Selenium increases antimony uptake in ramie (Boehmeria nivea L.) by enhancing the physiological, antioxidative, and ionomic mechanisms.

Ramie (Boehmeria nivea L.) is a promising phytoremediation candidate due to its high tolerance and enrichment capacity for antimony (Sb). However, challenges arise as Sb accumulated mainly in roots, complicating soil extraction. Under severe Sb contamination, the growth of ramie may be inhibited. Strategies are needed to enhance Sb accumulation in ramie's aboveground parts and improve tolerance to Sb stress. Considering the beneficial effects of selenium (Se) on plant growth and enhancing resistance to abiotic stresses, this study aimed to investigate the potential use of Se in enhancing Sb uptake by ramie. We investigated the effects of Se (0.5, 1, 2, 5, or 10 µM) on ramie growth, Sb uptake and speciation, antioxidant responses, and ionomic profiling in ramie under 10 mg/L of SbIII or antimonate (SbV) stresses. Results revealed that the addition of 0.5 µM Se significantly increased shoot biomass by 75.73% under SbIII stress but showed minimal effects on shoot and root length in both SbIII and SbV treatments. Under SbIII stress, 2 µM Se significantly enhanced Sb concentrations by 48.42% in roots and 62.88% in leaves. In the case of SbV exposure, 10 µM Se increased Sb content in roots by 42.57%, and 1 µM Se led to a 91.74% increase in leaves. The speciation analysis suggested that Se promoted the oxidation of SbIII to less toxic SbV to mitigate Sb toxicity. Additionally, Se addition effectively minimized the excess reactive oxygen species produced by Sb exposure, with the lowest malondialdehyde (MDA) content at 0.5 µM Se under SbIII and 2 µM Se under SbV, by activating antioxidant enzymes including superoxide dismutase, catalase, peroxidase, and glutathione peroxidase. Ionomic analysis revealed that Se helped in maintaining the homeostasis of certain nutrient elements, including magnesium, potassium (K), calcium (Ca), iron (Fe), and copper (Cu) in the SbIII-treated roots and K and manganese (Mg) in the SbV-treated roots. The results suggest that low concentrations of Se can be employed to enhance the phytoremediation of Sb-contaminated soils using ramie.

The influence of residual pectin composition and content on nanocellulose films from ramie fibers: Micro-nano structure and physical properties.

In this study, cellulose nanofibril (CNF) films from ramie fibers were prepared with different pectin compositions and contents, and the influence of residual pectin on the overall performances of CNF films was evaluated. There was no significant effect of the residual pectin composition on the properties of obtained CNF films. However, when the content of residual pectin was increased from 0.45 % to 9.16 %, the surface area and water absorption of CNF films were increased from 0.2223 to 0.3300 m2/g, and from 93.51 % to 122.42 %, respectively. Pectin covers the CNF surface and act as a physical barrier between the cellulose fibrils; thus the nanocellulose films with high pectin content will have a loose and porous structure, resulting in a high surface area and a high water absorption. Besides, with the residual pectin content decreasing from 9.16 % to 0.45 %, the UVA light transmittance and tensile strength of CNF films were increased from 30.6 % to 59.9 %, and from 37.67 to 100.26 MPa, respectively. After removal of amorphous pectins in CNFs, the low pectin containing CNFs are able to pack more compactly to form a strong and thin film. This paper provides guidance for the preparation of CNF films with different performance requirements.

Exogenous plant growth regulator and foliar fertilizers for phytoextraction of cadmium with Boehmeria nivea [L.] Gaudich from contaminated field soil.

