Echinacoside promotes bone regeneration by increasing OPG/RANKL ratio in MC3T3-E1 cells

Echinacoside (ECH), isolated from Cistanche tubulosa (Schrenk) R. Wight stems, was subjected to in vitro experiments to investigate its bioactivities on proliferation, differentiation and mineralization of the osteoblastic cell line MC3T3-E1. MTT assay, the alkaline phosphatase (ALP) activity and calcium deposition were determined, and the secretion of collagen I (COL I), osteocalcin (OCN), osteoprotegerin (OPG) and receptor activator of nuclear factor-κB ligand (RANKL) were also assayed by enzyme-linked immunosorbent assay (ELISA). The results showed that ECH caused a significant increase in cell proliferation, ALP activity, COL I contents, OCN levels and an enhancement of mineralization in osteoblasts at the concentration range from 0.01 to 10 nmol·L^− 1 (p < 0.05), suggesting that ECH has a stimulatory effect on osteoblastic bone formation or has potential activity against osteoporosis. In addition, the ratio of OPG/RANKL also could be enhanced by ECH. These findings provide the potent evidence that ECH can promote bone regeneration in cultured osteoblastic MC3T3-E1 cells, which might be done by elevating the OPG/RANKL ratio, and potential evidence for echinacoside to be a promising drug or a lead compound in the development of disease-modifying drug to prevent osteoporosis.     

Bilberry extract protect restraint stress-induced liver damage through attenuating mitochondrial dysfunction

The aim of the present study is to investigate the protective effect of bilberry extract on liver damage in restraint stressed mice. A remarkable increase in alanine aminotransferase (ALT) and reactive oxygen species (ROS) levels was observed in stressed mice. Treatment with bilberry extract restored ALT and ROS to normal levels, and enhanced mitochondrial complex II activity that was lowered in restraint stressed mice. The mitochondrial electron transfer chain (ETC)-related gene expression was measured by RT-PCR, and a significant up-regulation of complex II mRNA was observed for SDHA, B, C and D mRNA in bilberry extract-treated group compared with that in stressed group. Bilberry extract administration also profoundly elevated the Na⁺-K⁺-ATPase activity and mitochondria membrane potential (?Ψ_m), which was reduced in the stressed group. Bilberry extract exhibited protective effect by scavenging free radicals and attenuating mitochondrial dysfunction in the liver of restraint stressed mice. It may be used as a promising therapeutic agent in preventing and delaying the life-related disease.     

Mutagenesis, heterogeneous gene expression, and purification and amino acid substitution analyses of plant peroxidase, PrxA3a

We introduced mutations into prxA3a, a peroxidase gene of hybrid aspen, Populus kitakamiensis, to substitute the amino acid residues at the surface of the protein, and analyzed substrate specificities. PrxA3a and mutated enzymes heterogeneously gene expressed in Saccharomyces cerevisiae were purified by Ni affinity chromatography, hydrolysis of sugar chain (Endoglycosidase H_f) and gel filtration. The substrate specificities were altered by substituted amino acid residues. PrxA3a F77Y A165W acquired the substrate specificity to m-chlorophenol. PrxA3a F77Y and PrxA3a F77YA165W could polymerize sinapyl alcohol. In addition, PrxA3a A165W, F77Y, and F77YA165W improved cytochrome c oxidizing activity. These substituted amino acid residues should function as a catalytic site outside of the heme pocket.     

Seed storage behavior of <Emphasis Type="Italic">Knema attenuata</Emphasis>, an endemic species of Western Ghats,India

We performed desiccation and storage trials to better under- stand storage behavior of Knema attenuata seeds. Mature seeds with moisture content (MC) of 31% exhibited 73% germination. During the period of desiccation (open lab condition) seeds with MC 23% showed 40% germination. After further drying to MC 21% germination was reduced to 16%. Complete loss in viability resulted when seed moisture was reduced to 18%. The seeds stored at -10°C, 0°C, 10°C and 28±2°C (open lab condition) lost their viability within 10 days. Seeds stored in sealed polythene bags and moist sand retained viability for more days than did seeds stored under all other storage conditions. Sensitivity of seeds to lower temperature and desiccation suggest that the storage be- havior of K. attenuata seeds is recalcitrant. Seeds stored in moist condi- tions can, at best, be stored for a period of two months.     

