipid peroxidation was measured by the HPLC method. The cellular redox status was determined from the reduced glutathione to oxidized glutathione ratio (GSH/GSSG) and the enzymes involved in glutathione metabolism, including glutathione reductase (GR), Glutathione S-transferase (GST), and Glucose-6-phosphate dehydrogenase (G6PD). The changes in relative mitochondrial transmembrane potential (Δ Ψm ) in the MC3T3-E1 cells were analyzed with rhodamine 123 staining. Western blot analysis was used to evaluate the levels of cytochrome c (cyto c ), Bax, Bcl-2, caspase 3, Sirt3, and IDH2 expressions. Results The C. chinensis water extraction protects tertiary butyl hydroperoxide (TBHP)-treated MC3T3-E1 cells from death in a dose-dependent manner. C. chinensis treatment significantly inhibited the reactive oxygen species (ROS) generation, malondialdehyde (MDA) production, and increased the activity of superoxide dismutase (SOD), GR, GST, and G6PD. The release of cyto c from mitochondria was reduced by C. chinensis , which increased the expression of antiapoptotic IDH2, Sirt3, and Bcl-2 and decreased the expression of Bax, cyto c , and caspase 3. Conclusions C. chinensis modulated the oxidative stress-induced apoptosis in MC3T3-E1 cells, probably due to its antioxidant activity and functioning via mitochondria-dependent pathways.

Strike a pose! A series of 4-anilinoquinazolines were designed, synthesized and evaluated in vitro against lung and breast cancer cell lines. Several compounds were found to be endowed with cytotoxicity in the low micromolar range. Molecular docking suggests that these compounds bind to EGFR in a similar manner to known EGFR inhibitors.

Casuarina species are valued as pioneer trees for coastal shelter forest construction. However, low temperature stress limits its cultivation expansion, fast growth, and high yield. To investigate the molecular mechanism by which Casuarina trees cope with cold stress and to provide a comprehensive sequence resource for further molecular genetic research on Casuarina species, we employed RNA-Seq technology to study the genome-wide expression profiles during cold stress in Casuarina equisetifolia L. Four cDNA libraries constructed from mRNAs of control and cold-treated seedlings (at −5 °C for 5 h) were sequenced by using the Illumina sequencing platform. A total of 118,270 unigenes were obtained by de novo assembly, creating an initial reference transcriptome dataset of C. equisetifolia. Among these unigenes, 4639 were identified as early cold response genes including 2331 cold up-regulated and 2308 down-regulated. Furthermore, the expression changes of 18 genes under cold stress from RNA-Seq analysis were further validated by reverse transcription quantitative polymerase chain reaction (RT-qPCR). Functional network analysis indicate that modifications in the expression of genes involved in signal regulation by transcription factors, metabolic pathways including synthesis of carbohydrate, secondary metabolites, and antioxidant enzymes might play vital roles in responses to cold stress in C. equisetifolia. This study represents the first large-scale transcriptome annotation of C. equisetifolia and may be beneficial to elucidate the cold response mechanisms of Casuarina trees and will be useful in cold-tolerant Casuarina species breeding.

A rare hetero dimeric terpenoid derivative, named japonicaside C, together with five known secoiridoid gloucosides were isolated from the flower buds of Lonicera japonica. The structures of these compounds were established on the basis of spectroscopic analyses. Japonicaside C is the first representative of a novel type of hetero dimeric terpenoid, biogenetically originated from a guaiane-type sesquiterpenoid and a secoiridoid glucoside. Anti-virus activity of the isolated compounds was evaluated in vitro.

Xyloglucan endotransglucosylase/hydrolases (XTH) are one of the key enzymes regulating cell wall construction, extension and metabolism. In the study, 44 XTH protein genes from Medicago truncatula genome were identified using bioinformatics, microarray and RT-PCR. Each XTH was showed to possess a highly conserved domain ((D/N)-E-(I/L/F/V)-D-(F/I/L)-E-(F/L)-L-G), and most of XTHs possess four Cys in the C terminal region, which suggests the potential for generating disulfide bonds. Based on the XTH protein sequences, these XTHscan be classified into three major families and each family can be subdivided into more groups. Examination of the genomic location of XTH genes on M. truncatula chromosomes showed that the evolutional expansion of the genes was possibly attributed to localized gene duplications. To investigate the possible involvement of the XTHs responding to heavy metals and other abiotic stresses, the XTH genes were exposed to heavy metal (Hg or Cu), salt and drought stresses. There were 28, 21 and 21 MtXTH genes found to respond to HgCl2, salt and drought stresses, respectively, but their expression were different under the stresses. Some of the XTH genes were well confirmed by quantitative RT-PCR (qRT-PCR). We further specified expression of a XTH gene Medtr4g128580 (MtXTH3) under different environmental stresses, and showed that MtXTH3 was induced by Hg exposure. These results indicated that a group of MtXTHs could be differentially expressed under the environmental stresses.