As a enrichment plant, ramie can be used for the phytoremediation of cadmium (Cd)-contaminated soil. However, it is worth exploring the role of plant growth regulators and foliar fertilizers in the process of plant growth and development and Cd adsorption. By measuring the agronomic traits, Cd content of aboveground and underground ramie, calculating the Cd transfer coefficient (TF) and Cd bioconcentration factors (BCF), and the correlation between various indicators. This study examined the effects of plant growth regulators and foliar fertilizers on ramie's capacity for Cd accumulation and transportation. Plant growth regulators and foliar fertilizers increased the Cd content of the aboveground ramie, reduced the Cd content of the underground ramie, and increased the TF. Among them, GA-1 increased the Cd content of the aboveground ramie to 3 times more than that of the control and reduced the Cd content of the underground ramie by 54.76%. Salicylic acid (SA) increased the Cd content of the aboveground ramie to three times more than that of the control. The combination of GA and foliar fertilizer reduced the Cd content of the aboveground and underground ramie and the TF and BCF of the underground ramie. After the hormones were sprayed, the TF of ramie had a significant positive correlation with the Cd content of the aboveground ramie; the BCF of the aboveground ramie had a significant positive correlation with the Cd content and TF of the aboveground ramie. The results indicate that Brassinolide (BR), gibberellin (GA), ethephon (ETH), polyamines (PAs), and salicylic acid (SA) have different effects on the enrichment and transport of Cd in ramie. This study provided an effective method to improve the capacity for ramie to adsorb heavy metals during cultivation.

Elucidating structure of pectin in ramie fiber to customize enzyme cocktail for high-efficiency enzymatic degumming.

Pectin is one of the main components of bast fiber including ramie fiber, and must be removed before use. Enzymatic degumming is the preferred process as it is an environment-friendly, simple and controllable process for ramie degumming. However, an important problem limiting wide application of this process is the high cost due to the low efficiency of enzymatic degumming. In this study, pectin samples were extracted from raw ramie fiber and degummed ramie fiber, respectively, and their structures were characterized and compared to allow tailoring of an enzyme cocktail for degrading the pectin. It was elucidated that pectin from ramie fiber is composed of low esterified homogalacturonan (HG) and low branched rhamnogalacturonan I (RG-I), and the ratio of HG/RG-I is 1.72:1. Based on the pectin structure, potential enzymes to be used for enzymatic degumming of ramie fiber were proposed and an enzyme cocktail was customized. Degumming experiments confirmed that the customized enzyme cocktail can effectively remove pectin from ramie fiber. To our knowledge, this is the first time the structural characteristics of pectin in ramie fiber have been clarified, and it also provides an example of tailoring a specific enzyme system to achieve high-efficiency degumming for biomass containing pectin.

Ramie leaf Extract Alleviates Bone Loss in Ovariectomized Rats-The Involvement of ROS and Its Associated Signalings.

Ramie leaf (Boehmeria nivea L.) has been traditionally used to treat gynecological and bone-related disorders. This study aims to evaluate the effect of Ramie leaf extracts (RLE) against osteoporosis in ovariectomized (OVX) rats. Female SD rats aged seven weeks were randomly assigned into five OVX and a sham-operated (sham) group. OVX subgroups include OVX, vehicle-treated OVX group; E2, OVX with 100 µg/kg 17ß-estradiol; and RLE 0.25, 0.5, and 1, OVX rats treated with 0.25, 0.5, and 1 g/kg/day RLE, respectively. Two weeks into the bilateral ovariectomy, all the rats were orally administered with or without RLE daily for 12 weeks. OVX rats administered with RLE showed higher bone density, relatively low tartrate-resistant acid phosphatase (TRAP)-positive osteoclasts, and lower reactive oxygen species (ROS) within bone tissues compared to vehicle-treated OVX rats. Furthermore, supplementation of RLE improved bone mineral density (BMD) and bone microstructure in the total femur. RLE prevented RANKL-induced osteoclast differentiation and expression of osteoclastogenesis-related genes such as Cal-R, MMP-9, cathepsin K, and TRAP in RANKL-induced RAW264.7 cells. Moreover, RLE administration lowered the intracellular ROS levels by reducing NADPH oxidase 1 (NOX-1) and 4-hydroxynonenal (4HNE). These results suggest that RLE alleviates bone mass loss in the OVX rats by inhibiting osteoclastogenesis, where reduced ROS and its associated signalings were involved.

UPLC-QToF-MS based fingerprinting of polyphenolic metabolites in the bark extract of Boehmeria rugulosa Wedd.