细胞色素C与植物细胞编程性死亡

目前普遍认为:细胞色素C(Cyt c)从植物线粒体内向胞浆的释放与植物细胞编程性死亡(PCD)关系密切。与动物PCD过程相似,植物线粒体内Cyt c的释放通过线粒体通透性转换孔(MPTP)或直接通过线粒体外膜的孔蛋白——电位依赖型阴离子通道进行。活性氧(ROS)诱导MPTP的形成,释放Cyt c,导致呼吸电子传递链阻断,ATP合成解偶联,产生ROS,ROS反过来再刺激Cyt c的释放;细胞浆内高水平的Ca2 会触发MPTP的形成以及Cytc的释放。作为植物PCD的早期事件,Cyt c的释放激活了特异性半胱氨酸蛋白酶类的信号级联放大途径,最后产生以核小体DNA长度为基数的DNA片段。在植物中,已经鉴定了几个具有与动物细胞凋亡蛋白酶活化因子-1相同序列的植物基因产物;Cyt c的释放对PCD的激活作为细胞凋亡的古老机制,从多细胞生物的单细胞祖先继承并进化而来。     

Schisandrin B protects against solar irradiation-induced oxidative stress in rat skin tissue

Schisandrin B (Sch B) and schisandrin C (Sch C), but not schisandrin A and dimethyl diphenyl bicarboxylate, protected rat skin tissue against solar irradiation-induced oxidative injury, as evidenced by a reversal of solar irradiation-induced changes in cellular reduced glutathione and α-tocopherol levels, as well as antioxidant enzyme activities and malondialdehyde production. The cytochrome P-450-mediated metabolism of Sch B or Sch C caused ROS production in rat skin microsomes. Taken together, Sch B or Sch C, by virtue of its pro-oxidant action and the subsequent eliciting of a glutathione antioxidant response, may prevent photo-aging of skin.     

Role of mitochondrial permeability transition in human hepatocellular carcinoma Hep-G2 cell death induced by rhein

Rhein, a compound found as a glucoside in the root of rhubarb, is currently a subject of interest for its antitumor properties. The apoptosis of tumor cell lines induced by rhein was observed, and the involvement of mitochondria was established; however, the role of mitochondrial permeability transition (MPT) remains unknown. Here we report that MPT plays an important role in the apoptosis of human hepatocellular carcinoma Hep-G2 cells induced by rhein. After adding rhein to the isolated hepatic mitochondria, swelling effects and the leakage of Ca^2 + were observed. These alterations were suppressed by cyclosporin A (CsA), an MPT inhibitor. Furthermore, in Hep-G2 cells, the decrease of ATP production, the loss of mitochondrial transmembrane potential (MTP), the release of cytochrome c (Cyto c), and the activation of caspase 3 were also observed. These toxic effects of rhein can also be attenuated by CsA as well. Moreover, TUNEL assay confirmed that in the presence of CsA, rhein-induced apoptosis was largely inhibited. These results suggest that MPT plays a critical role in the pathogenesis of Hep-G2 cell injury induced by rhein, and imply that MPT may contribute to the anti-cancer activity of rhein.     

Schisandrin B protects against solar irradiation-induced oxidative injury in BJ human fibroblasts

The effects of schisandrin B (Sch B) and its analogs on solar irradiation-induced oxidative injury were examined in BJ human fibroblasts. Sch B and schisandrin C (Sch C) increased cellular reduced glutathione (GSH) level and protected against solar irradiation-induced oxidative injury. The photoprotection was paralleled by decreases in the elastases-type protease activity and matrix-metalloproteinases-1 expression in solar-irradiated fibroblasts. The cytochrome P-450-mediated metabolism of Sch B or Sch C caused ROS production. The results suggest that by virtue of its pro-oxidant action and the subsequent glutathione antioxidant response, Sch B or Sch C may offer the prospect of preventing skin photo-aging.     

