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In this paper, Acer truncatum seed meal was hydrolyzed by alkaline protease method. The enzymatic hydrolysis time, enzyme dosage, pH, enzymatic hydrolysis temperature and solid-liquid ratio were investigated, and the yield of enzymatic hydrolysis polypeptide was taken as the evaluation index. On the basis of the single-factor experiments and the Box-Behnken central combination experimental design, the preparation process of alkaline protease hydrolyzed polypeptide of Acer truncatum seed meal was optimized. The amino acid composition, water absorption, oil absorption, foaming, emulsifying and surface hydrophobicity of the enzymatic hydrolyzed peptides were characterized. The results showed that the optimal enzymatic hydrolysis process was as follows: enzymatic hydrolysis time 3.3 h, pH10, enzyme dosage 3%, enzymatic hydrolysis temperature 55 ℃. Under the optimum preparation conditions, the yield of peptides hydrolyzed by alkaline protease from Acer truncatum seed meal was 40.13%±0.15%. The analysis of amino acid composition showed that the amount of eight essential amino acids contained in the enzymatic hydrolysis polypeptide was as high as 20.3%, which was much higher than the amount of essential amino acids recommended by FAO. In addition, the oil absorption of enzymatic hydrolysis polypeptide (4.553 g/g) was higher than that of soybean protein (2.61 g/g), its surface hydrophobicity (1365.3) was similar to that of soybean 7S globulin, and its emulsifying property and emulsifying stability were slightly lower than that of soybean protein isolate. The alkaline protease hydrolyzed polypeptide of Acer truncatum seed meal obtained in this study had good functional characteristics, which also showed that it could be used as a potential functional component in food, providing data and theoretical support for the new application and development of Acer truncatum seed meal.

With the purpose of improving the water solubility and oral bioavailability, ursolic acid nanoparticles (UANs) were prepared by the emulsion solvent evaporation method, and the nanosuspension was freeze-dried into powder. The optimal conditions for preparing nanoparticles were screened out using single-factor experiment. Take advantage of the optimal conditions, UA nanoemulsion had mean particle size (MPS) of 69.7 ± 15.6 nm and polydispersity index value (PI) of 0.005. The MPS of UA nanosuspension was gained at 100.2 ± 12.1 nm (PI = 0.005), after the organic solvent was removed by rotary evaporator. Finally, UANs possessing an MPS of 157.5 ± 28.0 nm (PI = 0.005) and zeta potential of 20.33 ± 1.67 mV were obtained after freeze-dried. UANs were investigated using SEM, XRD, DSC, TGA and further explored their equilibrium solubility, dissolution rate, solvent residue analysis, cellular antioxidant activity and oral bioavailability. All the results above showed that UA in UANs was in the amorphous state. The result of solubility test figured that the equilibrium solubility of UANs was 13.48 times in simulated gastric fluid (SGF), 11.79 times in simulated intestinal fluid (SIF) and 23.99 times in deionized water than raw UA. Accordingly, the dissolution rate of UANs in SGF and SIF had an apparent enhancement. The oral bioavailability of UANs increased 2.68 times than raw UA. UANs improved antioxidant activity toward cells compared with raw UA, and EC50 of UANs reduced 37.5 times than raw UA. The residual contents of trichloromethane and ethanol were separated up to the mustard of the ICH limit for class III and class II solvents. The results above indicated that UANs possesses a value of application on enhancement oral bioavailability.

The aim of the experiment was to prepare licorice flavonoids nanoparticles (LFNs) by a liquid antisolvent re-crystallization (LAR) method to improve its oral bioavailability. The experimental data suggested that the saturation solubility of LFNs was 25 times of raw licorice flavonoids (LFs), and at 720 min, the dissolution rates of LFNs in artificial intestinal juice and artificial gastric juice were 9.36 times and 11.66 times of raw LFs, respectively. Furthermore, the antioxidant capacity of LFNs was also superior to raw LFs, and the oral bioavailability of echinatin and licochalcone A in the LFNs was 10.63 times and 6.54 times higher than that of raw LF. LFNs had good biosafety for SD rats at dosages up to 800 mg/kg. Overall, the results manifested that LFNs can be helpful to the enhancement of its oral bioavailability and have a promising application value in the field of food and health care.

Luteolin (LUT, 3′,4′,5,7-tetrahydroxyflavon) is a low water-soluble flavonoid with multiple biogicially activities. In this study, high water-soluble LUT nanoparticle was prepared by liquid anti-solvent precipitation and vacuum freeze-dry technology to enhance bioavailability and inhibit liver microsomal peroxidation in rats. When the concentration of LUT in the nanoparticles was 10.7 ± 0.3% (w/w), the size of LUT nanoparticles in water was successfully decreased to 127.5 ± 23.1 nm, and the amorphous state was reached. Meanwhile, the saturated solubility of LUT nanoparticles in deionized water (149.69 ± 12.6 µg) increased by 132.74 times. On the basis of the abovementioned results, LUT nanoparticles achieved excellent results in dissolution rates, lipid oxidation in vitro and bioavailability. Finally, results showed that the small particle size and amorphous state of LUT nanoparticles considerably improved their solubility in the solution medium, thereby enhancing their antioxidant ability and bioavailability.

The present work aimed to apply the liquid antisolvent precipitation (LAP) method for preparing the apigenin nanoparticles and thereby improving the solubility and bioavailability of apigenin. The different experimental parameters on particle size were optimized through central composite design (CCD) using the Design-Expert® software. Under the optimum conditions, the particle size of the apigenin nanosuspension was about 159.2 nm. In order to get apigenin nanoparticles, the freeze-drying method was selected and the mannitol was used as a cryoprotectant. Then the solid state properties of the apigenin nanoparticles were investigated using scanning electron microscopy (SEM), differential scanning calorimetry (DSC), thermo gravimetric (TG), and X-ray diffraction (XRD). The results obtained displayed that the apigenin nanoparticles exhibited near-spherical shape and could be transformed into an amorphous form. In addition, the dissolving test, the bioavailability in rats, and the antitumor activity were also studied. The experimental results showed that the solubility of the apigenin nanoparticles were about 29.61 times and 64.81 times of raw apigenin in artificial gastric juice and in artificial intestinal juice, respectively, and the apigenin nanoparticles showed higher dissolution rates compared to raw apigenin, and was about 6.08 times and 6.14 times than that of raw apigenin in artificial gastric juice and in artificial intestinal juice. The oral bioavailability of apigenin nanoparticles was about 4.96 times higher than that of the raw apigenin, but the apigenin nanoparticles had no toxic effect on the organs of rats. In addition, the apigenin nanoparticles had a higher inhibition to HepG2 cells by lower IC50 than that of raw apigenin.

