Seon Woo Cha

Enhanced Immunomodulatory Activity of Gelatin-Encapsulated Rubus coreanus Miquel Nanoparticles

The aim of this work was to investigate the immunomodulatory activities of Rubus coreanus Miquel extract-loaded gelatin nanoparticles. The mean size of the produced nanoparticles was 143 ± 18 nm with a bandwidth of 76 nm in the size distribution and a maximum size of ~200 nm, which allows effective nanoparticle uptake by cells. Confocal imaging confirmed this, since the nanoparticles were internalized within 30 min and heterogeneously distributed throughout the cell. Zeta-potential measurements showed that from pH = 5 onwards, the nanoparticles were highly negatively charged, which prevents agglomeration to clusters by electrostatic repulsion. This was confirmed by TEM imaging, which showed a well dispersed colloidal solution. The encapsulation efficiency was nearly 60%, which is higher than for other components encapsulated in gelatin nanoparticles. Measurements of immune modulation in immune cells showed a significant effect by the crude extract, which was only topped by the nanoparticles containing the extract. Proliferation of B-, T- and NK cells was notably enhanced by Rubus coreanus-gelatin nanoparticles and in general ~2–3 times higher than control and on average ~2 times higher than ferulic acid. R. coreanus-gelatin nanoparticles induced cytokine secretion (IL-6 and TNF-α) from B- and T-cells on average at a ~2–3 times higher rate compared with the extract and ferulic acid. In vivo immunomodulatory activity in mice fed with R. coreanus-gelatin nanoparticles at 1 mL/g body weight showed a ~5 times higher antibody production compared to control, a ~1.3 times higher production compared to the extract only, and a ~1.6 times higher production compared to ferulic acid. Overall, our results suggest that gelatin nanoparticles represent an excellent transport vehicle for Rubus coreanus extract and extracts from other plants generally used in traditional Asian medicine. Such nanoparticles ensure a high local concentration that results in enhancement of immune cell activities, including proliferation, cytokine secretion, and antibody production.

Comparison of Growth Characteristics and Ginsenosides Content of 6-Year-Old Ginseng (Panax ginseng C. A. Meyer) by Drainage Class in Paddy Field

**Ginseng Research Division, Department of Herbal Crop Research, NIHHS, RDA, Eumseong 369-873, Korea.**Horticultural and Herbal Crop Environment Division, NIHHS, RDA, Suwon 440-706, Korea.ABSTRACT : To develop the practical cultivation for paddy field, we investigated the properties of paddy soil, growth char-acteristics and ginsenoside content of 6-year-old ginseng, Cheonpung variety between poor drainage class (PDC) and imper-fect drainage class (IDC). Groundwater level in PDC showed monthly small changes of 20~30㎝, while IDC showedmonthly great changes of 28~71 ㎝ depending on rainfall. Soil moisture content in PDC and IDC was 17.2%, 22.5%, respec-tively. Air temperature in IDC was lower than 0.3 ℃, while soil temperature was higher than 0.8 ℃ compare to PDC, respec-tively. Main soil color of PDC was grayish olive, while IDC was brownish olive. PDC showed yellowish mottles only atunderground of 20~40㎝, while IDC showed that at underground of 30~90㎝. IDC showed lower pH, EC, potassium, cal-cium and magnesium content, but higher organic matter, phosphate, and iron content than that of PDC, respectively. All ofEC, organic matter, potassium, calcium, and magnesium content were decreased, but iron content was increased at the sub-soil layers of PDC. All of EC, organic matter, phosphorus, and potassium content were decreased, but calcium and magne-sium content were increased at the subsoil layers of IDC. Root yield in IDC was more increased by 33% than that of PDC.The moisture content and rusty ratio of ginseng root in IDC were lower than that of PDC. Ginsenoside content in IDC washigher than that of PDC because the ratio of lateral and fine root showing relatively high content of ginsenoside was higherin IDC than that of PDC.Key Words : Panax ginseng, Poor Drainage Class, Imperfect Drainage Class, Paddy Soil, Root Yield, Ginsenoside

Enhancement of Cosmeceutical Activities of Berberis koreana Bark by High Pressure and Ultrasonification Extraction Processes

This study was performed to investigate the enhancement of cosmeceutical activities of Berberis koreana bark by different extraction processes. The extracts are WE (water extract at , control), USE (ultrasonification for 1 hours at with water), HPE (high pressure for 5 minutes at with water) and USE + HPE (ultrasonification process for 1 hours after high pressure for 5 minutes at with water), respectively. The cytotoxicity of the extracts was in the range of 24.02~26.94% at 1.0 mg/ml concentration. The USE + HPE showed the lowest cytotoxicity. Compared to the WE, total phenolic and flavonoid contents in the USE + HPE increased to 121.5% and 154.2%. The USE + HPE showed the highest activity at 1.0 mg/ml concentration in 1,1-diphenyl-2-picryl-hydrazyl (DPPH) radical scavenging, inhibition activity of xanthine oxidase and superoxide dismutase (SOD)-like test, respectively. Tyrosinase inhibition of WE, USE, HPE and USE + HPE at 1.0 mg/ml concentration was measured as 17.72, 19.62, 22.83 and 24.16%, respectively. Hyaluronidase inhibition activities of the USE + HPE were higher than 20.8%~29.5% of the WE. Our results suggested that the extracts from ultrasonification process after high pressure extraction has relatively high cosmeceutical activities, and that the bark of Berberis koreana could be considered as a candidate of new functional cosmetic agents.

