The inhibition of resveratrol to human skin squamous cell carcinoma A431 xenografts in nude mice

Squamous cell carcinoma (SCC) is one of the commonest dermatological malignancies. Resveratrol (Res) is one type of polyphenolic compound which was first identified from the roots of Veratrum grandinorum in 1940. The previous studies found that Res can promote apoptosis of a variety of tumor cell, especially SCC cells. However it is rare to study the inhibition mechanism of Res in the animal model. In this study, through the establishment of human cutaneous SCC A431 xenografts in nude mice, we observed Res inhibition effect and investigated the inhibition mechanism by checking the expression of apoptosis-related factors, p53, ERK and survivin. The results showed that the xenograft volume and weight of Res groups were less than those of the control groups (P < 0.05), but the net body mass of nude mice of Res groups was not significantly different from the control groups (P > 0.05). The apoptotic index of Res groups were significantly higher than the control groups (P < 0.05). The protein and mRNA expression of p53 and ERK were statistically positively correlated (P < 0.05) and significantly increased in Res high- and medium-dose groups compared with the control groups (P < 0.05). Moreover, the protein and mRNA expression of SVV were negatively correlated with p53 (P < 0.05) and lower than the control groups (P < 0.05). The results demonstrate Res inhibitory effect and indicate that the inhibition mechanism of Res is to upgrade the protein and mRNA expression of p53 and to downgrade the protein and mRNA expression of SVV, thus inducing the apoptosis of tumor cells.     

Hormone-sensitive lipase is involved in the action of hydroxysafflor yellow A (HYSA) inhibiting adipogenesis of 3T3-L1cells

BackgroundSafflor yellow A (SY) has been demonstrated to be beneficial to cardiovascular system. Our previous study showed that hydroxysafflor yellow A (HSYA), a main component of SY, could increase peroxisome proliferator-activated receptor γ mRNA expression. In this study, we investigate the effect of HSYA on the proliferation and adipogenesis of mouse 3T3-L1 preadipocytes.MethodsThe proliferation and adipogenesis of 3T3-L1 cells treated with HSYA was studied by 3-(4,5-dimethylthiazol-2-yl) 2,5-diphenyl tetrazolium bromide (MTT) spectrophotometry, Oil Red O staining and intracellular triglyceride assay methods. HSL mRNA expression and promoter activity were studied by real-time quantitative RT-PCR, transient transfection and dual luciferase reporter gene methods.ResultsHSYA (0.1 mg/L) significantly inhibited the proliferation of 3T3-L1 cells when compared with control cells in 8 h. This effect was further enhanced with the extension time (24 to 96 h) and an increase of concentration of HSYA (1–10 mg/L). The maximal inhibitory action was observed at 0.1 mg/L HSYA in 72 h (86 ± 11.8% vs. 100 ± 4.1%, p < 0.01). HSYA notably reduced the amount of intracellular lipid and triglyceride content in adipocytes to 85% (1 mg/L) and 75% (100 mg/L) on Day 4 following the differentiation, respectively, while increased HSL mRNA expression and promoter activities to 2.7 fold and 1.55 fold, respectively (p < 0.01), in differentiated 3T3-L1 adipocytes.ConclusionsHSYA inhibits the proliferation and adipogenesis of 3T3-L1 preadipocytes. The inhibitory action of HYSA on adipogenesis may be due to the promotion of lipolytic-specific enzyme HSL expression by increasing HSL promoter activity.     