A continuous process based on the combination of ultrasounds and/or microwaves pretreatments followed by enzymatic hydrolysis and simultaneous extraction (EHSE) has been proposed to recover genipin from Eucommia ulmoides bark. At first, in the pretreatment step, the mixture of 1.0 g dried bark powder and 10 mL deionized water were irradiated by microwave under 500 W for 10 min. Then, in hydrolysis step, the optimal conditions were as follows: 0.5 mg/mL of cellulase concentration, 4.0 pH of enzyme solution, 24 h of incubation time and 40 °C of incubation temperature. After incubation, 10 mL ethanol was added to extract genipin for 30 min by ultrasound. After EHSE treatment, the yield of genipin could reach 1.71 μmol/g. Moreover, scanning electron micrographs illustrated that severe structural disruption of plant was obtained by EHSE. The results indicated that the EHSE method provided a good alternative for the preparation of genipin from Eucommia ulmoides bark as well as other herbs.

In view of the poor water solubility and low oral bioavailability of mangiferin (MG), in this study, the supercritical antisolvent (SAS) technology was used to prepare mangiferin microparticles (MG MPs) with N,N-dimethylformamide (DMF) as solvent and carbon dioxide as antisolvent, so as to improve its water solubility, antioxidant capacity and oral bioavailability. Four factors affecting the solubility of the MG MPs were investigated by orthogonal design (OAD), including precipitation pressure, precipitation temperature, MG concentration and feeding speed, and the optimal preparation conditions were determined by range and variance analysis (ANOVA). Under the optimal conditions, the spherical MG MPs with an average diameter of 532.8 nm was obtained, and the yield of the powder was about 95.3%. Scanning electron microscopy (SEM), fourier transform infrared spectroscopy (FTIR), X-Ray Diffractometry (XRD), differential scanning calorimetry (DSC), and thermal gravimetric (TG) were used to analyze the characteristics of the MG MPs. The results obtained showed that the chemical structure of the MG did not change before and after supercritical crystallization, but its particle size and crystallinity decreased significantly. The MG MPs had a higher solubility, and was about 4.26, 2.1 and 2.5 times than that of free MG in water, artificial gastric juice (AGJ) and artificial intestinal juice (AIJ), respectively. The dissolution rate of the MG MPs were also obviously higher than that of free MG. Furthermore, the bioavailability of the MG MPs in vivo was about 2.07 times higher than that of the free MG, and its antioxidant capacity was also much higher than that of free MG, which was close to vitamin C.

Sustainable development of renewable resources is a major challenge globally. Biomass is an important renewable energy source and an alternative to fossil fuels. Pyrolysis of biomass is a promising method for simultaneous production of biochar, bio-oil, pyroligneous acid (PA), and gaseous fuels. The purpose of this study was to investigate the pyrolysis process and products yields of Schisandra chinensis fruits with different pyrolysis powers. The obtained PA was extracted with organic solvents, including ethyl formate, dichloromethane, methanol and tetrahydrofuran. The antioxidant activities, including the free radical scavenging activity and ferric reducing power, of the PA extracts were investigated. The synthetic antioxidants butylated hydroxyanisole and butylated hydroxytoluene were used as positive controls. A dichloromethane extract of PA showed excellent antioxidant properties compared to the other extracts. The chemical compositions of the PA extracts were determined by GC-MS, and further proved that the dichloromethane extract had the best antioxidant characteristics among the extracts tested.

An ionic liquid-based microwave-assisted extraction method (ILMAE) was successfully applied for the extraction of taxifolin from Larix gmelinii. Different kinds of 1-alkyl-3-methylimidazolium ionic liquids with different kinds of cations and anions were studied and 1-butyl-3-methylimidazolium bromide was chosen as the optimal solvent for taxifolin extraction. The optimal conditions of ILMAE were determined by single factor experiments and Box-Behnken design as follows: [C4mim]Br concentration of 1.00 M, soaking time of 2 h, liquid-solid ratio of 15:1 mL/g, microwave irradiation power of 406 W, microwave irradiation time of 14 min. No degradation of taxifolin had been observed under the optimum conditions as evidenced from the stability studies performed with standard taxifolin. Compared with traditional solvent and methods, ILMAE provided higher extraction yield, lower energy and time consumption. The distribution of taxifolin in different parts of larch and the influences of age, orientation, and season on the accumulation of taxifolin were analyzed for the sufficient utilization of L. gmelinii.

An effective ionic liquid vacuum microwave-assisted method was developed for extraction of the thermo- and oxygen-sensitive glycosides salicin, hyperin and rutin from Populus bark due to the strong solvating effects of ionic liquids on plant cell walls. In this study, [C4mim]BF4 solution was selected as the extracting solution for extraction of the target analytes. After optimization by single factor experiments and response surface methodology, the optimum condition parameters were achieved, which included 1.0 M [C4mim]BF4, 2 h soaking time, −0.08 MPa vacuum, 20 min microwave irradiation time, 400 W microwave irradiation power and 25 mL/g liquid/solid ratio. Under the optimum conditions, higher extraction yields of salicin (35.53 mg/g), hyperin (1.32 mg/g) and rutin (2.40 mg/g) were obtained. Compared with other extraction methods, the developed method provided higher yields of the three target components after a relatively shorter extraction time (20 min). No obvious degradation of the target analytes was observed under the optimum conditions in performed stability studies and the proposed method had a high reproducibility. Meanwhile, after adsorption and desorption on macroporous D101 resin, the target analytes can be effectively separated from the [C4mim]BF4 ionic liquid extraction solution and the yields of salicin, hyperin and rutin were 89%, 82% and 84%, respectively. The recovered [C4mim]BF4 ionic liquid presented a good extraction effect on the three analytes after recycling five times.