Boehmeria rugulosa Wedd. is an evergreen tree of Urticaceae family. Its bark has been extensively used in ethno-medicinal system for various ailments such as bone fracture, sprain, snakebite, and wound healing. Phyto-metabolites, which are considered as the principle components for biological activities, have been least explored for this plant. The present work investigated metabolite profiling of the stem bark of B. rugulosa in water extract using Ultra Performance Liquid Chromatography Quadrupole Time of Flight Mass Spectrometry (UPLC-QToF-MS) technique coupled with the UNIFI platform. We identified, for the first time, 20 polyphenolic metabolites belonging to seven groups: caffeoylquinic acids, coumaroylquinic acids, flavan-3-ols, oligomeric flavonoids, caffeic acid derivatives, coumaric acid derivative, and flavone glycoside in the B. rugulosa extract. UNIFI informatics-coupled UPLC-QToF-MS platform aids in the quick identification and fragmentation pattern of metabolites, with higher degree of reproducibility. The present study provides a chemical and therapeutic basis for further exploration of B. rugulosa as a valuable source of phytochemicals that could be instrumental in deciphering its ethno-medicinal utility for various human diseases.

Beneficial role of Boehmeria nivea in health and phytochemical constituents.

The leaf and stem extracts of Boehmeria nivea (BN) collected from three different regions in Korea were screened for their antioxidant, neuroprotective, estrogenic, insulin secretion, and α-glucosidase inhibitory activity. We also examined whether BN extracts regulate cancer cell growth, inflammatory-related gene expression, and lipid accumulation in cellular system. Leaf extracts possessed greater antioxidant, anti-proliferative in cancer cells, neuroprotective, estrogenic activity, and inhibitory effect on pro-inflammatory gene expression than stem extracts. Leaf and stem extracts inhibited lipid accumulation in three T3-L1 adipocytes but did not affect glucose-stimulated insulin secretion in INS-1 cells. We isolated and identified the phytochemical constituents in the n-butanol and ethyl acetate fractions of BN leaves by combining silica gel column chromatography with mass spectrometry and 1 H- and 13 C-NMR analysis. The active compounds (caffeic acid, isoquercitrin, p-coumaric acid, and rutin) exhibited ABTS and DPPH radical scavenging activity, which may contribute to the biological activities of BN leaf extract. An analytical method was developed to quantify marker compounds for the discrimination of BN collected from different regions. Our results support the use of this analysis method for accurate identification and quantification of marker compounds in BN for the development of functional foods. PRACTICAL APPLICATIONS: Boehmeria nivea (BN) has been used as a raw material for the textile industry or traditional herbal medicine. The current study established the biological activities and active components of BN. Our results showed that BN leaf and stem extracts exhibit antioxidant, neuroprotective, and estrogenic activity. BN leaf extract also inhibited cancer cell growth, inflammatory mediators and cytokines production, and lipid accumulation in vitro. Moreover, the bioactive compounds, such as caffeic acid, isoquercitrin, p-coumaric acid, and rutin, exert ABTS and DPPH radical scavenging activities. Therefore, BN could potentially be a promising source of bioactive phytochemicals for the development of functional foods or drugs.

Effects of residual pectin composition and content on the properties of cellulose nanofibrils from ramie fibers.

In this study, a series of pretreatment methods (pectin extraction) were employed to prepare cellulose nanofibrils (CNFs) with different pectin composition and contents from ramie fibers. The comprehensive effects of residual pectin on the final properties of CNFs were investigated. The residual pectin did not have a significant effect on the size distribution of the obtained CNFs. While the presence of pectin led to higher zeta potential value, improved dispersion stability and enhanced storage modulus of the CNF dispersion, with the sodium carbonate extraction method showing the greatest impact. The possible mechanism for enhanced dispersion stability of CNFs is the formation of self-assembled hierarchical pectin-hemicellulose/lignin-cellulose nanostructures, offering abundant electrostatic repulsion and steric hindrance between the nanoparticles. This work provides guidelines for the tailored production of CNFs to meet the requirements for different applications.

Probing the transcriptome of Boehmeria nivea reveals candidate genes associated with the biosynthesis of chlorogenic acid.

