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Graphical abstract, Introduction Previously, we have reported a withanolide-type steroid, named tubocapsenolide A (TA), which shows potent anti-proliferative activity in several cancer cell lines. However, its inhibitory effect on the Janus kinase/signal transducer and activator of transcription 3 (JAK/STAT3) pathway and therapeutic potential on osteosarcoma have not been reported. Objectives In the present study, we aimed to investigate the effect and molecular mechanism of TA in osteosarcoma. Methods The biological functions of TA in U2OS cells were investigated using colony formation, 5-ethynyl-20-deoxyuridine (EDU) staining, and cell cycle/apoptosis assays. The interaction between TA and Src homology 2 phosphatase 2 (SHP-2) was detected by enzyme activity and validated by target-identification methods such as drug affinity responsive target stability (DARTS), cellular thermal shift assay (CETSA), and biolayer interferometry (BLI). The in vivo anti-tumor efficacy of TA was analyzed in the xenograft tumor model. Western blotting analysis was performed to detect the protein expression levels. Results TA exhibited antitumor activity against osteosarcoma both in vitro and in vivo by regulating the JAK/STAT3 signaling pathway. Mechanically, TA interacted with SHP-2 directly and activated its phosphatase activity. Importantly, protein tyrosine phosphatase (PTP) inhibitor, SHP-2 inhibitor, and SHP-2 siRNA could reverse the inhibitory effect of TA on the JAK/STAT3 signaling pathway and restored the TA-induced cell death. Conclusion TA activated the phosphatase activity of SHP-2, which resulted in the inhibition of the JAK/STAT3 pathway and contributed to the antitumor efficacy of TA. Collectively, these findings suggested that TA could serve as a novel therapeutic agent for the treatment of osteosarcoma.

Six new oligomeric neolignans including two trimeric neolignans (1 and 2) and four dimeric neolignans (3-6) were isolated from the leaves of Magnolia officinalis var. biloba. Their structures were determined based on HR-ESIMS and NMR data, as well as electronic circular dichroism (ECD) calculations. Compound 1 is formed from two obovatol moieties directly linked to an aromatic ring of the remaining obovatol moiety, which is an unprecedented type of linkage between monomers. All isolates were assessed for their inhibitory effects on NO production in LPS-stimulated RAW 264.7 macrophage cells. Compounds 1 and 3 showed significantly inhibitory activities with IC50 values of 6.04 and 3.26 μmol·L-1, respectively.

Four new highly oxidized sesquiterpene lactones, Argyoxynolides A–D, along with two known analogs were isolated from the leaves of Artemisia argyi H.Lev. & Vaniot. Their structures were characterized utilizing extensive spectroscopic methods. All isolates were evaluated for the LPS-induced nitric oxide (NO) production inhibitory activity and one compound exhibited activity with IC50 = 37.95 ± 4.39 μM.

On-demand therapy following effective tumor detection would considerably reduce the side effects of traditional chemotherapy. DT-diaphorase (DTD), whose level is strongly elevated in various tumors, is a cytosolic flavoenzyme that promotes intracellular reactive oxygen species (ROS) generation via the redox cycling of hydroquinones. Incorporation of the DTD-responsive substrate to the structures of the probe and prodrug may facilitate the tumor detection and therapy. Herein, we established an multifunctional drug delivery nanosystem (HTLAC) that rapidly responds to the DTD enzyme, leads to the early-stage precise detection and termination of tumors. Firstly, the synthesis of DTD-responsive withaferin A (DT-WA) and indocyanine green (DT-Cy5) was performed. In the presence of DTD, WA, which produces ROS in cells, was released from DT-WA, and the red fluorescence of DT-Cy5 was detected for tumor imaging. Additionally, these DTD enzyme reaction processes of DT-WA and DT-Cy5 induced ROS. The self-burst of ROS generation by the two enzyme reaction processes as well as the released WA then led to the apoptosis of tumor cells. To increase the bioavailability and tumor targeting of drugs, cell-penetrating peptide and hyaluronic acid functionalized liposomes were used to encapsulate the drugs. The detailed in vitro and in vivo assays showed that HTLAC achieved enhanced tumor detection and superior antitumor efficiency. According to above outcomes, results showed that HTLAC might provide an efficacious approach for the fabrication of enzyme-triggering nanosystems to detect tumor and induce the self-burst of ROS for an efficient tumor treatment. STATEMENT OF SIGNIFICANCE: We have fabricated a HTLAC nanosystem to address the need of bursting reactive oxygen species (ROS) generation within tumor site. Our goal uniquely aims at not only augmentation of ROS-inducing anticancer efficacy, but also to meet the challenges of tumor dynamic detection in the clinical practices. In this work, the DT-diaphorase responsive withaferin A (DT-WA) and indocyanine green (DT-Cy5) are synthesized, and observed more specifically toward DTD under physiological conditions. As the cell-penetrating peptide and hyaluronic acid functionalized liposome, the HTLAC not only induces antiproliferative activity by generating self-burst of ROS, but also effectively accumulate and restore its fluorescence at the tumor site because of the HA actively targeting tumor along with the prolonged presence in blood circulation. Besides, this enzyme-triggering nanosystem exhibited an effective tumor inhibition with a low systemic toxicity.

