Individual and combined effects of rhynchophylline and ketamine on proliferation,NMDAR1 and GluA2/3 protein expression in PC12 cells

Rhynchophylline is an active component of the Uncaria species, which is a member of the Rubiaceae family. Our studies show that the downregulation of N-methyl-d-aspartate (NMDA) receptor subunit GluN2B expression in the nucleus accumbens, amygdala, medial prefrontal cortex, and hippocampal CA1 area by rhynchophylline is beneficial for the treatment of psychological dependence on amphetamines. The individual and combined effects of rhynchophylline and ketamine on proliferation and GluN1 and GluA2/3 protein expression in PC12 cells were investigated. PC12 cells were differentiated into neuron-like cells by treatment with nerve growth factor (50 ng/mL). After treatment for 48 h, differentiated PC12 cell proliferation and GluN1 and GluA2/3 protein expression were analyzed. The viability of PC12 cells was reduced by ketamine at doses of 0.50, 1.00, 1.50, and 2.00 mmol/L, with the viability of cells treated with 1.50 and 2.00 mmol/L of ketamine significantly lower than that of the control cells. However, PC12 cells treated with rhynchophylline showed no toxicity at doses of 0.25, 0.50, 0.75, or 1.00 mmol/L. While GluA2/3 protein expression was upregulated by ketamine, it was not influenced by rhynchophylline. GluN1 protein expression was downregulated by rhynchophylline (1 mmol/L), while treatment with ketamine, either alone or with rhynchophylline, had no effect. These findings demonstrate that rhynchophylline suppresses GluA2/3 expression in ketamine-induced PC12 cells and downregulates GluN1 expression. Ketamine's lack of effect on GluN1 expression offers a partial explanation for ketamine addiction and the anti-addictive properties of rhynchophylline.     

Phytic acid enhances the oral absorption of isorhamnetin,quercetin, and kaempferol in total flavones of Hippophae rhamnoides L.

AimTotal flavones of Hippophae rhamnoides L. (TFH) have a clinical use in the treatment of cardiac disease. The pharmacological effects of TFH are attributed to its major flavonoid components, isorhamnetin, kaempferol, and quercetin. However, poor oral bioavailability of these flavonoids limits the clinical applications of TFH. This study explores phytic acid (IP₆) enhancement of the oral absorption in rats of isorhamnetin, kaempferol, and quercetin in TFH.MethodsIn vitro Caco-2 cell experiments and in vivo pharmacokinetic studies were performed to investigate the effects of IP₆. The aqueous solubility and lipophilicity of isorhamnetin, quercetin, and kaempferol were determined with and without IP₆, and mucosal epithelial damage resulting from IP₆ addition was evaluated by MTT assays and morphology observations.ResultsThe P_app of isorhamnetin, kaempferol, and quercetin was improved 2.03-, 1.69-, and 2.11-fold in the presence of 333 μg/mL of IP₆, respectively. Water solubility was increased 22.75-, 15.15-, and 12.86-fold for isorhamnetin, kaempferol, and quercetin, respectively, in the presence of 20 mg/mL IP₆. The lipophilicity of the three flavonoids was slightly decreased, but their hydrophilicity was increased after the addition of IP₆ in the water phase as the logP values of isorhamnetin, kaempferol, and quercetin decreased from 2.38 ± 0.12 to 1.64 ± 0.02, from 2.57 ± 0.20 to 2.01 ± 0.04, and from 2.39 ± 0.12 to 1.15 ± 0.01, respectively. The absorption enhancement ratios were 3.21 for isorhamnetin, 2.98 for kaempferol, and 1.64 for quercetin with co-administration of IP₆ (200 mg/kg) in rats. In addition, IP₆ (200 mg/kg, oral) caused neither significant irritation to the rat intestines nor cytotoxicity (400 μg/mL) in Caco-2 cells.ConclusionsThe oral bioavailability of isorhamnetin, kaempferol, and quercetin in TFH was enhanced by the co-administration of IP₆. The main mechanisms are related to their enhanced aqueous solubility and permeability in the presence of IP₆. In summary, IP₆ is a potential absorption enhancer for pharmaceutical formulations that is both effective and safe.     

