Background : Licorice has been used as a source of medicine and a food material in East-Asia. Recently, demand for licorice increased in market due to a growing interest in health. Thus we conducted breeding research to solve the problems associated with domestically cultivated licorice such as low productivity and low glycyrrhizin content. Methods and Results : We crossed European licorice (G. glabra L.; female parent) and Chinese licorice (G. uralensis Fisch; male parent) in the greenhouse in May 2007. In September 2007, crossed and germinated seeds were retrieved and sown in the greenhouse. In June 2008, stolons were separated from the F1 licorice seedlings and cultivated, resulting in 32 clonal lines of interspecific hybrids. Among them we selected good lines and then conducted the replicated yield trials (RYT) in 2012-2013 and local adaptability test (LAT) in 2014-2015. The results, GLYES9 showed that was elect of stem, oblong of leaf shape, red-brown of root color. Glycyrrhizin conten of GLYES9 (3.0%) was higher than G. uralensis (1.9%) at four regions from 2014 to 2015. GLYES9 was less than 10% in the desease of brown spot (G. uralensis was more than 30%). The root yield of GLYES9 was 4.31 ton per hectare, which was increased 193% compared with a check variety of G. uralensis. Therfore, we named GLYES9 as new cultivar ‘Dagam’. Conclusion : Depending on the above results, we have developed a new licorice cultivar ‘Dagam’ by the medicinal crop breeding team of National Institute of Horticulture and Herbal Science, RDA, in 2015. It showed brown spot disease resistant, high-glycyrrhizn content and high-yielding than colleted Glycyrrhiza spp.

Background: This study was performed to evaluate the protective effect of Saururus chinensis ethanol extract (SCE) against styrene toxicity in mouse spermatocyte cells [GC-2spd (ts) cell line]. Methods and Results: Cytotoxicity in mouse spermatocyte cells was measured using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Generation of reactive oxygen species (ROS) was determined using 2’,7’-dichlorodihydrofluorescein diacetate (DCF-DA) assay. Semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) and western blotting were performed to quantify the mRNA and protein expression levels, resepectiviely, of stress or apoptosis-related genes including p21, p53, heat shock protein 70 (Hsp70), heat shock protein 90 (Hsp90), Bax, Bcl-2, and caspase-3. The results of the MTT assay showed that 50 ㎍/㎖ SCE did not affect cell viability. ROS generation in mouse spermatocyte cells increased by treatment with 100 μM styrene, and decreased by co-treatment with SCE. SCE repressed the mRNA expression of stress-related genes, which increased by styrene treatment. In addition, SCE inhibited the apoptosis of mouse spermatocyte cells by ameliorating mRNA and protein levels of apoptotic genes that were altered by styrene treatment. Conclusions: These results suggest that SCE may alleviate styrene toxicity in mouse spermatocyte cells by reducing ROS stress and regulating genes related to styrene toxicity.

Background: The study was conducted to elucidate the extraction conditions under which white ginseng has cognition-improving efficacy.Methods and Results: Extracts from white ginseng under different solvent and temperature conditions were analyzed for ginsenoside content and inhibitory effect on N-methyl-D-aspartate (NMDA) receptor and acetylcholinesterase. The total ginsenoside contents and amounts of ginsenoside Rb1 plus ginsenoside Rg1 from the 1st extracts (prepared with EtOH/H2O as solvent) were higher than those from the 2nd extracts (extracted with H2O after the 1st EtOH/H2O extraction). The contents in the 1st and 2nd extracts produced at 80°C were also higher than those obtained at 50°C. Samples from the 1st extraction at 80°C indicated higher inhibitory activities on NMDA receptors-whose excessive activation is thought to mediate the calcium-dependent neurotoxicity associated with several neurodegenerative diseases-than those from the 2nd extraction. Among the samples prepared at varying temperatures, the extract prepared at 50°C showed the highest suppression activity on NMDA receptors. Note, however, that the extracts from the 2nd extraction at 50°C inhibited acetylcholinesterase-whose inhibition could be a therapeutic strategy for neurodegenerative diseases with cognitive deficits and memory malfunction-more effectively than those from the 1st extraction.Conclusions: To enhance the cognition-improving activity of white ginseng extract, it is suggested that the extracts be utilized after being combined the 1st extracts (made with EtOH/H2O solvent) and the 2nd extracts (prepared with H2O) at low temperature.