Nonalcoholic steatohepatitis (NASH) is a common clinical condition that can lead to advanced liver diseases. The mechanism of the diaease progression, which is lacking effective therapy, remains obsure. Therefore, there is a need to understand the pathogenic mechanisms responsible for disease development and progression in order to develop innovative therapies. To accomplish this goal, experimental animal models that recapitulate the human disease are necessary. Currently, an increasing number of studies have focused on natural constituents from medicinal plants which have been emerged as a new hope for NASH. This review summarized the pathogenesis of NASH, animal models commonly used, and the promising targets for therapeutics. We also reviewed the natural constituents as potential NASH therapeutic agents.

Artemisianins A-D (1–4), four stereoisomers of sesquiterpenoid dimers, forming via [4+2] cycloaddition from a 1, 10-seco-guaianolide dienophile and a guaianolide diene, along with two biosynthetically related precurors 5 and 6, were isolated from the famous traditional Chinese medicine Artemisia argyi. The structures of 1–4, including their absolute configurations, were elucidated by extensive spectroscopic data and ECD/TDDFT calculation analysis. Compounds 1–4 exhibited cytotoxicity with IC50 values ranging from 7.2 to 23.3 μM. The accumulation of Ca2+ in cytoplasm and enlarged endoplasmic reticulum (ER) indicated that 1 mediated HT-29 cancer cell apoptosis through improvement of ER-stress, which was further proved by unfolded protein response (UPR) pathway on basis of the upregulation of IRE1α, p-PERK, ATF6, and CHOP.

Five new prenylated acetophenones, melicoptelins A-E (1–5), along with one known congener (6) were isolated from the roots of Melicope ptelefolia. Among them, compounds 2a/2b, 3a/3b, and 4a/4b were obtained as inseparable interconverting mixtures of keto and enol tautomers. Their structures were elucidated on the basis of extensive spectroscopic methods, including 1D, 2D NMR and HRESIMS. Compouds 2a/2b, 4a/4b and 5 exhibit protein tyrosine phosphatase 1B (PTP1B) inhibitory activity with IC50 values of 34.4, 55.2 and 66.6 μM, respectively.

Synephrine, a chiral adrenergic agonist, exists as R- (more active) and S-enantiomers (less active) that are difficult to separate because of their alkalinity and high polarity. Herein, we show that although this enantioseparation can hardly be achieved by high-performance liquid chromatography, acceptable preparative separation can be realized using high-speed counter-current chromatography (HSCCC) and pH-zone-refining counter-current chromatography (CCC) under optimal conditions. Specifically, 23.0 mg of S-synephrine and 25.0 mg of R-synephrine were isolated from 60.0 mg of racemic synephrine by conventional HSCCC, while 70.0 mg of S-synephrine and 69.0 mg of R-synephrine were obtained from 200.0 mg of racemic synephrine by pH-zone-refining CCC. The pH-zone-refining CCC was identified as the most efficient enantioseparation technique for synephrine and can be applied for the preparative enantioseparation of other β-amino alcohols.

Effective targeting of mitochondria has emerged as a beneficial strategy in cancer therapy. However, the development of mitochondria-targeting ligands is difficult because of the low permeability of the mitochondrial double membrane. We found that hypericin (HY), a natural product isolated from Hypericum perforatum L., is an effective mitochondria-targeting ligand. HY-functionalized graphene oxide (GO) loaded with doxorubicin (GO-PEG-SS-HY/DOX) increased the synergistic anticancer efficacy of phototherapy and chemotherapy in the absence of apparent adverse side effects. In vitro and in vivo assays suggested GO-PEG-SS-HY/DOX induced the expression of the key proteins of the mitochondria-mediated apoptosis pathway and caused apoptosis of breast carcinoma cells. In addition, GO vehicle exhibited low toxicity toward normal cells, indicating high safety of functionalized GO preparations in antitumor therapy. Therefore, HY-functionalized GO can be successfully used as a platform technology to target mitochondria in cancer cells and improve the therapeutic efficacy of chemotherapeutic drugs. Statement of Significance Induction of mitochondria-mediated apoptosis is a promising approach in cancer therapy. However, mitochondria are difficult to access and permeate because of their negative membrane potential and highly dense double membrane. Mitochondria-targeting ligands can be conjugated to nanoparticles or small-molecule drugs to enhance their antitumor effect. Here, we showed that the natural photosensitizer hypericin is a novel mitochondria-targeting ligand and that graphene oxide particles co-loaded with hypericin and the chemotherapeutic agent doxorubicin exhibited a synergistic antitumor effect mediated by the mitochondrial-mediated apoptosis. Treatment with such particles in combination with laser irradiation led to apoptosis of the tumor MDA-MB-231 and MCF-7 cells in vitro and in vivo. Furthermore, treatment with hypericin/doxorubicin-functionalized graphene oxide had low cellular toxicity.

The reactivity-based screening (RBS) was developed for directed discovery of cytotoxic withanolides. In this study, a thiol probe, 4-chlorobenzenethiol, was used to selectively attack cytotoxic withanolides containing potential pharmacophore, 2(3)-en-1-one in ring A (AEO) and 5β,6β-epoxy in ring B (BE), from the plant extract of Physalis angulata var. villosa. The screening was performed based on the potential mechanism of 4-chlorobenzenethiol nucleophilic addition to AEO, followed by detection of adducts using liquid chromatography quadrupole-time-of-flight mass spectrometry (LC-Q-TOF-MS). Guided by RBS, eleven target withanolides, including five new compounds, physagulides R-V (10–14) and six known ones (2, 7–9, 15, 16) were discovered. All of them exhibited cytotoxicity against the both tested cell lines, especially, compounds 2, 7, 8 and 14 showed potent activities with IC50 values of 1.57–6.29 μM. The results suggested that RBS was efficient and accurate for rapid identification of cytotoxic withanolides and could guide isolation of target components from the complex medicinal plant extract.