In this study, epigallocatechin gallate (EGCG) palmitate was synthesized and its anti-porcine reproductive and respiratory syndrome virus (PRRSV) activity was studied. Specifically, EGCG palmitate was evaluated for its ability to inhibit PRRSV infection in MARC-145 cells when administered as pre-, post-, or co-treatment. EGCG and ribavirin were used as controls. The results showed that a 50% cytotoxic concentration (CC50) of EGCG, EGCG palmitate, and ribavirin was achieved at 2,359.71, 431.42, and 94.06 μM, respectively. All three drugs inhibited PRRSV in a dose-dependent manner regardless of the treatment protocol. EGCG palmitate exhibited higher cytotoxicity than EGCG, but lower cytotoxicity than ribavirin. EGCG palmitate anti-PRRSV activity was significantly higher than that of EGCG and ribavirin, both as pre-treatment and post-treatment. Under the former conditions and a tissue culture infectious dose of 10 and 100, the selectivity index (SI) of EGCG palmitate in the inhibition of PRRSV was 3.8 and 2.9 times higher than that of ribavirin when administered as a pre-treatment, while the SI of EGCG palmitate in the inhibition of PRRSV was 3.0 and 1.9 times higher than ribavirin when administered as a post-treatment. Therefore, EGCG palmitate is potentially effective as an anti-PRRSV agent and thus of interest to the pharmaceutical industry.

The aim of this study was to prepare 10-hydroxycamptothecin nanocolloidal particles (HCPTNPs) to increase the solubility of drugs, reduce the toxicity, improve the stability of the drug, and so forth. HCPTNPs was prepared by antisolvent precipitation (AP) method combined with high pressure homogenization (HPH), followed by lyophilization. The main parameters during antisolvent process including volume ratio of dimethyl sulfoxide (DMSO) and H2O and dripping speed were optimized and their effects on mean particle size (MPS) and yield of HCPT primary particles were investigated. In the high pressure homogeneous procedure, types of surfactants, amount of surfactants, and homogenization pressure (HP) were optimized and their influences on MPS, zeta potential (ZP), and morphology were analyzed. The optimum conditions of HCPTNPs were as follows: 0.2 mg/mL HCPT aqueous suspension, 1% of ASS, 1000 bar of HP, and 20 passes. Finally, the HCPTNPs via lyophilization using glucose as lyoprotectant under optimum conditions had an MPS of 179.6 nm and a ZP of 28.79 ± 1.97 mV. The short-term stability of HCPTNPs indicated that the MPS changed in a small range.

An efficient homogenate extraction technique was employed for extracting shikonin from Arnebia euchroma. The homogenate extraction procedure was optimized and compared with other conventional extraction techniques. The proposed method gave the best result with the highest extraction efficiency in the shortest extraction time. Based on single-factor experiments, a three-factor-three-level experimental design has been developed by Box-Behnken design. The optimal conditions were 78% ethanol as solvent, homogenate extraction time of 4.2 min, 10.3 liquid to solid ratio and two extraction cycles. Moreover, the proposed method was validated by stability, repeatability and recovery experiments. The developed homogenate extraction method provided a good alternative for the extraction of shikonin from A. euchroma. The results indicated that the proposed homogenate extraction was a convenient, rapid and efficient sample preparation technique and was environmental friendly. Furthermore, homogenate extraction has superiority in the extraction of thermally sensitive compounds from plant matrices.

In the present study, an improved method termed negative-pressure cavitation extraction (NPCE) followed by reverse phase high-performance liquid chromatography (RP-HPLC) was developed for the extraction and quantification of vindoline (VDL), catharanthine (CTR), vincristine (VCR) and vinblastine (VLB) from Catharanthus roseus leaves. The optimized method employed 60-mesh particles, 80% ethanol, a negative pressure of −0.075 MPa, a solid to liquid ratio of 1:20, 30 min of extraction and three extraction cycles. Under these optimized conditions, the extraction yields of VDL, CTR, VCR and VLB are 0.5783, 0.2843, 0.018 and 0.126 mg/g DW, respectively. These extraction yields are equivalent to those from the well-known ultrasonic extraction method and higher than the yields from maceration extraction and heating reflux extraction. Our results suggest that NPCE-RP-HPLC represents an excellent alternative for the extraction and quantification of vinca alkaloids for pilot- and industrial-scale applications.

The aim of this study was to investigate the antioxidant and free-radical scavenging activity of extract and fractions from various parts of Elsholtzia ciliata. The inflorescences, leaves, stems and roots of E. ciliata were extracted separately and two phenolic component enrichment methods: ethyl acetate-water liquid-liquid extraction and macroporous resin adsorption-desorption, were adopted in this study. The antioxidant activities of water extracts and fractions of E. ciliata were examined using different assay model systems in vitro. The fraction root E (purified by HPD300 macroporous resin) exhibited the highest total phenolics content (497.2 ± 24.9 mg GAE/g), accompanied with the highest antioxidant activity against various antioxidant systems in vitro compared to other fractions. On the basis of the results obtained, E. ciliata extracts can be used potentially as a ready accessible and valuable bioactive source of natural antioxidants.

In this study, Rosmarinus officinalis L. essential oil and three of its main components 1,8-cineole (27.23%), α-pinene (19.43%) and β-pinene (6.71%) were evaluated for their in vitro antibacterial activities and toxicology properties. R. officinalis L. essential oil possessed similar antibacterial activities to α-pinene, and a little bit better than β-pinene, while 1,8-cineole possessed the lowest antibacterial activities. R. officinalis L. essential oil exhibited the strongest cytotoxicity towards three human cancer cells. Its inhibition concentration 50% (IC50) values on SK-OV-3, HO-8910 and Bel-7402 were 0.025‰, 0.076‰ and 0.13‰ (v/v), respectively. The cytotoxicity of all the test samples on SK-OV-3 was significantly stronger than on HO-8910 and Bel-7402. In general, R. officinalis L. essential oil showed greater activity than its components in both antibacterial and anticancer test systems, and the activities were mostly related to their concentrations.

