The methanolic (MeOH) extract of A. fruticosa bark, which showed immune-regulatory activities, was separated to purify an active compared by means of a multi-stage column chromatography. This resulted in the isolation and characterization of an isoflavone glycoside named 4', 6-Dimethoxyisoflavone-7-O-β-D-glucopyranoside. Immuno-regulatory activities of the crude extract of Amorpha fruticosa LINNE bark were compared with that of an isoflavone glycoside (4', 6-dimethoxyisoflavone-7-O-β-D-glucopyranoside). The crude methanolic extract of A. fruticosa and purified single compound showed 16% of relatively low cytotoxicity at a maximum concentration of 1.0 g/L in cultivated normal human lung cell line (HEL299). Cell growth of human T cells was increased up to 15%, 0.5 g/L of the crude extract added group. This was higher than a single compound added one. On the other hand, specific production rates of IL-6 and TNF-α from T cell were higher in the purified compound treat group (0.82×10-4 pg/cell and 1.08×10-4 pg/cell, respectively), compared to 0.5 g/L of the crude extract added group (0.65×10-4 pg/cell and 0.84×10-4 pg/cell, respectively). In addition, the growth of NK-92MI cells incubated with the crude extract was higher up to 56% over the cells grown with a single compound (0.5 g/L). In overall, the crude extract showed relatively higher immuno-regulatory activities compared with a single compound, probably due to the synergic effect given by other substances existed in the crude extract. Even though the siolated compound stimulated higher secretion of cytokines from human T cells.
Paeoniae radix has been widely used for its anti-allergic, anti-inflammatory and analgesic effects, and demonstrated to have anticonvulsant, memory enhancing and anxiolytic activities. The present study was performed to examine the protective effect of methanol extract of Paeoniae radix (PR) from Paeoniae Japonica Miyabe et Takeda (Paeoniaceae) on hydrogen peroxide (H2O2)-induced neurotoxicity using cultured rat cerebral cortical neuron. H2O2 produced a concentration-dependent reduction of neuronal viability, PR, over a concentration range of 10 to 100 μg/ml showed concentration-dependent decrease of the H2O2(100 μM)-induced neuronal cell death, as assessed by a 3-[4,5-dimethylthiazol-2-yl]-2,5-di-phenyl-tetrazolium bromide assay and the number of apoptotic nuclei, evidenced by Hoechst 33342 staining. PR (100 μg/ml inhibited 100 μM H2O2-induced elevation of the cytosolic Ca2+ concentration ([Ca2+]c), which was measured by a fluorescent dye, flue-4 AM. PR (50 μg/ml inhibited glutamate release into medium induced by 100 μM H2O2, which was measured by HPLC, and generation of reactive oxygen species (ROS). These results suggest that PR may mitigate the H2O2-induced neurotoxiciy by interfering with the increase of [Ca2+]c, and then inhibiting glutamate release and generation of ROS in cultured neurons.
As a result of cytotoxic compounds against cancer cell lines from natural sources, senven compounds were isolated from the leaf and twig of Acer okamotoanum Nakai. The compounds (1-7) were identified as ethyl gallate (1), methyl gallate (2), gallic acid (3), trans resveratrol-3-O-β-D-glucopyranoside (4), acertannin (5), nikoenoside (6), and fraxin (7) by physicochemical and spectroscopic data (including mp, UV, IR, MS, 1H-NMR, 13C-NMR, DEPT, and HMBC) in comparison with those of published papers. All the compounds were tested for their cytotoxic activity against L1210, HL-60, K562, and B16F10 cancer cell lines in vitro by MTT assay method. Compounds 1-3 and 5 showed cytotoxic activity against all tested cell lines with IC50 values ranged from 12.5 to 72.2 μM. Of the compounds, methyl gallate (2) exhibited the most potent cytotoxic activity against L1210, HL-60, K562, and B16F10 tumor cells with IC50 values of 12.5, 48.3, 22.8, and 22.8 μM, respectively. Other compounds did not show any cytotoxic activity against four cancer cell lines.