Seven new phloroglucinol derivatives, uraliones L-R (1-7), were isolated from the whole plant of Hypericum urialum. The structures of new compounds were unambiguously elucidated by comprehensive spectroscopic analysis, and the absolute configurations of the vic-diol groups were determined by Mo2(OAc)4-induced electronic circular dichroism experiments. Their capability to reversal the multidrug resistance in doxorubicin-induced resistant MCF-7/DOX sublines showed that compounds 1, 5, 6, 7 exerted a potential effect of doxorubicin susceptibility.

Seven sucrose esters, physakengoses K-Q ( 1 – 7 ) were isolated from the aerial parts of Physalis alkekengi var. franchetii . Their structures were elucidated on the basis of extensive spectroscopic analyses and chemical methods. These new compounds were tested for their antimicrobial abilities against Staphylococcus aureus , Bacillus subtilis , Pseudomonas aeruginosa and Escherichia coli . Among the isolated sucrose esters, compounds 1 – 5 showed potent antibacterial activity with MIC values ranging from 2.16 to 12.76 μ g/mL.

Six novel calyxins, named calyxin T-W, ent-calyxin T and ent-calyxin U were isolated from the seeds of Alpinia katsumadai Hayata. Their relative configurations were elucidated by means of detailed UV, IR, NMR and MS spectroscopic data. Their absolute configurations were assigned by collaborative studies on single crystal X-ray diffraction analysis, Mosher's method, electronic circular dichroism (ECD), optical rotation and theoretical calculations. These compounds are Friedel-Cranft alkylation adducts composed of coexisted diarylheptanoids and flavanone from the seeds of Alpinia katsumadai. The antiproliferative activity of the six compounds against NCI-H460, HeLa, SMMC-7721 and HCT-116 cell lines was also reported, and most of them showed moderate to strong activities.

A pair of new tetranorlabdane diterpenoid epimers, named elettarins A (1) and B (2), together with five known labdane diterpenes (3–7), were isolated from Elettaria cardamomum Maton. The structures of elettarins A and B were established based on spectroscopic data (HRESIMS, 1D/2D NMR), and ECD experimentation. All isolates were evaluated for their inhibitory activities against nitric oxide (NO) production on BV-2 microglia cells.

Application of NH2 silica gel column chromatography using CH2Cl2-MeOH was found to show a satisfactory resolution for separation of the crude resin glycosides of Pharbitis Semen (the seeds of Pharbitis nil), led to the isolation of seven new acylated glycosidic acid methyl esters, Pharesinosides A-G (1–7), along with four known ones (8–11). These compounds (1–11) were considered to be generated via methyl esterification of the carboxyl group in acylated glycosidic acids. Their structures including stereochemistry were elucidated on the basis of a combination of the spectroscopic and chemical methods. All isolates were evaluated for anti-tumor migration activities with human colon cancer cell line HCT-116, and compound 7 exhibited a potent migration inhibitory activity.

An off-on fluorescent probe (CuCIN), coumarin-naphthol linked by an imine group, for detecting Cu2+ in DMSO-HEPES buffer (1:99, v/v) solution was successfully developed. Mechanism studies suggested that CuCIN firstly formed a complex with Cu2+ in a 1:2 metal-to-ligand ratio, and then 60-fold fluorescence enhancement was observed when Cu2+ promoted the imine hydrolysis to release the fluorescence compound 7-diethylaminocoumarin-3-aldehyde. This probe displayed desired properties such as high selectivity toward other metal ions, low detection limit (12.7 nM) and low cytotoxicity. Bio-imaging study revealed that the new probe CuCIN could be applied for imaging living cells with good cell permeability. We hope to apply this probe in the biomedical research field for the imaging of disease-relevant Cu2+.

Heat shock protein 90 (HSP90) is a chaperone protein that has been shown to regulate cancer progression. As a result, HSP90 has emerged as an attractive target for cancer therapy. Tubocapsenolide A (TA) is an anti-tumor component isolated from Tubocapsicum anomalum. Although the anti-tumor activity of TA was considered to be related to HSP90, the binding site and deep anti-tumor mechanisms still need to be elucidated. In this study, we found that TA is a covalent inhibitor of HSP90, which inhibits HSP90 ATPase activity without blocking ATP binding. Further studies indicated that TA targets the C-terminal Cys521 site, which led to HSP90 partial oligomerization and hindered its anti-aggregation and refolding activity. The damage of the chaperone activity disrupted the interaction between HSP90 and its cochaperone CDC37 as well as its client proteins, thereby inducing cell cycle arrest and apoptosis. Moreover, TA was found to have therapeutic effects on the xenograft tumor model by inducing the degradation of HSP90 client proteins. Together, our results identified HSP90 as the direct target of TA for mediating the anti-tumor activity. TA could serve as a lead compound for developing novel HSP90 C-terminal covalent inhibitors with binding site different from the ATP-binding domain.