The aim of this article is to investigate the mechanism of vitexin-4''-O-glucoside (VOG) protecting ECV-304 cells against tertbutyl hydroperoxide (TBHP)-induced injury. ECV-304 cell viability was measured by MTT assay. Apoptosis was determined by terminal deoxynucleotidyl transferase-mediated dUTP nick end-labelling (TUNEL) assay. Cellular morphological changes were observed using phase contrast microscopy. The change of relative mitochondrial transmembrane potential in the ECV-304 cells was analysed with rhodamine 123 staining. Lipid peroxidation was measured by the HPLC method. The results showed that 128 µmol L(-1) VOG could effectively protect ECV-304 cells against cytotoxicity induced by TBHP. VOG protected TBHP-treated ECV-304 cells from death, significantly decreased MDA production, and increased superoxide dismutase (SOD) activity and mitochondrial membrane potential (ΔΨ). Taken together, VOG protects against TBHP-induced ECV-304 cell injury partially through resuming mitochondrial function.

Aim Accumulating evidence has indicated that Panax notoginseng saponins (PNS), the major ingredients in Panax notoginseng ( Burk. ) F.H. Chen which could be found widely in Asia, can attenuate atherogenesis in vivo . This study was designed to examine the relationship of PNS with cholesterol ester in foam cells sourced from macrophages and the effect of PNS on the expression of ATP-binding cassette transporter A1 (ABCA1). Materials and methods Foam cells sourced from macrophages were cultured with PNS. The content of cholesterol ester in foam cells was analyzed and expressions of ABCA1 and liver X receptor α (LXRα) in foam cells were measured by real-time PCR and western blotting methods. Results The results showed that PNS could significantly decrease the level of cholesterol ester in foam cells at middle and high dosages. The real-time PCR and western blotting assays indicated that the expression of ABCA1 was up-regulated by PNS in a dose-dependent manner. Analysis based on these results showed that the cholesterol ester level was negatively correlated with ABCA1 expression. Conclusions As a result, we conclude that by up-regulating the expression of ABCA1, PNS could lower the cholesterol ester level, which resulted in the attenuation of the foam cell formation. This bioactivity might be associated with the special chemical structures of PNS that are similar to the natural agonist of LXRα.

A simple high-performance liquid chromatographic (HPLC) assay using the internal standard method is developed for the simultaneous determination of eight polyphenols. The analyzed compounds isolated from the leaves of Crataegus pinnatifida Bge. var. major include chlorogenic acid, vitexin-4"-O-glucoside, vitexin-2"-O-rhamnoside, vitexin, rutin, hyperoside, isoquercitrin, and quercetin. HPLC analysis is performed on a Diamonsil C18 analytical column (150 x 4.6 mm, i.d., 5-microm) using solvent (A) acetonitrile-tetrahydrofuran (95:5, v/v) and (B) 1% aqueous phosphoric acid as the mobile phase with UV absorption at 270 nm. The calibration curves of the eight polyphenols are linear (r(2) > 0.9992) over the concentration range of 0.0894-120.0 microg/mL. The mean recoveries are 95.4% to 98.1%. The results indicate that the HPLC method developed can easily be applied to the determination of eight polyphenols in the leaves of Crataegus pinnatifida Bge. var. major.

Forty caffeate analogues were synthesized via a convenient method starting from vanillin with moderate to good yields. The testing of biological activity of these compounds against HIV-1 integrase indicates that four compounds: bornyl caffeate, bornyl 2-nitrocaffeate, 5-nitrocaffeic acid and 5-nitrocaffeic acid phenethyl ester (5-nitroCAPE) possess a good HIV integrase inhibitory activity, IC(50) 19.9, 26.8, 25.0 and 13.5 microM , respectively. Twelve caffeate analogues were tested by MTT assay on growth of human hepatocellular carcinoma BEL-7404, human breast MCF-7 adenocarcinoma, human lung A549 adenocarcinoma and human gastric cancer BCG823 cell lines, respectively. And the best result is IC(50) 5.5 microM for CAPE against BEL-7404.