Antioxidant activity of the extract fractions from leaves, stems, roots and flowers of Cirsium pendulum Fisch. was investigated. The results showed the greatest antioxidant activities in leaves by Rancimat, TBA and DPPH methods. Extracts of common thistle plants dose-dependently increased DPPH free radical scavenging activity, The extract from flowers and its hexane fraction showed the strongest antioxidant activity. HPLC analysis showed that BuOH fraction of the leaves had the highest amount of antioxidant chlorogenic and p-coumaric acids at 5.38 and 9.71 mg 100 g-1, respectively. It implies that common thistle plants had potent antioxidant activity, and their activities were differently exhibited depending on plant part and solvent fraction.
An efficient plant regeneration of C. asiatica was achieved from organogenesis using petiole explants of in vitro plantlet on MS basal medium controled with different plant growth regulators (NAA,2,4-D, IAA kinetin, and BA). Best results that 50%, efficiency of regeneration per explant for regeneration were obtained with IAA 17.13 μM and BA 8.9 μM. Formation of adventitious shoots via organogenesis from the petiole explant was verified by histological sectioning of plantlets. Regenerated plants were transplanted into soil.
A method for plant regeneration via organogenesis from Pelagonium inquinans leaf disc has been developed. Mature leaf explants were collected from field-grown plants and used for the induction of adventitious shoot regeneration on Murashige and Skoog (MS) medium supplemented with 3% (w/v) sucrose plus plant growth regulators. Maximum shoot organogenesis, with 11.8±1.5 shoots (98.6%) per leaf disc, was obtained with 2 mg/l N6-benzyladenine (BA) and 0.5 mg/l α-naphthyleneacetic acid (NAA) in 30 days. For rooting, the in vitro proliferated and elongated shoots were excised into 1.5-2 cm in length microcutting, which were plated individually on an half-strength MS (1/2MS) medium supplemented with 2% (w/v) sucrose plus various concentrations of indole-3-butyric acid (IBA). Shoots rooted with a frequency of 100% following culture on 1/2MS medium containing 0.5 mg/l IBA.
Regions of allelic loss on chromosomes in many tumors of human and some experimental animals are generally considered to harbor tumor-suppressor genes involved in tumorigenesis. Allelotype analyses have greatly improved our under-standing of the molecular mechanism of radiation lymphomagenesis. Previously, we and others found frequent loss of heterozygosity (LOH) on chromosomes 4, 11, 12, 16 and 19 in radiation-induced lymphomas from several F1, hybrid mice. To examine possible contributions of individual tumor-suppressor genes to tumorigenesis in p53 heterozygous deficiency, we investigated the genome-wide distribution and status of LOH in radiation-induced lymphomas from F1 mice with different p53 status. In this study, we found frequent LOH (more than 20%) on chromosomes 4 and 12 and on chromosomes 11, 12, 16 and 19 in radiation-induced lymphomas from (STS/A×MSM/Ms)F1 mice and (STS/A×MSM/Ms)F1-p53KO/+ mice, respectively. Low incidences of LOH (10-20%) were also observed on chromosomes 11 in mice with wild-type p53, and chromosomes 1, 2, 9, 17 and X in p53 heterozygous-deficient mice. The frequency of LOH on chromosomes 9 and 11 increased in the (STS/A×MSM/Ms)F1-p53KO/+ mice. Preferential losses of the STS-derived allele on chromosome 9 and wild-type p53 allele on chromosome 11 were also found in the p53 heterozygous-deficient mice. Thus, the putative tumor-suppressor gene regions responsible for lymphomagenesis might considerably differ due to the p53 status.
We established a practical method for rapid and large-scale production of Valeriana fauriei var. dasycarpa Hara roots by bioreactor culture and confirmed valerenic acids and valepotriates production. We also compared valerenic acids and valepotriates production patterns according to various media conditions. Among the media tested, B5 medium gave the maximum biomass production of 101 g fresh weight, which was a 5.03-fold multiplication rate obtained 4 weeks after inoculation of 20 g of fresh weight. The best production of total valerenic acids (7.86 mg/l) and valepotriates (8.96 mg/l) was B5 medium.
