Aralia cordata (A. cordata), which belongs to Araliaceae, is a perennial herb widely distributed in East Asia. We evaluated the anti-inflammatory effect of stems (AC-S), roots (AC-R) and leaves (AC-L) extracted with 100% methanol of A. cordata and elucidated the potential signaling pathway in LPS-stimulated RAW264.7 cells. The AC-L showed a strong anti-inflammatory activity through inhibition of NO production. AC-L dose-dependently inhibited NO production by suppressing iNOS, COX-2 and IL-β expression in LPS-stimulated RAW264.7 cells. AC-L inhibited the degradation and phosphorylation of IκB-α, which donated to the inhibition of p65 nuclear accumulation and NF-κB activation. Furthermore, AC-L suppressed the phosphorylation of ERK1/2 and p38. These results suggested that AC-L may utilize anti-inflammatory activity by blocking NF-κB and MAPK signaling pathway and indicated that the AC-L can be used as a natural anti-inflammatory drugs.

본 연구는 완주에서 재배된 세계 주요 11개 적포도 품종으로 제조된 적포도주의 향기 성분을 headspace-solid phase microextraction 분석법으로 확인하였다. 향기성분은 총 75종이 확인되었다. 아로마화합물은 그들의 OAV 값에 의해 5 그룹으로 나뉘었다. 알콜, 알데하이드, 에스테르, C6 화합물이 11개 적포도주의 주요한 향기성분이었다. Isoamy alcohol 알콜과 phenylethyl 알콜은 11개 포도주에서 공통적으로 꽃향기, 달콤한 향을 나타내는데 중요한 물질이었다. Octanoic acid, ethyl ester, hexanoic acid ethylester은 모든 레드와인에서 과실향과 꽃향, 달콤한 향을 내는 중요한 성분이었다. 1-Hexanol은 모든 포도주에서 분석되었으나 풀향을 나타내는 향으로 나타났다. Chanceller, Malbec, marchel, Nsrsha, Pinot Meunier, Sangiovetto 포도주의 주요 향기성분은 과실 향인 것으로 나타났으며 Cabernet Franc, Cabernet Sauvignon, Sauvignon Vert 포도주의 주요 향기성분은 풀향인 것으로 조사되었다. 또한, MBA와 Narsha 포도주의 경우 꽃향이 주요 향인 것으로 조사되었다. 본 연구 결과를 바탕으로 적포도주용 품종을 육성할 때 선발기준으로 향기성분 분석을 활용할 수 있을 것으로 판단되었다.

Dendropanax morbifera Leveille (Araliaceae) is an endemic species growing in the south-western part of South Korea that has been used in folk medicine and health functional food. In this study, we investigated an extract of quercetin in Jeju D. morbifera by varying different parts (fruit, sprouts, leaves, sprigs, and branches), harvest times, and extraction solvents. In addition, we aimed to establish a simple and reliable HPLC/UV analytical method to determination of quercetin for the quality control and base line data of the Jeju D. morbifera extract as a health functional food ingredient. The analytical specificity was determined with retention time and photo diode array (PDA) spectrum by analyzing quercetin using HPLC and comparing the results to those of extracts. This analytical method for quercetin was validated for its limit of detection (LOD), limit of quantitation (LOQ), precision, and accuracy. A high linearity in the standard calibration curve was obtained, with a coefficient of determination (R2) of 0.9996. Also, the LOD and LOQ values were found to be 0.28 μg/mL and 0.85 μg/mL, respectively, and the recoveries of quantified compounds ranged from 97.91% to 104.10%. Furthermore, the relative standard deviation (RSD) values of data from the intra- and inter-day precision analyses were less than 1.36% and 3.65%, respectively. As a result, the highest quercetin content among the extracts of Jeju D. morbifera leaves was found to be 20.14 mg/g, which was extracted at harvest in May (cultivation period 10 years) with 60% EtOH. All in all, we believe that the results obtained would be helpful in the development of nutraceutics and natural medicines and for the quality control of D. morbifera.

