Alkaloids from the stems of Fissistigma maclurei Merr. inhibit the proliferation of synoviocytes.

Two new aporphine-derived alkaloids, aporaloids C and D (1 and 2), along with eight known biogenetically related alkaloids (3-10) were isolated from the stems of Fissistigma maclurei Merr. Their structures were elucidated by detailed analysis of NMR, HRESIMS, MS, IR, UV and Optical rotations data. Compounds 1 and 2 represent a rare example of N-methylol aporphine-derived alkaloids from natural sources. The inhibitory effect of all compounds on the proliferation of primary synovial cells was evaluated. Compound 3 showed potent inhibitory effect on the proliferation of synoviocytes with an IC50 value of 4.8 µM. Compounds 1, 2, 6-9 and 10 exhibited moderate inhibitory activity on synoviocytes, with IC50 values of 36.8, 37.1, 31.2 µM, 32.5, 36.3, 36.8 and 18.2 µM, respectively.

Two new anthraquinone derivatives from Saprosma crassipes H. S. Lo.

Two new anthraquinone derivatives sapranquinones A and B (1 and 2) together with two known biogenetically related anthraquinone derivatives (3 and 4) were isolated from the stems of Saprosma crassipes H. S. Lo. The structures of these compounds were elucidated using comprehensive spectroscopic methods. Compounds 1-4 were evaluated for their antibacterial activities and compounds 1 and 3 had a broad spectrum antibacterial activity against Staphylococcus albus, Escherichia coli, Bacillus cereus, Micrococcus tetragenus, and Micrococcus luteus with MIC values ranging from 1.25 to 5 µg/mL.

Flavonoidal alkaloids from the flowers of Chromolaena odorata (L.) R.M.King & H.Rob.

A pair of epimers of flavonoid alkaloids, with a pyrrolidone moiety, 2S,5''R-eupodoratin A (1), 2S,5''S-eupodoratin A (2), together with two known analogues, drahebephin A (3), drahebephin B (4), were isolated from the flowers of Chromolaena odorata (L.) R.M.King & H.Rob. Their structures were elucidated on the basis of HR-ESI-MS, 1D/2D NMR spectral analyses. The absolute configuration of compounds (1) and (2) was determined by its experimental and calculated electronic circular dichroism (ECD) spectra. All compounds were isolated from the Asteraceae family for the first time. The ABTS·+ scavenging activity of compound (4) reached 93.56% at a concentration of 0.5 mM, while the scavenging capacity of positive control Trolox was 55.94%. In addition, all compounds show moderate antimicrobial activity against Escherichia coli (ATCC, 337304), Staphylococcus aureus (ATCC, 337371) and Candida albicans (ATCC, 186382) with a MIC value of more than 50 µg/mL.

Isolated compounds from Dracaena angustifolia Roxb and acarbose synergistically/additively inhibit α-glucosidase and α-amylase: an in vitro study.

BACKGROUND: As a traditional herbal medicine, Dracaena angustifolia Roxb has been used as an anti-inflammatory agent by the Li people in Hainan, China. In preliminary phytochemical studies conducted in our lab, its fractions were found to inhibit α-glucosidase in vitro, indicating a potential for alleviating glucose dysregulation. METHODS: Through in vitro enzymatic assays, the abilities of the separated components to affect α-glucosidase and α-amylase were evaluated. By establishing concentration gradients and generating Lineweaver-Burk plots, the corresponding inhibition modes together with kinetic parameters were assessed. Following the evaluation of the outcomes of their combination with acarbose, computational docking and molecular dynamic simulations were carried out to analyse the interaction mechanisms and perform virtual screening against human enzymes. RESULTS: Compared with acarbose, 7 compounds, including flavonoid derivatives, amides and aromatic derivatives, with higher α-glucosidase inhibitory efficiencies were confirmed. It was found that those competitive/mixed candidates and acarbose interacted synergistically or additively on α-glucosidase. Moreover, 3 of them were able to inhibit α-amylase in mixed mode, and additive effects were observed in combination with acarbose. Through in silico docking, it was found that the active site residues as well as adjacent residues were involved in α-glucosidase and α-amylase binding, which were mainly achieved through hydrogen bonding. Among those dual-function flavonoids, Compound 9 was predicted to be a considerable inhibitor of human enzymes, as the formation of ligand-enzyme complexes was mediated by the residues responsible for substrate recognition and catalysis, the stabilities of which were reiterated by molecular dynamics simulations. CONCLUSION: Despite their mild effects on α-amylase, considerable α-glucosidase inhibitory efficiencies and potential synergy with acarbose were exhibited by these natural candidates. Furthermore, a stable ligand, human α-glucosidase, was predicted by the performed simulations, which provided useful information for the application of Dracaena angustifolia Roxb in diabetes treatment.