Background : Acanthopanax sessiliflorus (Rupr. et Maxim) Seem, belonging to the Araliaceae family, is widely distributed in Korea, China, and Japan. The plants belonging to Acanthopanax species are traditionally used in Korea as anti-rheumatoid arthritis, anti-inflammatory and anti-diabetic drugs and are recognized to have ginseng-like activities. A simple and sensitive high-performance liquid chromatographic (HPLC) method was developed and validated for independent analysis of major compounds and chlorogenic acid in A. sessiliflorus fruits. Chlorogenic acid was reported that prevent cancer and cardiovascular disease in vivo. Also, it has antioxidant effect in vitro test. In the previous experiment, chlorogenic acid were found in A. sessiliflorus fruits. This study was performed to identification of the major compounds and investigate the method validation for the determination of chlorogenic acid in A. sessiliflorus fruits. Methods and Results : Three major compounds were recorded on a Varian Unity Inova AS-400 FT-NMR spectrometer and analyzed by the new HPLC analysis method. HPLC analysis was carried out using an Waters e2695 and PDA detector. The new analyasis method was validated by the measurement of intra-day, inter-day precision, accuracy, limit of detection (LOD, S/N=3), and limit of quantification (LOQ, S/N=10) of chlorogenic acid. The results showed that the correlation coefficient (R2) for the calibration curves of chlorogenic acid was 0.997 in terms of linearity. The limit of detection (LOD) and limit of quantification (LOQ) were 0.565 ㎍/ml and 2.88 ㎍/ml, respectively. There was no interfering peak observed each other and HPLC system was suitable for analysis showing goodness of peak and high precision. Conclusion : This method is suitable to detect and quantify major compounds in A. sessiliflorus fruits. Furthermore, the result will be applied to establish chlorogenic acid as an standard compound for A. sessiliflorus fruits.

Background : Cynanchum wilfordii and Cynanchum auriculatum belong to the Asclepiadaceae family and appear morphologically similar. In order to discriminate them, it is needed to find the presence of sap and the leaf shapes: C. auriculatum has a blade ovate leaf comparing to C. wilfordii. However, in the herbal medicine market, they have been handled as cut and dried roots. Due to their similar morphology, it is limited to distinguish the roots of C. wilfordii and C. auriculatum. Recently in Korea, it has been a critical issue to misuse these two roots in the herbal market and food industry. Thus, it is required to establish a robust tool for the discrimination and quality control of them. Methods and Results : To separation and characterization of flavor compounds, C. wilfordii and C. auriculatum samples were analyzed by head space solid phase micro extraction (HSS) fiber coupled with gas chromatography-mass spectrometry using Rtx-5MS (30 m x 0.25 mm x 0.25 μm) column. As a result, We have identified compounds of a few hundred in aliphatic aldehydes and aliphatic alcohols, alkenes and acids, aromatic compounds, aromatic compounds containing nitrogen & sulfur, etc,. In particular, The aliphatic and aromatic compounds had been clearly separated on the second dimensional direction by using two-dimensional GC. Conclusion : The volatile flavor compounds of C. wilfordii and C. auriculatum could easily analyzed without pre-treatment with improved resolution and sensitivity using HSS-GCxGC-TOFMS. We have identified compounds of a few hundred in C. wilfordii and C. auriculatum sample. And It was more accurately qualitative confirmed with separation of GCxGC and TSD. We have confirmed the PCA and PLS-DA Plot that was classified depending on C. wilfordii and C. auriculatum through multivariate statistical analysis of the identified flavor compounds.

Background : Allergy is a common disease caused by type I hypersensitivity reaction of the immune system. Unfortunately, there is no proper treatment for allergy. Therefore, the discovery of therapeutic drugs for allergy is essential. In this study, the crude extracts of 56 plant parts were screened for anti-allergy effects in RBL-2H3 cells. Methods and Results : IgE-sensitized RBL-2H3 cells were individually treated with 56 extracts of medicinal herbs at the final concentration of 20 ㎍/㎖ and stimulated with the antigen DNP-BSA. β-Hexosaminidase release, interleukin-4 (IL-4) secretion, and cell viability in the sample treated cells were measured. Among the tested samples, extracts from the root of Polygonatum stenophyllum Maxim., aerial part of Acer triflorum Kom., and leaf of Pyrus pyrifolia var. culta (Makino) Nakai showed inhibitory effects on β-hexosaminidase release. The aerial part of Peucedanum japonicum Thunberg and seed of Panax ginseng C.A.Mey. showed suppressive activities on IL-4 secretion. All of the extracts were not cytotoxic at the tested concentration. Conclusion : From the result, six extracts including Polygonatum stenophyllum Maxim. (root) and Acer triflorum Kom. (aerial part) inhibited both β-hexosaminidase release and IL-4 secretion in IgE-sensitized RBL-2H3 cells. The use of these extracts for developing anti-allergy materials is suggested.