Boehmeria nivea (L.) Gaudich is used in traditional Chinese medicine. Chlorogenic acids are major medically active components of Boehmeria nivea, which can be used clinically to treat hyperglycemia, pneumonia, and cancer. To identify the genes involved in chlorogenic acid biosynthesis, we analyzed transcriptome data from leaf, root, and stem tissues of Boehmeria nivea using the Illumina Hi-Seq 4000 platform. A total of 146,790 unigenes were obtained from Boehmeria nivea, of which 106,786 were annotated in public databases. In analyses of the KEGG (Kyoto Encyclopedia of Genes and Genome) database, 484 unigenes that encode the five key enzymes involved in chlorogenic acid biosynthesis were identified, and shikimate O-hydroxycinnamoyl transferase was spatially simulated. Some of these key enzyme unigenes expression levels were verified by RT-qPCR (real-time quantitative Polymerase Chain Reaction). Furthermore, multiple genes encoding plant resistance proteins or transcription factors were identified and analyzed. Differentially expressed genes were identified by performing pairwise comparison of genes between tissues. This study increases the number of public transcript datasets of this species and identifies candidate genes related to the biosynthesis of chlorogenic acid, laying a foundation for the further exploration of this pathway in Boehmeria nivea.

Phenanthroindolizine alkaloids from Boehmeria sieboldiana leaves exhibit cytotoxicity against human cancer cell lines.

To explore useful natural compounds from indigenous medicinal plants, the cytotoxic properties from a methanolic extract of Boehmeria sieboldiana leaves against human cancer cell lines were isolated in the present study. After purification of the extract, seco-dehydroantofine B (1) together with two known phenanthroindolizine alkaloids, seco-dehydroantofine A (2) and septicine (3), were isolated. The structure of seco-dehydroantofine B was elucidated by performing comprehensive one- and two-dimensional nuclear magnetic resonance spectroscopy and high-resolution electrospray ionization mass spectrometry. The cytotoxicity of these compounds against five human tumor cell lines was evaluated. Compound 3 exhibited anti-tumor activity at IC50 values of 50.0, 66.9, 50.0, and 153.7 µM against MKN1, SAS, HL-60, and THP-1 cells, respectively.

Structure of an Alkaline Pectate Lyase and Rational Engineering with Improved Thermo-Alkaline Stability for Efficient Ramie Degumming.

Alkaline pectate lyases have biotechnological applications in plant fiber processing, such as ramie degumming. Previously, we characterized an alkaline pectate lyase from Bacillus clausii S10, named BacPelA, which showed potential for enzymatic ramie degumming because of its high cleavage activity toward methylated pectins in alkaline conditions. However, BacPelA displayed poor thermo-alkaline stability. Here, we report the 1.78 Å resolution crystal structure of BacPelA in apo form. The enzyme has the characteristic right-handed ß-helix fold of members of the polysaccharide lyase 1 family and shows overall structural similarity to them, but it displays some differences in the details of the secondary structure and Ca2+-binding site. On the basis of the structure, 10 sites located in flexible regions and showing high B-factor and positive ΔTm values were selected for mutation, aiming to improve the thermo-alkaline stability of the enzyme. Following site-directed saturation mutagenesis and screening, mutants A238C, R150G, and R216H showed an increase in the T5015 value at pH 10.0 of 3.0 °C, 6.5 °C, and 7.0 °C, respectively, compared with the wild-type enzyme, interestingly accompanied by a 24.5%, 46.6%, and 61.9% increase in activity. The combined mutant R150G/R216H/A238C showed an 8.5 °C increase in the T5015 value at pH 10.0, and an 86.1% increase in the specific activity at 60 °C, with approximately doubled catalytic efficiency, compared with the wild-type enzyme. Moreover, this mutant retained 86.2% activity after incubation in ramie degumming conditions (4 h, 60 °C, pH 10.0), compared with only 3.4% for wild-type BacPelA. The combined mutant increased the weight loss of ramie fibers in degumming by 30.2% compared with wild-type BacPelA. This work provides a thermo-alkaline stable, highly active pectate lyase with great potential for application in the textile industry, and also illustrates an effective strategy for rational design and improvement of pectate lyases.

Physicochemical properties of a new starch from ramie (Boehmeria nivea) root.