Camptothecin-20(s)-O-[N-(3’α,12’α-dihydroxy-24’-carbonyl-5’β-cholan)]-lysine (B2) is a novel camptothecin analogue. Our previous study had shown that it displayed higher cytoxicity activity towards hepatocellular carcinoma SMMC-7721 cells than camptothecin (CPT) in vitro. In this paper, the underlying mechanism of anti-proliferation of B2 towards SMMC-7721 cells was further examined. Cell growth inhibition of B2 was determined using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay; morphological changes were observed under Laser Scanning Confocal Microscope (LSCM); cell cycle distribution, apoptotic population, changes in mitochondrial membrane potential, intracellular calcium concentration and reactive oxygen species (ROS) production were determined by flow cytometry (FCM). Activities of caspase-3 and caspase-9 were measured, and the expression level of Bcl-2 and Bax proteins were analyzed by Western blot. The results suggested that B2 inhibited SMMC-7721 cell growth by causing cell cycle arrest at the S and G2/M phases, and induced apoptosis involving a mitochondrial pathway. B2 appears to cause a high induction of apoptosis on SMMC-7721 cells in vitro, which suggests it might be a potential drug for cancer therapy.

Typhonium giganteum Engl. (BaiFuzi) is one of the herbs commonly used in traditional Chinese medicine against cancer. In our previous studies, 37 compounds were identified the SFE-CO2 (supercritical fluid extraction with CO2) extract by GC-MS, including the four major components [β-sitosterol (40.22%), campesterol (18.45%), n-hexadecanoic acid (9.52%) and (Z,Z)-9,12-octadecadienoic acid (8.15%)]. The anti-cancer mechanisms of the SFE-CO2 extract from T. giganteum Engl. tubers have not been reported as yet. In this paper, the molecular mechanisms of the SFE-CO2 extract-mediated apoptosis in SMMC-7721 cells were further examined. SFE-CO2 extract inhibited the growth of SMMC-7721 cells in a time- and dose-dependent manner, arrested the cell cycle in the S phase and G2/M phase, and induced apoptosis. In addition, reactive oxygen species (ROS) increase, reduction of mitochondrial membrane potential, a rise in intracellular calcium levels were found in SMMC-7721 cells after treated with the extract. Western blot analysis showed that the extract caused down-regulation of Bcl-2 expression, and up-regulation of Bax expression. Moreover, caspase-3 and caspase-9 protease activity significantly increased in a dose-dependent manner. Collectively, our results showed that the SFE-CO2 extract from T. giganteum Engl. tubers induces apoptosis in SMMC-7721 cells involving a ROS-mediated mitochondrial signalling pathway.

A clean, practical and environmentally friendly synthesis in a homogeneous system has been developed for α-oxoketene S,S-acetals. A mixture of [Bmim]Cl and water was used as medium. The best economical and practical molar ratio of [Bmim]Cl to substrate was 4 to 1. With various types of 1,3-dicarbonyl compounds as substrate, the corresponding α-oxoketene S,S-acetals have been prepared under mild reaction conditions with yields of 53–74% after purification with silica gel column. [Bmim]Cl/water can be recycled several times.

In the present study, anti-IBV (infectious bronchitis virus) activities of (-)-pinenes were studied by MTT assay, as well as docking and molecular dynamic (MD) simulations. The CC50 values of (-)-α-pinene and (-)-β-pinene were above 10 mM. And the maximum noncytotoxic concentrations (TD0) of (-)-α-pinene and (-)-β-pinene were determined as 7.88 ± 0.06 and 6.09 ± 0.31 mM, respectively. The two compounds were found to inhibit IBV with an IC50 of 0.98 ± 0.25 and 1.32 ± 0.11 mM. The MTT assay showed that the inhibitions of (-)-pinenes against IBV appear to occur moderately before entering the cell but are much stronger occur after penetration of the virus into the cell. Molecular simulations indicated that (-)-α-pinene and (-)-β-pinene specifically interact with the active site which is located at the N terminus of phosphorylated nucleocapsid (N) protein, with the former being more potent than the latter. The binding energies of them are −36.83 and −35.59 kcal mol−1, respectively. Results presented here may suggest that (-)-α-pinene and (-)-β-pinene possess anti-IBV properties, and therefore are a potential source of anti-IBV ingredients for the pharmaceutical industry.

An efficient synthesis of a valuable intermediate of coenzyme Q10 by microwave-assisted Lewis acidic ionic liquid (IL)-catalyzed Friedel-Crafts alkylation is reported. The acidity of six [Etpy]BF4-based ionic liquids was characterized by means of the FT-IR technique using acetonitrile as a molecular probe. The catalytic activities of these ionic liquids were correlated with their Lewis acidity. With increasing Lewis acid strength of the ionic liquids, their catalytic activity in the Friedel-Crafts reaction increased, except for [Etpy]BF4-AlCl3. The effects of the reaction system, the molar fraction of Lewis acid in the Lewis acid ILs and heating techniques were also investigated. Among the six Lewis acid ionic liquids tested [Etpy]BF4-ZnCl2 showed the best catalytic activity, with a yield of 89% after a very short reaction time (150 seconds). This procedure has the advantages of higher efficiency, better reusability of ILs, energy conservation and eco-friendliness. The method has practical value for preparation of CoQ10 on an industrial scale.