Background : Five medicinal herbs have been selected from the preliminary screening for in vitro anti-allergy activity (in RBL-2H3 cells). The present study is conducted to investigate the inhibitory effects of the medicinal herbs on allergic inflammation in other kind of cells. Methods and Results : Cells treated with five extracts prepared from Betula costata Trautv. (aerial part), Camellia sinensis L. (aerial part), Polygonatum stenophyllum Maxim. (root), Pyrus pyrifolia var. culta (Makino) Nakai (leaf), and Rehmannia glutinosa (Gaertn.) Libosch. ex Steud. (root) were measured for mRNA levels of TNF-α on HaCaT keratinocytes stimulated by TNF-α /INF-γ and for mRNA levels of IL-2 in Jurkat T cells mediated by PMA/A23187. Pre-treatment with the five extracts reduced the mRNA levels of TNF-α in HaCaT cells and mRNA levels of IL-2 in Jurkat T cells. In particular, the leaf extract of Pyrus pyrifolia var. culta (Makino) Nakai significantly and dose-dependently decreased the mRNA levels of TNF-α and IL-2. To determine the toxicity of the extracts from the selected medicinal herbs to HaCaT cells and Jurkat T cells, the viabilities of the cells treated with several concentrations of the five extracts were measured by MTT assay. Extracts of Polygonatum stenophyllum Maxim. (root), Pyrus pyrifolia var. culta (Makino) Nakai (leaf), and Rehmannia glutinosa (Gaertn.) Libosch. ex Steud. (root) (up to 250 ㎍/㎖) did not show cytotoxic effects on HaCaT cells and Jurkat T cells. On the other hand, 250 ㎍/㎖ of extracts of Betula costata Trautv. (aerial part) and Camellia sinensis L. (aerial part) reduced cell viability in both cells. Conclusion : These results demonstrate that the leaf extract of Pyrus pyrifolia var. culta (Makino) Nakai has an anti-inflammatory effect by inhibiting pro-inflammatory cytokine expression. Therefore, the leaf of Pyrus pyrifolia var. culta (Makino) Nakai can be a useful resource for the development of anti-allergy/anti-inflammatory materials.

Background : Ginseng, one of most famous traditional oriental medicines, has been known for a number of pharmacological properties including anti-tumor, anti-diabetic, anti-fatigue, anti-stress, anti-oxidative, and anti-aging effects. 20(S)-Protopanaxadiol (PPD), a intestinal metabolite of ginsenosides, is one of the active ingredients in ginseng. In this study, we investigated the synergistic anticancer effect of 20(S)-PPD and temozolomide (TMZ) and the mechanism of 20(S)-PPD on glioblastoma cells. Methods and Results : We examined cell viability and the morphological changes of C6 cells after treatment of 20(S)-PPD and TMZ. 20(S)-PPD showed a potent antiproliferative activity against C6 cells by triggering apoptosis. 20(S)-PPD-induced apoptosis was characterized by a dose-dependent mitochondrial damage. 20(S)-PPD and TMZ had a synergistic effect in increasing mitochondrial damage via caspase 3 activation. Conclusion : These results revealed an unexpected mechanism of 20(S)-PPD and TMZ, triggering a mitochondrial-mediated apoptosis in C6 cells. Our findings encourage further studies of 20(S)-PPD as a promising chemopreventive agent against glioblastoma.

Background : Recently, ginseng (Panax ginseng C.A. Meyer.) berry has been used as a health-promoting supplements. Also, Mulberries (Morus alba L.) fruit have been used in traditional herbal medicine to treat and prevent diabetes. In this study, we measured the cytotoxicity after fermentation of the extracts in Panax Ginseng Berry and Mulberry Fruit. Methods and Results : The extracts were prepared by decoction for 3 hours in distilled water (100 g/L). The dried extract was then dissolved in phosphate-buffered saline (PBS) in preparation for use. Cell viability was examined by an MTT assay. RAW 264.7 cells were seeded at 1 × 104/mL densities in 96-well plates. Each grouping had a non-treated group as the control. The extracts were added to each well and incubated for 24 hours at 37°C and 5% CO2. The MTT solutions (5 mg/mL) were added to each well, and the cells were cultured for another 2 hours. The supernatant was then discarded, and 100 μL of dimethyl sulfoxide was added to each well. The optical density was read at 540 nm. Conclusion : Probiotics and prebiotics modulate the composition of human and domestic animal gut microbiota. The beneficial effects may result from suppression of harmful microorganisms or stimulation of organisms which contribute in a positive way to the nutrition and health of human and domestic animal. Recently, fermentation using microorganisms for the production of more effective compounds has been extensively studied. In particular, the novel pharmacological effects of a new compound generated by fermentation have been reported. Some previous studies have demonstrated that Fermented herbal medicine extract showed better bioactivity than normal herbal Plants extract when used at the same concentration.