Background : Heat stress induced from high temperature are known to crucially affecting on physiological properties and yield in Cnidium officinale. Methods and Results : The effect of foliar application of mixture including a urea, ascorbic acid and calcium chloride on high temperature injury of Cnidium officinale. Photosynthesis and leaf temperature in Cnidium officinale were investigated after cultivating for 24 hours at 35℃. Net photosyntheis rate, transpiration was measured at 1,000 μmol m-2 s-1 of photon flux density and leaf temperature was analyzed by thermal image. Net photosyntheis rate, stomatal conductance and transpiration rate in mixture traetment were 2 times of higher than in control. Water use efficience was not different significantly. Leaf temperature was lower in mixture treatment (25.3℃) than in control (29.0℃). Conclusion : This result show that foliar application of urea, ascorbic acid and calcium chloride was reducing a high temperature injury through a improving photosynthetical capacity and decreasing leaf temperature of Cnidium officinale.

Background : Management of air temperature are known to primarily affecting on physiological properties and yield in plant. Methods and Results : The effect of air temperature on characteristics of photosynthesis and chlorophyll fluorescence in Cnidium officinal were investigated using growth chamber after cultivating for 24 hours under controlled condition. Net photosyntheis rate, transpiration was measured at 1,000 μmol m-2 s-1 of photon flux density and chlorophyll fluorescence was analyzed by OJIP method. Net photosyntheis rate was highest in treatment of 25℃. Although transpiration rate was lowest, water use efficience was also in treatment of 25℃. Stomatal conductance was mainly influenced from ambient climatric factors such as vapor pressure deficit. As results of chlorophyll fluorescence by OJIP analysis, maximum quantum yield (Fv/Fm) of photosystem II (PSII), PIabs and the relative activities per reaction center such as ABS/RC, DIo/RC were not changed at air temperature. Therefore, elevated air temperatue during short term influence the dark reaction in photosystem through controlling a water use efficience and transpiration. Conclusion : This result show that 25℃ of air temperature may be a adequate temperature to improving the efficiency of photosynthesis in Cnidium officinale.

Background : Methicillin-Resistant Staphylococcus aureus (MRSA) is a multidrug-resistant (MDR) strain. Especially, MRSA is developing resistance to available antibacterial agents and causing complications in the treatment of infections related to skin, soft tissue, respiratory, bone, joint, and endovascular disorders. Therefore, antibacterial agent combination therapy appears to be a useful option, particularly in developing countries where antibiotic availability is limited. (+)-Usnic acid (UA) is uniquely found in lichens, and is especially abundant in genera such as Usnea and Cladonia. UA has antimicrobial activity against human and plant pathogens. Therefore, UA may be a good antibacterial drug candidate for clinical development. Methods and Results : In search of a natural products capable of inhibiting this multidrug-resistant bacteria, we have investigated the antimicrobial activity of UA against MRSA. In this study, the effects of a combination of UA and permeable agents against MRSA were investigated. For the measurement of cell wall permeability, UA with concentration of Ethylenediaminetetraacetic acid (EDTA) was used. In the other hand, Sodium azide (NaN3) was used as inhibitors of ATPase. These results suggest that the antibacterial effect of UA was potentiated by membrane-binding agents and ABC transporter-inhibiting agents, implying that antibacterial activity is associated with damage of the cell wall and inhibition of ATPase function by UA. Conclusion : UA and in combination with EDTA and NaN3 could lead to the development of new combination antibiotics against MRSA infection. The results of this study appear to be promising, and they are expected to enhance the use of natural products as drugs.

Background : Ginseng has been commonly used as a traditional oriental medicine for its wide spectrum of medicinal properties, including anti-inflammatory, antitumorigenic, adaptogenic and anti-aging properties. 20(S)-Protopanaxadiol (PPD), a intestinal metabolite of ginsenosides, is one of the active ingredients in ginseng. In this study, we have found inflammation-related genes regulated by 20(S)-PPD in mouse bone marrow-derived macrophage (BMDM) to elucidate the role of 20(S)-PPD in inflammatory signaling pathways. Methods and Results : We examined cell viability of BMDM cells after treatment of 20(S)-PPD and found that 20(S)-PPD has no cytotoxicity up to 10 uM. BMDM cells treated with none or 10uM 20(S)-PPD were used for RNA extraction and microarray analysis. It was found that 2 inflammation-related genes are upregulated and 4 genes are downregulated by 20(S)-PPD. Conclusion : These results can give clues to elucidate the role of 20(S)-PPD in inflammatory signaling pathways.