A new starch was isolated from ramie root, and its physicochemical properties were investigated. Ramie dry root contained 45.9% starch. Starch had truncated, ellipsoidal, and spherical granule shapes with size from 7 to 30 µm and D[4,3] about 14.1 µm. Starch contained 38.9% apparent amylose content and 22.4% true amylose content, exhibited B-type crystallinity, and had 26.6% relative crystallinity, 0.82 ordered degree, and 9.2 nm lamellar thickness. Starch had 71.8 °C gelatinization peak temperature and 15.6 J/g gelatinization enthalpy, and exhibited 31.4 g/g swelling power and 17.1% water solubility at 95 °C. Starch had peak, hot, breakdown, final, and setback viscosities at 3048, 2768, 279, 4165, and 1397 mPa s, respectively, and showed peak time at 4.36 min and pasting temperature at 75.0 °C. The native, gelatinized, and retrograded starches contained 15.1%, 94.0%, and 86.5% rapidly digestible starch and 83.3%, 4.0%, and 10.7% resistant starch, respectively. Compared with potato and rice starches, ramie starch was somewhat similar to potato starch but significantly different from rice starch in starch component, crystalline structure, and functional properties. Therefore, ramie starch exhibited the potential to be used as a thickening agent, resistant-digesting food additive, and alternative to potato starch in food and nonfood industries.

Order in cellulosics: Historical review of crystal structure research on cellulose.

The structure determination of cellulose represents a significant part in the history of polymer science. On one side, cellulose as a natural and renewable material has valuable physical properties which can be improved with detailed knowledge of the structure. On the other side, it is produced by nature as fibers, e.g. ramie, cotton etc., most suitable for structural studies by X-ray diffraction after their discovery in 1912 and was regarded by scientists as predestined for such an undertaking. The progress utilizing appropriate methods and models will be pursued in this investigation from the beginning of the structural research in 1913 till the proposal of realistic structures of cellulose in the 1970s. Different basic units of the crystal structure are discussed. Many structural models for native cellulose have been suggested over times with parallel or antiparallel running chains in the fibers and supported or abandoned. In this historical review on order in cellulosics, the crystal structure research throughout the times will be critically evaluated and the improvements in experiments and methods discussed. This subject is not restricted to cellulose rather can be extended to further polymer fibers.

In Silico, In Vitro, and In Vivo Antitumor and Anti-Inflammatory Evaluation of a Standardized Alkaloid-Enriched Fraction Obtained from Boehmeria caudata Sw. Aerial Parts.

In the context of the cancer-inflammation relationship and the use of natural products as potential antitumor and anti-inflammatory agents, the alkaloid-enriched fraction of Boehmeriacaudata (BcAEF) aerial parts was evaluated. In vitro antiproliferative studies with human tumor cell lines showed high activity at low concentrations. Further investigation on NCI-H460 cells showed an irreversible effect on cell proliferation, with cell cycle arrest at G2/M phase and programmed cell death induction. Molecular docking studies of four alkaloids identified in BcAEF with colchicine's binding site on ß-tubulin were performed, suggesting (-)-C (15R)-hydroxycryptopleurine as the main inductor of the observed mitotic death. In vivo studies showed that BcAEF was able to reduce Ehrlich tumor volume progression by 30 to 40%. Checking myeloperoxidase activity, BcAEF reduced neutrophils migration towards the tumor. The in vivo anti-inflammatory activity was evaluated by chemically induced edema models. In croton oil-induced ear edema and carrageenan (CG)-induced paw edema models, BcAEF reduced edema around 70 to 80% together with inhibition of activation and/or migration of neutrophils to the inflammatory area. All together the results presented herein show BcAEF as a potent antitumor agent combining antiproliferative and anti-inflammatory properties, which could be further explored in (pre)clinical studies.

Extract of Boehmeria nivea Suppresses Mast Cell-Mediated Allergic Inflammation by Inhibiting Mitogen-Activated Protein Kinase and Nuclear Factor-κB.