To investigate the antioxidant effect of vitexin-4''-O-glucoside, a flavone glycoside, isolated from the leaves of Crataegus pinnatifida Bge. var. major, we developed a simple and sensitive high-performance liquid chromatography (HPLC) method to determine levels of malondialdehyde (MDA) in ECV304 cell culture medium after induction by tert-butyl-hydroperoxide (TBHP). The preparation of analyzed samples involved a one-step derivatization with thiobarbituric acid (TBA). HPLC analysis was performed on a Synergi Hydro-RP, a polar end-capped C18 column (250 x 4.6 mm, 4 mum), using an acetonitrile-ammonium acetate aqueous solution (10 mM, pH 6.8) as the mobile phase under linear gradient conditions with UV detection at 532 nm. The calibration curve was linear over 0.0125-1.25 microM MDA (r = 0.9951). Relative standard deviations (RSDs) of intra-day and inter-day precision were less than 6.1% and 5.0%, respectively. The mean recovery was 96.9 +/- 1.6%. The lower limit of quantification (LLOQ) of MDA was 0.0125 microM. This chromatographic method was successfully applied to investigating the in vitro antioxidant effect of vitexin-4''-O-glucoside. Vitexin-4''-O-glucoside (120 M) protected ECV304 cells from peroxidation induced by TBHP.

We have reported that the protective effect of Magnolol on TBHP-induced injury in human nonsmall lung cancer H460 cells is partially via a p53 dependent mechanism. In this study, we found that Magnolol displayed a stimulatory effect at low concentrations (or = 20 microM) whilst inhibitory effect at high concentrations (or = 40 microM) in H460 cells. To investigate the mechanism of inducing the biphasic effect in H460 cells with Magnolol, we showed that Magnolol stimulated DNA synthesis at low concentrations and displayed an inhibition effect at high concentrations in H460 cells. More importantly, the inhibition of DNA synthesis was accompanied by the S phase cell cycle arrest and the appearance of intense intracytoplasmic vacuoles. These vacuoles can be labeled by autophagic marker monodansylcadaverin (MDC), 3-methyladenine (3-MA), an inhibitor of autophagy, was able to inhibit the occurrence of autophagy. The results of the LDH activity assay and TUNEL assay also showed that Magnolol at high concentrations inhibiting H460 cell growth was not via apoptotic pathway. Furthermore, accompanied by the occurrence of autophagy, the expression of phospho-Akt was down-regulated but PTEN significantly was up-regulated. In conclusion, Magnolol induces H460 cells death by autophagy but not apoptotic pathway. Blockade of PI3K/PTEN/Akt pathway is maybe related to Magnolol-induced autophagy. Autophagic cells death induction by Magnolol underlines the potential utility of its induction as a new cancer treatment modality.

To study the metabolic transformation of pumiloside by rat intestinal flora in vitro and identify its metabolites. Pumiloside was incubated in the rat intestinal flora in vitro. HPLC was used to monitor the metabolic process, and HPLC-Q-TOF-MS was used to identify the structures of biotransformation products. In vitro, pumiloside was easily metabolized by rat intestinal flora, and with the prolongation of metabolic time, pumiloside was transformed into several metabolites. Three metabolites were initially identified in this experiment. The study indicated that pumiloside could be extensively metabolized in the rat intestinal flora in vitro.

Problem The relationship between genital infection and two sperm parameters, namely, concentrations of nitric oxide (NO) and cytokines [e.g., interleukin (IL)-17 and IL-18], in the semen of infertile males remains undetermined. Method of study Semen samples from 81 infertile (with or without Ureaplasma urealyticum infection) and normal males were subjected to semen analysis. Results Nitric oxide concentration in the semen of infertile males with genital infection was higher than those of infertile males without genital infection and of normal males (P < 0.05). Sperm density, pH, percentage of forward, movement of sperm, sperm activation rate, sperm survival rate, and normal rate of the sperm morphology of infertile males with U. urealyticum infection were significantly lower than those of infertile males without genital infection and of normal males (P < 0.05). NO concentrations were also positively correlated with IL-17 and IL-18 concentrations in the semen of infertile males. Conclusion Increased NO concentration and abnormal IL-17 and IL-18 expression were induced by genital infection and induced damage to male reproductive capacity, thereby causing male infertility.