Effects of rhynchophylline on GluN1 and GluN2B expressions in primary cultured hippocampal neurons

N-methyl-d-aspartate (NMDA) receptor subunits GluN1 and GluN2B in hippocampal neurons play key roles in anxiety. Our previous studies show that rhynchophylline, an active component of the Uncaria species, down-regulates GluN2B expression in the hippocampal CA1 area of amphetamine-induced rat. The effects of rhynchophylline on expressions of GluN1 and GluN2B in primary hippocampal neurons in neonatal rats in vitro were investigated. Neonatal hippocampal neurons were cultured with neurobasal-A medium. After incubation for 6 h or 48 h with rhynchophylline (non-competitive NMDAR antagonist) and MK-801 (non-competitive NMDAR antagonist with anxiolytic effect, as the control drug) from day 6, neuron toxicity, mRNA and protein expressions of GluN1 and GluN2B were analyzed. GluN1 is mainly distributed on neuronal axons and dendritic trunks, cytoplasm and cell membrane near axons and dendrites. GluN2B is mainly distributed on the membrane, dendrites, and axon membranes. GluN1 and GluN2B are codistributed on dendritic trunks and dendritic spines. After 48 h incubation, a lower concentration of rhynchophylline (lower than 400 μmol/L) and MK-801 (lower than 200 μmol/L) have no toxicity on neonatal hippocampal neurons. Rhynchophylline up-regulated GluN1 mRNA expression at 6 h and mRNA and protein expressions at 48 h, but down-regulated GluN2B mRNA and protein expressions at 48 h. However, GluN1 and GluN2B mRNA expressions were down-regulated at 6 h, and mRNA and protein expressions were both up-regulated by MK-801 at 48 h. These findings show that rhynchophylline reciprocally regulates GluN1 and GluN2B expressions in hippocampal neurons, indicating a potential anxiolytic property for rhynchophylline.     

Calibrating and using an EM31 electromagnetic induction meter to estimate and map soil salinity in the jarrah and karri forests of south-western Australia

Knowledge of the spatial distribution of soil salinity in areas of jarrah and karri forest where groundwater levels may be close to the surface is important when planning land use activities that may reduce leaf area and thus increase the risk of a rise in stream salinity. Hence this study aimed, firstly, to test whether a Geonics EM31 induction meter could be used to accurately estimate soil salinity in jarrah and karri forest soils. Secondly, to determine an appropriate method of surveying the stream zone with an EM31 to adequately represent the spatial variation in soil salinity.A comparison of EM31 meter estimates of soil apparent electrical conductivity (ECa) with average soil salinity profiles (EC_1:5) to 6 m depth across a broad range of sites resulted in good correlations in spring (R² = 0.66, n = 164, p < 0.001) and autumn (R² = 0.64, n = 156, p < 0.001). ECa measured in spring, when soil moisture was at its highest, was 25% higher than in autumn, when soil moisture was at its lowest.EM31 surveys along second-order stream zones showed that the highest ECa levels generally occurred within 10 m of the stream centre. However, ECa was highly variable along the length of the streams and the highest levels did not necessarily occur at the lowest part of the catchment. A suitable method for conducting surveys of stream zones is to carry an EM31, while walking along a traverse, taking a reading approximately every 2 m.     

Genotype × environment interactions in selected loblolly and slash pine plantations in the Southeastern United States

Few studies have quantified the combined effects of silvicultural treatments and genetic improvement on unit area production of full-sib family blocks of loblolly (Pinus taeda L.) and slash pine (P. elliottii Engelm. var. elliotttii). Efficient operational deployment of genetic materials requires an understanding of possible site and silvicultural treatment interactions to maximize yield potential. We examined genotype (family) by environmental interactions (G × E) through age 5 years using a factorial experiment consisting of silvicultural treatment intensity (operational versus intensive), planting density (1334 versus 2990 trees ha⁻¹) and families (seven elite full-sib loblolly and six elite full-sib slash pine families). In January of 2000, randomized complete block, split-plot experiments were installed at two locations for each species in southeast Georgia and northeast Florida. Five years after planting, both loblolly and slash pine demonstrated significant interactions among several factors: genotype × location (p < 0.028 and p < 0.016, respectively), genotype × silvicultural treatment intensity (p < 0.055 and p < 0.059), and silvicultural treatment intensity × density (p < 0.002 and p < 0.001) for basal area (BA) and standing stem volume (VOL). Genotype × silvicultural treatment interactions were positive, with the best overall performing families responding the greatest to intensive treatment. There were changes in slash pine family rankings between locations, which were partly explained by reductions in growth associated with a combination of fusiform rust infection [Cronartium quercum (Berk.) Miyabe ex Shirai f. sp. fusiforme] and wind damage from the 2004 hurricane season. No three-way interactions, which included family, were evident and all genetic sources were stable across the contrasting planting densities. At age 5, loblolly pine outperformed slash pine (p < 0.0001), especially under the intensive silvicultural intensity. While loblolly performance was similar whether deployed in mixtures or pure family blocks, slash pine tended to be more productive in intimate mixtures than when grown in pure family blocks (p = 0.0754 for aboveground biomass).     