Analysis of Mitochondrial DNA Sequence and Molecular Marker Development for Identification of Panax Species

This study describes the identification of Panax species using mitochondrial consensus primers. Initially, a total of thirty primers were tested in ten Korean ginseng cultivars and two foreign Panax species, P. quinquefolius and P. notoginseng. In the polymerase chain reaction (PCR) amplification results, three primers (cox1, nad1/2-3 and nad2/1-2) generated co-dominant polymorphic banding patterns discriminating Korean ginseng cultivars from P. quinquefolius and P. notoginseng. However, these primers could not generated polymorphisms among the Korean ginseng cultivars, and simply represented species-specific polymorphisms for P. quinquefolius and P. notoginseng. Primers PQ91 and PN418 were designed from the consensus sequence of nad1/2-3 region. Two banding patterns (A or B) were detected in PQ91. Korean ginseng cultivars and P. notoginseng shared the same banding pattern (A type) and P. quinquefolius was identified another banding pattern (B type). In the case of PN418, two banding patterns (A or B) were detected in the Korean ginseng cultivars and two foreign Panax species. Korean ginseng cultivars and P. quinquefolius shared the same banding pattern (A type) and P. notoginseng was identified another banding pattern (B type). The combination banding patterns of three Panax species, Korean ginseng cultivars (Panax ginseng C. A. Mey.), P. quinquefolius and P. notoginseng, was identified as `AA`, `BA` and `AB`, respectively. Consequently, PQ91 and PN418 primer sets can be used to distinguish among Panax species.

Cultivation Characteristics and Variation of Artemisinin Contents by Harvest Time of Artemisia annua L. Distributed in Korea

This study was conducted to obtain the basic data for using the Artemisia anuua as a new economic crop, thus Artemisia anuua was investigated their cultivation characteristics, yield, and variation of artemisinin contents by planting density and harvesting times. Seed characteristics of A. anuua have observed micro-size, and their germination optimum temperature was at 15 to 20 celsius degree. Planting density on the yield of A. anuua was increased high density better than low density. The highest yield was planted in the space of cm. Moreover, optimum harvesting time of A. anuua was investigated in early september and a periods of most highly detected artemisinin was time of before and after blooming of A. anuua.

Growth Characteristics and Ginsenoside Contents of 6-Year-Old Ginseng (Panax ginseng C. A. Meyer) by Shade Materials in Paddy Field

This study was carried out to select optimal shade materials among four-layered polyethylene (PE) net (FLPN), aluminium-coated PE sheet (APSS), and blue PE sheet (BPSS) in condition of paddy field cultivated 6-year-old gin- seng. The order of light-penetrated ratio and air temperature by shade materials was BPSS > APSS > FLPN. Light-pene- trated ratio of BPSS before two fold shade was more 3 times and 2 times than that of FLPN and APSS, respectively. Air temperature of BPSS was also higher 1.6 o C and 1.4 o C than that of FLPN and APSS, respectively. BPSS showed good cul- tural environment because all of light-penetrated ratio and air temperature were become higher in spring and fall season but lower in summer season by additional shade with two-layered PE net. Survived-leaf ratio was highest in BPSS and low- est in FLPN causing a little water leak on a rainy day. Rusty-root ratio was also highest in FLPN because soil moisture con- tent was increased by water leak. The order of root yield was BPSS > APSS > FLPN, and the cause of highest yield in BPSS was higher light-penetrated ratio during spring and fall season, higher survived-leaf ratio, and lower rusty-root ratio than that of APSS and FLPN. BPSS showed highest total ginsenoside content because of high light-penetrated ratio, blue light effect, and the difference in dry matter partitioning ratio such as low taproot ratio, and high lateral root ratio.