Taxodinoid A (1), an unprecedented bis-diterpenic anhydride featuring a 6/6/7/6/6/6 heptacyclic core skeletons, and three novel pyrane-fused abietane-type diterpene dimers (2–4), were obtained from the seeds of Taxodium distichum. Their structures were elucidated by spectroscopic data, X-ray crystallograph and electronic circular dichroism calculation. A biosynthetic pathway and suppressing migration activity against U2 OS cells were also discussed.

To develop novel GLS1 inhibitors as effective therapeutic agents for triple-negative breast cancer (TNBC), 25 derivatives were synthesized from the natural inhibitor withangulatin A (IC50 = 18.2 μM). Bioassay optimization identified a novel and selective GLS1 inhibitor 7 (IC50 = 1.08 μM). In MDA-MB-231 cells, 7 diminished cellular glutamate levels by blocking glutaminolysis pathway, further triggering the generation of reactive oxygen species to induce caspase-dependent apoptosis. Molecular docking indicated that 7 interacted with a new reacting site of allosteric binding pocket by forming various interactions in GLS1. The intraperitoneal administration of 7 at a dose of 50 mg/kg exhibited remarkable therapeutic effects and no apparent toxicity in the MDA-MB-231 xenograft model, indicating its potential as a novel GLS1 inhibitor for treatment of TNBC.

Four new lignans, patulinones A−D (1–4) and three new acetophenone derivatives, patulinones E−G (5–7) were isolated from the leaves of Melicope patulinervia. Their structures were elucidated on the basis of the interpretation of HR-ESIMS, NMR, CD data. All the isolated compounds were evaluated for α-glucosidase inhibitory activity. Of the isolates, compound 4 was found to exhibit the strongest inhibition against α-glucosidase with IC50 value of 6.02 ± 0.46 μM.

Cu(II) complexed amino acid ionic liquid, Cu(II)-[1-butyl-3-methylimidazolium][L-Pro] (Cu(II)-[BMIm][L-Pro]), was successfully adopted as chiral ligand to improve the enantioseparation efficiency in high speed counter-current chromatography (HSCCC). For the enantioseparation of intractable naringenin (NRG) racemic mixtures, Cu(II)-[BMIm][L-Pro] coupled with hydroxypropyl-β-cyclodextrin (HP-β-CD) was successfully applied as dual chiral selectors in HSCCC. The influence of important parameters, including the concentration of the chiral selectors, the pH value, and the temperature were investigated. Under optimal conditions, 4.5mg of (+)-NRG and 4.1mg of (-)-NRG were successfully separated from 10mg NRG racemic mixtures with the purity of 98%. The chiral recognition mechanism of dual chiral selectors was illuminated by the UV-vis and NMR spectra, suggesting that the enantioseparation was upon the difference of the thermodynamic stability of the quaternary complexes of Cu(II), [BMIm][L-Pro], HP-β-CD, and NRG. The results illustrated that the developed HSCCC system, based on the synergistic mechanism of Cu(II)-[BMIm][L-Pro] and HP-β-CD, exhibited better performance on enantioseparation and had great application potential in preparative chiral separation of natural products.

Seven new cadinane-type sesquiterpenoids (1–7) were isolated from the rhizomes of Stahlianthus involucratus. The planar structures of 1–7 were elucidated on the basis of extensive spectroscopic and HRMS data. The absolute configuration of 1 was determined by the single-crystal X-ray diffraction, whereas those of C-9 in 3, C-9 in 4, C-6 in 6, and the 1,2-diol in 7 were deduced via the CD data of the in situ formed [Rh2(OCOCF3)4] complex, and Mo2(OAc)4-induced CD analysis, respectively.

Ten new (1-10) and seventeen known (11-27) neo-clerodane diterpenoids substituted with nicotinoyloxyl were isolated from the plant Scutellaria barbata and their structures were established by extensive spectroscopic analysis. Chemoreversal effects of these neo-clerodane diterpenoids on multidrug resistance were evaluated in breast cancer multidrug-resistant MCF-7/ADR cells that overexpress P-glycoprotein. Four compounds (11, 14, 16, and 18) exhibited better chemoreversal abilities than the classical P-gp inhibitor verapamil and the most potent compound 11 reduced IC50 value of adriamycin in MCF-7/ADR cells from 58.8 μM to 1.3 μM. Mechanistic investigations showed that compound 11 reversed multidrug resistance through suppressing the activity of P-gp and restraining the expression of P-glycoprotein. In the present study, the structure-activity relationships of neo-clerodane diterpenoids were also discussed.

Ten new sucrose esters, physakengoses A-J (1-10), were isolated from the aerial parts of Physalis alkekengi var. franchetii under the guidance of 1H NMR spectroscopy. Their structures were determined by spectroscopic analyses (HRESIMS, 1D and 2D NMR, and ESIMS) and chemical methods. These new compounds were tested for antibacterial activities against Staphylococcus aureus, Bacillus subtilis, Pseudomonas aeruginosa and Escherichia coli. Among them, compounds 2 and 5-8 showed potent inhibitory effects against test strains with MIC values ranging from 3.5 to 14.9μg/mL. These findings may indicate that the P. alkekengi var. franchetii has potential application as an ingredient in pharmaceuticals.

