Biological properties of antimicrobial peptides (AMPs) of hemimetabolous insect are poorly characterized in innate immunity field. To investigate the biochemical properties of hemimetabolous insect’s AMPs, we purified the pyrrhocoricin-like AMP from the hemolymph of Riptortus pedestris and then named as riptocin. We successfully determined the primary protein structure and its cDNA sequence. Interestingly, the determined cDNA revealed that riptocin precursor is composed of 12 repeating units of active riptocins, which implied that riptocin precursor might require to be processed to generate active riptocins by several unidentified processing enzymes. In order to characterize the bio-processing mechanisms of riptocin precursor, we generated the antibody against active riptocin. Using quantitative PCR and Western blot analyses, we showed that gene of riptocin was started to express from the fatbody after three hours post bacterial infection. To address our hypothesis that active riptocin is generated from riptocin precursor by several processing enzymes, we need to obtain the riptocin precursor. Currently, we are expressing the recombinant riptocin precursor using in vitro translation system. Meanwhile, we investigated whether naive hemolymph (naive HL), which may contain precursor riptocin, can generate active riptocin when riptocin precursor was co-incubation with bacteria-challenged hemolymph (active HL), which may contain all processing enzymes. Actually, when naive HL was incubated with active HL, antimicrobial activity was dramatically increased, suggesting that processing enzymes in active HL may induce processing of riptocin precursor to generate active riptocins.

The objective of this study was to determine the effect of macrophages on growth of human colon cancer cells. The results showed that co-culture of colon cancer cells with macrophages inhibited the growth of colon cancer cells (HCT116 and SW620) depending on the number of macrophages, RAW 264.7 cells, and activated THP-1 cells accompanied by down regulation of pSTAT3 in cancer cells. We also found that expression and release of cancer cell growth inhibitory cytokines, IL-1 receptor antagonist (IL-1ra) and IL-10, was increased in macrophages. Blocking of the STAT3 pathway with specific inhibitor and siRNA of STAT3 abolished the growth of colon cancer cells and expression of IL-1ra and IL-10. In addition, neutralization of IL-1ra and IL-10 with antibodies resulted in reversal of macrophage-induced inhibition of cancer cell growth. These data showed that IL-1ra and IL-10 released from macrophages inhibit growth of colon cancer cells through inhibition of the STAT3 pathway.

Porcine Circovirus Type2 (PCV2), a single-stranded DNA virus associated with Postweaning multisystemic wasting syndrome(PMWS) of swine, has two major open reading frames, ORF1 and ORF2. The genomic size and molecular weight of ORF2 is respectively 699bp, 28kDa. ORF2 encodes the capsid protein (structural protein) that has type-specific epitopes and is very immunogenic and associated with the induction of neutralizing antibodies, suggesting its potential use in diagnostic assays as well as vaccine development. For efficient production of the capsid proteins, we expressed the PCV2 ORF2 gene with baculovirus in the insect cells. In this study, PCV2 ORF2 was appropriately ligated into the baculovirus transfer vector, pBacPAK9 and pB9-Acpol19-110-EK. Sf21 cells were transfected with a mixture of the purified recombinant transfer vector and bAcGOZA. We generated and purified recombinant viruses containing PCV2 ORF2, and named rAc-B9-PCV2ORF2 and rAc-B9-19-110-EK-PCV2ORF2, respectively. Expression levels of capsid fusion proteins with a partial polyhedrin region of AcNPV more increased than recombinant proteins from non-fusion expressed. Also, expression efficiency increased over time and differed at MOI. As a results, fusion expression of porcine circovirus type2 ORF2 using baculovirus could be utilized as an alternative expression method to produce recombinant antigen against PCV2 infection and is worthy of further investigation.

Wrinkles are an outward sign of cutaneous aging appearing preferentially on ultraviolet B (UVB)-exposed areas. The anti-wrinkle effects of herbal extracts were investigated in an animal model. Female albino hairless mice (HR/ICR) were randomly allocated to the control group (non-irradiated vehicle), positive control group (UVB irradiated-vehicle), and two herbal extract mixture groups (HE-1 and HE-2). HE-1 included Glycyrrhizae radix, Rhei Rhizoma, Cornus officinalis, and Sesami semeni, and HE-2 included Swertia pseudo-chinensis, Sophora flavescens, Scutellaria baicalensis, and Salvia miltiorrhiza. The herbal extract mixtures were pre-treated dorsally with 0.2 ml per individual five times per week for four weeks prior to the start of UVB irradiation. At the fifth week, the animals were exposed to UVB irradiation for a subsequent eight weeks, three times per week. The intensity of irradiation showed a gradual increase, from 30 mJ/cm 2 to 240 mJ/cm2 (1 MED: 60 mJ/cm2 ). Dorsal skin samples were stained with H&E in order to examine the epidermal thickness. In addition, Masson-Trichrome staining was performed for determination of the amount of collagen fiber. Treatments with HE-1&2 resulted in an increase in the amount of collagen fiber, a better appearance, and fewer wrinkles, compared with the positive control. As determined by hydroxyproline assay, treatments with HE-1&2 led to a significant increase in the amount of collagen, compared with the positive control group (p<0.05). Chronic UVB irradiation to skin of hairless mice resulted in an increase in expression of matrix metalloproteinase-1 (MMP-1), however, treatments with HE-1&2 tended to decrease the expression of MMP-1. These results indicate that the herbal extracts used in this study have a preventive effect on UVB-induced wrinkle formation in a hairless mouse model, due in part to inhibition of MMP-1 expression and increment of collagen amount.