Mast cells are effector cells that initiate allergic inflammatory immune responses by inducing inflammatory mediators. Boehmeria nivea (Linn.) Gaudich is a natural herb in the nettle family Urticaceae that possesses numerous pharmacological properties. Despite the various pharmacological benefits of Boehmeria nivea, its effects on allergic inflammation have not yet been determined. Here, we investigated the effect of the ethanol extract of Boehmeria nivea (BNE) on degranulation rat basophilic leukemia (RBL)-2H3 mast cells stimulated with anti-dinitrophenyl (anti-DNP) and bovine serum albumin (BSA) during immunoglobulin E (IgE)-mediated allergic immune response. The results showed inhibition of the release of ß-hexosaminidase and histamine from the cells. BNE suppressed pro-inflammatory cytokines (Tumor necrosis factor (TNF)-α, Interleukin (IL)-1ß, and IL-6) and reduced T helper (Th)2 cytokine IL-4 expression and/or secretion correlated with the downregulation of p38, extracellular signal-regulated kinases (ERK) mitogen-activated protein kinase (MAPK), and nuclear factor-κB (NF-κB) signaling pathways in treated RBL-2H3 mast cells. In passive cutaneous anaphylaxis, treatment with BNE during IgE-mediated local allergic reaction triggered a reduction in mouse ear pigmentation and thickness. Taken together, these results indicated that BNE suppressed mast cell-mediated inflammation, suggesting that BNE might be a candidate for the treatment of various allergic disorders.

Thermostable and alkalistable exopolygalacturonase of Bacillus pumilus dcsr1: Characteristics and applicability.

The bioactive form of thermostable and alkali stable pectinase of Bacillus pumilus dcsr1 is a homodimer of the molecular mass of 60 kDa with a pI of 4.6. The enzyme is optimally active at 50 °C and pH 10.5, and its Michaelis constant (Km), maximum rate of reaction (Vmax), activation energy (Ea), and temperature quotient (Q10) values (for citrus pectin) are 0.29 mg mL-1, 116 µmole mg-1 min-1, 74.73 KJmol-1 and 1.57, respectively. The enzyme has a shelf life of one and a half years at room temperature as well as 4 °C. The activity of the enzyme is stimulated by Mn2+ and Ca2+ and inhibited by Hg+, Cd2+, Co2+, Zn2+, Fe2+, Pb2+, EDTA and urea to a varied extent. The conformational studies of the enzyme revealed a high ß-sheet content in the bioactive dimer, and high α-helix in the inactive monomer. The Circular Dichroism (CD) spectra of the dimer in the presence of inhibitors suggested a marked decrease in ß-sheet, and a significant increase in α-helix, suggesting a key role of ß-sheets in the enzyme catalysis. Based on the end product analysis, the enzyme is an exopolygalacturonase with a unique ability of transglycosylation. When ramie fibers were treated with the enzyme, removal of gummy material (pectin) was visible, confirming its applicability in the degumming process.

Eco-friendly scouring of ramie fibers using crude xylano-pectinolytic enzymes for textile purpose.

This study was carried out to demonstrate the biotechnological potential of xylano-pectinolytic enzymes on scouring of ramie fibers. Optimization of bioscouring process showed a maximum effect of enzymes with 50-mM strength of buffer, pH 8.5, fibers to liquid ratio of 1 : 20 (g:ml). Xylanase and pectinase dosage of 7.5 and 3.0 IU, respectively, was found to be best for removal of xylan and pectin impurities, after treatment time of 1.5 h, at 50 °C temperature and 55 rpm agitation rate. EDTA and Tween 80 at concentration of 1.5 mM and 1.25 %, respectively, were found to be the best for effective removal of impurities, in order to improve hydrophilicity of the fibers. After bioscouring, brightness and whiteness values of bioscoured fibers were increased by 9.72 and 7.10% in comparison with control fibers. After enzymatic scouring, a reduction of 14.45 % in yellowness was also seen in ramie fibers. Enzymatic treatment resulted in 6.97% increased brightness, 10.64% increased whiteness, and 4.11% decreased yellowness as compared with scoured ramie fibers. The results indicated that scouring using xylanase and pectinase enzymes could be a substitute for chemical scouring technique. Enzymatic scouring is, therefore, environmentally sustainable and saves energy, also decreases the consumption of harmful chemicals used in alkaline scouring. This is the first report showing the effect of xylanase and pectinase enzymes, produced by a bacterial isolate, on physico-chemical and various optical properties of ramie fibers.