Basidiomycete species accumulate high levels of heavy metals, particularly cadmium (Cd). Heavy metal toxicity has been widely investigated, and different key factors affecting tolerance to heavy metals have been identified in plants and fungi. Nevertheless, molecular insights into different mechanisms of fungi to directly resist reactions with particular heavy metals remain insufficient. As such, early-response genes in basidiomycete fungi must be identified to elucidate the mechanisms that confer resistance to heavy metal stress, particularly for Cd. The basidiomycete fungus Lentinula edodes (Berk.) Pegler, an edible mushroom (shiitake), is the most economically important cultivated mushroom. Suppression subtractive hybridization (SSH) combined with mirror orientation selection (MOS) was performed to identify Cd-induced genes in L. edodes treated with 50 μM Cd for 24 h. A total of 72 L. edodes genes were identified from the forward subtractive library. Reverse transcription quantitative polymerase chain reaction (RT-qPCR) was then employed to confirm the induction of the identified genes upon Cd exposure. Bioinformatics analysis revealed that 53 of the identified genes encode proteins with known functions and may be involved in early response to Cd exposure, whereas 19 genes encode hypothetical proteins with unknown functions. The diversity of the putative functions of the identified genes indicates that Cd stress elicits complex responses in L. edodes, and complex metabolic, transport, and signaling pathways may be involved in Cd resistance. This study presents a group of potential early-response genes to Cd exposure and may be beneficial to elucidate heavy metal resistance in basidiomycete fungi.

Three new iridoid glucosides, 6″-O-trans-feruloylgenipin gentiobioside (1), 2'-O-trans-caffeoylgardoside (2), jasmigeniposide A (3), and one new bis-iridoid glucoside, jasmigeniposide B (4), along with six known analogues (5-10), were isolated from the fruit of Gardenia jasminoides. Their structures were established on the basis of spectroscopic analyses and chemical methods. Anti-virus activity of isolated compounds were evaluated in vitro and only compound 9 showed moderate inhibitory activity against H1N1 with 50% effective concentration (EC50) value of 104.36 μM, and selective index (SI) value greater than 4.79.

Ligularia is mainly distributed in the western regions of China. Most of the species have been traditionally used in folk medicine for the treatment of hepatitis B, asthma, hemoptysis and pulmonary tuberculosis. In our continuation of research on antiviral components from traditional Chinese medicine, Ligularia atroviolacea was tested for inhibitory effects on hepatitis B virus (HBV). A bioassay-guided phytochemical examination of L. atroviolacea disclosed that its ethyl acetate extract, which was made up of two eremophilenolides, showed suppressive activity on the expression of HBV surface antigen (HBsAg) in the HepG2.2.15 cell line. Then a simple and effective preparative high-speed counter-current chromatography method was successfully developed for the isolation and purification of two main active metabolites, 8beta-hydroxyeremophil-3,7(11)-dien-12,8alpha;15,6alpha-diolide and 8beta-methoxyeremophil-3,7(11)-dien-12,8alpha;15,6alpha-diolide from the ethyl acetate extract of L. atroviolacea by a one-step separation using a two-phase solvent system composed of light petroleum (60-90 degrees C)-ethyl acetate-methanol-water (9:1:8:2, v/v/v). The chemical structures of the two eremophilenolides were identified by ESI-MS, (1)H-NMR and (13)C-NMR analysis. The anti-HBV activity of the two purified compounds was measured; both of them showed suppressive activity on the expression of HBsAg in the HepG2.2.15 cell line. The results support the continued and expanded exploitation and utilization of L. atroviolacea.

The aim is to investigate the effect of Magnolol preserved H460 cells from an oxidative agent tert-butylhydroperoxide (TBHP)-induced cell death. Magnolol augmented cell survival ratio after TBHP challenged. The protective action of this drug was more efficacious than that of N-acetylcysteine (NAC) which is a putative antioxidant. DNA damage, detected by the comet assay, was diminished after treatment of Magnolol. The cells viability decreased after treatment with 0.15 mM TBHP for 24 h, accompanied by inducing apoptotic death of the cells. Cytotoxicity and apoptosis induced by TBHP were significantly inhibited or attenuated after pretreatment with 20 microM Magnolol. Magnolol contributes to the cells survival through downregulated the p53 phosphorylation and PTEN expression, and upregulated Akt phosphorylation. Taken together, Magnolol was effective against DNA single strand breaks (SSB) formation, cytotoxicity and lipid peroxidation induced by TBHP, and its effects on p53 phosphorylation, PTEN and Akt phosphorylation were due to its antioxidative function, and partially via a p53 dependent mechanism in this protective effects.