Can intensive management accelerate the restoration of Brazil's Atlantic forests?

Only 7% of the once extensive forest along the eastern coast of Brazil remains, and much of that is degraded and threatened by agricultural expansion and urbanization. We wondered if methods similar to those developed to establish fast-growing Eucalyptus plantations might also work to enhance survival and growth of rainforest species on degraded pastures composed of highly competitive C₄ grasses. An 8-factor experiment was laid out to contrast the value of different intensities of cultivation, application of fertilizer and weed control on the growth and survival of a mixture of 20 rainforest species planted at two densities: 3 m × 1 m, and 3 m × 2 m. Intensive management increased seedling survival from 90% to 98%, stemwood production and leaf area index (LAI) by ～4-fold, and stemwood production per unit of light absorbed by 30%. Annual growth in stem biomass was closely related to LAI alone (r² = 0.93, p < 0.0001), and the regression improved further in combination with canopy nitrogen content (r² = 0.99, p < 0.0001). Intensive management resulted in a nearly closed forest canopy in less than 4 years, and offers a practical means to establish functional forests on abandoned agricultural land.     

Utilization and traditional strategies of in situ conservation of iroko (Milicia excelsa Welw. C.C. Berg) in Benin

Socio-cultural surveys were carried out on the basis of a questionnaire administered on 346 respondents in order to investigate cultural and ethnobotanic uses of Milicia excelsa in Benin.M. excelsa contributes to cure 45 human diseases. The different parts of the tree used are leaves (30.3%), bark (25.8%), root (23.6%), latex (10.1%), flaking bark (6.7%), wood, calcium concentrated in old trees, and gum (1.1% each). Fruits or seeds are rarely used. Six different forms of utilization were recorded: soaking (46.3%), bark or leaves decoction (32.8%), herb tea (11.9%), powder (6.0%), leaves or bark grounded and rolled up into ball (1.5%), component of offering to fetish (1.5%). Iroko wood is also used in carpentry and joinery for construction purposes; furniture as well as for building boats/canoes.Iroko tree is used as the conservatory of cultural values and incarnates many divinities, which differ significantly from one province to another (χ² = 1830.27; d.f. = 25; P < 0.01%). There is a significant difference between the provinces in respect of the recognition of the species (χ² = 268.71; d.f. = 17; P ≤ 0.01%) and the population awareness about iroko as a sacred tree also varied from one province to another (χ² = 308.66; d.f. = 27; P ≤ 0.01%).Veneration of the tree is the main approach of its conservation by local people. M. excelsa is conserved on farm, in sacred groves, in public places and in cemeteries. The different sacred objects used to symbolize the divinities incarnated by iroko are: pottery (36.36%), iron (11.11%), calabash (4.04%), candle (2.02%), piece of cloth (18.18%), sacrifice (13.13%), piece of money (3.0%), stone (2.05%), glassware (broken bottle, 2.02%), and convent (8.08%). There is a highly significant difference between provinces as far as the sacred objects are concerned (χ² = 183.037; d.f. = 19 and P < 0.001%). The conservation purposes also vary significantly from one region to other (χ² = 894.47; d.f. = 31; P < 0.01%).     