Phenolic glycosides from Curculigo orchioides Gaertn.

Five new chlorophenolic glucosides, curculigine E (1), curculigine F (2), curculigine G (3), curculigine H (5), curculigine I (6) and one new phenolic glycoside, orcinoside H (4), together with eight known phenolic glycosides (7–14) were isolated from the Curculigo orchioides Gaertn. Their structures were established by spectroscopic techniques (IR, UV, MS, 1D and 2D NMR). The isolated phenolic glycosides were evaluated for antiosteoporotic activity against MC3T3-E1 cell line using MTT assays. Compounds 1, 2, 3, and 5 showed moderate antiosteoporotic activity with the proliferation rate of 10.1–14.1%.     

密度对黑腹果蝇胚细胞系L-2/M delta 2-3 冻存效果的影响

使用双翅目昆虫黑腹果蝇( Drosophila melanogaster Fallen )胚细胞系L-2/M delta 2-3作为研究材料,制备10个密度试验组(0.81×10⁶~2.88×10⁷ 个·mL^-1,每组跨度大于2.00×10⁶ 个·mL^-1),分别在冻后第6、10、14个月对各组细胞冻后活力、回复时间长短、以及生长状况进行观察和比较,研究密度因素对细胞系L-2/M delta 2-3长期冻存效果的影响。结果表明:冻存密度对细胞系L-2/M delta 2-3冻后活力和状态恢复均有显著影响( P<0.05 )。细胞冻后活力下降速率随冻存密度增高而减慢,复苏后细胞状态恢复所需时间较短。将L-2/M delta 2-3的冻存密度提高至1.3×10⁷ 个·mL^-1以上有利于细胞活力的保持与冻后的恢复。     

Triterpenoid saponins from Clematis tangutica and their cardioprotective activities

Phytochemical investigation of the whole plants of Clematis tangutica led to the isolation of three new triterpenoid saponins (1–3), together with four known saponins (4–7). Their structures were determined by extensive spectral analysis and chemical evidences. Compounds 1–7 were evaluated for their cardioprotective activities in cardiomyocytes anoxia/reoxygenation (A/R) model. The results showed that those saponins exhibited cardioprotective effects by decreasing the levels of creatine kinase-MB (CK-MB) and lactate dehydrogenase (LDH).     

Cycloartane glycosides from Astragalus plumosus var. krugianus and evaluation of their antioxidant potential

The methanol extracts of Astragalus plumosus var. krugianus Chamb. & Matthews afforded sixteen cycloartane glycosides among which krugianoside A, was never reported before. All compounds were evaluated for their cytotoxic activity in human skin fibroblast WS1 cells. For compounds exhibiting no significant effect on WS1 viability, the antioxidant potential was examined. Compounds 1 and 8 prevented elevation of ROS induced by t-BOOH, suggesting the potential activity of these compounds to protect fibroblasts from oxidative stress.     

Seed viability as a function of moisture and temperature in the recalcitrant rainforest species Baccaurea ramiflora (Euphorbiaceae)

Context

Fragmentation is a common and increasingly serious problem in tropical rainforests through the world, resulting in changes in species composition and subsequent species extinctions.

Aim

The aim of this study was to test the hypothesis that changes in species composition can originate from the different selection pressures on pioneer and climax seeds as the habitat changes.

Methods

Baccaurea ramiflora seeds were placed on the ground in six quadrats in five different habitats in the field, while graded temperature and relative humidity treatments in the laboratory were used to mimic the changed microclimate in rainforest fragments. Changes in seed moisture and viability were monitored.

Results

B. ramiflora seeds are recalcitrant and highly sensitive to both high temperatures and dehydration, with a reduced lifespan when temperature increased or relative humidity decreased, and viability is lost when dehydrated to 0.198 g/g or warmed to?≥?35 °C. Germination was delayed in non-rainforest conditions compared with the rainforest understory, and only one-third of the seeds assigned to an open site germinated and emerged.