Fissistiganoids A and B: two new flavonoids from the Fissistigma tungfangense.

Two new flavanoids fissistiganoids A and B (1 and 2), together with two known pterocarpans derivatives (3 and 4), were isolated from the stems of Fissistigma tungfangense. The structures of these compounds were elucidated using comprehensive spectroscopic methods. The absolute configurations of fissistiganoids A and B (1 and 2) were determined by comparing their ECD spectra with quantum-mechanics ECD calculations. The inhibitory activities of all compounds against three cancer cell lines HeLa, MCF-7 and A549 were evaluated. Compounds 1-4 showed moderate inhibitory effects on HeLa, MCF-7 and A549 cells with IC50 values ranging from 12.5 to 42.3 µM.

Two new bioactive secondary metabolites from the endophytic fungus Talaromyces assiutensis JTY2.

One new lactone derivative helicascolide F (1), one new pyrrolidine derivative talaromydine (2), along with six known compounds (3-8) were isolated from the fungus Talaromyces assiutensis JTY2. The structure of the new compounds 1 and 2 was determined by 1D and 2D NMR as well as by HRESIMS. The inhibitory activity of all compounds against six phytopathogenic fungi and three cancer cell lines was evaluated.

Five New Triene Derivatives from the Fungus Penicillium herquei JX4.

Five undescribed triene derivatives, pinophols B-F (2-6), together with one known compound, pinophol A (1), were obtained from the mangrove endophytic fungus Penicillium herquei JX4. The structures of compounds 1-6 were elucidated using IR, HR-ESI-MS, and NMR methods. The absolute configurations of compounds 1-6 were confirmed by comparing their experimental or calculated ECD spectra. Pinophols C and D (3 and 4) showed inhibitory activities against LPS-induced NO production.

Synthesis and assessment of CTAB and NPE modified organo-montmorillonite for the fabrication of organo-montmorillonite/alginate based hydrophobic pharmaceutical controlled-release formulation.

The research goal of the present study was to develop a carrier for loading and controlled -release of the hydrophobic drug with the combined use of organo-montmorillonite (OMMT) and alginate. The OMMT was synthesized through the intercalation modification of sodium montmorillonite (Na-MMT) with cationic cetyltrimethylammonium bromide (CTAB), nonionic nonylphenol polyoxyethylene ether (NPE) and the mixture of them via simple and convenient wet ball-milling method. Furthermore, the organo-montmorillonite/alginate (OMMT/Alg) composite hydrogel beads with slow and controlled release properties were constructed by using alginate as a coating material under the exogenous cross-linking of calcium ions. The physical and chemical properties of OMMT were comparatively evaluated by scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FT-IR), X-ray diffraction (XRD), transmission electron microscopy (TEM), dynamic light scattering (DLS), thermogravimetric analyzer (TGA), BET-specific surface area measurements, and drug adsorption experiments. Experimental results showed that the presence of CTAB was able to facilitate the intercalation of CTAB/NPE into Na-MMT through the cation exchange reaction. And the cationic CTAB and nonionic NPE were adsorbed or intercalated into the MMT lamellar structure through the wet ball-milling process, which could change the hydrophilic nature of Na-MMT and improve its affinity to the hydrophobic drug molecules. In addition, the OMMT/Alg composite hydrogel beads displayed superior sustained-release properties than Na-MMT/Alg, mainly ascribed to the good affinity of OMMT to hydrophobic drug that retarded the drug diffusion. In particular, CTA/NPE-MMT/Alg with the highest loading capacity (LC) and encapsulation efficiency (EE) revealed the optimal controlled performance for the release of hydrophobic ibuprofen. The release followed the Korsmeyer-Peppas model suggested non-Fickian diffusion release mechanism. Based on the high drug loading capacity and excellent controlled drug release properties, the CTA/NPE-MMT/Alg incorporating hydrophobic drugs into hydrophilic matrices could be a highly promising material for use in hydrophobic drug delivery.