Dongmei Zhao, Xiuhua Zhao, Yuangang Zu, Jialei Li, Yu Zhang, Ru Jiang, Zhonghua ZhangKey Laboratory of Forest Plant Ecology, Northeast Forestry University, Ministry of Education, Harbin, Heilongjiang, ChinaAbstract: Paclitaxel (Taxol®) is an important anticancer drug in clinical use for treatment of a variety of cancers. Because of its low solubility, it is formulated in high concentration in Cremophor EL® which induces hypersensitivity reactions. In this study, targeted delivery of paclitaxel-loaded nanoparticles was prepared by a desolvation procedure, crosslinked on the wall material of bovine serum albumin, and subsequently decorated by folic acid. The characteristics of the nanoparticles, such as amount of folate conjugation, surface morphology, drug entrapment efficiency, drug loading efficiency, and release kinetics were investigated in vitro. The targeting effect was investigated in vitro by cancer cell uptake of fluorescein isothiocyanate-labeled nanoparticles. The spherical nanoparticles obtained were negatively charged with a zeta potential of about -30 mV, and characterized around 210 nm with a narrow size distribution. Drug entrapment efficiency and drug loading efficiency were approximately 95.3% and 27.2%, respectively. The amount of folate conjugation was 9.22 µg/mg of bovine serum albumin. The folate-decorated nanoparticles targeted a human prostate cancer cell line effectively.Keywords: paclitaxel, bovine serum albumin, folate, nanoparticles, target delivery

Ten essential oils, namely, mint (Mentha spicata L.,Lamiaceae), ginger (Zingiber officinaleRosc.,Zingiberaceae), lemon (Citrus limon Burm.f.,Rutaceae), grapefruit (Citrus paradisi Macf., Rutaceae), jasmine (Jasminum grandiflora L.,Oleaceae), lavender (Mill.,Lamiaceae), chamomile (Matricaria chamomilla L., Compositae), thyme (Thymus vulgaris L., Lamiaceae), rose (Rosa damascena Mill.,Rosaceae) and cinnamon (Cinnamomum zeylanicumN. Lauraceae) were tested for their antibacterial activities towards Propionibacterium acnes and in vitro toxicology against three human cancer cell lines. Thyme, cinnamon and rose essential oils exhibited the best antibacterial activities towards P. acnes, with inhibition diameters of 40 ± 1.2 mm, 33.5 ± 1.5 mm and 16.5 ± 0.7 mm, and minimal inhibitory concentrations of 0.016% (v/v), 0.016% (v/v) and 0.031% (v/v), respectively. Time-kill dynamic procedures showed that thyme, cinnamon, rose, and lavender essential oils exhibited the strongest bactericidal activities at a concentration of 0.25% (v/v), and P. acnes was completely killed after 5 min. The thyme essential oil exhibited the strongest cytotoxicity towards three human cancer cells. Its inhibition concentration 50% (IC50) values on PC-3, A549 and MCF-7 tumor cell lines were 0.010% (v/v), 0.011% (v/v) and 0.030% (v/v), respectively. The cytotoxicity of 10 essential oils on human prostate carcinoma cell (PC-3) was significantly stronger than on human lung carcinoma (A549) and human breast cancer (MCF-7) cell lines.

Yuangang Zu, Yu Zhang, Xiuhua Zhao, Qi Zhang, Yang Liu, Ru JiangKey Laboratory of Forest Plant Ecology, Northeast Forestry University, Ministry of Education, Harbin, Heilongjiang, ChinaAbstract: Response surface methodology (RSM) was used to optimize the process of preparing bovine serum albumin (BSA) nanoparticles by desolvation, then the resulting BSA nanoparticles (BSANPs) were conjugated with folate to produce a drug carrier system that can specifically target tumors. The anticancer drug, vinblastine sulfate (VBLS), was loaded to this tumor-specific drug carrier system for the purpose of overcoming the nonspecific targeting characteristics and side effects of the drug. A central composite design was applied for modeling the process, which was composed of four independent variables, namely BSA concentration, the rate of adding ethanol (ethanol rate), ethanol amount, and the degree of crosslinking. The mean particle size and residual amino groups of the BSANPs were chosen as response variables. The interactive effects of the four independent variables on the response variables were studied. The characteristics of the nanoparticles; such as amount of folate conjugation, drug entrapment efficiency, drug-loading efficiency, surface morphology and release kinetics in vitro were investigated. Optimum conditions for preparing desired BSANPs, with a mean particle size of 156.6 nm and residual amino groups of 668.973 nM/mg, were obtained. The resulting folate-conjugated BSANPs (FA-BSANPs) showed a drug entrapment efficiency of 84.83% and drug-loading efficiency of 42.37%, respectively, and the amount of folate conjugation was 383.996 µM/g BSANPs. The results of this study indicate that using FA-BSANPs as a drug carrier system could be effective in targeting VBLS-sensitive tumors in the future.Keywords: bovine serum albumin, vinblastine sulfate, folate, targeted drug delivery, nanoparticles, response surface methodology

The cytotoxic activity of curcumin towards CCRF-CEM human T-cell leukemia cells was measured by the MTT assay. Tumor cells were more sensitive to the cytotoxic activity of curcumin or curcumin-Cu (II)compared to normal cells, and the IC50 ofcurcumin towards CCRF-CEM cells was 8.68 µM, and that ofcurcumin-Cu (II) was 8.14 µM. The cell cycle distribution of curcumin-treated CCRF-CEM cells was analyzed by flow cytometry. DNA damage induced by oxidants such as curcumin-Cu (II) ions is considered as one of the main causes of cell inactivation. Therefore, we analyzed the effect of curcumin on DNA damage by agarose gel electrophoresis and atomic force microscopy (AFM). Gel electrophoresis analyses showed that curcumin or Cu (II) alone failed to cause DNA damage in pBR322 plasmid DNA as compared with the normal plasmid. However, DNA plasmids were mostly damaged after treatment with curcumin of different concentrations in the presence of Cu (II). Two forms were observed by means of AFM: closed circular plasmids and linear plasmids. DNA damage induced by a combination of curcumin and Cu (II) was also found by agarose gel electrophoresis, which was applied as control method to verify the results obtained by AFM.

In order to hydrolyze geniposide and extract geniposidic acid, a novel microwave irradiation heating method for simultaneous hydrolysis and extraction for preparation of geniposidic acid from Eucommia ulmoides bark using basic imidazolide ionic liquid was developed in this work. Basic ionic liquid paired by imidazolide anion and 1-hexyl-3-methylimidazolium cation ([C6mim]Im) was used as a dual hydrolysis catalyst and extraction solvent in the proposed approach. The optimal concentration of [C6mim]Im was 0.4 mol/L. And the independent parameters optimized by Box-Behnken design (BBD) based on single-factor tests were as follows: microwave irradiation time of 20 min at power of 415 W and liquid-solid ratio of 15 mL/g. Yield of geniposidic acid was increased to 3.41±0.17 μmol/g. DM130 macroporous resin, after reaction, was selected to separate geniposidic acid from hydrolysate. The plant materials with different treatment were analyzed by scanning electron microscopy (SEM) and thermogravimetric analysis (TG). The analysis of SEM and TG further indicated that IMSHE is an efficient preparation method for geniposidic acid. It is reasonable to assume that the method and the dual hydrolysis catalyst and extraction solvent would offer an alternative for the preparation of some other kinds of bioactive substances from plant materials.