Effects of rhinacanthin-C on function and expression of drug efflux transporters in Caco-2 cells

Rhinacanthin-C is a bioactive naphthoquinone ester found in Rhinacanthus nasutus Kurz (Acanthaceae). This compound has potential therapeutic value as an anticancer and antiviral agent. The purposes of this study were to determine the effects of this compound on the function and the expression of P-glycoprotein (P-gp) and multidrug resistance-associated protein 2 (MRP2), using the in vitro model of Caco-2 cells. The activities of P-gp and MRP2 were determined by following the intracellular accumulation of calcein and 5(6)-carboxy-2′,7′-dichlorofluorescein in the uptake assays with fluorescence spectroscopy. The expression of P-gp after prolonged exposure was evaluated by flow cytometry with the use of a fluorescein isothiocyanate-conjugated anti-human P-gp antibody. Our results showed that the inhibitory effect of rhinacanthin-C was more potent toward P-gp than MRP2, and was reversible. Short-term exposure of Caco-2 cells with rhinacanthin-C (100 μM) resulted in increase in P-gp expression without any significant change in its function. Extended exposure of Caco-2 cells to the naphthoquinone at the highest non-cytotoxic concentration (0.625 μM) for 7 days had no effect on the expression and the function of P-gp. These findings suggested that rhinacanthin-C might raise the problem of herb–drug interaction when co-administered with other P-gp substrates.     

Effects of globularifolin on cell survival,nuclear factor-κB activity,neopterin production,tryptophan breakdown and free radicals in vitro

The potential effects of globularifolin, an acylated iridoid glucoside, on cell survival, inflammation markers and free radicals scavenging were investigated. Viability assay on human myelomomonocytic cell line THP-1 and human peripheral blood mononuclear cells (PBMC) using the Cell-Titer Blue assay proved that globularifolin had no toxic effect at the tested concentrations. Conversely, it is proportional to the dose globularifolin increased growth of THP-1 cells (p < 0.01). On human PBMC, globularifolin at 6.25 and 12.5 μM concentrations showed a stimulatory effect, while at 12.5–200 μM it suppressed response of PBMC to stimulation with phytohemagglutinin (PHA). Globularifolin (50–200 μM) enhanced neopterin formation dose-dependently, whereas tryptophan breakdown was not influenced. At 50–200 μM in unstimulated PBMC in THP-1 cells, globularifolin induced a significant expression of nuclear factor-κB (NF-κB) as was quantified by Quanti-Blue assay. By contrast, in lipopolysaccharide (LPS)-stimulated cells, the higher concentrations of globularifolin suppressed NF-κB expression dose-dependently and a significant decrease was observed at 200 μM concentration. A positive correlation was found between increased neopterin and NF-κB activity (p < 0.01). Similarly, a positive correlation was observed between neopterin levels in mitogen-induced cells and NF-κB activity in LPS-stimulated cells after treatment with globularifolin (p = 0.001). The free radical scavenging capacity of globularifolin evaluated by Oxygen Radical Absorbance Capacity (ORAC) assay showed relative ORAC values of 0.36 ± 0.05 μmol Trolox equivalent/μmol. All together, results show that natural antioxidant globularifolin might represent a potential immunomodulatory as well as proliferative agent, which deserves further in vitro and in vivo studies.     

Fine-root distribution and morphology in an acidic Norway spruce (Picea abies (L.) Karst.) stand in SW Sweden in relation to granulated wood ash application

Wood ash is recommended as a compensatory fertiliser to counteract the effects of acidic deposition on forest ecosystems. Spatial distribution of biomass, necromass and morphology parameters of the fine roots (diameter classes <1, 1–2, <2 mm) of Norway spruce (Picea abies (L.) Karst.) were analysed in response to fertilisation with granulated wood ash (GWA) in a long-term field experiment in SW Sweden. GWA was applied as a single dose of 3200 kg ha⁻¹ and the fine roots were sampled 9 years later by soil coring. Soil cores were divided into 1-cm strata within the top 0–2.5 cm humus limits, the lower humus below 2.5 cm (with varying thickness) and the mineral soil to 50 cm depth (from ground surface). Total fine-root biomass in the control (C) and GWA treatment, 256 ± 20 and 258 ± 25 g m⁻², respectively, and length 2072 ± 182 and 1800 ± 198 m m⁻², respectively, did not differ statistically from each other. Total fine-root necromass in the 1–2 mm fraction was significantly higher in C than in the GWA treatment, 130 ± 12 and 80 ± 10 g m⁻², respectively. Fine-root biomass in the <1 mm fraction was significantly greater in the lower humus in the GWA treatment, but this did not affect the total biomass in the <1 mm fraction in the whole soil profile. The biomass-to-necromass ratio (1–2 mm) was significantly higher in the GWA treatment in the 0–30 cm soil layer than in the corresponding layer of the control. Specific root length (SRL) was lower in the GWA treatment than in the control in the 0–5 cm soil layer. The lower necromass and SRL were more clearly related to the GWA treatment, whereas the difference in the vertical distribution of biomass may have been related to the thicker humus layer in the GWA plots.     