In vitro and in vivo mechanism of hepatocellular carcinoma inhibition by ß-TCP nanoparticles.

Background: Studies have showed that nanoparticles have a certain anti-cancer activity and can inhibit many kinds of cancer cells. ß-tricalcium phosphate nanoparticles (nano-ß-TCP) displays better biodegradation, but the application and mechanism of nano-ß-TCP in anti-cancer activity are still not clear. Purpose: The objective of this study was to synthesize nano-ß-TCP and investigate its inhibitory properties and mechanism on hepatocellular carcinoma (HepG2) cells in vitro and in vivo. Methods: Nano-ß-TCP was synthesized using ethanol-water system and characterized. The effects of nano-ß-TCP on cell viability, cell uptake, intracellular oxidative stress (ROS), cell cycle and apoptosis were also investigated with HepG2 cells and human hepatocyte cells (L-02). Intratumoral injection of nano-ß-TCP was performed on the xenograft liver cancer model to explore the inhibitory effect and mechanism of nano-ß-TCP on liver tumors. Results: In vitro results revealed that nano-ß-TCP caused reduced cell viability of HepG2 cells in a time-and dose-dependent manner. Nano-ß-TCP was internalized through endocytosis and degraded in cells, resulting in obvious increase of the intracellular Ca2+ and PO4 3- ions. Nano-ß-TCP induced cancer cells to produce ROS and induced apoptosis of tumor cells by an apoptotic signaling pathways both in extrinsic and intrinsic pathway. In addition, nano-ß-TCP blocked cell cycle of HepG2 cells in G0/G1 phase and disturbed expression of some related cyclins. In vivo results showed that 40 mg/kg of nano-ß-TCP had no significant toxic side effects, but could effectively suppress hepatocellular carcinoma growth. Conclusion: These findings revealed the anticancer effect of nano-ß-TCP and also clarified the mechanism of its inhibitory effect on hepatocellular carcinoma.

Citrate reduced oxidative damage in stem cells by regulating cellular redox signaling pathways and represent a potential treatment for oxidative stress-induced diseases.

Chemical substances containing citrate such as calcium citrate, citrate esters and citric acid exhibit anti-oxidant and anti-inflammatory properties in different cells and tissues. However, data on the anti-oxidant and anti-inflammatory properties and mechanisms of action of citrate are insufficient. In this study, we systematically evaluated the anti-oxidant capacity of citrate using chemical, cellular and animal assays. Citrate showed a stable molecular structure and did not directly react with oxides. Citrate exerted protective and anti-apoptotic effects on BMSCs and also showed significant inhibitory effects on the oxidative stress and inflammatory reactions in the rat air pouch model. By using proteomics, we found that PPARγ contributed to the upregulation of various free radical scavenging proteins and the downregulation of diverse components of the inflammatory responses. Citrate-regulated global PPARγ expression was evidenced by the significant increase expression of PPARγ in PC12 cell line. Our results provide novel insights into the role of citrate in regulating cellular redox signaling and the function of PPARγ signaling in this process and also provide basic molecular cell biology information to improve the applications of biomaterials or stem cells as treatments for oxidative stress-induced degenerative diseases and inflammatory diseases.

Painful Terminal Neuroma Prevention by Capping PRGD/PDLLA Conduit in Rat Sciatic Nerves.