Identification of chicoric acid as a hypoglycemic agent from Ocimum gratissimum leaf extract in a biomonitoring in vivo study

Ocimum gratissimum L. is popularly used to treat diabetes mellitus. The hypoglycemic activity of this medicinal species has been confirmed by in vivo studies. The present study conducted a chemical investigation of a leaf decoction (10% p/v) of O. gratissimum monitored by in vivo hypoglycemic activity assays. Four phenolic substances were identified: l-caftaric acid (1), l-chicoric acid (2), eugenyl-β-d-glucopyranoside (3) and vicenin-2 (4). The acute hypoglycemic activity of the O. gratissimum decoction fractions Og1-S (300 mg/kg), Og1-A (240 mg/kg) and Og1-B (80 mg/kg) was evaluated intraperitoneally in normal and streptozotocin-induced diabetic mice. They reduced glycemia by 63%, 76% and 60% (in 120 min), respectively, in the diabetic mice. Subfractions of Og1-A were also evaluated under the same conditions: Og1-AS (200 mg/kg) and Og1-AP (40 mg/kg) produced a decrease of only 37% and 39%, respectively. Among the major phenolic substances, only chicoric acid (2; 3 mg/kg) reduced significantly the glycemic levels of diabetic mice by 53%, 120 min after treatment. This is the first study describing the hypoglycemic activity of chicoric acid in an animal model of diabetes mellitus. In addition, we suggest that there may be other substances contributing to this activity. Thus, for the first time, a correlation is established between the hypoglycemic activity of O. gratissimum and its chemical composition.     

Isoflavone content and estrogenic activity of different batches of red clover (Trifolium pratense L.) extracts: An in vitro study in MCF-7 cells

The estrogenicity of different batches of red clover (Trifolium pratense L., Fabaceae; RCL) extracts and its relationship with the isoflavone content were assessed by measuring MCF-7 cell proliferation by flow cytometry and propidium iodide staining. RCL extracts were compared to estradiol (E2) and to the main RCL isoflavones biochanin A, daidzein, genistein and formononetin. Isoflavone content in the extracts was assayed by HPLC.E2 and isoflavones increased MCF-7 proliferation in a concentration-dependent fashion, with the following potency order: E2 >>> genistein > biochanin A = daidzein > formononetin. Extracts increased MCF-7 proliferation with different potencies, which in four out of five extracts correlated with the ratios 5,7-dihydroxyisoflavones/7-hydroxyisoflavones. The efficacy of all extracts increased with decreasing genistein contents. A solution containing the main isoflavones at the average concentration of RCL extracts increased MCF-7 proliferation with higher potency and steeper concentration–response curve. The effects of E2, of RCL extracts and of the isoflavone solution were inhibited by the estrogen receptor antagonist 4-hydroxytamoxifen.Flow cytometric analysis of MCF-7 proliferation is a suitable bioassay for the estrogenicity of RCL extracts, thus expanding the characterization of individual batches beyond assessment of chemical composition and contributing to improved standardization of quality and activity.     

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.     

Influence of various in situ rainwater harvesting methods on soil moisture and growth of Tamarix ramosissima in the semiarid loess region of China

The influence of different in situ rainwater harvesting and moisture conservation methods on soil moisture storage and growth of Tamarix ramosissima was studied in the semiarid loess region of China from 2002 to 2004. The treatments included control (T1), trench (T2), saucer covered by plastic film (T3), bare ridge and bare furrow (T4), plastic-covered ridge and bare furrow (T5), and plastic-covered ridge and gravel-covered furrow (T6). The results indicated that soil water storage for the T5 and T6 was significantly higher than the control. The T6 treatments produced the highest amount of soil water storage, 18–137 mm more than the controls at soil depth of 0–100 cm and 40–75 mm at soil depth of 100–200 cm. No significant differences in soil water storage were found between the T2, T3, T4 and the control treatments at soil depth of 0–100 cm, but soil water storage at soil depth of 100–200 cm was significantly higher for the T2, T3, T4 treatment than the control.Rainwater harvesting and moisture conservation treatments increased growth of T. ramosissima, tree height was significantly higher for the rainwater harvesting and moisture conservation treatments than the controls. Tree height, crown diameter and collar girth for the T6 treatments increased by 70, 57 and 79% as compared to the controls. No significant differences in crown diameter and collar girth were observed between T1, T2 and T3 or T4 and T5 treatments in some years. The effectiveness of the different treatments for the tree height, crown diameter and collar girth was in the order of T6, T5, T4, T3, T2 and T1.     