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 : An important feature of the nutrient solution is that they affect not only the growth but also quality of crops by changing nutrient uptake, especially due to changes of EC in nutrient solution. This study was carried out to investigate effect of EC in nutrient solution on growth and ginsenoside of ginseng. Methods and Results : EC in nutrient solution was controlled with 0.68, 0.84, 1.23, 1.41 dS/m. The root weight of ginseng treated by low EC levels in nutrient solution was higher during the initial of growth. However, the higher EC levels, the more increased the change rate of root weight from the initial to the middle of growth. The highest amount of ginsenoside was changed by growth period. Although the total amount of ginsenoside in root is highest treated by EC 0.68 dS/m at 45 days after treatment. the total amount of ginsenoside in root is highest treated by EC 1.23 dS/m at 135 days after treatment. Conclusions : EC in nutrient solution should to be controlled depending on the stage of growth and the part of use, i.e. root and leaves, when ginseng is cultivated through nutri-culture.

Background : Boron (B) is an essential element required for the growth of plant. It has a narrow range of optimal concentration from minimum to maximum thresholds than other micro-elements. The study was carried out to investigate to the influence of B excess concentrations on physiological disorder of leaf, growth and mineral concentration of ginseng to obtain basic information for physiological disorder diagnose. Methods and Results : The ginseng cultivar ‘Gumpoong’ was cultivated by hydroponic system for 2 months. The toxicity symptoms which ginseng leaves were curved downwardly and induced to chlorosis after beginning to dry the edge appeared on leaves more than 30 ppm compared to the control(3 ppm). The growth of ginseng was more decreased with higher B concentration. Mn uptake was also decreased as B concentration increased. It was found that B excess hindered the growth of ginseng and was reversely related to Mn uptake. Conclusions : More than B 30 ppm can negatively affect growth and mineral uptake. Consequently, B excess can occur physiological disorder of ginseng.

Background : For increasing saponin content of ginseng cultivated in shaded plastic house, this study was performed to investigate growth characteristics and saponin content of ginseng (Panax ginseng C.A. Mayer) according to foliar spray of germanium and water-soluble silicates processing. Methods and Results : Used a native species in this study is violet-stem variant that most commonly cultivated in ginseng’s farms. 1-year-old violet-stem variant was transplanted on March 24, 2015 and planting distance was 11 × 20 cm. Shading material of plastic house was used blue-white shading vinyl. The processing method of inorganic dissolved matter is as follows, we were diluted with germanium and water-soluble silicates to 500-fold, 1000-fold and investigated after foliar spray twice a month from May to September. The growth characteristics in above-ground part of 2-year-old ginseng was a good in the inorganic matters treatment compared to the control treatment, the difference of above-ground growth characteristics between the inorganic dissolved matters treatments was not significant. The growth characteristics in under-ground part of 2-year-old ginseng was a good in the inorganic matter treatments and root weight per plant in the 500-fold dilution of germanium was 12.4 g that increased by 29 % compared to the control (9.6 g). Crude saponin content of under-ground part was higher generally in inorganic matter treatments compared to the control (11.53 ㎎/g). In the 1000-fold dilution of water-soluble silicate and germanium (50:50), crude saponin content was the highest in 12.91 ㎎/g. Crude saponin content of above-ground part was higher generally in inorganic matter treatments compared to the control (61.76 ㎎/g). In the 1000-fold dilution of water-soluble silicate and germanium (50:50), crude saponin content was the highest in 65.69 ㎎/g. Conclusion : From the above results, we concluded that germanium and water-soluble silicates could be useful matters in promoting growth characteristics and saponin content of ginseng (Panax ginseng C.A. Mayer).

Background : This study was performed to set proper soil moisture tension for promoting seed emergence and yield of ginseng when direct seeding cultivation of ginseng was carried out in shaded plastic house. Methods and Results : The test cultivars were used Cheonpung, Yeonpung, Geumpung and seeds were sown on November 20, 2013. Irrigation starting point was set to 30, 40, 50 kPa and irrigation breakpoint was set to 20 kPa. Ginseng was cultivated in clay loam soil and shading material of plastic house were used blue-white shading vinyl. The emergence rate of 3-year-old ginseng cultivars in accordance with soil moisture tension indicated Cheonpung 97.0%, Yeonpung 95.0%, Geumpung 95.3% in each 50 kPa, 50 kPa, 30 kPa. In general, emergence rate of ginseng was higher tendency in 50 kPa (93.3∼97.0%). The average absolute soil moisture content during the growing season indicated a moisture content of 19.3% (30 kPa), 17.9% (40 kPa), 16.2% (50 kPa). Looking at the growth characteristics in above-ground part of 3-year-old ginseng cultivars according to soil moisture tension, Yeonpung, Cheonpung and Geumpung were good growth tendency in each 30kPa, 40kPa, 50kPa. On the other hand, looking at the growth characteristics in underground part of 3-year-old ginseng cultivars according to soil moisture tension, Cheonpung, Geumpung and Yeonpung were good growth tendency in each 30kPa, 40kPa, 50kPa. Yield per 10a was indicated that Yeonpung was the highest in the 50kPa to 456kg. Yield per 10a of Cheonpung and Geumpung indicated 347kg, 263kg in each 30kPa, 50kPa. Conclusion : From the above results, we concluded that ginseng(Panax ginseng C.A. Mayer) seems to be a difference of growth characteristics according to soil moisture tension. Therefore, it is need to manage soil moisture for each ginseng cultivars