Conclusion

The results support our hypothesis that recalcitrant-seeded species have a high dependence on intact rainforests, and that fragmentation and disturbance will reduce their germination and establishment while favoring orthodox-seeded pioneers.     

Effect of salt on ROS homeostasis, lipid peroxidation and antioxidant mechanisms in Pinus pinaster suspension cells

? In the Pinus genus, information on the effectiveness of oxidative defence mechanisms during exposure to salt is lacking. The effect of salt stress imposition on ROS homeostasis was investigated using maritime pine (Pinus pinaster Ait.) suspension cells as a model system.

? Cells were maintained in MS-based medium, exposed to salt (50, 100 and 150 mM NaCl) and analysed for biomass production, evidencing a decreasing growth capacity. Use of 100 mM NaCl imposed severe salt stress without affecting cell viability, being chosen for subsequent studies on the ROS homeostasis of salt shock-treated suspension cells.

? Increased total ROS levels were evident on the second day of salt exposure, but a superoxide ion transient burst was immediately noticeable. Additionally, lipid peroxide formation seemed to correlate with superoxide ion breakdown. In-gel superoxide dismutase activity evidenced a FeSOD homodimer with strongly increasing activity between hours 12–48 of salt stress imposition. Subsequently, P. pinaster Fe-Sod1 and csApx1 genes were isolated from a cDNA library and expression was shown to increase within 12–24 h.

? Results show that severe salt treatment generates oxidative stress in P. pinaster cells despite the induction of antioxidant systems, and suggest a putative involvement of ROS in salt stress signalling.

     

Population genetic structure and ecological niche modelling of the leafhopper Hishimonus phycitis

Witches’ broom disease of lime, caused by ‘Candidatus Phytoplasma aurantifolia’, is responsible for major losses of Mexican lime trees in Southern Iran, Oman and the United Arab Emirates. The causative phytoplasma is transmitted by the leafhopper, Hishimonus phycitis. We combined ecological niche modelling with environmental and genetic data for six populations of H. phycitis from Iran and one from Oman. The mitochondrial cytochrome c oxidase I (COI) gene and nine microsatellite DNA markers were used for the genetic analyses. Although the Oman population had specific haplotypes, the COI sequences were highly conserved among all populations studied. In contrast, the microsatellite data divided the populations from Iran and Oman into two separate clades. An analysis of molecular variance indicated a high level of variation within populations. The Mantel test showed no correlation between genetic and geographical distances. Gene flow values were small between the populations from Iran and north of Oman but significantly higher among the Iranian populations supporting the differentiation between Iran and Oman. In addition, we found that patterns of genetic divergence within Iranian populations were associated strongly with divergence in terms of their ecological niches. Data on six climatic variables, including elevation, were used to create ecological niche models. Our results suggest that the genetic differentiation of H. phycitis may be attributable to climatic conditions and/or geographical barriers.     

Regioselective acylation of 3-O-angeloylingenol by Candida antarctica Lipase B

Acylation of 3-O-angeloylingenol (1) with vinyl acetate, vinyl decanoate and vinyl cinnamate, catalyzed by Candida antarctica Lipase B, was investigated. In each case, compound 1 was quantitatively and regioselectively acylated to afford a single product, 3-O-angeloyl-20-O-acetylingenol (1a), 3-O-angeloyl-20-O-decanoylingenol (1b) and 3-O-angeloyl-20-O-cinnamoylingenol (1c), respectively. The structures of the novel compounds 1b–1c were determined by MS and NMR, and product 1a by comparison of RP-HPLC and TLC with a standard. Compounds 1b–1c induced a bipolar morphology of MM96L melanoma cells at a similar concentration as compound 1, as well as having activity in inhibiting the growth of MM96L melanoma cells.     