Nitrate concentrations in soil solutions below Danish forests

Nitrate in the soil water below the root zone is a pre-condition for nitrate leaching, and it indicates loss of nutrients from the forest ecosystem. Nitrate leaching may potentially cause eutrophication of surface water and contamination of ground water. In order to evaluate the extent of nitrate leaching in relation to land-use, a national monitoring programme has established sampling routines in a 7×7 km grid including 111 points in forests. During winters of 1986–1993, soil samples were obtained from a depth of 0–25, 25–50, 50–75 and 75–100 cm. Nitrate concentrations in soil solutions were determined by means of a 1 M KCl extraction. The influence of forest size, forest-type, soil-type, tree species and sampling time on the nitrate concentrations was analysed in a statistical model. The analysis focused on data from depth 75–100 cm, as nitrate is considered potentially lost from the ecosystem at this depth. The range of nitrate concentrations was 0–141 mg NO⁻₃–N dm⁻³ and the estimated mean value was 1.51 mg NO⁻₃–N dm⁻³. The concentration was influenced by (1) forest size (concentrations in forests <10 ha were higher than concentrations in forests >50 ha), (2) forest-type (afforested arable land had higher concentrations than forest-type `other woodland'), (3) soil-type (humus soils showed above average concentrations, and fine textured soils had higher concentrations than coarse textured soils), and (4) sampling time. Unlike other investigations, there was no significant effect of tree species. A few sites deviated radically from the general pattern of low concentrations. The elevated concentrations recorded there were probably caused by high levels of N deposition due to emission from local sources or temporal disruptions of the N cycle. The nitrate concentration in the soil solution below the root zone was mostly rather low, indicating that, generally, N saturation has not yet occurred in Danish forest ecosystems. However, median concentrations exceeding drinking water standards (11.3 mg NO⁻₃–N dm⁻³) were found at 7% of the sites. Furthermore, 30% of the sites had median concentrations above 2 mg NO⁻₃–N dm⁻³, suggested as an elevated level for Danish forest ecosystems, equalling annual N losses of more than 2–6 kg ha⁻¹ year⁻¹.     

Looking beyond sugars: Phytochemical profiling and standardization of manna exudates from Sicilian Fraxinus excelsior L.

Different grades of genuine and counterfeit Fraxinus excelsior exudates, marketed as natural sweeteners or mild laxatives, were evaluated for their proximate composition and for saccharidic, organic acids, lipidic and phenolic profile by means of GC–MS and ¹H NMR. Genuine samples contained mannitol (39–48 g/100 g, according to the grade), fructose (9–16 g/100 g), glucose (2–3.7 g/100 g), sorbitol (0,5–0,6 g/100 g), galactose (0.02–0.74 g/100 g), oligosaccharides as mannotriose (13–22 g/100 g) and stachyose (1–11 g/100 g), and traces of myo-inositol, mannose, sucrose. On the contrary, counterfeit samples contained mostly mannitol and sorbitol, with traces of fructose, glucose and mannose. Differences in ash, total polyphenolic content and fatty acid composition allowed a quick identification of counterfeit products, confirmed by a distinct mono-, oligosaccharidic and phenolic pattern. Elenolic acid (63–1628 mg/kg), tyrosol (15–774 mg/kg), homovanillic acid (2,39–52.8 mg/Kg), dopaol (0.8–63 mg/kg), pinoresinol (4.2–18.5 mg/kg) and fraxetin (0.25–11.64 mg/kg), albeit showing a wide concentration range, were the most abundant substances detected in the phenolic fraction of Fraxinus manna, while esculetin, p-hydroxybenzoic acid, 4-hydroxyphenacetic acid, 3,4 hydroxybenzoic acid, hydroxy-pinoresinol, medioresinol and siringaresinol were present in low amounts. The polyphenolic profile may be used as a marker for authentication and should be considered in the evaluation of nutritional and health properties ascribed to Fraxinus manna.     

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.     