Ultrafine resveratrol (u-Res) particles were prepared through the SAS process. The orthogonal method was used to optimize the factors of the SAS process. The size of u-Res reached 0.68 μm under the optimum conditions. The characterization of the u-Res particles was tested by many analysis methods. The chemical structure of Res was unaffected by the SAS process. The degree of crystallinity of the u-Res particles greatly reduced. The purity of the u-Res particles increased from 98.5% to 99.2% during the SAS process. The u-Res particles had greater saturation solubility and dissolution rate than the raw-Res (r-Res) particles. The radical scavenging activity and bioavailability of the u-Res in vivo were 1.9 times of the r-Res.

Vitexin, a natural flavonoid found in many medicinal plants, is well known for its rich pharmacological activities. However, the poor water solubility of vitexin has limited its therapeutic application. The aim of this study was to prepare the nanoparticles of vitexin by combining the antisolvent precipitation (ASP) and high pressure homogenization (HPH) approaches followed by lyophilization for improving the dissolution rate of this poorly water-soluble drug. The effects of main factors influencing the mean particle size (MPS) of vitexin were investigated and optimized. Under optimum conditions, vitexin nanosuspensions with an MPS of 80.5 nm were obtained and then lyophilized to form nanoparticles. The obtained vitexin nanoparticles were further characterized by scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), mass spectrometry (MS), X-ray powder diffraction (XRPD), gas chromatography (GC) and dissolution testing. The results showed that the nanoparticles of vitexin were converted into an amorphous form, with its chemical structure unchanged. Additionally, the residual dimethyl sulfoxide (DMSO) is lower than the International Conference on Harmonization (ICH) limit for class 3 solvents. The dissolution rate of processed vitexin was significantly higher (5.58-fold) than that of raw drug. Overall, the combinative process we developed is an effective way to produce vitexin nanoparticles with markedly enhanced dissolution rate.

An ultrasound-assisted extraction (UAE) method using ethanol was applied for extracting arabinogalactan (AG) and dihydroquercetin (DHQ) simultaneously from larch wood, as a pretreatment for pulping and papermaking. The extraction parameters were optimized by a Box-Behnken experimental design with the yields of AG and DHQ as the response values. Under optimum conditions (three extractions, each using 40% ethanol, for 50 min, 200 W ultrasound power and 1∶18 solid-liquid ratio), the yields of AG and DHQ were 183.4 and 36.76 mg/g, respectively. After UAE pretreated, the wood chips were used for Kraft pulping (KP) and high boiling solvent pulping (HBSP). The pulping yield after pretreatment was higher than that of untreated (the pulping yields of untreated HBSP and KP were 42.37% and 39.60%, and the pulping yields of HBSP and KP after UAE-pretreated were 44.23% and 41.50% respectively), as indicated by a lower kappa number (77.91 and 27.30 for untreated HBSP and KP; 77.01 and 26.83 for UAE-pretreated HBSP and KP). Furthermore, the characteristics of paper produced from pretreated wood chips were superior to those from the untreated chips: the basis weight was lower (85.67 and 82.48 g·cm(-2) for paper from untreated KP and HBSP; 79.94 and 80.25 g·cm(-2) for paper from UAE-pretreated KP and HBSP), and the tensile strengths, tearing strengths, bursting strengths, and folding strengths were higher than these of paper after UAE-pretreated, respectively.

How soil fungi function in soil carbon and nutrient cycling is not well understood by using fungal enzymatic differences and their interactions with soil colloids. Eight extracellular enzymes, EEAs (chitinase, carboxymethyl cellulase, β-glucosidase, protease, acid phosphatase, polyphenol oxidase, laccase, and guaiacol oxidase) secreted by ten fungi were compared, and then the fungi that showed low and high enzymatic activity were co-cultured with soil colloids for the purpose of finding fungi-soil interactions. Some fungi (Gomphidius rutilus, Russula integra, Pholiota adiposa, and Geastrum mammosum) secreted 3-4 enzymes with weak activities, while others (Cyathus striatus, Suillus granulate, Phallus impudicus, Collybia dryophila, Agaricus sylvicola, and Lactarius deliciosus) could secret over 5 enzymes with high activities. The differences in these fungi contributed to the alterations of functional groups (stretching bands of O-H, N-H, C-H, C = O, COO- decreased by 11-60%, while P = O, C-O stretching, O-H bending and Si-O-Si stretching increased 9-22%), surface appearance (disappearance of adhesive organic materials), and elemental compositions (11-49% decreases in C1s) in soil colloids. Moreover, more evident changes were generally in high enzymatic fungi (C. striatus) compared with low enzymatic fungi (G. rutilus). Our findings indicate that inter-fungi differences in EEA types and activities might be responsible for physical and chemical changes in soil colloids (the most active component of soil matrix), highlighting the important roles of soil fungi in soil nutrient cycling and functional maintenance.

Concentration of Glomalin Related Soil Protein is reportedly close related to soil functions, but few data is available for GRSP compositional variations and function related to soil properties. In this paper, soils from 0–20 cm, 20–40 cm, 40–60 cm, 60–80 cm, and 80–100 cm layers were collected in 72 poplar shelterbelts in Songnen Plain (6 regions) for implementing this data shortage. GRSP mainly consists of stretching of O–H, N–H, C–H, C=O, COO–, C–O, and Si–O–Si and bending of C–H and O–H. It has seven fluorescent substances of tyrosine-like protein, tryptophan-like protein, fulvic acid-like, humic acid-like, soluble microbial byproduct-like, nitrobenzoxadiazole-like, and calcofluor white-like, with characteristic X-ray diffraction peak at 2θ = 19.8° and 129.3 nm grain size as well as 1.08% low crystallinity. Large spatial variations (intersite and intrasite down profile) were found in either GRSP concentration or these compositional traits. Regression analysis clearly manifested that soil pH should be responsible for these variations. However, negative relations between soil bulk density and GRSP quantity were observed, but not its compositional traits. These basic data in poplar shelterbelt forests are good for understanding the underlying mechanism of GRSP in soil functional maintenance.