Two new polycyclic polyprenylated acylphloroglucinols, attenuatumione G (1) and attenuatumione H (2), together with five known compounds (3–7) have been isolated from the whole plant of Hypericum attenuatum. Their structures were elucidated by spectroscopic data including HRESIMS, 1D- and 2D-NMR. In biological assays, compound 2 showed moderate cytotoxic activities against Hep-G2 (IC50 9.11 μM) and MCF-7 (IC50 16.24 μM), compound 4 exhibited significant cytotoxic activity with IC50 value of 5.32 μM against Hep-G2, and both compounds 5 and 6 revealed significant nitric oxide production inhibitory activity with IC50 values of 5.52 and 8.46 μM, respectively.

Nine new withaphysalin-type withanolides (1–9), named physaminimins (G-O) were isolated from the whole plants of Physalis minima Linn. The structures of these compounds were elucidated based on spectroscopic methods (1D NMR, HSQC, HMBC, ROESY) and HRESIMS. Physaminimins G, H, and K showed inhibitory activities against lipopolysaccharide (LPS)-induced nitric oxide (NO) production in a macrophage (RAW264.7) cells with IC50 values of 17.41 ± 1.04, 36.33 ± 1.95 and 21.48 ± 1.67 μM, respectively.

Background Therapeutic failure and drug resistance are common and have important implications in the poor prognosis of advanced breast cancer. It is necessary to acquire a natural product to overcome the resistance of cancer and increase the sensitivity of drug-resistant cells to anticancer agents. Purpose To demonstrate whether the compound Vielanin K (VK) has the potential to increase the sensitivity of MCF-7 and MCF-7/MDR cells to anticancer agents. Methods Cell viability and proliferative capacity were determined by MTT, colony formation and EdU assays. Apoptosis and Ca2+ accumulation were evaluated by flow cytometry. Then, proteins were detected by immunoblotting, and gene expression levels were explored by qRT-PCR. Results In MCF-7 and corresponding MDR cells, VK increased the fluorescence intensity of Rho123, but not CFDA. VK treatment did not affect the protein expression of P-gp, MRP1 or BCRP. VK treatment enhanced the DOX-induced apoptotic cascade, while VK combined with DOX increased JNK phosphorylation by activating the IRE1α-TRAF2 signaling pathway. In addition, Ca2+ was released from the endoplasmic reticulum following combination treatment, thereby giving rise to mitochondrial apoptosis. Silencing IRE1α and JNK with small interfering RNA (siRNA) efficiently attenuated combination treatment-induced apoptosis. These effects caused mitochondrial depolarization and reduced viability in MCF-7 and corresponding MCF-7/MDR cells. Conclusion VK combined with DOX increases the apoptosis of MCF-7 and corresponding MCF-7/MDR cells by activating ER stress and mitochondrial apoptosis via IRE1α-TRAF2-JNK signaling.

Hyperpatulones A (1) and B (2), two new polycyclic polyprenylated acylphloroglucinols (PPAPs), were isolated from the leaves of Hypericum patulum. Their unusual 6,7,5-tricyclic system with a peroxide bridge structures were elucidated by comprehensive analysis of their spectroscopic and HRESIMS data. The absolute configuration of hyperpatulone A was determined by comparison between experimental and calculated electron circular dichroism (ECD) spectra. Compounds 1 and 2 exhibited moderate α-glucosidase inhibitory activity with IC50 values of 81.45 ± 1.04 and 12.81 ± 2.13 μM, respectively. Molecular docking studies of 1 and 2 with human maltase-glucoamylase (MGAM) revealing that 2 showed higher shape complementarity with the binding pocket.

Seven new taxane diterpenoids taxumarienes A–G (1–7) were isolated from the leaves of Taxus mairei, along with seven known analogous (8–14). The structures of the new compounds were elucidated based on the analysis of NMR and MS spectroscopy. All isolates were evaluated for their α-glucosidase inhibitory activities. Among them, taxumarienes A (1) and F (6) showed potent effect with IC50 values of 5.9 ± 1.30 μM and 3.7 ± 0.75 μM, respectively.

Water spinach (I. aquatica) is one of the common green leafy vegetables used in Asia. In a continuing chemical investigation on the cytotoxic resin glycosides from water spinach, eight new resin glycosides, Aquaterins XII–XIX (1–8), were isolated, among which 1, 2 and 4 exhibited potent cytotoxic activities. To further characterise the anticancer potential of the resin glycosides from water spinach, Aquaterin II obtained previously was investigated on HepG2 cells. The results showed that Aquaterin II induced G0/G1 arrest regulated by related proteins CDK4/6, cyclin D/E and p21, and caused mitochondria-mediated apoptosis featured by MMP decrease, ROS accumulation, caspase cascade activation and Bax/Bcl-2 alteration. Additionally, the suppression on PI3K/Akt signalling pathway was observed, suggesting that it participated in the Aquaterin II-induced cell growth inhibition. Our results are the first to demonstrate the antiproliferative mechanism of resin glycosides. This investigation also advanced the potential application of water spinach as a functional food.