Tree and forest encroachment into fescue grasslands on the Cypress Hills plateau,southeast Alberta,Canada

Tree encroachment into rough fescue (Festuca campestris) grassland has been identified as an ecological concern on the Cypress Hills plateau in southeastern Alberta, Canada. A combination of field sampling (109 transects), a dendrochronological assessment (1361 trees), and a time series analysis of remotely sensed images from five different time periods (1950–2002) were used to determine the extent and rate of tree encroachment and forest development. Tree cover increased by 768 ha (～51%) between 1950 and 2007, representing 10% of the study area, from 1502 ha of pre-1950 forest. Post-1950 tree invasion also created an ingress zone of 750 ha (～10% of study area) based on field transects. Forest cover increased at a consistent rate of 14.3 ha/year. Lodgepole pine (Pinus contorta var. latifolia) was the most common tree species associated with encroachment. Invasion based on the number of established trees occurred at an exponential rate of 3.1%/year after 1890, with density increased by filling spaces adjacent to and between trees within the grassland vegetation. The rate of tree establishment increased to 4.4%/year after 1980, suggesting a change occurred in environmental conditions. Annual atmospheric temperatures increased 0.55 °C from 1929 to 2005 (P < 0.001), with winter (December–February) and spring (March–May) temperatures accounting for most of the increase (P < 0.001), whereas summer (June–August) temperatures slightly decreased (P < 0.050, 0.34 °C) and precipitation increased (P < 0.005, 30 mm). Cattle and elk (Cervus elaphus) grazing was not considered a primary factor for explaining tree encroachment. Based on multidimensional scaling, lodgepole pine establishment was associated with warmer spring temperatures and greater fall (October–November) precipitation. A landscape devoid of wildfires, combined with greater moisture availability, and a longer frost-free season is likely conducive to the sustained establishment of coniferous trees and forest development within the Cypress Hills fescue grassland ecosystem. Long-term conservation of the fescue grasslands could be possible by reintroducing fire.     

Stand development after different thinnings in two uneven-aged Picea abies forests in Sweden

Two field experiments, located in Central and Northern Sweden, were used to study the influence of standing volume on volume increment and ingrowth in uneven-aged Norway spruce (Picea abies (L.) Karst.) stands subjected to different thinnings. Each experiment had a 3 × 2 factorial block design with two replications. Treatments were thinning grade, removing about 45, 65, and 85% of pre-thinning basal area, and thinning type, removing the larger or the smaller trees, respectively. Each site also had two untreated control plots. Plot size was 0.25 ha. Volume increment was 0.5–6.8 m³ ha⁻¹ year⁻¹ for the plots, and significantly positively (p < 0.01) correlated with standing volume. Within treatment pairs, plots thinned from Above had consistently higher volume increment than plots thinned from Below. Ingrowth ranged from 3 to 33 stems ha⁻¹ year⁻¹, with an average of 14 and 21 stems ha⁻¹ year⁻¹ at the northern and southern site, respectively. At the southern site ingrowth was significantly negatively (p < 0.01) correlated with standing volume, but not at the northern site. Mean annual mortality after thinning was 2 and 7 stems ha⁻¹ year⁻¹at the northern and southern site, respectively.     