Background: The public has increasing concerns about herbal crops owing to insufficient information on biological hazards such as foodborne pathogens. Therefore, the objective of this study is the development of a herbal crop quality control system through monitoring with biological hazard analysis. Today, it is estimated that millions of people become ill every year from food contamination. The public demands agricultural products of stable and consistent quality. Governments have the responsibility of establishing the standards, legislation and enforcement programs necessary to control food quality and safety. However, research on the biosafety of herbal crop products is still insufficient. Therefore, the implementation of monitoring systems with high standards is critical for public safety. Methods and Results: In this study, we collected 52 samples of herbal crop products, and conducted both quantitative and qualitative biological hazard analysis. With biological hazard analysis, aerobic bacteria, Staphylococcus aureus, Salmonella spp., Escherichia coli, Coliforms, and Listeria spp. could be detected. Conclusions: Herbal crops were found to be contaminated with aerobic bacteria at 3.69 ± 0.32 log CFU/g. Staphylococcus aureus, Salmonella spp., Escherichia coli, Coliforms, and Listeria spp. were not detected in any of the samples. This research suggests that continuous monitoring of biological hazards is required to improve the quality of herbal crops.

Plant breeding requires the collection of genetically diverse genetic resources. Studies on the characteristics of Platycodon grandiflorum resources have not been carried out so far. The present study was carried out to discriminate P. grandiflorum based on morphological characteristics and genetic diversity using simple sequence repeat (SSR) markers. Methods and Results :We collected 11 P. grandiflorum cultivars: Maries II, Hakone double white, Hakone double blue, Fuji white, Fuji pink, Fuji blue, Astra white, Astra pink, Astra blue, Astra semi-double blue and Jangbaek. Analyses of the morphological characteristics of the collection were conducted for aerial parts (flower, stem and leaf) and underground parts (root). Next, the genetic diversity of all P. grandiflorum resources was analyzed using SSR markers employing the DNA fragment analysis method. We determined that the 11 P. grandiflorum cultivars analyzed could be classified by plant length, leaf number and root characteristic. Based on the genetic diversity analysis, these cultivars were classified into four distinct groups. Conclusions : These findings could be used for further research on cultivar development using molecular breeding techniques and for conservation of the genetic diversity of P. grandiflorum. Moreover, the markers could be used for genetic mapping of the plant and marker-assisted selection for crop breeding.

A new extraction method-heated ultrasonic extraction was qualitatively and quantitatively analyzed for the extraction of major ginsenosides from ginseng extract; this new high-performance liquid chromatography (HPLC) method was compared with the official extraction method of Korean industrial standards and standard for health functional food. Methods and Results : Ginsenoside compounds were analyzed for 35 minutes by the new HPLC analysis method using a Halo® RP-Amide column. The new HPLC analysis method was validated by the measurement of intra-day and inter-day precision, accuracy, limit of detection (LOD), and limit of quantification (LOQ) of each ginsenoside. The correlation coefficients (r2) for the calibration curves of the ginsenoside compounds were over 0.9997 in terms of linearity. The heated ultrasonic extraction method using ultrasonication for 30 minutes at 50℃ yielded higher amount of ginsenosides than the extraction method of the Korean industrial standards owing to the enhancement of extraction efficiency. Conclusions : Compared to the other extraction methods, the heated ultrasonic extraction method yielded a higher amount of ginsenoside Rb1 than Rg1 index compounds for the quality evaluation of ginseng roots.

Background : This study was conducted to determine out the effect of storage temperature on the quality of fresh ginseng (Panax ginseng C. A. Meyer) during distribution. Methods and Results : Fresh ginseng was washed, packed in 30㎛ low density polyethylene (LDPE) film, then stored at 0, −2 and −4℃. After 4 weeks of storage, ginseng was then stored at 5℃, as a simulation of the distribution process. Ginseng stored at −4℃ showed higher respiration rate, ethylene production and electrolyte conductivity during the distribution phase than those stored at 0 and −2℃. Decay and browning rate rapidly increased following 3 weeks of distribution in samples stored −4℃. However ginseng stored −2℃, which is below freezing point, for 4 weeks did not show the physiological change or quality deterioration. Ginsenoside contents decreased during storage for all plant, but did not differ significantly between storage temperatures. Conclusions : Storage at temperatures below −2℃ can negatively affect respiratory characteristics and electrolyte leakage and increase quality deterioration and decay rates during distribution.