Assessment of ramie leaf (Boehmeria nivea L. gaud) as an animal feed supplement in P.R. China.

Tropical and subtropical regions were quite short of high-quality protein forage. Ramie (Boehmeria nivea L. Gaud) leaves as crop by products, are rich in protein and widely cultivated in tropical and subtropical regions. Thence, the development and utilization of ramie is of great significance to animal production in these regions. But it contained high level of tannins and total phenols, which may produce potentially adverse effect. It is very essential to evaluate the safety of ramie leaves before they are used as feed supplements. To evaluate potential toxic level of ramie leaf, control group, low dose and high dose (0, 1, 2 g/kg·BW) groups of ramie leaf were conducted orally in Sprague-Dawley rats (SD rats). Body weight, hematology, and histopathology was assessed during 28 d of treatment and 14 d of recovery period. The results showed that there were no toxic symptoms appeared in the treated and control groups. There were a few individual indicators showed abnormal, but most indices of body weight, organ weight ratios and hematology were normal. And compared to control group, it showed no significant differences (P>0.05). Histopathological examination of the high dose group and control group showed that there was no lesions related to ramie administration. The pathological changes appeared in the liver, and lungs of rats in individual rat of both groups were common and spontaneous, and had no significant differences (P>0.05). These results suggest that under this experimental condition, up to 2 g/kg·BW intragastric administration of ramie leaf did not produce adverse effect to SD rats. These findings would provide available information for ramie leaf to utilize as a feed supplement, particularly in P.R. China.

Hexafluoroisopropanol-salt aqueous two-phase system for extraction and purification of chlorogenic acid from ramie leaves.

Hexafluoroisopropanol (HFIP) was used as a phase-separation solvent to develop novel alcohol-salt aqueous two-phase systems (ATPSs) with various salts. Phase diagram and effective excluded volume (EEV) study proved that HFIP has much better phase-separation ability compared to traditional small molecule alcohols (ethanol, isopropanol and n-propanol). Then, the HFIP-NaCl ATPS was applied for the extraction and purification of chlorogenic acid (CGA) from ramie leaves. Under the optimum conditions (2 M NaCl solution with pH 3.0, the volume ratio of NaCl solution to HFIP at 6, vortex time 5 s and centrifugation time 7 min), the extraction efficiency of CGA in the salt-rich phase was 99.3%, meanwhile the HFIP-rich phase could extract a large amount of impurities. Furthermore, the CGA product with the purity of 91.0% was obtained from the salt-rich phase by semi-preparative liquid chromatography and salt removal, and its chemical structure was identified. Compared with other ATPSs, the HFIP-NaCl ATPS consumed much less organic solvent and salt, but acquired much higher extraction efficiency and obvious impurity-removal effect. Therefore, the HFIP-based alcohol-salt ATPSs are promising in the extraction and purification of CGA and other polar compounds as well.

Biochar facilitated the phytoremediation of cadmium contaminated sediments: Metal behavior, plant toxicity, and microbial activity.

Cadmium (Cd) contamination in river sediments becomes increasingly serious, and phytoremediation has been used to remediate Cd contaminated sediments, but the remediation efficiency needs to be improved. In this study, tea waste derived biochar (TB) was used to facilitate the phytoremediation of Cd contaminated sediments. Results showed that TB at 100, 500 and 1000 mg kg-1 increased Cd accumulation and translocation in ramie seedlings by changing Cd speciation in sediments and altering the subcellular distribution of Cd in plant cells. TB at low contents alleviated Cd induced toxicity in ramie seedlings by promoting plant growth and mitigating the oxidative stress. In addition, the activities of urease-, phosphatase-, and catalase-producing microbes in the Cd contaminated sediments were promoted by the application of TB. These findings demonstrated that biochar at low concentrations could improve the phytoremediation efficiency and mitigating Cd-induced toxicity to plants and microbes in Cd contaminated sediments. This study herein provides a novel technological application of waste biomass in controlling and mitigating risks of heavy metals.