Hepatocytes and hepatic progenitors derived from human ES cells may be a useful source for clinical application. Therefore, identification and purification of these cell types would be following important issues. There are very few candidate surface markers that can be used to identify and purify hepatic progenitor cells. In addition, indocyanine-green can be uptaken by mature hepatocytes, but cannot be applied for fluorescence activated cell sorting (FACS) due to its long emission wavelength. In the present study, we tested EpCAM as a potential marker for magnetic-activated cell sorting (MACS) of hepatic progenitors and also modified indocyanine-green into fluorescent indomonocarbocyanine for FACS-mediated sorting of mature hepatocytes after differentiation of human ES cells. Hepatic progenitor cells were sorted by MACS after incubation with anti-human EpCAM antibodies. After the final differentiation, the differentiated cells and mouse primary hepatocytes (control group) were incubated with indomonocarbocyanine and were sorted by FACS. MACS and immunocytochemistry data showed that approximately 45% of differentiated cells were EpCAM-positive cells. EpCAM-positive cells expressed α-fetoprotein, FOXa2, HnF4a, and CK18. Differentiation efficiency into albumin-positive cells was significantly higher in EpCAM-positive cells, compared to EpCAM-negative cells. Importantly, indomonocarbocyanine successfully stained cells that expressed ALB. Furthermore, FACS analysis data showed that the purity of hepatocytes that expressed albumin was significantly increased after purification of indomonocarbocyanine-positive cells. Our data demonstrated that human ES cell-derived hepatic progenitors can be efficiently isolated by MACS using EpCAM antibody. In addition, we also showed that indomonocarbocyanine can be successfully used to identify and purify mature hepatocytes using FACS.

Highly homogeneous and functional stem cell-derived hepatocyte-like cells (HLCs) are considered a promising option in the treatment of liver disease and the development of effective in vitro toxicity screening tool. However, the purity of cells and expression and/or activity of drug metabolizing enzymes in stem cell-derived HLCs are usually too low to be useful for clinical or in vitro applications. Here, we describe a highly optimized differentiation protocol, which produces more than 90% albumin-positive HLCs with no purification process. In addition, we show that hepatic enzyme gene expressions and activities were significantly improved by generating three-dimensional (3D) spheroidal aggregate of HLCs. The 3D differentiation method increased expressions of nuclear receptors that regulate the proper expression of key hepatic enzymes. Furthermore, a significantly increased hepatic functions such as albumin and urea secretion were observed in 3D hepatic spheroids and HLCs in the spheroid exhibited morphological and ultrastructural features of normal hepatocytes. Importantly, we show that repeated exposures to xenobiotics facilitated the functional maturation of HLC, as confirmed by increased expression of genes for drug metabolizing enzymes and transcription factors. In conclusion, the 3D culture system with repeated exposures to xenobiotics may be a new strategy for enhancing hepatic maturation of stem cell-derived HLCs as a cell source for in vitro high-throughput hepatotoxicity models.