Seven neo-clerodane diterpenes, teufruintins A-G (1-7), together with eight known compounds (8-15) were isolated from the CHCl3-soluble fraction of the aerial parts of Teucrium fruticans cultivated in China. The chemical structures of the isolated compounds were elucidated using different spectroscopic methods. All of the isolated diterpenes were evaluated for their cytotoxic activities on three human cancer cell lines, and for their ability to inhibit LPS-induced nitric oxide production in RAW 264.7 macrophages. None of the compounds displayed cytotoxic activities on the cancer cell lines, and only 15 showed weak NO inhibitory activity.

Two new 18,19-secoursane triterpenes and three triterpene glucosides, together with twelve known compounds, were isolated from the aerial parts of Rubus lambertianus Ser. The structures of the new compounds were determined by spectroscopic analyses (HR-ESI-MS, 1D/2D NMR). The protective effects of compounds 1–17 on high glucose-induced oxidative stress injury in human umbilical vein endothelial cells (HUVECs) were evaluated using cell viability assay. Compounds 1, 2, 7, 10, 12, 15 and 17 significantly attenuated high glucose-induced oxidative damage on HUVECs at the concentration of 3.10 μM.

A pair of 2,3-seco cafestol-type diterpenoid epimers, frutilactones A (1) and B (2) were isolated from twigs of Tricalysia fruticosa. Their structures were elucidated by comprehensively employing spectroscopic methods (MS and 1D/2D NMR) and ECD. Their inhibitory effects on nitric oxide production in lipopolysaccharide-activated macrophages were evaluated, and a possible biogenetic pathway was also proposed.

The use of a dinuclear Cu2(II)-β-cyclodextrin complex (Cu2-β-CyD) as a chiral selector (CS) was successfully applied to chiral high speed counter-current chromatography (HSCCC) for the enantioseparation of aromatic α-hydroxycarboxylic acids. α-cyclohexylmandelic acid (CHMA) as a case was studied to optimize the separation temperature, the pH value of the aqueous phase, and the concentration of the chiral selector, which were comparatively studied between Cu2-β-CyD and native β-cyclodextrin (β-CyD) as CSs. The thermodynamic parameters of the formation of the complex between the enantiomers and the CSs were obtained, and the complex formation constants between the enantiomers and Cu2-β-CyD or β-CyD were determined using induced circular dichroism technology. The results indicate that Cu2-β-CyD can remarkably improve the enantioseparation compared with β-CyD, mainly due to the ion-pairing interactions between the deprotonation of carboxyl group of CHMA and the Cu(II) ion of Cu2-β-CyD. Five of other aromatic α-hydroxycarboxylic acids were also enantioseparated successfully by HSCCC using Cu2-β-CyD as a CS.

(±)-Calliviminones A (1) and B (2), two Diels–Alder adducts of polymethylated phloroglucinol and myrcene with unprecedented spiro-[5.5] undecene skeleton, were isolated from the fruits of Callistemon viminalis. Structural elucidation was accomplished by NMR spectra studies, and the biomimetic synthesis of compounds 1 and 2 confirmed the pivotal role of Diels–Alder reaction in the plausible biosynthetic pathway. Compounds 1 and 2 were also the first example of Carbon Diels–Alder adducts between phloroglucinol and terpene. Bioactivity scan indicated that 1 and 2 showed moderate inhibition on NO production on lipopolysaccharide-induced RAW264.7 macrophages.

Ten new calyxin natural products, named calyxin N (1), ent-calyxin N (2), calyxin O (3), ent-calyxin O (4), calyxin P (5), 9″-epicalyxin P (6), calyxin Q (7), calyxin R (8), calyxin S (9) and 5-epicalyxin S (10), were isolated from the seeds of Alpinia katsumadai. Their planar structures were elucidated by a combination of various spectroscopic methods, primarily NMR and MS, while the absolute configurations of the isolated calyxins were comprehensively studied by the modified Mosher's method, electronic circular dichroism (ECD) and theoretical calculations. This is the first systematically study on the absolute configurations of calyxins. Most of the isolated compounds showed moderate to strong antiproliferative activities against NCI-H460, HeLa, SMMC-7721 and HCT-116 cancer cell lines.

Six new withanolides (1-6), including two uncommon 1,10-seco withanolides (1 and 2), together with five known withanolides (7-11), were isolated from the whole plants of Physalis minima Linn.. The structures of new compounds were elucidated through spectroscopic methods, including (1)H, (13)C NMR, 2D-NMR, HRESIMS and circular dichroism (CD). Inhibitory effects of the isolates on nitric oxide (NO) production in lipopolysaccaride-activated RAW264.7 macrophages were evaluated. Compounds 2 and 5 showed strong inhibitory activities with IC50 values of 8.04 and 10.01 μM, respectively. Compounds 1, 9 and 10 exhibited moderate inhibitory activities with IC50 values from 25.54 to 43.58 μM.

Two new resin glycosides, cuses 3 (1) and 4 (2), were initially obtained from the seeds of Cuscuta chinensis. Both of them contained a reducing glucose unit, and thus existed as a pair of isomers. In order to solve the existing problem of tautomerism, the remanent resin glycoside fraction was converted into aminoalditol derivatives with p-anisidine, and then another eight new resin glycosides cuses 5–12 (3–10) were further isolated. Cuses 7–12 (6–10) were considered to be generated via glycosidation of two acylated oligosaccharides, and thus characterized as ether-type resin glycoside dimers. Their structures including absolute stereochemistry were elucidated by a combination of spectroscopic and chemical methods. Compounds 3–10 exhibited cytotoxic activity toward MCF-7, SMMC-7721, and MG-63 cell lines with IC50 values ranging from 8.72 to 59.35 μg/mL.