Pharmacokinetic and metabolism studies of rohitukine in rats by high performance liquid-chromatography with tandem mass spectrometry

A sensitive, selective, and rapid high performance liquid chromatography–tandem mass spectrometry (LC–MS/MS) was developed for the quantification of rohitukine in rat plasma. HPLC was performed using a Symmetry-Shield C₁₈ (5 μ, 4.6 × 150 mm) column, and isocratic elution with ammonium acetate buffer (pH 4; 10 mM):methanol (08:92, v/v) at a flow rate of 0.6 mL/min. Sample clean-up involved solid phase extraction (SPE) of analyte and internal standard (phenacetin) from 100 μL plasma. The parent → product ion transitions (MRM) for analyte and IS were 306.1 → 245.1 m/z and 180.1 → 138.1 m/z respectively, and were monitored on a triple quadrupole mass spectrometer, operating in positive ion mode. The method was validated across the dynamic concentration range of 5–500 ng/mL for rohitukine, with a fast run time of 4.5 min. The analytical method measured concentrations of rohitukine with accuracy (% bias) of <± 10% and precision (% RSD) of <± 12%. Rohitukine was stable during the battery of stability studies viz., bench-top, auto-sampler, freeze/thaw cycles and 30 days of storage in a freezer at − 70 ± 10 °C. Finally, the applicability of this assay has been successfully demonstrated in vivo pharmacokinetic and in vitro metabolism studies in Sprague–Dawley rat. This method will therefore be highly useful for future preclinical and clinical pharmacokinetic studies of rohitukine.     

Population structure and nut yield of a Bertholletia excelsa stand in Southwestern Amazonia

Although Brazil nut (B. excelsa) is often touted as one of the most economically successful NTFPs, little is known about the population structure of this species within its natural range in Southwestern Amazonia or ecological factors that affect fruit production. Since these are considered fundamental for sustainable resource management, we examined a natural Brazil nut stand in an extractive reserve in Acre, Brazil, posing the following questions: (1) What is the spatial distribution, species density, and size–class structure of B. excelsa? and (2) What tree-level factors influence Brazil nut production? In a 420 ha census, 568 trees ≥10 cm diameter at breast height (dbh) were counted, resulting in a density of 1.35 trees ha⁻¹. Based on the nearest-neighbor method, an index of aggregation (R) of 0.77 indicated a rejection of the null hypothesis of a strictly random distribution pattern. Yet, this value suggests a much greater tendency toward randomness than either clumping or uniformity. Our data do not show the commonly reported existence of groves, referring to clearly defined clusters of 50 to several hundred trees separated from similar clusters by great distances. Almost 1/4 of the population (23%) was composed of non-reproductive juveniles. Maximum R² improvement analysis applied to four distinct diameter classes provided insight into the dynamics of production-related variables over the species life cycle. While dbh explained 1/3 of production variance (R² = 0.3360) in the smallest diameter class (10 cm ≤ dbh < 50 cm), which included those in the process of reaching reproductive maturity, crown form best explained production variance of very large trees (dbh ≥ 100 cm). Results also demonstrated a significant negative correlation between crown vine load and production of trees ≥ 50 cm dbh (r = −0.13, P = 0.008), suggesting the need for further study on vine cutting as a possible silvicultural treatment for enhancing nut yields.     

New cytotoxic triterpenoid saponins from the whole plant of Clematis lasiandra Maxim