Flavonol-rich fractions of yaupon holly leaves (Ilex vomitoria, Aquifoliaceae) induce microRNA-146a and have anti-inflammatory and chemopreventive effects in intestinal myofibroblast CCD-18Co cells

Polyphenolics extracted from yaupon holly (Ilex vomitoria, Aquifoliaceae) (YH) leaves were investigated in human colon cells for their chemopreventive and anti-inflammatory activities. An activity-guided fractionation allowed the selection of YH flavonol-rich fraction due to its preferential inhibition of HT-29 colon cancer viability over the normal CCD-18Co colon cells. Quercetin and kaempferol 3-rutinosides, main components identified in this fraction, protected CCD-18Co cells against reactive oxidative species (ROS) in part due to increased activity of antioxidant enzymes. In addition, up-regulation of microRNA-146a (miR-146a) known as a negative regulator of pro-inflammatory NF-κB activation was the underlying molecular mechanism that protected CCD-18Co from inflammation.     

Drying and soaking pretreatments affect germination in pedunculate oak

? Pedunculate oak (Quercus robur L.) acorns are recalcitrant so they are difficult to store. Therefore, deterioration in quality during storage may reduce seedling yield and quality. In an attempt to address this, the effects of drying and soaking treatments on acorn germination were assessed.

? After harvesting on two occasions in October, the moisture content (MC) of acorns were adjusted using the following treatments: (1) fresh state or control (46–48% MC); (2) soaked in fresh state (46–48%); (3) dried (40–42%) (standard method); and (4) dried and then soaked (46–48%). The treated acorns were allowed to germinate at 15 °C, both before and after storage at ?3 °C.

? Treatment effects were generally consistent for each harvest date. Compared to the untreated controls, soaking alone and drying followed by soaking significantly increased germination both before and after storage. Storage reduced germination, except for the dried and soaked acorns.

? The results confirmed that drying and soaking improved acorn germination, but this was not mediated through an effect on acorn MC. In a separate nursery experiment, acorns given this treatment resulted in higher seedling yields and better quality plants than those given the standard treatment (dried to 40–42% MC).

     

Investigation of changes in compressed moso bamboo (<Emphasis Type="Italic">Phyllostachys pubescens</Emphasis>) after hot-press molding

In this study, molding moso bamboo strips to a curved shape using hot-press molding operation was explored. Bamboo strips with different thickness and moisture content (MC) were subjected to press molding under 120–210 °C for different time. Changes in the chemical components of bamboo were analyzed by Fourier-transform infrared spectroscopy (FTIR). Effect of MC on thermal mechanical behavior of bamboo was investigated using dynamic mechanical analysis (DMA). Results showed that the influencing degree of four variables on compression and recovery ratios decreased as: temperature?>?time?>?thickness?>?MC. Compression ratio increased and recovery ratio decreased dramatically when pressing temperature exceeded 180 °C. FTIR analysis indicated that polysaccharide (especially hemicelluloses) underwent a progressive thermal degradation during compression at 180 and 210 °C for 40 min, whereas relative content of lignin increased. DMA results showed that bamboo samples with a higher MC had a lower storage modulus value, confirmed water had a plasticizing effect. The loss factor of bamboo with higher MC (12 and 16%) exhibited two major transitions centred around 100 °C (α₁) and 50 °C (α₂), respectively. The temperature of these α transitions kept almost unchanged as moisture level increased from 12 to 16%. These findings provide fundamental information for the future preparation of curved bamboo as profiled components in engineered products.     

Germination response of alder and birch seeds to applied gibberellic acid and priming treatments in combination with chilling

The effects of seed moisture content (MC), gibberellic acid (GA₃) concentration, chilling and priming pretreatments on the germination of common alder (Alnus glutinosa) and downey birch (Betula pubescens) seeds were examined. After treatment, the seeds were allowed to germinate for 42 days at 15 °C or 20 °C (dark)/30 °C (light). Treatment responses were similar at both temperatures and in both species. GA₃ treatment of seeds in fully imbibed (FI) state for 30 days, or at the lower, target moisture content (TMC) for 30–90 days, significantly improved germination, but longer treatment periods reduced it (FI seeds) or had no effect (TMC seeds). Priming for two days improved germination in the FI seeds, but more than 4 days reduced it. Priming for up to 14 days had little effect on the germination of the TMC seeds.