Panax ginseng C.A. Meyer has been known as a valuable traditional Chinese medicines for thousands years of history. Ginsenosides, the main active constituents, exhibit prominent immunoregulation effect. The present study first describes a holistic method based on chemical characteristic and lymphocyte proliferative capacity to evaluate systematically the quality of P. ginseng in thirty samples from different seasons during 2-6 years. The HPLC fingerprints were evaluated using principle component analysis (PCA) and hierarchical clustering analysis (HCA). The spectrum-efficacy model between HPLC fingerprints and T-lymphocyte proliferative activities was investigated by principal component regression (PCR) and partial least squares (PLS). The results indicated that the growth of the ginsenosides could be grouped into three periods and from August of the fifth year, P. ginseng appeared significant lymphocyte proliferative capacity. Close correlation existed between the spectrum-efficacy relationship and ginsenosides Rb1, Ro, Rc, Rb2 and Re were the main contributive components to the lymphocyte proliferative capacity. This comprehensive strategy, providing reliable and adequate scientific evidence, could be applied to other TCMs to ameliorate their quality control.

Melicolones C–K (1–9), nine acetophenone derivatives with three unprecedented architectures, were characterized from the leaves of Melicope ptelefolia. Among them, melicolone C (1) was a novel acetophenone analogue possessing a highly rearranged spiro skeleton. Melicolones D-F (2–4) were three novel acetophenone congeners bearing unusual octalactone ring. Melicolones G-K (5–9) were five new prenylated acetophenone derivatives featuring a 9-oxatricyclo [3.2.1.13, 8]nonane core. Their structures were established by extensive spectroscopic analysis and ECD spectra. These different structures were presumably derived from the same precursor via three major biosynthetic pathways. Compounds 5–9 exerted moderate effects to reverse multidrug-resistance in MCF-7/ADR cells with reversal fold values ranging from 6.2 to 13.3.

Aim To evaluate the radiological and histopathological features of hepatic inflammatory myofibroblastic tumours (IMTs), and improve the understanding of this tumour. Materials and methods A retrospective analysis of radiological and histopathological features of 10 cases of IMT was carried out from May 2003 to September 2011 at the Second Xiangya Hospital of Central South University. Results Ten cases (five male and five female patients; age range 4–68 years) were enrolled. Computed tomography (CT) and magnetic resonance imaging (MRI) revealed that the lesions were regular, hypodense or hypointense (T1WI) masses with well-defined borders ( n = 8) or ill-defined borders ( n = 2). The maximum diameter ranged from 3.1–13.4 cm (mean = 6 cm). The masses showed homogeneous ( n = 8) or inhomogeneous ( n = 2) density. Contrast-enhanced CT and MRI showed the lesions were mildly, irregularly enhanced ( n = 7) or not enhanced ( n = 2) in the arterial phase and markedly enhanced in the portal venous phase or delayed phase. Hepatic arteriography revealed that the lesions were hypovascular and had a well-defined border. One patient had lung metastasis with obvious arterial phase enhancement. None of the patients had a history of hepatitis, cirrhosis, or enlarged lymph nodes. Pathology showed that the gross appearance of the tumours was smooth. The tumour cells comprised spindle-shaped fibroblast and myofibroblast cells with abundant inflammatory cells. Immunohistochemistry showed that most were positive for vimentin, smooth muscle actin (SMA), and CD68, but negative for CD34, anaplastic lymphoma kinase (ALK), S-100, and CD117. Conclusion Radiological features of IMT have some characteristics of an intermediate-grade malignant tumour. However, imaging alone cannot be used to diagnose IMT. Therefore, histopathological examination is necessary for confirmation.

• Two triterpenoids, fourteen lignans and three phenolics were obtained from Boschniakia himalaica .

Two new diarylheptanoids with a tetrahydropyran ring, kravanhol A (1) and kravanhol B (2), along with one known diarylheptanoid renealtin A (3) were isolated from the fruits of Amomum kravanh. The structures of compounds 1 and 2 were established by analysis of spectroscopic data and their absolute configurations were determined by Mosher's method and CD experiments. Compound 2 showed inhibitory effect on nitric oxide production in lipopolysaccharide-activated RAW264.7 macrophages with an IC50 value of 38.9 ± 1.8 μM.

Assays based on high-performance liquid chromatography (HPLC) and liquid chromatography tandem mass spectrometry (LC–MSn) have been developed and validated for the determination and metabolite identification of the bidesmosidic triterpenoid saponin, BTS-1 (3-O-β- d -galactopyranosyl-(1→2)-[β- d -xylopyranosyl-(1→3)]-β- d -glucuronopyranosyl gypsogenin 28-O-α- l -arabinopyranosyl-(1→3)-β- d -xylopyranosyl-(1→4)-α- l -rhamnopyranosyl-(1→2)-β- d -fucopyranoside), in rat plasma. The assay was successfully applied to a pharmacokinetic study in rats given a single oral dose of BTS-1 (400 mg/kg). The results indicated that the compound was rapidly absorbed (Tmax=1.28±0.29 h, Cmax=37.4±5.6 µg/mL) and slowly eliminated (t1/2=13.2±6.6 h). In addition, secondary glycosides and aglycones of BTS-1 were detected and identified. Since these metabolites are known to be active α-glucosidase inhibitors, they probably play an important role in mediating the pharmacological effects of the saponin.