Four new oleanane type triterpenoid saponins (1–4) and three known saponins (5–7) were isolated from the whole plant of Clematis lasiandra Maxim. The structures of the four new compounds were elucidated as 3-O-β-d-ribopyranosyl-(1 → 3)-α-l-rhamnopyranosyl-(1 → 2)-[β-d-glucopyranosyl-(1 → 4)]-β-d-xylopyranosyl hederagenin (1), 3-O-β-d-ribopyranosyl-(1 → 3)-α-l-rhamnopyranosyl-(1 → 2)-β-d-xylopyranosyl oleanolic acid 28-O-β-d-glucopyranosyl ester (2), 3-O-β-d-ribopyranosyl-(1 → 3)-α-l-rhamnopyranosyl-(1 → 2)-β-d-xylopyranosyl hederagenin (3) and 3-O-β-d-ribopyranosyl-(1 → 3)-α-l-rhamnopyranosyl-(1 → 2)-[β-d-glucopyranosyl-(1 → 4)]-α-l-arabinopyranosyl hederagenin (4) on the basis of extensive spectroscopic analysis and chemical evidence. Compounds 1–7 were evaluated for their cytotoxicity against human tumor cell lines HL-60, Hep-G2 and SGC-7901, and all of the evaluated saponins showed significant cytotoxicity to those three tumor cell lines with IC₅₀ in the range from 1.40 to 19.50 μmol/L except for compounds 2 and 6.     

Initial impact of supervised logging and pre-logging climber cutting compared with conventional logging in a dipterocarp rainforest in Sabah,Malaysia

Pre-marked skid trails, directional felling and climber cutting when logging in tropical rainforests may be important ways of reducing damage to the forest, thus creating a healthier stand and improving future yields.This study, carried out in a virgin dipterocarp rainforest in the south of Sabah, Malaysia, compared two types of logging (both with and without pre-cutting climbers): conventional selective logging (CL) and supervised logging (SL). The latter is a selective logging system in which both pre-marked skid trails and directional felling were implemented. The pre-marked skid trails were aligned parallel to each other, spaced 62 m apart. A randomised complete block 2 × 2 factorial design was used in the experiment, consisting of 16 gross treatment plots, each of 5.76 ha with a 1 ha net plot in the centre.Fewer trees tended (0.050 < P ≤ 0.100) to be logged in SL plots than in CL plots (on average 9.4 and 13.0 trees ≥60 cm diameter breast height ha⁻¹). Pre-felling of climbers resulted in four more dipterocarp trees being logged ha⁻¹, compared with no climber cutting: a statistically significant difference (P ≤ 0.050). The basal areas lost of both large trees (≥ 60 cm dbh) and small dipterocarp trees (10–29 cm dbh) tended to differ between the logging systems, with CL leading to greater losses.There were significant differences in the residual stands left by the logging systems, with respect to the number of dipterocarps and their basal area in the diameter class 10–29 cm; ca 30% more stems being found after SL. No significant differences (or tendencies) in these variables were found in the residual stands in other diameter classes, or when trees of all species were considered.     

Lactones from Ligusticum chuanxiong Hort. reduces atherosclerotic lesions in apoE-deficient mice via inhibiting over expression of NF-kB -dependent adhesion molecules

The present study aims to investigate the anti-atherosclerotic effects of lactones extracted from Ligusticum chuanxiong Hort (LLC) in apoE-deficient mice (ApoE^−/− mice) and proclaim its underlying mechanisms. Expression of endothelial adhesion molecules and NF-κB around the atherosclerotic lesions was detected by immunohistochemistry (IHC). To further validate the mechanism, effect of LLC on the secretion of ICAM-1 and VCAM-1 of human umbilical vein endothelial cells (HUVECs) induced by tumor necrosis factor α (TNF-α) was measured by ELISA. And the activation of NF-κB was detected by western blot. Mice treated with LLC showed significant reduction in lesion sizes of thoracic segments of the aorta (p < 0.01). Meanwhile, LLC treatments lead to decreases of serum TG, TC and LDL-C contents, respectively. LLC also decreased the expression of CD31, intercellular adhesion molecule-1 (ICAM-1), monocyte chemoattractant protein-1 (MCP-1) and nuclear factor-kappa B (NF-κB) in the atherosclerotic plaque. Moreover, LLC at 3.125–25 μg/mL can dose-dependently attenuate the expression of ICAM-1 and VCAM-1 in TNF-α stimulated HUVECs. Western blot result indicated LLC inhibited activation of NF-κB. These results suggested that LLC could ameliorate atherosclerosis in ApoE^−/− mice. The mechanism of action of LLC on anti-atherosclerotic effect may be attributed to the suppression of the production of NF-κB-dependent adhesion molecules.     