An ionic liquids-based ultrasound-assisted extraction (ILUAE) method was successfully developed for extracting shikimic acid from conifer needles. Eleven 1-alkyl-3-methylimidazolium ionic liquids with different cations and anions were investigated and 1-benzyl-3-methylimidazolium bromide solution was selected as the solvent. The conditions for ILUAE, including the ionic liquid concentration, ultrasound power, ultrasound time, and liquid-solid ratio, were optimized. The proposed method had good recovery (99.37%–100.11%) and reproducibility (RSD, n = 6; 3.6%). ILUAE was an efficient, rapid, and simple sample preparation technique that showed high reproducibility. Based on the results, a number of plant species, namely, Picea koraiensis, Picea meyeri, Pinus elliottii, and Pinus banksiana, were identified as among the best resources of shikimic acid.

An efficient microwave-assisted extraction (MAE) technique was employed in simultaneous extraction of luteolin and apigenin from tree peony pod. The MAE procedure was optimized using response surface methodology (RSM) and compared with other conventional extraction techniques of macerate extraction (ME) and heat reflux extraction (HRE). The optimal conditions of MAE were as follows: employing 70% ethanol volume fraction as solvent, soaking time of 4 h, liquid-solid ratio of 10 (mL/g), microwave irradiation power of 265 W, microwave irradiation time of 9.6 min, and 3 extraction cycles. Under the optimal conditions, 151 μg/g luteolin and 104 μg/g apigenin were extracted from the tree peony pod. Compared with ME and HRE, MAE gave the highest extraction efficiency. The antioxidant activities of the extracts obtained by MAE, ME, and HRE were evaluated using a 2,2-di(4-tert-octylphenyl)-1-picrylhydrazyl (DPPH) free radical-scavenging assay, a ferric reducing antioxidant power assay (FRAP), and a reducing power assay. Meanwhile, the structural changes of the unprocessed and processed tree peony pod samples were analyzed by scanning electron microscopy.

An ionic liquids based simultaneous ultrasonic and microwave assisted extraction (ILs-UMAE) method has been proposed for the extraction of rutin (RU), quercetin (QU), from velvetleaf leaves. The influential parameters of the ILs-UMAE were optimized by the single factor and the central composite design (CCD) experiments. A 2.00 M 1-butyl-3-methylimidazolium bromide ([C4mim]Br) was used as the experimental ionic liquid, extraction temperature 60°C, extraction time 12 min, liquid-solid ratio 32 mL/g, microwave power of 534 W, and a fixed ultrasonic power of 50 W. Compared to conventional heating reflux extraction (HRE), the RU and QU extraction yields obtained by ILs-UMAE were, respectively, 5.49 mg/g and 0.27 mg/g, which increased, respectively, 2.01-fold and 2.34-fold with the recoveries that were in the range of 97.62–102.36% for RU and 97.33–102.21% for QU with RSDs lower than 3.2% under the optimized UMAE conditions. In addition, the shorter extraction time was used in ILs-UMAE, compared with HRE. Therefore, ILs-UMAE was a rapid and an efficient method for the extraction of RU and QU from the leaves of velvetleaf.

In Catharanthus roseus, the monoterpene moiety exerts a strong flux control for monoterpene indole alkaloid (MIA) formation. Monoterpene synthesis depends on the methyl-D-erythritol 4-phosphate (MEP) pathway. Here, we have explored the regulation of this pathway in response to developmental and environmental cues and in response to specific enzyme inhibitors. For the MEP pathway entry enzyme 1-deoxy-D-xylulose 5-phosphate synthase (DXS), a new (type I) DXS isoform, CrDXS1, has been cloned, which, in contrast to previous reports on type II CrDXS, was not transcriptionally activated by the transcription factor ORCA3. Regulation of the MEP pathway in response to metabolic perturbations has been explored using the enzyme inhibitors clomazone (precursor of 5-ketochlomazone, inhibitor of DXS) and fosmidomycin (inhibitor of deoxyxylulose 5-phosphate reductoisomerase (DXR)), respectively. Young leaves of non-flowering plants were exposed to both inhibitors, adopting a non-invasive in vivo technique. Transcripts and proteins of DXS (3 isoforms), DXR, and hydroxymethylbutenyl diphosphate synthase (HDS) were monitored, and protein stability was followed in isolated chloroplasts. Transcripts for DXS1 were repressed by both inhibitors, whereas transcripts for DXS2A&B, DXR and HDS increased after clomazone treatment but were barely affected by fosmidomycin treatment. DXS protein accumulated in response to both inhibitors, whereas DXR and HDS proteins were less affected. Fosmidomycin-induced accumulation of DXS protein indicated substantial posttranscriptional regulation. Furthermore, fosmidomycin effectively protected DXR against degradation in planta and in isolated chloroplasts. Thus our results suggest that DXR protein stability may be affected by substrate binding. In summary, the present results provide novel insight into the regulation of DXS expression in C. roseus in response to MEP-pathway perturbation.