Neuroma formation after amputation as a long-term deficiency leads to spontaneous neuropathic pain that reduces quality of life of patients. To prevent neuroma formation, capping techniques are implemented as effective treatments. However, an ideal, biocompatible material covering the nerves is an unmet clinical need. In this study, biocompatible characteristics presented by the poly(D,L-lactic acid)/arginylglycylaspartic acid (RGD peptide) modification of poly{(lactic acid)-co- [(glycolic acid)-alt-(L-lysine)]} (PRGD/PDLLA) are evaluated as a nerve conduit. After being capped on the rat sciatic nerve stump in vivo, rodent behaviors and tissue structures are compared via autotomy scoring and histological analyses. The PRGD/PDLLA capped group gains lower autotomy score and improves the recovery, where inflammatory infiltrations and excessive collagen deposition are defeated. Transmission electron microscopy images of the regeneration of myelin sheath in both groups show that abnormal myelination is only present in the uncapped rats. Changes in related genes (MPZ, MBP, MAG, and Krox20) are monitored quantitative real-time polymerase chain reaction (qRT-PCR) for mechanism investigation. The PRGD/PDLLA capping conduits not only act as physical barriers to inhibit the invasion of inflammatory infiltration in the scar tissue but also provide a suitable microenvironment for promoting nerve repairing and avoiding neuroma formation during nerve recovery.

The effect of different hydroxyapatite microparticles on the osteogenic differentiation of MC3T3-E1 preosteoblasts.

Hydroxyapatite (HA) has been extensively explored for biomedical applications due to its advantages of facile synthesis and surface functions. Many studies have suggested that HA microparticles can induce osteogenic differentiation. However, how the size and shape of HA microparticles affected the differentiation response has not been elucidated. In this work, HA microparticles (HA whiskers and HA microspheres) with different sizes of 40, 50 and 80 µm were synthesized and their effects on the osteogenic differentiation of MC3T3-E1 preosteoblasts were investigated. All the HA microparticles showed cytocompatibility and did not show their influence on the proliferation at the studied concentrations. The osteogenic differentiation of MC3T3-E1 preosteoblasts was dependent on the size and shape of HA. Sphere-50 and sphere-80 significantly increased the alkaline phosphatase (ALP) activity, calcium deposition and expression of Runx2 and collagen I, while whisker-40, whisker-50 and whisker-80 reduced the ALP activity and calcium deposition. The results indicated that the size and shape of HA microparticles have an influence on the osteogenic differentiation of MC3T3-E1 preosteoblasts.

Degradation characteristics, cell viability and host tissue responses of PDLLA-based scaffold with PRGD and ß-TCP nanoparticles incorporation.

This study is aimed to evaluate the degradation characteristics, cell viability and host tissue responses of PDLLA/PRGD/ß-TCP (PRT) composite nerve scaffold, which was fabricated by poly(d, l-lactic acid) (PDLLA), RGD peptide(Gly-Arg-Gly-Asp-Tyr, GRGDY, abbreviated as RGD) modified poly-{(lactic acid)-co-[(glycolic acid)-alt-(l-lysine)]}(PRGD) and ß-tricalcium phosphate (ß-TCP). The scaffolds' in vitro degradation behaviors were investigated in detail by analysing changes in weight loss, pH and morphology. Then, the 3-(4,5-dimethyl-2-thiazolyl) -2,5-diphenyl-2 -H-tetrazolium bromide (MTT) assay and cell live/dead assay were carried out to assess their cell viability. Moreover, in vivo degradation patterns and host inflammation responses were monitored by subcutaneous implantation of PRT scaffold in rats. Our data showed that, among the tested scaffolds, the PRT scaffold had the best buffering capacity (pH = 6.1-6.3) and fastest degradation rate (12.4%, 8 weeks) during in vitro study, which was contributed by the incorporation of ß-TCP nanoparticles. After in vitro and in vivo degradation, the high porosity structure of PRT could be observed using scanning electron microscopy. Meanwhile, the PRT scaffold could significantly promote cell survival. In the PRT scaffold implantation region, less inflammatory cells (especially for neutrophil and lymphocyte) could be detected. These results indicated that the PRT composite scaffold had a good biodegradable property; it could improve cells survival and reduced the adverse host tissue inflammation responses.