Pharmacokinetic study of eight coumarins of Radix Angelicae Dahuricae in rats by gas chromatography–mass spectrometry

A sensitive, specific gas chromatography–mass spectrometry (GC–MS) method was established for the quantitative determination of eight coumarins of Radix Angelicae Dahuricae including coumarin, isopsoralen, psoralen, xanthotoxin, bergapten, osthole, imperatorin and oxypeucedanin in rat plasma. Nitrendipine was used as the internal standard (IS). The plasma samples were extracted by acetonitrile. GC separation was accomplished on a DB-5MS column with temperature programmed from 160 °C (17 min) to 190 °C (10 min) at the rate of 20 °C/min, then to 240 °C (5 min) at 20 °C/min, and finally to 280 °C (14 min) at 20 °C/min. The detection was performed on a quadrupole mass spectrometer detector operated by full-scan mode (m/z 50–500). The lower limit of quantitation was 5–10 ng/mL for eight coumarins, and the linear range was 5–1000 ng/mL for the coumarins (R² > 0.9990). All the validation data were within the required limits. After oral administration, the plasma concentration–time curves showed that the time for maximum concentration (T_max) was 1.29 for coumarin, 1.83 for isopsoralen, 1.93 for psoralen, 1.30 for xanthotoxin, 2.04 for bergapten, 0.64 for osthole, 1.41 for imperatorin and 0.51 h for oxypeucedanin. The plasma concentration of the eight coumarins was low, with a mean maximum plasma concentration (C_max) < 6.41 μg/mL. Pharmacokinetic data analysis showed that the eight coumarins had different pharmacokinetic characteristics after oral administration. The method was successfully applied to pharmacokinetic studies of eight coumarins after oral administration in rats.     

Mineral nitrogen dynamics following soil compaction and cultivation during hoop pine plantation establishment

The effects of soil compaction and cultivation on soil mineral N dynamics were investigated through an 18-month, in situ N mineralisation experiment during the inter-rotation and early establishment period of a second rotation (2R) hoop pine (Araucaria cunninghamii Aiton ex A. Cunn) plantation in southeast Queensland, Australia. Treatments were 0, 1 and 16 passes of a fully laden forwarder (gross weight, 40.2 Mg) and cultivation by disc plough (zero cultivation and cultivation). Nitrate N was the dominant form of mineral N throughout the 18-month sampling period in both non-cultivated and cultivated soils, varying between 10 and 40 kg ha⁻¹ whilst ammonium N remained <10 kg ha⁻¹. Compaction had no significant effect on N mineralisation or nitrification. However, the remediation of the effects of compaction on soil through the use of the disc plough had significant impacts on N mineralisation, nitrification and N leaching. On a seasonal basis, the mean net N mineralisation increased from around 30 to 53 kg ha⁻¹, nitrification from 28 to 43 kg ha⁻¹ and nitrate N leaching from around 10 to 73 kg ha⁻¹ following cultivation.     

Baicalein decreases side population proportion via inhibition of ABCG2 in multiple myeloma cell line RPMI 8226 in vitro

ObjectiveTo investigate the effect of baicalein on side population in human multiple myeloma cell line RPMI 8226 and the underlying molecular mechanisms in vitro and in silico.MethodsMTT assay was applied to detect the anti-proliferation effect of baicalein. The detection of side population cells is based on the Hoechst 33342 exclusion assay technique and flow cytometric analysis. Western blotting assay was used to explore the expression of ABCG2 protein. Homology modeling and molecular docking were performed with Discovery Studio 2.1.ResultsBaicalein decreased both cell viability with IC₅₀ = 168.5 μM and the proportion of SP cells in a dose-dependent manner. Correspondingly, it significantly decreased the expression level of ABCG2 protein. Baicalein also shared similar binding sites and modes with fumitremorgin C to the protein.ConclusionsBaicalein possessed novel anticancer properties, such as anti-proliferation and drug efflux inhibition in side population cells, which suggested its potential feature of targeting cancer stem cells of multiple myeloma.