Two Atractylodes species, A. japonica Koidz. ex Kitam. (AJ) and A. macrocephala Koidz (AM) were used in this study. AJ population had higher amounts of Sesquiterpenoids and stronger tolerance to root rot but less vigor of root growth than AM population. Two populations (AJ and AM) were crossed to make interspecific hybrid population. A total of 98 lines propagated clonally were selected from a cross of AJ and AM, and evaluated for contents of sesquiterpenoids, atractylon (ATLN) and atractylenolide III (AT3) using high performance liquid chromatography (HPLC), and growth characters such as plant height, stem number and root weight. HPLC profiles of the hybrids were compared with those of parent plants, and it demonstrated the production of introgression hybrid by crossing between AJ and AM. Of 98 clonal lines,10 lines were selected by 10% level based on the growth vigor and tolerance to root rot, and AJM2102-51 line showed the heaviest root weight (117.1 g/plant) among them. A total of 98 hybrid lines contained on average 0.16 ± 0.10 mg/g of AT3, 2.00 ± 1.37 mg/g of ATLN, and 2.16 ± 1.40 mg/g of total sesquiterpenoids, showing high coefficients of variation (above 65%). Ten lines having high contents of sesquiterpenoids were selected, and AJM2101-15 had the highest amount (9.83 mg/g) of ATLN, and showed 40.8 g/plant of root weight similar to mean value (39.9 g/plant) of hybrid lines. The result showed that the introgression of both characters of vigorous growth from AM and high sesquiterpenoids content from AJ could be possible to make new hybrid lines by crossing between AJ and AM.
This study was carried out to investigate spectral irradiance characteristics of blue, yellow, and blue-black colored polyethylene (PE) shading net and the effect on growth characteristics and yield in ginseng seedling. The spectral irradiance (μmol/m2/s/nm) showed the peak at 498 nm in both of blue and blue-black PE shading net, and 606 nm under yellow PE one. The intensity of blue light in blue shading was more strong than that of blue-black shading, control. Blue shading was increased by 17% and 23% in accumulated quantum for daytime, 0.5℃ and 0.2℃ in maximum temperature on June 2 than that of yellow and blue-black shading, respectively, but heat injury ratio of the former was lower than that of the latter. Chlorophyll content and stem length in blue shading were decreased more significantly than those of yellow and blue-black shading. The specific leaf weight was higher under blue and yellow shading than that of blue-black shading. Ginseng seedling harvested in blue shading was increased by 13~17% in the number of root, and 17~20% in root weight per m 2 compared to yellow and blue-black shading owing to the increase of survived plant, and the decrease of specific leaf weight, heat injury ratio, and stem length.
The purpose is to study the identification method of Huangjinju powder for injection and the medicinal materials by the fingerprint off-ray Diffraction Fourier (XRDF). We used the same method on both the studying of Huangjinju and the medicinal materials. Then we selected a few components alignment to compare. We analyzed the data by setting up the deviation d(a) as ±0.05 to calculate the rate of special mark on the sample (Px) and on the patent (P). The special XRDF of Huangjinju[d(a)/(I/I0)] have 5 peaks that have not expressed in medicinal materials. Therefore Px is 22.73%. Flos Trollii Chinensis has 3 special marks and Px is 17.65%. Flos Chrysanthemi Indici has 1 special mark and Px is 3.57%. Its coincided interplanar spacing with the patent is 2.907a. Flos Lonicerae Japonicae has 6 special marks and Px is 23.08%. Its special mark in the patent are 4.95/14 and 4.50/15, respectively. The P is 9.09%. Its coincided interplanar spacing with the patent is 2.910a and 3.05a, respectively. The number of special XRDF mark peaks of baicalin is 9 and Px is 18.37%. Its coincided interplanar spacing with the patent is 2.910a. It has visible mark and specificity adopting XRDF fingerprint to identify Huangjinju and medicinal materials. Establishing the quality standard is a synthetic index that depends both on special marks in the medicinal materials of the patent and on the coincidence peak data.