Limited data are available on the ectomycorrhizae-induced changes in surface structure and composition of soil colloids, the most active portion in soil matrix, although such data may benefit the understanding of mycorrhizal-aided soil improvements. By using ectomycorrhizae (Gomphidius viscidus) and soil colloids from dark brown forest soil (a good loam) and saline-alkali soil (heavily degraded soil), we tried to approach the changes here. For the good loam either from the surface or deep soils, the fungus treatment induced physical absorption of covering materials on colloid surface with nonsignificant increases in soil particle size (P>0.05). These increased the amount of variable functional groups (O–H stretching and bending, C–H stretching, C=O stretching, etc.) by 3–26% and the crystallinity of variable soil minerals (kaolinite, hydromica, and quartz) by 40–300%. However, the fungus treatment of saline-alkali soil obviously differed from the dark brown forest soil. There were 12–35% decreases in most functional groups, 15–55% decreases in crystallinity of most soil minerals but general increases in their grain size, and significant increases in soil particle size (P

Ultrasound was used for the extraction of larch arabinogalactan from Larix gmelinii. The optimal conditions for ultrasound extraction were determined by response surface methodology. Specifically, the Box-Behnken design was employed to evaluate the effects of three independent variables: ultrasound time, temperature, and liquid-solid ratio. The highest arabinogalactan yield (11.18%) was obtained under the optimal extraction condition (extraction temperature 41.5°C, extraction time 24.3 min, and liquid-solid ratio 40 mL/g). In addition, the antioxidant activity of arabinogalactan that was extracted from dihydroquercetin extraction residues exhibited a moderate and concentration-dependent hydroxyl radical-scavenging capacity, ferric-reducing power, and reducing power. The wood material was characterized before and after processing by scanning electron microscopy and Fourier-transform infrared spectroscopy.

An ionic liquid-based ultrasonic-assisted extraction method has been developed for the effective extraction of procyanidins from Larix gmelinii bark. So as to evaluate the performance of ionic liquids in ultrasonic-assisted extraction process, the effects caused by changes in the anion and the alkyl chain length of the cation on the extraction efficiency were investigated in this paper. The results indicated that the characteristics of anions had remarkable effects on the extraction efficiency of procyanidins, and 1-butyl-3-methylimidazolium bromide ([Bmim]Br) aqueous solution was the best among the investigated ionic liquids. The optimum conditions for the extraction were as follows: [Bmim]Br concentration 1.25 M, soak time 3 h, solid-liquid ratio 1 : 10, ultrasonic power 150 W, and ultrasonic time 30 min. This work not only introduces a simple, green, and highly efficient sample preparation method for extraction of procyanidins from L. gmelinii bark, but also reveals the tremendous application potential of ionic liquids.

Thinning management is used to improve timber production, but only a few data are available on how it influences ecosystem C sink capacity. This study aims to clarify the effects of thinning on C sinks of larch plantations, the most widespread forests in Northeastern China. Both C influx from biomass production and C efflux from each soil respiration component and its temperature sensitivity were determined for scaling-up ecosystem C sink estimation: microbial composition is measured for clarifying mechanism for respiratory changes from thinning treatment. Thinning management induced 6.23 mol C m−2 yr−1 increase in biomass C, while the decrease in heterotrophic respiration (Rh) at the thinned sites (0.9 mol C m−2 yr−1) has enhanced 14% of this biomass C increase. This decrease in Rh was a sum of the 42% decrease (4.1 mol C m−2 yr−1) in litter respiration and 3.2 mol C m−2 yr−1 more CO2 efflux from mineral soil in thinned sites compared with unthinned control. Increases in temperature, temperature sensitivity, alteration of litters, and microbial composition may be responsible for the contrary changes in Rh from mineral soil and litter respiration, respectively. These findings manifested that thinning management of larch plantations could enhance biomass accumulation and decrease respiratory efflux from soil, which resulted in the effectiveness improvement in sequestrating C in forest ecosystems.

An ionic liquid-based ultrasonic/microwave-assisted simultaneous distillation and extraction (IL-UMASDE) method for isolating camptothecin (CPT), 10-hydroxycamptothecin (HCPT), vincoside-lactam (VCS-LT), and essential oils (EOs) from Camptotheca acuminata Decne fruits was developed. The important parameters were optimized using single-factor and central composite design experiments. The optimum conditions were 0.75 M 1-octyl-3-methylimidazolium ([C8mim]Br) as the extraction solvent, a liquid–solid ratio of 13.7 mL/g, an extraction time of 33.2 min, a microwave power of 582 W, and a fixed ultrasonic power of 50 W. The yields of CPT, HCPT, and VCS-LT obtained under the optimum conditions were 2.463, 0.164, and 0.297 mg/g, respectively; these are 1.08-, 1.12-, and 1.04-fold higher, respectively, than those obtained by conventional 55% ethanol heat reflux extraction (HRE). The extraction time for the equilibrium yields of CPT, HCPT and VCS-LT using IL-UMASDE was 33.6%, 58.5%, 63.1%, and 66.8%, respectively, less than the corresponding times using IL-MASDE, IL-ultrasonic-assisted extraction (IL-UAE), 55% ethanol UAE and 55% ethanol HRE. The yield of EOs obtained using IL-UMASDE was 0.793 mg/g, i.e., 1.31-fold higher than that obtained by conventional hydrodistillation extraction (HDE). The components of the EOs obtained using IL-UMASDE and HDE were similar. The extraction time for the equilibrium yields of EOs using IL-UMASDE is 33.6%, 58.5%, 52.6%, and 72.3% lower than those for IL-MASDE, water-UMASDE, water-MASDE, and HDE, respectively. Compared with other extraction methods, IL-UMASDE gave the highest yields of CPT, HCPT, VCS-LT, and EOs and also had the shortest extraction time. IL-UMASDE is a potential green and highly efficient technique for the extraction of CPT, HCPT, VCS-LT, and EOs from Camptotheca acuminata Decne fruits.

In the present study, a method for the separation by macroporous resins of 10-hydroxycamptothecin (HCPT) from the extracts remaining from the production of camptothecin (CPT) has been developed. The adsorption/desorption characteristics of HCPT on eight macroporous resins were compared. Of these, the X-5 resin offered the best adsorption/desorption densities for HCPT, with its adsorption data being best fitted by the Langmuir and Freundlich isotherms. Dynamic adsorption/desorption experiments were carried out in a column packed with X-5 resin in order to optimize the separation process. The optimum parameters for adsorption were as follows: sample solution concentration = 0.3156 mg/mℓ, processing volume = 5 BV (bed volume), flow rate = 4 BV/h, temperature = 313 K, pH = 5; for desorption, the corresponding parameters were: 10–60% ethanol as eluent, processing volume = 2 BV, flow rate = 4 BV/h. After being treated with X-5 resin, the content of HCPT in the dry product increased from 0.30% to 7.19%, i.e. a 23.9-fold increase when compared to the untreated extract, with the recovery yield of HCPT being 82.1%.