Changes in soil,seepage water and needle chemistry between 1984 and 2004 after liming an N-saturated Norway spruce stand at the Höglwald,Germany

In 1984, a liming experiment with a surface application of 4 t ha⁻¹ of dolomitic limestone was started at the acidic N-saturated Norway spruce forest “Höglwald” in southern Germany and monitored until 2004. The decay of surface humus due to the accelerated mineralisation accounted for 18.5 ± 2.7 t ha⁻¹ C or 50% of the initial pool and 721.6 ± 115.0 kg ha⁻¹ N or 46% for N. Due to some translocation of organic material to the mineral soil the values to 40 cm depth are slightly lower (13.5 ± 4.4 t ha⁻¹ C or 15% of the initial pool and 631.6 ± 192.8 kg ha⁻¹ N or 13% for N). In the control plot NO₃⁻ concentrations at 40 cm depth were above the European level of drinking water (0.8 mmolc l⁻¹ or 50 mg NO₃⁻ l⁻¹) for nearly the whole investigation period. Liming increased NO₃⁻ concentrations in seepage water for approximately 15 years, and accelerated leaching losses by 396.2 NO₃⁻–N kg ha⁻¹ from 1984 to 2003. The increase in pH of the soil matrix was more or less restricted to the humus layer and the upper 5 cm of the mineral soil during the whole time span, while the base cations Ca and Mg reached deeper horizons with seepage water. From 1984 to 2003, an amount that nearly equalled the applied Mg, was leached out of the main rooting zone, while most of the applied Ca was retained. The time series of the elemental concentrations in needles showed minor changes. Ca concentrations in needles increased with liming, while Mg remained nearly unchanged, and P decreased in older needles.     

Two new steroidal saponins from Selaginella uncinata (Desv.) Spring and their protective effect against anoxia

Four steroidal saponins were isolated from the anti-anoxic fraction of the 60% EtOH extract of Selaginella uncinata, including two new compounds, (3β, 7β, 12β, 25R)-spirost-5-ene-3, 7, 12-triol-3-O-α-L-rhamnopyranosyl-(1 → 2)-O-[α-L-rhamnopyranosyl-(1 → 4)]-O-β-d-glucopyranoside (1), (2α, 3β, 12β, 25R)-spirost-5-ene-2, 3, 12-triol-3-O-α-L-rhamnopyranosyl-(1 → 2)-O-[α-L-rhamnopyranosyl-(1 → 4)]-O-β-d-glucopyranoside (2) and two known compounds, (3β, 12β, 25R)-spirost-5-ene-3,12-diol-3-O-α-L-rhamnopyranosyl-(1 → 2)-O-[α-L-rhamnopyranosyl-(1 → 4)]-O-β-d-glucopyranoside, (3), (1α, 3β, 25R)-spirost-5-ene-2-diol-3-O-α-L-rhamnopyranosyl-(1 → 2)-O-[α-L-rhamnopyranosyl(1 → 4)]-O-β-d-glucopyranoside (4). The four compounds showed potent protective effect against anoxia in the anoxic PC12 cells assay, among which compounds 1 and 2 were the most active. To our knowledge, this is the first study to report the steroidal saponins in the plant S. uncinata and demonstrate their protective effect against anoxia in PC12 cell assay.     

Effects of the extracts and an active compound curcumenone isolated from Curcuma zedoaria rhizomes on alcohol-induced drunkenness in mice

The Curcuma zedoaria rhizome has been used traditionally to treat gastrointestinal diseases as an aromatic stomachic drug, and this is currently used to treat alcohol-induced loss of appetite and nausea in Japan. We examined the effects of various fractions and isolated compounds on alcohol-induced drunkenness and blood alcohol concentrations in mice. The 30% ethanol-extract (1000 mg/kg) of C. zedoaria rhizome prevented drunkenness 60 and 120 min after 40% alcohol administration. The n-hexane-soluble fraction (300 mg/kg) and an isolated compound (3, 10 or 30 mg/kg) prevented drunkenness at 30, 60 or 120 min. The extract, n-hexane-soluble fraction and isolated compound reduced the elevation in blood alcohol concentrations 30 and 60 min after 40% alcohol administration. The isolated compound (10 and 30 mg/kg) enhanced liver ADH activity 30 and 60 min after 40% alcohol administration. The compound was identified as curcumenone by a direct comparison of ¹H- and ¹³C-NMR spectral data. In conclusion, the protective effect of the C. zedoaria extract on drunkenness might be due to an active substance, curcumenone, and decreases in the elevation of blood alcohol concentrations through increased liver alcohol dehydrogenase activity.     