Iron catalyzes the production of free radicals, which can be related to a variety of pathological events, such as cancer and aging. The effect of dietary iron was investigated on formation of colonic aberrant crypt foci (ACF) induced by azoxymethane in male F344 rats. Animals were fed three different diets, including iron-deficient (3 ppm Fe), iron-normal (35 ppm Fe), and iron-overloaded (350 ppm Fe) diets for eight weeks. During the first and second weeks of the experiment, animals received two subcutaneous injections of azoxymethane (AOM, 15 mg/kg body weight) to induce ACF. After staining with methylene blue, the total numbers of ACF and aberrant crypts (AC) were counted on the colonic mucosa. Analysis of blood and serum was performed using a blood cell differential counter and an automatic serum analyzer. Iron-deficient diets induced a significant decrease in red blood cell counts and the values of hemoglobin concentration, hematocrit, mean corpuscular hemoglobin, and mean corpuscular volume, while an iron-overloaded diet did not affect these values. The iron-overloaded diet induced an increase in deposits of iron in the liver of rats, as determined by the ICP method and Perl’s staining. The numbers of ACF per colon showed a slight increase in iron-overloaded or iron-deficient rats, without statistical significance, compared to iron-normal rats. The number of total AC per colon in iron-overloaded rats was significantly higher than that in iron-normal rats (p<0.05). The number of large ACF (≥ 4 AC per ACF) in iron-overloaded rats was also significantly higher than that in iron-normal rats (p<0.05). These results suggest that dietary iron intake may play an important role in colon carcinogenesis in humans and animals.

Although hair disorders are not life threatening, a lot of people who suffer hair loss and/or hair thinning is increasing in accordance with changes in lifestyle and nutritional balance. The aim of this study was to examine the effects of herbal extracts on hair regrowth in C3H/HeJ mice. There were 6 experimental groups including distilled water (D.W.), 50% ethanol (a vehicle control), 3% minoxidil (a positive control), and 3 kinds of herbal extracts mixtures (C, D & E). The test compounds included followings; C : Glycyrrhizae radix, Rhei Rhizoma, Cornus officinalis and Sesami semeni, D : Viticus fructus, Pulsatilla chinensis, Gardenia fructus and Artemisiae argyi herb, E : Swertia pseudo-chinensis, Sophora flavescens Scutellaria baicalensis and Salvia miltiorrhiza. The animals were shaved with an electric clipper. The test compounds were daily treated to dorsal skin with 0.2 ml per mouse for 3 weeks. The topical application of the E test solution accelerated hair regrowth after 10 days faster than that of the positive and vehicle controls. The activities of alkaline phosphatase (ALP) and γ-glutamyl transpeptidase significantly were increased in all the treatment groups after 3 weeks, compared with D.W. group. Especially, the E test solution notably increased ALP activity compared with positive or vehicle control group. Epidermal growth factor (EGF) and vascular endothelial growth factor (VEGF) were increased in all the treatment groups after 3 weeks compared with D.W. group. These results suggest that the herbal extracts have hair regrowth effect by increasing enzyme activities and growth factors and it can be useful for treatment for alopecia in humans.

This study was carried out to investigate effect of herbal extracts mixtures on hair growth in an alopecia model of C57BL/6 mice. There were 4 experimental groups including distilled water (DW, negative control), 25% ethanol(EtOH, vehicle control) and two herbal extract mixtures (HE-1 and HE-2). HE-1 included Polygonum and Brassica campestris extracts and HE-2 included Mulberry root and Gardenia extracts. The 6 weeks-old C57BL/6 male mice were shaved with an electric clipper and the test compounds were topically treated with 0.2 ml per mouse daily for 3 weeks. The hair re-growth was photographically determined at days 0, 4, 7, 10, 14, 17, and 21. The number of mast cells which is an important modulator of hair growth was counted in 1 cm of dorsal skin section of mice. There were no clinical signs in all experimental groups. As the results of photometric analysis, the topical application of the herbal extracts (HE-1 & 2) for 2 weeks to dorsal skin accelerated hair re-growth remarkably faster than that of DW group (p＜0.05). Activity of alkaline phosphatase (ALP) was significantly increased in EtOH group compared to DW group (p＜0.05). Both herbal extract mixtures also increased the ALP activity, but it was not significantly different from DW. Treatment of mice with HE-2 significantly increased mast cell population compared to EtOH. Taken together, these results suggest that herbal extract mixtures used in this study may have hair-growth promoting activities and can be useful for treatment for male pattern baldness or alopecia in humans.