Effects of Different Germination Characteristics, Sowing Date and Rain Sheltered Cultivation on Stable Seed Production in Scutellaria baicalensis Georgi

Background: The purpose of this study was to evaluate methods to reduce seeding expenses, thereby increasing farm income. This study investigated the effects of rain shelter controlled cultivation and adapted seeding times on the stable seed production of Scutellaria baicalensis Georgi. Methods and Results: Seed germination was conducted under 10 condition compose to control, water washing, cold storage at for 15 days, seed sterilization with a benomyl pesticides, hormone treated seed by submerging in 100 ppm with the cold storage at and , darkness in a covered petridish and illuminated with a 1,500 Lux lamp. There were three cultivation type, open cultivation with non-woven fabric mulching, cultivation with a vinyl covering and rain sheltered in a plastic greenhouse. Sowing dates were April 27, May 18, June 7 and June 28, 2013. Plants were spaced 10 cm apart in rows 30 cm apart. Mixed oil cake fertilizer, (12-10-10) was applied at . Conclusions: Optimum germination occured in darkness at and cold storage after submerging in . The highest seed yields () occurred in the plastic greenhouse for the April 25 sowing. The highest root yield (17%) was found on April 1, under greenhouse conditions.

Clinical Study on Food Safety evaluation of Panax ginseng

*Department of Herbal Crop Research, NIHHS, RDA, Eumseong 369-873, Korea.**College of Chinese Medicinal Materials, Jilin Agricultural University, Changchun 130118, China.ABSTRACT : In this study, the clinical safety and toxicology of oral ingestion of supplement capsules containing ginsengradix was investigated in healthy young volunteers. This study was a pilot randomized, double blinded, placebo controlledtrial. The healthy volunteers were divided into 6 groups of 20 each (10 males and 10 females). They took the ginseng powderfor 35 days (3g/day) for safety evaluation. There were measured general healthy levels such as hematological, biochemical andelectrocardiographic parameters. After the first week, besides Korean white ginseng the other treatments led to an significantincrease of white blood cells. Korean red ginseng increased UREA (blood urea nitrogen) in healthy volunteers, but it didn’texceed the range of normal values, and in the subsequent process of treatment there is no effect of elevating UREA. After thethree weeks, Korean white ginseng showed relatively low the content of blood glucose and low-density lipoprotein cholesterol.After the five weeks, compared with the other treatments, Korean red ginseng increased white blood cells, platelet distributionwidth and average volume of platelet. Korean white ginseng decreased low-density lipoprotein cholesterol. American ginsengdecreased blood creatinine in healthy volunteers. In conclusion, through test the blood routine, urine routine, liver function,renal function, blood glucose, blood lipid and electrocardiogram, the healthy volunteers continuous taking ginsengfor 35 days (3 g/day) is safe and reliable, and have no obvious adverse reactions and side effects.Key Words : Panax ginseng, Panax quinquefolium, Food Safety

Overexpression of Farnesyl Diphosphate Synthase by Introducing CaFPS Gene in Panax ginseng C. A. Mey.

FPS (farnesyl diphosphate synthase) plays an essential role in organ development in plants. However, FPS has not previously been identified as a key regulatory enzyme in triterpene biosynthesis. In order to investigate the effect of FPS on ginsenosides biosynthesis, we over-expressed FPS of Centella asiatica (CaFPS) in Panax giseng adventitious roots. PCR analysis showed the integrations of the CaFPS and hygromycin phosphotransferase genes and we ultimately selected three lines. The result of Southern blot analysis demonstrated the introduction of the CaFPS gene into genome of ginseng. In addition, the results of RT-PCR analysis revealed that CaFPS gene overexpression induced an accumulation of its transcription in the ginseng adventitious roots. To determine whether or not the overexpression of the CaFPS gene contributes to the downstream gene expression associated with triterpene biosynthesis, the level of mRNAs was analyzed by real-time PCR. The result showed that no differences were detected in any expression of all genes. To determine quantitatively the content of ginsenosides in transgenic ginseng adventitious roots, HPLC analysis was conducted. The content of total 7 ginsenosides was increased to 1.8, 1.4, and 1.7 times than that of the controls, respectively. This indicated that the overexpression of CaFPS in ginseng adventitious roots causes an increase in ginsenoside content, although down stream genes of FPS gene were suppressed by CaFPS overexpression.

Variation of Yield and Loganin Content According to Harvesting Stage of Dipsacus asperoides Wall

Background: This study aimed to investigate the effect of harvest time on the growth, yield characteristics and loganin content in Dipsacus asperoides Wall. Methods and Results: Dipsacus asperoides seedlings were planted within a nursery environment in early May 2015 and harvested in early, middle and late October 2015, and early November 2015. Harvest time did not result significant differences in the plant height, stem diameter, branch length, leaf width and aboveground dry weight moreover, no significant differences were observed in root length, number of roots and root diameter. However, the diameter of lateral roots was greater in the harvests from the late October and period thereafter. The highest values of root dry weight and yield were recorded in early November. Specifically, the yield significantly increased from 205 kg/10 a (index: 100) in early October to 358 kg/10 a (index: 175) in early November, in terms of root part weight. Loganin contents of D. asperoides differed significantly among harvest times raging from 0.0766% in early October to 0.1704% in late November, thereby showing an increasing trend in later harvest times. Conclusions: These results suggest that the optimum harvest time for D. asperoides is early November, when the yield is the highest. Harvest time significantly affected loganin contents, which constantly increased from early October until early November.