Screening of osteoanagenesis-active compounds from Scutellaria baicalensis Georgi by hPDLC/CMC–online-HPLC/MS

In present study, an online analytical method using human periodontal ligament cell/cell membrane chromatography (hPDLC/CMC) combined with high-performance liquid chromatography/mass spectrometry (HPLC/MS) was used for direct recognition, separation, and identification of compounds for the first time from Scutellaria baicalensis Georgi (SBG) that are active on hPDLCs. Baicalein (BAI) and wogonin (WOG), which were identified as the active compounds of the ethyl ether extract of S. baicalensis Georgi (SBGEE), could bind to the same membrane receptor of hPDLC for simvastatin (SIM). Moreover, BAI (0.15–0.6 mg/L) and WOG (0.015–0.6 mg/L) had the capability to enhance cell proliferation, matrix calcification, and formation of calcified nodules, which are comparable to the activities of SIM (0.1 mg/L) in vitro. These observations are consistent with the K_A of the various drugs. It is very important for the development of SBG used to treat periodontitis.     

Coreosides A–D,C14-polyacetylene glycosides from the capitula of Coreopsis tinctoria and its anti-inflammatory activity against COX-2

Four new C₁₄-polyacetylene glycosides, namely coreosides A–D (1–4), were isolated from the capitula of Coreopsis tinctoria, a Snow chrysanthemum or Snow tea that is used as a folk tea for prevention of cardiovascular disease in southern Xinjiang, China. Coreosides A–D feature a long chain structure as its aglycon with two acetylenes on C-8 and C-10 and two olefinics on C-6 and C-12 sites, which construct a large conjugate system. The structures were elucidated on the basis of spectroscopic evidences and hydrolysis. Compounds 1–4 exhibited significant inhibition against cyclooxygenase-2 at the concentration of 1 × 10^− 6 mol/L, with its IC₅₀ values of 0.22–8.8 × 10^− 2 μmol/L.     

Carapanolides C–I from the seeds of andiroba (Carapa guianensis,Meliaceae)

Five new mexicanolide-type limonoids, carapanolides C–G (1–5), together with two new phragmalin-type limonoids, carapanolides H–I (6, 7), were isolated from the oil of Carapa guianasis AUBLET (Meliaceae) seeds. Their structures were elucidated on the basis of spectroscopic analyses using 1D and 2D NMR spectra and FABMS. Carapanolides C (1), E (3), and I (7) exhibited moderate activity in the P388 (IC₅₀ 17.9 μM in 1, 15.8 μM in 3) and L1210 cell lines (IC₅₀ 13.3 μM in 1, 18.1 μM in 3, 16.9 μM in 7). On the other hand, Carapanolide D (2) exhibited a strong inhibitory effect in the HL-60 cell line (IC₅₀ 11.0 μM), Carapanolides F (4) showed inhibitory activity in the L1210 cell line (IC₅₀ 15.9 μM), and the cytotoxic activity of Carapanolides I (7) was moderate in all cell lines.     

Life cycle inventories of roundwood production in northern Wisconsin: Inputs into an industrial forest carbon budget