Background : This study was conducted to determine the optimal ginseng greenhouse shading material for minimizing heat injury and maximizing the yield of ginseng plants in an effort to develop a stable ginseng cultivation technology and contribute to popularizing ginseng greenhouse farming. Methods and Results : Three different types of greenhouse shading material were used: polyethylene (PE) film + black screen (95%), light scattering film + black screen (75%), and blue-white vinyl. Important experimental details were implemented in accordance with the Analysis Standard of Agricultural Test and Research set out by the Rural Development Administration. The average greenhouse temperature for the period of May through October was 20.0℃ under the PE film + black screen, 20.2℃under the light scattering film + black screen, and 19.7℃ under the blue/white vinyl. High average photosynthesis rates of ginseng plants were demonstrated by light scattering film+black screen (2.35 μmole CO2/㎡/s) and blue-white vinyl (2.38 μmole CO2/㎡/s), with the PE film + black screen showing a considerably lower photosynthesis rate (1.55 μmole CO2/㎡/s). Heat injury occurred in 0.3%, 10%, and 0.1% of the plants grown under the PE film + black screen, light scattering film + black screen, and blue-white vinyl, respectively. As for the roots, the blue-white vinyl outperformed the other shading materials in root length (25.7 ㎝), taproot length (5.9 ㎝), and the number of fine roots (23.2). The blue-hite vinyl also yielded the greatest root fresh weight (30.2 g), followed by the light scattering film + black screen (29.1 g), with the PE + black screen yielding by far inferior average weight (22.0 g). The highest crude saponin content was yielded by the light scattering film+black screen (16.61 ㎎/g), followed by blue-white vinyl (11.36 ㎎/g) and PE + black screen (11.0 ㎎/g). Conclusion : Comparing the effects of three different shading materials on greenhouse-cultivated 3-year-old ginseng plants, the blue/white vinyl outperformed two other shading materials in most of the growth characteristics, whereas the and the light scattering film + black screen yielded the highest ginsenoside content.

Background : For increasing saponin content of ginseng cultivated in shaded plastic house, this research was performed to investigate growth characteristics and saponin content of ginseng according to foliar spray of chitosan and water-soluble silicates processing. Methods and Results : The ginseng cultivars to be used this research were Cheonpung, Yeonpung and 1-year-old seedlings of Cheonpung, Yeonpung was transplanted on March 24, 2015. Planting density was 72 plant/3.3㎡and Shading material of plastic house used blue-white shading vinyl. The processing method of organic matter is as follows, we were applied chitosan (40 kg / 10a), silicate (3kg / 10a) to soil scheduled for ginseng cultivation with basal application. Then, we were diluted with chitosan and silicates to 1000-fold and investigated growth characteristics, saponin content of ginseng after foliar spray twice a month from May to September. Conclusion : Photosynthetic rate of ginseng according to treatment of organic matter was higher in the chitosan experimental group compared to the control group and photosynthetic rate of ginseng cultivars was the highest in Cheonpung’s silicate group(3.70 μmole CO2/㎡/s). The growth characteristics in above-ground part of 2-year-old ginseng was a good in the organic matter treatments compared to the control group. Above-ground part’s growth characteristics of ginseng according to treatment of organic matter showed a good trend in plant height, stem length, stem diameter of chitosan experimental group. The growth characteristics in under-ground part of 2-year-old ginseng was a good in the organic experimental groups. In Yeonpung of the chitosan experimental group, fresh root weight per plant was 9.2 g/plant that increased by 46 % compared to the control group. In Cheonpung of the silicate experimental group, fresh root weight per plant was 8.3 g/plant that increased by 56 % compared to the control group. In the chitosan experimental group, crude saponin content of under-ground part was Cheonpung(10.38 ㎎/g), Yeonpung(12.29 ㎎/g), control(11.53 ㎎/g), respectively. In the silicate experimental group, crude saponin content of under-ground part was Cheonpung(11.73 ㎎/g), Yeonpung(14.21 ㎎/g), respectively. Crude saponin content of ginseng according to treatment of organic matter was. generally higher in Yeonpung than Cheonpung.

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