Hepatocyte-like cells (HLCs) derived from human pluripotent stem cells have received extensive attention in the development of drug screening and toxicity testing. However, it has been reported that stem cell-derived HLCs showed hepatic functions that were too limited to be of use in drug screening and toxicity testing, possibly due to the lack of sufficient intercellular communication under conventional two-dimensional (2D) culture conditions. Therefore, a 3D differentiation system may overcome the in vitro limitation of 2D culture to produce stem cell-derived hepatocytes with mature metabolic functions. In this study, the feasibility of using a silicone-based spherofilm, specifically designed to produce spherical cell clusters, to generate uniformly sized 3D hepatic spheroids from hESCs was investigated. Hepatic spheroids generated on the spherofilm showed more homogenous size and shape than those generated in conventional low-attachment suspension culture dishes. Results of immunohistochemical analysis showed that expression of the mature hepatic marker albumin (ALB) increased over time during the hepatic maturation process. Furthermore, the 3D culture system mimicked the in vivo 3D microenvironment. Laminin, which is an important component of hepatic ECM, was expressed in hepatic spheroids. The results of immunohistochemical analysis indicated that the 3D culture environment is capable of generating an in vivo-like microenvironment. In addition, quantitative PCR analysis showed that the mature hepatic marker ALB and cytochrome P450 (CYP) enzymes CYP3A4 and CYP3A7 were expressed at higher levels in 3D culture than in 2D culture. This indicates that the 3D culture system is suitable for hepatic maturation and that our size-controlled 3D culture conditions might accelerate hepatic function. These results suggest that 3D hepatic spheroids significantly enhance metabolic maturation of hepatocytes derived from hESCs

To understand molecular mechanisms underlying adaptation of plant cells to saline stress and stress memory, we developed Arabidopsis callus suspension-cultured cells adapted to high salt. Adapted cells to high salt exhibited enhanced tolerance compared to control cells. Moreover, the salt tolerance of adapted cells was stably maintained even after the stress is relieved, indicating that the salt tolerance of adapted cells was memorized. Salt-adapted and stress memorized cells were densely aggregated and formed multi-layered cell lump. Cell morphology analysis using transmission electron microscopy indicated that cell wall thickness of salt-adapted cells was significantly induced compared to control cells. In order to characterize metabolic responses of plant cells during adaptation to high salt stress as well as stress memory, we compared metabolic profiles of salt-adapted and stress-memorized cells with control cells by using NMR spectroscopy. A principle component analysis showed clear metabolic discrimination among control, salt-adapted and stress-memorized cells. Compared with control cells, metabolites related to shikimate metabolism such as tyrosine, and flavonol glycosides, which are related to protective mechanism of plant against stresses were largely up-regulated in adapted cell lines. Moreover, coniferin, a precursor of lignin, was more abundant in salt-adapted cells than control cells. The results provide new insight into metabolic level mechanisms of plant adaptation to saline stress as well as stress memory.

Retroperitoneal liposarcoma is an infrequent malignancy in adulthood arising from mesenchymal tissues. It is often diagnosed when the tumor becomes too large, suppressing adjacent organs and causing symptoms. We report on a case of giant liposarcoma found in a 77-year-old female undergoing surgical resection.

MFG-E8 (Milk fat globule-epidermal growth factor VIII), also called lactadherin or BA46, SED1 is a glycoprotein found in milk and mammary epithelial cells, it is a major protein component associated with milk fat globule membrane. Previously, our study showed that expression of MFG-E8 is gradually increased with hepatic differentiation of human embryonic stem cells (hESCs). Therefore, we hypothesized that MFG-E8 would be an early cancer stem cell marker, which may predict cancer progression. Our results showed that MFG-E8 was expressed in various human cancer cell lines such as HepG2, Hep3B, and Huh7. Production and secretion of the MFG-E8 were also confirmed in the conditioned media of those three cell lines using enzyme-linked immunosorbent assay. Next, we analyzed the MFG-E8 expression in 11 clinical cases of cholangiocellular carcinoma (CC) and 33 cases of hepatocellular carcinoma (HCC) by immunohistochemistry and examined the potential correlation with β-catenin and AFP, which are known cancer markers. According to hitological criteria, the progression of HCC and CC was evaluated and classified into high, low, metastatic, and well-, moderate-, poor-differentiated, respectively. Statistical analysis indicated that incidence of both HCC and CC is significantly associated with male compared to female (P<0.05). Tumor size also has positive correlation with age (r2=08948). Our immunohistochemistry data showed that MFG-E8 was expressed both HCC and CC tissue. Interestingly, the MFG-E8 expression was significantly increased with cancer progression (P<0.05) in both cases. Additionally, b-cateninexpression was increased and its localization was changed from membrane to cytoplasm and nucleus with the degree of HCC. Likely b-catenin, AFP was also increased with the degree of HCC but it was not correlated with severalty of CC. Importantly, both AFP and b-catenin were highly co-localized with MFG-E8 in HCC. These results suggest that MFG-E8 may have important physiological roles and its expression in HCC and CC would be considered as an important prognostic factor.