Carbon budgets are developed to understand ecosystem dynamics and are increasingly being used to develop global change policy. Traditionally, forest carbon budgets have focused on the biological carbon cycle; however, it is important to include the industrial forest carbon cycle as well. The overall objective of this study was to quantify the major carbon fluxes associated with the production of Wisconsin's industrial roundwood, by using life cycle inventory (LCI) methodology to produce an industrial forest carbon budget. To achieve this objective we (1) developed carbon LCIs for the harvest process for three major forest ownerships (state, national, and private non-industrial), (2) developed carbon LCIs for a dimensional lumber and two oriented strand board (OSB) mills and (3) completed a scaled version of 1 and 2 to include more Wisconsin forestlands and to incorporate the other major processes within the industrial forest carbon cycle (e.g. primary mill, secondary mill, product use and product disposal processes of the industrial forest carbon cycle). The carbon budgets for the harvesting process of the Chequamegon-Nicolet National Forest (CNNF), the Northern Highland American Legion State Forest (NHAL), and the non-industrial private forests that participated in the managed forest laws of Wisconsin (MFL-NIPF) were 0.10, 0.18 and 0.11 tonnes C ha⁻¹ year⁻¹), respectively. The dimensional lumber and OSB products were both net carbon sources, and released 0.05–0.09 tonnes C/tonnes C processed). More carbon is sequestered than released within the industrial forest carbon cycle of Wisconsin's national (6 g C m⁻² year⁻¹), state (12 g C m⁻² year⁻¹) and non-industrial private forests (7 g C m⁻² year⁻¹). Using published net ecosystem production data we estimate that the net forest carbon cycle budget (sum of the biological and industrial C cycle, [Gower, S.T., 2003. Patterns and mechanisms of the forest carbon cycle. Ann. Rev. Environ. Resour. 28, 169–204]) for the CNNF ranges between −897 and 348 g C m⁻² year⁻¹. Life cycle inventories of wood and paper products should be clear and explicitly state what processes are included, so that results can be used by policy makers and future researchers.     

Optimization of the selective preparation of 20(R)-ginsenoside Rg3 catalyzed by d,l-tartaric acid using response surface methodology

The optimization of the selective preparation of 20(R)-ginsenoside Rg3 converting protopanaxadiol type saponins (PPD saponins) by the commercially available d, l-tartaric acid was carried out using response surface methodology (RSM) based on a three-factor and six-level central composite design. The optimal 20(R)-ginsenoside Rg3 de% was predicted to be 94.52% in the combination of the factors (d, l-tartaric acid concentration 1.19 mol/L, temperature 107.9 °C and time 2.79 h) through the canonical analysis with maximum responses. Under the optimum reaction conditions, the actual 20(R)-ginsenoside Rg3 de% was 96.49%. 20(R)-ginsenoside Rg3 (1) and 20(S)-ginsenoside Rg3 (2) were separated and identified by ¹H-NMR and ¹³C-NMR. Therefore, the RSM was effective to optimize the preparation of 20(R)-ginsenoside Rg3 by converting PPD saponins using d, l-tartaric acid.     

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.     

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.     

In vitro permeability analysis,pharmacokinetic and brain distribution study in mice of imperatorin,isoimperatorin and cnidilin in Radix Angelicae Dahuricae

Coumarins are important constituents of Radix Angelicae Dahuricae, a well-known traditional Chinese medicine possess several known bioactivities with potentials in the treatment of central nervous system diseases. By using an HPLC–MS/MS method, we analyzed the in vivo plasma and brain pharmacokinetics of three ingredients of coumarins, including imperatorin, isoimperatorin and cnidilin in mice after oral administration of Dahuricae extract at doses of 800 mg/kg. The biosamples were prepared using acetonitrile precipitation and the separation was achieved on an XDB-C18 column by gradient elution. The BBB permeability and P-gp-mediated efflux were further examined in Madin Canine kidney cells transfected with full length cDNA for human multidrug resistance gene1 (MDCKII-MDR1). Our results demonstrate that the method has excellent and satisfactory selectivity, sensitivity, linearity, precision, and accuracy for simultaneous determination of imperatorin, isoimperatorin and cnidilin. The pharmacokinetics parameters were determined by using noncompartmental analyses, including the AUC_(0 − t) in plasma (1695.22, 1326.45 and 636.98 mg*h/L), the AUC_(0 − t) in brain (1812.35, 2125.17 and 1145.83 ng*h/g) as well as the T_1/2 in plasma (0.66, 0.82, 0.97 h) and brain (0.96, 1.1, 0.99 h) for imperatorin, isoimperatorin and cnidilin, respectively, suggesting that the three coumarins could easily pass through the BBB in vivo. In the in vitro model we observed high permeability of imperatorin and isoimperatorin with the P-gp-mediated efflux ratios of 0.53 and 0.06, as well as medium permeability of cnidilin with 0.82. All data suggest that these three coumarins have high BBB permeability and have pharmacokinetic potentials for the treatment of central nervous system diseases.