Two new β-carboline-type alkaloids, dichotomine K (1) and dichotomine L (2), were isolated from the roots of Chinese medicinal plant Stellaria dichotoma L. var. lanceolata Bge. Structures of 1 and 2 were determined on the basis of chemical and spectroscopic means.

Phytochemical investigation of the bulbs of Lilium brownii var. viridulum led to the isolation of seven new steroidal saponins (1-7), along with eight known analogues (8-15). The new steroidal saponins were identified as 27-O-[(3S)-3-O-β-D-glucopyranosyl 3-methylglutaroyl]isonarthogenin 3-O-[α-L-rhamnopyranosyl-(1→2)]-β-D-glucopyranoside (1), (24S,25S)-3β,17α,24-trihydroxy-5α-spirostan-6-one 3-O-[α-L-rhamnopyranosyl-(1→2)]-β-D-glucopyranoside (2), tenuifoliol 3-O-[β-D-glucopyranosyl-(1→4)]-β-D-glucopyranoside (3), 26-O-β-D-glucopyranosylnuatigenin (4), 26-O-β-D-glucopyranosylnuatigenin 3-O-β-D-glucopyranoside (5), 26-O-β-D-glucopyranosylnuatigenin 3-O-{α-L-rhamnopyranosyl-(1→2)-[β-D-glucopyranosyl-(1→6)]}-β-D-glucopyranoside (6), 26-O-[β-D-glucopyranosyl-(1→2)]-β-D-glucopyranosylnuatigenin 3-O-[α-L-rhamnopyranosyl-(1→2)]-β-D-glucopyranoside (7), using a combination of spectroscopic evidence and chemical methods. The carbohydrate chain of a sugar linked to C-3 of the HMG group (3-hydroxy-3-methylglutarate) in compound 1 is rarely found in nature. Compound 2 possesses a new (24S,25S)-3β,17α,24-trihydroxy-5α-spirostan-6-one aglycon moiety. The disaccharide chain linked to C-26 hydroxy group of the furospirostanol in compound 7 has not been observed from natural sources.

A novel on-line method using high performance liquid chromatography (HPLC) with a diode array detector (DAD) and a chemiluminescence (CL) detector was successfully developed for simultaneously analyzing the antioxidant efficacies of 15 components in Guan-Xin-Ning (GXN) injection, a traditional Chinese medicinal preparation consisting of Radix Salvia miltiorrhiza and Rhizoma Ligusticum chuanxiong for the treatment of coronary heart disease and angina pectoris. The analytical conditions were optimized, validated and applied to determine H 2 O 2 inhibition rates of the 15 phenolic acids in GXN injections from 14 different production batches. The results showed that antioxidant diversities were found in the 15 compounds and that danshensu, protocatechuic aldehyde and rosmarinic acid were largely responsible for the antioxidant activity of GXN injection (25.82%-51.19%). In addition, 14 batches of GXN injections were divided into five groups corresponding with five different factories by principal component analysis (PCA) assay. Therefore, the proposed HPLC-DAD-CL method was simple, rapid and low-cost for screening antioxidant constituents in a complex system, which can be used to reveal the antioxidant mechanism of GXN injection and is applicable for its quality control in terms of activity, which was different from the routine chemical assessment.

Two new compounds, a neolignan (1), and a benzofuran derivative (2), along with 6 known compounds (3-8), were isolated from the aerial parts of Leontopodium leontopodioides. The structures of the new compounds were elucidated as (7R,8S)-3,5'-dimethoxy-4',7-epoxy-8,3'-neolignane-5,9,9'-triol (1) and (2R)-12-hydoxy-4-methoxy-tremeton (2) on the basis of their spectroscopic data, respectively. All of the isolates were evaluated for their effects on the inhibition of nitric oxide (NO) production in lipopolysaccharide-activated RAW264.7 macrophages and compounds 1 and 8 exhibited inhibitory activity with IC(50) values of 35.80 and 24.41 μM, respectively.

Preparative separation of seven phenylpropenoid glycerides from the bulbs of Lilium lancifolium (lily), was conducted by high-speed counter-current chromatography (HSCCC) with a two-phase solvent system composed of n -hexane–ethyl acetate–methanol–0.05% aqueous trifluoroacetic acid (TFA) (3:5:3:5, v/v/v/v). These phenylpropenoid glycerides were identified as 1- O -feruloylglycerol (1), 1- O - p -coumaroylglycerol (2), 1- O -caffeoyl-3- O - p -coumaroylglycerol (3), 1,2- O -diferuloylglycerol (4), 1,3- O -diferuloylglycerol (5), 1- O -feruloyl-3- O - p -coumaroylglycerol (6), and 1,3- O -di- p -coumaroylglycerol (7), respectively, by ESI-MS, 1 H NMR and 13 C NMR. Their antioxidant activities were evaluated by DPPH radical scavenging activity, ABTS radical cation scavenging activity, and ferric-reducing antioxidant power (FRAP). The trend in antioxidant capacity was similar in all the three assays, with 3 > 1, 4, 5, 6 > 2, 7. This is the first report on simultaneous separation of seven antioxidantive phenylpropenoid glycerides from L. lancifolium by HSCCC.